ubiquinone and ubiquinone-9

Noise is a stress factor that causes auditory, psychological and physiological effects. The realization that sudden loud noises or chronic exposure to noise in social and working environments can cause hearing loss has led to increased interest in noise-induced hearing loss (NIHL). The best means of preventing primary damage is protection against noise. Since this protection is not always possible for various reasons, the use of pharmacological agents to prevent or treat NIHL should also be considered. The purpose of this study is to discuss current pharmacological protection and treatment options in the light of the literature, since no such extensive reviews have been performed to date, including agents used for protection against and treatment of NIHL. We reviewed both animal and clinical studies, and these are discussed separately for ease of comprehension. For each agent, first animal studies, then clinical studies, if available, are discussed. We also performed a two-step search of the literature. In the first step, we searched the terms "noise induced hearing loss", "treatment" and "protection" in Pubmed. Based on the results obtained, we identified the agents used for the treatment of and protection against NIHL. In the second step, we searched the names of the agents identified in the first step, together with the term "noise induced hearing loss," and reviewed the results.

Topics: Animals; Antioxidants; Calcium Channel Blockers; Dizocilpine Maleate; Excitatory Amino Acid Antagonists; Glucocorticoids; Hearing Loss, Noise-Induced; Humans; Magnesium; Methionine; Nerve Growth Factors; Ubiquinone; Vitamins

Topics: Animals; Chromatography, High Pressure Liquid; Coenzymes; Electrochemistry; Mitochondria, Liver; Ubiquinone; Vitamin E

Ubiquinone (coenzyme Q, Q) is an essential lipid electron carrier in the mitochondria respiratory chain, and also functions as antioxidant and participates as a cofactor of mitochondrial uncoupling proteins. Caernorhabditis elegans synthesize Q9, but both dietary Q8 intake and endogenous Q9 biosynthesis determine Q balance. Thus, it is of current interest to know the regulatory mechanisms of Q9 biosynthesis in this nematode. Here we review results that leaded to identification of genes involved in Q9 biosynthesis in this nematode using the RNA interference technology. C. elegans coq genes were silenced and depletion of Q content was observed, indicating that the genes related here participate in Q9 biosynthesis. Silenced populations showed an extension of adult life span, probably by the decrease of endogenous oxidative stress produced in mitochondria. We also report the heterologous complementation of C. elegans coq-5 and coq-7 genes in their homologue yeast coq null mutants, leading to restore its ability to growth in non-fermentable sugars. These complemented yeast strains accumulated Q6 but also the intermediate demethoxy-Q6. These findings support the conservative functional homology of these genes.

Topics: Animals; Caenorhabditis elegans; Genetic Complementation Test; Oxidative Stress; RNA Interference; Saccharomyces cerevisiae; Ubiquinone

Topics: Animals; Cholesterol, LDL; Combined Modality Therapy; Cytokines; Drug Evaluation, Preclinical; Fluid Therapy; Gene Expression Profiling; Humans; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Kaplan-Meier Estimate; Liver; Male; Mitochondria; Muscle, Skeletal; Myocardium; Oxygen Consumption; Rats, Wistar; Sepsis; Simvastatin; Tissue Culture Techniques; Ubiquinone

Between 82.8% and 92.5% of participants in any BMI group were responders by AS, and between 91.3% and 100% were responders by BBPS in the right colon. Efficacy was consistent across BMI groups, with no clear trends. Greater than 83% of participants in any BMI group found the preparation 'easy' or 'acceptable' to ingest, and the majority (>58%) rated SPMC oral solution as 'better' than a prior bowel preparation. In all BMI groups, safety data were similar to the overall cohort. Commonly reported, drug-related, treatment-emergent AEs were, by ascending BMI group, nausea (1.1%, 5.3%, 1.0%, 5.7%, and 0%) and headache (1.1%, 4.1%, 1.0%, 5.7%, and 0%).. Ready-to-drink SPMC oral solution had consistent, good quality colon cleansing, and favorable tolerability among participants of all BMI groups.. NCT03017235.. The pretreatment serum AST/ALT ratio predicts poor disease outcome and response rate in patients with advanced PDAC treated with gemcitabine/nab-paclitaxel and might represent a novel and inexpensive marker for individual risk assessment in the treatment of pancreatic cancer.. Of the 98 patients included in the study, 58 had CR (59%), 28 had PR (29%), and 12 patients had NR (12%). The percent splenic tissue embolized was significantly greater in the CR group compared to the PR group (P = 0.001). The percent volume of splenic tissue embolized was linearly correlated with the magnitude of platelet increase without a minimum threshold. At least one line of chemotherapy was successfully restarted in 97% of patients, and 41% of patients did not experience recurrence of thrombocytopenia for the duration of their survival. The major complication rate was 8%, with readmission following initial hospitalization for persistent "post-embolization syndrome" symptoms the most common.. In cancer patients with hypersplenism-related thrombocytopenia, PSAE is a safe intervention that effects a durable elevation in platelet counts across a range of malignancies and following the re-initiation of chemotherapy.. Postoperative CRP elevation was a better predictor of prognosis in patients with gastric cancer than the occurrence of intra-abdominal infectious complications.. In clinical practice, mixed-species malaria infections are often not detected by light microscopy (LM) or rapid diagnostic test, as a low number of parasites of one species may occur. Here, we report the case of an 8-year-old girl migrating with her family from Afghanistan with a two-species mixed infection with

Topics: 3-Hydroxybutyric Acid; Acetazolamide; Acrylates; Administration, Intravenous; Adolescent; Adult; Aerosols; Afghanistan; Aflatoxin M1; Agaricales; Aged; Aged, 80 and over; Agricultural Irrigation; Air Pollutants; alpha-L-Fucosidase; Amino Acid Sequence; Androgen Antagonists; Animals; Antibodies, Bacterial; Antigens, Bacterial; Antineoplastic Agents; Antioxidants; Apoptosis; Artifacts; Autophagy; B7-H1 Antigen; Bacterial Proteins; Bacterial Typing Techniques; Bariatric Surgery; Base Composition; Bayes Theorem; Bile; Bioelectric Energy Sources; Biosensing Techniques; Body Mass Index; Brain; Brazil; Breast Neoplasms; Bufo arenarum; Burkholderia; C-Reactive Protein; Cadmium; Carbon Compounds, Inorganic; Carbon-13 Magnetic Resonance Spectroscopy; Carbonic Anhydrase Inhibitors; Carbonic Anhydrases; Carcinoma, Transitional Cell; Case-Control Studies; CD4-Positive T-Lymphocytes; Cell Count; Cell Hypoxia; Cell Line, Tumor; Cell Proliferation; Characiformes; Child; China; Cities; Cobalt; Colonic Neoplasms; Copper Sulfate; Cross-Sectional Studies; Cyclin-Dependent Kinase Inhibitor p16; Cytokines; Deoxycytidine; Diagnosis, Differential; Digestive System; Dihydroxyphenylalanine; Disease Models, Animal; DNA (Cytosine-5-)-Methyltransferase 1; DNA Barcoding, Taxonomic; DNA, Bacterial; Dose-Response Relationship, Drug; Down-Regulation; Edetic Acid; Electrochemical Techniques; Electrodes; Embolization, Therapeutic; Embryo, Nonmammalian; Environmental Monitoring; Enzyme-Linked Immunosorbent Assay; Epithelial-Mesenchymal Transition; Fatty Acids; Feces; Female; Follow-Up Studies; Food Contamination; Forkhead Box Protein M1; Fresh Water; Fungicides, Industrial; Gallium Isotopes; Gallium Radioisotopes; Gastrectomy; Gastric Bypass; Gastric Outlet Obstruction; Gastroplasty; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Genes, Bacterial; Genetic Markers; Genome, Bacterial; Genome, Mitochondrial; Glioma; Glycogen Synthase Kinase 3 beta; Goats; Gonads; Guatemala; Halomonadaceae; HEK293 Cells; Helicobacter Infections; Helicobacter pylori; Hepacivirus; Histone-Lysine N-Methyltransferase; Hormones; Humans; Hydroxybutyrate Dehydrogenase; Hypersplenism; Hypoxia-Inducible Factor 1, alpha Subunit; Immunohistochemistry; Iran; Japan; Lactuca; Laparoscopy; Larva; Ligands; Liver Neoplasms; Lymphocyte Activation; Macrophages; Malaria; Male; Mercury; Metabolic Syndrome; Metals, Heavy; Mice; Middle Aged; Milk, Human; Mitochondria; Models, Molecular; Molecular Structure; Mothers; Multilocus Sequence Typing; Muscles; Mutation; Nanocomposites; Nanotubes, Carbon; Neoplasm Invasiveness; Neoplasm Recurrence, Local; Neoplasms; Neoplastic Cells, Circulating; Neoplastic Stem Cells; Neuroimaging; Nitriles; Nitrogen Isotopes; Non-alcoholic Fatty Liver Disease; Nuclear Magnetic Resonance, Biomolecular; Obesity; Obesity, Morbid; Oligopeptides; Oxidation-Reduction; Pancreatic Neoplasms; Particle Size; Particulate Matter; Pepsinogen A; Pesticides; Pharmacogenetics; Phosphatidylinositol 3-Kinases; Phospholipids; Phylogeny; Plasmodium ovale; Plasmodium vivax; Platelet Count; Polyhydroxyalkanoates; Positron Emission Tomography Computed Tomography; Positron-Emission Tomography; Postoperative Complications; Pregnancy; Prevalence; Prognosis; Prospective Studies; Prostate-Specific Antigen; Prostatic Neoplasms; Protein Domains; Proto-Oncogene Proteins c-akt; Proton Magnetic Resonance Spectroscopy; Pseudogenes; PTEN Phosphohydrolase; Pyrazoles; Pyrimidines; Radiographic Image Interpretation, Computer-Assisted; Radiopharmaceuticals; Rats, Long-Evans; Rats, Sprague-Dawley; RAW 264.7 Cells; Reactive Oxygen Species; Real-Time Polymerase Chain Reaction; Receptor, Notch3; Receptors, G-Protein-Coupled; Receptors, Urokinase Plasminogen Activator; Recombinant Proteins; Repressor Proteins; Resveratrol; Retrospective Studies; Risk Assessment; Risk Factors; RNA, Messenger; RNA, Ribosomal, 16S; Salinity; Salvage Therapy; Seasons; Sequence Analysis, DNA; Seroepidemiologic Studies; Signal Transduction; Skin; Snails; Soluble Guanylyl Cyclase; Solutions; Spain; Species Specificity; Spheroids, Cellular; Splenic Artery; Stomach Neoplasms; Streptococcus pneumoniae; Structure-Activity Relationship; Sulfonamides; Sunlight; Surface Properties; Surgical Instruments; Surgical Wound Infection; Survival Rate; Tetrahydrouridine; Thinness; Thrombocytopenia; Tissue Distribution; Titanium; Tomography, X-Ray Computed; TOR Serine-Threonine Kinases; Tumor Microenvironment; Tumor Necrosis Factor-alpha; Turkey; Ubiquinone; Urologic Neoplasms; Viral Envelope Proteins; Wastewater; Water Pollutants, Chemical; Weather; Wnt Signaling Pathway; Xenograft Model Antitumor Assays; Young Adult

Coenzyme Q (CoQ) is a redox-active lipid essential for core metabolic pathways and antioxidant defense. CoQ is synthesized upon the mitochondrial inner membrane by an ill-defined "complex Q" metabolon. Here, we present structure-function analyses of a lipid-, substrate-, and NADH-bound complex comprising two complex Q subunits: the hydroxylase COQ7 and the lipid-binding protein COQ9. We reveal that COQ7 adopts a ferritin-like fold with a hydrophobic channel whose substrate-binding capacity is enhanced by COQ9. Using molecular dynamics, we further show that two COQ7:COQ9 heterodimers form a curved tetramer that deforms the membrane, potentially opening a pathway for the CoQ intermediates to translocate from the bilayer to the proteins' lipid-binding sites. Two such tetramers assemble into a soluble octamer with a pseudo-bilayer of lipids captured within. Together, these observations indicate that COQ7 and COQ9 cooperate to access hydrophobic precursors within the membrane and coordinate subsequent synthesis steps toward producing CoQ.

Topics: Carrier Proteins; Humans; Lipids; Mitochondrial Membranes; Ubiquinone

Rheumatoid arthritis (RA) and its animal model adjuvant arthritis (AA) are inflammatory diseases characterized by chronic inflammation, systemic oxidative stress and disturbed mitochondrial bioenergetics of skeletal muscle. The present study aimed to evaluate the effects of coenzyme Q10 - CoQ10 (100 mg/kg b.w.), omega-3-polyunsaturated fatty acids - omega-3-PUFA (400 mg/kg b.w.) and their combined treatment in AA on impaired skeletal muscle mitochondrial bioenergetics, inflammation and changes in levels CoQ9 and CoQ10 in plasma. Markers of inflammation (C-reactive protein, monocyte-chemotactic protein-1), antioxidant capacity of plasma, respiratory chain parameters of skeletal muscle mitochondria and concentrations of CoQ9 and CoQ10 in plasma and in muscle tissue were estimated. Treatment of the arthritic rats with CoQ10, omega-3-PUFA alone and in combination partially reduced markers of inflammation and increased antioxidant capacity of plasma, significantly increased concentrations of coenzyme Q in mitochondria and improved mitochondrial function in the skeletal muscle. Combined treatment has similar effect on the mitochondrial function as monotherapies; however, it has affected inflammation and antioxidant status more intensively than monotherapies. Long-term supplementary administration of coenzyme Q10 and omega-3-PUFA and especially their combination is able to restore the impaired mitochondrial bioenergetics and antioxidant status in AA.

Topics: Animals; Antioxidants; Arthritis, Experimental; Arthritis, Rheumatoid; C-Reactive Protein; Chemokine CCL2; Dietary Supplements; Fatty Acids, Omega-3; Male; Mitochondria, Muscle; Rats, Inbred Lew; Ubiquinone

The genus

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Gammaproteobacteria; Nucleic Acid Hybridization; Phylogeny; Pigmentation; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone

Two Gram-staining-negative, aerobic, rod-shaped bacteria designated strains SR9

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Pseudomonas; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Soil Microbiology; Ubiquinone

During a study investigating the microbiota of raw milk and its semi-finished products, strains WS 5106

Topics: Animals; Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Genes, Bacterial; Germany; Milk; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Proteolysis; Pseudomonas; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone

Extracellular glutamine represents an important energy source for many cancer cells and its metabolism is intimately involved in maintaining redox homeostasis. The heightened metabolic activity within tumor tissues can result in glutamine deficiency, necessitating metabolic reprogramming responses. Here, dual mechanisms involving the stress-responsive transcription factor DDIT3 (DNA damage induced transcript 3) that establishes an interrelationship between glycolysis and mitochondrial respiration are revealed. DDIT3 is induced during glutamine deprivation to promote glycolysis and adenosine triphosphate production via suppression of the negative glycolytic regulator TIGAR. In concert, a proportion of the DDIT3 pool translocates to the mitochondria and suppresses oxidative phosphorylation through LONP1-mediated down-regulation of COQ9 and COX4. This in turn dampens the sustained levels of reactive oxygen species that follow glutamine withdrawal. Together these mechanisms constitute an adaptive survival mechanism permitting tumor cells to survive metabolic stress induced by glutamine starvation.

Topics: Animals; Apoptosis Regulatory Proteins; Electron Transport Complex IV; Energy Metabolism; Gene Expression Regulation, Neoplastic; Glutamine; Glycolysis; HCT116 Cells; Humans; Mice; Mice, Knockout; Neoplasms; Oxidative Phosphorylation; Phosphoric Monoester Hydrolases; Transcription Factor CHOP; Ubiquinone

Two

Topics: Animals; Bacterial Typing Techniques; Base Composition; Cattle; DNA, Bacterial; Fatty Acids; Genes, Bacterial; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Pseudomonas; Red Meat; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone

Background Coenzyme Q10 (CoQ10) serves as a shuttle for electrons from complexes I and II to complex III in the respiratory chain, and has important functions within the mitochondria. Primary CoQ10 deficiency is a mitochondrial disorder which has devastating effects, and which may be partially treated with exogenous CoQ10 supplementation. Case presentation A 9-month-old girl patient was referred to our clinic due to growth retardation, microcephaly and seizures. She was the third child of consanguineous parents (first-degree cousins) of Pakistani origin, born at 38 weeks gestation, weighing 2000 g after an uncomplicated pregnancy, and was hospitalized for 3 days due to respiratory distress. She had sustained clonic seizures when she was 4 months old. Physical examination showed microcephaly, truncal hypotonia and dysmorphic features. Metabolic tests were inconclusive. Abdominal ultrasonography revealed cystic appearance of the kidneys. Non-compaction of the left ventricle was detected in echocardiography. Cranial magnetic resonance imaging (MRI) showed hypoplasia of the cerebellar vermis and brain stem, corpus callosum agenesis, and cortical atrophy. A panel testing of 450 genes involved in inborn errors of metabolism (IEM) was performed that showed a novel frameshift c.384delG (Gly129Valfs*17) homozygous mutation in COQ9. A treatment of 5 mg/kg/day exogenous CoQ10 was started when she was 10 months old, and the dosage was increased to 50 mg/kg/day after the exact diagnosis. No objective neurological improvement could be observed after the adjustment of the drug dosage. Conclusions We report a case of CoQ10 deficiency due to a novel COQ9 gene mutation that adds clinical data from a newly diagnosed patient. Our case also outlines the importance of genetic panels used for specific diseases including IEM.

Topics: Ataxia; Female; Humans; Infant; Mitochondria; Mitochondrial Diseases; Muscle Weakness; Mutation; Prognosis; Rare Diseases; Ubiquinone

A novel bacterium, XHU 5135

Topics: Bacterial Typing Techniques; Base Composition; China; DNA, Bacterial; Fatty Acids; Halomonadaceae; Lakes; Nucleic Acid Hybridization; Phylogeny; RNA, Ribosomal, 16S; Salinity; Sequence Analysis, DNA; Ubiquinone

During investigations of spoilage-associated meat microbiota,

Topics: Animals; Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Food Microbiology; Genes, Bacterial; Germany; Meat; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Poultry; Pseudomonas; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Swine; Ubiquinone

Impaired mitochondrial function concomitant to enhanced oxidative stress-induced damage are well established mechanisms involved in hyperlipidemia-induced cardiotoxicity. Currently, limited information is available on the direct effect of myocardial lipid overload on endogenous coenzyme Q

Topics: Animals; Cell Line; Cell Respiration; Cell Survival; Mitochondria; Myocytes, Cardiac; Oxidative Stress; Palmitates; Rats; Reactive Oxygen Species; Ubiquinone

Ubiquinone, which is a component in the electron-transport systems of mitochondria, is essential for various activities related to energy metabolism, but the detailed absorption mechanism of ubiquinone is not clear. On the other hand, Niemann-Pick C1 Like 1 (NPC1L1) is involved in the intestinal absorption of fat-soluble components such as cholesterol. In this study, we investigated whether the intestinal absorption of ubiquinone was transported by NPC1L1 as is cholesterol. In this study, coenzyme q10 (CoQ10) and coenzyme q9 (CoQ9) were used as models of ubiquinone. The transport activity of ubiquinone was increased significantly in NPC1L1-overexpressed Madin-Darby canine kidney (MDCK) cells compared with that in pMAM2-BSD vector-transfected MDCK cells and the uptake of ubiquinone was decreased in the presence of ezetimibe, an inhibitor of NPC1L1. These results indicate that NPC1L1 mediates the transport of ubiquinone. Furthermore, to clarify the effect of NPC1L1 on the intestinal absorption of CoQ10, emulsified CoQ10 was orally administered to Wistar rats, and the plasma concentration was measured. The plasma concentration of CoQ10 was significantly decreased by coadministration of ezetimibe and CoQ10 compared to that with administration of only CoQ10. This result indicates that the intestinal absorption of CoQ10 is mediated by NPC1L1.

Topics: Administration, Oral; Animals; Dogs; Ezetimibe; Humans; Intestinal Absorption; Intestinal Mucosa; Madin Darby Canine Kidney Cells; Male; Membrane Transport Proteins; Micelles; Rats, Wistar; Ubiquinone

This study describes the biochemical and phylogenetic characteristics of a Gram-negative strain, SNU WT1

Topics: Animals; Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Genes, Bacterial; Kidney; Multilocus Sequence Typing; Oncorhynchus mykiss; Phosphatidylethanolamines; Phylogeny; Pseudomonas; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone

Seven endophytic strains were isolated from the halophyte Halimione portulacoides, collected from Ria de Aveiro, Portugal. To determine their exact taxonomic position, comparative analyses were performed with these strains and closely related type strains of Salinicola species. Genome sequencing and comparison indicated that five of the seven isolated strains comprised distinct and novel species (average nucleotide identity <0.95; in silico DNA-DNA hybridization <70 %; G+C difference >1 %). Multilocus sequence analysis was performed using gyrB, rpoD and 16S rRNA gene sequences from the novel and type strains to determine their phylogenetic positions. The novel strains are facultative anaerobes, mesophilic, facultative alkaliphic and halophilic, test positive for catalase and oxidase activities, for hydrolysis of Tween 20 and phosphate, for production of indole-3-acetic acid, but do not produce H2S. Ubiquinone UQ-9 is present in major amounts in all strains. The major fatty acids include C16 : 0 and the summed feature containing C18 : 1ω7c and/or C18 : 1ω6c. The DNA G+C content ranges from 60.6 to 65.8 mol%. Five strains were confirmed as new species belonging to the genus Salinicola, for which the names Salinicolahalimionae sp. nov. (type strain CPA60

Topics: Bacterial Typing Techniques; Base Composition; Chenopodiaceae; DNA, Bacterial; Fatty Acids; Genes, Bacterial; Halomonadaceae; Multilocus Sequence Typing; Nucleic Acid Hybridization; Phylogeny; Portugal; RNA, Ribosomal, 16S; Salt-Tolerant Plants; Sequence Analysis, DNA; Ubiquinone; Wetlands

Coenzyme Q (CoQ) deficiency has been associated with primary defects in the CoQ biosynthetic pathway or to secondary events. In some cases, the exogenous CoQ supplementation has limited efficacy. In the

Topics: Animals; Brain; Disease Models, Animal; Energy Metabolism; Histocytochemistry; Hydroxybenzoates; Mice; Mitochondrial Encephalomyopathies; Neuroprotective Agents; Salicylic Acid; Survival Analysis; Treatment Outcome; Ubiquinone

Coenzyme Q (CoQ) lipids are ancient electron carriers that, in eukaryotes, function in the mitochondrial respiratory chain. In mitochondria, CoQ lipids are built by an inner membrane-associated, multicomponent, biosynthetic pathway via successive steps of isoprenyl tail polymerization, 4-hydroxybenzoate head-to-tail attachment, and head modification, resulting in the production of CoQ. In yeast, we discovered that head-modifying CoQ pathway components selectively colocalize to multiple resolvable domains in vivo, representing supramolecular assemblies. In cells engineered with conditional ON or OFF CoQ pathways, domains were strictly correlated with CoQ production and substrate flux, respectively, indicating that CoQ lipid intermediates are required for domain formation. Mitochondrial CoQ domains were also observed in human cells, underscoring their conserved functional importance. CoQ domains within cells were highly enriched adjacent to ER-mitochondria contact sites. Together, our data suggest that CoQ domains function to facilitate substrate accessibility for processive and efficient CoQ production and distribution in cells.

Topics: Cell Line, Tumor; Endoplasmic Reticulum; Enzymes; Humans; Mitochondria; Mitochondrial Proteins; Multienzyme Complexes; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins; Substrate Specificity; Ubiquinone

The vast majority of mitochondrial disorders have limited the clinical management to palliative care. Rapamycin has emerged as a potential therapeutic drug for mitochondrial diseases since it has shown therapeutic benefits in a few mouse models of mitochondrial disorders. However, the underlying therapeutic mechanism is unclear, the minimal effective dose needs to be defined and whether this therapy can be generally used is unknown.. We have evaluated whether low and high doses of rapamycin administration may result in therapeutic effects in a mouse model (Coq9. Low dose of rapamycin induces metabolic changes in liver and transcriptomics modifications in midbrain. The high dose of rapamycin induces further changes in the transcriptomics profile in midbrain due to the general inhibition of mTORC1. However, neither low nor high dose of rapamycin were able to improve the mitochondrial bioenergetics, the brain injuries and the phenotypic characteristics of Coq9. These results may be due to the lack of microgliosis-derived neuroinflammation, the limitation to induce autophagy, or the need of a functional CoQ-junction. Therefore, the translation of rapamycin therapy into the clinic for patients with mitochondrial disorders requires, at least, the consideration of the particularities of each mitochondrial disease. FUND: Supported by the grants from "Fundación Isabel Gemio - Federación Española de Enfermedades Neuromusculares - Federación FEDER" (TSR-1), the NIH (P01HD080642) and the ERC (Stg-337327).

Topics: Animals; Autophagy; Cell Respiration; Disease Models, Animal; Gene Expression Profiling; Humans; Metabolomics; Mice; Mitochondria; Mitochondrial Diseases; Mitochondrial Encephalomyopathies; Phenotype; Sirolimus; Treatment Outcome; Ubiquinone

A Gram-stain-negative, aerobic, nitrogen-fixing bacterium, designated strain L461

Topics: Azotobacter; Bacterial Typing Techniques; Base Composition; China; DNA Fingerprinting; DNA, Bacterial; Fatty Acids; Kalanchoe; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Plant Leaves; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone

Primary CoQ

Topics: Acidosis, Lactic; Ataxia; Autopsy; Exome Sequencing; Female; Humans; Infant, Newborn; Leigh Disease; Male; Mitochondrial Diseases; Muscle Weakness; Mutation; Pregnancy; Siblings; Ubiquinone

Mitohormesis is an adaptive response induced by a mild mitochondrial stress that promotes longevity and metabolic health in different organisms. This mechanism has been proposed as the cause of the increase in the survival in Coq7

Topics: Animals; Antioxidants; Female; Longevity; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Mitochondria, Liver; Ubiquinone

A single missense mutation at position 159 of coenzyme Q9 (COQ9) (G→A; rs109301586) has been associated with genetic variation in fertility in Holstein cattle, with the A allele associated with higher fertility. COQ9 is involved in the synthesis of coenzyme COQ10, a component of the electron transport system of the mitochondria. Here we tested whether reproductive phenotype is associated with the mutation and evaluated functional consequences for cellular oxygen metabolism, body weight changes, and ovarian function. The mutation in COQ9 modifies predicted tertiary protein structure and affected mitochondrial respiration of peripheral blood mononuclear cells. The A allele was associated with low resting oxygen consumption and high electron transport system capacity. Phenotypic measurements for fertility were evaluated for up to five lactations in a population of 2273 Holstein cows. There were additive effects of the mutation (P < 0.05) in favor of the A allele for pregnancy rate, interval from calving to conception, and services per conception. There was no association of genotype with milk production or body weight changes postpartum. The mutation in COQ9 affected ovarian function; the A allele was associated with increased mitochondrial DNA copy number in oocytes, and there were overdominance effects for COQ9 expression in oocytes, follicle number, and antimullerian hormone concentrations. Overall, results show how a gene involved in mitochondrial function is associated with overall fertility, possibly in part by affecting oocyte quality.

Topics: Animals; Anti-Mullerian Hormone; Blastocyst; Body Weight; Cattle; Cell Respiration; Cumulus Cells; Endometrium; Energy Metabolism; Female; Fertility; Lactation; Mitochondria; Mutation, Missense; Oocytes; Ovary; Pregnancy; Ubiquinone

Within the frame of a biotechnological screening, we isolated two Pseudomonas strains from forest soil. 16S rRNA gene sequence analysis indicated that strain CCOS 864T shared 99.8 % similarity with Pseudomonas donghuensis HYST, while strain CCOS 865T shared 99.0 % similarity with Pseudomonas putida DSM 291T and lower similarity with other P. putida group type strains. Based on multilocus sequence analysis, the two strains were genotypically distinct from each other, each forming a separate clade. Strains CCOS 864T and CCOS 865T were Gram-stain-negative, motile and rod-shaped, growing at a temperature range of 4-37 °C. Strain CCOS 864T could be phenotypically distinguished from P. putida group species by the combination of gelatinase-positive reaction and positive growth on N-acetyl-d-glucosamine, p-hydroxyphenylacetic acid and inosine but lack of fluorescein production on King's B medium, while strain CCOS 865T could be distinguished from P. putida group species by the combination of positive growth with saccharic acid and negative growth with p-hydroxyphenylacetic acid and l-pyroglutamic acid. The major polar lipid for both strains was phosphatidylethanolamine; the major quinone was ubiquinone Q-9. DNA-DNA hybridization and average nucleotide identities confirmed the novel species status for the two strains. The DNA G+C contents of CCOS 864T and CCOS 865T were 62.1 and 63.8 mol%, respectively. The phenotypic, phylogenetic and DNA-DNA relatedness data support the suggestion that CCOS 864T and CCOS 865T represent two novel Pseudomonas species. The names Pseudomonas wadenswilerensis sp. nov. (type strain CCOS 864T=LMG 29327T) and Pseudomonas reidholzensis sp. nov. (type strain CCOS 865T=LMG 29328T) are proposed.

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Forests; Multilocus Sequence Typing; Nucleic Acid Hybridization; Phosphatidylethanolamines; Phylogeny; Pseudomonas; Pseudomonas putida; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Soil Microbiology; Switzerland; Ubiquinone

A novel Gram-stain-negative, rod-shaped and non-motile bacterial strain, designated strain Seoho-37T, was isolated from a eutrophic lake in South Korea. Polyphasic studies were performed to investigate the taxonomic position of the new isolate. The isolate grew aerobically with 0-1.0 % (w/v) NaCl (optimum 0 %), at pH 6.0-10.0 (optimum pH 7.0-9.0) and at temperatures of 15-36 °C (optimum 25-30 °C) on R2A medium. In the phylogenetic analysis of 16S rRNA gene sequences, strain Seoho-37T formed a clear cluster with the strains of Reyranella graminifolii, Reyranella massiliensis and Reyranella soli with a bootstrap resampling value of 100 %. DNA-DNA relatedness between strain Seoho-37T and the type strains of each species in the genus Reyranella was <20 %. The genomic DNA G+C content of strain Seoho-37T was 66.5 mol%. Ubiquinone-10 (Q-10) and ubiquinone-9 (Q-9) were found as the respiratory quinones. The cellular polar lipids were identified as diphosphatidylglycerol, phosphatidylglycerol and phosphatidylmethylethanolamine. The major fatty acid components included C16 : 0, summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c) and C18 : 1 2-OH. Based on the above evidence from a polyphasic study, strain Seaho-37T represents a novel species of the genus Reyranella, for which the name Reyranella aquatilis sp. nov. is proposed. The type strain is Seoho-37T (=KCTC 52223T=JCM 31892T).

Topics: Alphaproteobacteria; Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Eutrophication; Fatty Acids; Lakes; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Republic of Korea; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone

An endophytic bacterium, MA-69

Topics: Bacterial Typing Techniques; Base Composition; China; DNA, Bacterial; Fatty Acids; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Plant Stems; Populus; Pseudomonas; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone

Two Gram-stain-negative, aerobic, motile, halophilic, rod-shaped bacteria, designated Hb8T and Hb20, were isolated from a tidal flat environment located on the South-West Korean peninsula. The isolates grew at 10-37 °C, at pH 5.0-9.0 and in NaCl concentrations of 0.5-15 % (w/v; optimum, 3.0-6.0 %). Sequence analysis of the 16S rRNA indicated that the isolates belong to the genus Marinobacter and are most closely related to Marinobacter sediminumR65T (98.3 %), followed by Marinobacter lipolyticus SM19T, Marinobacter salsuginis SD-14BT and Marinobacter similis A3d10T. The overall 16S rRNA gene sequence similarity with these species was 97.9 %, but Hb8T and Hb20 showed 100 % sequence similarity with each other. DNA-DNA relatedness values of H8T and Hb20 suggested that these isolates represent a single species, while DNA-DNA relatedness values of the two novel isolates with M. sediminum DSM 27079T and M. similis DSM 15400T were only 21.3 and 22.9 %, respectively. The major fatty acids present in strain Hb8T were identified as C16 : 0, C16 : 1ω9c, C18 : 1ω9c, C18 : 0 3-OH and summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c). Ubiquinone-9 was the main respiratory quinone in both the novel strains. The polar lipids found to be present included diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, four unidentified phospholipids and five unidentified lipids. The genomic DNA G+C content of Hb8T and Hb20 was 54.5 mol%. Polyphasic analysis indicated that the two isolates are representatives of a novel species of the genus Marinobacte, for which the name Marinobacter salinus sp. nov. is proposed. The type strain is Hb8T (=KCTC 52255T=JCM 31416T).

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Marinobacter; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Republic of Korea; RNA, Ribosomal, 16S; Seawater; Sequence Analysis, DNA; Ubiquinone

A Gram-stain-negative, moderately halophilic, motile bacterium, designated strain CP12T, was isolated from a crystallizing pond of a saltern of the Yellow Sea in Korea. Cells of strain CP12T were non-spore-forming rods and produced whitish-yellow colonies. Growth was observed at 10-37 °C (optimum 37 °C), at pH 6.0-9.0 (optimum pH 7.0), and in the presence of 0.5-20 % (w/v) NaCl (optimum 3 %). Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain CP12T was closely related to Marinobacter flavimaris SW-145T (98.4 % 16S rRNA gene sequence similarity), Marinobacter algicola DG893T (98.2 %), Marinobacter adhaerens HP15T (98.2 %), Marinobacter salsuginis SD-14BT (97.9 %), Marinobacter salarius R9SW1T (97.6 %) and Marinobacter lipolyticus SM19T (97.1 %). DNA-DNA hybridization studies showed values lower than 18.6 % between strain CP12T and any of these species. The predominant respiratory isoprenoid quinone was ubiquinone-9 and the major cellular fatty acids of strain CP12T were C16 : 0, C12 : 0 3-OH, C12 : 0, Summed feature 3, C16 : 0 10-methyl and C18 : 1ω9c. On the basis of phenotypic properties, and phylogenetic and chemotaxonomic data, it is evident that strain CP12T represents a novel species of the genus Marinobacter, for which the name Marinobacter halotolerans sp. nov. is proposed. The type strain is CP12T (=KACC 18381T=NBRC 110910T).

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Marinobacter; Nucleic Acid Hybridization; Phylogeny; Ponds; Republic of Korea; RNA, Ribosomal, 16S; Salinity; Sequence Analysis, DNA; Ubiquinone

Coenzyme Q10 (CoQ10) has an important role in mitochondrial energy metabolism by way of its functioning as an electron carrier in the respiratory chain. Genetic defects disrupting the endogenous biosynthesis pathway of CoQ10 may lead to severe metabolic disorders with onset in early childhood. Using exome sequencing in a child with fatal neonatal lactic acidosis and encephalopathy, we identified a homozygous loss-of-function variant in COQ9. Functional studies in patient fibroblasts showed that the absence of the COQ9 protein was concomitant with a strong reduction of COQ7, leading to a significant accumulation of the substrate of COQ7, 6-demethoxy ubiquinone10. At the same time, the total amount of CoQ10 was severely reduced, which was reflected in a significant decrease of mitochondrial respiratory chain succinate-cytochrome c oxidoreductase (complex II/III) activity. Lentiviral expression of COQ9 restored all these parameters, confirming the causal role of the variant. Our report on the second COQ9 patient expands the clinical spectrum associated with COQ9 variants, indicating the importance of COQ9 already during prenatal development. Moreover, the rescue of cellular CoQ10 levels and respiratory chain complex activities by CoQ10 supplementation points to the importance of an early diagnosis and immediate treatment.

Topics: Acidosis, Lactic; Brain; Brain Diseases; Electron Transport Chain Complex Proteins; Fatal Outcome; Homozygote; Humans; Infant, Newborn; Male; Mitochondrial Proteins; Mutation; Ubiquinone; Ultrasonography

Strain THG-SQM11(T), a Gram-negative, aerobic, non-motile, coccus-shaped bacterium, was isolated from wheat seedlings plant in P. R. China. Strain THG-SQM11(T) was closely related to members of the genus Acinetobacter and showed the highest 16S rRNA sequence similarities with Acinetobacter junii (97.9 %) and Acinetobacter kookii (96.1 %). DNA-DNA hybridization showed 41.3 ± 2.4 % DNA reassociation with A. junii KCTC 12416(T). Chemotaxonomic data revealed that strain THG-SQM11(T) possesses ubiquinone-9 as the predominant respiratory quinone, C18:1 ω9c, summed feature 3 (C16:1 ω7c and/or C16:1 ω6c), and C16:0 as the major fatty acids. The major polar lipids were found to be diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, and phosphatidylcholine. The DNA G+C content was 41.7 mol %. These data, together with phenotypic characterization, suggest that the isolate represents a novel species, for which the name Acinetobacter plantarum sp. nov. is proposed, with THG-SQM11(T) as the type strain (=CCTCC AB 2015123(T) =KCTC 42611(T)).

Topics: Acinetobacter; Bacterial Typing Techniques; Base Composition; China; Fatty Acids; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Quinones; RNA, Ribosomal, 16S; Seedlings; Sequence Analysis, DNA; Species Specificity; Triticum; Ubiquinone

A moderately halophilic, aerobic bacterium, strain BZ-SZ-XJ27T, belonging to the genus Halomonas, was isolated from a saline-alkaline lake in the Xinjiang Uyghur Autonomous Region of China. Phylogenetic analysis based on 16S rRNA gene sequences and a multilocus sequence analysis using the 16S rRNA, gyrB and rpoD genes demonstrated that strain BZ-SZ-XJ27T represents a member of the genus Halomonas. On the basis of 16S rRNA gene sequence similarity, the closest relatives were Halomonas campaniensis 5AGT, H. fontilapidosi 5CRT, H. korlensis XK1T and H. sinaiensis ALO SharmT, with similarities of 96.2-97.2 %. DNA-DNA hybridization with H. korlensis CGMCC 1.6981T (the nearest phylogenetic neighbour) and H. campaniensis DSM 15293T (the highest 16S rRNA gene sequence similarity) showed relatedness values of 53 and 38 %, respectively, demonstrating the separateness of the three taxa. The bacterium stained Gram-negative and the cells were motile and rod-shaped. The strain formed creamy-white colonies and grew under optimal conditions of 1.42 M Na+ (range 0.22-4.32 M Na+), pH 8.0-8.5 (range pH 6.0-10.0) and 39 °C (range 4-43 °C). The dominant fatty acids were summed feature 8 (C18 : 1ω7c/C18 : 1ω6c; 36.6 %), C16 : 0 (25.9 %) and summed feature 3 (C16 : 1ω7c/C16 : 1ω6c; 21.2 %). The dominant polar lipids were two unknown phospholipids, phosphatidylethanolamine and phosphatidylglycerol, and the main respiratory quinones were ubiquinone 9 (Q-9; 89 %) and ubiquinone 8 (Q-8; 10 %). The genomic DNA G+C content was 61.7 ± 0.8 mol% (Tm). On the basis of phenotypic, chemotaxonomic and phylogenetic features, strain BZ-SZ-XJ27T is proposed to represent a novel species, Halomonas urumqiensis sp. nov., within the genus Halomonas of the family Halomonadaceae. The type strain is BZ-SZ-XJ27T ( = JCM 30202T = CGMCC 1.12917T).

