4-hydroxy-2-nonenal and 4-hydroxy-2-hexenal

The process of lipid oxidation generates a diverse array of small aldehydes and carbonyl-containing compounds, which may occur in free form or esterified within phospholipids and cholesterol esters. These aldehydes mostly result from fragmentation of fatty acyl chains following radical oxidation, and the products can be subdivided into alkanals, alkenals (usually α,β-unsaturated), γ-substituted alkenals and bis-aldehydes. Isolevuglandins are non-fragmented di-carbonyl compounds derived from H

Topics: Acrolein; Aldehydes; Animals; Humans; Isoprostanes; Lactoglobulins; Lipid Peroxidation; Oxidative Stress; Protein Processing, Post-Translational

4-Hydroxy-2E-hexenal (4-HHE) and 4-hydroxy-2E-nonenal (4-HNE) have been characterized as prominent by-products of n-3 and n-6 hydroperoxy derivatives of n-3 and n-6 fatty acids, respectively. We also have characterized the homolog 4-hydroxy-2E,6Z-dodecadienal (4-HDDE) as a specific by-product of the 12-lipoxygenase product of arachidonic acid 12-hydroperoxy-eicosatetraenoate (12-HpETE). The three hydroxy-alkenals have been found in human plasma with 4-HHE being the most prominent followed by 4-HNE. They were found increased in tissues submitted to oxidative stress, according to the fatty acid characteristic of those tissues, e.g., 4-HNE and 4-HDDE in blood platelets and 4-HHE in the retina. We have shown they covalently bind to the primary amine moiety of ethanolamine phospholipids (PE), especially the plasmalogen subclass, with the highest hydrophobic alkenal (4-HDDE) being the most reactive. Their carboxylic acid metabolites, 4-hydroxy-2E-hexenoic acid (4-HHA), 4-hydroxy-2E-nonenoic acid (4-HNA) and 4-hydroxy-2E,6Z-dodecadienoic acid (4-HDDA), respectively, were found in human urine and measured in higher amounts in situations in which oxidative stress has been reported such as aging and diabetes. As reported above with their hydroxy-alkenals precursors, 4-HHA proved to be the most prominent followed by 4-HNA. Altogether, the three hydroxy-alkenals, either in their free form or bound to membrane PE, may be considered as specific markers of lipid peroxidation able to discriminate between n-3 and n-6 fatty acids. This is corroborated by the measurement of their urinary carboxylic acid metabolites.

Topics: Aldehydes; Fatty Acids; Fatty Acids, Omega-3; Fatty Acids, Omega-6; Humans; Lipid Peroxidation; Molecular Structure; Phosphatidylethanolamines

Intake of fish oil reduces the risk of CHD and CHD deaths. Marine n-3 fatty acids (FA) are susceptible to oxidation, but to our knowledge, the health effects of intake of oxidised fish oil have not previously been investigated in human subjects. The aim of the present study was to investigate markers of oxidative stress, lipid peroxidation and inflammation, and the level of plasma n-3 FA after intake of oxidised fish oil. In a double-blinded randomised controlled study, healthy subjects (aged 18-50 years, n 54) were assigned into one of three groups receiving capsules containing either 8 g/d of fish oil (1.6 g/d EPA+DHA; n 17), 8 g/d of oxidised fish oil (1.6 g/d EPA+DHA; n 18) or 8 g/d of high-oleic sunflower oil (n 19). Fasting blood and morning spot urine samples were collected at weeks 0, 3 and 7. No significant changes between the different groups were observed with regard to urinary 8-iso-PGF2α; plasma levels of 4-hydroxy-2-hexenal, 4-hydroxy-2-nonenal and α-tocopherol; serum high sensitive C-reactive protein; or activity of antioxidant enzymes in erythrocytes. A significant increase in plasma level of EPA+DHA was observed in both fish oil groups, but no significant difference was observed between the fish oil groups. No changes in a variety of in vivo markers of oxidative stress, lipid peroxidation or inflammation were observed after daily intake of oxidised fish oil for 3 or 7 weeks, indicating that intake of oxidised fish oil may not have unfavourable short-term effects in healthy human subjects.

Topics: Adult; Aldehydes; alpha-Tocopherol; Biomarkers; C-Reactive Protein; Cod Liver Oil; Dietary Supplements; Dinoprost; Double-Blind Method; Erythrocytes; Fatty Acids, Omega-3; Female; Humans; Lipid Peroxidation; Male; Norway; Oxidation-Reduction; Oxidative Stress; Oxidoreductases; Patient Dropouts; Young Adult

Topics: Aldehydes; Chromatography, High Pressure Liquid; Fats, Unsaturated; Fatty Acids; Hot Temperature; Oxidation-Reduction; Plant Oils; Tocopherols

CKD is associated with increased oxidative stress that correlates with occurrence of cardiovascular events. Modifications induced by increased oxidative stress particularly affect circulating lipoproteins such as HDL that exhibit antiatheromatous and antithrombotic properties. To explore the specific role of oxidative modifications of HDL in CKD and their effect on the platelet-targeting antiaggregant properties of HDL, we used a CKD (5/6 nephrectomy) rabbit model. For. HDL from CKD rabbits and patients on hemodialysis had HNE adducts. The percentage of platelet aggregation or activation induced by collagen was significantly higher when platelets were incubated with HDL from CKD rabbit and hemodialysis groups than with HDL from the control group. In both rabbits and humans, platelet aggregation and activation were significantly higher in the presence of HNE-modified HDL than with HDL from their respective controls. Incubation of platelets with a blocking antibody directed against CD36 or with a pharmacologic inhibitor of SRC kinases restored the antiaggregative phenotype in the presence of HDL from CKD rabbits, patients on hemodialysis and peritoneal dialysis, and HNE-modified HDL.. HDL from CKD rabbits and patients on hemodialysis exhibited an impaired ability to inhibit platelet aggregation, suggesting that altered HDL properties may contribute to the increased cardiovascular risk in this population.

