linoleic-acid and Hemolysis

Lipid-modified low molecular weight branched polyethyleneimines (PEIs) are promising non-viral gene delivery systems that have been successfully explored for treatment of various diseases. The present study aims to determine in vitro safety of these delivery systems based on assessment of cytotoxicity with peripheral blood mononuclear cells (PBMCs), hemolysis with human red blood cells (RBC) and cytokine secretion from several sources of PBMCs. The viability of cells treated with lipopolymer/pDNA complexes was dependent on the polymer:pDNA ratio used but remained low at therapeutically relevant concentrations for most lipopolymers, except for the propionic acid substituted PEIs. The extent of hemolysis was minimal and below the accepted safety levels with most of the lipopolymers; however, some linoleic acid substituted PEIs yielded significant hemolysis activity. Unlike strong cytokine secretion from PMA/IO stimulated cells, most lipopolymer/pDNA complexes remained non-responsive, showing minimal changes in cytokine secretion (TNF-α, IL-6 and IFN-γ) irrespective of the lipopolymer/pDNA formulations. The 0.6 kDa PEI with lauric acid substituent displayed slight cytokine upregulation, however it remained low relative to the positive controls. This study demonstrated that the lipid modified LMW PEIs are expected to be safe in contact with blood components. However, close attention to lipopolymer concentration and ratio of polymer to pDNA in formulations might be required for individual lipopolymers for optimal safety response in nucleic acid therapies. STATEMENT OF SIGNIFICANCE: This manuscript investigated the safety aspects of various lipid modified low molecular weight polyethylenimine (LMW-PEI) polymers employed for pDNA delivery through in vitro studies. Using peripheral blood mononuclear cells (PBMCs) from multiple sources, we show that the hemolysis ability was minimal for most polymers, although a particular lipid substituent (linoleic acid) at specific ratios exhibited hemolysis. The levels of pro-inflammatory cytokines (TNF-α, IL-6 and IFN-γ) were slightly upregulated only with a lauric acid substituted 0.6PEI, but remained low relative to positive control treatments. We further report the beneficial effect of polyacrylic acid additives on hemolysis and cytokine secretion to a reasonable extent. This study confirms the feasibility of using LMW-PEI as safe delivery agents for various therapeutic purposes.

Topics: Gene Transfer Techniques; Hemolysis; Humans; Interleukin-6; Leukocytes, Mononuclear; Linoleic Acid; Molecular Weight; Plasmids; Polyethyleneimine; Transfection; Tumor Necrosis Factor-alpha

The aim of the present study was to prepare chitosan-PVA-silver nanoparticles (CS-AgNPs) through green method. Chitosan and PVA polymers acted as stabilizing agents. DLS and TEM analyses showed that CS-AgNPs were homogeneously dispersed in matrix with an average size of 190-200 nm. The CS-AgNPs were tested for their antioxidant and antibacterial properties and the results revealed that they exhibited higher antioxidant activity than CS powder. Moreover, CS-AgNPs were characterized by a low cytotoxicity effect at 5-200 μg/ml against Chinese Hamster Ovary (CHO-K1) cells. In addition, the prepared CS-Ag NPs were found to promote significantly the wound healing, as determined by the wound contraction ratio and histological examination. A significant improvement in wound healing progression and in oxidative stress damage were observed for CS, CS-PVA and CS-AgNPs-treated wound tissues, when compared to control and CICAFLORA®-treated groups. The wound healing effect could be attributed to the antibacterial and antioxidant synergy of AgNPs and CS. Results strongly support the possibility of using CS-AgNPs for wound care applications.

Topics: Animals; Anti-Bacterial Agents; Antioxidants; beta Carotene; Biphenyl Compounds; Chitosan; CHO Cells; Cricetinae; Cricetulus; Escherichia coli; Free Radical Scavengers; Free Radicals; Gels; Hemolysis; Humans; Iron; Linoleic Acid; Male; Metal Nanoparticles; Microbial Sensitivity Tests; Oxidative Stress; Oxygen; Picrates; Rats; Rats, Wistar; Silver; Silver Nitrate; Skin; Staphylococcus aureus; Wound Healing

The phytochemical screening, antimicrobial, antioxidant and cytotoxic properties of Camellia sinensis were evaluated in the present study. The phytochemical screening revealed the presence of an applicable amount of lycopene, β-carotenes, flavonoids and tannins in C. sinensis. Among the phytochemicals, tannin was found to be significantly higher in tea plant. The antimicrobial activity of plant extracts against selected bacterial strains namely, Escherichia coli, Staphylococcus aurous, Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, Marginella morganii and Haemophilus influenzae was investigated. The results showed that the stem part of C. sinensis presented greater antimicrobial potential than the leaf and root. Antioxidant activity (assessed through % inhibition of linoleic acid per oxidation test) was the highest (89.22%) in n-hexane extract of root part as compared to other extracts. Finally, the cytotoxicity analysis (haemolytic activity against human erythrocytes) of plant extract showed the negligible (%) lysis of RBCs ranging from 1.73 to 4.01%. In conclusion, it can be suggested that C. sinensis is the potential source to obtain bioactive phenolic compounds with high antimicrobial and antioxidant properties, which could possibly be exploited for the treatment of various infectious diseases.