Topics: Bacterial Typing Techniques; Base Composition; China; DNA, Bacterial; Fatty Acids; Genes, Bacterial; Halomonadaceae; Halomonas; Hydrogen-Ion Concentration; Lakes; Multilocus Sequence Typing; Nucleic Acid Hybridization; Phospholipids; Phylogeny; RNA, Ribosomal, 16S; Salinity; Sequence Analysis, DNA; Ubiquinone; Water Microbiology

Cephalotes 'turtle' ants are known to harbor a core group of gut symbionts, including members belonging to the Gammaproteobacteria. Here, we describe the cultivation and characterization of strain CV58T, a novel member of the Gammaproteobacteria order Pseudomonadales isolated from the guts of the ant Cephalotes varians. Strain CV58T was rod-shaped, Gram-stain-negative, non-motile and formed pale-yellow colonies on trypticase soy agar. Optimum growth occurred under an atmosphere of 4-20 % (v/v) O2. Growth was possible for strain CV58Tat NaCl concentrations of 0-1.5 % (w/v), temperatures of 23-40 °C, and pH values of 5.5-8.5. The G+C content of the genomic DNA was 54.9 mol% and the major fatty acids were C18 : 1ω7c, C16 : 0, C16 : 1ω7c/C16 : 1ω6c, C12 : 0 and C12 : 03OH. The only respiratory quinone detected was ubiquinone-9 (Q-9) and the major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. Based on phylogenetic analysis of the 16S rRNA gene sequence, strain CV58T shared an 88.3 % nucleotide identity with its closest cultivated neighbor, Pseudomonas putida R43. We believe that this, combined with the housekeeping gene phylogeny, differences in phenotypic characteristics and cellular fatty acid compositions of other cultivated members indicates that strain CV58T represents a novel species occupying a novel genus and family within the order Pseudomonadales. Thus, we propose the name Ventosimonadaceae fam nov., followed by Ventosimonas gracilis gen. nov., sp. nov., to classify strain CV58T (=NCIMB 15011T =DSM 100910T).

Topics: Animals; Ants; Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Gammaproteobacteria; Phospholipids; Phylogeny; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone

A moderately halophilic, Gram-stain-negative, non-endospore-forming endophytic bacterium designated strain ST307T was isolated from the euhalophyte Suaeda salsa in Dongying, China. Strain ST307T was aerobic, rod-shaped, motile and orange-yellow-pigmented. The organism grew at NaCl concentrations of 0.6-20 % (w/v) (optimum 5-6 %, w/v), at temperatures of 5-45 °C (optimum 35 °C) and at pH 5-9 (optimum pH 7-8). It accumulated poly-β-hydroxybutyric acid and produced exopolysaccharides. The major fatty acids were C18 : 1ω7c/C18 : 1ω6c, C16 : 0 and C16 : 1ω7c/C16 : 1ω6c. The predominant lipoquinone was ubiquinone Q-9. The polar lipids consisted of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, a glycoaminolipid and a phosphoglycoaminolipid. The DNA G+C content was 60.5 mol%. Phylogenetic analyses of 16S rRNA gene sequences and concatenated atpA, rpoD and secA gene sequences revealed that the strain represents a member of the genus Larsenimonas. The closest related type strain was Larsenimonas salina M1-18T. Mean DNA-DNA relatedness values between strain ST307T and the related species L. salina M1-18T, Chromohalobacter beijerinckii DSM 7218T, C. canadensis DSM 6769T, C. israelensis DSM 6768T, C. marismortui CGMCC 1.2321T, C. nigrandesensis DSM 14323T, C. salexigens DSM 3043T and C. sarecensis DSM 15547T were 15±2-45±1 %. On the basis of phenotypic, chemotaxonomic and molecular features, strain ST307T clearly represents a novel species of the genus Larsenimonas. The name Larsenimonassuaedae sp. nov. is proposed, with ST307T (=CGMCC 1.8902T=DSM 22428T) as the type strain.

Topics: Bacterial Typing Techniques; Base Composition; Chenopodiaceae; China; DNA, Bacterial; Fatty Acids; Halomonadaceae; Hydroxybutyrates; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Pigmentation; Polyesters; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone

The Coq protein complex assembled from several Coq proteins is critical for coenzyme Q6 (CoQ6) biosynthesis in yeast. Secondary CoQ10 deficiency is associated with mitochondrial DNA (mtDNA) mutations in patients. We previously demonstrated that carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP) suppressed CoQ10 levels and COQ5 protein maturation in human 143B cells.. This study explored the putative COQ protein complex in human cells through two-dimensional blue native-polyacrylamide gel electrophoresis and Western blotting to investigate its status in 143B cells after FCCP treatment and in cybrids harboring the mtDNA mutation that caused myoclonic epilepsy with ragged-red fibers (MERRF) syndrome. Ubiquinol-10 and ubiquinone-10 levels were detected by high-performance liquid chromatography. Mitochondrial energy status, mRNA levels of various PDSS and COQ genes, and protein levels of COQ5 and COQ9 in cybrids were examined.. A high-molecular-weight protein complex containing COQ5, but not COQ9, in the mitochondria was identified and its level was suppressed by FCCP and in cybrids with MERRF mutation. That was associated with decreased mitochondrial membrane potential and mitochondrial ATP production. Total CoQ10 levels were decreased under both conditions, but the ubiquinol-10:ubiquinone-10 ratio was increased in mutant cybrids. The expression of COQ5 was increased but COQ5 protein maturation was suppressed in the mutant cybrids.. A novel COQ5-containing protein complex was discovered in human cells. Its destabilization was associated with reduced CoQ10 levels and mitochondrial energy deficiency in human cells treated with FCCP or exhibiting MERRF mutation.. The findings elucidate a possible mechanism for mitochondrial dysfunction-induced CoQ10 deficiency in human cells.

Topics: Ataxia; Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; Cell Line; DNA, Mitochondrial; Humans; Membrane Potential, Mitochondrial; MERRF Syndrome; Methyltransferases; Mitochondria; Mitochondrial Diseases; Mitochondrial Proteins; Muscle Weakness; Mutation; RNA, Messenger; Ubiquinone

Eight Gram-stain-negative bacteria (B4199T, C6819, C6918, D2441, D3318, E1086, E1148 and E5571) were identified during a retrospective study of unidentified strains from a historical collection held in the Special Bacteriology Reference Laboratory at the Centers for Disease Control and Prevention. The strains were isolated from eight patients: five female, two male and one not specified. No ages were indicated for the patients. The sources were urine (3), leg tissue (2), foot wound, lung tissue and deep liver. The strains originated from seven different states across the USA [Colorado, Connecticut (2), Indiana, North Carolina, Oregon and Pennsylvania]. The strains grew at 10-42 °C, were non-motile, alkalitolerant, slightly halophilic, microaerophilic, and catalase- and oxidase-positive. The DNA G+C content was 47.3-47.6 mol%. The major cellular fatty acids were tetradecanoic acid (C14 : 0), hexadecanoic acid (C16 : 0) and 11-octadecenoic acid (C18 : 1ω7c). Polar lipids detected were phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol and unknown phospholipids; the only respiratory quinone detected was the ubiquinone Q-9 (100 %). 16S rRNA gene sequence analysis produced results with 95.6 % similarity to Pseudomonas caeni DSM 24390T and 95.2 % similarity to Thiopseudomonas denitrificans X2T. The results of the biochemical, chemotaxonomic and phylogenetic analyses between the study strains and some related type strains indicated that these strains represent a novel species of a new genus within the family Pseudomonadaceae, for which the name Oblitimonas alkaliphila gen. nov., sp. nov. is proposed. The type strain is B4199T (=DSM 100830T=CCUG 67636T).

Topics: Bacterial Typing Techniques; Base Composition; Colorado; Connecticut; DNA, Bacterial; Fatty Acids; Humans; Indiana; North Carolina; Oregon; Pennsylvania; Phospholipids; Phylogeny; Pseudomonadaceae; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone

A short-rod-shaped moderately halophilic bacterium, designated CUG 00002(T), was isolated from the sediment of Xiaochaidan salt lake in Qinghai Province, China by using R2A medium. The cells were Gram-staining negative, aerobic, forming creamy and circular colonies with diameters of 2-3 mm on R2A agar when incubated at 30 °C for 3 days. 16S rRNA gene-based phylogenetic analysis indicated that strain CUG 00002(T) belonged to the genus Halomonas in the class Gammaproteobacteria, showing highest sequence similarity of 97.1 and 96.7 % to Halomonas mongoliensis Z-7009(T) (=DSM 17332=VKM B2353) and Halomonas shengliensis SL014B-85(T) (=CGMCC 1.6444(T)=LMG 23897(T)), respectively. The predominant isoprenoid quinone was ubiquinone-9 (Q9), and the major fatty acids were C16:0, summed feature 3 (comprising C16:1 ω7c and/or C16:1 ω6c) and summed feature 8 (comprising C18:1 ω7c or C18:1 ω6c). The genomic DNA G+C content of strain CUG 00002(T) was 61.8 mol%. The above characteristics were consistent with the placement of the organism in the genus Halomonas. The level of DNA-DNA relatedness between CUG 00002(T) and its most closely related strain H. mongoliensis Z-7009(T) was 41.0 ± 1.6 %. Based on the results of phenotypic, phylogenetic and biochemical analyses, strain CUG 00002(T) represents a novel species of the genus Halomonas, for which the name Halomonas xiaochaidanensis sp. nov. is proposed. The type strain is CUG 00002(T) (=CCTCC AB 2014152(T)=KCTC 42685(T)).

Topics: Bacterial Typing Techniques; Base Composition; China; DNA, Bacterial; DNA, Ribosomal; Fatty Acids; Geologic Sediments; Halomonas; Hydrogen-Ion Concentration; Lakes; Nucleic Acid Hybridization; Phylogeny; Quinones; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Sodium Chloride; Soil Microbiology; Tibet; Ubiquinone

A Gram-stain-negative, non-endospore-forming, strictly aerobic, irregular rod-shaped bacterium without flagellum, designated strain H94T, was isolated by the high-throughput cultivation method from seawater of an amphioxus breeding zone in the coastal region of Qingdao, China. Growth was observed at 4-37 °C (optimum 28 °C), at pH 6.0-10.0 (optimum pH 7.0) and in the presence of 1-12 % (w/v) NaCl (optimum 1-2 %). The predominant cellular fatty acids were C18 : 1ω9c, C16 : 0 and C16 : 1ω9c. The polar lipids comprised phosphatidylethanolamine, phosphatidylglycerol and an unidentified phosphoglycolipid. The major respiratory quinone was ubiquinone-9 (Q-9). The genomic DNA G+C content of strain H94T was 56.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain H94T shared the highest similarity (95.9 %) with Hahella antarctica NBRC 102683T, and exhibited 92.9 % and 92.1 % similarity with the two other recognized Hahella species, Hahella chejuensis KCTC 2396T and Hahella ganghwensis DSM 17046T, respectively. The phylogenetic position revealed that strain H94T formed a stable distinct lineage cluster together with Hahella antarctica NBRC 102683T and this result was further confirmed by multilocus sequence analysis based on housekeeping genes gyrB and rpoB. On the basis of the polyphasic taxonomic analyses, strain H94T is considered to represent a novel species in a new genus, for which the name Allohahellamarinimesophila gen. nov., sp. nov. is proposed. The type strain of Allohahellamarinimesophila is H94T (=CGMCC 1.10800T=JCM 17555T). It is also proposed that Hahella antarctica should be reclassified within the genus Allohahella as Allohahella antarctica comb. nov. (type strain NBRC 102683T=IMCC 3113T=KCCM 42675T). The type species of the genus Allohahella is Allohahella antarctica comb. nov.

Topics: Antarctic Regions; Bacterial Typing Techniques; Base Composition; China; DNA, Bacterial; Fatty Acids; Gammaproteobacteria; Phospholipids; Phylogeny; RNA, Ribosomal, 16S; Seawater; Sequence Analysis, DNA; Ubiquinone

A Gram-stain-negative, moderately halophilic bacterium, designated strain CPS11T, was isolated from the sediment of a solar pond located in Shinan, Korea. Strain CPS11T was a strictly aerobic, motile, straight rod-shaped bacterium that grew at pH 5.0-9.0 (optimum, pH 7.0-8.0), at 10-37 °C (optimum, 28 °C) and at salinities of 1-20 % (w/v) NaCl (optimum, 10 % NaCl). Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain CPS11T belonged to the genus Halomonas, with sequence similarity of 98.5-94.3 % to existing type strains, showing highest sequence similarity to Halomonasfontilapidosi 5CRT (98.5 %), Halomonasventosae Al12T (98.5 %), Halomonascampaniensis 5AGT (98.2 %), Halomonas huangheensis BJGMM-B45T (98.0 %), Halomonas alimentaria YKJ-16T (98.0 %), Halomonas mongoliensis Z-7009T (97.8 %), Halomonas shengliensis SL014B-85T (97.5 %) and Halomonas cupida DSM 4740T (97.5 %). The predominant ubiquinone was Q-9. The major fatty acids were C19 : 0 cyclo ω8c, C16 : 1ω7c and/or iso-C15 : 0 2-OH, C16 : 0, C17 : 0 cyclo, C12 : 0 3-OH and C18 : 1ω7c. The polar lipids were phosphatidylglycerol, phosphatidylethanolamine, aminophospholipid, an unknown phospholipid and unknown lipids. The DNA G+C content of this novel isolate was 64.3 mol%. Levels of DNA-DNA relatedness between strain CPS11T and the type strains of ten other species of the genus ranged from 50 to 21 %. On the basis of the polyphasic analysis conducted in this study, strain CPS11T represents a novel species of the genus Halomonas, for which the name Halomonas sediminicola sp. nov. is proposed. The type strain is CPS11T(=KACC 18262T=NBRC 110636T).

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Geologic Sediments; Halomonas; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Ponds; Republic of Korea; RNA, Ribosomal, 16S; Salinity; Sequence Analysis, DNA; Ubiquinone

A Gram-negative, aerobic, non-motile bacterium, designated strain KC90BT, was isolated from the surface of a cell of the marine diatom Thalassiosira delicatula. The bacterial cells were pleomorphic and formed very small, beige colonies on marine agar. Optimal growth was obtained at 25 °C, at pH 6.5-7.5 and in the presence of 1.5-2.0 % (w/v) NaCl. Phylogenetic analyses based on its 16S rRNA gene sequence revealed that strain KC90BT belonged to the Roseobacter clade and formed a monophyletic cluster with the sequences of Boseongicola aestuarii, Profundibacterium mesophilum, Hwanghaeicola aestuarii, Maribius pelagius and M. salinus, showing 91.4-95.7 % sequence similarities. Ubiquinone Q-10 was the predominant lipoquinone but a significant amount of ubiquinone Q-9 was also detected. The major cellular fatty acids were C18 : 1ω7c, 11-methyl C18 : 1ω7c and C18 : 0. Strain KC90BT also contained specific fatty acids (C17 : 0, anteiso-C15 : 0 and anteiso-C17 : 0) that were not detected in its closest described relatives. The major polar lipids of strain KC90BT comprised phosphatidylglycerol, phosphatidylcholine, diphosphatidylglycerol and an unidentified aminolipid. The DNA G+C content of strain KC90BT was 65.2 mol%. The phylogenetic analysis of strain KC90BT, together with the differential phenotypic and chemotaxonomic properties demonstrate that strain KC90BT is distinct from type strains of B. aestuarii, P. mesophilum, H. aestuarii, M. pelagius and M. salinus. Based on the data presented in this study, strain KC90BT represents a novel genus and species within the family Rhodobacteraceae, for which the name Silicimonas algicola gen. nov., sp. nov. is proposed. The type strain is KC90BT (=DSM 103371T=RCC 4681T).

Topics: Bacterial Typing Techniques; Base Composition; Diatoms; DNA, Bacterial; Fatty Acids; Phospholipids; Phylogeny; Rhodobacteraceae; RNA, Ribosomal, 16S; Seawater; Sequence Analysis, DNA; Ubiquinone

A bacterial strain named IB1.1T was isolated in a screening of hydrocarbon-degrading bacteria from oil-contaminated soils on the territory of the Turukhansk District of Krasnoyarsk Krai, East Siberia, Russia. The 16S rRNA gene sequence had 98.7 % identity with respect to the closest phylogenetic relative, Pseudomonas granadensis F-278,770T, and the next most closely related species with 98.6 % similarity was Pseudomonaspunonensis, suggesting that IB1.1T should be classified within the genus Pseudomonas. The analysis of housekeeping genes rpoB, rpoD and gyrB showed similarities lower than 90 % in all cases with respect to the closest relatives, confirming its phylogenetic affiliation. The strain showed a polar flagellum. The respiratory quinone was Q9. The major fatty acids were 16 : 1ω7c/16 : 1ω6c (summed feature 3), 18 : 1ω7c and 16 : 0. The strain was oxidase- and catalase-positive, but the arginine dihydrolase system was not present. Nitrate reduction, urease and β-galactosidase production, and aesculin hydrolysis were negative. The temperature range for growth was 4-34 °C, and the strain could grow at pH 11. The DNA G+C content was 58.5 mol%. DNA-DNA hybridization results showed values of less than 30 % relatedness with respect to the type strains of the eight most closely related species. Therefore, the dataset of genotypic, phenotypic and chemotaxonomic data support the classification of strain IB1.1T into a novel species of the genus Pseudomonas, for which the name Pseudomonasturukhanskensis sp. nov. is proposed. The type strain is IB1.1T (=VKM B-2935T=CECT 9091T).

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Nucleic Acid Hybridization; Petroleum Pollution; Phylogeny; Pseudomonas; RNA, Ribosomal, 16S; Russia; Sequence Analysis, DNA; Siberia; Soil Microbiology; Ubiquinone

A novel, Gram-stain-negative, facultatively anaerobic, halophilic bacterium, designated strain Q1UT, was isolated from a sediment sample collected from Qinghai Lake, PR China. The cells of the strain were short rod-shaped (0.2-0.3×0.6-2.5 µm) and non-motile. Strain Q1UT formed yellowish colonies and grew at temperatures of 2-37 °C (optimum 30-33 °C), at pH 6.0-9.0 (optimum pH 7.0) and in the presence of 0-20 % (w/v) NaCl (optimum 7.5 %). The major cellular fatty acids were C18 : 1ω7c (58.6 %), C16 : 1ω7c and/or C16 : 1ω6c (14.8 %) and C16 : 0 (10.1 %). The polar lipids were identified as diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, unknown phospholipid and unknown lipids. The genomic DNA G+C content was 61.5 mol%, and the predominant respiratory ubiquinone Q-9. Based on phylogenetic analysis of the 16S rRNA gene sequences and concatenated 16S rRNA, gyrB and rpoD gene sequences, the isolate was found to belong to the genus Halomonas in the class Gammaproteobacteria. The most closely related species were Halomonas venusta DSM 4743T (98.3 % 16S rRNA sequence similarity), Halomonas songnenensis DSM 25870T (98.2 %) and Halomonas hydrothermalis DSM 15725T (98.2 %). DNA-DNA relatedness values between strain Q1UT and the type strains of eight other species of the genus Halomonas ranged from 21.3 % to 10.1 %. On the basis of phenotypic, phylogenetic and chemotaxonomic analyses, and DNA-DNA hybridization relatedness values, strain Q1UT is considered to represent a novel species of the genus Halomonas; the name Halomonas lutescens sp. nov. is proposed. The type strain is Q1UT (=CGMCC 1.15122T=KCTC 42517T).

Topics: Bacterial Typing Techniques; Base Composition; China; DNA, Bacterial; Fatty Acids; Geologic Sediments; Halomonas; Lakes; Nucleic Acid Hybridization; Phospholipids; Phylogeny; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone

A Gram-reaction-negative, aerobic, rod-shaped and non-motile marine bacterium, designated MEBiC09566(T) was isolated from a sponge collected at Uljin County in the coastal area of the East Sea (36° 55' N, 129° 25' E), Korea. The 16S rRNA gene sequence analysis revealed that strain MEBiC09566(T) showed the highest similarity with the Kiloniella laminariae LD81(T) (96.7%). Growth was observed at 11-31 °C (optimum 25 °C), at pH 6.0-8.5 (optimum pH 7.0) and with 0-6% (optimum 2.5%) NaCl. The predominant cellular fatty acids were summed feature 8 (comprised of C18:1ω7c/C18:1ω6c) and summed feature 3 (comprised of C16:1ω7c and/or C16:1ω6c). The DNA G+C content is 44.6 mol%. The major respiratory quinone is Q-9. Phosphatidylethanolamine, phosphatidylglycerol, an unidentified lipid, two unidentified aminophospholipids and one unidentified aminolipid were detected as major polar lipids. On the basis of this polyphasic taxonomic data, it is concluded that strain MEBiC09566(T) should be classified as representing a novel species in the genus Kiloniella and the name proposed is Kiloniella spongiae sp. nov. The type strain is MEBiC09566(T) ( = KCCM 43040(T) = JCM 19930(T)). Emended descriptions of the genus Kiloniella Wiese et al. 2009 and Kiloniella laminariae are also given.

Topics: Alphaproteobacteria; Animals; Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Molecular Sequence Data; Phospholipids; Phylogeny; Porifera; Republic of Korea; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone

A Gram-staining-negative, rod-shaped, motile and facultatively anaerobic bacterial strain, designated X2(T), was isolated from the sludge of an anaerobic, denitrifying, sulfide-removal bioreactor, and found to oxidize sulfide anaerobically with nitrate as electron acceptor. The strain grew at salinities of 0-3% (w/v) NaCl (optimum, 0-1%). Growth occurred at pH 6.0-10.0 (optimum, pH 8.0) and 10-37 °C (optimum, 30 °C). The genomic DNA G+C content was 59 mol%. Q-8 and Q-9 were detected as the respiratory quinones. The major fatty acids (>10 %) were C16:1ω7c and/or C16: 1ω6c, C18: 1ω7c and C16:0. The polar lipids consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and one unidentified phospholipid. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain X2(T) formed a novel clade within the family Pseudomonadaceae, with the highest sequence similarity to Pseudomonas caeni KCTC 22292(T) (93.5%). On the basis of phenotypic, chemotaxonomic and phylogenetic characteristics, it is proposed that this strain represents novel genus and species within the family Pseudomonadaceae, for which the name Thiopseudomonas denitrificans gen. nov., sp. nov. is proposed. The type strain is X2(T) ( =CCTCC M 2013362(T) =DSM 28679(T) = KCTC 42076(T)).

Topics: Bacterial Typing Techniques; Base Composition; Bioreactors; DNA, Bacterial; Fatty Acids; Molecular Sequence Data; Phospholipids; Phylogeny; Pseudomonadaceae; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Sewage; Ubiquinone

A psychrotolerant strain, 8H1(T), was isolated from soil samples collected in Isla de los Estados, Ushuaia, Argentina. Cells were Gram-negative, aerobic, straight rods, occurring singly or in pairs, non-spore-forming and motile by means of two polar flagella. The isolate was able to grow in the range 4-35 °C, with optimum growth at 28 °C. The predominant cellular fatty acids were summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c), C16 : 0 and summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c). The polar lipid pattern of strain 8H1(T) comprised phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and an unknown phospholipid. Ubiquinone 9 (Q-9) was the predominant lipoquinone. The DNA G+C content was 59.8 mol%. 16S rRNA gene sequence-based phylogeny suggested the affiliation of strain 8H1(T) to the 'Pseudomonas fluorescens group', displaying ≥98.5 % sequence similarity to 29 type strains. A multilocus sequence analysis (MLSA) study performed by concatenating 16S rRNA, gyrB, rpoD and rpoB gene sequences showed that isolate 8H1(T) could be discriminated from closely related species of the genus Pseudomonas and placed in the 'Pseudomonas gessardii subgroup', including the species with the highest MLSA sequence similarities: Pseudomonas brenneri (96.2 %), P. gessardii (96.1 %), P. proteolytica (96.0 %), P. meridiana (96.0 %) and P. mucidolens (95.4 %). DNA-DNA hybridization analysis between 8H1(T) and the type strains of these closely related species revealed relatedness values of 27.0, 8.8, 41.2, 39.7 and 46.1 %, respectively. These results, together with differences in several phenotypic features, support the classification of a novel species, for which the name Pseudomonas yamanorum sp. nov. is proposed. The type strain is 8H1(T) ( = DSM 26522(T) = CCUG 63249(T) = LMG 27247(T)).

Topics: Argentina; Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Genes, Bacterial; Molecular Sequence Data; Multilocus Sequence Typing; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Pseudomonas; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Soil Microbiology; Ubiquinone

An aerobic, Gram-stain-negative, rod-shaped and polar-flagellated bacterium, designated strain CC-MHH0089(T), was isolated from a soil sample taken on Matsu Island (Taiwan). Strain CC-MHH0089(T) grew at 15-30 °C and pH 5.0-10.0 and tolerated ≤8 % (w/v) NaCl. 16S rRNA gene sequence analysis showed high pairwise sequence similarity to Pseudomonas azotifigens 6H33b(T) (97.3 %) and Pseudomonas balearica SP1402(T) (96.7 %) and lower sequence similarity to other strains (<96.0 %). In DNA-DNA reassociation experiments, the relatedness of strain CC-MHH0089(T) to P. azotifigens JCM 12708(T) was 38.3 % (reciprocal value 19.5 %). Evolutionary trees reconstructed on the basis of 16S rRNA, gyrB and rpoB gene sequences revealed a varying phylogenetic neighbourhood of strain CC-MHH0089(T) with regard to the most closely related type strains. The predominant quinone system was ubiquinone 9 (Q-9) and the DNA G+C content was 63.6 mol%. The major fatty acids were C12 : 0, C16 : 0, C17 : 0, C19 : 0 cyclo ω8c and summed features 2 (C14 : 0 3-OH/iso-C16 : 1 I), 3 (C16 : 1ω7c/C16 : 1ω6c) and 8 (C18 : 1ω7c/C18 : 1ω6c). The major polar lipids were phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine and diphosphatidylglycerol. According to its distinct phylogenetic, phenotypic and chemotaxonomic features, strain CC-MHH0089(T) is proposed to represent a novel species within the genus Pseudomonas, for which the name Pseudomonas matsuisoli sp. nov. is proposed. The type strain is CC-MHH0089(T) ( = BCRC 80771(T) = JCM 30078(T)).

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Genes, Bacterial; Molecular Sequence Data; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Pseudomonas; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Soil Microbiology; Taiwan; Ubiquinone

Strain NEAU-ST5-21(T) was isolated from saline and alkaline soils in Zhaodong City, Heilongjiang Province, China. It was aerobic, Gram-stain-negative, rod-shaped and motile with a polar flagellum. It produced yellow-orange colonies with a smooth surface, and grew in the presence of 0-5 % (w/v) NaCl (optimum 0 %, w/v), at temperatures of 20-40 °C (optimum 28 °C) and at pH 7-11 (optimum pH 7). Phylogenetic analyses based on the separate 16S rRNA gene sequences and concatenated 16S rRNA, gyrB and rpoD gene sequences indicated that strain NEAU-ST5-21(T) belongs to the genus Pseudomonas in the class Gammaproteobacteria. The most closely related species is Pseudomonas xanthomarina, whose type strain (KMM 1447(T)) showed gene sequence similarities of 99.0 % for 16S rRNA, 81.8 % for gyrB and 85.0 % for rpoD with strain NEAU-ST5-21(T). DNA-DNA hybridization values between strain NEAU-ST5-21(T) and P. xanthomarina DSM 18231(T), Pseudomonas kunmingensis CGMCC 1.12273(T), Pseudomonas stutzeri DSM 5190(T), Pseudomonas oleovorans subsp. lubricantis DSM 21016(T), Pseudomomas chengduensis CGMCC 2318(T), Pseudomonas alcaliphila DSM 17744(T) and Pseudomonas toyotomiensis DSM 26169(T) were 52±0 % to 25±2 %. The DNA G+C content of strain NEAU-ST5-21(T) was 65 mol%. The major fatty acids (>10 %) were C18 : 1ω7c and/or C18 : 1ω6c, C16 : 1ω7c and/or C16 : 1ω6c and C16 : 0, the predominant respiratory quinone was ubiquinone 9, and polar lipids consisted of diphosphatidylglycerol, phosphatidylethanolamine, one unknown phospholipid, phosphatidylglycerol, one unknown aminolipid, one unknown lipid and a glycolipid. The proposed name is Pseudomonas zhaodongensis sp. nov., NEAU-ST5-21(T) ( = ACCC 06362(T) = DSM 27559(T)) being the type strain.

Topics: Bacterial Typing Techniques; Base Composition; China; DNA, Bacterial; Fatty Acids; Genes, Bacterial; Hydrogen-Ion Concentration; Molecular Sequence Data; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Pigmentation; Pseudomonas; RNA, Ribosomal, 16S; Salinity; Sequence Analysis, DNA; Soil; Soil Microbiology; Ubiquinone

The aim of study was to search for new biomarkers with a magnetic resonance technique to identify the early stages of dementia, induced by D-galactose, and evaluate Simvastatin therapy. Localized proton magnetic resonance spectroscopy measurements showed a significant decrease in the concentration of N-acetylaspartate+N-acetylaspartylglutamate and myo-inositol in the D-galactose group compared to the control group, and, conversely, an increase of N-acetylaspartate+N-acetylaspartylglutamate in the D-galactose/Simvastatin group. Using a saturation transfer experiment, with phosphorus magnetic resonance spectroscopy, we observed a significant elevation of the forward rate constant of the creatine kinase reaction in the brains of the D-galactose group compared to controls, and subsequently, a significant reduction of this reaction in the D-galactose/Simvastatin group. Spatial learning and memory were evaluated using the modified Morris water maze test. The dynamics of the learning process represented by the learning index revealed a significant reduction in learning in the D-galactose group, but the deficits as a consequence of the D-galactose effects were recovered in the D-galactose/Simvastatin group, in which the learning dynamics resembled those of the control group. By determining the thiobarbituric acid reactive substances and total coenzyme Q9 in plasma, we have shown that long-term administration of D-galactose created conditions for oxidative stress, and that the administration of Simvastatin decreased oxidative stress in plasma. Volumetry analyses from the hippocampal area show a reduction in the segmented area in the D-galactose group, compared with the control group, and an enlarged area in the hippocampus in the d-galactose/Simvastatin group.

Topics: Animals; Aspartic Acid; Biomarkers; Brain; Dementia; Dipeptides; Disease Models, Animal; Galactose; Inositol; Magnetic Resonance Spectroscopy; Male; Nootropic Agents; Organ Size; Phosphorus Isotopes; Protons; Rats, Wistar; Simvastatin; Spatial Learning; Spatial Memory; Thiobarbituric Acid Reactive Substances; Treatment Outcome; Ubiquinone

An aerobic, Gram-negative, motile, rod-shaped bacterium designated KMM 9500(T) was isolated from a sediment sample collected from the Sea of Japan seashore. Comparative 16S rRNA gene sequence analysis affiliated strain KMM 9500(T) to the genus Pseudomonas as a distinct subline clustered with Pseudomonas marincola KMM 3042(T) and Pseudomonas segetis KCTC 12331(T) sharing the highest similarities of 98 and 97.9 %, respectively. Strain KMM 9500(T) was characterized by mainly possessing ubiquinone Q-9, and by the predominance of C18:1 ω7c, C16:1 ω7c, and C16:0 followed by C12:0 in its fatty acid profile. Polar lipids consisted of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, an unknown aminophospholipid, and unknown phospholipids. Strain KMM 9500(T) was found to inhibit growth of Gram-negative and Gram-positive indicatory microorganisms. Based on the phylogenetic analysis and distinctive phenotypic characteristics, strain 9500(T) is concluded to represent a novel species of the genus Pseudomonas, for which the name Pseudomonas glareae sp. nov. is proposed. The type strain of the species is strain KMM 9500(T) (=NRIC 0939(T)).

Topics: Antibiosis; Bacterial Typing Techniques; DNA, Bacterial; Fatty Acids; Geologic Sediments; Japan; Lipids; Oceans and Seas; Phospholipids; Phylogeny; Pseudomonas; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone

Primary coenzyme Q10 (CoQ10) deficiency is due to mutations in genes involved in CoQ biosynthesis. The disease has been associated with five major phenotypes, but a genotype-phenotype correlation is unclear. Here, we compare two mouse models with a genetic modification in Coq9 gene (Coq9(Q95X) and Coq9(R239X)), and their responses to 2,4-dihydroxybenzoic acid (2,4-diHB). Coq9(R239X) mice manifest severe widespread CoQ deficiency associated with fatal encephalomyopathy and respond to 2,4-diHB increasing CoQ levels. In contrast, Coq9(Q95X) mice exhibit mild CoQ deficiency manifesting with reduction in CI+III activity and mitochondrial respiration in skeletal muscle, and late-onset mild mitochondrial myopathy, which does not respond to 2,4-diHB. We show that these differences are due to the levels of COQ biosynthetic proteins, suggesting that the presence of a truncated version of COQ9 protein in Coq9(R239X) mice destabilizes the CoQ multiprotein complex. Our study points out the importance of the multiprotein complex for CoQ biosynthesis in mammals, which may provide new insights to understand the genotype-phenotype heterogeneity associated with human CoQ deficiency and may have a potential impact on the treatment of this mitochondrial disorder.

Topics: Animals; Ataxia; Disease Models, Animal; Genetic Variation; Genotype; Hydroxybenzoates; Mammals; Mice; Mice, Transgenic; Mitochondrial Diseases; Muscle Weakness; Mutation, Missense; Ubiquinone

Today HIV-1 infection is recognized as a chronic disease with obligatory lifelong treatment to keep viral titers below detectable levels. The continuous intake of antiretroviral drugs however, leads to severe and even life-threatening side effects, supposedly by the deleterious impact of nucleoside-analogue type compounds on the functioning of the mitochondrial DNA polymerase. For detailed investigation of the yet partially understood underlying mechanisms, the availability of a versatile model system is crucial. We therefore set out to develop the use of Caenorhabditis elegans to study drug induced mitochondrial toxicity. Using a combination of molecular-biological and functional assays, combined with a quantitative analysis of mitochondrial network morphology, we conclude that anti-retroviral drugs with similar working mechanisms can be classified into distinct groups based on their effects on mitochondrial morphology and biochemistry. Additionally we show that mitochondrial toxicity of antiretroviral drugs cannot be exclusively attributed to interference with the mitochondrial DNA polymerase.

Topics: Animals; Anti-HIV Agents; Caenorhabditis elegans; Didanosine; Dideoxynucleosides; DNA-Directed DNA Polymerase; DNA, Mitochondrial; Drug Evaluation; Humans; Mitochondria; Mitochondrial Proteins; Models, Biological; Oxygen Consumption; Reverse Transcriptase Inhibitors; Stavudine; Ubiquinone; Zalcitabine; Zidovudine

A Gram-staining-negative, facultatively aerobic bacterium, strain XCD-X85(T), was isolated from Xiaochaidan Lake, a salt lake (salinity 9.9%, w/v) in Qaidam basin, Qinghai province, China. Its taxonomic position was determined by using a polyphasic approach. Cells of strain XCD-X85(T) were non-endospore-forming rods, 0.4-0.6 μm wide and 1.0-1.6 μm long, and motile by means of a single polar flagellum. Strain XCD-X85(T) was catalase- and oxidase-positive. Growth was observed in the presence of 0-12.0% (w/v) NaCl (optimum, 1.0-2.0%) and at 4-35 °C (optimum, 25-30 °C) and pH 6.5-10.5 (optimum, pH 8.0-8.5). Strain XCD-X85(T) contained (>10%) summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c), C12 : 0, C16 : 0 and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) as the predominant fatty acids. The major respiratory quinone was ubiquinone 9 (Q-9). The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The DNA G+C content was 57.4 mol%. Phylogenetic trees based on 16S rRNA gene sequences showed that strain XCD-X85(T) was associated with the genus Pseudomonas, and showed highest 16S rRNA gene sequence similarities to Pseudomonas pelagia CL-AP6(T) (99.0%) and Pseudomonas bauzanensis BZ93(T) (96.8%). DNA-DNA relatedness of strain XCD-X85T to P. pelagia JCM 15562(T) was 19 ± 1%. On the basis of the data presented above, it is concluded that strain XCD-X85(T) represents a novel species of the genus Pseudomonas, for which the name Pseudomonas salina sp. nov. is proposed. The type strain is XCD-X85(T) ( = CGMCC 1.12482(T) = JCM 19469(T)).

Topics: Bacterial Typing Techniques; Base Composition; China; DNA, Bacterial; Fatty Acids; Lakes; Molecular Sequence Data; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Pseudomonas; RNA, Ribosomal, 16S; Salinity; Sequence Analysis, DNA; Ubiquinone; Water Microbiology

We examined how dietary supplementation of vitamin E protects against liver oxidative damage in rats with water-immersion restraint stress (WIRS). Before WIRS exposure, rats received a normal diet (ND) or vitamin E-supplemented diet (VESD) (500 IU α-tocopherol/kg diet) at a mean dose of 15 g/animal/d for 4 wk. The two diet groups had serum transaminases and lactate dehydrogenase activities and adrenocorticotropic hormone, corticosterone, and glucose levels to a similar extent. VESD-fed rats had higher liver α-tocopherol concentrations and lower liver ascorbic acid, total coenzyme Q9 (CoQ9), reduced CoQ9, reduced CoQ10, and lipid peroxide (LPO) concentrations than ND-fed rats. When the two diet groups were exposed to 6 h of WIRS, the serum liver cell damage index enzyme activities increased more greatly in ND-fed rats than in VESD-fed rats but the serum stress marker levels increased to a similar extent. The WIRS exposure caused no change in liver LPO concentration with the further increase in liver α-tocopherol concentration in VESD-fed rats but increased liver LPO concentration without changing liver α-tocopherol concentration in ND-fed rats. Upon the WIRS exposure, liver reduced glutathione concentration decreased with the further decrease in liver ascorbic acid concentration in VESD-fed rats and those concentrations decreased in ND-fed rats. The WIRS exposure recovered the decreased liver total CoQ9 and reduced CoQ9 concentrations in VESD-fed rats but decreased liver total CoQ9, reduced CoQ9, and reduced CoQ10 concentrations in ND-fed rats. These results indicate that dietary vitamin E supplementation protects against liver oxidative damage without affecting the stress response in rats with WIRS.

Topics: alpha-Tocopherol; Animals; Antioxidants; Ascorbic Acid; Biomarkers; Diet; Dietary Supplements; Glutathione; Immersion; Lipid Peroxidation; Lipid Peroxides; Liver; Male; Oxidative Stress; Rats, Sprague-Dawley; Restraint, Physical; Stress, Physiological; Thiobarbituric Acid Reactive Substances; Ubiquinone; Vitamin E

A Gram-negative, helical bacterium designated PH27AT was cultivated from an anchialine pool on Pearl and Hermes Atoll, Northwestern Hawaiian Islands. The obligately halophilic strain was motile by bipolar tufts of flagella and grew optimally at pH 7, and microaerobically or aerobically. Closest neighbours based on 16S rRNA gene nucleotide sequence identity are Marinospirillum celere v1c_Sn-redT (93.31 %) and M. alkaliphilum Z4T (92.10 %) in the family Oceanospirillaceae, class Gammaproteobacteria. PH27AT is distinguished phenotypically from members of the genus Marinospirillum by its hydrolysis of gelatin, the absence of growth in media containing ≤ 1 % (w/v) NaCl and the ranges of temperature (12–40 °C) and pH (5–8) for growth. The major compound ubiquinone Q-9 distinguishes the quinone system of strain PH27AT from those in members of the genus Marinospirillum and other members of the Oceanospirillaceae, in which the major quinone is Q-8. Major polar lipids in PH27AT were phosphatidylethanolamine and phosphatidylglycerol, with moderate amounts of diphosphatidylglycerol and phosphatidylserine. Spermidine and cadaverine dominated the polyamine pattern; large proportions of cadaverine have not been reported in members of the genus Marinospirillum. Genotypic and chemotaxonomic data show that PH27AT does not belong in the genus Marinospirillum or other genera of the family Oceanospirillaceae or the Halomonadaceae. We propose a new genus, Terasakiispira gen. nov., be created to accommodate Terasakiispira papahanaumokuakeensis gen. nov., sp. nov. as the type species, with PH27AT ( = ATCC BAA-995T = DSM 16455T = DSM 23961T) as the type strain.