Topics: Adult; Aged; Aged, 80 and over; Aldehydes; Animals; Antibodies; Blood Platelets; CD36 Antigens; Cells, Cultured; Disease Models, Animal; Female; Humans; JNK Mitogen-Activated Protein Kinases; Lipoproteins, HDL; Male; Malondialdehyde; Middle Aged; Oxidation-Reduction; Oxidative Stress; Peritoneal Dialysis; Phosphorylation; Platelet Aggregation; Protein Carbonylation; Protein Kinase Inhibitors; Rabbits; Renal Insufficiency, Chronic; src-Family Kinases

Lipid aldehydes originating from the peroxidation of n-3 and n-6 polyunsaturated fatty acids are increased in hemodialysis (HD) patients, a process already known to promote oxidative stress. However, data are lacking for patients with chronic kidney disease (CKD) before the initiation of HD. We prospectively evaluated the changes of plasma concentrations of two major lipid aldehydes, 4-HHE and 4-HNE, according to the decrease of glomerular filtration rate (GFR) in 40 CKD and 13 non-CKD participants. GFR was measured by inulin or iohexol clearance. Thus, 4-hydroxy-2-nonenal (4-HNE) and 4-hydroxy-2-hexenal (4-HHE) were quantitated in plasma by gas chromatography coupled with mass spectrometry and their covalent adducts on proteins were quantified by immunoblotting. On the one hand, 4-HHE plasma concentration increased from CKD stage I-II to CKD stage IV-V compared to non-CKD patients (4.5-fold higher in CKD IV-V,

Topics: Adult; Aged; Aldehydes; Biomarkers; Case-Control Studies; Female; Gas Chromatography-Mass Spectrometry; Glomerular Filtration Rate; Humans; Kidney; Lipid Peroxidation; Male; Middle Aged; Oxidative Stress; Renal Insufficiency, Chronic; Up-Regulation

Muscle food characteristics (fatty acid profile, heme-Fe, intrinsic antioxidants) that relate to the formation of (patho)physiological oxidation products during gastrointestinal digestion are investigated.. Muscles (n = 33) from 18 mammal, poultry, and fish species, of which some are mixed with lard to standardize their fatty acid profile, are digested in vitro. Lipid oxidation is assessed by thiobarbituric reactive substances (TBARS), n-3 PUFA derivative 4-hydroxy-2-hexenal and propanal, n-6 PUFA derivative 4-hydroxy-2-nonenal and hexanal, and protein oxidation by carbonylation. Digests of n-3 PUFA-rich fish demonstrated the highest n-3 PUFA oxidation, whereas digests of various poultry and rabbit muscles showed highest n-6 PUFA oxidation, which correlated significantly with the n-6/n-3 PUFA ratio. Without lard addition, lipid oxidation is significantly higher in chicken and pork loin digests versus beef and deer digests, whereas the opposite occurred when these muscles are mixed with lard. Protein carbonylation correlates significantly with levels of TBARS and the sum of hydroxy-alkenals in digests. The n-6/n-3 PUFA ratio correlates well with the 4-hydroxy-2-nonenal/4-hydroxy-2-hexenal ratio in digests.. Muscular fatty acid profiles largely explain type and extent of lipid and protein oxidation during gastrointestinal digestion. Red meat only stimulates oxidation when digested with specific fat sources.

Topics: Aldehydes; Animals; Digestion; Fatty Acids, Omega-3; Fatty Acids, Omega-6; Fishes; Gastrointestinal Tract; Lipid Metabolism; Meat; Muscles; Oxidation-Reduction; Poultry

In this study, the formation of two toxic reactive aldehydes, 4-hydroxy-2-hexenal (HHE) and 4-hydroxy-2-nonenal (HNE), was investigated during frying of two different foodstuffs at 180 °C for 7 h in three different vegetable oils. The results showed that HHE and HNE content in the oil after frying was lower than that in the oil fried without foods. It was mainly because of the incorporation of HHE/HNE into the fried foods. In French Fries (FF), the HNE content was higher, whereas it was lower in the fried chicken breast meat (FCBM). The bidirectional model systems consisting of the model oil frying system and the model food frying system were used. The result of the model oil system showed that the content of HNE was higher in FF for the higher hydrophobic property than that in HHE, which would be preferably bounded into the hydrophobic helical structures, whereas the lower content of HNE was observed in FCBM due to its higher reactivity towards the nucleophilic group, namely, the protein in FCBM. Furthermore, the model food frying system including starch and protein extracted from the corresponding foodstuffs verified the results in the model oil system. Finally, the probable migration mechanism of HHE and HNE in different food matrices was proposed for the first time.