Topics: Anti-Infective Agents; Antioxidants; Camellia sinensis; Cells, Cultured; Erythrocytes; Gram-Negative Bacteria; Gram-Positive Bacteria; Hemolysis; Humans; Linoleic Acid; Oxidation-Reduction; Plant Extracts; Plant Leaves; Plant Roots; Plant Stems; Solvents

A series of cyclen-based linear oligomers bearing hydrophobic long chains (lipopolymers Cy-LC, where Cy and LC represent cyclen-based linear backbone and hydrophobic long chain substituents, respectively) were designed and synthesized. The effects of type and degree of substitution (DS) of hydrophobic long chains on the transfection efficiency were systematically studied. The nitrogen atoms with relatively strong basicity on the cyclen ensure their good DNA binding ability, which was confirmed by gel retardation and ethidium bromide exclusion assays. Lipopolyplexes could be formed as nanoparticles with suitable sizes and zeta potentials for gene transfection. In vitro gene delivery experiments revealed that the linoleic acid (LIN) substituted material Cy-LIN has better transfection efficiency than 25 kDa polyethylenimine in the absence or in the presence of serum. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and hemolysis assays showed low cytotoxicity and good biocompatibility of the lipopolyplexes. Fluorescent labeled DNA was used to study the cellular uptake and intracellular distribution of transfected DNA. Flow cytometry results suggested that a long chain is necessary for efficient cellular uptake, and images from confocal laser scanning microscopy showed that after 4h transfection, most of the fluorescent labeled DNA accumulated in the perinuclear region, which was required for efficient gene expression. Moreover, it was also found that the DS of the hydrophobic moiety can adjust the balance between DNA binding ability and dissociation of polyplexes, significantly affecting the transfection efficiency.

Topics: Animals; Biocompatible Materials; Cell Death; Cell Line, Tumor; Cyclams; DNA; Electrophoresis, Agar Gel; Endocytosis; Flow Cytometry; Fluorescence; Gene Expression; Gene Transfer Techniques; HEK293 Cells; Hemolysis; Heterocyclic Compounds; Humans; Hydrophobic and Hydrophilic Interactions; Intracellular Space; Linoleic Acid; Lipids; Luciferases; Particle Size; Plasmids; Rabbits; Static Electricity; Transfection

Novel surfactants made of diglutamic acid (DG) polar head linked to lithocholic, arachidonic, linoleic or stearic acids were designed for drug solubilization.. Surfactants 3-D conformer and packing parameter were determined by molecular modelling and self-assembling properties by pyrene fluorescence measurements. Cytotoxicity was assessed on Human Umbilical Vein Endothelial Cells (HUVEC) and haemolyitic activity on rat red blood cells. Drug solubilization was quantified and its interaction with hydrophobic moieties was characterized using differential scanning calorimetry and X-ray diffraction. Self organisation of stearoyl-DG was observed by cryogenic transmission electron microscopy. Toxicity after repeated injections of stearoyl-DG was investigated in Wistar rats.. DG-based surfactants self-assemble into water and their critical micellar concentrations are comprised between 200 and 920 μg/mL. Cytotoxicity and haemolysis were lower than for polysorbate 80. At best, stearoyl-DG solubilized the drug up to 22% (w/w). Solid-state characterization evidenced drug/lipid interactions leading to the formation of a new complex. Stearoyl-DG formed spherical micelles of 20 nm, as predicted by packing parameter calculation. However, it induced a possible liver toxicity after intravenous administration in rats.. Among the surfactants tested, stearoyl-DG is the more efficient for drug solubilization but its use is limited by its possible liver toxicity.

Topics: Animals; Antineoplastic Agents; Arachidonic Acid; Erythrocytes; Glutamic Acid; Hemolysis; Human Umbilical Vein Endothelial Cells; Humans; Hydrophobic and Hydrophilic Interactions; Linoleic Acid; Lithocholic Acid; Micelles; Models, Molecular; Rats; Rats, Wistar; Solubility; Stearic Acids; Surface-Active Agents

Resveratrol (3,5,4'-trihydroxy-trans-stilbene) is a naturally occurring polyphenol widely distributed in food and dietary plants. This phytochemical has been intensively studied as an efficient antioxidant and anticancer agent, and a variety of substituted stilbenes have been developed in order to improve the potency of resveratrol. In this work, we described the synthesis of 3,4,4 -trihydroxy-trans-stilbene (3,4,4'-THS), an analogue of resveratrol, and studied its antioxidant and cytotoxic activity in vitro. 3,4,4 -THS was much more efficient than resveratrol in protecting against free radical-induced lipid peroxidation, photo-sensitized DNA oxidative damage, and free radical-induced hemolysis of human red blood cells. More potent growth inhibition in cultured human leukemia cells (HL-60) was also observed for 3,4,4 -THS. The relationship between the antioxidant efficiency and cytotoxic activity was discussed, with the emphasis on inhibition of the free radical enzyme ribonucleotide reductase by antioxidants. The result that this subtle structure modification of resveratrol drastically improves its bioactivity provides important strategy to develop novel resveratrol-based molecules.

Topics: Anisoles; Antineoplastic Agents; Antioxidants; Cell Proliferation; DNA Damage; Free Radicals; Hemolysis; HL-60 Cells; Humans; Kinetics; Linoleic Acid; Lipid Peroxidation; Molecular Structure; Oxidative Stress; Resveratrol; Stilbenes; Structure-Activity Relationship

Several studies indicated that bifidobacteria possessed strong antioxidant activity. In present study, the antioxidant activities of Bifidobacterium animalis 01 proteins were evaluated using six assays, namely, linoleic acid preoxidation assay, erythrocyte hemolysis assay, 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, reducing power assay, hydroxyl (.OH) and superoxide radicals (.O(2)(-)) assays, in which the last two assays were measured by electron spin resonance (ESR). There were two kinds of B. animalis 01 proteins in this study, the regular B. animalis 01 protein (Pro-CK) and the B. animalis 01 selenium-contained protein (Pro-Se). Both Pro-CK and Pro-Se showed concentration dependent antioxidant activity in DPPH assay, reducing power assay and erythrocyte hemolysis assay. All results of six assays indicated that the antioxidant activity of the B. animalis 01 protein was improved remarkably after selenium was incorporated. The antioxidant activity of Pro-Se increased with the increase of selenium content in Pro-Se suggesting selenium played a positive role in enhancing the antioxidant activity of B. animalis 01 protein. Moreover, organic selenium was more effective than inorganic selenium on enhancing the hydroxyl radical scavenging ability of B. animalis 01 protein.