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Flagella; Gammaproteobacteria; Hawaii; Islands; Molecular Sequence Data; Phospholipids; Phylogeny; Polyamines; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone; Water Microbiology

A red-pigmented bacterium producing a metallic green sheen, designated strain JC333T, was isolated from a sand sample collected from Shivrajpur-Kachigad beach, Gujarat, India. Phylogenetic analyses based on the 16S rRNA gene sequence of strain JC333T showed highest sequence similarity to Zooshikella ganghwensis JC2044T (99.24 %) and less than 91.94 % similarity with other members of the class Gammaproteobacteria. DNA-DNA hybridizations between JC333T and Z. ganghwensis JC2044T showed low relatedness values of 19 ± 1.3 % (reciprocal 21 ± 2.2 %). The major respiratory quinone was ubiquinone-9 (Q9) and the polar lipid profile was composed of the major components diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, an unidentified aminophospholipid and an unidentified lipid. The presence of C16 : 1ω7c/C16 : 1ω6c, C16 : 0, C18 : 1ω7c and C12 : 0 as major fatty acids supported the affiliation of strain JC333T to the genus Zooshikella. Prodigiosin, cycloprodigiosin and eight other prodigiosin analogues were the pigments of JC333T. Characterization based on 16S rRNA gene sequence analysis, physiological parameters, pigment analysis, ubiquinone, and polar lipid and fatty acid compositions revealed that JC333T represents a novel species of the genus Zooshikella, for which the name Zooshikella marina sp. nov. is proposed. The type strain is JC333T ( = KCTC 42659T = LMG 28823T).

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Gammaproteobacteria; India; Indoles; Molecular Sequence Data; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Pigmentation; Prodigiosin; Pyrroles; RNA, Ribosomal, 16S; Seawater; Sequence Analysis, DNA; Ubiquinone

A Gram-stain-negative, halotolerant and alkalitolerant bacterium, designated strain BH103T, was isolated from saltern soil in Gomso, Korea. Cells of strain BH103T were strictly aerobic, motile, straight rods and grew at pH 7.0-10.8 (optimum, pH 8.5), at 10-55 °C (optimum, 28 °C) and at salinities of 0-23 % (w/v) NaCl (optimum, 14 % NaCl). Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain BH103T belongs to the genus Halomonas, showing highest sequence similarity to Halomonas boliviensis LC1T (97.7 %), Halomonas neptunia Eplume1T (97.7 %), Halomonas variabilis IIIT (97.7 %), Halomonas alkaliantarctica CRSST (97.7 %), Halomonas olivaria TYRC17T (97.5 %), Halomonas titanicae BH1T (97.2 %) and Halomonas sulfidaeris Esulfide1T (96.2 %). The predominant ubiquinone was Q-9. The major fatty acids were C18 : 1ω7c, C16 : 1ω7c and/or iso-C15 : 0 2-OH, C16 : 0 and C12 : 0 3-OH. The major polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, aminophospholipid and an unknown phospholipid. The DNA G+C content of this novel isolate was 54.7 mol%. DNA-DNA relatedness between strain BH103T and H. boliviensis KACC 16615T, H. neptunia KCTC 2888T, H. variabilis KCTC 2889T, H. alkaliantarctica KCTC 22844T, H. olivaria DSM 19074T, H. titanicae JCM 16411T and H. sulfidaeris DSM 15722T was 45, 41, 39, 32, 38, 45 and 35 %, respectively. On the basis of polyphasic analysis from this study, strain BH103T represents a novel species of the genus Halomonas, for which the name Halomonas salicampi sp. nov. is proposed. The type strain is BH103T ( = KACC 17609T = NBRC 109914T = NCAIM B 02528T).

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Halomonas; Molecular Sequence Data; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Republic of Korea; RNA, Ribosomal, 16S; Salinity; Sequence Analysis, DNA; Soil Microbiology; Ubiquinone

A Gram-stain-negative, motile, aerobic and rod-shaped bacterium, designated HJM-18T, was isolated from the place where the ocean and a freshwater lake meet at Hwajinpo, South Korea, and subjected to a taxonomic study using a polyphasic approach. Strain HJM-18T grew optimally at 30 °C, at pH 7.0-8.0 and in the presence of 1.0-3.0 % (w/v) NaCl. Phylogenetic trees based on 16S rRNA gene sequences showed that strain HJM-18T belonged to the genus Marinobacter. Strain HJM-18T exhibited 16S rRNA gene sequence similarity values of 97.05-98.22 % to the type strains of Marinobacter algicola, Marinobacter flavimaris, Marinobacter adhaerens, Marinobacter salarius, Marinobacter salsuginis, Marinobacter guineae and Marinobacter gudaonensis and of 93.21-96.98 % to the type strains of the other species of the genus Marinobacter. Strain HJM-18T contained Q-9 as the predominant ubiquinone and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), C16 : 0 and C18 : 1ω9c as the major fatty acids. The major polar lipids detected in strain HJM-18T were phosphatidylethanolamine, phosphatidylglycerol and one unidentified aminophospholipid. The DNA G+C content was 58 mol% and the mean DNA-DNA relatedness values with the type strains of the seven phylogenetically related species of the genus Marinobacter were 10-27 %. Differential phenotypic properties, together with phylogenetic and genetic distinctiveness, revealed that strain HJM-18T is separated from recognized species of the genus Marinobacter. On the basis of the data presented, strain HJM-18T represents a novel species of the genus Marinobacter, for which the name Marinobacter confluentis sp. nov. is proposed. The type strain is HJM-18T ( = KCTC 42705T = NBRC 111223T).

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Fresh Water; Lakes; Marinobacter; Molecular Sequence Data; Nucleic Acid Hybridization; Oceans and Seas; Phospholipids; Phylogeny; Republic of Korea; RNA, Ribosomal, 16S; Seawater; Sequence Analysis, DNA; Ubiquinone

Strain MBR(T) was isolated from landfill leachate in a solid-waste disposal site in Chengdu, Sichuan, China. An analysis of 16S rRNA gene sequences revealed that the isolate was closely related to members of the genus Pseudomonas, sharing the highest sequence similarities with Pseudomonas toyotomiensis HT-3(T) (99.8 %), Pseudomonas alcaliphila AL15-21(T) (99.7 %) and Pseudomonas oleovorans ATCC 8062(T) (99.4 %). Multi-locus sequence analysis based on three housekeeping genes (gyrB, rpoB and rpoD) provided higher resolution at the species level than that based on 16S rRNA gene sequences, which was further confirmed by less than 70 % DNA-DNA relatedness between the new isolate and P. toyotomiensis HT-3(T) (61.3 %), P. alcaliphila AL15-21(T) (51.5 %) and P. oleovorans ATCC 8062(T) (57.8 %). The DNA G+C content of strain MBR(T) was 61.9 mol% and the major ubiquinone was Q-9. The major cellular fatty acids (>10 %) were C18 : 1ω7c and/or C18 : 1ω6c, C16 : 0, and C16 : 1ω7c and/or C16 : 1ω6c. Polyphasic analysis indicates that strain MBR(T) represents a novel species of the genus Pseudomonas, for which the name Pseudomonas chengduensis sp. nov. is proposed. The type strain is MBR(T) ( = CGMCC 2318(T) = DSM 26382(T)).

Topics: Bacterial Typing Techniques; Base Composition; China; DNA, Bacterial; Fatty Acids; Genes, Bacterial; Molecular Sequence Data; Nucleic Acid Hybridization; Phylogeny; Pseudomonas; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Solid Waste; Ubiquinone; Waste Disposal Facilities; Water Microbiology; Water Pollutants, Chemical

A moderately halophilic, Gram-stain-negative, non-sporulating bacterium designed as strain TYRC17(T) was isolated from olive-processing effluents. The organism was a straight rod, motile by means of peritrichous flagella and able to respire both oxygen and nitrate. Growth occurred with 0-25 % (w/v) NaCl (optimum, 7 %), at pH 5-11 (optimum, pH 7.0) and at 4-50 °C (optimally at 35 °C). It accumulated poly-β-hydroxyalkanoate granules and produced exopolysaccharides. The predominant fatty acids were C18 : 1ω7c, C16 : 1ω7c and C16 : 0. Ubiquinone 9 (Q-9) was the only respiratory quinone. The DNA G+C content of TYRC17(T) was 53.9 mol%. Phylogenetic analyses of 16S rRNA gene sequences revealed that the strain represents a member of the genus Halomonas and more precisely of the subgroup containing Halomonas sulfidaeris, H. titanicae, H. variabilis, H. zhanjiangensis, H. alkaliantarctica, H. boliviensis and H. neptunia. TYRC17(T) showed high 16S-rRNA sequence identities in particular with the three last species listed (99.4-99.5 %). A multilocus sequence analysis (MLSA) using the 23S rRNA, gyrB, rpoD and secA genes allowed clarifying the phylogenetic position of TYRC17(T). This, combined with the level of DNA-DNA hybridization between TYRC17(T) and its closest relatives ranging from 21.6 % to 48.4 %, indicated that TYRC17(T) did not represent any of these species. On the basis of phenotypic and genotypic characteristics, and also genomic and phylogenetic evidence, it was concluded that strain TYRC17(T) represented a novel species of the genus Halomonas. The name Halomonas olivaria sp. nov. is proposed with TYRC17(T) ( = DSM 19074(T) = CCUG 53850B(T)) as the type strain.

Topics: Bacterial Typing Techniques; Base Composition; Bayes Theorem; DNA, Bacterial; Fatty Acids; Genes, Bacterial; Halomonas; Hydroxybutyrates; Molecular Sequence Data; Morocco; Multilocus Sequence Typing; Nucleic Acid Hybridization; Olea; Phylogeny; Polyesters; Polysaccharides, Bacterial; RNA, Ribosomal, 16S; Ubiquinone; Wastewater

A Gram-stain-negative, rod-shaped, exopolysaccharide-producing, strictly aerobic bacterium with a single polar flagellum, designated strain HL22-2(T), was isolated from a phosphate mine situated in a suburb of Kunmming in Yunnan province in south-western China. The taxonomic status of this strain was evaluated by using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain HL22-2(T) was related to members of the genus Pseudomonas. 16S rRNA gene sequence similarities between strain HL22-2(T) and Pseudomonas xanthomarina KMM 1447(T), Pseudomonas alcaliphila AL15-21(T) and Pseudomonas stutzeri ATCC 17588(T) were 98.9, 98.10% and 98.06%, respectively. The major cellular fatty acids were C(18 : 1)ω7c, C(16 : 0) and summed feature 3 (C(16 : 1)ω7c and/or C(16 : 1)ω6c). The DNA G+C content was 60.3 mol%. On the basis of phenotypic characteristics, phylogenetic analysis and DNA-DNA relatedness values, strain HL22-2(T) represents a novel species of the genus Pseudomonas, for which the name Pseudomonas kunmingensis sp. nov. is proposed. The type strain is HL22-2(T) ( = CGMCC 1.12273(T) = DSM 25974(T)).

Topics: Bacterial Typing Techniques; Base Composition; China; DNA, Bacterial; Fatty Acids; Mining; Molecular Sequence Data; Nucleic Acid Hybridization; Phosphates; Phylogeny; Pseudomonas; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Soil Microbiology; Ubiquinone

A novel, Gram-stain-negative, aerobic, rod-shaped, non-motile and moderately halophilic bacterium, designated strain BJGMM-B45(T), was isolated from a saline-alkali soil collected from Shandong Province, China. Growth of strain BJGMM-B45(T) occurred at 10-45 °C (optimum, 30 °C) and pH 5.0-12.0 (optimum, pH 7.0) on Luria-Bertani agar medium with 1-20 % (w/v) NaCl (optimum, 7-10 %). The predominant respiratory quinone was Q-9. The major cellular fatty acids (>5 %) were C18 : 1ω7c, C16 : 0, C19 : 0 cyclo ω8c, summed feature 3, C12 : 0 3-OH and C12 : 0. The genomic DNA G+C content was 57.5 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain BJGMM-B45(T) belonged to the genus Halomonas in the class Gammaproteobacteria. The closest relatives were Halomonas cupida DSM 4740(T) (98.2 % 16S rRNA gene sequence similarity) and Halomonas denitrificans M29(T) (97.8 %). Levels of DNA-DNA relatedness between strain BJGMM-B45(T) and Halomonas cupida CGMCC 1.2312(T) and Halomonas denitrificans DSM 18045(T) were 57.0 and 58.9 %, respectively. On the basis of phenotypic, chemotaxonomic and phylogenetic features, strain BJGMM-B45(T) is considered to represent a novel species of the genus Halomonas, for which the name Halomonas huangheensis sp. nov. is proposed. The type strain is BJGMM-B45(T) ( = ACCC 05850(T) = KCTC 32409(T)).

Topics: Alkalies; Base Composition; China; DNA, Bacterial; Fatty Acids; Halomonas; Molecular Sequence Data; Nucleic Acid Hybridization; Phylogeny; RNA, Ribosomal, 16S; Salinity; Soil Microbiology; Ubiquinone

A moderately halophilic bacterium (strain NEAU-ST10-39T) was isolated from saline and alkaline soils in the oilfield of Daqing City, Heilongjiang Province, China. The strain was strictly aerobic, Gram-stain-negative, rod-shaped and motile by peritrichous flagella. Its colonies were yellow. It grew at NaCl concentrations of 0.2-15% (w/v) (optimum 4%, w/v), at temperatures of 4-40 °C (optimum 35 °C) and at pH 5-10 (optimum pH 7). It did not produce acids from sugars or alcohols. Its DNA G+C content was 57.4 mol%. Phylogenetic analyses based on 16S rRNA gene sequences and concatenated 16S rRNA, gyrB and rpoD gene sequences indicated that it belonged to the genus Halomonas in the class Gammaproteobacteria. The most phylogenetically related species were Halomonas axialensis, Halomonas meridiana and Halomonas aquamarina, whose types shared 98.3% (16S rRNA), 82.7% (gyrB) and 83.9-84.5% (rpoD) sequence similarity with strain NEAU-ST10-39T. The results of DNA-DNA hybridization assays showed 20±2%-50±1 % relatedness between strain NEAU-ST10-39T and the most closely related species including Halomonas axialensis DSM 15723T, Halomonas meridiana DSM 5425T, Halomonas aquamarina DSM 30161(T), Halomonas johnsoniae DSM 21197T, Halomonas stevensii DSM 21198T, Halomonas nanhaiensis CCTCC AB 2012911(T), Halomonas hamiltonii DSM 21196T and Halomonas arcis CGMCC 1.6494T. The major fatty acids were C18 : 1ω7c (47.2%), C16:1ω7c and/or C16:1ω6c (18.9%) and C16:0 (16.3%), the only respiratory quinone detected was ubiquinone 9 and polar lipids consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, two unknown phospholipids and three unknown lipids. The new isolate is proposed to represent a novel species with the name Halomonas songnenensis sp. nov., NEAU-ST10-39T (=CGMCC 1.12152T=DSM 25870T) being the type strain.

Topics: Bacterial Typing Techniques; Base Composition; China; DNA, Bacterial; Fatty Acids; Genes, Bacterial; Halomonas; Hydrogen-Ion Concentration; Molecular Sequence Data; Nucleic Acid Hybridization; Phospholipids; Phylogeny; RNA, Ribosomal, 16S; Salinity; Sequence Analysis, DNA; Soil; Soil Microbiology; Ubiquinone

A novel bacterium, designated strain WP70(T), was isolated from the gut of a comb pen shell (Atrina pectinata) collected from the southern sea of Yeosu in Korea. The isolate was Gram-stain-negative, aerobic, non-motile and rod-shaped. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain WP70(T) belonged to the genus Endozoicomonas. The highest level of sequence similarity (98.4%) was shared with Endozoicomonas elysicola MKT110(T). Optimal growth occurred in 2% (w/v) NaCl at 30 °C and at pH 7. The major cellular fatty acids were summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c) and C16 : 0. The main respiratory quinone was Q-9. The polar lipids comprised phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol, three unidentified phospholipids, an unidentified aminolipid, an unidentified aminophospholipid and an unidentified lipid. The genomic DNA G+C content was 50.5 mol% and DNA-DNA hybridization values indicated <11% genomic relatedness to the closest species. Physiological, biochemical, chemotaxonomic and genotypic analyses indicated that strain WP70(T) represents a novel species of the genus Endozoicomonas, for which the name Endozoicomonas atrinae sp. nov. is proposed. The type strain is WP70(T) ( = KACC 17474(T)  = JCM 19190(T)).

Topics: Animals; Bacterial Typing Techniques; Base Composition; Bivalvia; DNA, Bacterial; Fatty Acids; Gammaproteobacteria; Gastrointestinal Tract; Molecular Sequence Data; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Republic of Korea; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone

A bacterial strain, OHA11(T), was isolated during the course of a study of phosphate-solubilizing bacteria occurring in a forest soil from Salamanca, Spain. The 16S rRNA gene sequence of strain OHA11(T) shared 99.1% similarity with respect to Pseudomonas baetica a390(T), and 98.9% similarity with the type strains of Pseudomonas jessenii, Pseudomonas moorei, Pseudomonas umsongensis, Pseudomonas mohnii and Pseudomonas koreensis. The analysis of housekeeping genes rpoB, rpoD and gyrB confirmed its phylogenetic affiliation to the genus Pseudomonas and showed similarities lower than 95% in almost all cases with respect to the above species. Cells possessed two polar flagella. The respiratory quinone was Q9. The major fatty acids were C16 : 0, C18 : 1ω7c and summed feature 3 (C16 : 1ω7c/iso-C15 : 0 2-OH). The strain was oxidase-, catalase- and urease-positive, positive for arginine dihydrolase but negative for nitrate reduction, β-galactosidase production and aesculin hydrolysis. It was able to grow at 31 °C and at pH 11. The DNA G+C content was 58.1 mol%. DNA-DNA hybridization results showed values lower than 49% relatedness with respect to the type strains of the seven closest related species. Therefore, the combined genotypic, phenotypic and chemotaxonomic data support the classification of strain OHA11(T) to a novel species of the genus Pseudomonas, for which the name Pseudomonas helmanticensis sp. nov. is proposed. The type strain is OHA11(T) ( = LMG 28168(T) = CECT 8548(T)).

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Genes, Bacterial; Molecular Sequence Data; Nucleic Acid Hybridization; Phylogeny; Pseudomonas; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Soil Microbiology; Spain; Trees; Ubiquinone

Two Gram-staining-negative, aerobic, rod-shaped, non-spore-forming bacterial strains that are motile by a monopolar flagellum, designated CC-AMH-11(T) and CC-AMHZ-5, were isolated from droppings of a seashore bird off the coast of Hualien, Taiwan. The strains showed 99.7% mutual pairwise 16S rRNA gene sequence similarity, while exhibiting <96.2% sequence similarity to strains of other species of the genus Pseudomonas (95.7-95.9% similarity with type species, Pseudomonas aeruginosa LMG 1242T), and formed a distinct co-phyletic lineage in the phylogenetic trees. The common major fatty acids (>5% of the total) were C18 : 1ω7c and/or C18 : 1ω6c (summed feature 8), C16 : 1ω6c and/or C16 : 1ω7c (summed feature 3), C16 : 0 and C12 : 0. Phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylserine, an unidentified lipid and an unidentified phospholipid were detected as common polar lipids. The DNA G+C contents of strains CC-AMH-11(T) and CC-AMHZ-5 were 61.1 and 61.6 mol%, respectively. The common major respiratory quinone was ubiquinone 9 (Q-9), and the predominant polyamine was putrescine. The DNA-DNA hybridization obtained between the two strains was 79.0% (reciprocal value 89.4% using CC-AMHZ-5 DNA as the probe). The very high 16S rRNA gene sequence similarity and DNA-DNA relatedness and the poorly distinguishable phenotypic features witnessed between CC-AMH-11(T) and CC-AMHZ-5 suggested unambiguously that they are two distinct strains of a single genomic species. However, the strains also showed several genotypic and phenotypic characteristics that distinguished them from other closely related species of Pseudomonas. Thus, the strains are proposed to represent a novel species of Pseudomonas, for which the name Pseudomonas hussainii sp. nov. is proposed. The type strain is CC-AMH-11(T) ( = JCM 19513(T) = BCRC 80696(T)); a second strain of the same species is CC-AMHZ-5 ( = JCM 19512 = BCRC 80697). In addition, emended descriptions of the species Pseudomonas pohangensis, Pseudomonas benzenivorans and Pseudomonas segetis are also proposed.

Topics: Animals; Bacterial Typing Techniques; Birds; DNA, Bacterial; Fatty Acids; Feces; Molecular Sequence Data; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Polyamines; Pseudomonas; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Taiwan; Ubiquinone

2,3-Oxidosqualene is an intermediate in cholesterol biosynthesis and 2,3:22,23-dioxidosqualene act as the substrate for an alternative pathway that produces 24(S),25-epoxycholesterol which effects cholesterol homeostasis. In light of our previous findings concerning the biological effects of certain epoxidated all-trans-polyisoprenes, the effects of squalene carrying epoxy moieties on the second and third isoprene residues were investigated here. In cultures of HepG2 cells both monoepoxides of squalene and one of their hydrolytic products inhibited cholesterol synthesis and stimulated the synthesis of coenzyme Q (CoQ). Upon prolonged treatment the cholesterol content of these cells and its labeling with [(3)H]mevalonate were reduced, while the amount and labeling of CoQ increased. Injection of the squalene monoepoxides into mice once daily for 6days elevated the level of CoQ in their blood, but did not change the cholesterol level. The same effects were observed upon treatment of apoE-deficient mice and diabetic GK-rats. This treatment increased the hepatic level of CoQ10 in mice, but the amount of CoQ9, which is the major form, was unaffected. The presence of the active compounds in the blood was supported by the finding that cholesterol synthesis in the white blood cells was inhibited. Since the ratio of CoQ9/CoQ10 varies depending on the experimental conditions, the cells were titrated with substrate and inhibitors, leading to the conclusion that the intracellular isopentenyl-PP pool is a regulator of this ratio. Our present findings indicate that oxidosqualenes may be useful for stimulating both the synthesis and level of CoQ both in vitro and in vivo.

Topics: Animals; Bridged Bicyclo Compounds, Heterocyclic; Cholesterol; Diabetes Mellitus, Experimental; Etidronic Acid; Hemiterpenes; Hep G2 Cells; Humans; Lovastatin; Male; Mevalonic Acid; Mice; Mice, Inbred C57BL; Mice, Knockout; Organophosphorus Compounds; Rats; Rats, Wistar; Risedronic Acid; Squalene; Tricarboxylic Acids; Ubiquinone

Five acetic acid bacteria isolates, awK9_3, awK9_4 ( = LMG 27543), awK9_5 ( = LMG 28092), awK9_6 and awK9_9, obtained during a study of micro-organisms present in traditionally produced kefir, were grouped on the basis of their MALDI-TOF MS profile with LMG 1530 and LMG 1531(T), two strains currently classified as members of the genus Acetobacter. Phylogenetic analysis based on nearly complete 16S rRNA gene sequences as well as on concatenated partial sequences of the housekeeping genes dnaK, groEL and rpoB indicated that these isolates were representatives of a single novel species together with LMG 1530 and LMG 1531(T) in the genus Acetobacter, with Acetobacter aceti, Acetobacter nitrogenifigens, Acetobacter oeni and Acetobacter estunensis as nearest phylogenetic neighbours. Pairwise similarity of 16S rRNA gene sequences between LMG 1531(T) and the type strains of the above-mentioned species were 99.7%, 99.1%, 98.4% and 98.2%, respectively. DNA-DNA hybridizations confirmed that status, while amplified fragment length polymorphism (AFLP) and random amplified polymorphic DNA (RAPD) data indicated that LMG 1531(T), LMG 1530, LMG 27543 and LMG 28092 represent at least two different strains of the novel species. The major fatty acid of LMG 1531(T) and LMG 27543 was C18 : 1ω7c. The major ubiquinone present was Q-9 and the DNA G+C contents of LMG 1531(T) and LMG 27543 were 58.3 and 56.7 mol%, respectively. The strains were able to grow on D-fructose and D-sorbitol as a single carbon source. They were also able to grow on yeast extract with 30% D-glucose and on standard medium with pH 3.6 or containing 1% NaCl. They had a weak ability to produce acid from d-arabinose. These features enabled their differentiation from their nearest phylogenetic neighbours. The name Acetobacter sicerae sp. nov. is proposed with LMG 1531(T) ( = NCIMB 8941(T)) as the type strain.

Topics: Acetobacter; Alcoholic Beverages; Amplified Fragment Length Polymorphism Analysis; Bacterial Typing Techniques; Base Composition; Cultured Milk Products; DNA, Bacterial; Fatty Acids; Genes, Bacterial; Molecular Sequence Data; Nucleic Acid Hybridization; Phylogeny; Random Amplified Polymorphic DNA Technique; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone

A Gram-stain-negative, non-motile bacterial strain designated 1NM-4(T) was isolated from an abandoned lead-zinc ore mine site in Mei County, Meizhou, Guangdong Province, southern China. The isolate was light yellow, strictly aerobic, oxidase-negative and catalase-positive. Phylogenetic analyses based on 16S rRNA, rpoB and gyrB gene sequences, together with DNA-DNA hybridization values less than 70%, revealed that strain 1NM-4(T) belongs to the genus Acinetobacter and may represent a novel species. The major respiratory quinone was ubiquinone 9 (Q-9) and the major cellular fatty acids consisted of C18:1ω9c, summed feature 3 (C16:1ω7c and/or C16:1ω6c), C16:0 and C12:0. The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, an unidentified aminolipid and two unidentified phospholipids. The genomic DNA G+C content of strain 1NM-4(T) was 47.17 ± 0.02 mol%. Based on phenotypic, phylogenetic and chemotaxonomic characteristics, strain 1NM-4(T) should be assigned to a novel species of the genus Acinetobacter, for which the name Acinetobacter guangdongensis sp. nov. is proposed. The type strain is 1NM-4(T) ( = GIMCC 1.656(T) = CCTCC AB 2014199(T) = KCTC 42012(T)).

Topics: Acinetobacter; Bacterial Typing Techniques; Base Composition; China; DNA, Bacterial; Fatty Acids; Genes, Bacterial; Lead; Mining; Molecular Sequence Data; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Pigmentation; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Soil Microbiology; Ubiquinone; Zinc

A novel Gram-stain-negative, aerobic, non-endospore-forming, non-pigmented, rod-shaped, slightly halophilic bacterium, designated GBPy5(T), was isolated from aquatic plants of the Gomishan wetland, Iran. Cells of strain GBPy5(T) were motile. Growth occurred with between 1 and 10% (w/v) NaCl and the isolate grew optimally with 3% (w/v) NaCl. The optimum pH and temperature for growth of the strain were pH 8.0 and 30 °C, respectively, while it was able to grow over a pH range of 6.5-9.0 and a temperature range of 4-35 °C. Phylogenetic analysis, based on 16S rRNA gene sequences, revealed that strain GBPy5(T) is a member of the genus Pseudomonas forming a monophyletic branch. The novel strain exhibited 16S rRNA gene sequence similarity of 95.4% with type strains of Pseudomonas guariconensis PCAVU11(T) and Pseudomonas sabulinigri J64(T), respectively. The major cellular fatty acids of the isolate were C18:1ω7c (37.8%), C16:0 (14.9%), C16:1ω7c (12.9%), C12:0 3-OH (7.1%) and C12:0 (7.0%). The polar lipid pattern of strain GBPy5(T) comprised phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and one phospholipid. Ubiquinone 9 (Q-9) was the predominant lipoquinone. The G+C content of the genomic DNA of strain GBPy5(T) was 59.2 mol%. On the basis of the phenotypic and phylogenetic data, strain GBPY5(T) represents a novel species of the genus Pseudomonas, for which the name Pseudomonas salegens sp. nov. is proposed. The type strain is GBPy5(T) ( = IBRC-M 10762(T) = CECT 8338(T)).

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Iran; Molecular Sequence Data; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Pseudomonas; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone; Wetlands

A novel Gram-negative, obligate aerobic, non-motile, and both coccobacillus- and bacillus-shaped bacterium, designated strain HYN18(T), was isolated from the intestinal tract of a honey bee (Apis mellifera). The isolate was oxidase-negative and catalase-positive. Strain HYN18(T) showed optimum growth at 25°C, pH 6-7, and in the presence of 1% (w/v) NaCl in trypticase soy broth medium. The isolate was negative for hydrolyses of starch, casein, gelatin and urea, indole production from tryptone and hemolysis on sheep blood agar. A phylogenetic analysis based on the 16S rRNA gene and rpoB gene sequence showed that strain HYN18(T) was most closely related to Acinetobacter nectaris SAP 763.2(T) and A. boissieri SAP 284.1(T) with 98.3% and 98.1% similarity (16S rRNA gene), respectively, and 84.4% similarity with Acinetobacter nectaris SAP 763.2(T) (rpoB gene). The major cellular fatty acids were summed features 3 (comprising C16:1ω7c /C16:1ω6c ), C12:0 and C16:0. The main isoprenoid quinone was ubiquinone-9 (Q-9). The polar lipids of strain HYN18(T) were phosphatidylethanolamine, three unidentified lipids, an unidentified phospholipid and an unidentified glycolipid. The DNA G+C content was 40.6 mol%. DNA-DNA hybridization experiments indicated less than 33 ± 10% relatedness to the closest phylogenetic species, Acinetobacter nectaris SAP 763.2(T). Thus, the phenotypic, phylogenetic and genotypic analyses indicate that strain HYN18(T) is a novel species within the genus Acinetobacter, for which the name Acinetobacter apis is proposed. The type strain is HYN18(T) (=KACC 16906(T) =JCM 18575(T)).

Topics: Acinetobacter; Animals; Bacterial Typing Techniques; Base Composition; Bees; DNA, Bacterial; Fatty Acids; Genotype; Intestines; Molecular Sequence Data; Nucleic Acid Hybridization; Phenotype; Phosphatidylethanolamines; Phospholipids; Phylogeny; Quinones; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone

A taxonomic study was carried out on a novel aerobic bacterial strain, designated CC-CY503(T), isolated from food-waste compost in Taiwan. Cells were Gram-stain-negative short rods, motile by means of a monopolar flagellum. Strain CC-CY503(T) was able to grow at 20-50 °C and pH 6.0-10.0 and to tolerate <6% NaCl (w/v). Phylogenetic analysis of 16S rRNA gene sequences showed that this bacterium belonged to the genus Pseudomonas, with Pseudomonas pertucinogena ATCC 190(T) as the closest neighbour, sharing a sequence similarity of 97.9%. The DNA-DNA relatedness value of strain CC-CY503(T) with P. pertucinogena ATCC 190(T) was 37.8 ± 2.3%. The phylogenetic trees reconstructed based on gyrB and rpoB gene sequences supported the classification of strain CC-CY503(T) as a novel member of the genus Pseudomonas. The predominant quinone system was ubiquinone (Q-9) and the DNA G+C content was 63.1 ± 0.4 mol%. The major fatty acids were C(12:0), C(16:0), C(17:0) cyclo, C(19:0) cyclo ω8c and summed features 3 and 8 consisting of C(16:1)ω7c/C(16:1)ω6c and C(18:1)ω7c/C(18:1)ω6c, respectively. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and phosphatidylcholine. On the basis of its distinct phylogenetic, phenotypic and chemotaxonomic features, strain CC-CY503(T) ( =BCRC 80437(T) =JCM 18415(T)) is proposed to represent a novel species within the genus Pseudomonas, for which the name Pseudomonas formosensis sp. nov. is proposed.

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Food Microbiology; Molecular Sequence Data; Nucleic Acid Hybridization; Phospholipids; Phylogeny; Pseudomonas; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Soil Microbiology; Solid Waste; Taiwan; Ubiquinone

Membrane lipid composition is an important correlate of the rate of aging of animals. Dietary methionine restriction (MetR) increases lifespan in rodents. The underlying mechanisms have not been elucidated but could include changes in tissue lipidomes. In this work, we demonstrate that 80% MetR in mice induces marked changes in the brain, spinal cord, and liver lipidomes. Further, at least 50% of the lipids changed are common in the brain and spinal cord but not in the liver, suggesting a nervous system-specific lipidomic profile of MetR. The differentially expressed lipids includes (a) specific phospholipid species, which could reflect adaptive membrane responses, (b) sphingolipids, which could lead to changes in ceramide signaling pathways, and (c) the physiologically redox-relevant ubiquinone 9, indicating adaptations in phase II antioxidant response metabolism. In addition, specific oxidation products derived from cholesterol, phosphatidylcholine, and phosphatidylethanolamine were significantly decreased in the brain, spinal cord, and liver from MetR mice. These results demonstrate the importance of adaptive responses of membrane lipids leading to increased stress resistance as a major mechanistic contributor to the lowered rate of aging in MetR mice.

Topics: Adaptation, Physiological; Aging; Animals; Brain; Cholesterol; Female; Lipid Metabolism; Liver; Longevity; Male; Methionine; Mice; Mice, Inbred C57BL; Oxidation-Reduction; Oxidative Stress; Phosphatidylcholines; Phosphatidylethanolamines; Reactive Oxygen Species; Spinal Cord; Ubiquinone

The interaction of dietary fats and carbohydrates on liver mitochondria were examined in male FBNF1 rats fed 20 different low-fat isocaloric diets. Animal growth rates and mitochondrial respiratory parameters were essentially unaffected, but mass spectrometry-based mitochondrial lipidomics profiling revealed increased levels of cardiolipins (CLs), a family of phospholipids essential for mitochondrial structure and function, in rats fed saturated or trans fat-based diets with a high glycemic index. These mitochondria showed elevated monolysocardiolipins (a CL precursor/product of CL degradation), elevated ratio of trans-phosphocholine (PC) (18:1/18:1) to cis-PC (18:1/18:1) (a marker of thiyl radical stress), and decreased ubiquinone Q9; the latter two of which imply a low-grade mitochondrial redox abnormality. Extended analysis demonstrated: i) dietary fats and, to a lesser extent, carbohydrates induce changes in the relative abundance of specific CL species; ii) fatty acid (FA) incorporation into mature CLs undergoes both positive (>400-fold) and negative (2.5-fold) regulation; and iii) dietary lipid abundance and incorporation of FAs into both the CL pool and specific mature tetra-acyl CLs are inversely related, suggesting previously unobserved compensatory regulation. This study reveals previously unobserved complexity/regulation of the central lipid in mitochondrial metabolism.

Topics: Animals; Cardiolipins; Cell Respiration; Diet; Fatty Acids, Omega-3; Fatty Acids, Omega-6; Glycemic Index; Liver; Male; Mitochondria, Liver; Oxidative Stress; Oxygen Consumption; Rats; Ubiquinone

We examined the effect of vitamin E depletion on liver oxidative damage in rats with water-immersion restraint stress (WIRS). Male Wistar rats were fed a normal diet (N) or vitamin E-depleted diet (VE-D) for 4 wk. N- and VE-D-fed rats were exposed to WIRS for 6 h. The activities of serum transaminases and lactate dehydrogenase and serum ascorbic acid concentration were similar in both diet groups. WIRS exposure increased these serum enzyme activities and the serum ascorbic acid concentration in both diet groups but the ratios of these increases were higher in VE-D-fed rats than in N-fed rats. Serum and liver α-tocopherol concentrations in VE-D-rats were approximately 50% and 30% of those in N-fed rats, respectively. WIRS exposure reduced liver α-tocopherol concentration in VE-D-fed rats, but not in N-fed rats. Liver ascorbic acid and reduced glutathione concentrations were higher in the VE-D-fed group than in the N-fed group. WIRS exposure reduced liver ascorbic acid and reduced glutathione concentrations in both diet groups. There were no differences in liver concentrations of coenzyme Q9 or coenzyme Q10 in the reduced form between the N- and VE-D-fed groups. WIRS exposure reduced liver concentrations of coenzyme Q9 and coenzyme Q10 in the reduced form in both diet groups. Liver lipid peroxide concentration was higher in the VE-D-fed group than in the N-fed group. WIRS exposure raised liver lipid peroxide concentration more in the VE-D-fed group than in the N-fed group. These results indicate that vitamin E depletion enhances liver oxidative damage in rats with WIRS.

Topics: Animals; Ascorbic Acid; Diet; Glutathione; L-Lactate Dehydrogenase; Lipid Peroxides; Liver; Male; Oxidative Stress; Rats; Rats, Wistar; Restraint, Physical; Stress, Physiological; Transaminases; Ubiquinone; Vitamin E

A Gram-stain-negative, aerobic, rod-like, motile by peritrichous flagella and moderately halophilic bacterium, designated strain B6(T), was isolated a deep-sea sediment collected from the South Atlantic Ocean. The isolate grew with 0.5-15 % (w/v) NaCl, at 4-37 °C and pH 5.0-8.5 and showed a high tolerance to zinc, manganese, cobalt and copper ions. The major fatty acids were C16 : 0, C19 : 0 cyclo ω8c, C12 : 0 3-OH and C12 : 0. The predominant ubiquinone was Q-9. The genomic DNA G+C content was 61.1 mol%. Phylogenetic analysis based on 16S rRNA gene comparisons indicated that strain B6(T) belonged to the genus Halomonas, and the closest relative was Halomonas xinjiangensis TRM 0175(T) (96.1 %). Based upon the phenotypic, chemotaxonomic and genetic data, strain B6(T) represents a novel species from the genus Halomonas, for which the name Halomonas zincidurans sp. nov. is proposed. The type strain is B6(T) ( = CGMCC 1.12450(T) = JCM 18472(T)).

Topics: Atlantic Ocean; Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Geologic Sediments; Halomonas; Metals, Heavy; Molecular Sequence Data; Phylogeny; RNA, Ribosomal, 16S; Seawater; Ubiquinone

Two Gram-negative, facultatively anaerobic, rod-shaped bacteria, strains EF212(T) and PS125(T), were isolated from the octocorals Eunicea fusca and Plexaura sp., respectively. EF212(T) was isolated from a specimen of E. fusca collected off the coast of Florida, USA, and PS125(T) was isolated from a specimen of Plexaura sp. collected off the coast of Bimini, Bahamas. Analysis of the nearly full-length 16S rRNA gene sequences showed that these novel strains were most closely related to Endozoicomonas montiporae CL-33(T), E. elysicola MKT110(T) and E. numazuensis HC50(T) (EF212(T), 95.6-97.2 % identity; PS125(T), 95.1-96.4 % identity). DNA-DNA hybridization values among EF212(T), PS125(T), E. montiporae LMG 24815(T) and E. elysicola KCTC 12372(T) were far below the 70 % cut-off, with all values for duplicate measurements being less than 35 %. Both EF212(T) and PS125(T) required NaCl for growth and showed optimal growth at 2-3 % NaCl, 22-30 °C and pH 8.0. The predominant cellular fatty acids were summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c), summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c), C16 : 0 and C14 : 0. The DNA G+C content of EF212(T) was 48.6 mol% and that of PS125(T) was 47.5 mol%. In addition to the genotypic differences observed between the two novel strains and related type strains, phenotypic and chemotaxonomic experiments also revealed differences between strains. Thus, strains EF212(T) and PS125(T) represent novel species of the genus Endozoicomonas, for which the names Endozoicomonas euniceicola sp. nov. and Endozoicomonas gorgoniicola sp. nov., respectively, are proposed. The type strains are EF212(T) ( = NCCB 100458(T) = DSM 26535(T)) for Endozoicomonas euniceicola sp. nov. and PS125(T) ( = NCCB 100438(T) = CECT 8353(T)) for Endozoicomonas gorgoniicola sp. nov. An emended description of the genus Endozoicomonas is also provided to encompass differences observed in the results of genotypic, chemotaxonomic and phenotypic tests compared from the original and amended genus descriptions.