Topics: Aldehydes; Animals; Chickens; Cooking; Meat; Plant Oils; Solanum tuberosum

4-Hydroxy-2-alkenals disappear in the presence of food phenolics (i.e., cathechin or quercetin), and the corresponding carbonyl-phenol adducts are produced. In an attempt to identify structure(s) of formed adducts, the reactions between model phenolics (resorcinol, 2-methylresorcinol, orcinol, and 2,5-dimethylresorcinol) and hydroxyalkenals (4-hydroxy-2-hexenal and 4-hydroxy-2-nonenal) were studied and the produced adducts were isolated by column chromatography and unambiguously characterized by one- and two-dimensional nuclear magnetic resonance and mass spectrometry as dihydrobenzofuranols (1), chromane-2,7-diols (2), and 2 H-chromen-7-ols (3). These compounds were mainly produced at slightly basic pH values and moderate temperatures. Their activation energies ( E

Topics: Aldehydes; Catechin; Food; Hydrogen-Ion Concentration; Lipid Peroxidation; Magnetic Resonance Spectroscopy; Maillard Reaction; Phenol; Quercetin; Thermodynamics

The role of activation of lipid peroxidation in the mechanisms of acute methanol poisoning has not been studied.. We measured the concentrations of lipid peroxidation markers in acutely intoxicated patients with known serum concentrations of methanol and leukotrienes.. Blood serum samples were collected from 28 patients hospitalized with acute intoxication and from 36 survivors 2 years after discharge. In these samples, concentrations of 4-hydroxy-trans-2-hexenal (HHE), 4-hydroxynonenal (HNE), and malondialdehyde (MDA) were measured using the method of liquid chromatography-electrospray ionization-tandem mass spectrometry.. The maximum acute serum concentrations of all three lipid oxidative damage markers were higher than the follow-up serum concentrations: HNE 71.7 ± 8.0 ng/mL versus 35.4 ± 2.3 ng/mL; p < .001; HHE 40.1 ± 6.7 ng/mL versus 17.7 ± 4.1 ng/mL; p < .001; MDA 80.0 ± 7.2 ng/mL versus 40.9 ± 1.9 ng/mL; p < .001. The survivors without methanol poisoning sequelae demonstrated higher acute serum concentrations of the markers than the patients with sequelae. A correlation between measured markers and serum leukotrienes was present: HNE correlated with LTC4 (r = 0.663), LTD4 (r = 0.608), LTE4 (r = 0.771), LTB4 (r = 0.717), HHE correlated with LTC4 (r = 0.713), LTD4 (r = 0.676), LTE4 (r = 0.819), LTB4 (r = 0.746), MDA correlated with LTC4 (r = 0.785), LTD4 (r = 0.735), LTE4 (r = 0.814), LTB4 (r = 0.674); all p < .001. Lipid peroxidation markers correlated with anion gap (r= -0.428, -0.388, -0.334; p = .026, .045, .080 for HNE, HHE, MDA, respectively). The follow-up serum concentrations of lipid oxidation markers measured in survivors with and without visual/neurological sequelae 2 years after discharge did not differ.. Our results demonstrate that lipid peroxidation plays a significant role in the mechanisms of acute methanol poisoning. The acute concentrations of three measured biomarkers were elevated in comparison with the follow-up concentrations. Neuronal membrane lipid peroxidation seems to activate leukotriene-mediated inflammation as a part of the neuroprotective mechanisms. No cases of persistent elevation were registered among the survivors 2 years after discharge.

Topics: Activation, Metabolic; Alcoholism; Aldehydes; Biomarkers; Cysteine Proteinase Inhibitors; Female; Follow-Up Studies; Humans; Lipid Peroxidation; Male; Methanol; Middle Aged

In obesity-linked insulin resistance, oxidative stress in adipocytes leads to lipid peroxidation and subsequent carbonylation of proteins by diffusible lipid electrophiles. Reduction in oxidative stress attenuates protein carbonylation and insulin resistance, suggesting that lipid modification of proteins may play a role in metabolic disease, but the mechanisms remain incompletely understood. Herein, we show that

Topics: Adipose Tissue; Aldehydes; Amino Acid Sequence; Animals; Cell Nucleus; DNA-Binding Proteins; Mice; Nuclear Proteins; Obesity; Oxidative Stress; Protein Carbonylation; Zinc Fingers

This study investigated the effects of three light intensities on four types of palm oils during consecutive storage for 12 months at 4 °C. The concentrations of 4-hydroxy-2- trans-hexenal (4-HHE), 4-hydroxy-2- trans-nonenal (4-HNE), polycyclic aromatic hydrocarbon (PAH)4, and PAH8 in the oils significantly increased with the increasing light intensity after storage. The red palm oil had the lowest rate of increase of 4-HNE, while 5° palm oil had the highest rate of increase of the PAH, OPAH, 4-HNE, and peroxide values during storage. For the same type of oil, OPAHs increased significantly under a light intensity of 6000 lx (lx) after storage. The increasing concentrations of 9FO, ATQ, and BaPO in the oils stored at 6000 lx showed a positive relation to their corresponding parent PAHs, indicating that PAH oxidation occurred at 6000 lx. The results suggest that light intensity and β-carotene may control PAHs, OPAHs, and 4-hydroxy-trans- alkenals for vegetable oil storage, transportation, and retail.

Topics: Aldehydes; Antioxidants; beta Carotene; Fatty Acids, Unsaturated; Food Storage; Humans; Light; Oxidation-Reduction; Palm Oil; Peroxides; Polycyclic Aromatic Hydrocarbons; Temperature

A group of toxic aldehydes such as, malondialdehyde (MDA), 4-hydroxy-2-hexenal (HHE), and 4-hydroxy-2-nonenal (HNE) have been found in various vegetable oils and oil-based foods. Then simultaneous determination of them holds a great need in both the oil chemistry field and food field. In the present study, a simple and efficient analytical method was successfully developed for the simultaneous separation and detection of MDA, HHE, and HNE in vegetable oils by reversed-phase-high-performance liquid chromatography (RP-HPLC) coupled with photodiode array detector (PAD) at dual-channel detection mode. The effect of various experimental factors on the extraction performance, such as coextraction solvent system, butylated hydroxytoluene addition, and trichloroacetic acid addition were systematically investigated. Results showed that the linear ranges were 0.02-10.00 μg/mL for MDA, 0.02-4.00 μg/mL for HHE, and 0.03-4.00 μg/mL for HNE with the satisfactory correlation coefficient of >0.999 for all detected aldehydes. The limit of detection (LOD) and limit of quantification (LOQ) of MDA, HHE, and HNE were ∼0.021and 0.020 μg/mL, ∼0.009 and 0.020 μg/mL, and ∼0.014 and 0.030 μg/mL, respectively. Their recoveries were 99.64-102.18%, 102.34-104.61%, and 98.87-103.04% for rapeseed oil and 96.38-98.05%, 96.19-101.34%, and 96.86-99.04% for French fries, separately. Under the selected conditions, the developed methods was successfully applied to the simultaneous determination of MDA, HHE, and HNE in different tested vegetable oils. The results indicated that this method could be employed for the quality assessment of vegetable oils.