Topics: Antioxidants; Bacterial Proteins; Bifidobacterium; Biphenyl Compounds; Hemolysis; Humans; Hydroxyl Radical; Linoleic Acid; Oxidation-Reduction; Picrates; Selenium; Superoxides

The bulk of LDL entrapped in the arterial intima is modified by hydrolytic enzymes, leading to extensive cleavage of cholesterylesters and liberation of fatty acids. The latter induce apoptosis in endothelial cells but are far less cytotoxic towards macrophages. We have compared the cytotoxic effects of enzymatically modified LDL (E-LDL) on macrophages and polymorphonuclear granulocytes (PMN).. E-LDL displayed toxicity towards PMN at far lower concentrations than towards monocyte-derived macrophages. Native or oxidized LDL had no effect. Free fatty acids contained in E-LDL were the cause of the observed toxicity, which could be mimicked by linoleic acid, oleic acid and arachidonic acid. E-LDL provoked Ca(2+) influx and activated PMN, as witnessed by the generation of superoxide anions and peroxidase secretion. Inhibition of either oxidative burst or calcium influx did not diminish the cytotoxicity of E-LDL. Similar to free linoleic acid, E-LDL lysed red blood cells and rapidly rendered cells permeable to propidium iodide.. Possibly through their capacity to directly perturb cell membranes, free fatty acids contained in E-LDL exert potent cytotoxic effects on PMN. This may be one reason why PMN are not abundantly present in atherosclerotic lesions, and why PMN-depletion suppresses atherogenesis.

Topics: Adenosine Triphosphate; Animals; Arachidonic Acid; Atherosclerosis; Calcium; Cell Death; Cell Membrane Permeability; Cell Survival; Cells, Cultured; Fatty Acids, Nonesterified; Hemolysis; Humans; Hydrolysis; L-Lactate Dehydrogenase; Linoleic Acid; Lipoproteins, LDL; Macrophages; Neutrophils; Oleic Acid; Peptide Hydrolases; Peroxidase; Rabbits; Respiratory Burst; Sterol Esterase; Superoxides; Time Factors

The photobiological properties of 4-acetylaminophenylacetic acid (Actarit, ACT, MS-932, CAS 18699-02-0) were studied using in vitro phototoxicity assays: photohemolysis, photoperoxidation of linoleic acid and Candida sp phototoxicity test. ACT reduced nitro blue tetrazolium (NBT) when irradiated with lambda > or = 300 nm in deoxygenated aqueous buffer solution (pH 7.4). The photohemolysis rate and photoperoxidation of linoleic acid were inhibited significantly by reduced glutathione. ACT was phototoxic to Candida sp. The isolation and identification of the photodegradation products of ACT in phosphate buffered saline solution (pH 7.4) and methanol were studied under aerobic conditions. Four compounds were identified and two of them isolated and characterized by spectroscopic methods. A photodecarboxylation with the participation of oxygen via a type I mechanism was proposed for the photodegradation of ACT which undergoes direct electron transfer from the excited state of ACT carboxylate and homolytic rupture of the alpha-carbon bond. A photodynamic mechanism involving radicals and electron transfer reactions was suggested for the observed in vitro phototoxicity.

Topics: Antirheumatic Agents; Candida; Dose-Response Relationship, Drug; Erythrocytes; Free Radicals; Hemolysis; Humans; In Vitro Techniques; Indicators and Reagents; Light; Linoleic Acid; Magnetic Resonance Spectroscopy; Microbial Sensitivity Tests; Peroxides; Phenylacetates; Photochemistry; Photosensitizing Agents; Spectrophotometry, Ultraviolet

Hydrocortisone 21-acetate (HCA) in methanol solution undergoes photodegradation under UVB light, as monitored by HPLC. Five main photoproducts have been isolated and characterized by means of NMR and mass spectroscopy. One of them derives from a Norrish I photoreaction which cleaves the C17-C20 bond of the steroid yielding the andro-derivative, a second product comes from a Yang-type photorearrangement which links C18 to C20 yielding a cyclobutane adduct. The former photoproduct, in turn, undergoes further photolysis giving rise to various photoproducts, of which three have been characterized. The first is a stereoisomer of the andro-derivative, the others arise from the opening of the five-membered ring. HCA also proved photounstable in the solid state and in a commercial formulation for topical use, thus confirming the requirements of the Pharmacopeias for light protection of this drug. Indeed, experiments on LPS-stimulated THP-1 cells demonstrated the loss of anti-inflammatory activity when HCA was UVB-photodegraded. The radical mechanism involved in HCA photolysis seems also responsible for the in vitro photohemolytic effect and lipid peroxidation induced by HCA in combination with UVB light.

Topics: Animals; Anti-Inflammatory Agents; Cell Differentiation; Cell Line, Tumor; Chromatography, High Pressure Liquid; Culture Media, Conditioned; Dose-Response Relationship, Drug; Dose-Response Relationship, Radiation; Erythrocytes; Hemolysis; Humans; Hydrocortisone; Interleukin-1beta; Leukemia, Monocytic, Acute; Linoleic Acid; Lipid Peroxidation; Macrophages; Mass Spectrometry; Mice; Molecular Structure; Nuclear Magnetic Resonance, Biomolecular; Photobiology; Photolysis; Tetradecanoylphorbol Acetate; Time Factors; Ultraviolet Rays

The antioxidant activities of curcumin, its natural demethoxy derivatives (demethoxycurcumin, Dmc and bisdemethoxycurcumin, Bdmc) and metabolite hydrogenated derivatives (tetrahydrocurcumin, THC; hexahydrocurcumin, HHC; octahydrocurcumin; OHC) were comparatively studied using 2,2-diphenyl-1-picrylhydrazyl (DDPH) radical, 2,2'-azobis(2-amidinopropane)dihydrochloride (AAPH) induced linoleic oxidation and AAPH induced red blood cell hemolysis assays. Hydrogenated derivatives of curcumin exhibited stronger DPPH scavenging activity compared to curcumin and a reference antioxidant, trolox. The scavenging activity significantly decreased in the order THC>HHC=OHC>trolox>curcumin>Dmc>>>Bdmc. Stronger antioxidant activities toward lipid peroxidation and red blood cell hemolysis were also demonstrated in the hydrogenated derivatives. By the model of AAPH induced linoleic oxidation, the stoichiometric number of peroxyl radical that can be trapped per molecule (n) of hydrogenated derivatives were 3.4, 3.8 and 3.1 for THC, HHC and OHC, respectively. The number (n) of curcumin and Dmc were 2.7 and 2.0, respectively, which are comparable to trolox, while it was 1.4 for Bdmc. The inhibition of AAPH induced red blood cell hemolysis significantly decreased in the order OHC>THC=HHC>trolox>curcumin=Dmc. Results in all models demonstrated the lower antioxidant activity of the demethoxy derivatives, suggesting the ortho-methoxyphenolic groups of curcumin are involved in antioxidant activities. On the other hand, hydrogenation at conjugated double bonds of the central seven carbon chain and beta diketone of curcumin to THC, HHC and OHC remarkably enhance antioxidant activity.