Topics: Animals; Anthozoa; Bacterial Typing Techniques; Bahamas; Base Composition; DNA, Bacterial; Fatty Acids; Florida; Gammaproteobacteria; Genotype; Molecular Sequence Data; Nucleic Acid Hybridization; Phylogeny; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone

An aerobic, Gram-stain-negative, rod-shaped bacterium (designated strain CC-G9A(T)), motile by a polar-flagellum, was isolated from a hot spring water sample in Taiwan. Strain CC-G9A(T) could grow at 20-42 °C, pH 6.0-10.0 and tolerate up to 7% (w/v) NaCl. The 16S rRNA gene sequence analysis of strain CC-G9A(T) showed pairwise sequence similarity to Pseudomonas mendocina LMG 1223(T) (97.7%), Pseudomonas alcaligenes ATCC 14909(T) (97.8 %), Pseudomonas alcaliphila DSM 17744(T) (97.8 %), Pseudomonas toyotomiensis JCM 15604(T) (97.6 %), Pseudomonas oleovorans subsp. lubricantis DSM 21016(T) (97.6 %) and Pseudomonas argentinensis BCRC 17807(T) (97.5 %), and lower sequence similarity to other species of the genus Pseudomonas. According to DNA-DNA association analysis, the relatedness of strain CC-G9A(T) to P. mendocina BCRC 10458(T), P. alcaliphila DSM 17744(T), P. alcaligenes BCRC 11893(T), P. oleovorans subsp. lubricantis DSM 21016(T), P. argentinensis BCRC 17807(T) and P. oleovorans subsp. oleovorans BCRC 11902 was 55.1±3.1, 13.7±1.5, 14.1±1.8, 58.5±1.1, 28.9±2.0 and 28.6±1.8 %, respectively. The evolutionary trees reconstructed based on 16S rRNA, gyrB and rpoB gene sequences revealed varying phylogenetic neighbourhoods of strain CC-G9A(T) with regard to the most closely related type strains. The predominant quinone system was ubiquinone (Q-9) and the DNA G+C content was 64.3±1.3 mol%. The major fatty acids were C10 : 0 3-OH, C12 : 0, C12 : 0 3-OH, C16 : 0 and summed features 3 and 8 consisting of C16 : 1ω7c/C16 : 1ω6c and C18 : 1ω7c/C18 : 1ω6c, respectively. The major polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine and phosphatidylglycerol. According to distinct phylogenetic, phenotypic and chemotaxonomic features, strain CC-G9A(T) is proposed to represent a novel species within the genus Pseudomonas for which the name Pseudomonas guguanensis sp. nov. is proposed. The type strain is CC-G9A(T) ( = BCRC 80438(T) = JCM 18416(T)).

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Genes, Bacterial; Hot Springs; Molecular Sequence Data; Nucleic Acid Hybridization; Phylogeny; Pseudomonas; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Taiwan; Ubiquinone

A Gram-negative, straight to slightly curved rod-shaped bacterium, motile with peritrichous flagella, designated SgZ-6(T), was isolated from an electroactive biofilm and was characterized by means of a polyphasic approach. Growth occurred with 0-5.0 % (w/v) NaCl (optimum 1 %), at pH 6.0-10.0 (optimum pH 7.0) and at 10-42 °C (optimum 30 °C) in trypticase soya broth. Phylogenetic analyses based on the 16S rRNA and gyrB genes identified the isolate as a member of a novel species of the genus Pseudomonas. Strain SgZ-6(T) exhibited the highest 16S rRNA gene sequence similarity to 'Pseudomonas linyingensis' CGMCC 1.10701 (97.5 %), followed by Pseudomonas sagittaria JCM 18195(T) (97.4 %), P. oleovorans subsp. lubricantis DSM 21016(T) (96.6 %), P. tuomuerensis JCM 14085(T) (96.5 %) and P. alcaliphila JCM 10630(T) (96.4 %). Strain SgZ-6(T) showed the highest gyrB gene sequence similarity of 93.7 % to 'P. linyingensis' CGMCC 1.10701 among all type strains of genus Pseudomonas. DNA-DNA pairing studies showed that strain SgZ-6(T) displayed 47.1 and 40.3 % relatedness to 'P. linyingensis' CGMCC 1.10701 and P. sagittaria JCM 18195(T), respectively. The major isoprenoid quinone was ubiquinone 9 (Q-9). The whole-cell fatty acids consisted mainly of summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c), C16 : 0 and summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c). The DNA G+C content of the genomic DNA was 68.1 mol%. On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain SgZ-6(T) is proposed to represent a novel species of the genus Pseudomonas, for which the name Pseudomonas guangdongensis sp. nov. is proposed. The type strain is SgZ-6(T) ( = CCTCC AB 2012022(T) = KACC 16606(T)). An emended description of the genus Pseudomonas is also proposed.

Topics: Bacterial Typing Techniques; Base Composition; Bioelectric Energy Sources; Biofilms; DNA, Bacterial; Fatty Acids; Molecular Sequence Data; Nucleic Acid Hybridization; Phylogeny; Pseudomonas; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ubiquinone

Four Gram-negative, moderately halophilic, exopolysaccharide-producing strains, designated AAD6(T), AAD4, AAD17 and AAD21, were isolated from Çamaltı Saltern Area, a wildlife reserve in Sasalı, İzmir province located in the Aegean Region of Turkey. The isolates grew at an optimum NaCl concentration of 10% (w/v). The major cellular fatty acids were C(16:0), C(18:1)ω7c, C(16:1)ω7c and C(12:0) 3OH, respectively and the predominant lipoquinone was ubiquinone Q-9. The G+C content of the genomic DNA of strains AAD6(T), AAD4, AAD17 and AAD21 was 63.0, 63.3, 62.8 and 62.6 mol %, respectively. Comparative 16S rRNA gene sequence studies showed that the isolates belonged to the genus Halomonas. The DNA-DNA hybridization mean values between the representative strain AAD6(T) and the closely related species Halomonas salina DSM 5928(T), Halomonas halophila DSM 4770(T), Halomonas maura DSM 13445(T), Halomonas organivorans DSM 16226(T), Halomonas elongata DSM 2581(T), Halomonas koreensis JCM 12237(T) and Halomonas nitroreducens LMG 24185, were 40.8, 39.6, 24.2, 23.3, 12.6, 14.5 and 12.2%, respectively. Based on these data the strains represent a novel species of the genus Halomonas for which the name Halomonas smyrnensis sp. nov. is proposed. The type strain is AAD6(T) (= DSM 21644(T) = JCM 15723(T)).

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Halomonas; Molecular Sequence Data; Nucleic Acid Hybridization; Phylogeny; Polysaccharides, Bacterial; Ponds; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Soil Microbiology; Turkey; Ubiquinone

Four orange-pigmented isolates, L7-456, L7-484(T), L9-479 and L9-753(T), originating from surface-sterilized leaf tissues of Jatropha curcas L. cultivars were characterized using a polyphasic taxonomic approach. Phylogenetic analyses based on 16S rRNA gene sequences indicated that all four isolates belong to the genus Aureimonas. In these analyses, strain L7-484(T) appeared to be most closely related to Aureimonas ureilytica 5715S-12(T) (95.7 % sequence identity). The 16S rRNA gene sequences of strains L7-456, L9-479 and L9-753(T) were found to be identical and also shared the highest similarity with A. ureilytica 5715S-12(T) (97.5 %). Both L7-484(T) and L9-753(T) contained Q-10 and Q-9 as predominant ubiquinones and diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, phosphatidylmonomethylethanolamine, phosphatidylethanolamine, phosphatidyldimethylethanolamine, sulfoquinovosyldiacylglycerol and an aminophospholipid as the major polar lipids. C18 : 1ω7c and C16 : 0 were the major fatty acids. Similar to other species in the genus Aureimonas, hydroxylated fatty acids (e.g. C18 : 1 2-OH) and cyclic fatty acids (C19 : 0 cyclo ω8c) were also present. The DNA G+C contents of L7-484(T) and L9-753(T) were 66.1 and 69.4 mol%, respectively. Strains L7-484(T) and L9-753(T) exhibited less than 40 % DNA-DNA hybridization both between themselves and to A. ureilytica KACC 11607(T). Our results support the proposal that strain L7-484(T) represents a novel species within the genus Aureimonas, for which the name Aureimonas jatrophae sp. nov. is proposed, and that strains L9-753(T), L7-456 ( = KACC 16229  = DSM 25023) and L9-479 ( = KACC 16228  = DSM 25024) represent a second novel species within the genus, for which the name Aureimonas phyllosphaerae sp. nov. is proposed. The type strains of Aureimonas jatrophae sp. nov. and Aureimonas phyllosphaerae sp. nov. are respectively L7-484(T) ( = KACC 16230(T)  = DSM 25025(T)) and L9-753(T) ( = KACC 16231(T)  = DSM 25026(T)).

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Flavobacteriaceae; Jatropha; Microscopy, Electron, Transmission; Molecular Sequence Data; Nucleic Acid Hybridization; Phylogeny; Plant Leaves; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Singapore; Ubiquinone

A Gram-negative, aerobic, non-motile, non-spore-forming, cocci-shaped, red-pigmented bacterium, designated strain PB-K8(T), was isolated from grass soil sampled in Daejeon, Republic of Korea. Comparative 16S rRNA gene sequence studies showed that the isolate was clearly affiliated with the class Alphaproteobacteria, and was most closely related to Belnapia moabensis DSM 16746(T) and Belnapia rosea DSM 23312(T), showing a 16S rRNA gene sequence similarity to the type strains of each species of 98.4 and 97.2%, respectively. The cells of strain PB-K8(T) formed red colonies on R2A agar, contained Q-9 as the predominant ubiquinone and included summed feature 3 (C16:1ω7c C16:1ω6c), C16:0, summed feature 8 (C18:1ω7c/C18:1ω6c), C18:1 2-OH and C19:0 cyclo ω8c as the major fatty acids. The G+C content of the genomic DNA of strain PB-K8(T) was 72.1 mol%. Thus, the combined genotypic and phenotypic data supported the conclusion that strain PB-K8(T) represents a novel species of the genus Belnapia, for which the name Belnapia soli sp. nov. is proposed. The type strain is PB-K8(T) (=KCTC 23765(T)=JCM 18033(T)).

Topics: Acetobacteraceae; Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Molecular Sequence Data; Nucleic Acid Hybridization; Phylogeny; Poaceae; Republic of Korea; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Soil Microbiology; Ubiquinone

Primary human CoQ(10) deficiencies are clinically heterogeneous diseases caused by mutations in PDSS2 and other genes required for CoQ(10) biosynthesis. Our in vitro studies of PDSS2 mutant fibroblasts, with <20% CoQ(10) of control cells, revealed reduced activity of CoQ(10)-dependent complex II+III and ATP synthesis, without amplification of reactive oxygen species (ROS), markers of oxidative damage, or antioxidant defenses. In contrast, COQ2 and ADCK3 mutant fibroblasts, with 30-50% CoQ(10) of controls, showed milder bioenergetic defects but significantly increased ROS and oxidation of lipids and proteins. We hypothesized that absence of oxidative stress markers and cell death in PDSS2 mutant fibroblasts were due to the extreme severity of CoQ(10) deficiency. Here, we have investigated in vivo effects of Pdss2 deficiency in affected and unaffected organs of CBA/Pdss2(kd/kd) mice at presymptomatic, phenotypic-onset, and end-stages of the disease. Although Pdss2 mutant mice manifest widespread CoQ(9) deficiency and mitochondrial respiratory chain abnormalities, only affected organs show increased ROS production, oxidative stress, mitochondrial DNA depletion, and reduced citrate synthase activity, an index of mitochondrial mass. Our data indicate that kidney-specific loss of mitochondria triggered by oxidative stress may be the cause of renal failure in Pdss2(kd/kd) mice.

Topics: Adenosine Diphosphate; Adenosine Triphosphate; Alkyl and Aryl Transferases; Animals; DNA, Mitochondrial; Electron Transport; Fibroblasts; Humans; Kidney; Mice; Mice, Inbred CBA; Mice, Mutant Strains; Mitochondria; Oxidative Stress; Tissue Distribution; Ubiquinone

An aerobic, Gram-stain-negative, rod-shaped bacterium with a single polar flagellum, designated CC-OPY-1(T), was isolated from an oil-contaminated site in Taiwan. CC-OPY-1(T) produces siderophores, and can grow at temperatures of 25-37 °C and pH 5.0-9.0 and tolerate <5 % (w/v) NaCl. The 16S rRNA gene sequence analysis of CC-OPY-1(T) showed high pairwise sequence similarity to Pseudomonas alcaligenes BCRC 11893(T) (97.1 %), Pseudomonas. alcaliphila DSM 17744(T) (97.1 %), Pseudomonas tuomuerensis JCM 14085(T) (97.1 %), Pseudomonas toyotomiensis JCM 15604(T) (96.9 %) and lower sequence similarity to remaining species of the genus Pseudomonas. The phylogenetic trees reconstructed based on gyrB and rpoB gene sequences supported the classification of CC-OPY-1(T) as a novel member of the genus Pseudomonas. The predominant quinone system of strain CC-OPY-1T was ubiquinone (Q-9) and the DNA G+C content was 68.4 ± 0.3 mol%. The major fatty acids were C12 : 0, C16 : 0, C17 : 0 cyclo and summed features 3 and 8 consisting of C16 : 1ω7c/C16 : 1ω6c and C18 : 1ω7c/C18 : 1ω6c, respectively. The major polar lipids were phosphatidylethanolamine (PE), phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), phosphatidylcholine (PC) and two unknown phospholipids (PL1-2). Due to distinct phylogenetic, phenotypic and chemotaxonomic features, CC-OPY-1(T) is proposed to represent a novel species within the genus Pseudomonas for which the name Pseudomonas sagittaria sp. nov. is proposed. The type strain is CC-OPY-1(T) ( = BCRC 80399(T) = JCM 18195(T)).

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Molecular Sequence Data; Nucleic Acid Hybridization; Petroleum Pollution; Phospholipids; Phylogeny; Pseudomonas; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Soil Microbiology; Soil Pollutants; Taiwan; Ubiquinone

Coenzyme Q10 (CoQ(10)) or ubiquinone is a well-known component of the mitochondrial respiratory chain. In humans, CoQ(10) deficiency causes a mitochondrial syndrome with an unexplained variability in the clinical presentations. To try to understand this heterogeneity in the clinical phenotypes, we have generated a Coq9 Knockin (R239X) mouse model. The lack of a functional Coq9 protein in homozygous Coq9 mutant (Coq9(X/X)) mice causes a severe reduction in the Coq7 protein and, as consequence, a widespread CoQ deficiency and accumulation of demethoxyubiquinone. The deficit in CoQ induces a brain-specific impairment of mitochondrial bioenergetics performance, a reduction in respiratory control ratio, ATP levels and ATP/ADP ratio and specific loss of respiratory complex I. These effects lead to neuronal death and demyelinization with severe vacuolization and astrogliosis in the brain of Coq9(X/X) mice that consequently die between 3 and 6 months of age. These results suggest that the instability of mitochondrial complex I in the brain, as a primary event, triggers the development of mitochondrial encephalomyopathy associated with CoQ deficiency.

Topics: Adenosine Diphosphate; Adenosine Triphosphate; Animals; Brain; Electron Transport Complex I; Female; Humans; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Mitochondrial Encephalomyopathies; Ubiquinone

To examine the hypothesis that resuscitative hypothermia would (1) reduce fluid requirements and reactive oxygen species production during a period of resuscitation and (2) improve survival after hemorrhagic shock (HS) in rats.. Sixteen rats underwent an HS phase (phase I: 0-75 minutes), with pressure-controlled HS at a mean arterial pressure of 30 mm Hg ± 5 mm Hg; a resuscitation phase (phase II: 75-150 minutes), with fluid resuscitation to maintain mean arterial pressure ≥75 mm Hg; and an observation phase (phase III: from 150 minutes to 72 hours). During phase II, eight rats were randomized into a normothermia group (group 1: 38°C) or a hypothermia group (group 2: 34°C). Fluid requirements during phase II and survival at 72 hours were compared between groups. Plasma levels of Vitamin E and %coenzyme Q9 (%CoQ9) were also assessed.. The fluid requirement during resuscitation in phase II was 8.2 ± 1.4 mL/100 g in group 1 versus 2.1 mL/100 g ± 0.7 mL/100 g in group 2 (p < 0.01). Vitamin E level decreased to 10.8 μmol/L ± 1.8 μmol/L during HS in all rats. After resuscitation, it was restored to a baseline level of 15.9 μmol/L ± 3.1 μmol/L in group 2 but remained at 10.2 μmol/L ± 0.8 μmol/L in group 1 (p < 0.05). %CoQ9 did not differ significantly between the groups. At 72 hours, six of eight rats in group 1, and all rats in group 2 survived (NS).. In a rat HS model, hypothermia during resuscitation from HS reduces resuscitation fluid volume required to maintain blood pressure and restores Vitamin E to the baseline level, and appears to have no adverse impact on long survival after HS.

Topics: Animals; Blood Pressure; Disease Models, Animal; Fluid Therapy; Hemodynamics; Hypothermia, Induced; Male; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Respiratory Rate; Resuscitation; Shock, Hemorrhagic; Ubiquinone; Vitamin E

Ubiquinone (coenzyme Q) is the generic name of a class of lipid-soluble electron carriers formed of a redox active benzoquinone ring attached to a prenyl side chain. The length of the latter varies among species, and depends upon the product specificity of a trans-long-chain prenyl diphosphate synthase that elongates an allylic diphosphate precursor. In Arabidopsis, this enzyme is assumed to correspond to an endoplasmic reticulum-located solanesyl diphosphate synthase, although direct genetic evidence was lacking. In this study, the reconstruction of the functional network of Arabidopsis genes linked to ubiquinone biosynthesis singled out an unsuspected solanesyl diphosphate synthase candidate--product of gene At2g34630--that, extraordinarily, had been shown previously to be targeted to plastids and to contribute to the biosynthesis of gibberellins. Green fluorescent protein (GFP) fusion experiments in tobacco and Arabidopsis, and complementation of a yeast coq1 knockout lacking mitochondrial hexaprenyl diphosphate synthase demonstrated that At2g34630 is also targeted to mitochondria. At2g34630 is the main--if not sole--contributor to solanesyl diphosphate synthase activity required for the biosynthesis of ubiquinone, as demonstrated by the dramatic (75-80%) reduction of the ubiquinone pool size in corresponding RNAi lines. Overexpression of At2g34630 gave up to a 40% increase in ubiquinone content compared to wild-type plants. None of the silenced or overexpressing lines, in contrast, displayed altered levels of plastoquinone. Phylogenetic analyses revealed that At2g34630 is the only Arabidopsis trans-long-chain prenyl diphosphate synthase that clusters with the Coq1 orthologs involved in the biosynthesis of ubiquinone in other eukaryotes.

Topics: Alkyl and Aryl Transferases; Arabidopsis; Chloroplasts; Cloning, Molecular; Gene Knockout Techniques; Gene Regulatory Networks; Genetic Complementation Test; Green Fluorescent Proteins; Mitochondria; Mutation; Phylogeny; Plants, Genetically Modified; Plastoquinone; RNA Interference; Saccharomyces cerevisiae; Terpenes; Ubiquinone

Five yeast strains isolated from plant leaves collected in south-east Tibet formed cream to brownish colonies and produced asymmetrical ballistoconidia and CoQ-9 as the major ubiquinone. Sequence analysis of the 26S rRNA D1/D2 domain and the internal transcribed spacer region indicated that these strains represented two novel species of the genus Bensingtonia. The names Bensingtonia rectispora sp. nov. (type strain XZ 4C5(T) = CGMCC 2.02635(T) = CBS 10710(T)) and Bensingtonia bomiensis sp. nov. (type strain XZ 33D1(T) = CGMCC 2.02670(T) = CBS 10713(T)) are proposed for the two novel species, which are phylogenetically closely related to Bensingtonia naganoensis, Bensingtonia pseudonaganoensis and the type species of the genus, Bensingtonia ciliata.

Topics: Basidiomycota; DNA, Fungal; DNA, Ribosomal Spacer; Molecular Sequence Data; Mycological Typing Techniques; Phylogeny; Plant Leaves; RNA, Ribosomal; Sequence Analysis, DNA; Tibet; Ubiquinone

The redox status of mitochondrial coenzyme Q (CoQ) is an important marker for oxidative stress associated with several disorders such as Parkinson disease and Alzheimer disease. Altered redox status may be present in mitochondrial electron transport complex disorders. Intracellular CoQ levels reflect the functional status of the mitochondrial electron transport complex better than plasma levels. Here, we describe the method to determine the reduced and oxidized form of CoQ in white blood cells using LC-MS/MS.

Topics: Blood Chemical Analysis; Chromatography, High Pressure Liquid; Leukocytes; Oxidation-Reduction; Tandem Mass Spectrometry; Ubiquinone

A Gram-negative, motile, facultatively anaerobic rod, designated A36(T), was isolated from a dead ark clam found on the south coast of Korea. The isolate was catalase- and oxidase-negative. 16S rRNA gene sequence analysis indicated that strain A36(T) was most closely related to Kistimonas asteriae KMD 001(T), with which it shared 98.2% 16S rRNA gene sequence similarity. Strain A36(T) grew optimally at 30-37 °C, with 1% (w/v) NaCl and at pH 8.0. The major respiratory quinone was ubiquinone-9 (Q-9). The major polar lipids were phosphatidylserine, phosphoethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The major fatty acids were summed feature 3 (comprising C(16:1)ω7c and/or iso-C(15) 2-OH) and C(16:0). The genomic DNA G+C content was 47.3 mol%. DNA-DNA relatedness between the isolate and K. asteriae JCM 15607(T) was <25 ± 3%. Strain A36(T) represents a novel species of the genus Kistimonas, for which the name Kistimonas scapharcae sp. nov. is proposed. The type strain is A36(T) ( = KACC 16204(T)  = JCM 17805(T)). An emended description of the genus Kistimonas is also provided.

Topics: Animals; Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Gammaproteobacteria; Molecular Sequence Data; Phylogeny; Republic of Korea; RNA, Ribosomal, 16S; Scapharca; Sequence Analysis, DNA; Ubiquinone

The taxonomic position of a Gram-negative, non-motile, oxidase negative and catalase positive strain, A648(T), isolated from a hexachlorocyclohexane (HCH) dump site located in Lucknow, India, was ascertained by using a polyphasic approach. A comparative analysis of a partial sequence of the rpoB gene and the 16S rRNA gene sequence revealed that strain A648(T) belonged to the genus Acinetobacter. DNA-DNA relatedness values between strain A648(T) and other closely related members (16S rRNA gene sequence similarity greater than 97%), namely Acinetobacter radioresistens DSM 6976(T), A. venetianus ATCC 31012(T), A. baumannii LMG 1041(T), A. parvus LMG 21765(T) A. junii LMG 998(T) and A. soli JCM 15062(T), were found to be less than 8%. The major cellular fatty acids of strain A648(T) were 18:1ω9c (19.6%), summed feature 3 (15.9%), 16:0 (10.6%) and 12:0 (6.4%). The DNA G+C content was 40.4 mol%. The polar lipid profile of strain A648(T) indicated the presence of diphosphatidylglycerol, phosphatidylethanolamine, followed by phosphatidylglycerol and phosphatidylcholine. The predominant polyamine of strain A648(T) was 1,3-diaminopropane and moderate amounts of putrescine, spermidine and spermine were also detected. The respiratory quinone consisted of ubiquinone with nine isoprene units (Q-9). On the basis of DNA-DNA hybridization, phenotypic characteristics and chemotaxonomic and phylogenetic comparisons with other members of the genus Acinetobacter, strain A648(T) is found to be a novel species of the genus Acinetobacter, for which the name Acinetobacter indicus sp. nov. is proposed. The type strain is A648(T) ( = DSM 25388(T) = CCM 7832(T)).

Topics: Acinetobacter; Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Hexachlorocyclohexane; India; Molecular Sequence Data; Nucleic Acid Hybridization; Phylogeny; Polyamines; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Soil Microbiology; Soil Pollutants; Ubiquinone

A moderately halophilic bacterium (strain RS-16(T)) was isolated from saline soil in Rambla Salada, a Mediterranean hypersaline rambla in Murcia, south-east Spain. Cells of strain RS-16(T) were Gram-negative rods, oxidase-negative and motile by peritrichous flagella. Strain RS-16(T) required NaCl for growth, and grew between 1% and 30% (w/v) NaCl (optimum, 5-7.5%), at temperatures of between 4 °C and 41 °C (optimum, 32-37 °C), and at pH values of between 5 and 10 (optimum, pH 7). Strain RS-16(T) was chemo-organotrophic and its metabolism was respiratory with oxygen and nitrate as terminal electron acceptors. It produced acids from d-glucose and myo-inositol, accumulated poly-β-hydroxyalkanoate granules and produced cream colonies on MY 7.5% (w/v). The DNA G+C content of strain RS-16(T) was 56.2 mol%. A comparison of 16S rRNA gene sequences confirmed the relationship of strain RS-16(T) to species of the genus Halomonas. The most phylogenetically related species was Halomonas cerina SP4(T) (97.4%16S rRNA gene sequence similarity). In DNA-DNA hybridization assays strain RS-16(T) showed DNA-DNA relatedness values of 62.7 ± 3.09%, 64.5 ± 1.97% and 64.7 ± 1.74% to Halomonas cerina CECT 7282(T), Halomonas cerina CECT 7284 and Halomonas cerina CECT 7283, respectively. The major fatty acids of strain RS-16(T) were C(18:1)ω7c and C(16:0), and the predominant respiratory lipoquinone was ubiquinone, with nine isoprene units (Q-9). On the basis of these data, strain RS-16(T) is considered to represent a novel species of the genus Halomonas, for which the name Halomonas ramblicola sp. nov. is proposed. The type strain is RS-16(T) ( = CECT 7896(T) = LMG 26647(T)).

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Halomonas; Molecular Sequence Data; Nucleic Acid Hybridization; Phylogeny; RNA, Ribosomal, 16S; Salinity; Sequence Analysis, DNA; Soil Microbiology; Spain; Ubiquinone

Oxidative stress is believed to be a major contributory factor in the development of non alcoholic fatty liver disease (NAFLD), the most common liver disorder worldwide. In this study, the effects of high fat diet-induced NAFLD on Coenzyme Q (CoQ) metabolism and plasma oxidative stress markers in rats were investigated. Rats were fed a standard low fat diet (control) or a high fat diet (57% metabolizable energy as fat) for 18 weeks. The concentrations of total (reduced + oxidized) CoQ9 were increased by >2 fold in the plasma of animals fed the high fat diet, while those of total CoQ10 were unchanged. Reduced CoQ levels were raised, but oxidized CoQ levels were not, thus the proportion in the reduced form was increased by about 75%. A higher percentage of plasma CoQ9 as compared to CoQ10 was in the reduced form in both control and high fat fed rats. Plasma protein thiol (SH) levels were decreased in the high fat-fed rats as compared to the control group, but concentrations of lipid hydroperoxides and low density lipoprotein (LDL) conjugated dienes were unchanged. These results indicate that high fat diet-induced NAFLD in rats is associated with altered CoQ metabolism and increased protein, but not lipid, oxidative stress.

Topics: Animals; Dietary Fats; Lipoproteins, LDL; Male; Non-alcoholic Fatty Liver Disease; Oxidation-Reduction; Oxidative Stress; Rats; Rats, Wistar; Sulfhydryl Compounds; Ubiquinone

Coenzyme Q (ubiquinone or Q) is an essential lipid component of the mitochondrial electron transport chain. In Caenorhabditis elegans Q biosynthesis involves at least nine steps, including the hydroxylation of the hydroquinone ring by CLK-1 and two O-methylation steps mediated by COQ-3. We characterize two C. elegans coq-3 deletion mutants, and show that while each has defects in Q synthesis, their phenotypes are distinct. First generation homozygous coq-3(ok506) mutants are fertile when fed the standard lab diet of Q-replete OP50 Escherichia coli, but their second generation homozygous progeny does not reproduce. In contrast, the coq-3(qm188) deletion mutant remains sterile when fed Q-replete OP50. Quantitative PCR analyses suggest that the longer qm188 deletion may alter expression of the flanking nuo-3 and gdi-1 genes, located 5' and 3', respectively of coq-3 within an operon. We surmise that variable expression of nuo-3, a subunit of complex I, or of gdi-1, a guanine nucleotide dissociation inhibitor, may act in combination with defects in Q biosynthesis to produce a more severe phenotype. The phenotypes of both coq-3 mutants are more drastic as compared to the C. elegans clk-1 mutants. When fed OP50, clk-1 mutants reproduce for many generations, but show reduced fertility, slow behaviors, and enhanced life span. The coq-3 and clk-1 mutants all show arrested development and are sterile when fed the Q-deficient E. coli strain GD1 (harboring a mutation in the ubiG gene). However, unlike clk-1 mutant worms, neither coq-3 mutant strain responded to dietary supplementation with purified exogenous Q(10). Here we show that the Q(9) content can be determined in lipid extracts from just 200 individual worms, enabling the determination of Q content in the coq-3 mutants unable to reproduce. An extra-chromosomal array expressing wild-type C. elegans coq-3 rescued fertility of both coq-3 mutants and partially restored steady-state levels of COQ-3 polypeptide and Q(9) content, indicating that primary defect in both is limited to coq-3. The limited response of the coq-3 mutants to dietary supplementation with Q provides a powerful model to probe the effectiveness of exogenous Q supplementation as compared to restoration of de novo Q biosynthesis.

Topics: Amino Acid Sequence; Animals; Base Sequence; Caenorhabditis elegans; Caenorhabditis elegans Proteins; DNA, Helminth; Escherichia coli; Escherichia coli Proteins; Female; Fertility; Genes, Helminth; Genetic Complementation Test; Homozygote; Male; Methyltransferases; Models, Biological; Molecular Sequence Data; Mutation; Phenotype; Recombinant Proteins; Sequence Deletion; Sequence Homology, Amino Acid; Ubiquinone

Mclk1 (also known as Coq7) and Coq3 code for mitochondrial enzymes implicated in the biosynthetic pathway of ubiquinone (coenzyme Q or UQ). Mclk1(+/-) mice are long-lived but have dysfunctional mitochondria. This phenotype remains unexplained, as no changes in UQ content were observed in these mutants. By producing highly purified submitochondrial fractions, we report here that Mclk1(+/-) mice present a unique mitochondrial UQ profile that was characterized by decreased UQ levels in the inner membrane coupled with increased UQ in the outer membrane. Dietary-supplemented UQ(10) was actively incorporated in both mitochondrial membranes, and this was sufficient to reverse mutant mitochondrial phenotypes. Further, although homozygous Coq3 mutants die as embryos like Mclk1 homozygous null mice, Coq3(+/-) mice had a normal lifespan and were free of detectable defects in mitochondrial function or ubiquinone distribution. These findings indicate that MCLK1 regulates both UQ synthesis and distribution within mitochondrial membranes.

Topics: Animals; Cell Respiration; Male; Membrane Proteins; Mice; Mice, Inbred BALB C; Mice, Transgenic; Mitochondria; Mitochondrial Membranes; Mitochondrial Proteins; Mixed Function Oxygenases; Oxygen Consumption; Submitochondrial Particles; Ubiquinone

Two novel gram-negative, oxidase- and catalase-positive, rod-shaped bacterial strains, designated YCSA28(T) and YCSA39, were isolated from sediment of Daqiao saltern, Jimo, Qingdao, on the east coast of China. The two strains grew optimally at 28-30 °C, at pH 7.5 and in the presence of 7-8 % (w/v) NaCl. They were assigned to the genus Halomonas, class Gammaproteobacteria, based on 16S rRNA gene sequence analysis. The major cellular fatty acids of the two strains were C(18 : 1)ω7c (42.9 %), C(16 : 0) (23.1 %) and C(16 : 1)ω7c/ω6c (18.0 %), and Q-9 was the major ubiquinone. The G+C content of the DNA of strains YCSA28(T) and YCSA39 was 63.7 and 63.9 mol%, respectively. The predominant respiratory lipoquinone, cellular fatty acid profiles and DNA G+C content of strains YCSA28(T) and YCSA39 were consistent with those of recognized species of the genus Halomonas. Levels of DNA-DNA relatedness between strains YCSA28(T) and YCSA39, between YCSA28(T) and Halomonas ventosae Al12(T), and between YCSA39 and H. ventosae Al12(T) were 95, 45 and 50 %, respectively. Together, these data indicated that strains YCSA28(T) and YCSA39 represent a single novel species of the genus Halomonas, for which the name Halomonas daqiaonensis sp. nov. is proposed. The type strain is YCSA28(T) ( = CGMCC 1.9150(T)  = NCCB 100305(T)  = MCCC 1B00920(T)).

Topics: Bacterial Typing Techniques; Base Composition; China; Denitrification; DNA, Bacterial; Fatty Acids; Halomonas; Molecular Sequence Data; Nucleic Acid Hybridization; Phylogeny; RNA, Ribosomal, 16S; Salinity; Sequence Analysis, DNA; Sodium Chloride; Ubiquinone; Water Microbiology

Mitochondrial disorders are often associated with primary or secondary CoQ10 decrease. In clinical practice, Coenzyme Q10 (CoQ10) levels are measured to diagnose deficiencies and to direct and monitor supplemental therapy. CoQ10 is reduced by complex I or II and oxidized by complex III in the mitochondrial respiratory chain. Therefore, the ratio between the reduced (ubiquinol) and oxidized (ubiquinone) CoQ10 may provide clinically significant information in patients with mitochondrial electron transport chain (ETC) defects. Here, we exploit mutants of Caenorhabditis elegans (C. elegans) with defined defects of the ETC to demonstrate an altered redox ratio in Coenzyme Q9 (CoQ9), the native quinone in these organisms. The percentage of reduced CoQ9 is decreased in complex I (gas-1) and complex II (mev-1) deficient animals, consistent with the diminished activity of these complexes that normally reduce CoQ9. As anticipated, reduced CoQ9 is increased in the complex III deficient mutant (isp-1), since the oxidase activity of the complex is severely defective. These data provide proof of principle of our hypothesis that an altered redox status of CoQ may be present in respiratory complex deficiencies. The assessment of CoQ10 redox status in patients with mitochondrial disorders may be a simple and useful tool to uncover and monitor specific respiratory complex defects.

Topics: Animals; Antioxidants; Caenorhabditis elegans; Disease Models, Animal; Gas Chromatography-Mass Spectrometry; Humans; Mitochondria; Mitochondrial Diseases; Oxidation-Reduction; Ubiquinone

We studied ubiquinone (Q), Q homologue ratio, and steady-state levels of mCOQ transcripts in tissues from mice fed ad libitum or under calorie restriction. Maximum ubiquinone levels on a protein basis were found in kidney and heart, followed by liver, brain, and skeletal muscle. Liver and skeletal muscle showed the highest Q(9)/Q(10) ratios with significant interindividual variability. Heart, kidney, and particularly brain exhibited lower Q(9)/Q(10) ratios and interindividual variability. In skeletal muscle and heart, the most abundant mCOQ transcript was mCOQ7, followed by mCOQ8, mCOQ2, mPDSS2, mPDSS1, and mCOQ3. In nonmuscular tissues (liver, kidney, and brain) the most abundant mCOQ transcript was mCOQ2, followed by mCOQ7, mCOQ8, mPDSS1, mPDSS2, and mCOQ3. Calorie restriction increased both ubiquinone homologues and mPDSS2 mRNA in skeletal muscle, but mCOQ7 was decreased. In contrast, Q(9) and most mCOQ transcripts were decreased in heart. Calorie restriction also modified the Q(9)/Q(10) ratio, which was increased in kidney and decreased in heart without alterations in mPDSS1 or mPDSS2 transcripts. We demonstrate for the first time that unique patterns of mCOQ transcripts exist in muscular and nonmuscular tissues and that Q and COQ genes are targets of calorie restriction in a tissue-specific way.

Topics: Animals; Brain; Caloric Restriction; Free Radicals; Kidney; Liver; Mice; Muscle, Skeletal; Myocardium; Organ Specificity; RNA, Messenger; Ubiquinone

Virgin argan oil possesses high antioxidant capacity (AC), which may be partially explained by its high content in antioxidant molecules such as polyphenols and tocopherols. However, the content in other antioxidant molecules, for example, coenzyme Q10 (CoQ(10)), coenzyme Q9 (CoQ(9)), and melatonin (Mel), which have been identified in other edible vegetable oils, have not been evaluated in virgin argan oil. Consequently, it was decided to evaluate the contents of CoQ(10), CoQ(9), and Mel in virgin argan oils and compare the results to those obtained in extra virgin olive oils and some varieties of seed oils. By the use of sensitive HPLC-EC/F methods, the results showed that virgin argan oil is a rich source of CoQ(10) and Mel, but no CoQ(9) was detected. Extra virgin olive oil showed higher levels of CoQ(10) and lower levels of Mel than virgin argan oil. Between the seed oil samples, only virgin soybean oil showed higher CoQ(10) and Mel levels than virgin argan oil. The results may be relevant for the contribution of CoQ(10) and Mel to the biological activities of virgin argan oil.

Topics: Chromatography, High Pressure Liquid; Melatonin; Plant Extracts; Plant Oils; Sapotaceae; Ubiquinone

It has been hypothesized that oxidative stress plays a key role in aging. In order to elucidate the role of the antioxidant network - including alpha-tocopherol (alphaT) and alphaT transfer protein - in aging in vivo, alpha-tocopherol transfer protein knockout (alphaTTP(-/-)) mice were fed a vitamin-E-depleted diet, and wild-type (WT) mice were fed a diet containing 0.002 wt.% alphaT from the age of 3 months to 1 1/2 years. The lipid oxidation markers total hydroxyoctadecadienoic acid (tHODE) and 8-iso-prostaglandin F(2)alpha, and antioxidant levels in the blood, liver and brain were measured at 3, 6, 12 and 18 months. tHODE levels in the plasma of alphaTTP(-/-) mice were elevated at 6 months compared to 3 months, and were significantly higher those in WT mice, although they decreased thereafter. On the other hand, tHODE levels in the liver and brain were constantly higher in alphaTTP(-/-) mice than in WT mice. Motor activities decreased with aging in both mouse types; however, those in the alphaTTP(-/-) mice were lower than those in the WT mice. It is intriguing to note that motor activities were significantly correlated with the stereoisomer ratio (Z,E/E,E) of HODE, which is a measure of antioxidant capacity in vivo, in the plasma, in the liver and even in the brain, but not with other factors such as antioxidant levels. In summary, using the biomarker tHODE and its stereoisomer ratio, we demonstrated that alphaT depletion was associated with a decrease in motor function, and that this may be primarily attributable to a decrease in the total antioxidant capacity in vivo.

Topics: Aging; alpha-Tocopherol; Animals; Ascorbic Acid; Biomarkers; Brain Chemistry; Carrier Proteins; Dinoprost; Fatty Acids, Unsaturated; Female; Lipid Peroxidation; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Motor Activity; Oxidative Stress; Specific Pathogen-Free Organisms; Stereoisomerism; Thiobarbituric Acid Reactive Substances; Ubiquinone; Vitamin E Deficiency

Oxidative stress has been associated with many pathological disorders such as atherosclerosis, diabetes and cancer. Supplementation with exogenous antioxidants, including phenolic compounds from plant sources, may help to restore the pro-oxidative/antioxidative balance. To take into account effects of absorption, metabolisation, plasma protein binding, distribution, and elimination, antioxidative research should not be limited to in vitro assays but be extended to in vivo models.. In the present work a quantified 50% EtOH root extract of Pueraria lobata (Willd.) Ohwi (Fabaceae) was selected to determine its in vivo antioxidative activity in a diabetic rat model, where diabetes and the accompanying oxidative stress were induced by intraperitoneal administration of streptozotocin. This root extract was found to contain 10.42+/-0.15% puerarin as the main constituent and smaller amounts of the related isoflavonoids 3'-hydroxypuerarin, 3'-methoxypuerarin, 6''-xylosylpuerarin, daidzin, genistin, daidzein and genistein, as determined by a validated HPLC method. This extract was administered orally at a daily dose of 500 mg/kg root extract, corresponding to 50mg/kg puerarin, during 3 weeks. In addition the effect on the plasma concentration of some fat-soluble antioxidants (co-enzyme Q(9), alpha- and gamma-tocopherol) was evaluated.. The level of malondialdehyde (MDA) in plasma, used as a marker of oxidative damage to lipids, was reduced to the same level as in healthy control animals, and as in the positive control group treated daily with 50mg/kg alpha-tocopherol acetate. No obvious signs of toxicity were observed by administration of 10x the treatment dose.