Topics: Aldehydes; Chromatography, High Pressure Liquid; Chromatography, Reverse-Phase; Malondialdehyde; Plant Oils

Marine lipids contain a high proportion of polyunsaturated fatty acids (PUFA), including the characteristic long chain (LC) n-3 PUFA. Upon peroxidation these lipids generate reactive products, such as malondialdehyde (MDA), 4-hydroxy-2-hexenal (HHE) and 4-hydroxy-2-nonenal (HNE), which can form covalent adducts with biomolecules and thus are regarded as genotoxic and cytotoxic. PUFA peroxidation can occur both before and after ingestion. The aim of this study was to determine what levels of MDA, HHE and HNE can evolve in the gastric and intestinal lumen after ingesting meals containing fish or fish oil using a dynamic gastrointestinal (GI) model (TIM). The impact of the fish muscle matrix, lipid content, fish species, and oven baking on GI oxidation was evaluated. MDA and HHE concentrations in gastric lumen increased for all meals during digestion, with the highest level found with herring mince; ∼ 25 μM MDA and ∼ 850 nM HHE. Aldehyde concentrations reached in intestinal lumen during digestion of fish containing meals were generally lower than in gastric lumen, while isolated herring oils (bulk and emulsified) generated higher MDA and HHE values in intestinal lumen compared to gastric lumen. Based on aldehyde levels in gastric lumen, meals containing herring lipids were ranked: raw herring (17% lipid) = baked herring (4% lipid) > raw herring (4% lipid) ≫ herring oil emulsion > herring oil. Herring developed higher concentrations of MDA and HHE during gastric digestion compared to salmon, which initially contained lower levels of oxidation products. Cooked salmon generated higher MDA concentrations during digestion than raw salmon. Low levels of HNE were observed during digestion of all test meals, in accordance with the low content of n-6 PUFA in fish lipids.

Topics: Aldehydes; Animals; Cooking; Digestion; Fatty Acids, Unsaturated; Fish Oils; Gastrointestinal Tract; Humans; Lipid Peroxidation; Malondialdehyde; Models, Biological; Oxidation-Reduction; Salmon; Seafood

In this work, we investigated lipid oxidation of cod liver oil during gastrointestinal (GI) digestion using two types of in vitro digestion models. In the first type of model, we used human GI juices, while we used digestive enzymes and bile from porcine origin in the second type of model. Human and porcine models were matched with respect to factors important for lipolysis, using a standardized digestion protocol. The digests were analysed for reactive oxidation products: malondialdehyde (MDA), 4-hydroxy-trans-2-nonenal (HNE), and 4-hydroxy-trans-2-hexenal (HHE) by liquid chromatography/atmospheric pressure chemical ionization-mass spectrometry (LC/APCI-MS), and for free fatty acids (FFA) obtained during the digestion by gas chromatography-mass spectrometry (GC-MS). The formation of the oxidation products MDA, HHE, and HNE was low during the gastric digestion, however, it increased during the duodenal digestion. The formation of the oxidation products reached higher levels when digestive juices of human origin were used (60 μM of MDA, 9.8 μM of HHE, and 0.36 μM of HNE) [corrected] compared to when using enzymes and bile of porcine origin (0.96, and 1.6 μM of MDA; 0.16, and 0.23 μM of HHE; 0.026, [corrected] and 0.005 μM of HNE, respectively, in porcine models I and II). In all models, FFA release was only detected during the intestinal step, and reached up to 31% of total fatty acids (FA). The findings in this work may be of importance when designing oxidation oriented lipid digestion studies.

Topics: Aldehydes; Animals; Cod Liver Oil; Digestion; Gastrointestinal Tract; Humans; Malondialdehyde; Oxidation-Reduction; Swine

Marine long-chain polyunsaturated fatty acids (LC n-3 PUFA) are associated with reduced risk for inflammatory diseases, such as cardiovascular diseases and rheumatoid arthritis. These fatty acids, however, are rapidly oxidized, generating highly reactive malondialdehyde (MDA), 4-hydroxy-2-hexenal (HHE) and 4-hydroxy-2-nonenal (HNE). These oxidation products may interact with DNA and proteins, thus possibly leading to impaired cell functions. Little is known about the formation of MDA, HHE and HNE in fish oil in the gastrointestinal (GI) tract. In this study, the effect of dynamic in vitro digestion of cod liver oil on the generation of MDA, HHE and HNE was evaluated using the TNO Gastro-Intestinal Model (tiny-TIM). Effects of pre-formed oxidation products, pre-emulsification of the oil, and addition of oxidants (EDTA and hemoglobin, Hb) on GI oxidation were evaluated. Formation of aldehydes occurred during GI digestion. However, only emulsified oil fortified with 11.5 μM Hb oxidized to a degree that overcame the dilution induced by gastric secretion, which caused increased aldehyde concentrations in gastric lumen up to 90 min. The maximum levels of aldehydes generated in this study were 24.5 μM MDA, 1.6 μM HHE and 0.07 μM HNE. Oils containing different amounts of pre-formed lipid oxidation products maintained the same oxidation ranking order during digestion, even though the relative changes were not directly proportional. Emulsification of the oil had an unclear effect in the gastric phase, but a pro-oxidative effect in the intestinal phase. In general, higher aldehyde levels were reached in the intestinal lumen than in the initial meal, demonstrating that GI digestion promotes oxidation. Hence, epithelial cells may be exposed to elevated amounts of reactive aldehydes for several hours after a meal containing fish oil.