Topics: Amidines; Antioxidants; Biphenyl Compounds; Chromans; Curcumin; Diarylheptanoids; Erythrocyte Membrane; Free Radical Scavengers; Free Radicals; Hemolysis; Humans; Hydrogenation; In Vitro Techniques; Linoleic Acid; Lipid Peroxidation; Molecular Structure; Oxidants; Picrates; Structure-Activity Relationship; Time Factors

Aceclofenac (Airtal) (1) is a photoallergic and phototoxic anti-inflammatory and analgesic agent. This drug is photolabile under aerobic and anaerobic conditions. Irradiation of an ethanol-solution of aceclofenac under oxygen or argon at 290-320 nm affords via decarboxlation compound 2 as the main isolated and spectroscopically identified photoproduct, besides hydroxylamine derivates 3 and 4. A radical intermediate was evidenced spectrophotometrically with GSH and DTNB, as well as by the dimerization of cysteine. Red blood cell lysis photosensitized by 1-4 was investigated. Furthermore, in a lipid-photoperoxidation test with linoleic acid the in vitro phototoxicity of aceclofenac was also verified. The photoinduced generation of peroxide by compound 1 was determined during the irradiation in presence of NADPH by chemiluminescence. In relation to the photoallergic activity of this drug, the interaction of aceclofenac with human serum albumin (HSA) has been studied through fluorescence spectroscopy. No photoinduced binding was observed after irradiation of compounds 1 in the presence of human serum albumin.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Cysteine; Dermatitis, Phototoxic; Diclofenac; Erythrocytes; Glutathione; Hemolysis; Humans; In Vitro Techniques; Linoleic Acid; Lipid Peroxidation; Luminescence; Oxidants; Oxidation-Reduction; Oxygen; Photolysis; Serum Albumin; Ultraviolet Rays

Free radicals and other reactive oxygen species (ROS) are generated by all aerobic cells and are widely believed to play a significant role in aging as well as a number of degenerative or pathological diseases. This study compared the free radical-scavenging properties and antioxidant activity of YCP, a polysaccharide from the mycelium of a marine filamentous fungus Phoma herbarum YS 4108 and its two chemically sulfated derivatives YCP-S1 and YCP-S2. Sulfation, which masks hydroxyl groups of YCP polysaccharide molecule, could introduce new antioxidant activity, such as superoxide and hydroxyl radicals scavenging activity, metal chelating action, lipid peroxidation and linoleic acid oxidation inhibition capability. Furthermore, sulfated YCP was more potent than YCP at protecting erythrocytes against oxidative damage hemolysis. The current data suggest for the first time that sulfation of polysaccharide significantly increases its antioxidant activity and the chemical modification of polysaccharides may allow the preparation of derivatives with new properties and a variety of applications.

Topics: Animals; Antioxidants; Chelating Agents; Erythrocytes; Fungi; Hemolysis; Hydroxyl Radical; Linoleic Acid; Lipid Peroxidation; Polysaccharides; Rats; Spectroscopy, Fourier Transform Infrared; Sulfates; Superoxides

The photobiological properties of 6-methoxy-2-naphthylacetic acid (6-MNAA) were studied using a variety of in vitro phototoxicity assays: photohemolysis, photoperoxidation of linoleic acid, photosensitized degradation of histidine and thymine and the Candida phototoxicity test. 6-MNAA was phototoxic in vitro. 6-MNAA reduced nitro blue tetrazolium (NBT) when irradiated with lambda > or = 300 nm in deoxygenated aqueous buffer solution (pH 7.4). NBT can be reduced by reaction with the excited state of 6-MNAA subject to interference with molecular oxygen. The photohemolysis rate was inhibited by the presence of 1,4-diazabicyclo[2.2.2]octane (DABCO), sodium azide (NaN3) and reduced glutathione (GSH). Photoperoxidation of linoleic acid and photosensitized degradation of histidine and thymine were significantly inhibited by sodium azide and reduced glutathione. 6-MNAA was phototoxic to C. albicans, C. lipolytica and C. tropicalis. A mechanism involving singlet oxygen, radicals, and electron transfer reactions is suggested for the observed phototoxicity.

Topics: Aerobiosis; Anaerobiosis; Anti-Inflammatory Agents, Non-Steroidal; Butanones; Candida; Dermatitis, Phototoxic; DNA; Erythrocytes; Hemolysis; Histidine; Humans; In Vitro Techniques; Light; Linoleic Acid; Lipid Peroxidation; Nabumetone; Naphthaleneacetic Acids; Nitroblue Tetrazolium; Photochemistry; Photosensitizing Agents; Thymine

The antioxidant effect of strictinin (SOH), which was extracted from green tea leaves, against the peroxidation of linoleic acid in sodium dodecyl sulfate (SDS) and cetyl trimethylammonium (CTAB) micelles, against the peroxidation of low-density lipoprotein (LDL) and against oxidative hemolysis of human red blood cells (RBCs), has been studied. The peroxidation of linoleic acid and LDL, and oxidative hemolysis of RBCs were initiated thermally by a water-soluble azo initiator 2,2'-azobis(2-amidinopropane hydrochloride) (AAPH), and the reaction kinetics in micelles and LDL were monitored by uptake of oxygen. The synergistic antioxidant effect of SOH with alpha-tocopherol (Vitamin E) was also studied by following the decay kinetics of alpha-tocopherol. Kinetic analysis of the antioxidation process demonstrates that SOH, used either alone or in combination with alpha-tocopherol, is an effective antioxidant against lipid peroxidation, but its effects significantly depend on the reaction medium.