Topics: alpha-Tocopherol; Animals; Antioxidants; Blood Glucose; Body Weight; Diabetes Mellitus, Experimental; Drugs, Chinese Herbal; gamma-Tocopherol; Isoflavones; Lipid Peroxidation; Male; Malondialdehyde; Oxidative Stress; Plant Extracts; Plant Roots; Pueraria; Rats; Rats, Wistar; Ubiquinone

A Gram-negative, heterotrophic, aerobic, non-endospore-forming, peritrichously flagellated and motile bacterial strain, designated BH1(T), was isolated from samples of rusticles, which are formed in part by a consortium of micro-organisms, collected from the RMS Titanic wreck site. The strain grew optimally at 30-37°C, pH 7.0-7.5 and in the presence of 2-8 % (w/v) NaCl. We carried out a polyphasic taxonomic study in order to characterize the strain in detail. Phylogenetic analyses based on 16S rRNA gene sequence comparison indicated that strain BH1(T) clustered within the branch consisting of species of Halomonas. The most closely related type strains were Halomonas neptunia (98.6 % 16S rRNA sequence similarity), Halomonas variabilis (98.4 %), Halomonas boliviensis (98.3 %) and Halomonas sulfidaeris (97.5 %). Other closely related species were Halomonas alkaliphila (96.5 % sequence similarity), Halomonas hydrothermalis (96.3 %), Halomonas gomseomensis (96.3 %), Halomonas venusta (96.3 %) and Halomonas meridiana (96.2 %). The major fatty acids of strain BH1(T) were C(18 : 1)ω7c (36.3 %), C(16 : 0) (18.4 %) and C(19 : 0) cyclo ω8c (17.9 %). The DNA G+C content was 60.0 mol% (T(m)). Ubiquinone 9 (Q-9) was the major lipoquinone. The phenotypic features, fatty acid profile and DNA G+C content further supported the placement of strain BH1(T) in the genus Halomonas. DNA-DNA hybridization values between strain BH1(T) and H. neptunia CECT 5815(T), H. variabilis DSM 3051(T), H. boliviensis DSM 15516(T) and H. sulfidaeris CECT 5817(T) were 19, 17, 30 and 29 %, respectively, supporting the differential taxonomic status of BH1(T). On the basis of the phenotypic, chemotaxonomic and phylogenetic data, strain BH1(T) is considered to represent a novel species, for which the name Halomonas titanicae sp. nov. is proposed. The type strain is BH1(T) (=ATCC BAA-1257(T) =CECT 7585(T) =JCM 16411(T) =LMG 25388(T)).

Topics: Atlantic Ocean; Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Halomonas; Molecular Sequence Data; Nucleic Acid Hybridization; Phylogeny; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Ships; Ubiquinone

A novel Gram-negative, aerobic, motile, short rod-shaped bacterium, designated MS-3(T), was isolated from a crude oil-contaminated seashore in Taean, Korea. Strain MS-3(T) grew at 4-30 °C, at pH 6.0-9.5 and with 0-5 % NaCl and was oxidase- and catalase-positive. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain MS-3(T) was most similar to Pseudomonas marincola KMM 3042(T) (97.9 % 16S rRNA gene sequence similarity), P. cuatrocienegasensis 1N(T) (97.8 %), P. borbori R-20821(T) (97.3 %) and P. lundensis ATCC 49968(T) (97.1 %). Relatively low levels of DNA-DNA relatedness were found between strain MS-3(T) and P. cuatrocienegasensis LMG 24676(T) (57.2 %), P. borbori LMG 23199(T) (39.7 %), P. marincola KMM 3042(T) (32.2 %) and P. lundensis KACC 10832(T) (32.1 %), which support the classification of strain MS-3(T) within a novel species of the genus Pseudomonas. The G+C content of the genomic DNA of strain MS-3(T) was 57.6 mol% and the major isoprenoid quinone was Q-9. Strain MS-3(T) contained summed feature 3 (iso-C(15 : 0) 2-OH and/or C(16 : 1)ω7c; 38.0 %), C(16 : 0) (24.4 %), C(18 : 1)ω7c (12.8 %), C(12 : 0) (9.6 %) and C(10 : 0) 3-OH (4.9 %) as the major cellular fatty acids. On the basis of the phenotypic, genotypic and phylogenetic data, strain MS-3(T) represents a novel species of the genus Pseudomonas, for which the name Pseudomonas taeanensis sp. nov. is proposed. The type strain is MS-3(T) (=KCTC 22612(T) =KACC 14032(T) =JCM 16046(T) =NBRL 105641(T)).

Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; DNA, Ribosomal; Fatty Acids; Molecular Sequence Data; Nucleic Acid Hybridization; Petroleum; Petroleum Pollution; Phylogeny; Pseudomonas; Republic of Korea; RNA, Ribosomal, 16S; Seawater; Sequence Analysis, DNA; Ubiquinone

Diets high in saturated fat and fructose have been implicated in the development of obesity and nonalcoholic steatohepatitis (NASH) in humans. We hypothesized that mice exposed to a similar diet would develop NASH with fibrosis associated with increased hepatic oxidative stress that would be further reflected by increased plasma levels of the respiratory chain component, oxidized coenzyme Q9 ((ox)CoQ9). Adult male C57Bl/6 mice were randomly assigned to chow, high-fat (HF), or high-fat high-carbohydrate (HFHC) diets for 16 weeks. The chow and HF mice had free access to pure water, whereas the HFHC group received water with 55% fructose and 45% sucrose (wt/vol). The HFHC and HF groups had increased body weight, body fat mass, fasting glucose, and were insulin-resistant compared with chow mice. HF and HFHC consumed similar calories. Hepatic triglyceride content, plasma alanine aminotransferase, and liver weight were significantly increased in HF and HFHC mice compared with chow mice. Plasma cholesterol (P < 0.001), histological hepatic fibrosis, liver hydroxyproline content (P = 0.006), collagen 1 messenger RNA (P = 0.003), CD11b-F4/80+Gr1+ monocytes (P < 0.0001), transforming growth factor beta1 mRNA (P = 0.04), and alpha-smooth muscle actin messenger RNA (P = 0.001) levels were significantly increased in HFHC mice. Hepatic oxidative stress, as indicated by liver superoxide expression (P = 0.002), 4-hydroxynonenal, and plasma (ox)CoQ9 (P < 0.001) levels, was highest in HFHC mice.. These findings demonstrate that nongenetically modified mice maintained on an HFHC diet in addition to developing obesity have increased hepatic ROS and a NASH-like phenotype with significant fibrosis. Plasma (ox)CoQ9 correlated with fibrosis progression. The mechanism of fibrosis may involve fructose inducing increased ROS associated with CD11b+F4/80+Gr1+ hepatic macrophage aggregation, resulting in transforming growth factor beta1-signaled collagen deposition and histologically visible hepatic fibrosis.

Topics: Animals; Body Composition; Collagen; Dietary Carbohydrates; Dietary Fats; Disease Models, Animal; Fatty Liver; Fructose; Insulin Resistance; Liver; Liver Cirrhosis; Male; Mice; Mice, Inbred C57BL; Obesity; Reactive Oxygen Species; Trans Fatty Acids; Transforming Growth Factor beta; Ubiquinone

Rheumatoid arthritis is a common severe joint disease that affects all age groups, it is thus of great importance to develop new strategies for its treatment. The aim of the present study was to examine the combined effect of coenzyme Q₁₀ (CoQ₁₀) and methotrexate (MTX) on the progression of adjuvant-induced arthritis in rats. Adjuvant arthritis (AA) was induced by a single intradermal injection of heat-inactivated Mycobacterium butyricum in incomplete Freund's adjuvant. The experiments included healthy animals, arthritic animals not treated, arthritic animals treated with CoQ₁₀, with methotrexate, and with a combination of CoQ₁₀ and methotrexate. The two latter groups received a daily oral dose of 20 mg/kg b.w. of CoQ₁₀, either alone or with methotrexate in an oral dose of 0.3 mg/kg b.w. twice a week. We found that CoQ₁₀ potentiated both the antiarthritic (decrease of hind paw volume) and the antioxidant effect of methotrexate on the level of oxidation of proteins (suppression of protein carbonyl level in plasma) as well as lipoperoxidation (suppression of levels of HNE-adducts and MDA-adducts to plasma proteins). The same effect was observed for plasmatic levels of CoQ₉ and IL-1α, and partially also for γ-glutamyltransferase activity assessed in joints and spleen. Moreover, the combination therapy improved the functionality of peripheral blood neutrophils in AA, with a balancing effect on the immunosuppression caused by MTX monotherapy. In summary, combined administration of CoQ₁₀ and methotrexate suppressed arthritic progression in rats more effectively than did MTX alone. This finding may help improve treatment of rheumatoid arthritis.

Topics: Animals; Arthritis, Experimental; Drug Therapy, Combination; Enzyme-Linked Immunosorbent Assay; Inflammation; Interleukin-1alpha; Lipid Peroxidation; Male; Methotrexate; Oxidative Stress; Rats; Ubiquinone

Coenzyme Q (Q) regulates aging in Caenorhabditis elegans, and its deficiency leads to a variety of pathologies in humans. We used a coq-8 deleted strain to study the role of Q in C. elegans development and how it influences life span. Endogenous Q(9) content of coq-8(ok840) knockouts was demonstrated to be about 7% of that found in the wild-type, indicating the basal biosynthesis rate is reduced in this strain. Knockouts abnormally developed both gonads and hypodermis, showed reduced fertility and shortened life span, and this was partially recovered by ingestion of exogenous Q. Knockouts produced embryos that showed arrested development at the time of initial expression of coq-8 in embryo. Uridine, whose biosynthesis depends on mitochondrial Q, improved both egg production and progeny under Q-rich dietary conditions. COQ-8 is a candidate protein for post-translational regulation of Q biosynthesis rate and its expression correlates with Q content during the life cycle in C. elegans. We show for the first time that a critical level of Q is necessary to support embryo development and fertility in C. elegans. These results suggest that extra-mitochondrial function of Q is a key factor linking development and bioenergetics in C. elegans.

Topics: Aging; Animals; Caenorhabditis elegans; Caenorhabditis elegans Proteins; Fertility; Gene Expression Regulation, Developmental; Gene Expression Regulation, Enzymologic; Gene Knockout Techniques; Genotype; Gonads; Larva; Longevity; Phenotype; Ubiquinone; Uridine

Coenzyme Q(10) is a mobile lipophilic electron carrier located in the inner mitochondrial membrane. Defects of coenzyme Q(10) biosynthesis represent one of the few treatable mitochondrial diseases. We genotyped a patient with primary coenzyme Q(10) deficiency who presented with neonatal lactic acidosis and later developed multisytem disease including intractable seizures, global developmental delay, hypertrophic cardiomyopathy, and renal tubular dysfunction. Cultured skin fibroblasts from the patient had a coenzyme Q(10) biosynthetic rate of 11% of normal controls and accumulated an abnormal metabolite that we believe to be a biosynthetic intermediate. In view of the rarity of coenzyme Q(10) deficiency, we hypothesized that the disease-causing gene might lie in a region of ancestral homozygosity by descent. Data from an Illumina HumanHap550 array were analyzed with BeadStudio software. Sixteen regions of homozygosity >1.5 Mb were identified in the affected infant. Two of these regions included the loci of two of 16 candidate genes implicated in human coenzyme Q(10) biosynthesis. Sequence analysis demonstrated a homozygous stop mutation affecting a highly conserved residue of COQ9, leading to the truncation of 75 amino acids. Site-directed mutagenesis targeting the equivalent residue in the yeast Saccharomyces cerevisiae abolished respiratory growth.

Topics: Amino Acid Sequence; Cells, Cultured; Codon, Nonsense; Fibroblasts; Genetic Predisposition to Disease; Homozygote; Humans; Infant; Infant, Newborn; Mitochondrial Diseases; Models, Molecular; Molecular Sequence Data; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins; Skin; Ubiquinone

Ubiquinone (UQ, Coenzyme Q, CoQ) transfers electrons from complexes I and II to complex III in the mitochondrial electron transport chain. Depending on the degree of reduction, UQ can act as either a pro- or an antioxidant. Mutations disrupting ubiquinone synthesis increase lifespan in both the nematode (clk-1) and the mouse (mclk-1). The mutated nematodes survive using exogenous ubiquinone from bacteria, which has a shorter isoprenyl tail length (UQ(8)) than the endogenous nematode ubiquinone (UQ(9)). The mechanism underlying clk-1s increased longevity is not clear. Here we directly measure the effect of different exogenous ubiquinones on clk-1 lifespan and mitochondrial function. We fed clk-1 engineered bacteria that produced UQ(6), UQ(7), UQ(8), UQ(9) or UQ(10), and measured clk-1s lifespan, mitochondrial respiration, ROS production, and accumulated ROS damage to mitochondrial protein. Regardless of dietary UQ, clk-1 animals have increased lifespan, decreased mitochondrial respiration, and decreased ROS damage to mitochondrial protein than N2. However, clk-1 mitochondria did not produce less ROS than N2. The simplest explanation of our results is that clk-1 mitochondria scavenge ROS more effectively than wildtype due to the presence of DMQ(9). Moreover, when compared to other dietary quinones, UQ(10) further decreased mitochondrial oxidative damage and extended adult lifespan in clk-1.

Topics: Animals; Caenorhabditis elegans; Caenorhabditis elegans Proteins; Cell Respiration; Hydrogen Peroxide; Longevity; Mitochondria; Mitochondrial Proteins; Mutation; Oxidative Stress; Reactive Oxygen Species; Time Factors; Ubiquinone

We recently reported insulin resistance in adult offspring of obese C57BL/6J mice. We have now evaluated whether parameters of skeletal muscle structure and function may play a role in insulin resistance in this model of developmental programming. Obesity was induced in female mice by feeding a highly palatable sugar and fat-rich diet for 6 wk prior to pregnancy, and during pregnancy and lactation. Offspring of obese dams were weaned onto standard laboratory chow. At 3 mo of age, skeletal muscle insulin signaling protein expression, mitochondrial electron transport chain activity (ETC), muscle fiber type, fiber density, and fiber cross-sectional area were compared with that of offspring of control dams weaned onto the chow diet. Female offspring of obese dams demonstrated decreased skeletal muscle expression of p110beta, the catalytic subunit of PI3K (P < 0.01), as well as reduced Akt phosphorylation at Serine residue 473 compared with control offspring. Male offspring of obese dams demonstrated increased skeletal muscle Akt2 and PKCzeta expression (P < 0.01; P < 0.001, respectively). A decrease in mitochondrial-linked complex II-III was observed in male offspring of obese dams (P < 0.01), which was unrelated to CoQ deficiency. This was not observed in females. There were no differences in muscle fiber density between offspring of obese dams and control offspring in either sex. Sex-related alterations in key insulin-signaling proteins and in mitochondrial ETC may contribute to a state of insulin resistance in offspring of obese mice.

Topics: Animal Nutritional Physiological Phenomena; Animals; Body Weight; Class I Phosphatidylinositol 3-Kinases; Disease Models, Animal; Electron Transport Complex II; Electron Transport Complex III; Female; Glucose Transporter Type 4; Insulin; Insulin Receptor Substrate Proteins; Insulin Resistance; Male; Maternal Nutritional Physiological Phenomena; Mice; Mice, Inbred C57BL; Mitochondria, Muscle; Muscle Fibers, Skeletal; Obesity; Phosphatidylinositol 3-Kinases; Phosphorylation; Pregnancy; Prenatal Exposure Delayed Effects; Protein Kinase C; Proto-Oncogene Proteins c-akt; Quadriceps Muscle; Receptor, Insulin; Sex Factors; Signal Transduction; Ubiquinone

The overproduction of very-low-density lipoprotein (VLDL) is a characteristic feature of nonalcoholic fatty liver disease (NAFLD). The aim of this study was to use a high-fat diet-induced model of NAFLD in rats to investigate 1) the influence of the disease on hepatic VLDL processing in the endoplasmic reticulum and 2) the potential modulatory effects of dietary coenzyme Q (CoQ). Rats were fed a standard low-fat diet (control) or a diet containing 35% fat (57% metabolizable energy). After 10 wk, high-fat diet-fed animals were divided into three groups: the first group was given CoQ9 (30 mg*kg body wt(-1)*day(-1) in 0.3 ml olive oil), the second group was given olive oil (0.3 ml/day) only, and the third group received no supplements. Feeding (3 high-fat diets and the control diet) was then continued for 8 wk. In all high-fat diet-fed groups, the content of triacylglycerol (TG) and cholesterol in plasma VLDL, the liver, and liver microsomes was increased, hepatic levels of apolipoprotein B48 were raised, and the activities of microsomal TG transfer protein and acyl CoA:cholesterol acyltransferase were reduced. These findings provide new evidence indicating that VLDL assembly and the inherent TG transfer to the endoplasmic reticulum are altered in NAFLD and suggest a possible explanation for both the overproduction of VLDL associated with the condition and the disease etiology itself. Dietary CoQ caused significant increases in apolipoprotein B mRNA and microsomal TG levels and altered the phospholipid content of microsomal membranes. These changes, however, may not be beneficial as they may lead to the secretion of larger, more atherogenic VLDL.

Topics: Animals; Antioxidants; Apolipoproteins B; Diet; Fatty Liver; Lipid Metabolism; Lipoproteins, VLDL; Liver; Male; Microsomes, Liver; Rats; Rats, Wistar; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Ubiquinone

A chloroform extract of the edible mushroom Pleurotus eryngii showed an inhibitory effect on mammalian DNA topoisomerase I. The topoisomerase I inhibitory compound was purified and identified as ubiquinone-9. Ubiquinone-9 was shown to inhibit the activity of topoisomerase I with IC(50) of about 50 microM. Concentration of 110 microM ubiquinone-9 caused 50% growth inhibition of human leukaemia U937 cells, but not that of normal fibroblast NIH3T3 and 3Y1 cells. Ubiquinone-9-induced cell death was characterised with the cleavage of poly (ADP-ribose) polymerase and pro-caspase 3. Furthermore, ubiquinone-9 induced the fragmentation of DNA into an apoptotic DNA ladder, indicating that the inhibitor triggered apoptosis. The induction of apoptosis by ubiquinone-9 was also confirmed using flow cytometry analysis. Taken together, these results suggest that ubiquinone-9 may function by inhibiting oncogenic disease, at least in part, through the inhibition of topoisomerase I activity.

Topics: Antineoplastic Agents; Apoptosis; Blotting, Western; DNA Fragmentation; DNA Topoisomerases, Type I; Enzyme Activation; Flow Cytometry; Humans; Molecular Structure; Pleurotus; U937 Cells; Ubiquinone

Coenzyme Q(10) (CoQ(10)) is widely consumed as a dietary supplement to enhance bioenergetic capacity and to ameliorate the debilitative effects of the aging process or certain pathological conditions. Our main purpose in this study was to determine whether CoQ(10) intake does indeed attenuate the age-associated losses in motor, sensory, and cognitive functions or decrease the rate of mortality in mice. Mice were fed a control nonpurified diet or that diet containing 0.68 mg/g (low dosage) or 2.6 mg/g (high dosage) CoQ(10), starting at 4 mo of age, and were tested for sensory, motor, and cognitive function at 7, 15, and 25 mo of age. Amounts of the ubiquinols CoQ(9)H(2) and CoQ(10)H(2) measured in a parallel study were augmented in the cerebral cortex but not in any other region of the brain. Intake of the low-CoQ(10) diet did not affect age-associated decrements in muscle strength, balance, coordinated running, or learning/memory, whereas intake at the higher amount increased spontaneous activity, worsened the age-related losses in acuity to auditory and shock stimuli, and impaired the spatial learning/memory of old mice. The CoQ(10) diets did not affect survivorship of mice through 25 mo of age. Our results suggest that prolonged intake of CoQ(10) in low amounts has no discernable impact on cognitive and motor functions whereas intake at higher amounts exacerbates cognitive and sensory impairments encountered in old mice. These findings do not support the notion that CoQ(10) is a fitness-enhancing or an "antiaging" substance under normal physiological conditions.

Topics: Aging; Animals; Body Weight; Brain; Brain Chemistry; Cognition; Dietary Supplements; Drug Administration Schedule; Eating; Male; Mice; Mice, Inbred C57BL; Ubiquinone

Coenzyme Q10 (CoQ10) has been extensively studied as adjunctive therapy for ischemic heart disease, and its cardioprotective ability is well-established. The mitochondrial respiratory chain contains several coenzymes, including CoQ1, CoQ2, CoQ4, CoQ6, CoQ7, CoQ8, CoQ9, and CoQ10. It is not known whether other CoQs, especially CoQ9, is equally cardioprotective as CoQ10. The present study was designed to determine if CoQ 9 could protect guinea pig hearts from ischemia reperfusion injury. Guinea pigs were randomly divided into three groups: groups I and II were fed CoQ 9 and CoQ10, respectively, for 30 days while group III served as control. After 30 days, the guinea pigs were sacrificed and isolated hearts were perfused via working mode were subjected to 30 min ischemia followed by 2 h of reperfusion. Cardioprotection was assessed by evaluating left ventricular function, ventricular arrhythmias, myocardial infarct size, and cardiomyocyte apoptosis. Samples of hearts were examined for the presence of CoQ9 and CoQ10. The results demonstrated that both CoQ9 and CoQ10 were equally cardioprotective, as evidenced by their abilities to improve left ventricular performance and to reduce myocardial infarct size and cardiomyocyte apoptosis. High performance liquid chromatographic (HPLC) analysis revealed that a substantial portion of CoQ9 had been converted into CoQ10. The results indicate that CoQ9 by itself, or after being converted into CoQ10, reduced myocardial ischemia/reperfusion-induced injury.

Topics: Animals; Apoptosis; Arrhythmias, Cardiac; Biotransformation; Cardiotonic Agents; Drug Evaluation, Preclinical; Guinea Pigs; Humans; Male; Mass Spectrometry; Myocardial Reperfusion Injury; Random Allocation; Ubiquinone; Ventricular Function, Left

A new sensitive and selective method has been developed for the quantification of the total coenzyme Q9 (CoQ9) and coenzyme Q10 (CoQ10) concentration in vegetable oil samples. The coenzyme Q fraction is isolated by solid-phase extraction (SPE) on amino phase eluting with a mixture of heptane:ethyl ether. The organic solvent is evaporated under nitrogen, and the residue is dissolved in a mixture of acetonitrile:tetrahydrofuran and finally is analyzed by reverse-phase high-performance liquid chromatography with a mass detector. The sensitivity of the method is based on the high efficient formation of the radical anions [M (-.)] of CoQ9 and CoQ10 by negative atmospheric pressure ionization. Interferences are minimized by using mass detection of the [M (-.)] ions ( m/ z = 797.5 for CoQ9 and m/ z = 862.5 for CoQ10) in selective reaction monitoring mode ( m/ z = 797.5 --> m/ z = 779.5 and m/ z = 862.5 --> m/ z = 847.5) using a triple-quadrupole mass spectrometer. The method was successfully applied to sunflower, soybean, and rapeseed oils, with a limit of quantification of 0.025 mg/kg for both compounds.

Topics: Chromatography, High Pressure Liquid; Fatty Acids, Monounsaturated; Plant Oils; Rapeseed Oil; Sensitivity and Specificity; Solvents; Soybean Oil; Sunflower Oil; Ubiquinone

Homozygous mice carrying kd (kidney disease) mutations in the gene encoding prenyl diphosphate synthase subunit 2 (Pdss2kd/kd) develop interstitial nephritis and eventually die from end-stage renal disease. The PDSS2 polypeptide in concert with PDSS1 synthesizes the polyisoprenyl tail of coenzyme Q (Q or ubiquinone), a lipid quinone required for mitochondrial respiratory electron transport. We have shown that a deficiency in Q content is evident in Pdss2kd/kd mouse kidney lipid extracts by 40 days of age and thus precedes the onset of proteinuria and kidney disease by several weeks. The presence of the kd (V117M) mutation in PDSS2 does not prevent its association with PDSS1. However, heterologous expression of the kd mutant form of PDSS2 together with PDSS1 in Escherichia coli recapitulates the Q deficiency observed in the Pdss2kd/kd mouse. Dietary supplementation with Q10 provides a dramatic rescue of both proteinuria and interstitial nephritis in the Pdss2kd/kd mutant mice. The results presented suggest that Q may be acting as a potent lipid-soluble antioxidant, rather than by boosting kidney mitochondrial respiration. Such Q10 supplementation may have profound and beneficial effects in treatment of certain forms of focal segmental glomerulosclerosis that mirror the renal disease of the Pdss2kd/kd mouse.

Topics: Albuminuria; Alkyl and Aryl Transferases; Animals; Dietary Supplements; Female; Gene Expression; Glomerulosclerosis, Focal Segmental; Kidney; Liver; Male; Mice; Mice, Inbred Strains; Mice, Mutant Strains; Mice, Transgenic; Mitochondria; Mutation; Nephritis; Protein Binding; Protein Subunits; Transfection; Ubiquinone

This chapter describes the use of reversed-phase HPLC with multichannel coulometric electrochemical detection for the routine, sensitive, and simultaneous measurement of oxidized and reduced CoQ10 and CoQ9 in human plasma and serum. Analytes are first resolved chromatographically prior to electrochemical detection using three serially placed flow-through coulometric sensors set for oxidation-reduction-re-oxidation. Such electrochemical manipulation of analytes not only improves selectivity and specificity (decreasing the likelihood of co-elution), but also leads to improved sensitivity and decreased noise. The method is completed in ,18 min, shows excellent linearity, good intra-day (% RSD = 1.2-2.3) and inter-day (% RSD 2.2-3.9) precision, and has a limit of detection to low pg levels (on column). This approach was used to measure oxidized and reduced CoQ10 and CoQ9 in 30 human plasma samples, and oxidized and reduced CoQ10 in 10 human serum samples (NIST Micronutrients Measurement Quality Assurance Program for fat-soluble vitamins).

Topics: Calibration; Chromatography, High Pressure Liquid; Humans; Oxidation-Reduction; Quality Control; Reference Standards; Time Factors; Ubiquinone

Effect of CoQ compounds (Qs) on reactive oxygen (ROS) generation by mitochondrial complex I was studied using rat liver mitochondria and chemiluminescence probe L012. Kinetic analysis revealed that short chain Qs, such as Q2 and idebenone enhanced ROS generation by mitochondrial NADH oxidase system by a succinate-inhibitable mechanism. Lipid peroxidation in mitochondrial membranes induced by NADH and iron was inhibited by short chain Qs. The inhibitory activity was enhanced by co-oxidation of succinate as determined by chemiluminescence method and by electron spin resonance spectroscopy. These results suggested that the reduced form of short chain Qs inhibited mitochondrial ROS generation and lipid peroxidation.

Topics: Animals; Male; Metabolic Networks and Pathways; Mitochondria, Liver; Oxidoreductases; Rats; Rats, Wistar; Reactive Oxygen Species; Ubiquinone

A UPLC-MS method for determining Coenzyme Q(10) (CoQ(10)) levels in rat serum was developed. CoQ(10) was quantitatively extracted into 2-propanol using a fast extraction procedure. The separation of CoQ(10) was performed on a Waters Acquity UPLCtrade mark BEH C(18) column (1.7 microm, 1.0 mm x 50 mm) with the mobile phase containing acetonitrile, 2-propanol, and formic acid (90:10:0.1) over 5 min. The sensitivity of this method allows for the quantitation of 50 ng/mL CoQ(10) in serum (S/N=10). The linearity of this method was found to be from 50 to 20,000 ng/mL. The precision was less than 10% (intra- and inter-day), and the average extraction recovery was between 90 and 105%. This procedure provides a precise, sensitive and direct assay method for the determination of CoQ(10) in rat serum after oral administration. This method could be applied to further pharmacokinetic studies of CoQ(10).

Topics: 2-Propanol; Acetates; Acetic Acid; Acetonitriles; Administration, Oral; Ammonium Hydroxide; Animals; Area Under Curve; Chromatography, Liquid; Coenzymes; Formates; Hydroxides; Linear Models; Mass Spectrometry; Metabolic Clearance Rate; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Solvents; Ubiquinone

Statins have been shown to be cardioprotective; however, their interaction with endogenous cardioprotection by ischemic preconditioning and postconditioning is not known. In the present study, we examined if acute and chronic administration of the 3-hydroxy-3-methylglutaryl CoA reductase inhibitor lovastatin affected the infarct size-limiting effect of ischemic preconditioning and postconditioning in rat hearts. Wistar rats were randomly assigned to the following three groups: 1) vehicle (1% methylcellulose per os for 12 days), 2) chronic lovastatin (15 mg.kg(-1).day(-1) per os for 12 days), and 3) acute lovastatin (1% methylcellulose per os for 12 days and 50 micromol/l lovastatin in the perfusate). Hearts isolated from the three groups were either subjected to a nonconditioning (aerobic perfusion followed by 30-min coronary occlusion and 120-min reperfusion, i.e., test ischemia-reperfusion), preconditioning (three intermittent periods of 5-min ischemia-reperfusion cycles before test ischemia-reperfusion), or postconditioning (six cycles of 10-s ischemia-reperfusion after test ischemia) perfusion protocol. Preconditioning and postconditioning significantly decreased infarct size in vehicle-treated hearts. However, preconditioning failed to decrease infarct size in acute lovastatin-treated hearts, but the effect of postconditioning remained unchanged. Chronic lovastatin treatment abolished postconditioning but not preconditioning; however, it decreased infarct size in the nonconditioned group. Myocardial levels of coenzyme Q9 were decreased in both acute and chronic lovastatin-treated rats. Western blot analysis revealed that both acute and chronic lovastatin treatment attenuated the phoshorylation of Akt; however, acute but not chronic lovastatin treatment increased the phosphorylation of p42 MAPK/ERK. We conclude that, although lovastatin may lead to cardioprotection, it interferes with the mechanisms of cardiac adaptation to ischemic stress.

Topics: Animals; Blotting, Western; Disease Models, Animal; Down-Regulation; Drug Administration Schedule; Enzyme Activation; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Ischemic Preconditioning, Myocardial; Lovastatin; Male; Mitogen-Activated Protein Kinase 1; Myocardial Infarction; Myocardial Reperfusion Injury; Myocardium; Phosphorylation; Proto-Oncogene Proteins c-akt; Rats; Rats, Wistar; Ubiquinone

Contrary to clinical trials, experimental studies revealed that diabetes mellitus (DM) may initiate, besides increased myocardial vulnerability to ischemia-reperfusion injury (I/R) and pro/antioxidant dysbalance, development of adaptation leading to an enhanced tolerance to I/R. The aims were to characterize 1) susceptibility to ischemia-induced ventricular arrhythmias in the diabetic rat heart 2) its response to antioxidant N-acetylcysteine (NAC) and a NOS inhibitor L-NAME, and 3) the effect of DM on endogenous antioxidant systems. Seven days after streptozotocin injection (65 mg/kg, i.p.), Langendorff-perfused control (C) and DM hearts were subjected to 30-min occlusion of the LAD coronary artery with or without prior 15-min treatment with L-NAME (100 microM) or NAC (4 mM). Total number of ventricular premature beats (VPB), as well the total duration of ventricular tachycardia (VT) were reduced in the DM group (from 533+/-58 and 37.9+/-10.2 s to 224.3+/-52.6 and 19+/-13.5 s; P<0.05). In contrast to the antiarrhythmic effects of L-NAME and NAC in controls group (VPB 290+/-56 and 74+/-36, respectively; P<0.01 vs. control hearts), application of both drugs in the diabetics did not modify arrhythmogenesis (L-NAME: VPB 345+/-136, VT 25+/-13 s; NAC: VPB 207+/-50, VT 12+/-3.9 s; P>0.05 vs non-treated diabetic hearts). Diabetic state was associated with significantly elevated levels of CoQ10 and CoQ9 (19.6+/-0.8 and 217.3+/-9.5 vs. 17.4+/- 0.5 and 185.0+/-5.0 nmol/g, respectively, in controls; P<0.05), as well as alpha-tocopherol (38.6+/-0.7 vs. 31.5+/-2.1 nmol/g in controls; P<0.01) in the myocardial tissue. It is concluded that early period of DM is associated with enhanced resistance to ischemia-induced arrhythmias. Diabetes mellitus might induce adaptive processes in the myocardium leading to lower susceptibility to antioxidant and L-NAME treatment.

Topics: Acetylcysteine; Adaptation, Physiological; alpha-Tocopherol; Animals; Antioxidants; Diabetes Mellitus, Experimental; Enzyme Inhibitors; Male; Myocardial Ischemia; Myocardium; NG-Nitroarginine Methyl Ester; Nitric Oxide Synthase; Rats; Rats, Wistar; Tachycardia, Ventricular; Ubiquinone; Up-Regulation; Ventricular Function, Left

The objective of this study was to characterize the lipidome and electron transport chain activities in purified non-synaptic (NS) and synaptic (Syn) mitochondria from C57BL/6J mouse cerebral cortex. Contamination from subcellular membranes, especially myelin, has hindered past attempts to accurately characterize the lipid composition of brain mitochondria. An improved Ficoll and sucrose discontinuous gradient method was employed that yielded highly enriched mitochondrial populations free of myelin contamination. The activities of Complexes I, II, III, and II/III were lower in Syn than in NS mitochondria, while Complexes I/III and IV activities were similar in both populations. Shotgun lipidomics showed that levels of cardiolipin (Ptd(2)Gro) were lower, whereas levels of ceramide and phosphatidylserine were higher in Syn than in NS mitochondria. Coenzyme Q(9) and Q(10) was also lower in Syn than in NS mitochondria. Gangliosides, phosphatidic acid, sulfatides, and cerebrosides were undetectable in brain mitochondria. The distribution of Ptd(2)Gro molecular species was similar in both populations and formed a unique pattern, consisting of seven major molecular species groups, when arranged according to mass to charge ratios. Remodeling involving choline and ethanolamine phosphoglycerides could explain Ptd(2)Gro heterogeneity. NS and Syn mitochondrial lipidomic heterogeneity could influence energy metabolism, which may contribute to metabolic compartmentation of the brain.

Topics: Animals; Axons; Brain; Cardiolipins; Ceramides; Choline; Electron Transport Chain Complex Proteins; Energy Metabolism; Lipid Metabolism; Lipids; Mice; Mice, Inbred C57BL; Mitochondria; Phosphatidylserines; Presynaptic Terminals; Subcellular Fractions; Ubiquinone

Although many compounds have already been tested in-vitro to determine their antioxidant profile, it is necessary to investigate the in-vivo effect of potential antioxidants. However, representative models of systemic oxidative stress have been poorly studied. Here, different potential systemic oxidative stress animal models have been investigated. These included a vitamin E-deficient rat, a diabetic rat and an atherosclerotic rabbit model. Plasma/serum malondialdehyde was measured as a parameter of oxidative damage. Plasma/serum fat-soluble antioxidants were determined as markers of antioxidant defence. We demonstrated that vitamin E-deficient rats were not suitable as a model of systemic oxidative stress, whereas diabetic and atherosclerotic animals showed increased systemic oxidative damage, as reflected by significantly augmented plasma/serum malondialdehyde. Moreover, plasma coenzyme Q9 increased by 80% in diabetic rats, confirming systemic oxidative stress. In view of these observations and economically favouring factors, the diabetic rat appeared to be the most appropriate systemic oxidative stress model. These findings have provided important information concerning systemic oxidative stress animal models for the in-vivo study of antioxidants.

Topics: alpha-Tocopherol; Animals; Antioxidants; Atherosclerosis; Carbon Tetrachloride; Diabetes Mellitus, Experimental; gamma-Tocopherol; Lipid Peroxidation; Male; Malondialdehyde; Oxidative Stress; Rabbits; Rats; Rats, Sprague-Dawley; Rats, Wistar; Ubiquinone; Vitamin A; Vitamin E Deficiency

Coenzyme Q (Q) is a redox active lipid that is an essential component of the electron transport chain. Here, we show that steady state levels of Coq3, Coq4, Coq6, Coq7 and Coq9 polypeptides in yeast mitochondria are dependent on the expression of each of the other COQ genes. Submitochondrial localization studies indicate Coq9p is a peripheral membrane protein on the matrix side of the mitochondrial inner membrane. To investigate whether Coq9p is a component of a complex of Q-biosynthetic proteins, the native molecular mass of Coq9p was determined by Blue Native-PAGE. Coq9p was found to co-migrate with Coq3p and Coq4p at a molecular mass of approximately 1 MDa. A direct physical interaction was shown by the immunoprecipitation of HA-tagged Coq9 polypeptide with Coq4p, Coq5p, Coq6p and Coq7p. These findings, together with other work identifying Coq3p and Coq4p interactions, identify at least six Coq polypeptides in a multi-subunit Q biosynthetic complex.

Topics: Electrophoresis, Polyacrylamide Gel; Mitochondrial Membranes; Mitochondrial Proteins; Multienzyme Complexes; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins; Ubiquinone

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Female; Humans; Male; Middle Aged; New Zealand; Reference Values; Ubiquinone

Oxidative stress and antioxidants play an important role in neurodegenerative diseases. However, the exact participation of antioxidants in the evolution of prion diseases is still largely unknown. The aim of this study was to assess brain levels of coenzyme Q (CoQ), an endogenous lipophilic antioxidant, and the antioxidant/pro-oxidant status by determining oxidative damage to proteins and lipids after intracerebral bovine spongiform encephalopathy (BSE) infection of transgenic mice expressing bovine prion protein (PrP). Our results indicate that, whereas the ratio between the two CoQ homologues present in mice (CoQ(9) and CoQ(10)) is not altered by prion infection during the course of the disease, significant increases in total CoQ(9) and CoQ(10) were observed in BSE-infected mice 150 days after inoculation. This time point coincided with the first manifestation of PrP(Sc) deposition in nervous tissue. In addition, CoQ(9) and CoQ(10) levels, neuropathological alterations, and PrP(Sc) deposition in nervous tissues underwent further increases as the illness progressed. Lipid and protein oxidation were observed only at the final stage of the disease after clinical signs had appeared. These findings indicate upregulation of CoQ(9)- and CoQ(10)-dependent antioxidant systems in response to the increased oxidative stress induced by prion infection in nervous tissue. However, the induction of these endogenous antioxidant systems seems to be insufficient to prevent the development of the illness.

Topics: Animals; Antioxidants; Biomarkers; Brain; Cattle; Coenzymes; Disease Models, Animal; Encephalopathy, Bovine Spongiform; Lipid Metabolism; Mice; Mice, Transgenic; Oxidation-Reduction; Oxidative Stress; Prions; PrPSc Proteins; Ubiquinone; Up-Regulation

Dietary coenzyme Q(10) prolongs life span of rats fed on a PUFAn-6-enriched diet. Our aim was to analyze changes in the levels of plasma proteins of rats fed on a PUFAn-6 plus coenzyme Q(10)-based diet. This approach could give novel insights into the mechanisms of life span extension by dietary coenzyme Q(10) in the rat. Serum albumin, which decreases with aging in the rat, was significantly increased by coenzyme Q(10) supplementation both at 6 and 24 months. After depletion of the most abundant proteins by affinity chromatography, levels of less abundant plasma proteins were also studied by using 2D-electrophoresis and MALDI-TOF mass fingerprinting analysis. Our results have shown that lifelong dietary supplementation with coenzyme Q(10) induced significant decreases of plasma hemopexin, apolipoprotein H and inter-alpha-inhibitor H4P heavy chain (at both 6 and 24 months), preprohaptoglobin, fibrinogen gamma-chain precursor, and fetuin-like protein (at 6 months), and alpha-1-antitrypsin precursor and type II peroxiredoxin (at 24 months). On the other hand, coenzyme Q(10) supplementation resulted in significant increases of serine protease inhibitor 3, vitamin D-binding protein (at 6 months), and Apo A-I (at 24 months). Our results support a beneficial role of dietary coenzyme Q(10) decreasing oxidative stress and cardiovascular risk, and modulating inflammation during aging.