Topics: Aldehydes; Ascorbic Acid; Cod Liver Oil; Digestion; Edetic Acid; Epithelial Cells; Gastrointestinal Tract; Hemoglobins; Humans; Malondialdehyde; Models, Biological

Membrane transporters are involved in enormous number of physiological and pathological processes. Under oxidative stress they become targets for reactive oxygen species and its derivatives which cause protein damage and/or influence protein function(s). The molecular mechanisms of this interaction are poorly understood. Here we describe a novel lipid-mediated mechanism by which biologically important reactive aldehydes (RAs; 4-hydroxy-2-nonenal, 4-hydroxy-2-hexenal and 4-oxo-2-nonenal) modify the activity of several membrane transporters. We revealed that investigated RAs covalently modify the membrane lipid phosphatidylethanolamine (PE), that lead to the formation of different membrane active adducts. Molecular dynamic simulations suggested that anchoring of PE-RA adducts in the lipid headgroup region is primarily responsible for changes in the lipid membrane properties, such as membrane order parameter, boundary potential and membrane curvature. These caused the alteration of transport activity of mitochondrial uncoupling protein 1, potassium carrier valinomycin and ionophore CCCP. In contrast, neither direct protein modification by RAs as previously shown for cytosolic proteins, nor its insertion into membrane bilayers influenced the studied transporters. Our results explain the diversity of aldehyde action on cell proteins and open a new field in the investigation of lipid-mediated effects of biologically important RAs on membrane receptors, channels and transporters.

Topics: Aldehydes; Fatty Acids; Humans; Ion Channels; Lipid Bilayers; Mitochondrial Proteins; Molecular Dynamics Simulation; Potassium; Protein Conformation; Protons; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Uncoupling Protein 1

Malondialdehyde (MDA), 4-Hydroxy-2-(E)-Nonenal (HNE) and 4-Hydroxy-2-(E)-Hexenal (HHE) are reactive aldehydes found in foods and are formed due to decomposition of polyunsaturated fatty acid hydroperoxides. In the present study, sixteen food categories were analyzed for the aforementioned aldehydes and in combination with consumption data obtained from a national representative sample of the Belgian population, a quantitative exposure assessment was performed. MDA was detected above the detection limit in 84% of the analyzed samples while HNE and HHE in 63% and 16% of the samples respectively. Consumption of dry nuts, fried snacks, French fries and cured minced meat products were found to contribute the most to the intake of MDA and HNE. Intake of HHE from the foods analyzed was found not to be significant. An evaluation of any potential risk related to the intake of the studied aldehydes through the studied foods was performed by applying the threshold of toxicological concern concept. No risk to human health could be identified related to the consumption of these foods for the vast majority of the consumers, with the only exception of a small proportion (3.8%) of those who consume cured and minced raw meat, that could be at risk.

Topics: Aldehydes; Belgium; Diet; Environmental Exposure; Food Contamination; Humans; Malondialdehyde; Risk Assessment

We report a mass spectrometry-based comparative "bottom up" proteomics approach that combines d(0)/d(4)-succinic anhydride labeling with commercially available hydrazine (Hz)-functionalized beads (Affi-gel Hz beads) for detection, identification and relative quantification of site-specific oxylipid modifications in biological matrices. We evaluated and applied this robust and simple method for the quantitative analysis of oxylipid protein conjugates in cardiac mitochondrial proteome samples isolated from 3- and 24-month-old rat hearts. The use of d(0)/d(4)-succinic anhydride labeling, Hz-bead based affinity enrichment, nanoLC fractionation and MALDI-ToF/ToF tandem mass spectrometry yielded relative quantification of oxylipid conjugates with residue-specific modification information. Conjugation of acrolein (ACR), 4-hydroxy-2-hexenal (HHE), 4-hydroxy-2-nonenal (HNE) and 4-oxo-2-noneal (ONE) to cysteine, histidine and lysine residues were identified. HHE conjugates were the predominant subset of Michael-type adducts detected in this study. The HHE conjugates showed higher levels in mitochondrial preparations from young heart congruent with previous findings by others that the n-3/n-6 PUFA ratio is higher in young heart mitochondrial membranes. Although this study focuses on protein adducts of reactive oxylipids, the method might be equally applicable to protein carbonyl modifications caused by metal catalyzed oxidation reactions.

Topics: Aging; Aldehydes; Animals; Deuterium; Hydrazines; Lipid Peroxidation; Lipids; Male; Mitochondria, Heart; Protein Processing, Post-Translational; Proteomics; Rats; Rats, Inbred F344; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Succinic Anhydrides

Lipid peroxidation produces many reactive byproducts including 4-hydroxy-2-hexenal (HHE) and 4-hydroxy-2-nonenal (HNE) derived from the peroxidation of n-3 and n-6 polyunsaturated fatty acids, respectively. HNE and HHE can modify circulating biomolecules through the formation of covalent adducts. It remains, however, unknown whether HHE and HNE could induce functional and structural changes in the insulin molecule, which may in turn be pivotal in the development of insulin resistance and diabetes. Recombinant human insulin was incubated in the presence of HHE or HNE, and the formation of covalent adducts on insulin was analyzed by mass spectrometry analysis. Insulin tolerance test in mice and stimulation of glucose uptake by 3T3 adipocytes and L6 muscle cells were used to evaluate the biological efficiency of adducted insulin compared with the native one. One to 5 adducts were formed on insulin through Michael adduction, involving histidine residues. Glucose uptake in 3T3-L1 and L6C5 cells as well as the hypoglycemic effect in mice was significantly reduced after treatment with adducted insulin compared to native insulin. The formation of HNE- and HHE-Michael adducts significantly disrupts the biological activity of insulin. These structural and functional abnormalities of the insulin molecule might contribute to the pathogenesis of insulin resistance.