Topics: alpha-Tocopherol; Antioxidants; Cetrimonium Compounds; Erythrocytes; Hemolysis; Humans; Kinetics; Linoleic Acid; Lipid Peroxidation; Lipoproteins, LDL; Micelles; Oxygen Consumption; Phenols; Quaternary Ammonium Compounds; Sodium Dodecyl Sulfate; Tea; Water

The neuroleptic drug levomepromazine (1, previously known as methotrimeprazine) is photolabile under UV-A and UV-B light in aerobic conditions. Irradiation of a methanol solution of this drug produces one photoproduct, resulting from the oxidation of 1 to its sulfoxide parent. It is demonstrated that photodegradation occurs via type II mechanism involving irreversible trapping of self-photogenerated singlet molecular oxygen. 1 shows a photohemolytic effect on human erythrocytes and photoinducers lipid peroxidation.

Topics: Antipsychotic Agents; Dermatitis, Phototoxic; Electrons; Erythrocytes; Hemolysis; Histidine; Humans; In Vitro Techniques; Indicators and Reagents; Linoleic Acid; Lipid Peroxidation; Methotrimeprazine; Nitroblue Tetrazolium; Oxidants; Oxygen; Photochemistry; Photolysis; Serum Albumin; Singlet Oxygen; Ultraviolet Rays

Aloe-emodin (1), emodin (2) and rhein (3) were found to be photolabile by visible (390-500 nm) light under aerobic conditions. The drugs 1, 2 and 3 were phototoxic in vitro when examined by the photohemolysis test under both oxygen and argon atmospheres, although the photohemolysis rate was markedly lower under anaerobic conditions. The experiments were also carried out in the presence of butylated hydroxyanisole (BHA), reduced glutathione (GSH), sodium azide (NaN3) and superoxide dismutase (SOD). Based on the inhibition of this process on addition of BHA, GSH, SOD and NaN3, there would seem to be involvement of free radicals (type I mechanism) and singlet oxygen in the process (type II mechanism). The in vitro phototoxicity of this anthraquinone series was also verified in a lipid-photoperoxidation test with linoleic acid. In summary, this anthraquinone series is phototoxic in vitro. This behavior can be explained through the involvement of singlet oxygen and stable photoproducts.

Topics: Acetylcholinesterase; Aloe; Anthraquinones; Dermatitis, Phototoxic; Drug Stability; Emodin; Erythrocytes; Hemolysis; Humans; In Vitro Techniques; Linoleic Acid; Lipid Peroxidation; Photochemistry; Reactive Oxygen Species; Ultraviolet Rays

The photolability of nabumetone (NB, 1, 4-[6-methoxy-2-naphthalenyl]-2-butanone) and its photobiological properties were studied under aerobic and anaerobic conditions using a variety of in vitro phototoxicity assays: photohemolysis, photoperoxidation of linoleic acid, and photosensitized degradation of histidine and thymine. The photodegradation rate of NB in methanol and phosphate buffered saline (PBS) was enhanced under oxygenated media. NB was phototoxic in vitro. The photohemolysis rate was enhanced by deuterium oxide and inhibited by the presence of 1,4-diazabicyclo[2.2.2]octane (DABCO), butylated hydroxyanisole (BHA), sodium azide (NaN3) and reduced gluthathione (GSH). The induced photoperoxidation of linoleic acid was inhibited significantly by sodium azide and reduced gluthathione. Histidine and thymine were photodegraded by a photosensitized reaction induced by NB. A mechanism involving singlet oxygen, radicals and photoproducts is suggested for the observed photoxicity.

Topics: Aerobiosis; Anaerobiosis; Anti-Inflammatory Agents, Non-Steroidal; Butanones; Dermatitis, Phototoxic; Erythrocytes; Hemolysis; Histidine; Linoleic Acid; Lipid Peroxidation; Lipid Peroxides; Nabumetone; Photochemistry; Spectrophotometry, Ultraviolet; Thymine

The phototoxic antidiabetes drug glipizide (1) is photolabile under aerobic conditions and UV-B light. Irradiation of a phosphate-buffered solution of 1 under oxygen atmosphere produces 4 photoproducts as well as singlet oxygen, which was detected by trapping it with 2,5-dimethylfuran and by the histidine test. The photochemistry of 1 involves cleavage of the sulfonamine and the sulfonamine-R bonds. Red blood cell lysis, photosensitized by glipizide and the products of its aerobic photolysis were demonstrated. The photohemolysis rate was lower for 1 than for its photoproducts. Inhibition of this process on addition of 1, 4-diazabicyclo[2.2.2]octane (DABCO), reduced glutathione (GSH), Vitamin C, sodium azide, superoxide dismutase, and a-tocopherol confirmed the possibility of singlet oxygen, superoxide ion and free radicals participation. Furthermore, in a lipid-photoperoxidation test with linoleic acid the in vitro phototoxicity of glipizide was also verified. A low decreasing cell viability of lymphocytes and neutrophils was observed.