Topics: Animals; Blood Proteins; Cardiovascular Diseases; Coenzymes; Dietary Supplements; Electrophoresis, Gel, Two-Dimensional; Inflammation; Longevity; Male; Oxidative Stress; Proteome; Rats; Rats, Wistar; Ubiquinone

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Child; Child, Preschool; Chromatography, High Pressure Liquid; Coenzymes; Female; Humans; Infant; Infant, Newborn; Male; Middle Aged; Ubiquinone

Inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase (statins) have been proven to reduce effectively cholesterol level and morbidity and mortality in patients with coronary heart disease and/or dyslipoproteinemia. Statins inhibit synthesis of mevalonate, a precursor of both cholesterol and coenzyme Q (CoQ). Inhibited biosynthesis of CoQ may be involved in some undesirable actions of statins. We investigated the effect of simvastatin on tissue CoQ concentrations in the rat model of NO-deficient hypertension induced by chronic L-NAME administration. Male Wistar rats were treated daily for 6 weeks with L-NAME (40 mg/kg) or with simvastatin (10 mg/kg), another group received simultaneously L-NAME and simvastatin in the same doses. Coenzyme Q(9) and Q(10) concentrations were analyzed by high performance liquid chromatography. L-NAME and simvastatin alone had no effect on CoQ concentrations. However, simultaneous application of L-NAME and simvastatin significantly decreased concentrations of both CoQ homologues in the left ventricle and slightly decreased CoQ(9) concentration in the skeletal muscle. No effect was observed on CoQ level in the liver and brain. We conclude that the administration of simvastatin under the condition of NO-deficiency reduced the level of CoQ in the heart and skeletal muscle what may participate in adverse effect of statins under certain clinical conditions.

Topics: Animals; Brain; Coenzymes; Disease Models, Animal; Down-Regulation; Heart Ventricles; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Hypertension; Liver; Male; Muscle, Skeletal; NG-Nitroarginine Methyl Ester; Nitric Oxide; Rats; Rats, Wistar; Simvastatin; Time Factors; Ubiquinone

The SAMP1 strain is a mouse model for accelerated senescence and severe senile amyloidosis. We determined whether supplementation with coenzyme Q10 (CoQ10) could decelerate aging in SAMP1 mice and its potential role in aging. Plasma concentrations of CoQ10 and CoQ9 decreased with age in SAMP1 but not in SAMR1 mice. Supplementation with reduced CoQ10 (CoQH2, 250 mg/kg/day) for one week increased plasma CoQ10 concentrations, with an accompanying decrease in plasma CoQ9 concentrations. In two series of experiments, lifelong supplementation with CoQH2 decreased the senescence grading scores from 10 to 14 months, 7 to 15 months, and at 17 months of age. The body weight of female mice increased from 2 to 10 months of age versus controls in the second series of experiments. Lifelong CoQH2 supplementation did not prolong or shorten the lifespan, nor did it alter the murine senile amyloid (AApoAII) deposition rate or cancer incidence. In the second series of experiments, urinary levels of 8-hydroxydeoxyguanosine did not change with age or long-term supplementation with CoQH2. Urinary levels of acrolein (ACR)-lysine adduct increased significantly with age in SAMP1 mice; however, CoQH2 had no effect. Thus, lifelong dietary supplementation with CoQH2 decreased the degree of senescence in middle-aged SAMP1 mice.

Topics: Aging, Premature; Amyloidosis; Animals; Apolipoprotein A-II; Biomarkers; Coenzymes; Dietary Supplements; Female; Longevity; Male; Mice; Mice, Mutant Strains; Models, Animal; Oxidative Stress; Random Allocation; Ubiquinone

Coenzyme Q (CoQ), an electron transfer molecule in the respiratory chain and a lipid-soluble antioxidant, is present in almost all organisms. Most cereal crops produce CoQ9, which has nine isoprene units. CoQ10, with 10 isoprene units, is a very popular food supplement. Here, we report the genetic engineering of rice to produce CoQ10 using the gene for decaprenyl diphosphate synthase (DdsA). The production of CoQ9 was almost completely replaced with that of CoQ10, despite the presence of endogenous CoQ9 synthesis. DdsA designed to express at the mitochondria increased accumulation of total CoQ amount in seeds.

Topics: Alkyl and Aryl Transferases; Antioxidants; Base Sequence; Coenzymes; Crops, Agricultural; Electron Transport; Genetic Engineering; Mitochondria; Molecular Sequence Data; Oryza; Plants, Genetically Modified; Seeds; Terpenes; Ubiquinone

A procedure was developed to isolate fractions enriched in plasma membrane from Caenorhabditis elegans. Coenzyme Q9 (Q9) was found in plasma membrane isolated from either wild-type or long-lived qm30 and qm51 clk-1 mutant strains of Caenorhabditis elegans, along with dietary coenzyme Q8 (Q8) and the biosynthetic intermediate demethoxy-Q9 (DMQ9). NADH was able to reduce both Q8 and Q9, but not DMQ9. Our results indicate that DMQ9 cannot achieve the same redox role of Q9 in plasma membrane, suggesting that proportion of all these Q isoforms in plasma membrane must be an important factor in establishing the clk-1 mutant phenotype.

Topics: Animals; Caenorhabditis elegans; Caenorhabditis elegans Proteins; Cell Membrane; NAD; Oxidation-Reduction; Ubiquinone

There is substantial evidence that a bioenergetic defect may play a role in the pathogenesis of Huntington's Disease (HD). A potential therapy for remediating defective energy metabolism is the mitochondrial cofactor, coenzyme Q10 (CoQ10). We have reported that CoQ10 is neuroprotective in the R6/2 transgenic mouse model of HD. Based upon the encouraging results of the CARE-HD trial and recent evidence that high-dose CoQ10 slows the progressive functional decline in Parkinson's disease, we performed a dose ranging study administering high levels of CoQ10 from two commercial sources in R6/2 mice to determine enhanced efficacy. High dose CoQ10 significantly extended survival in R6/2 mice, the degree of which was dose- and source-dependent. CoQ10 resulted in a marked improvement in motor performance and grip strength, with a reduction in weight loss, brain atrophy, and huntingtin inclusions in treated R6/2 mice. Brain levels of CoQ10 and CoQ9 were significantly lower in R6/2 mice, in comparison to wild type littermate control mice. Oral administration of CoQ10 elevated CoQ10 plasma levels and significantly increased brain levels of CoQ9, CoQ10, and ATP in R6/2 mice, while reducing 8-hydroxy-2-deoxyguanosine concentrations, a marker of oxidative damage. We demonstrate that high-dose administration of CoQ10 exerts a greater therapeutic benefit in a dose dependent manner in R6/2 mice than previously reported and suggest that clinical trials using high dose CoQ10 in HD patients are warranted.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Adenosine Triphosphate; Animals; Body Weight; Coenzymes; Deoxyguanosine; Disease Models, Animal; Dose-Response Relationship, Drug; Huntingtin Protein; Huntington Disease; Male; Mice; Mice, Transgenic; Neostriatum; Nerve Tissue Proteins; Neuroprotective Agents; Nuclear Proteins; Rotarod Performance Test; Treatment Outcome; Ubiquinone

Coenzyme Q(10) (CoQ(10)), like other CoQs of various organisms, plays indispensable roles not only in energy generation but also in several other processes required for cells' survival. In this study, a gene encoding for a decaprenyl diphosphate synthase (Rsdds) was cloned from Rhodobacter sphaeroides in Escherichia coli. The in vivo catalytic activity and product specificity of Rsdds were compared with those of a counterpart enzyme from Agrobacterium tumefaciens (Atdds) in E. coli as a heterologous host. In contrast with Atdds, Rsdds showed lower catalytic activity but higher product specificity for CoQ(10) production, as indicated by the amount of CoQ(9) formation. The higher product specificity of Rsdds was also confirmed by utilizing both Rsdds and Atdds for in vitro synthesis of polyprenyl diphosphates. Thin layer chromatography indicated that the Rsdds enzyme resulted in relatively much less solanesyl diphosphate formation. The purified Rsdds catalyzed the addition of isopentenyl diphosphate to dimethyl allyl diphosphate, geranyl diphosphate, omega,E,E-farnesyl diphosphate (FPP), and omega,E,E,E-geranylgeranyl diphosphate as priming substrates. The kinetic parameters of V (max) (pmol/min), K (M) (microM), k (cat) (1/min), and k (cat) /K (M) of the enzyme using FPP as the most appropriate substrate were determined to be 264.6, 13.1, 8.8, and 0.67, respectively.

Topics: Agrobacterium tumefaciens; Alkyl and Aryl Transferases; Chromatography, Thin Layer; Cloning, Molecular; Coenzymes; Diphosphates; Diterpenes; DNA, Bacterial; Electrophoresis, Polyacrylamide Gel; Escherichia coli; Hemiterpenes; Kinetics; Molecular Sequence Data; Organophosphorus Compounds; Polyisoprenyl Phosphates; Rhodobacter sphaeroides; Sequence Analysis, DNA; Sesquiterpenes; Substrate Specificity; Ubiquinone

Coenzyme Q (CoQ) is an endogenous enzyme cofactor that may provide protective benefits as an antioxidant. In this study, in order to determine whether the concentrations of CoQ(9) are associated with the oxidative status in vivo, the effects of dietary supplements of CoQ(9) on mice were evaluated by using a new biomarker, total hydroxyoctadecadienoic acid (tHODE). Biological samples were first reduced and then saponified to convert the various oxidation products of linoleates to tHODE. Subsequently, by using GC-MS analyses, we simultaneously determined the absolute concentration of tHODE; its stereoisomer ratio, 9- and 13-(Z,E)-HODE/9- and 13-(E,E)-HODE, which is a measure of the hydrogen donor capacity of antioxidants; and the concentration of 8-iso-prostaglandin F(2alpha) (8-iso-PGF(2alpha)). Remarkable decreases in tHODE and 8-iso-PGF(2alpha) levels were observed in the plasma, erythrocytes, liver, and brain of mice that were maintained for 1 month on an alpha-tocopherol (alphaT)-free (E-free) diet supplemented with ubiquinone-9 (Q(9); 0.04 wt.%) as compared to those of mice that were fed an E-free diet. The (Z,E/E,E) HODE ratio was increased in the plasma and erythrocytes of mice that were fed a Q(9)-fortified diet as compared to those that were fed an E-free diet. In particular, the (Z,E/E,E) HODE ratios in the plasma and brain were significantly correlated with the concentrations of ubiquinol-9 (Q(9)H(2)). Further, the liver and brain levels of tHODE and 8-iso-PGF(2alpha) were significantly correlated with the plasma and erythrocyte levels of tHODE and 8-iso-PGF(2alpha), respectively, and in some cases, also exhibited significant correlations with antioxidants. These results indicate that the plasma and erythrocyte levels of tHODE and its stereoisomeric ratio can be prominent biomarkers for the evaluation of the oxidative status and antioxidant capacity in vivo, including in the liver and brain, and that CoQ plays a major role in the in vivo antioxidant network.

Topics: Alanine Transaminase; alpha-Tocopherol; Animals; Antioxidants; Brain Chemistry; Diet; Dinoprost; Fatty Acids, Unsaturated; Lipid Peroxidation; Liver; Male; Mice; Mice, Inbred C57BL; Oxidative Stress; Stereoisomerism; Ubiquinone

We report the cloning of a Trypanosoma cruzi gene encoding a solanesyl-diphosphate synthase, TcSPPS. The amino acid sequence (molecular mass approximately 39 kDa) is homologous to polyprenyl-diphosphate synthases from different organisms, showing the seven conserved motifs and the typical hydrophobic profile. TcSPPS preferred geranylgeranyl diphosphate as the allylic substrate. The final product, as determined by TLC, had nine isoprene units. This suggests that the parasite synthesizes mainly ubiquinone-9 (UQ-9), as described for Trypanosoma brucei and Leishmania major. In fact, that was the length of the ubiquinone extracted from epimastigotes, as determined by high-performance liquid chromatography. Expression of TcSPPS was able to complement an Escherichia coli ispB mutant. A punctuated pattern in the cytoplasm of the parasite was detected by immunofluorescence analysis with a specific polyclonal antibody against TcSPPS. An overlapping fluorescence pattern was observed using an antibody directed against the glycosomal marker pyruvate phosphate dikinase, suggesting that this step of the isoprenoid biosynthetic pathway is located in the glycosomes. Co-localization in glycosomes was confirmed by immunogold electron microscopy and subcellular fractionation. Because UQ has a central role in energy production and in reoxidation of reduction equivalents, TcSPPS is promising as a new chemotherapeutic target.

Topics: Alkyl and Aryl Transferases; Amino Acid Sequence; Animals; Chromatography, Thin Layer; Cloning, Molecular; Cosmids; Escherichia coli; Genetic Complementation Test; Microbodies; Mitochondria; Molecular Sequence Data; Sequence Homology, Amino Acid; Substrate Specificity; Trypanosoma cruzi; Ubiquinone

We investigated the application of 1-alkylamines, as additives to the mobile phase, to a quantification method for ubiquinone-9 (CoQ9) and ubiquinone-10 (CoQ10) in rat thigh muscle and heart using liquid chromatography-tandem mass spectrometry (LC-MS/MS). In the optimization of the analytical method, we found that 1-alkylamines mixed with CoQ9 and CoQ10 in the turbo ion sprayed solution formed the 1-alkylammonium adduct molecules of these compounds during the ionization process and that the intensity of the adduct ions was considerably higher than that of the protonated molecules ([M+H]+) of these compounds. Furthermore, we investigated a variety of 1-alkylamines in the mobile phase for LC-MS/MS analysis to select the most appropriate 1-alkylamine for higher sensitivities of CoQ9 and CoQ10. After these examinations, we found that methylamine was the most suitable additive for the mobile phase, allowing a 12.5-fold gain in signal intensity in the full ion mass spectrum compared with that without methylamine. The internal standard (IS) used was ubiquinone-11 (CoQ11) for each analyte. The analytes and IS were extracted with methanol from the tissue homogenates at neutral pH and were injected into an LC-MS/MS with a turbo ion spray interface. The calibration curves for CoQ9 (5-500 microg/g in thigh muscle and 50-10,000 microg/g in heart) and CoQ10 (1-500 microg/g in thigh muscle and 10-10,000 microg/g in heart) showed good linearity. The method was precise; the relative standard deviations of the method for rat thigh muscle were not more than 13.5 and 9.0% for CoQ9 and CoQ10, respectively, and those for rat heart were not more than 6.7 and 5.4% for CoQ9 and CoQ10, respectively. The accuracies of the method for both rat thigh muscle and heart were good, with the deviations between the nominal concentration and calculated concentration of CoQ9 and CoQ10 typically being within 12.3 and 4.3%, respectively. This method provided reliable concentration levels for CoQ9 and CoQ10 in rat thigh muscle and heart.

Topics: Amines; Animals; Chromatography, High Pressure Liquid; Gas Chromatography-Mass Spectrometry; Male; Methylamines; Muscles; Myocardium; Rats; Reproducibility of Results; Sensitivity and Specificity; Ubiquinone

The structure of the Blastochloris viridis photosynthetic reaction center has been determined at 100 K by flash-freezing crystals. A data set to 2.2 A resolution provides a well determined model of the wild-type protein. Of particular interest are the position, occupancy and heterogeneity of the Q(B)-binding site. Data were also collected from a crystal frozen immediately after illumination. The data support predominant binding of Q(B) in the proximal position in both the neutral and charge-separated states.

Topics: Binding Sites; Cryoprotective Agents; Crystallization; Darkness; Freezing; Herbicides; Light; Models, Molecular; Photosynthesis; Proteobacteria; Quinones; Ubiquinone; Water

Monitoring in vivo oxidative stress implicates the evaluation of damage and defence parameters by well-established, validated methods. We report two optimized and validated HPLC methods for measurement of malondialdehyde (MDA) and fat-soluble vitamins in rat plasma. For the MDA method, optimization experiments of the thiobarbituric acid test resulted in the addition of 1% butylhydroxytoluene to the reaction mixture and in a heating time reduction to 40 min, ensuring inhibition of further lipid peroxidation during the test. Validation experiments showed good linearity, precision and recovery. The use of HPLC with coulometric array detection technology permits simultaneous and sensitive analysis of different fat-soluble vitamins and related compounds (tocopherols, retinoids, carotenoids and coenzyme Q10), which are identified by both retention time and electrochemical characteristics. Furthermore, this method is extended to the analysis of coenzyme Q9, the predominant homologue in rats. Validation experiments with rat plasma gave good results.

Topics: Animals; Chromatography, High Pressure Liquid; Fats; Lipid Peroxidation; Male; Malondialdehyde; Oxidative Stress; Rats; Rats, Sprague-Dawley; Solubility; Tocopherols; Ubiquinone; Vitamin A; Vitamins

Currently, eight genes are known to be involved in coenzyme Q6 biosynthesis in Saccharomyces cerevisiae. Here, we report a new gene designated COQ9 that is also required for the biosynthesis of this lipoid quinone. The respiratory-deficient pet mutant C92 was found to be deficient in coenzyme Q and to have low mitochondrial NADH-cytochrome c reductase activity, which could be restored by addition of coenzyme Q2. The mutant was used to clone COQ9, corresponding to reading frame YLR201c on chromosome XII. The respiratory defect of C92 is complemented by COQ9 and suppressed by COQ8/ABC1. The latter gene has been shown to be required for coenzyme Q biosynthesis in yeast and bacteria. Suppression by COQ8/ABC1 of C92, but not other coq9 mutants tested, has been related to an increase in the mitochondrial concentration of several enzymes of the pathway. Coq9p may either catalyze a reaction in the coenzyme Q biosynthetic pathway or have a regulatory role similar to that proposed for Coq8p.

Topics: Chromatography, High Pressure Liquid; Cloning, Molecular; Cytochromes; Mutation; NAD; Oxidoreductases; Phenotype; Saccharomyces cerevisiae; Spectrophotometry; Ubiquinone

Coenzyme Q(10) supplementation on age-related changes in oxidative stress and function of heart mitochondria in rats fed a polyunsaturated fatty acid (PUFA)-rich diet was investigated. Two groups of rats were fed for 24 months on a PUFA-rich diet, differing in supplementation or not with coenzyme Q(10). Animals were killed at 6, 12, or 24 months. Fatty-acid profile, hydroperoxides, alpha-tocopherol, coenzyme Q, catalase and glutathione peroxidase activities, and cytochromes a+a(3), b, c+c(1) and cytochrome c oxidase activity were measured. Coenzyme Q(10)-supplemented animals showed lower hydroperoxide levels; higher content and/or activity of alpha-tocopherol, coenzyme Q, and catalase; and a slightly lower decrease in mitochondrial function. According to that, previously reported positive effects of coenzyme Q supplementation on the life span of rats fed a PUFA-rich diet might be a consequence, at least in part, of a lower oxidative stress level and perhaps, to a minor extent, of a smaller decrease in mitochondrial function.

Topics: Aging; alpha-Tocopherol; Animals; Catalase; Coenzymes; Dietary Fats, Unsaturated; Electron Transport Complex IV; Fatty Acids, Unsaturated; Glutathione Peroxidase; Lipid Peroxides; Male; Mitochondria, Heart; Oxidative Stress; Rats; Rats, Wistar; Ubiquinone

Life-long low-dosage supplementation of coenzyme Q(10) (CoQ(10)) is studied in relation to the antioxidant status and DNA damage. Thirty-two male rats were assigned into two experimental groups differing in the supplementation or not with 0.7 mg/kg/day of CoQ(10). Eight rats per group were killed at 6 and 24 months. Plasma retinol, alpha-tocopherol, coenzyme Q, total antioxidant capacity and fatty acids were analysed. DNA strand breaks were studied in peripheral blood lymphocytes. Aging and supplementation led to significantly higher values for CoQ homologues, retinol and alpha-tocopherol. No difference in total antioxidant capacity was detected at 6 months but significantly lower values were found in aged control animals. Similar DNA strand breaks levels were found at 6 months. Aging led to significantly higher DNA strand breaks levels in both groups but animals supplemented with CoQ(10) led to a significantly lower increase in that marker. Aged rats showed significantly higher polyunsaturated fatty acids. This study demonstrates that lifelong intake of a low dosage of CoQ(10) enhances plasma levels of CoQ(9), CoQ(10), alpha-tocopherol and retinol. In addition, CoQ(10) supplementation attenuates the age-related fall in total antioxidant capacity of plasma and the increase in DNA damage in peripheral blood lymphocytes.

Topics: Aging; alpha-Tocopherol; Animals; Antioxidants; Coenzymes; DNA Damage; Fatty Acids; Fatty Acids, Monounsaturated; Fatty Acids, Unsaturated; Male; Rats; Rats, Wistar; Ubiquinone; Vitamin A

Green tea (Camellia sinensis), and CoQ(9 )when given to Wistar rats produced a partial reversal on reserpine induced oxidative stress and liver damage. Green tea, with its abundant polyphenol (-)Epigallocatechin 3-gallate (ECGC) and other catechins, is known for its antioxidative characteristics influencing lipid metabolism. Ubiquinone, abundant in heart muscle, is also a potent antioxidant with known effects in numerous pathologies. However the combined effect of ECGC and ubiquninone has not been reported. In the present study we found that green tea extract, when given in combination with CoQ(9) to Wistar rats subjected to oxidative stress, showed a statistically significant antioxidative effect. Liver cholesterol level in rats receiving combination treatment was also significantly lower than control or rats receiving green tea extract alone. Reserpine induced liver damage in Wistar rats was also partially reversed by a treatment of green tea extract when combined with CoQ(9). These results may have important clinical implications and may be extrapolated for the treatment of patients suffering from liver damage due to hepatitis B/C or liver cirrhosis.

Topics: Alanine Transaminase; Animals; Antioxidants; Aspartate Aminotransferases; Camellia sinensis; Cholesterol; Drug Synergism; Liver; Male; Oxidative Stress; Plant Extracts; Rats; Rats, Wistar; Reserpine; Tea; Thiobarbituric Acid Reactive Substances; Ubiquinone

3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors have been shown to prevent or reverse hypertrophy of the LV in several models of left ventricular hypertrophy. The aim of the present study was to determine whether treatment with simvastatin can prevent hypertension, reduction of tissue nitric oxide synthase activity and left ventricular (LV) remodeling in NG-nitro-L-arginine methyl ester(L-NAME)-induced hypertension. Four groups of rats were investigated: control, simvastatin (10 mg/kg), L-NAME (40 mg/kg) and L-NAME + simvastatin (in corresponding doses). Animals were sacrificed and studied after 6 weeks of treatment. The decrease of NO-synthase activity in the LV, kidney and brain was associated with hypertension, LV hypertrophy and fibrosis development and remodeling of the aorta in the L-NAME group. Simvastatin attenuated the inhibition of NO-synthase activity in kidney and brain, partly prevented hypertension development and reduced the concentration of coenzyme Q in the LV. Nevertheless, myocardial hypertrophy, fibrosis and enhancement of DNA concentration in the LV, and remodeling of the aorta were not prevented by simultaneous simvastatin treatment in the L-NAME treated animals. We conclude that the HMG-CoA reductase inhibitor simvastatin improved nitric oxide production and partially prevented hypertension development, without preventing remodeling of the left ventricle and aorta in NO-deficient hypertension.

Topics: Animals; Aorta; Blood Pressure; Body Weight; Coenzymes; DNA; Enzyme Inhibitors; Fibrosis; Hemodynamics; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Hypertension; Male; Myocardium; NG-Nitroarginine Methyl Ester; Nitric Oxide Synthase; Nitric Oxide Synthase Type III; Organ Size; Rats; Rats, Wistar; Simvastatin; Ubiquinone; Ventricular Remodeling

Ubiquinone-responsive multiple respiratory chain dysfunction due to coenzyme Q(10) (CoQ(10)) deficiency has been previously identified in muscle biopsies. However, previous methods are unreliable for estimating CoQ(10) redox status in tissue. We developed an accurate method for measuring tissue concentrations of reduced and oxidized coenzyme Q (CoQ).. Mouse tissues were weighed in the frozen state and homogenized with cold 1-propanol on ice. After solvent extraction, centrifugation and filtration, the filtrate was subsequently analyzed by reversed-phase HPLC with coulometric detection.. Reference calibration curves were used to determine reduced and oxidized coenzyme Q(9) (CoQ(9)) and CoQ(10) concentrations in tissues. The method is sensitive ( approximately 15 microg/l), reproducible (6% CV) for CoQ(9) and CoQ(10), and linear up to 20 mg/l for CoQ(9) and CoQ(10). Analytical recoveries were 90-104%. In mouse tissues the amounts of total CoQ (TQ) ranged from 261 to 1737 nmol/g of protein. Total CoQ(9) levels are comparable with the values of those previously reported. CoQ is found to be mostly in the reduced form in mouse liver ( approximately 87%), heart ( approximately 60%), and muscle tissues ( approximately 58%); in the brain, most of the CoQ is in the oxidized state ( approximately 65%).. This procedure provides a precise, sensitive, and direct assay method for the determination of reduced and oxidized CoQ(9) and CoQ(10) in mouse hindleg muscle, heart, brain, and liver tissues.

Topics: Animals; Calibration; Chromatography, High Pressure Liquid; Coenzymes; Electrochemistry; Mice; Mice, Inbred C57BL; Oxidation-Reduction; Reference Standards; Reproducibility of Results; Tissue Distribution; Ubiquinone

Previous studies have shown that T3 treatment and cold exposure induce similar biochemical changes predisposing rat liver to oxidative stress. This suggests that the liver oxidative damage observed in experimental and functional hyperthyroidism is mediated by thyroid hormone. To support this hypothesis we investigated whether middle-term cold exposure (2 and 10 days), like T3 treatment, also increases H2O2 release by liver mitochondria. We found that the rate of H2O2 release increased only during State 4 respiration, but faster flow of reactive oxygen species (ROS) from mitochondria to the cytosolic compartment was ensured by the concomitant increase in tissue mitochondrial proteins. Cold exposure also increased the capacity of mitochondria to remove H2O2. This indicates that cold causes accelerated H2O2 production, which might depend on enhanced autoxidizable carrier content and should lead to increased mitochondrial damage. Accordingly, mitochondrial levels of hydroperoxides and protein-bound carbonyls were higher after cold exposure. Levels of low-molecular weight antioxidants were not related to the extent of oxidative damage, but susceptibility to both in vitro oxidative challenge and Ca2+-induced swelling increased in mitochondria from cold exposed rats. The cold-induced changes in several parameters, including susceptibility to swelling, were time dependent, because they were apparent or greater after 10 days cold exposure. The cold-induced increase in swelling may be a feedback mechanism to limit tissue oxidative stress, purifying the mitochondrial population from ROS-overproducing mitochondria, and the time course for such change is consistent with the gradual development of cold adaptation.

Topics: Animals; Antioxidants; Cold Temperature; Electron Transport Complex IV; Hydrogen Peroxide; Hyperthyroidism; Lipid Peroxidation; Male; Mitochondria, Liver; Mitochondrial Swelling; Oxidative Stress; Oxygen Consumption; Rats; Rats, Wistar; Thyroid Gland; Time Factors; Ubiquinone; Uncoupling Agents; Vitamin E

Administration of dehydroepiandrosterone sulfate (DHEAS) for 14 days caused a significant increase in the total ubiquinone-9 level in the hepatic tissue of male F-344 rats (p < 0.05). Hepatic ubiquinone-10 level of DHEAS-treated rats however, was found not to be statistically different from control animals. These findings suggest that peroxisome proliferator DHEAS displays typical hepatic response in increasing ubiquinone concentration in the rat.

Topics: Animals; Antioxidants; Dehydroepiandrosterone Sulfate; Liver; Male; Oxidative Stress; Rats; Rats, Inbred F344; Stimulation, Chemical; Ubiquinone

The long-lived mutant of Caenorhabditis elegans, clk-1, is unable to synthesize ubiquinone, CoQ(9). Instead, the mutant accumulates demethoxyubiquinone(9) and small amounts of rhodoquinone(9) as well as dietary CoQ(8). We found a profound defect in oxidative phosphorylation, a test of integrated mitochondrial function, in clk-1 mitochondria fueled by NADH-linked electron donors, i.e. complex I-dependent substrates. Electron transfer from complex I to complex III, which requires quinones, is severely depressed, whereas the individual complexes are fully active. In contrast, oxidative phosphorylation initiated through complex II, which also requires quinones, is completely normal. Here we show that complexes I and II differ in their ability to use the quinone pool in clk-1. This is the first direct demonstration of a differential interaction of complex I and complex II with the endogenous quinone pool. This study uses the combined power of molecular genetics and biochemistry to highlight the role of quinones in mitochondrial function and aging.

Topics: Animals; Ascorbic Acid; Caenorhabditis elegans; Electron Transport Complex I; Electron Transport Complex II; Glutamic Acid; Hydroquinones; Malates; Mitochondria; Mutation; Oxidative Phosphorylation; Pyruvic Acid; Quinones; Substrate Specificity; Tetramethylphenylenediamine; Time Factors; Ubiquinone

Mitochondria-produced reactive oxygen species (ROS) are thought to contribute to cell death caused by a multitude of pathological conditions. The molecular sites of mitochondrial ROS production are not well established but are generally thought to be located in complex I and complex III of the electron transport chain. We measured H(2)O(2) production, respiration, and NADPH reduction level in rat brain mitochondria oxidizing a variety of respiratory substrates. Under conditions of maximum respiration induced with either ADP or carbonyl cyanide p-trifluoromethoxyphenylhydrazone,alpha-ketoglutarate supported the highest rate of H(2)O(2) production. In the absence of ADP or in the presence of rotenone, H(2)O(2) production rates correlated with the reduction level of mitochondrial NADPH with various substrates, with the exception of alpha-ketoglutarate. Isolated mitochondrial alpha-ketoglutarate dehydrogenase (KGDHC) and pyruvate dehydrogenase (PDHC) complexes produced superoxide and H(2)O(2). NAD(+) inhibited ROS production by the isolated enzymes and by permeabilized mitochondria. We also measured H(2)O(2) production by brain mitochondria isolated from heterozygous knock-out mice deficient in dihydrolipoyl dehydrogenase (Dld). Although this enzyme is a part of both KGDHC and PDHC, there was greater impairment of KGDHC activity in Dld-deficient mitochondria. These mitochondria also produced significantly less H(2)O(2) than mitochondria isolated from their littermate wild-type mice. The data strongly indicate that KGDHC is a primary site of ROS production in normally functioning mitochondria.

Topics: Adenosine Diphosphate; Animals; Antimycin A; Coenzymes; Dihydrolipoamide Dehydrogenase; Electron Transport; Electron Transport Complex I; Hydrogen Peroxide; Intracellular Membranes; Ketoglutarate Dehydrogenase Complex; Ketoglutaric Acids; Membrane Potentials; Mice; Mice, Knockout; Mitochondria; NAD; NADP; Nerve Tissue Proteins; Oligomycins; Oxidation-Reduction; Prosencephalon; Pyruvate Dehydrogenase Complex; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Rotenone; Succinic Acid; Superoxide Dismutase; Superoxides; Ubiquinone

Three yellow-pigmented, Gram-negative, rod-shaped, non-spore-forming bacterial strains, C36T, C37 and C39, were isolated in the Medical Clinic for Small Animals and Ungulates at the University for Veterinary Medicine in Vienna, Austria. On the basis of 16S rRNA gene sequence similarity, strain C36T was shown to belong to the genus Pseudomonas; Pseudomonas oleovorans DSM 1045T was the nearest relative (99.5 % sequence similarity). Other Pseudomonas species shared <97 % sequence similarity with strain C36T. The presence of Q-9 as the major ubiquinone, the predominance of putrescine and spermidine in its polyamine patterns and its fatty acid profile [i.e. the predominance of C(16 : 0), summed feature 3 (C(16 : 1)omega7c and/or 2-OH C(15 : 0) iso), C(18 : 1)omega7c and the presence of 3-OH C(10 : 0), 3-OH C(12 : 0) and 2-OH C(12 : 0)] were in agreement with identification of this strain as a member of the genus Pseudomonas. Physiological and biochemical characteristics and the results of genomic fingerprinting clearly differentiated strain C36T from its phylogenetic relative P. oleovorans DSM 1045T. Results from DNA-DNA hybridization showed that strain C36T represents a species that is distinct from P. oleovorans DSM 1045T. These data demonstrate that strain C36T represents a novel species of the genus Pseudomonas, for which the name Pseudomonas psychrotolerans sp. nov. is proposed. The type strain is C36T (= LMG 21977T = DSM 15758T). Additionally, physiological, biochemical, chemotaxonomic and genomic fingerprints indicate that P. oleovorans ATCC 29347 may not be a member of the species P. oleovorans sensu stricto.

Topics: Animals; Austria; Bacterial Typing Techniques; DNA Fingerprinting; DNA, Bacterial; DNA, Ribosomal; Fatty Acids; Genes, rRNA; Molecular Sequence Data; Nucleic Acid Hybridization; Phylogeny; Pigments, Biological; Pseudomonas; Pseudomonas oleovorans; Putrescine; RNA, Bacterial; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Spermidine; Ubiquinone

The effect of rooibos tea (Aspalathus linearis) on liver antioxidant status and oxidative stress was investigated in rat model of carbon tetrachloride-induced liver damage. Synthetic antioxidant N-acetyl-L-cysteine (NAC) was used for comparison. Administration of carbon tetrachloride (CCl4) for 10 weeks decreased liver concentrations of reduced and oxidized forms of coenzyme Q9 (CoQ9H2 and CoQ9), reduced -tocopherol content and simultaneously increased the formation of malondialdehyde (MDA) as indicator of lipid peroxidation. Rooibos tea and NAC administered to CCl4-damaged rats restored liver concentrations of CoQ9H2 and alpha-tocopherol and inhibited the formation of MDA, all to the values comparable with healthy animals. Rooibos tea did not counteract the decrease in CoQ9, whereas NAC was able to do it. Improved regeneration of coenzyme Q9 redox state and inhibition of oxidative stress in CCl4-damaged livers may explain the beneficial effect of antioxidant therapy. Therefore, the consumption of rooibos tea as a rich source of natural antioxidants could be recommended as a market available, safe and effective hepatoprotector in patients with liver diseases.

Topics: Acetylcysteine; Animals; Antioxidants; Aspalathus; Beverages; Carbon Tetrachloride; Liver Failure, Acute; Liver Regeneration; Male; Oxidation-Reduction; Oxidative Stress; Phytotherapy; Plant Extracts; Rats; Rats, Wistar; Treatment Outcome; Ubiquinone

Topics: Antioxidants; Chromatography, High Pressure Liquid; Humans; Indicators and Reagents; Oxidation-Reduction; Plasma; Reproducibility of Results; Ubiquinone

Two solanesyl diphosphate synthases, designated SPS1 and SPS2, which are responsible for the synthesis of the isoprenoid side chain of either plastoquinone or ubiquinone in Arabidopsis thaliana, were identified. Heterologous expression of either SPS1 or SPS2 allowed the generation of UQ-9 in a decaprenyl diphosphate synthase-defective strain of fission yeast and also in wild-type Escherichia coli. SPS1-GFP was found to localize in the ER while SPS2-GFP localized in the plastid of tobacco BY-2 cells. These two different subcellular localizations are thought to be the reflection of their roles in solanesyl diphosphate synthesis in two different parts: presumably SPS1 and SPS2 for the side chains of ubiquinone and plastoquinone, respectively.

Topics: Alkyl and Aryl Transferases; Arabidopsis; Arabidopsis Proteins; DNA, Complementary; Endoplasmic Reticulum; Escherichia; Molecular Sequence Data; Nicotiana; Plastids; Plastoquinone; Sequence Homology, Amino Acid; Sequence Homology, Nucleic Acid; Terpenes; Ubiquinone; Yeasts

Ubiquinone (coenzyme Q; Q) is a key factor in the mitochondria electron transport chain, but it also functions as an antioxidant and as a cofactor of mitochondrial uncoupling proteins. Furthermore, Q isoforms balance in Caenorhabditis elegans is determined by both dietary intake and endogenous biosynthesis. In the absence of synthesis, withdrawal of dietary Q8 in adulthood extends life span. Thus, Q plays an important role in the aging process and understanding its synthesis acquires a new impetus. We have identified by RNA interference (RNAi) eight genes, including clk-1, involved in ubiquinone biosynthesis in C. elegans feeding animals with dsRNA-containing Escherichia coli HT115 strains. Silenced C. elegans showed lower levels of both endogenous Q9 and Q8 provided by diet, produced less superoxide without a significant modification of mitochondrial electron chain, and extended life span compared with non-interfered animals. E. coli strains harboring dsRNA also interfered with their own Q8 biosynthesis. These findings suggest that more efficient electron transport between a lower amount of Q and electron transport capacity of the mitochondrial complexes leads to less production of reactive oxygen species that contributes to extension of life span in the nematode C. elegans.

Topics: Animals; Caenorhabditis elegans; Electron Transport; Escherichia coli; Longevity; Mitochondria; Models, Biological; RNA Interference; Superoxides; Transformation, Bacterial; Ubiquinone

Diabetes, which causes enhanced oxidative stress, is a multifactorial disease that leads to deleterious effects in many organ systems within the body. Ubiquinones (coenzyme Q(9) and Q(10)) are amphipathic molecular components of the electron transport chain that function also as endogenous antioxidants and attenuate the diabetes-induced decreases in antioxidant defense mechanisms. Insulin-like growth factor 1 (IGF-1) is considered to be an "essential surviving factor", the level and function of which are compromised in diabetes. This study investigated the impact of IGF-1 supplementation on ubiquinone levels in a rat model of type I diabetes. Adult male Sprague-Dawley rats were divided into four groups: control, control plus IGF-1, diabetic and diabetic plus IGF-1. Diabetic animals received a single intravenous injection of streptozotocin (STZ, 55 mg/kg). IGF-1 supplementation groups received a daily intraperitoneal dose of 3 mg IGF-1 per kilogram body weight for 7 weeks. Coenzyme Q(9) and Q(10) levels were assessed by ultraviolet detection on high pressure liquid chromatography. STZ caused a significant reduction in body weight and an elevation in blood glucose level, which were not prevented by IGF-1 supplementation. In addition Q(9) and Q(10) levels in diabetic liver were significantly elevated. IGF-1 supplementation prevented liver alterations in Q(10) but not Q(9) levels. Q(9) and Q(10) levels in diabetic kidney were significantly depressed, and these deleterious effects were abolished by IGF-1 treatment. These data suggest that IGF-1 antagonizes the diabetes-induced alterations in endogenous antioxidants including coenzyme Q(10), and hence may have a therapeutic role in diabetes.

Topics: Animals; Coenzymes; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 1; Dietary Supplements; Disease Models, Animal; Insulin-Like Growth Factor I; Kidney; Liver; Male; Oxidative Stress; Rats; Rats, Sprague-Dawley; Reference Values; Ubiquinone

The effects of the thyroid state on oxidative damage, antioxidant capacity, susceptibility to in vitro oxidative stress and Ca(2+)-induced permeabilization of mitochondria from rat tissues (liver, heart, and gastrocnemious muscle) were examined. Hypothyroidism was induced by administering methimazole in drinking water for 15 d. Hyperthyroidism was elicited by a 10 d treatment of hypothyroid rats with triiodothyronine (10 micro g/100 g body weight). Mitochondrial levels of hydroperoxides and protein-bound carbonyls significantly decreased in hypothyroid tissues and were reported above euthroid values in hypothyroid rats after T(3) treatment. Mitochondrial vitamin E levels were not affected by changes of animal thyroid state. Mitochondrial Coenzyme Q9 levels decreased in liver and heart from hypothyroid rats and increased in all hyperthyroid tissues, while Coenzyme Q10 levels decreased in hypothyroid liver and increased in all hyperthyroid tissues. The antioxidant capacity of mitochondria was not significantly different in hypothyroid and euthyroid tissues, whereas it decreased in the hyperthyroid ones. Susceptibility to in vitro oxidative challenge decreased in mitochondria from hypothyroid tissues and increased in mitochondria from hyperthyroid tissues, while susceptibility to Ca(2+)-induced swelling decreased only in hypothyroid liver mitochondria and increased in mitochondria from all hyperthyroid tissues. The tissue-dependence of the mitochondrial susceptibility to stressful conditions in altered thyroid states can be explained by different thyroid hormone-induced changes in mitochondrial ROS production and relative amounts of mitochondrial hemoproteins and antioxidants. We suggest that susceptibilities to oxidants and Ca(2+)-induced swelling may have important implications for the thyroid hormone regulation of the turnover of proteins and whole mitochondria, respectively.