Topics: 3T3-L1 Cells; Aldehydes; Amino Acid Sequence; Animals; Cell Line; Humans; Insulin; Lipid Peroxidation; Mice; Models, Molecular; Molecular Sequence Data; Oxidative Stress; Rats; Recombinant Proteins

Ethanol withdrawal increases lipid peroxidation of the polyunsaturated fatty acid (PUFA) docosahexaenoate (22:6; n-3) in the CNS. To further define the role of oxidative damage of PUFAs during ethanol withdrawal, we measured the levels of glutathione adducts of 4-hydroxy-2-hexenal (GSHHE) and 4-hydroxy-2-nonenal (GSHNE) as biomarkers of brain lipid peroxidation of n-3 and n-6 PUFAs, respectively. In this study rats received an ethanol-containing diet for 6 weeks followed by withdrawal ranging from 0 to 7 days. GSHHE content was elevated (>350%) in the cerebral cortex after 2 days of withdrawal with no change in GSHNE. The levels of GSHHE were significantly greater (2- to 20-fold) than those of GSHNE in multiple brain regions. Experiments demonstrated that intoxication and withdrawal did not alter the enzymatic rate of formation of GSHHE or GSHNE, but the rate of formation of GSHHE was higher (approximately 50%) than that of GSHNE. These results indicate that selective oxidative damage to n-3 PUFAs occurs in the cerebral cortex as a result of ethanol withdrawal and that 4-hydroxy-2-hexenal is metabolized to the GSH adduct more efficiently than HNE.

Topics: Aldehydes; Animals; Brain; Cerebral Cortex; Chromatography, Liquid; Ethanol; Fatty Acids; Glutathione; Lipid Peroxidation; Male; Rats; Rats, Sprague-Dawley; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Since damage to DNA and other cellular molecules by reactive oxygen species ranks high as a major culprit in the onset and development of colorectal cancer, the aim of the present study is to clarify the role of antioxidant seleonoproteins including glutathione peroxidase (GPx), thioredoxin reductase (TXR) and selenoprotein P (SePP), and the effect of oxidative stress on the progression of colorectal cancer. Expression of 14 oxidative stress-related molecules in both tumorous and non-tumorous tissues in 41 patients was examined by immunohistochemistry and Western blot analysis. Expression levels of proteins modified by 4-hydroxy-2-nonenal (4-HNE), malonyldialdehyde (MDA) and 4-hydroxy-2-hexenal (4-HHE), and the positive rate of 8-hydroxy-2'-deoxyguanosine (8-OHdG) in tumorous tissues were much higher than those in non-tumorous tissues. Glutathione (GSH) content in tumor tissues was much lower than that in non-tumorous tissues. Expression level of selenoproteins such as GPx-1, GPx-3, and SePP, which are rapidly degraded during selenium deprivation, was significantly decreased in tumorous tissues, whereas that of GPx-2, which is resistant to selenium deprivation, was increased. Expression of SePP was decreased at stage III and IV, compared to that of stage II. These data suggest that contrasting expression pattern of the antioxidant selenoproteins plays an important role in the progression of colorectal cancer.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Aged; Aldehydes; Antioxidants; Apoptosis; Blotting, Western; Cell Proliferation; Colorectal Neoplasms; Deoxyguanosine; Disease Progression; Female; Glutathione; Glutathione Peroxidase; Humans; Immunohistochemistry; Male; Malondialdehyde; Oxidative Stress; Proliferating Cell Nuclear Antigen; Selenoprotein P; Selenoproteins; Superoxide Dismutase; Superoxide Dismutase-1; Thioredoxin-Disulfide Reductase; Tumor Suppressor Protein p53

Lipid peroxidation of docosahexaenoic (22:6; n-3) acid (DHA) is elevated in the CNS in patients with Alzheimer's disease and in animal models of seizure and ethanol withdrawal. One product of DHA oxidation is trans-4-hydroxy-2-hexenal (HHE), a six carbon analog of the n-6 fatty acid derived trans-4-hydroxy-2-nonenal (HNE). In this work, we studied the neurotoxic potential of HHE. HHE and HNE were toxic to primary cultures of cerebral cortical neurons with LD(50)'s of 23 and 18 micromol/L, respectively. Toxicity was prevented by the addition of thiol scavengers. HHE and HNE depleted neuronal GSH content identically with depletion observed with 10 micromol/L of either compound. Using an antibody raised against HHE-protein adducts, we show that HHE modified specific proteins of 75, 50, and 45 kDa in concentration- and time-dependent manners. The time-dependent formation of HHE differed from that of F4-neuroprostanes following in vitro DHA oxidation likely as a result of the different oxidation pathways involved. Using purified mitochondrial aldehyde dehydrogenase ALDH5A, we found that HHE was oxidized 6.5-fold less efficiently than HNE. Our data demonstrate that HHE and HNE have similarities but also differences in their neurotoxic mechanisms and metabolism.