Topics: Cell Survival; Cells, Cultured; Erythrocytes; Free Radical Scavengers; Glipizide; Hemolysis; Humans; Hypoglycemic Agents; Kinetics; Linoleic Acid; Lipid Peroxidation; Lymphocytes; Neutrophils; Oxygen; Photolysis; Photosensitizing Agents; Radiation-Protective Agents; Reactive Oxygen Species; Singlet Oxygen; Spectrophotometry; Ultraviolet Rays

Amitriptyline and imipramine, two tricyclic antidepressant drugs, have been studied to evaluate their phototoxic potential using various models. Reactive oxygen species production was investigated. A negligible production of singlet oxygen was observed for both compounds whereas a significant production of superoxide anion was noted for amitriptyline in particular. Moderate red blood cell lysis under UVA light (365 nm) was induced in the presence of the two drugs at a concentration of 50 microM. Cellular phototoxicity was investigated on a murine fibroblast cell line (3T3). The two drugs were phototoxic causing cell death at a concentration of 100 microM and a UVA dose in the range of 3.3-6.6 J/cm2. Furthermore, the two drugs photosensitized the peroxidation of linoleic acid, as monitored by the formation of dienic hydroperoxides. The presence of BHA and GSH, two free radical scavengers, significantly reduced the lipid oxidation photoinduced by the drugs, suggesting a predominant involvement of radical species. Finally, the involvement of nucleic acids in the phototoxicity mechanism was also investigated using a pBR322 plasmid DNA as a model.

Topics: Amitriptyline; Animals; Antidepressive Agents, Tricyclic; Cells, Cultured; Chromatography, High Pressure Liquid; Coloring Agents; Dermatitis, Phototoxic; DNA; DNA Damage; Erythrocytes; Fibroblasts; Hemolysis; Imipramine; Linoleic Acid; Lipid Peroxidation; Mice; Mice, Inbred BALB C; Photochemistry; Reactive Oxygen Species; Tetrazolium Salts; Thiazoles; Ultraviolet Rays

Antioxidant activities of caffeoyltryptophan were investigated by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging system, the superoxide anion generation system and the superoxide anion-mediated linoleic acid peroxidation system. At 10 microM, caffeoyltryptophan showed greater scavenging activity on DPPH than dl-alpha-tocopherol or ascorbic acid. DPPH radical scavenging activity of caffeoyltryptophan increased dose-dependently at concentrations ranging from 1 to 50 microM; 1 mol of caffeoyltryptophan reacted with ca 4 mol of radical. Caffeoyltryptophan caused 80% inhibition of superoxide anion generation at 50 microM. The inhibitory activity of caffeoyltryptophan was as strong as that of 5-caffeoylquinic acid. Caffeoyltryptophan inhibited the formation of conjugated diene from linoleic acid. The inhibitory activity increased in the order caffeic acid < 5-caffeoylquinic acid < caffeoyltryptophan < dl-alpha-tocopherol. Effects on the in vitro haemolysis and peroxidation of mouse erythrocytes induced by H2O2 were also examined. Caffeoyltryptophan exhibited strong inhibitory activities; Tryptophan was ineffective in these systems. These data suggest that caffeoyltryptophan may be a natural antioxidant in the human diet and, as such, may intervene in toxicological processes that are mediated by radical mechanisms.

Topics: Animals; Antioxidants; Caffeic Acids; Free Radical Scavengers; Hemolysis; Linoleic Acid; Lipid Peroxidation; Male; Mice; Tryptophan

Carprofen (1a) is a photosensitizing nonsteroidal anti-inflammatory drug. It undergoes photodehalogenation from its triplet excited state. The resulting aryl radical (II) is able to abstract hydrogen atoms from model lipids, mediating their peroxidation by a type I mechanism. This aryl radical intermediate appears to be responsible for the observed photobiological effects of carprofen. The active involvement of the triplet state has been confirmed by direct detection of this species in laser flash photolysis and by quenching experiments with cyclohexadiene and naphthalene. Carprofen also photosensitizes singlet oxygen production with a quantum yield of 0.32. A minor reaction pathway is photodecarboxylation, which occurs from the excited singlet state and leads to an acetyl derivative (1b). In the case of the dehalogenated photoproduct (2a), photodecarboxylation to the ethyl (2d) and acetyl (2b) derivatives, together with singlet oxygen production (quantum yield = 0.18), is also possible. However, the biological activity of 2a in the linoleic acid photoperoxidation and photohemolysis tests is markedly lower than that of 1a, which constitutes further evidence in favor of the important role of photodehalogenation in the adverse photobiological effects of carprofen.

Topics: Carbazoles; Energy Transfer; Erythrocytes; Fluorescence; Hemolysis; Humans; Lasers; Linoleic Acid; Lipid Peroxidation; Luminescent Measurements; Photolysis; Photosensitizing Agents; Spectrophotometry, Ultraviolet

Two experiments were conducted to determine the effect of dietary linoleic to linolenic acid (LO:LN) ratio and dl-alpha-tocopheryl acetate (TA) supplementation on selected characteristics of the liver and cerebellum and on vitamin E status of turkey poults from hatch through 22 d of age. In Experiment 1, 1-d-old poults were fed diets containing no supplemental TA (0E) or 150 IU TA/kg diet (150E). Poults fed the 150E diet had greater (P < 0.001) concentrations of alpha-tocopherol (TOC) in the liver and plasma than those fed the 0E diet from 7 to 22 d of age. The 150E diet, however, did not completely overcome the decrease in liver and plasma TOC concentrations observed at these ages. The 150E diet had no effect on poult BW, feed efficiency, or on the weight, protein, lipid, or fatty acid concentrations of the liver. Thiobarbituric acid reactive substances assay of liver and hemolysis assay of red blood cells (RBC) showed that the 150E diet decreased the susceptibility of liver and RBC to in vitro peroxidation at 13 and 22 d of age. In Experiment 2, 1-d-old poults were fed the 0E and 150E diets in a complete factorial arrangement with decreasing ratios of LO:LN (10, 5, and 1). Dietary LO:LN ratio had no effect on RBC hemolysis or cerebellum TOC concentration. As the ratio of LO:LN decreased, the arachidonic acid content of liver and cerebellum lipids decreased. Ratios of n-6 to n-3 fatty acids in liver and cerebellum were directly related to dietary LO:LN at 13 and 22 d of age.