Topics: Animals; Antioxidants; Calcium; Hyperthyroidism; Hypothyroidism; In Vitro Techniques; Lipid Peroxidation; Male; Methimazole; Mitochondria, Heart; Mitochondria, Liver; Mitochondria, Muscle; Mitochondrial Swelling; Oxidative Stress; Rats; Rats, Wistar; Triiodothyronine; Ubiquinone; Vitamin E

Cyclosporine A (CsA) plays a pivotal role in controlling Ca2+ movement in the cell modulating also the mitochondrial permeability transition pore. We investigated if chronic administration of CsA may have some effects on the lipophilic and hydrophilic antioxidant pattern of rat liver mitochondria and on their morphological structure. It seems that CsA administration does not statistically affect the redox status of the antioxidants investigated and their amounts (vitamin E, CoQ9, CoQ10, glutathione, uric acid and ascorbic acid) despite the variety of effects that this treatment produces at physiological and morphological levels. However, some kind of derangement could occur in the liver biochemical machinery since CsA treatment induces a markedly increased variability in antioxidant contents.

Topics: Animals; Antioxidants; Ascorbic Acid; Coenzymes; Cyclosporine; Glutathione; Male; Microscopy, Electron; Mitochondria, Liver; Rats; Rats, Wistar; Ubiquinone; Uric Acid; Vitamin E

Hypopituitary dwarf mice exhibit a heightened antioxidative capacity and live extensively longer than age-matched controls. Importantly, dwarf mice resist peripheral oxidative stress induced by paraquat, and behaviorally, they maintain cognitive function and locomotor activity at levels above those observed in old wild-type animals. We assessed monoaminergic neurotransmitters in nigrostriatal tract and cerebellum after the administration of the dopaminergic neurotoxin, MPTP. There was no significant change in mitochondrial monoamine oxidase (MAO)-B and total MAO activity in the substantia nigra and nucleus caudatus putamen of wild-type and dwarf mice. Coenzymes Q-9 and Q-10 were present in similar quantities, as were dopamine, norepinephrine, and serotonin levels in the cerebellum and nigrostriatal tract. MPTP set off tremor, hind limb abduction, and straub tail behavior and induced significant dopamine depletion in the striatum of both dwarf and normal mice. This study shows that the MAO activity and the coenzyme content of dwarf mice are similar to those of their wild-type controls and hence susceptible to MPTP-induced toxicity.

Topics: 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; Animals; Behavior, Animal; Caudate Nucleus; Coenzymes; Dopamine; Mice; Mice, Mutant Strains; Monoamine Oxidase; Oxidative Stress; Substantia Nigra; Ubiquinone

We investigated the effect of hyperthyroidism on the functional response of mitochondria to ischemia-reperfusion and its relationship with changes in mitochondrial susceptibility to stress conditions.. Hyperthyroidism was elicited by ten daily intraperitoneal injections of T3 (10 microg/100 g body weight). Mitochondria were isolated at 3000xg (M3) from homogenates of hearts perfused by the Langendorff technique after either 25 min reperfusion following 20 min ischemia or 45 min perfusion (controls). Rates of O2 consumption and H2O2 release with complex II-linked substrate, capacity to remove H2O2, extent of oxidative damage, levels of liposoluble antioxidants, such as ubiquinols and vitamin E, and susceptibility to Ca2+ -induced swelling were determined.. During reperfusion, hyperthyroid hearts displayed a significant tachycardia together with a low functional recovery. In comparison to the respective controls, mitochondria from both euthyroid and hyperthyroid hearts subjected to ischemia-reperfusion protocol exhibited decreases in the rate of O2 consumption, capacity to remove H2O2, and concentration of antioxidants, and increases in the rate of H2O2 release, concentration of hydroperoxides and protein-bound carbonyls, and susceptibility to Ca2+ -induced swelling. Such changes were higher in mitochondria from hyperthyroid hearts. The increase in the protein percent content and cytochrome oxidase activity of a mitochondrial fraction isolated at 8000xg (M8) from hyperthyroid hearts after reperfusion, suggests that the decline of mitochondrial respiration of M3 fraction could be due to the degradation of the oldest, mature mitochondria endowed of high oxidative capacity, but low antioxidant capacity, which would be lost by heavy mitochondrial fraction and recovered in the light fraction.. The higher susceptibility to ischemia-reperfusion of the heart from hyperthyroid animals is associated with a significant increase in mitochondrial dysfunction.

Topics: Animals; Antioxidants; Calcium; Coenzymes; Disease Susceptibility; Heart Rate; Hydrogen Peroxide; Hyperthyroidism; Male; Membrane Potentials; Mitochondria, Heart; Myocardial Reperfusion Injury; Oxygen Consumption; Rats; Rats, Wistar; Triiodothyronine; Ubiquinone; Vitamin E

A bacterial isolate, with an optimum growth temperature of about 50 degrees C, was recovered from the hot spring at Egerszalók in Hungary. Phylogenetic analyses using the 16S rRNA gene sequence of strain H-8T indicated that the new organism represented a new genus and species of alpha-1 subclass of the Proteobacteria. The major fatty acids of strain H-8T are 16:0, 18:1 omega7c; the rare fatty acid 19:0 20H cyclo 11,12 is also present. Ubiquinone 9 is the major respiratory quinone, the polar lipids are phosphatidylcholine, phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylglycerol in addition to two unidentified aminolipids. The new isolate forms red-colored colonies, flocculates in liquid media, is heterotrophic and strictly aerobic. Thiosulfate is oxidized to sulfate, but an increase in biomass could not be measured because of the flocculating behavior. Bacteriochloropyll a was detected by direct spectrophotometric analysis when the organism was grown at 30 degrees C, but could not be detected after growth at 50 degrees C. pufL and pufM genes were present. Heterotrophic growth of strain H-8T occurs on a few carbohydrates, amino acids and organic acids. On the basis of the phylogenetic analyses, physiological and biochemical characteristics, we propose that strain H-8T represents a new genus and a new species most closely related to Roseococcus thiosulfatophilus for which we propose the name Rubritepida flocculans.

Topics: Alphaproteobacteria; Amino Acid Sequence; Base Composition; Fatty Acids; Genes, Bacterial; Hydrogen-Ion Concentration; Lipids; Molecular Sequence Data; Phylogeny; Pigments, Biological; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Temperature; Ubiquinone

The membrane-associated antioxidant coenzyme Q10 (CoQ10) or ubiquinone-10 is frequently measured in serum or plasma. However, little is known about the total contents or redox status of CoQ10 in blood cells.. We have developed a method for determination of CoQ10 in erythrocytes. Total CoQ10 in erythrocytes was compared to the amounts of ubiquinone-10 and ubihydroquinone-10 in plasma using high-pressure liquid chromatography (HPLC) with electrochemical detection and internal standardisation (ubiquinone-9, ubihydroquinone-9).. Investigations in 10 healthy probands showed that oral intake of CoQ10 (3 mg/kg/day) led to a short-term (after 5 h, 1.57+/-0.55 pmol/microl plasma) and long-term (after 14 days, 4.00+/-1.88 pmol/microl plasma, p<0.05 vs. -1 h, 1.11+/-0.24 pmol/microl plasma) increase in plasma concentrations while decreasing the redox status of CoQ10 (after 14 days, 5.37+/-1.31% in plasma, p<0.05 vs. -1 h, 6.74+/-0.86% in plasma). However, in these healthy probands, CoQ10 content in red blood cells remained unchanged despite excessive supplementation. In addition, plasma and erythrocyte concentrations of CoQ10 were measured in five patients suffering from sickle cell anemia, a genetic anemia characterised by an overall accelerated production of reactive oxygen species. While these patients showed normal or decreased plasma levels of CoQ10 with a shifting of the redox state in favour of the oxidised part (10.8-27.2% in plasma), the erythrocyte concentrations of CoQ10 were dramatically elevated (280-1,093 pmol/10(9) ERY vs. 22.20+/-6.17 pmol/10(9) ERY).. We conclude that normal red blood cells may regulate their CoQ10 content independently from environmental supplementation, but dramatic changes may be expected under pathological conditions.

Topics: Administration, Oral; Adult; Analysis of Variance; Anemia, Sickle Cell; Antioxidants; Cholesterol; Chromatography, High Pressure Liquid; Coenzymes; Dietary Supplements; Erythrocytes; Female; Humans; Male; Time Factors; Ubiquinone

Topics: Aging; Animals; Caenorhabditis elegans; Caenorhabditis elegans Proteins; Cell Nucleus; Diet; Electron Transport; Energy Metabolism; Escherichia coli; Fermentation; Helminth Proteins; Insulin; Larva; Longevity; Mitochondria; Mutation; Reactive Oxygen Species; Receptor, Insulin; Receptors, Cytoplasmic and Nuclear; Signal Transduction; Ubiquinone

The isoprenylated benzoquinone coenzyme Q is a redox-active lipid essential for electron transport in aerobic respiration. Here, we show that withdrawal of coenzyme Q (Q) from the diet of wild-type nematodes extends adult life-span by approximately 60%. The longevity of clk-1, daf-2, daf-12, and daf-16 mutants is also extended by a Q-less diet. These results establish the importance of Q in life-span determination. The findings suggest that Q and the daf-2 pathway intersect at the mitochondria and imply that a concerted production coupled with enhanced scavenging of reactive oxygen species contributes to the substantial life-span extension.

Topics: Aging; Animals; Caenorhabditis elegans; Caenorhabditis elegans Proteins; Diet; Escherichia coli; Fermentation; Forkhead Transcription Factors; Genes, Helminth; Helminth Proteins; Larva; Longevity; Mitochondria; Models, Biological; Mutation; Oxidation-Reduction; Oxygen Consumption; Phenotype; Reactive Oxygen Species; Receptor, Insulin; Receptors, Cytoplasmic and Nuclear; Signal Transduction; Transcription Factors; Ubiquinone

Rat liver mitochondria, in different steps of the maturation process, were resolved by differential centrifugation at 1000 g (M1), 3000 g (M3), and 10,000 g (M10), and their characteristics determining susceptibility to stress conditions were investigated. Some parameters did not show gradual changes in the transition from M10 to M1 fraction because of the contamination of the M10 fraction by microsomes and damaged mitochondria with relatively high lipid content. The highest and lowest rates of O2 consumption and H2O2 production were exhibited by M1 and M10 fractions, respectively. Vitamin E and coenzyme Q levels were significantly higher in M10 than in M1 fraction, whereas whole antioxidant capacity was not significantly different. The degree of oxidative damage to lipids and proteins was higher in M1 and not significantly different in M3 and M10 fractions. The order of susceptibility to both oxidative challenge and Ca2+-induced swelling was M1 > M3 > M10. It seems that the Ca2+-induced swelling is due to permeabilization of oxidatively altered inner membrane and leads to discard mitochondria with high ROS production. If, as previous reports suggest, mitochondrial damage is initiating stimulus to mitochondrial biogenesis, the susceptibility of the M1 mitochondria to stressful conditions could be important to regulate cellular ROS production. In fact, it should favor the substitution of the oldest ROS-overproducing mitochondria with neoformed mitochondria endowed with a smaller capacity to produce free radicals.

Topics: Animals; Antioxidants; Calcium; Coenzymes; Hydrogen Peroxide; In Vitro Techniques; Male; Membrane Potentials; Mitochondria, Liver; Mitochondrial Swelling; Oxidative Stress; Oxygen Consumption; Rats; Rats, Wistar; Reactive Oxygen Species; Ubiquinone; Vitamin E

Coenzyme Q distribution, as well as respiratory chain features, in rat brain mitochondria depend on mitochondrial subpopulation, brain region and age. Heavy mitochondria (HM) usually display the lowest content of respiratory components and the lowest enzymatic activities and it has been suggested that they represent the oldest mitochondrial population. In this study, we confirmed that HM are considerably compromised in their structure. In fact, HM showed to have the highest hydroperoxide content and the most consistent modifications in their fatty acid pattern with wide loss of fatty acids (or part of them) in the phospholipid moiety. Such situation could explain the typical impairment of HM and could support the hypothesis that they represent an old mitochondrial population.

Topics: Age Factors; Aging; Animals; Antioxidants; Brain; Coenzymes; Fatty Acids; Female; Lipid Peroxidation; Lipid Peroxides; Mitochondria; Rats; Rats, Sprague-Dawley; Ubiquinone; Vitamin E

Great attention has been devoted both to ageing phenomena at the mitochondrial level and to the antioxidant status of membrane structures. These kinds of investigations are difficult to perform in the brain because of its heterogeneity. It is known that synaptic heavy mitochondria (HM) may represent an aged mitochondrial population characterized by a partial impairment of their typical mitochondrial function. We arranged a novel system requiring no extraction procedure, very limited handling of the samples and their direct injection into the HPLC apparatus, to carry out, for the first time, a systematic and concomitant determination of vitamin E, Coenzyme Q9 (CoQ9) and Coenzyme Q10 (CoQ10) contents in rat brain mitochondria. The trends found for CoQ9 and CoQ10 levels in synaptic and non-synaptic occipital cerebral cortex mitochondria during rat ageing are consistent with previous data. Hydroperoxides (HP) differed with age and it was confirmed that in the HM fraction the summation of contributions results in an oxidatively jeopardized subpopulation. We found that vitamin E seems to increase with age, at least in non-synaptic free (FM) and synaptic light (LM) mitochondria, while it was inclined to remain substantially constant in HM.

Topics: Age Factors; Aging; Animals; Antioxidants; Chromatography, High Pressure Liquid; Coenzymes; Equipment Design; Lipid Peroxidation; Lipid Peroxides; Male; Mitochondria; Occipital Lobe; Rats; Rats, Sprague-Dawley; Ubiquinone; Vitamin E

Because diabetes mellitus is associated with impairment of testicular function, ultimately leading to reduced fertility, this study was conducted to evaluate the existence of a cause-effect relationship between increased oxidative stress in diabetes and reduced mitochondrial antioxidant capacity. The susceptibility to oxidative stress and antioxidant capacity (in terms of glutathione, coenzyme Q, and vitamin E content) of testis mitochondrial preparations isolated from Goto-Kakizaki (GK) non-insulin-dependent diabetic rats and from Wistar control rats, 1 yr of age, was evaluated. It was found that GK mitochondrial preparations showed a lower susceptibility to lipid peroxidation induced by ADP/Fe(2+), as evaluated by oxygen consumption and reactive oxygen species generation. The decreased susceptibility to oxidative stress in diabetic rats was associated with an increase in mitochondrial glutathione and coenzyme Q9 contents, whereas vitamin E was not changed. These results demonstrate a higher antioxidant capacity in diabetic GK rats. We suggest this is an adaptive response of testis mitochondria to the increased oxidative damage in diabetes mellitus.

Topics: Animals; Antioxidants; Coenzymes; Diabetes Mellitus, Type 2; Disease Models, Animal; Glutathione; Glutathione Disulfide; Male; Mitochondria; Oxidative Stress; Oxygen Consumption; Rats; Rats, Inbred Strains; Rats, Wistar; Reactive Oxygen Species; Testis; Ubiquinone; Vitamin E

The characteristic pigmentation of ripe tomato fruit is due to the deposition of carotenoid pigments. In tomato, numerous colour mutants exist. The Cnr tomato mutant has a colourless, non-ripening phenotype. In this work, carotenoid formation in the Cnr mutant has been studied at the biochemical level. The carotenoid composition of Ailsa Craig (AC) and Cnr leaves was qualitatively and quantitatively similar. However, Cnr fruits had low levels of total carotenoids and lacked detectable levels of phytoene and lycopene. The presence of normal tocopherols and ubiquinone-9 levels in the ripe Cnr fruits suggested that other biosynthetically related isoprenoids were unaffected by the alterations to carotenoid biosynthesis. In vitro assays confirmed the virtual absence of phytoene synthesis in the ripe Cnr fruit. Extracts from ripe fruit of the Cnr mutant also revealed a reduced ability to synthesise the carotenoid precursor geranylgeranyl diphosphate (GGPP). These results suggest that besides affecting the first committed step in carotenoid biosynthesis (phytoene synthase) the Cnr mutation also affects the formation of the isoprenoid precursor (GGPP).

Topics: Carotenoids; Color; Lycopene; Mutation; Pigments, Biological; Plant Leaves; Polyisoprenyl Phosphates; Solanum lycopersicum; Tocopherols; Ubiquinone

The presence of the oxidized and reduced forms of ubiquinones Q(9) and Q(10) was determined in commercial extra virgin olive and seed oils, where the amounts of alpha- and gamma-tocopherols and beta-carotene were also quantitated. Very high concentrations of ubiquinones were found in soybean and corn oils. Furthermore, the total antioxidant capability of each oil was evaluated by measuring total radical-trapping antioxidant parameters (TRAP) in tert-butyl alcohol and using egg lecithin as the oxidizable substrate. These values decreased in the order sunflower > corn > peanut > olive; the highest TRAP, which was found in sunflower oil, was related to the very high amount of alpha-tocopherol. Olive oil, because of the low content of alpha-tocopherol, exhibited a TRAP value approximately one-third that of sunflower oil. TRAP values of corn and soybean oils, in which low amounts of alpha-tocopherol but very high contents of gamma-tocopherol and reduced ubiquinones were present, were intermediate. gamma-Tocopherol exhibited a poor ability of trapping peroxyl radicals in tert-butyl alcohol. This behavior was probably due to the effects of the solvent on the rate of hydrogen abstraction from this phenol.

Topics: alpha-Tocopherol; Antioxidants; Free Radicals; gamma-Tocopherol; Olive Oil; Oxidation-Reduction; Peroxides; Phenols; Plant Oils; Species Specificity; Ubiquinone

In-line connected electrochemical (EC) and diode array (DAD) detectors were compared in the reversed-phase high-performance liquid chromatographic (RP-HPLC) analysis of coenzymes Q(9) and Q(10) in some food materials (beef steak, beef heart, Baltic herring fillet, and rye flour). Coenzymes Q(9) and Q(10) were extracted from the samples using a 5:1 n-hexane-ethanol mixture. Coefficient of variation (CV%) of quadruplicate or quintuplicate determined samples for coenzymes Q(9) and Q(10) was <10 by both EC detector and DAD. Responses of the detection systems were linear in the range evaluated, 10-200 ng/injection, and had correlation coefficients exceeding 0.999. Recoveries of added coenzymes Q(9) and Q(10) varied 73-105% for DAD and 74-103% for EC detector, respectively. Detection limits for coenzymes Q(9) and Q(10) using the DAD system were 4 and 6 ng/injection, respectively, and 0.2 and 0.3 ng/injection by EC detection. Results derived from the two detection systems were generally similar. However, although EC detector was 20-fold more sensitive, the selectivity was, in some cases, poorer than that of DAD.

Topics: Animals; Cattle; Chromatography, High Pressure Liquid; Coenzymes; Electrochemistry; Fishes; Flour; Food Analysis; Meat; Secale; Swine; Ubiquinone

Consistent with the postulated role of oxidative stress in the etiology of late diabetic complications, pharmacological interventions based on biological antioxidants have been suggested. The aim of the present study was to investigate the effect of dietary supplementation with the pyridoindole antioxidant stobadine on the myocardial antioxidant status and ultrastructure of streptozotocin-diabetic rats. Diabetic male Wistar rats were fed for 32 weeks a standard diet or a diet supplemented with stobadine (0.05% w/w). Control rats received a standard diet or stobadine-supplemented diet (0.16% w/w). Plasma levels of glucose, cholesterol and triglycerides were increased significantly by diabetes. Activities of superoxide dismutase and catalase were markedly elevated in the diabetic myocardium. Myocardial levels of conjugated dienes increased after eight months of diabetes, in spite of significantly increased myocardial alpha-tocopherol and coenzyme Q9 content. The long-term treatment of diabetic animals with stobadine (i) reduced plasma cholesterol and triglyceride levels yet left the severe hyperglycemia unaffected, (ii) reduced oxidative damage of myocardial tissue as measured by conjugated dienes, (iii) reversed myocardial levels of alpha-tocopherol and coenzyme Q9 to near control values, (iv) reduced elevated activity of superoxide dismutase in the diabetic myocardium, and (v) attenuated angiopathic and atherogenic processes in the myocardium as assessed by electron microscopy examination. These results are in accordance with the postulated prooxidant role of chronic hyperglycemia and provide further evidence that development of pathological changes in diabetic myocardium is amenable to pharmacological intervention by biological antioxidants.

Topics: Animals; Anti-Arrhythmia Agents; Antioxidants; Blood Glucose; Body Weight; Carbolines; Cardiomyopathies; Catalase; Cholesterol; Diabetes Mellitus, Experimental; Drinking; Eating; Glutathione Peroxidase; Heart; Male; Myocardium; Oxidation-Reduction; Random Allocation; Rats; Rats, Wistar; Superoxide Dismutase; Triglycerides; Ubiquinone; Vitamin E

The aim of this study was to investigate if feeding with carbonyl iron would facilitate the development of preneoplastic lesions initiated by diethylnitrosamine (DEN) and promoted by CCl4-induced liver cirrhosis.. Male Wistar rats were fed a diet with 1.25%-2.5% carbonyl iron for 23 weeks and received intragastric injections of CCl4 (1.0 or 2.0 ml/kg per week) for 13 weeks, followed by one i.p. injection of DEN (200 mg/kg), after which CCl4 was administered for 8 additional weeks. Animals were killed 48 h after the first CCl4 injection to evaluate liver necrosis, 8 weeks later to evaluate fibrosis, and 9 weeks after DEN to determine formation of glutathione S-transferase 7,7 (GST-7,7) positive foci.. Treatment with iron counteracted the increased serum alanine aminotransferase levels and liver necrosis following CCl4 administration. Hepatic levels of reduced Q9 and alpha-tocopherol were elevated in rats treated with CCl4 and decreased in rats treated with iron compared to the controls. Fibrogenesis was not altered by iron treatment. Nine weeks after DEN initiation, the number and volume density of GST-7,7-positive foci in rats treated with CCl4 were significantly increased as compared with controls, but co-treatment with iron inhibited this increase. Apoptotic index was increased in iron-loaded livers, and labelling index (the fraction of S-phase hepatocytes) was decreased by co-treatment with iron in livers exposed to CCl4.. Carbonyl iron depleted hepatic levels of antioxidants, it decreased CCl4-induced necrosis and cell proliferation, it enhanced apoptosis and did not facilitate fibrogenesis. These effects together may explain the suppression of CCl4-induced promotion after DEN initiation exerted by carbonyl iron in the present study.

Topics: Animals; Antioxidants; Apoptosis; Body Weight; Carbon Tetrachloride; Carcinogens; Cell Division; Diet; Diethylnitrosamine; Iron; Kupffer Cells; Liver; Liver Neoplasms, Experimental; Male; Necrosis; Organ Size; Rats; Rats, Wistar; Ubiquinone; Vitamin E

We have studied the effects of dietary depletion of vitamin E and selenium on endogenous ubiquinone-dependent antioxidant system. Deficiency induced an increase in both coenzyme Q9 and Q10 in liver tissue, reaching a maximum between 4 and 7 weeks of deficient diet consumption. Cytochrome b5 reductase polypeptide was also enriched in membranes after 5 weeks of deficient diet consumption. Substantial DT-diaphorase activity was found in deficient, but not in control plasma membranes. Deficient membranes were very sensitive to lipid peroxidation, although a great protection was observed after incubation with NAD(P)H. Our results show that liver cells can boost endogenous ubiquinone-dependent protective mechanisms in response to deficiency in vitamin E and selenium.

Topics: Animals; Cell Membrane; Coenzymes; Cytochrome Reductases; Cytochrome-B(5) Reductase; Dihydrolipoamide Dehydrogenase; Electron Transport; Lipid Peroxidation; Liver; Male; NAD; NADP; Rats; Rats, Long-Evans; Selenium; Time Factors; Ubiquinone; Vitamin E; Vitamin E Deficiency

The main objective of this study was to determine the nature of the relationship between aging and mitochondrial coenzyme Q (CoQ) content. Mitochondria in the heart, skeletal muscle, kidney and brain of the mouse varied in both the amount of total CoQ (CoQ9 + CoQ10) content as well as in the ratio of the CoQ9 to CoQ10. CoQ content declined with age only in the skeletal muscle. Caloric restriction (CR) resulted in an increase in the amount of CoQ9 in skeletal muscle mitochondria. This effect was partially reversible upon termination of the caloric restriction regimen. Results suggest that a decrease in mitochondrial CoQ content is an integral aspect of aging in skeletal muscle.

Topics: Aging; Animals; Brain; Coenzymes; Heart; Kidney; Male; Mice; Mice, Inbred C57BL; Mitochondria; Mitochondria, Heart; Mitochondria, Muscle; Muscle Development; Muscle, Skeletal; Organ Specificity; Ubiquinone

The present study was undertaken to investigate possible effects of dietary iron during the progression step in hepatocarcinogenesis.. Two experiments were performed, in which preneoplastic foci were produced in rat liver using the Solt & Farber protocol, with diethylnitrosamine as initiator and partial hepatectomy + 2-acetylaminofluorene as promoter. Two weeks after promotion, animals were fed 1.25-2.5% dietary carbonyl iron or a control diet until sacrifice. In the first experiment, animals were killed at different time points when they developed an abdominal mass in combination with weight loss. In the second experiment, animals were sacrificed 45 weeks post-promotion. Liver tumours were counted and histologically graded. Tumour levels of ubiquinone-9 and alpha-tocopherol were determined with HPLC, and labelling and apoptotic indices calculated using immunohistochemistry. The number and area of glutathione S-transferase 7,7 (GST-7,7)-positive foci were determined.. In experiment number 1, survival and tumour differentiation were similar in iron-treated animals and controls. In the second experiment, iron-treated rats had an increased number of GST-7,7-positive foci compared to controls. Number and size of carcinomas were similar between the groups, whereas tumour differentiation was higher in rats exposed to iron. Cell proliferation, apoptosis and concentrations of alpha-tocopherol in tumours were not altered by iron. The ratio of reduced/oxidized ubiquinone-9 was decreased in tumours from iron-treated animals.. In this model, dietary iron overload resulted in an increased number of preneoplastic foci but did not enhance the progression of these into hepatocellular carcinomas. Iron decreased the ratio of reduced/oxidized ubiquinone-9 in tumours, indicating that neoplastic liver cells utilize intracellular ubiquinones as a defense mechanism against iron-induced oxidative stress.

Topics: 2-Acetylaminofluorene; Animals; Apoptosis; Carcinogens; Carcinoma, Hepatocellular; Cell Division; Chromatography, High Pressure Liquid; Diethylnitrosamine; Disease Progression; Glutathione Transferase; Hemochromatosis; Iron; Iron, Dietary; Liver; Liver Neoplasms, Experimental; Male; Oxidative Stress; Rats; Rats, Wistar; Ubiquinone; Vitamin E

We have studied the interaction of coenzyme Q with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and its metabolites, 1-methyl-4-phenyl-2,3-dihydropyridinium (MPDP(+)) and 1-methyl-4-phenylpyridinium (MPP(+)), the real neurotoxin to cause Parkinson's disease. Incubation of MPTP or MPDP(+) with rat brain synaptosomes induced complete reduction of endogenous ubiquinone-9 and ubiquinone-10 to corresponding ubiquinols. The reduction occurred in a time- and MPTP/MPDP(+) concentration-dependent manner. The reduction of ubiquinone induced by MPDP(+) went much faster than that by MPTP. MPTP did not reduce liposome-trapped ubiquinone-10, but MPDP(+) did. The real toxin MPP(+) did not reduce ubiquinone in either of the systems. The reduction by MPTP but not MPDP(+) was completely prevented by pargyline, a type B monoamine oxidase (MAO-B) inhibitor, in the synaptosomes. The results indicate that involvement of MAO-B is critical for the reduction of ubiquinone by MPTP but that MPDP(+) is a reductant of ubiquinone per se. It is suggested that ubiquinone could be an electron acceptor from MPDP(+) and promote the conversion from MPDP(+) to MPP(+) in vivo, thus accelerating the neurotoxicity of MPTP.

Topics: 1-Methyl-4-phenylpyridinium; Animals; Biotransformation; Liposomes; Male; Monoamine Oxidase; Neurotoxins; Oxidation-Reduction; Pyridinium Compounds; Rats; Rats, Wistar; Synaptosomes; Ubiquinone

A high-performance liquid chromatographic (HPLC) method is presented for the simultaneous detection of ubiquinone-9 and 10 in rat tissues such as blood, myocardium, and muscle. After liquid-liquid extraction, the ubiquinones are subsequently analyzed by HPLC with ultraviolet (UV) detection at their maximum absorbance (275 nm). Reference calibration curves in ethanol are used to determine tissular levels of ubiquinones. Because a treatment with HMG-CoA reductase inhibitors is expected to decrease the ubiquinone levels, reference calibration curves are performed to ensure that the ratios (ubiquinone/internal standard) observed in such an experiment could be evaluated directly on a calibration curve. The assay is sensitive (0.0625 microgram/mL), reproducible (4% coefficient of variation for ubiquinone-9 and 6% for ubiquinone-10), and linear up to 20 micrograms/mL (or 100 mg of tissue) for ubiquinone-9 and up to 10 micrograms/mL (or 100 mg of tissue) for ubiquinone-10. The ubiquinone levels in control tissues or blood are within the ranges of those previously reported.

Topics: Animals; Calibration; Chromatography, High Pressure Liquid; Circadian Rhythm; Ethanol; Linear Models; Male; Muscles; Myocardium; Osmolar Concentration; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Sensitivity and Specificity; Spectrophotometry, Ultraviolet; Ubiquinone

S-Allylmercaptocysteine (SAMC), one of the water-soluble organosulfur compounds in ethanol extracts of garlic (Allium sativum L.), has been shown to protect mice against acetaminophen (APAP)-induced liver injury. In this study, we examined the mechanisms underlying this hepatoprotection. SAMC (100 mg/kg, p.o.) given 2 and 24 hr before APAP administration (500 mg/kg, p.o.) suppressed the plasma alanine aminotransferase activity increases 3 to 12 hr after APAP administration significantly. The hepatic reduced glutathione levels of vehicle-pretreated mice decreased 1 to 6 hr after APAP administration, but SAMC pretreatment suppressed the reductions 1 to 6 hr after APAP administration significantly. These inhibitory effects of SAMC were dose-dependent (50-200 mg/kg) 6 hr after APAP administration. As SAMC pretreatment (50-200 mg/kg) suppressed hepatic cytochrome P450 2E1-dependent N-nitrosodimethylamine demethylase activity significantly in a dose-dependent manner, we suggest that one of its protective mechanisms is inhibition of cytochrome P450 2E1 activity. SAMC pretreatment also suppressed the increase in hepatic lipid peroxidation and the decrease in hepatic reduced coenzyme Q9 (CoQ9H2) levels 6 hr after APAP administration. The hepatic CoQ9H2 content of the SAMC pretreatment group was maintained at the normal level. Therefore, we suggest that another hepatoprotective mechanism of SAMC may be attributable to its antioxidant activity.

Topics: Acetaminophen; Alanine Transaminase; Animals; Chemical and Drug Induced Liver Injury; Coenzymes; Cysteine; Cytochrome P-450 CYP2E1; Glucuronosyltransferase; Glutathione; Lipid Peroxidation; Liver; Liver Diseases; Male; Mice; Proteins; Sulfhydryl Compounds; Sulfotransferases; Ubiquinone; Vitamin E

We examined effects of pravastatin on age-related changes in mitochondrial function in rats. Decline in the activity of complex I of the mitochondrial electron transport chain was observed in diaphragm and psoai major in rats aged 35 and 55 weeks, and that of complex IV in rats aged 55 weeks. Pravastatin accelerated significantly age-related decline in the activity of complex I of diaphragm mitochondria, though pravastatin did not show significant effect on normally observed age-associated decline in the activities of complex IV of psoai major and diaphragm mitochondria. Aging effect on mitochondrial respiratory function was not observed on heart muscle and liver in rats up to 55 weeks old, and pravastatin did not effect significantly heart and liver mitochondrial respiratory function. From these results, careful clinical examination on respiratory muscle function should be necessary in patients treated with pravastatin particularly in elderly patients.

Topics: Aging; Animals; Cell Respiration; Diaphragm; Electron Transport; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Male; Mitochondria, Heart; Mitochondria, Liver; Mitochondria, Muscle; Multienzyme Complexes; NAD(P)H Dehydrogenase (Quinone); Oxidoreductases; Pravastatin; Rats; Rats, Wistar; Specific Pathogen-Free Organisms; Ubiquinone

To confirm whether or not cytosolic NADPH-UQ reductase is involved in the recycling of cellular ubiquinol (UQH2) consumed during lipid peroxidation, the effect of a UQ-10 supplement on the NADPH-UQ reductase and cellular defense against oxidative damage in rat livers was investigated. Supplements of UQ-10 for 14 days enhanced the levels of UQH2-10 and NADPH-UQ reductase in rat livers without any appreciable changes in other antioxidant contents and related enzyme activities. However, the injection of carbon tetrachloride (CCl4) into the rats induced lipid peroxidation and decreased the cellular UQH2-10 contents (and increased equivalent amounts of UQ-10), as well as decreasing the ascorbic acid, reduced glutathione (GSH) and alpha-tocopherol contents of the rat livers. Administration of the UQ-10 supplement prior to the CCl4 treatment spared alpha-tocopherol (but not GSH or ascorbic acid), inhibited lipid peroxidation, and thus improved CCl4-induced hepatitis. These findings support the notion that NADPH-UQ reductase in cytosol is the enzyme responsible for the regeneration of UQH2 from UQ formed by lipid peroxidation in cells.

Topics: Animals; Carbon Tetrachloride; Chemical and Drug Induced Liver Injury; Cytosol; Liver; Male; Microsomes, Liver; Mitochondria, Liver; NAD(P)H Dehydrogenase (Quinone); Oxidation-Reduction; Rats; Rats, Wistar; Specific Pathogen-Free Organisms; Ubiquinone

The presence of Coenzyme Q (CoQ) in food, its role in cellular bioenergetics and antioxidant protection and the key role played by dietary fatty acids on membrane structure support the interest for a wide research concerning the relationship between dietary fats, CoQ content and biochemical behaviour. Several models of peroxidative stress 'in vivo' have been extensively investigated in our laboratory, with particular regards to the influence of dietary fat upon mitochondrial CoQ levels. First studies showed that the unsaturation degree of dietary fat leads to different CoQ9 and CoQ10 mitochondrial contents. The highest levels were found using polyunsaturated fat. A significant CoQ9 decrease after adriamycin peroxidative induction was found when dietary fat was polyunsaturated; on the contrary, a light increase was found in the case of monounsaturated fat. Another example of oxidative stress is that produced by food frying. The results obtained were in some cases similar to those of the previous experimental design: in fact monounsaturated dietary fats increased CoQ mitochondrial contents, whereas the polyunsaturated ones decreased CoQ levels. Finally, the combined effect of physical exercise and dietary fats on tissue and plasma CoQ levels has been studied. CoQ levels did not change during aerobic performances when dietary fat was monounsaturated whereas light increases were detected in the case of polyunsaturated fats. On the contrary, in anaerobic conditions, CoQ levels clearly increased with monounsaturated fats and no alterations were found in the case of polyunsaturated ones.

Topics: Aerobiosis; Anaerobiosis; Coenzymes; Cooking; Corn Oil; Dietary Fats; Doxorubicin; Hot Temperature; Lipid Peroxidation; Malondialdehyde; Membrane Lipids; Mitochondria; Olive Oil; Oxidative Stress; Physical Exertion; Plant Oils; Sunflower Oil; Ubiquinone; Vitamin E

The individual and combined effects of ethanol and lovastatin on rats and their prevention by supplemental coenzyme Q10 (CoQ10) was studied. The ethanol and lovastatin findings are reported elsewhere. This paper focuses on the food restriction which occurred in rats fed 35% of energy as ethanol and those control rats pair-fed to the 35% of energy as ethanol group. Six groups of rats received 35% of energy as ethanol (with or without lovastatin and/or CoQ10 treatment). One group served as a 0% ethanol ad libitum control and one 0% ethanol control group was pair-fed to the 35% ethanol group. Rats receiving 35% of energy as ethanol and their pair-fed controls consumed 83% of the energy/day consumed by the ad libitum controls. This was consistent regardless of lovastatin or CoQ10 treatment. Weight gains were 84% of control. The energy reduction was consistently associated with a substantial (48%+) increase in liver CoQ9 concentrations. Reports by others of associations between food restriction and increased longevity in rodents has focused on a decrease in oxidant damage in tissues of food restricted animals. The increase in CoQ levels in the food restricted animals would result in an increase in antioxidant protection and might explain the observed increases in longevity.

Topics: Animal Feed; Animals; Antioxidants; Appetite; Electrocardiography; Ethanol; Exercise Test; Food Deprivation; Food, Formulated; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Lipid Metabolism; Liver; Lovastatin; Myocardium; Rats; Ubiquinone

Six experimental groups of young (7-month-old) and aged (24-32-month-old) rats, underwent different dietary manipulations (i.e. dietary restriction and/or a vitamin E-depleted diet), and their liver mitochondria were assayed for several antioxidants and peroxidation markers. Glutathione levels were affected both by age and dietary treatment. Coenzyme Q9 and C0Q10 showed the highest levels in the oldest rats where ageing, as well as other oxidative stresses, could induce ubiquinone biosynthesis.

Topics: Aging; Animals; Antioxidants; Coenzymes; Food Deprivation; Glutathione; Hydrogen Peroxide; Lipid Peroxidation; Longevity; Mitochondria, Liver; Oxidative Stress; Rats; Ubiquinone; Vitamin E Deficiency

The levels of coenzyme Q were determined in blood plasma and regenerating liver mitochondria of hepatectomized rats, using as controls either sham-operated or non-operated animals. Mitochondrial CoQ9 content increased in sham-operated rats, whereas it was significantly lower in hepatectomized with respect to non-operated animals. Plasma CoQ9 levels decreased dramatically in hepatectomized animals, but increased strongly in sham-operated in comparison with non-operated rats. The data suggest the possibility of a rate-limiting step in CoQ biosynthesis in hepatectomized animals.

Topics: Animals; Hepatectomy; Lipoproteins; Liver Regeneration; Male; Mitochondria, Liver; Postoperative Period; Rats; Rats, Wistar; Ubiquinone

In order to examine whether iron and alcohol act synergistically during tumor initiation in vivo, we investigated the effects of dietary iron overload and a liquid ethanol-containing diet on the initiation phase of the Solt & Farber model of chemical hepatocarcinogenesis.. Following dietary supplementation with carbonyl iron for 8 weeks and ethanol pair-feeding according to Lieber deCarli for 5 weeks, animals were subjected to partial hepatectomy in order to induce regenerative cell proliferation and thereby "fix" putative DNA lesions. Levels of malondialdehyde, reduced and oxidized ubiquinone-9, alpha-tocopherol and 8-oxo-2'-deoxyguanosine were analyzed in liver tissue removed at the time of partial hepatectomy, and blood was collected for determination of alanine amino-transferase activities. Following a 2-week recovery period, promotion was achieved with 0.02% dietary 2-acetylaminofluorene and carbon tetrachloride. Two weeks after the completion of promotion, animals were sacrificed and the number of preneoplastic, glutathione S-transferase 7,7-positive lesions counted. Animals initiated with diethylnitrosamine served as a positive control group.. Serum aminotransferase activities were significantly increased, and hepatic contents of ubiquinol-9 (reduced ubiquinone-9) were significantly decreased in animals exposed to the combination of iron and ethanol in comparison to the other groups. Livers from iron-treated animals had decreased levels of alpha-tocopherol and increased contents of malondialdehyde, whereas treatment with ethanol did not further enhance these alterations. Levels of 8-oxo-2'-deoxyguanosine were not significantly different in animals treated with iron, ethanol or iron + ethanol as compared with controls. The number of preneoplastic foci at the time of sacrifice was not increased in livers exposed to iron and/or ethanol as compared with those from control animals. As expected, the number of foci was significantly increased in positive controls which were initiated with diethylnitrosamine.. Iron potentiated the cytotoxic effects of ethanol, resulting in increased serum aminotransferase activities and decreased hepatic contents of ubiquinol. However, the combination of iron and ethanol did not exert genotoxic effects detectable as enhanced hepatic levels of 8-oxo-2'-deoxyguanosine, or increased formation of preneoplastic, glutathione S-transferase 7,7-positive lesions in the Solt & Farber model of chemical hepatocarcinogenesis.