Topics: Aldehydes; Animals; Brain Injury, Chronic; Cells, Cultured; Cerebral Cortex; Docosahexaenoic Acids; Dose-Response Relationship, Drug; Female; Free Radical Scavengers; Glutathione; Lipid Peroxidation; Nerve Tissue Proteins; Neurodegenerative Diseases; Neurons; Neurotoxins; Oxidative Stress; Rats; Succinate-Semialdehyde Dehydrogenase; Time Factors

Protein modifications resulting from reactive aldehydes are thought to be involved in the pathogenesis of various degenerative diseases. Aged cynomolgus monkey (Macaca fascicularis) spontaneously develop drusen in the macula, consistent with the phenotype observed in early-stage age-related macular degeneration (AMD), indicating that this animal is an optimum model for AMD. In retinal sections from three monkeys with macular degeneration, regardless of their size, drusen were consistently positive with immunohistochemical labeling against protein modifications by 4-hydroxynonenal and 4-hydroxyhexenal, end products of non-enzymatic oxidation of n-6 and n-3 polyunsaturated fatty acids, respectively. Positive labeling for both modifications was observed in the ganglion cell layer, the inner nuclear layer, the outer nuclear layer, and the retinal pigment epithelium. However, no consistent differences in location or intensity of the labeling were observed between monkeys with normal macula and macular degeneration. The results suggest a possible association between drusen formation and protein modifications by aldehydes in the pathogenesis of AMD.

Topics: Aldehydes; Animals; Disease Models, Animal; Eye Proteins; Female; Macaca fascicularis; Macular Degeneration; Retinal Drusen

An isotopically coded affinity probe was developed and evaluated for the characterization and quantification of proteins adducted by 2-alkenals derived from lipid peroxidation (LPO) processes. Lipid-derived 2-alkenals, such as acrolein and 4-hydroxy-2-nonenal (HNE), have the ability to react with cysteine, histidine, and lysine residues in proteins, thus causing protein damage and loss of protein function. Such modifications of proteins are difficult to characterize in biological samples by mass spectrometry due to the complexity of protein extracts and the low abundance of adducted proteins. The novel aldehyde-reactive, hydrazide-functionalized, isotope-coded affinity tag (HICAT) described in this study was found effective for the selective isolation, detection, and quantification of Michael-type adducts of 2-alkenals with proteins using a combination of affinity isolation, nanoLC, and matrix-assisted laser desorption ionization tandem mass spectrometry (MALDI-MS/MS). The chemical and mass spectrometric properties of the new probe are demonstrated on a model protein treated with HNE. The efficacy of HICAT for the analysis of complex samples was tested using preparations of mitochondrial proteins that were modified in vitro with HNE. The potential of the HICAT strategy for the identification, characterization, and quantification of in vivo oxylipid-protein conjugates is demonstrated on cardiac mitochondrial protein preparations, in which, for example, the ADP/ATP translocase 1 was found adducted to the 2-alkenals, acrolein and 4-hydroxy-2-hexenal, at Cys-256.

Topics: Acrolein; Aldehydes; Alkenes; Animals; Carbon Isotopes; Chromatography, High Pressure Liquid; Hydrazines; Lipid Peroxidation; Lipids; Mitochondria, Heart; Mitochondrial Proteins; Molecular Probes; Molecular Structure; Rats; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

4-Hydroxy-2-hexenal (HHE) and 4-hydroxy-2-nonenal (HNE) were determined using selected ion-monitoring gas chromatography-mass spectrometry (GC-MS) in 56 kinds of commercially available PUFA-fortified foods including infant formulas and baby foods. HHE and HNE, each specifically coming from the oxidation of n-3 and n-6 polyunsaturated fatty acids (PUFA), were observed at <10-77 and 41-132 microg kg(-1) in the infant formulas (n = 12) and at <10-52 and 36-116 microg kg(-1) in the baby foods (n = 7), respectively. 4-Hydroxy-2-alkenals in infant formulas and baby foods were further determined at 10 and 30 days after opening in an attempt to examine the time dependence of the levels of 4-hydroxy-2-alkenals. The values of HHE and HNE had increased appreciably to <10-220 and 79-792 microg kg(-1) in infant formulas and to <10-112 and 135-572 microg kg(-1) in baby foods, respectively, at 10 days and decreased, although statistically not significant, in most of the tested samples after 30 days, which suggested that the reactive compounds might interact with other constituents like proteins in the samples to form adducts or be decomposed with time. Based on the current study, it was calculated that 3-month to 1-year-old babies maintained exclusively on these commercially available PUFA-fortified infant formulas or baby foods could be exposed to a maximum of 20.2 microg kg(-1) body weight day(-1) of 4-hydroxy-2-alkenals, which is two orders of magnitude higher than the exposure of Korean adults estimated in a previous study of the authors' (2005). The present study may trigger future studies investigating the physiological influence of 4-hydroxy-2-alkenals originating from the diet on man at an early stage of development.