Topics: alpha-Linolenic Acid; alpha-Tocopherol; Animals; Arachidonic Acids; Cerebellum; Diet; Erythrocytes; Fatty Acids, Omega-3; Fatty Acids, Omega-6; Fatty Acids, Unsaturated; Food, Fortified; Hemolysis; Linoleic Acid; Linoleic Acids; Lipid Metabolism; Lipids; Liver; Male; Random Allocation; Thiobarbituric Acid Reactive Substances; Tocopherols; Turkeys; Vitamin E

The purpose of this study was to investigate the role of hydrolysis products of linoleic acid anilide (LAA), i.e., aniline and linoleic acid (LA), in the toxicity to the hemopoietic system, especially to the spleen. To achieve this, the parent compound (LAA) and its putative hydrolysis products, i.e., aniline or linoleic acid (LA), were given to male SD rats at equimolar doses (0.7 mmol/kg) in 0.25 ml mineral oil by gavage, daily, for 14 days. The controls received equal volumes of vehicle only. Five animals from each group were euthanized at Days 1, 7, and 28 following the last dose. At all time points, spleen weights increased in the LAA- and aniline-treated rats, but spleen to body weight ratios were increased only at Days 1 and 7 in these groups. No changes were observed in the LA-treated rats at any time point. RBC counts were decreased in the LAA and aniline groups at Days 1 and 7, whereas hemoglobin content was decreased by 20 and 13% in the LAA- and aniline-treated rats, respectively, only at Day 1. Methemoglobin content in the LAA and aniline groups also increased by 76 and 101%, respectively, at Day 1. Serum transaminases (AST and ALT) decreased in the LAA, aniline, and LA groups but the decreases were more consistent in the LA group. Serum IgA increased in the LAA and aniline groups only at Day 1. Splenic iron content was increased 381, 486, and 51% in the LAA-treated rats and 474, 491, and 58% in the aniline-treated rats at Days 1, 7, and 28, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Anilides; Aniline Compounds; Animals; Blood Cell Count; Hematopoiesis; Hemolysis; Immunoglobulins; Iron; L-Lactate Dehydrogenase; Linoleic Acid; Linoleic Acids; Male; Organ Size; Rats; Rats, Sprague-Dawley; Spleen; Transaminases

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Cell Line; Cell Membrane; Cells, Cultured; Fibroblasts; Hemolysis; Humans; Ketoprofen; Linoleic Acid; Linoleic Acids; Lipid Peroxidation; Liver; Molecular Conformation; Photochemistry; Photolysis; Rats; Rats, Sprague-Dawley; Ultraviolet Rays

Electric field pulses used for cell manipulation can cause irreversible cell damage. The mechanisms of the processes leading to such cell damage are very complicated. Our work demonstrated that exponential electric pulses with intensity of 2-7.5 kV/cm and duration of 5.2 ms were able to initiate peroxidation of fatty acid emulsions, liposomal membranes, red blood and Ehrlich ascite tumor cells. Electric pulses-induced peroxidation of erythrocyte membranes was followed by hemolysis. The electric treatment caused damage of E. coli membrane lipids which was accompanied by decreased cell survival. All these effects depended on field intensity. A relatively good correlation between pulse-induced peroxidation of erythrocyte membranes and hemolysis was observed. These results suggest that free radical mediated processes as lipid peroxidation and/or lipid degradation or fragmentation may be possible causes for electric pulses-induced irreversible cell damage.

Topics: Animals; Carcinoma, Ehrlich Tumor; Electroporation; Emulsions; Erythrocyte Membrane; Escherichia coli; Free Radical Scavengers; Hemolysis; In Vitro Techniques; Kinetics; Linoleic Acid; Linoleic Acids; Lipid Peroxidation; Liposomes; Membrane Lipids; Membranes, Artificial; Mice; Oxidation-Reduction; Rats; Thiobarbituric Acid Reactive Substances; Tumor Cells, Cultured

Red blood cell (RBC) negative charges and resistance to linoleic acid (LNA)-induced lysis were studied in Plasmodium yoelii-infected mice and in malaria (P. falciparum)-affected individuals. RBCs from mice infected with P. yoelii showed a progressive decrease in the net surface negative charges at 24 h after infection, reaching a minimal value on day 3, followed by a second phase that was characterised by a recovery to normal levels on day 6. Resistance to linoleic acid follows similar kinetics. These alterations preceded the appearance of parasites in the peripheral blood. A similar increase in LNA-induced lysis was observed in RBCs from malaria-affected individuals. These early membrane alterations of uninfected RBCs could be responsible for spreading of infection and RBC lysis during infection.

Topics: Adult; Animals; Electrochemistry; Electrophoresis; Erythrocyte Membrane; Erythrocytes; Female; Hemolysis; Humans; In Vitro Techniques; Linoleic Acid; Linoleic Acids; Malaria; Malaria, Falciparum; Male; Mice; Mice, Inbred BALB C; Middle Aged; Plasmodium yoelii

The phototoxic anti-hyperlipoproteinemic drug fenofibrate was found to be photolabile under aerobic and anaerobic conditions. Irradiation under argon of a methanol solution of this drug produced the photoproducts isopropyl 4-(1-[4-chlorophenyl]-1,2-dihydroxy)ethylphenoxyisobutyrate, 1,2-bis(4-chlorophenyl)-1,2bis (4-[isopropoxycarbonylisopropoxy]phenyl)ethane-1,2-diol and 4-(4-chlorobenzoyl)phenol, while under oxygen the photoproducts were 4-chloroperbenzoic acid, methyl 4-chlorobenzoate, 4-chlorobenzoic acid and singlet oxygen, as evidenced by trapping with 2,5-dimethylfuran. These results can be rationalized through hydrogen abstraction by excited fenofibrate, to afford a free radical as key intermediate. Biologically active antioxidants such as glutathione and cysteine efficiently reduced 4-chloroperbenzoic acid to 4-chlorobenzoic acid. The involvement of an electron transfer mechanism is suggested by detection (UV-vis spectrophotometry) of the radical cation TMP+. during the oxidation of tetramethylphenylenediamine (TMP) with 4-chloroperbenzoic acid. Fenofibrate was phototoxic in vitro when examined by the photohemolysis test, both under oxygen and argon atmosphere, although the photohemolysis rate was markedly lower under anaerobic conditions. The photoproducts 4-(1-[4-chlorophenyl]-1,2-dihydroxy)ethylphenoxyisobutyrate and 4-chloroperbenzoic acid induced hemolysis in the dark; however, this effect was quantitatively less important than photohemolysis by fenofibrate. On the other hand, fenofibrate photosensitized peroxidation of linoleic acid, monitored by the UV detection of dienic hydroperoxides. Based on the inhibition of this process upon addition of butylated hydroxyanisole, a radical chain (type I) mechanism appears to operate. In summary, fenofibrate is phototoxic in vitro. This behavior can be explained through the involvement of free radicals, singlet oxygen and stable photoproducts.