Topics: Alanine Transaminase; Animals; Antioxidants; Body Weight; Disease Models, Animal; Ethanol; Iron Overload; Liver; Liver Neoplasms, Experimental; Male; Malondialdehyde; Rats; Rats, Sprague-Dawley; Rats, Wistar; Ubiquinone; Vitamin E

Different organisms produce different species of isoprenoid quinones, each with its own distinctive length. These differences in length are commonly exploited in microbial classification. The side chain length of quinone is determined by the nature of the polyprenyl diphosphate synthase that catalyzes the reaction. To determine if the side chain length of ubiquinone (UQ) has any distinct role to play in the metabolism of the cells in which it is found, we cloned the solanesyl diphosphate synthase gene (sdsA) from Rhodobacter capsulatus SB1003 and expressed it in Escherichia coli and Saccharomyces cerevisiae. Sequence analysis revealed that the sdsA gene encodes a 325-amino-acid protein which has similarity (27 to 40%) with other prenyl diphosphate synthases. Expression of the sdsA gene complemented a defect in the octaprenyl diphosphate synthase gene of E. coli and the nonrespiratory phenotype resulting from a defect in the hexaprenyl diphosphate synthase gene of S. cerevisiae. Both E. coli and S. cerevisiae expressing the sdsA gene mainly produced solanesyl diphosphate, which resulted in the synthesis of UQ-9 without any noticeable effect on the growth of the cells. Thus, it appears that UQ-9 can replace the function of UQ-8 in E. coli and UQ-6 in S. cerevisiae. Taken together with previous results, the results described here imply that the side chain length of UQ is not a critical factor for the survival of microorganisms.

Topics: Alkyl and Aryl Transferases; Amino Acid Sequence; Cloning, Molecular; Dimethylallyltranstransferase; Escherichia coli; Genetic Complementation Test; Molecular Sequence Data; Polyisoprenyl Phosphates; Polymerase Chain Reaction; Rhodobacter capsulatus; Saccharomyces cerevisiae; Sequence Alignment; Transferases; Ubiquinone

1. The effect of ubiquinone (CoQ(9)) on ascites in broiler chickens was investigated. 2. The commercial broilers were divided into 2 groups of 100 birds each; CoQ(9) treated group and non-treated group. 3. The chickens were grown in a positive-pressured house with double high efficiency particulate air filtered intakes and exhaust, and thus were strictly isolated from infectious agents. 4. The chickens (15 to 21 d old) were exposed to cold stress in order to induce ascites. 5. The number of birds with ascites in the CoQ(9)-treated group was significantly lower than in the non-treated group. 6. Survival and production rates were better in the CoQ(9)-treated group than in the non-treated group.

Topics: Animal Feed; Animals; Antioxidants; Ascites; Chickens; Cold Temperature; Food, Fortified; Hematocrit; Male; Poultry Diseases; Stress, Physiological; Ubiquinone

Coenzyme Q content was monitored in blood plasma and regenerating liver mitochondria of hepatectomized rats, using as controls either sham-operated or non-operated animals. Mitochondrial CoQ9 content increased in sham-operated rats, whilst it was significantly lower in hepatectomized in comparison with non-operated animals at all considered times. On the other hand plasma CoQ9 levels dramatically decreased in hepatectomized animals, while strongly increased in sham-operated in comparison with non-operated rats. The quinone decrease in hepatectomized animals is likely to be due to the attainment of a rate-limiting step in CoQ biosynthesis.

Topics: Animals; Coenzymes; Hepatectomy; Liver Regeneration; Male; Mitochondria, Liver; Rats; Rats, Wistar; Ubiquinone

The possibility of reconstituting a functionally competent endogenous plastoquinone pool in photosystem II (PS II) membrane fragments, inside-out-vesicles (ISO-vesicles), and PS II core complexes was analyzed by measuring (i) the characteristic period four oscillation of the oxygen yield due to excitation of dark-adapted samples with a train of short flashes and (ii) laser flash-induced transients of the relative quantum yield of chlorophyll fluorescence. The data obtained revealed that (a) an endogenous pool capacity comparable to that of intact thylakoids can be restored in PS II membrane fragments and ISO-vesicles by a sonication treatment using native plastoquinone-9, (b) a more pronounced oxygen oscillation pattern arises in PS II core complexes after application of the same reconstitution procedure, (c) the extent of the endogenous pool restoration at a ratio of 15 quinone molecules per PS II in the reconstitution assay strongly depends on the nature of the quinone molecule [maximum effects can be only achieved with PQ-9, while at the same concentration ubiquinone-45 (UQ-9) is almost inefficient], and (d) a sonication step is required for stable insertion of PQ-9 into PS II preparations. Measurements of the reconstruction degree as a function of the structure of different quinones with selected properties lead to the conclusion that specific binding domains exist in PS II in addition to the QB site. These domains exhibit a surprisingly high specificity for the type of quinone that can be bound. On the basis of a comparison of the results obtained, the structure of the quinone head group seems to be more important than the large hydrophobic side chain and/or the general lipophilicity of the compound.

Topics: Fluorescence; Intracellular Membranes; Kinetics; Light-Harvesting Protein Complexes; Oxygen; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Plastoquinone; Spinacia oleracea; Ubiquinone

Physical exercise increases metabolic rate, and induces both adaptational biogenesis of mitochondria in skeletal muscle and an increase in antioxidant capacity. The onset of experimental anorexia and cachexia can be delayed by voluntary exercise. As skeletal muscle is the main target for cancer cachexia, we determined the levels of coenzymes Q9 and Q10 in skeletal muscle from tumour-bearing exercising rats, and compared them to those of sedentary tumour-bearers and controls. Both tumour-bearing groups had increased levels of coenzymes Q9 and Q10 in the anterior tibial muscle (P < 0.05 for exercised animals). In the soleus muscle, only the tumour-bearing exercising animals demonstrated an increase in the levels of both coenzymes (P < 0.05). In cardiac muscle, the presence of tumour and exercise reduced the levels of coenzymes below that of sedentary controls. Exercise counteracted the anaemia in the tumour-bearing host (P < 0.05). In conclusion, the increase in antioxidant capacity in skeletal muscle indicates a defence mechanism in the tumour-bearing hosts which is augmented by physical exercise.

Topics: Animals; Cachexia; Coenzymes; Energy Metabolism; Female; Muscle, Skeletal; Myocardium; Neoplasms, Experimental; Physical Conditioning, Animal; Rats; Rats, Inbred WF; Ubiquinone

The dynamics of T and B cell immunity in spleens from rat exposed to whole-body chronic irradiation with dose rates of 12.9 cGy/day (range 1-10 Gy) and 3.0 cGy/day (range 0.57-2.04 Gy) were investigated. gamma-irradiation with a dose-rate of 12.9 cGy/day was shown to produce a wave-like suppression of the T lymphocyte mitogenic response. Irradiation with a dose-rate of 3.0 cGy/day caused a decrease in immune response of T lymphocytes 48 days after onset of exposure (total dose 1.4 Gy). It was also shown that chronic irradiation with a dose-rate of 3.0 cGy/day produced significant changes in the DNA of T lymphocytes. Our results show that the radiation-induced suppression of immune functions and damage to DNA structure were partially eliminated when animals were fed a daily diet supplemented with a natural antioxidant, ubiquinone Q-9. The inhibiting effect of chronic irradiation was more pronounced in B lymphocytes because of their higher radiosensitivity.

Topics: Animals; Antioxidants; B-Lymphocytes; Diet; DNA; Dose-Response Relationship, Radiation; Flow Cytometry; Immune System; Male; Radiation Dosage; Rats; Rats, Wistar; Spleen; T-Lymphocytes; Ubiquinone

Active oxygen species are reported to cause organ damage. This study was therefore designed to determine the behaviour of antioxidants and free radical scavengers so as to reveal changes in animals in the hyper- and hypothyroid state. Levels of antioxidant factors (i.e. coenzyme Q (CoQ)10, CoQ9 and vitamin E) and free radical scavengers (catalase, glutathione peroxidase (GSH-PX) and superoxide dismutase (SOD)) were measured in the heart muscles of rats rendered hyper- or hypothyroid by 4 weeks of thyroxine (T4) or methimazol treatment. Serum levels of CoQ9 and total SOD were also measured. A significant reduction in CoQ9 levels was observed in the heart muscles of both hyper- and hypothyroid rats when compared with control hearts. There was no difference in serum CoQ9 levels in thyroid dysfunction when compared with control animals. Levels of vitamin E in the heart muscles of hyperthyroid rats were significantly increased, and there was no reduction in vitamin E levels in hypothyroid rats when compared with control hearts. GSH-PX levels in the heart muscle were reduced in hyperthyroid rats and increased in hypothyroid rats when compared with control hearts. However, there were no differences in catalase levels in heart muscle between hyper- and hypothyroid rats. The concentration of SOD in heart muscle was increased in hyperthyroid rats and was not decreased in hypothyroid rats compared with control rats, suggesting the induction of SOD by excessive production of O2-. These data suggest that the changes in these scavengers have some role in cardiac dysfunction in the hyper- and hypothyroid state in the rat.

Topics: Animals; Catalase; Cholesterol; Coenzymes; Free Radical Scavengers; Glutathione Peroxidase; Male; Methimazole; Myocardium; Rats; Rats, Wistar; Superoxide Dismutase; Thyroid Hormones; Thyroxine; Triglycerides; Ubiquinone; Vitamin E

Mitochondria-rich fractions isolated from livers of rats fed diets differing in their vitamin E (E) and/or selenium (Se) contents were subjected to NADPH/ADP/Fe(3+)- dependent assays of lipid peroxidation. Addition of GSH resulted in an inhibition, or lag period, of lipid peroxidation in mitochondria from rats supplemented with E. This effect was independent of the Se status of the rats. Addition of GSH + GSSG did not potentiate the lag period over that observed with GSH alone. Significant changes in mitochondrial alpha-TH during lipid peroxidation, either in the presence or absence of GSH, were not observed. Total protein thiol (PrSH) content of native mitochondria was lower in rats fed a diet deficient in both E and Se, compared to the other dietary groups. Addition of GSH or GSH + GSSG maintained mitochondrial PrSH at higher levels during lipid peroxidation than in control assays without added GSH/GSSG. Addition of GSSG alone decreased PrSH in mitochondria prepared from all rats regardless of their E or Se status. Reduced ubiquinone-9 (U-9) and the % of total U-9 and U-10 in the reduced form were significantly decreased in liver tissue from rats fed the diet deficient in both E and Se.

Topics: Animals; Diet; Glutathione; Glutathione Peroxidase; In Vitro Techniques; Lipid Peroxidation; Male; Mitochondria, Liver; Oxidation-Reduction; Oxidative Stress; Rats; Selenium; Sulfhydryl Compounds; Ubiquinone; Vitamin E

The effect of long-term (18 months) selenium deficiency on the levels of liver coenzyme Q was studied in the rat. Levels of coenzyme Q9 and coenzyme Q10 in the liver of selenium-deficient rats were 40 and 67% of the levels in selenium-adequate animals, respectively. The results are similar to the findings using a shorter feeding period.

Topics: Animals; Coenzymes; Glutathione Transferase; Iodide Peroxidase; Liver; Male; Rats; Rats, Wistar; Selenium; Thyroid Hormones; Ubiquinone

Topics: 8-Hydroxy-2'-Deoxyguanosine; Animals; Antioxidants; Ascorbic Acid; Body Composition; Deoxyguanosine; DNA Damage; Male; Microbodies; Organ Specificity; Oxidation-Reduction; Oxidative Stress; Pyrimidines; Rats; Rats, Inbred F344; Ubiquinone; Uric Acid; Vitamin E

Alcohol metabolism may result in oxidant stress and free radical injury through a variety of mechanisms. Lovastatin may also produce oxidant stress by reducing levels of an endogenous antioxidant, coenzyme Q (CoQ). The separate and combined effects of ethanol, 20 EN% in a total liquid diet, and lovastatin, 67 mg/kg diet, on alpha-tocopherol, retinol palmitate, CoQ9 and thiobarbituric acid reactive (TBAR) material in liver from rats were determined. The effect of exogenous CoQ10 on these treatment groups was also determined. Food consumption, weight gain, liver lipid and TBAR material were similar between treatment groups. Compared to control animals, ethanol reduced retinol palmitate significantly, from 143 to 90 micrograms/g wet weight. Lovastatin had no effect on retinal palmitate nor did it act additively with ethanol. Ethanol decreased liver alpha-tocopherol from 28 to 12 micrograms/g wet weight and lovastatin diminished it to 12 micrograms; no additive effect was evident. Ethanol had no effect, but lovastatin decreased CoQ9 from 83 to 55 micrograms/g wet weight. Supplementation with CoQ10 did not modulate the effect of ethanol on retinal palmitate, but it did reverse the effect of lovastatin on CoQ9. Supplementary CoQ10 did not alter control levels of alpha-tocopherol, but it appeared to reverse most of the decrease in alpha-tocopherol attributable to ethanol or lovastatin separately. It only partially reversed the effect of ethanol and lovastatin combined on alpha-tocopherol, however. As expected, lovastatin had no effect on CoQ10 levels in supplemented animals. Ethanol, either separately or in combination with lovastatin, diminished liver stores of CoQ10 by almost 40%. We conclude that 20 EN% ethanol given in a liquid diet for 5 weeks is sufficient to lower retinol palmitate and that lovastatin reduces CoQ9. Both diminish alpha-tocopherol, an effect largely overcome by CoQ10 supplementation with either drug alone, but not with the combination. Since many individuals chronically consume the levels of ethanol represented by this experiment, and since a certain number of those also take lovastatin, further research into the possible clinical significance of these observations is warranted.

Topics: Animals; Antioxidants; Body Weight; Coenzymes; Diet; Diterpenes; Drug Evaluation, Preclinical; Ethanol; Liver; Liver Diseases, Alcoholic; Lovastatin; Organ Size; Oxidative Stress; Rats; Rats, Sprague-Dawley; Retinyl Esters; Thiobarbituric Acid Reactive Substances; Ubiquinone; Vitamin A; Vitamin E

The system of perfusing rat livers has been used to evaluate the uptake and incorporation of liposomal CoQ10 into mitochondria. After 90 minutes of perfusion the cells are strongly enriched in CoQ10 up to levels of the same order of magnitude as CoQ9. Heavy and light mitochondrial crude subcellular fractions, low in CoQ10 in control livers, contain high amounts of the quinone after perfusion; yet the purification of these fractions on a metrizamide gradient reveals that the exogenous quinone is mainly associated with the light mitochondrial subfraction, enriched in lysosomes. An increase of the NAD-dependent glutamate-malate oxidase activity is observed in CoQ10 perfused animals. As the total levels of CoQ9 + CoQ10 in these animals are not significantly modified by the CoQ10 incorporated, the observed higher activity is not ascribable to an integration of exogenous quinone into the ubiquinone pool. An antioxidant effect of extramitochondrial CoQ10 on mitochondrial functions is suggested.

Topics: Animals; Cell Fractionation; Liposomes; Male; Metrizamide; Mitochondria, Liver; Perfusion; Phosphatidylcholines; Phospholipids; Rats; Rats, Wistar; Subcellular Fractions; Ubiquinone

In previous studies we have found that a single acute dose of ultraviolet radiation to murine skin causes a large degree of destruction of enzymic and non-enzymic antioxidants immediately after irradiation. In the present study, we wished to elucidate the recovery of antioxidants after a single dose of ultraviolet (UV) radiation. We measured antioxidants and lipid hydroperoxides (as a marker of membrane damage) in murine epidermis and the dermis at 0, 3, 12, 24, 72 and 120 h after exposure to UV radiation (25 J/cm2, UVA+UVB). Lipid hydroperoxides showed the highest values immediately after UV exposure and returned to control values within 24 h in both epidermis and dermis. The activities of catalase, glutathione peroxidase and glutathione reductase showed the lowest activities immediately after UV exposure; superoxide dismutase activities reached a minimum at 3 h postexposure. The pattern of recovery was different for each enzyme and for epidermis and dermis. The activities of superoxide dismutase and catalase decreased remarkably and recovered slowly. Superoxide dismutase in the dermis recovered full activity by 120 h and in the epidermis by 12 h. Catalase activity in both epidermis and dermis had returned to only 50% of control activity at 120 h, although the epidermis showed a temporary increase (to 93%) at 24 h. Glutathione peroxidase and glutathione reductase were slightly decreased immediately after irradiation, recovered to 100% at 3 h and then increased to 200-250% in both the epidermis and the dermis at various times; values had returned to 100% in epidermis by 120 h but remained elevated in dermis.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Antioxidants; Ascorbic Acid; Catalase; Dehydroascorbic Acid; Epidermis; Female; Glutathione; Glutathione Disulfide; Glutathione Peroxidase; Glutathione Reductase; Lipid Peroxides; Mice; Mice, Hairless; Radiation Dosage; Skin; Superoxide Dismutase; Time Factors; Ubiquinone; Ultraviolet Rays; Vitamin E

Cultured neurons from rat dorsal root ganglia and cerebral cortex were infected with Sendai virus, which gives a productive replication with lysis of most neurons, and with the RW strain of mumps virus, which undergoes defective replication causing degeneration of only 30-40% of the neurons within 5 days after initial infection. In Sendai virus-infected cells the amount of polyisoprenoid lipids was enhanced. In mumps virus-infected cultures there were transient reductions in the contents of cholesterol, dolichol, and ubiquinone-9 in the cultures, whereas the reduction in the ubiquinone-10 level was progressive, reaching 20% of its original value 21 days after infection. Treatment of mumps virus-infected cultures with ubiquinone-10 protected the neurons from degeneration, whereas no effects were observed on exposure to ubiquinone-9. Linolenic acid (18:3) and arachidonic acid (20:4), but not myristic acid (14:0) and palmitic acid (16:0), also had significant neuroprotective effects.

Topics: Animals; Cell Survival; Cells, Cultured; Cerebral Cortex; Cholesterol; Dolichols; Fatty Acids, Nonesterified; Ganglia, Spinal; Kinetics; Mumps virus; Nerve Degeneration; Neurons; Parainfluenza Virus 1, Human; Peroxides; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; tert-Butylhydroperoxide; Time Factors; Ubiquinone; Virus Replication

The purpose of the present study was to evaluate the biosynthesis of coenzyme Q9 (CoQ9) in isolated and perfused young (6 months) and aged (24 months) rat hearts, either under aerobic perfusion condition or during postischemic reperfusion. The young and aged hearts have been divided into 2 groups: Group A, aerobic perfusion for 60 min with recirculating Krebs-Henseleit solution, containing 0.8 microM p-OH-[U-14C]benzoate plus 2.5 mM mevalonlactone; Group B, severe ischemic perfusion for 30 min, followed by 60 min of reperfusion under the same experimental condition of Group A. At the end of the reperfusion the mitochondrial content of CoQ9 was lower in young than aged rat hearts (p < 0.01). In Group A the incorporation of the labeled precursor into mitochondrial CoQ9 was greater in the hearts of aged than young rats (p < 0.01); on the contrary, in Group B this incorporation was significantly reduced in aged than in young rats (p < 0.05). Thus, it is possible that, in the aged rat heart, the higher activity of CoQ9 biosynthesis is related to an elevated turnover of the coenzyme due to the aging process; moreover, this activity is partially reduced by an ischemic-reperfusion stress.

Topics: Aerobiosis; Aging; Animals; In Vitro Techniques; Male; Myocardial Ischemia; Myocardial Reperfusion; Myocardial Reperfusion Injury; Myocardium; Rats; Rats, Wistar; Ubiquinone

The influence of continuous gamma irradiation on the lipids of nuclei and chromatin of rat liver at a dose-rate of 0,129 Gy/day for 155 days (a total dose of 20 Gy) and by feeding of ubiquinone-9 has been studied. The amount of phosphatidylcholine with phosphatidylserine and phosphatidyl-ethanolamine in liver nuclei of irradiated rats was found to increase. Ubiquinone-9 had a normalizing effect. A decrease of cardiolipin was observed in the liver chromatin of irradiated rats. The amount of free fatty acids had a tendency to decrease in homogenate, nuclei and liver chromatin of irradiated rats. Ubiquinone was found to increase the amount of free fatty acids up to the control level. The amount of cholesterol in nuclei was increased after irradiation and that in chromatin tended to rise. Ubiquinone-9 significantly decreased the amount of cholesterol in nuclei and chromatin of irradiated rats.

Topics: Animals; Cell Nucleus; Cholesterol; Chromatin; Chromatography, Thin Layer; Fatty Acids, Nonesterified; In Vitro Techniques; Lipids; Liver; Male; Phospholipids; Radiation Dosage; Rats; Rats, Wistar; Ubiquinone

The half-life of ubiquinone-9 in various rat tissues was determined. Rats were injected intraperitoneally with [3H]mevalonate and the decay of radioactivity incorporated into ubiquinone-9 was followed using reverse-phase HPLC. The half-life varied between 49 h (testis) and 125 h (kidney).

Topics: Animals; Chromatography, High Pressure Liquid; Half-Life; Male; Rats; Rats, Sprague-Dawley; Ubiquinone

A possible difference in antioxidant activity between reduced coenzyme Q9 (CoQ9H2) and reduced coenzyme Q10 (CoQ10H2) in animal cells was studied by incubation of hepatocytes with a hydrophilic radical initiator, 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH). Two kinds of hepatocytes differing in their content of CoQ homologs were used: rat, total (oxidized plus reduced) CoQ9: total CoQ10 6:1, guinea pig, 1:5. The sum of total CoQ9 and CoQ10 in rat and guinea-pig hepatocytes was about 780 and 400 pmol/mg protein, respectively. The concentration of CoQ9H2 in rat hepatocytes decreased linearly after the addition of AAPH, whereas that of oxidized CoQ9 showed a reciprocal increase. No loss of cell viability or increase of lipid peroxidation was observed until most of the CoQ9H2 had been consumed. Cellular CoQ9H2 was consumed probably through scavenging of lipid peroxyl radicals produced by incubation with AAPH. On the other hand, CoQ10H2 was not significantly consumed in the AAPH-treated rat hepatocytes during incubation compared with the control cells. In guinea-pig hepatocytes, cellular CoQ10H2 as well as CoQ9H2 was consumed by addition of AAPH. alpha-Tocopherol also showed linear consumption with incubation time regardless of the cell types used. It is concluded that CoQ9H2, together with alpha-tocopherol, constantly acts as a potential antioxidant in hepatocytes when incubated with AAPH, whereas CoQ10H2 mainly exhibits its antioxidant activity in cells containing CoQ10 as the predominant CoQ homolog.

Topics: Amidines; Animals; Antioxidants; Cell Survival; Cells, Cultured; Coenzymes; Guinea Pigs; Kinetics; Lipid Peroxidation; Liver; Male; Oxidation-Reduction; Rats; Rats, Inbred Strains; Solubility; Ubiquinone

Chronic gamma-irradiation of rats with the daily dose of 0.129 Gy activates the synthesis of various classes of lipids in the thymus, spleen and bone marrow cells and induces lipid accumulation in these tissues. Feeding of rats with the antioxidant, ubiquinone Q-9, under conditions of chronic irradiation causes a considerable normalization of lipogenesis and levels of the lipid concentration in the tissues of animals irradiated with the dose of 20 Gy.

Topics: Animals; Bone Marrow; Bone Marrow Cells; Cells, Cultured; Gamma Rays; Male; Phospholipids; Rats; Rats, Inbred Strains; Spleen; Thymus Gland; Ubiquinone

The influence of different kinds of dietary fat (8%) and of endogenous lipid peroxidation with regard to coenzyme Q9 (CoQ9) and coenzyme Q10 (CoQ10) concentrations in mitochondria and microsomes from rat liver has been investigated by means of an HPLC technique. Although the different diet fats used did not produce any effect on microsomes, it was possible to show that each experimental diet differently influenced the mitochondrial levels of CoQ9 and CoQ10. The highest mitochondrial CoQ content was found in case of a diet supplemented with corn oil. An endogenous oxidative stress induced by adriamycin was able to produce a sharp decrease in mitochondrial CoQ9 levels in the rats to which corn oil was administered. The results suggest that dietary fat ought to be considered when studies concerning CoQ mitochondrial levels are carried out.

Topics: Animals; Chromatography, High Pressure Liquid; Coenzymes; Corn Oil; Dietary Fats, Unsaturated; Doxorubicin; Lipid Peroxidation; Male; Malondialdehyde; Microsomes, Liver; Mitochondria, Liver; Olive Oil; Plant Oils; Rats; Rats, Inbred Strains; Ubiquinone

Effects of coenzyme Q9 (25 mg/kg), N6-cyclohexyl adenosine (CHA, 100 micrograms/kg) and their combination were compared in rats with short-term or permanent ligation of the left coronary artery. The following parameters were evaluated in three series of experiments: 1) incidence and duration of ventricular fibrillation and tachycardia during coronary occlusion (10 min) and consecutive reperfusion (5 min); 2) contractility and electrical stability of the heart (ventricular fibrillation threshold) in animals with 2-day myocardial infarction; 3) ischemic myocardial mass after coronary occlusion (5 min) and necrotic tissue mass in 2-day myocardial infarction. The rats were given oral drugs 5 days and 2 hours before the study. All the experiments were performed in open-chest anesthetized (nembutal, 50 mg/kg) rats exposed to ventilation at room air. Both the coenzyme Q9 and CHA significantly reduced the incidence and duration of coronary occlusion and reperfusion arrhythmias, prevented cardiac contractile depression (heart rate.developed pressure) and increased ventricular fibrillation threshold). The effect of coenzyme Q9 was more marked than that of CHA. Coenzyme Q9 substantially reduced necrotic tissue mass while CHA diminished ischemic tissue mass. At the same time the total cardioprotective action of the Q9 + CHA combination was more pronounced than that of them used alone.

Topics: Adenosine; Animals; Disease Models, Animal; Drug Evaluation, Preclinical; Heart Arrest, Induced; Male; Myocardial Infarction; Myocardial Reperfusion Injury; Rats; Rats, Inbred Strains; Ubiquinone

In order to determine whether coenzyme Q (CoQ) homologs which coexist in mammals play the same or different roles, the concentrations of coenzyme Q9 (CoQ9) and coenzyme Q10 (CoQ10) were analyzed in Japanese White (JW) rabbit tissues during growth, together with the intracellular distribution of these two CoQ homologs. In liver %CoQ9 (total [CoQ9] X 100/total [CoQ9] + total [CoQ10]) was approx. 40% until 3 weeks after birth, and then gradually decreased to 20%. In kidney, %CoQ9 decreased from 8% (1 week) to 1% (7 weeks). In heart, %CoQ9 was 3%, and in the brain, 2%, and these values did not change with growth. Most CoQ9 was present in the cytosolic fraction, whereas most CoQ10 was in the mitochondrial fraction. There was but minor change in the intracellular distribution of CoQ9 and CoQ10 in rabbit liver between 2 weeks and 7 weeks of age. These results suggest that CoQ9 and CoQ10 may play different roles in their physiological actions as antioxidant or component of the mitochondrial respiratory chain.

Topics: Animals; Brain; Cytosol; Kidney; Liver; Mice; Mice, Inbred ICR; Microsomes; Mitochondria; Myocardium; Rabbits; Rats; Rats, Inbred Strains; Ubiquinone

Topics: Animals; Antioxidants; Chromatography, High Pressure Liquid; Female; Mice; Mice, Hairless; Skin; Skin Aging; Skin Diseases; Ubiquinone; Ultraviolet Rays; Vitamin E

The long-chain fatty acid compositions of 22 species of Candida were determined, and compared with the fatty acid compositions of 10 species of the genus Pichia that contain coenzyme Q9. The long-chain fatty acid results were also compared with other phenotypic criteria (i.e. assimilation of carbon sources, coenzyme Q type, G + C content and proton magnetic resonance spectra) in order to establish possible anamorph/teleomorph relations. Close correlations were found between known perfect/imperfect states. The results suggest that C. cacaoi and P. farinosa, and C. maltosa and P. etchellsii, also have anamorph/teleomorph relationships.

Topics: Candida; Fatty Acids; Phenotype; Pichia; Saccharomycetales; Ubiquinone

A hitherto undescribed ballistosporous yeast was isolated from a dead leaf of Yucca sp. in Canada. This yeast produces apiculate or short-ellipsoidal ballistospores, produces pale colored colonies, has Q-9 as the major ubiquinone, and does not contain xylose in the cells. This new yeast is described as Sporobolomyces yuccicola Nakase et Suzuki. Sporobolomyces yuccicola is the sixth species of the intermedius group, a group of atypical species of the genus Sporobolomyces equipped with Q-9.

Topics: Chromatography, High Pressure Liquid; Culture Media; DNA, Fungal; Mitosporic Fungi; Spores, Fungal; Ubiquinone; Xylose

We measured ubiquinone (UQ)-9 and UQ-10 content in various foods using high performance liquid chromatography. UQ-9 was detected in cereals, some vegetables and their products. Corn oil and wheat germ had large amounts of UQ-9 in particular. UQ-10 was detected in meats, fishes, pulses, nuts, dairy products and various vegetables. Migratory fishes, rapeseed oil and soybean oil had considerably large amounts of UQ-10.

Topics: Animals; Edible Grain; Fabaceae; Fats; Fishes; Food Analysis; Nuts; Oils; Plants, Medicinal; Shellfish; Ubiquinone; Vegetables

The protective action of hepatic cells of the coenzyme CoQ10 was checked against the well-known hepatolesive agent carbon tetrachloride (CCl4). It was found that CoQ10 pretreatment strongly reduced the CCl4-induced lesions in rat liver. The most important of these lesions was a marked steatosis, together with focal necrosis, Kupffer-cell reaction and signs of phlogosis and fibroblastic proliferation. The protective effects of CoQ10 seemed to be dose-dependent. The variation of CoQ9 concentration in the liver was not significant at any of the doses used.

Topics: Animals; Carbon Tetrachloride; Coenzymes; Liver; Rats; Rats, Inbred Strains; Ubiquinone

The synthesis and phospholipid content in the liver, intestine and spleen in normal and irradiated rats administered ubiquinone-9 were studied with the use of 3H-serine. Ubiquinone markedly activated decarboxylation of phosphatidylserine and suppressed transformation of phosphatidylethanolamine to phosphatidylcholine in rat liver and spleen. The effect was also observed in the organs of irradiated animals. In rat intestine, administration of ubiquinone normalized a sharp gamma-irradiation-induced inhibition of transformation of phosphatidylcholine from phosphatidylethanolamine. The catabolism of phospholipids under the action of ubiquinone and radiation was inhibited in the liver and, on the contrary, was activated in radiosensitive organs.

Topics: Animals; Drug Evaluation, Preclinical; Gamma Rays; Intestinal Mucosa; Intestines; Liver; Male; Phospholipids; Radiation Injuries, Experimental; Rats; Rats, Inbred Strains; Spleen; Ubiquinone

Topics: Animals; Brugia; Dipetalonema; Dirofilaria immitis; Filarioidea; Onchocerca; Polyisoprenyl Phosphates; Terpenes; Ubiquinone

We studied the effects of Coenzyme Q10 (CoQ10) on DBA/2 mice inoculated with the M variant of encephalomyocarditis virus. The mice were treated as follows: 1) CoQ group (n = 49); CoQ10 1.0 mg (0.1 ml) X 2/day (0.1 mg/g/day), 2) control group (n = 55); sham-liquid 0.1 ml X 2/day. These treatments were intraperitoneally performed every day on days -1 to 12. In both groups, we determined the heart and serum contents of CoQ9 and CoQ10, which are the biologically active forms of CoQ in mice, in the mice killed on days 3-4 and 7. There was no significant change in the cumulative incidence of myocarditis in both groups. The survival rate was significantly higher on days 5-12 in the CoQ group than in the control group. There were significant increases of CoQ9 content on days 3-4, and CoQ10 content on days 3-4 and 7, in the heart in the CoQ group as compared with the control group. There was no significant change in the serum content of CoQ9 in both groups. The marked increase of the serum CoQ10 content seen in the CoQ group was due to the results of the exogenous administration of CoQ10. Thus, it may be concluded that CoQ10 may have a protective effect against viral myocarditis in man, in whom CoQ10 is only an active form of CoQ.

Topics: Animals; Coenzymes; Encephalomyocarditis virus; Enterovirus Infections; Mice; Mice, Inbred DBA; Myocarditis; Myocardium; Ubiquinone

Serum and tissue CoQ9 levels were determined in hypothyroid, euthyroid and hyperthyroid rats. A significant negative correlation was demonstrated between serum FT4 or T3 and CoQ9 in rats with various states of thyroid functions. Liver CoQ9 was significantly increased in rats rendered mildly hyperthyroid. There was a significant positive correlation between serum FT4 or T3 and liver CoQ9. While liver CoQ9 did not significantly change in severely hyperthyroid animals, liver mitochondrial CoQ9 showed a significant positive correlation with serum T3. Kidney and heart CoQ9 levels did not significantly change in hyperthyroid rats, but those in hypothyroid rats showed a tendency to increase. It was suggested that the synthesis of CoQ9 was increased in the liver in hyperthyroidism.

Topics: Animals; Kidney; Liver; Male; Mitochondria, Liver; Myocardium; Rats; Rats, Inbred Strains; Thyroid Diseases; Thyroxine; Triiodothyronine; Ubiquinone

A quantitative method for the determination of coenzyme Q10 (CoQ10) in human blood has been devised which allows recovery of essentially 100% of the CoQ10. The use of whole blood rather than plasma includes the CoQ10 in white cells. The method utilizes TLC instead of saponification to fractionate lipid impurities, because CoQ10 is sensitive to saponification, and utilizes CoQ11 as an internal standard which is advantageous over CoQ9 and a synthetic quinone. The final step of HPLC frequently reveals a peak with a retention time like that of CoQ9 which, being less than that of CoQ10, can be near other peaks of impurities.

Topics: Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Coenzymes; Humans; Leukocytes; Ubiquinone

To investigate the perturbation of ubiquinone biosynthesis by a hypocholesterolemic drug, 3 beta-(2-diethylaminoethoxy)androst-5-en-17-one hydrochloride (U18666A), we measured the incorporation of radioactive mevalonate, methionine, tyrosine, and 4-hydroxybenzoic acid into ubiquinone in glioblastoma cells. These four precursors unanimously showed that ubiquinone biosynthesis was not significantly altered by U18666A, which blocked cholesterol biosynthesis at steps beyond mevalonate formation. The fluctuation of the endogenous mevalonate level had little effect on ubiquinone biosynthesis, implying the relative stability of cellular ubiquinone biosynthesis. Furthermore, exogenously added mevalonate did not have an appreciable effect on ubiquinone biosynthesis. The major ubiquinone produced in rat glioblastoma cells was identified as ubiquinone-9. The mevalonate-derived products accumulated in the U18666A-treated cells differed significantly from those reported in a broken cell study, suggesting the existence of delicate mechanisms regulating the formation of cholesterol intermediates.

Topics: Androstenes; Animals; Anticholesteremic Agents; Cell Line; Glioma; Hydroxybenzoates; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Lovastatin; Methionine; Mevalonic Acid; Naphthalenes; Parabens; Rats; Tyrosine; Ubiquinone

The present study was undertaken to determine whether hepatic ischemia and the subsequent reflow of blood had any effect on the levels of endogenous coenzyme Q homologs, alpha-tocopherol, and glutathione, and whether coenzyme Q10 (6 mg/kg of body weight) altered these levels. Ischemia of the rat liver for 90 min resulted in decreases of 19.1 and 19.6% of endogenous alpha-tocopherol and total glutathione (GSH + GSSG) without significant changes in the levels of endogenous total coenzyme Q homologs (oxidized and reduced). Restoration of the blood flow resulted in marked decreases in endogenous coenzyme Q homologs, alpha-tocopherol, and total glutathione in the control group. In coenzyme Q10-treated animals, however, there were no changes in the levels of endogenous total coenzyme Q9, alpha-tocopherol, or total glutathione as well as in the level of the enhanced total coenzyme Q10 during the reperfusion period. On the other hand, decreases in alpha-tocopherol and total glutathione during the period of ischemia remained unchanged. These results are compatible with the assumption that cellular damage caused by hepatic ischemia can be explained by free radical reaction processes during ischemia and especially, reperfusion and suggest that exogenous coenzyme Q10 functions as an antioxidant with endogenous coenzyme Q homologs, alpha-tocopherol, and glutathione in lipid peroxidation during reperfusion.

Topics: Animals; Coenzymes; Glutathione; Ischemia; Liver; Male; Oxidation-Reduction; Rats; Rats, Inbred Strains; Ubiquinone; Vitamin E

The thermotropic properties of coenzymes Q10, Q9, Q8, and Q7 have been examined by differential scanning calorimetry and wide-angle X-ray diffraction. Typical scanning calorimetry cooling curves of coenzyme Q from the liquid state exhibit a single exothermic phase transition into a crystalline state at a temperature that decreases as the length of the polyisoprenoid side-chain substituent decreases. Upon subsequent heating, the molecules undergo a series of thermal events which precede the main crystalline-to-liquid endothermic phase transition. The temperature of these transitions increases with increasing chain length. The crystallization phase transition temperature depends markedly on the rate at which the sample is cooled and increases with decreasing scan rate; the temperature of the melting endotherm is not markedly affected by the scan rate. Detailed calorimetric studies of coenzyme Q10 indicate that two crystalline states are formed, one at relatively high cooling rates to low temperatures and the other when preparations are cooled slowly from the liquid state to relatively high temperatures. Heating the crystalline phase formed by rapid cooling causes its transformation into the phase observed by cooling slowly. X-ray diffraction analysis confirmed the existence of these two crystal phases in coenzymes Q9 and Q10 and the transformation from the rapidly crystallized form to the more ordered form associated with slower cooling rates. At body temperature (310 K) under equilibrium conditions coenzyme Q10 exists in an ordered crystalline phase; the implications of the thermotropic behavior of coenzyme Q10 on mitochondrial function in vitro and in vivo are discussed.

Topics: Calorimetry, Differential Scanning; Coenzymes; Crystallization; Crystallography, X-Ray; Mitochondria; Thermodynamics; Ubiquinone

Among various ubiquinone (Q) isoprenologues tested, only Q7 was more efficient than menadione in promoting the oxidation of 5-methyltetrahydrofolate (CH3FH4) by 5,10-methylenetetrahydrofolate reductase isolated from adult Brugia pahangi, whereas Q10 was the best cofactor in the same reaction catalysed by the analogous enzyme from adult Dirofilaria immitis. Menoctone (3-[1-cyclohexyloctyl] -2-hydroxy-1,4-naphthoquinone) was a strong competitive inhibitor of both these ubiquinone isoprenologues in the respective reactions. When incubated in the presence of D,L-[14C]-mevalonate, adult B. pahangi and D. immitis synthesized radiolabelled Q9 only, in addition to other isoprenoid derivatives in the neutral lipid fraction. In view of the inability of Q9 to promote the oxidation of CH3FH4 by 5,10-methylenetetrahydrofolate reductase from B. pahangi, it seems unlikely that this filaria uses Q9 as a cofactor in this reaction. Conceivably, D. immitis could use Q9 as a cofactor in its enzymatic oxidation of CH3FH4, since in this circumstance, it was a better cofactor than menadione.

Topics: Brugia; Dirofilaria; Filarioidea; Naphthoquinones; Tetrahydrofolates; Ubiquinone; Vitamin K

Topics: Animals; Anti-Infective Agents; Coenzymes; Female; Immunity, Cellular; Listeriosis; Mice; Spleen; Ubiquinone

Topics: Animals; Drug Evaluation, Preclinical; Gamma Rays; Male; Mice; Radiation Injuries, Experimental; Radiation-Protective Agents; Time Factors; Ubiquinone

Topics: Acetates; Amino Acids; Amino Acids, Aromatic; Carbon Isotopes; Chromatography; Liver; Metabolism; Pharmacology; Phenylalanine; Rats; Research; Tyrosine; Ubiquinone

Topics: Basidiomycota; Coenzymes; Fungi; Quinones; Ubiquinone

Topics: Coenzymes; Diet; Ubiquinone