Topics: Erythrocytes; Fenofibrate; Hemolysis; Humans; Linoleic Acid; Linoleic Acids; Lipid Peroxidation; Photolysis; Photosensitizing Agents; Spectrophotometry

Vitamin E status was assessed in 22 patients with cystic fibrosis and 9 controls by measuring concentrations of the vitamin, vitamin E:lipid ratios and peroxide-induced haemolysis in plasma and erythrocytes. For a given concentration of plasma or erythrocyte alpha-tocopherol, erythrocytes of patients with cystic fibrosis were more susceptible to peroxide-induced haemolysis than controls. This susceptibility should be countered by supplementation with vitamin E to maintain higher than normal concentrations of circulating alpha-tocopherol-greater than 4.8 mmol alpha-tocopherol/mol cholesterol.

Topics: Adolescent; Arachidonic Acid; Child; Child, Preschool; Cholesterol; Cystic Fibrosis; Erythrocytes; Female; Hemolysis; Humans; Linoleic Acid; Linoleic Acids; Lipids; Male; Peroxides; Triglycerides; Vitamin E

Vitamin E and linoleate, both of which are found in high concentrations in sunflower seed oil, were examined independently for their influence on general and blood-vascular parameters in vitamin E-deficient common marmosets. A vitamin E-deficient diet (-E, 4 micrograms/g) was supplemented with either 40 micrograms/g vitamin E (+E), vitamin E stripped sunflower oil (+10% SSO-E), or SSO (+10% SSO w/w) in a 2 x 2 factorial designed experiment, and the diets fed for 9 months to 4 even groups of common marmosets. Vitamin E deficiency was associated in marmosets with a loss of skeletal muscle mass and of body weight, enhanced peroxidative haemolysis of erythrocytes, increased white blood cell counts, and in the SSO-E group a relative neutrophilia. Platelet reactivity was increased with vitamin E deficiency, and to a greater degree with the SSO-E group. Aortic prostacyclin production was significantly increased by the addition of vitamin E, linoleate and both as SSO to the deficient diet, the effects being additive. Fatty acid changes associated with the different treatments reflected the influence of high linoleate and vitamin E treatments. The platelet and aortic arachidonate value in the SSO-E group showed the lowest and most variable value, and this was associated with greatest platelet aggregability. An adequate vitamin E intake is essential for stabilising high PUFA diets and biomembranes and enhancing the protective role of prostacyclin in blood vessels against thrombogenesis.

Topics: Animals; Aorta; Blood Platelets; Body Weight; Callithrix; Dietary Fats, Unsaturated; Fatty Acids; Female; Hemolysis; Leukocyte Count; Linoleic Acid; Linoleic Acids; Lipid Peroxides; Muscles; Organ Size; Plant Oils; Platelet Aggregation; Prostaglandins; Prostaglandins F; Sunflower Oil; Thromboxane B2; Vitamin E; Vitamin E Deficiency

Purified rat hemoglobin catalyzes the oxidative degradation of iodothyronines to form iodide and an iodine-containing intermediate that reacts with protein. Hemoglobin also catalyzes peroxidation of linoleic acid. These observations are consistent with the reported intrinsic peroxidase activity of hemoglobin and other heme-proteins. However, incubations containing both linoleic acid and an iodothyronine produced a surprising result: deiodination was stimulated rather than competitively inhibited. In contrast, linoleic-acid peroxidation was inhibited by iodothyronines. Thus, low levels of iodothyronines (2.6 X 10(-7) M) are effective inhibitors of linoleic-acid peroxidation. Thyroxine and reverse T3 were found to be more effective in this antioxidant activity than vitamin E, glutathione, ascorbic acid and DTT. Since linoleic-acid peroxidation proceeds by a propagating free-radical mechanism, we have concluded that iodothyronines can effectively terminate the free-radical chain reaction to become oxidatively deiodinated. Consistent with this antioxidant mechanism, reverse T3 is effective in preserving red cell membranes as measured by the inhibition of erythrocyte hemolysis.

Topics: Animals; Antioxidants; Erythrocytes; Fatty Acids; Female; Hemoglobins; Hemolysis; In Vitro Techniques; Iodine; Kinetics; Linoleic Acid; Linoleic Acids; Lipid Peroxides; Oxidation-Reduction; Rats; Rats, Inbred Strains; Thyronines

Topics: Chromatography; Fatty Acids; Fatty Acids, Essential; Hemolysis; Linoleic Acid; Oleic Acid; Oleic Acids; Osmosis; Palmitic Acid; Research; Sodium Chloride; Stearic Acids

Topics: Butter; Castor Oil; Chromatography; Cocos; Dietary Fats; Electrolytes; Erythrocytes; Fatty Acids; Fatty Acids, Essential; Glycerol; Glycols; Hemolysis; Linoleic Acid; Lipids; Oils; Permeability; Pharmacology; Phospholipids; Rats; Research; Thiourea; Zea mays

Topics: Abetalipoproteinemia; Adenine Nucleotides; Adenosine Triphosphate; Autoimmune Diseases; Blood Protein Disorders; Celiac Disease; Cholesterol; Congenital Abnormalities; Erythrocytes; Glucosephosphate Dehydrogenase; Hemoglobinometry; Hemolysis; Linoleic Acid; Lipid Metabolism; Lipoproteins; Metabolism; Methemoglobin; Neurologic Manifestations; Osmosis; Phospholipids; Research