Topics: Aldehydes; Fatty Acids, Unsaturated; Food Analysis; Food Contamination; Food, Fortified; Gas Chromatography-Mass Spectrometry; Humans; Infant; Infant Food; Infant Formula; Lipid Peroxidation

Oxidative stress plays a role in pathogenesis of chronic viral hepatitis. Expression of oxidative stress-related molecules remains to be clarified.. 4-hydroxy-2-nonenal (4-HNE), 4-hydroxy-2-hexenal (4-HHE), catalase, superoxide dismutase-1 (SOD-1), glutathione peroxidase-1, thioredoxin (TRX) in leukocytes and urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) were examined in 164 persons, including 130 chronic viral hepatitis patients and 34 normal individuals, by Western blot analysis and enzyme-linked immunosorbent assay, respectively. Hepatic expression of these proteins was immunohistochemically examined in 12 patients with chronic viral hepatitis, compared with three persons without liver damage.. The 4-HNE/beta-actin ratios in chronic viral hepatitis were significantly higher than those in normal individuals (P<0.01), and were significantly correlated with asparate aminotransferase (AST) and alanine aminotransferase (ALT) (P<0.01, each). The catalase/beta-actin and SOD-1/beta-actin ratios in chronic viral hepatitis were higher than those in normal individuals, and were significantly correlated with 4-HNE/beta-actin ratios (P<0.01, each). Hepatic expression of 4-HNE, 4-HHE, catalase, SOD-1 and TRX in chronic viral hepatitis was higher than that without liver damage. Urinary excretion of 8-OHdG was not changed in chronic viral hepatitis.. The results of the present study suggest that expression of oxidative stress-related molecules in leukocytes is upregulated in relation to serum aminotransferase levels.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Aged; Alanine Transaminase; Aldehydes; Aspartate Aminotransferases; Blotting, Western; Deoxyguanosine; Enzyme-Linked Immunosorbent Assay; Enzymes; Female; Hepatitis, Viral, Human; Humans; Leukocytes; Lipid Peroxidation; Liver; Male; Middle Aged; Oxidative Stress; Thioredoxins; Up-Regulation

Lipid peroxidation products such as 4-hydroxy-2-nonenal (HNE) may be responsible for various pathophysiological events under oxidative stress, since they injure cellular components such as proteins and DNA. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which is a key enzyme of glycolysis and has been reported to be a multifunctional enzyme, is one of the enzymes inhibited by HNE. Previous studies showed that GAPDH is degraded when incubated with acetylleucine chloromethyl ketone (ALCK), resulting in the liberation of a 23-kDa fragment. In this study, we examined whether GAPDH incubated with HNE or other aldehydes of lipid peroxidation products are degraded similarly to that with ALCK. The U937 cell extract was incubated with these aldehydes at 37 degrees C and analyzed by Western blotting using anti-GAPDH antibodies. Incubation with HNE or 4-hydroxy-2-hexenal (HHE) decreased GAPDH activity and GAPDH protein level, and increased the 23-kDa fragment, in time- and dose-dependent manners, but that with other aldehydes did not. Gel filtration using the Superose 6 showed that the GAPDH-degrading activity was eluted in higher molecular fractions than proteasome activity. The enzyme activity was detected at the basic range of pH and inhibited by serine protease inhibitors, diisopropyl fluorophosphate and phenylmethylsulfonyl fluoride, but not by other protease inhibitors including a proteasome inhibitor, MG-132, and a tripeptidyl peptidase II (TPP II) inhibitor, AAF-CMK. These results suggest that GAPDH modified by HNE and HHE is degraded by a giant serine protease, releasing the 23-kDa fragment, not by proteasome or TPP II.

Topics: Aldehydes; Amino Acid Chloromethyl Ketones; Blotting, Western; Chromatography, Gel; Enzyme Inhibitors; Glyceraldehyde-3-Phosphate Dehydrogenases; Glycolysis; Humans; Hydrogen-Ion Concentration; Inhibitory Concentration 50; Leupeptins; Lipid Peroxidation; Oxidative Stress; Phenylmethylsulfonyl Fluoride; Proteasome Endopeptidase Complex; Sepharose; Serine Endopeptidases; Time Factors; U937 Cells

Oxidative stress contributes to the pathogenesis of chronic hepatitis C. The aim of this study was to assess the peroxidation of n-3 polyunsaturated fatty acids (PUFAs) in the liver and its relation to hepatic steatosis in chronic hepatitis C.. We immunohistochemically detected malondialdehyde (MDA)-, 4-hydroxy-2-nonenal (HNE)-, and 4-hydroxy-2-hexenal (HHE)-protein adducts in liver biopsy specimens from 55 patients with chronic hepatitis C. Cells stained positively for HHE-protein adducts were quantified using computer-based image analysis. Fatty-acid composition was determined, by gas chromatography, for the noncancerous portions of resected livers, with or without steatosis, obtained from two patients with hepatitis C virus-associated hepatocellular carcinoma.. The detection rate of HHE-protein adducts (63.6%) was significantly higher than that of MDA-protein adducts (21.8%; P < 0.001) or HNE-protein adducts (29.1%; P < 0.001). Areas positively stained for HHE-protein adducts (HHE-positive areas) were significantly larger in 18 patients with steatosis (6.2 +/- 3.6%) than in 17 patients without steatosis (3.4 +/- 2.6%; P = 0.01). Resected liver tissue with steatosis showed a larger HHE-positive area (18.6%) and higher ratio of n-6 PUFA content to n-3 PUFA content (3 : 1) than liver tissue without steatosis (7.2%; 2 : 3). On multivariate analysis, the HHE-positive area (odds ratio, 1.55; 95% confidence interval [CI], 1.08-2.23; P = 0.019) was a factor associated with the presence of hepatic steatosis.. HHE-protein adducts, which are a good marker for oxidative stress, are associated with steatosis in chronic hepatitis C.

Topics: Adult; Aged; Aged, 80 and over; Aldehydes; Biomarkers; Biopsy; Chromatography, Gas; Fatty Acids, Omega-3; Fatty Acids, Unsaturated; Fatty Liver; Female; Hepatitis C, Chronic; Humans; Immunohistochemistry; Lipid Peroxidation; Male; Malondialdehyde; Middle Aged; Oxidative Stress; Severity of Illness Index; Triglycerides

4-Hydroxy-2-alkenals are cytotoxic aldehydes generated from the oxidation of n-3 and n-6 polyunsaturated fatty acids. The compounds have shown various biological effects via Schiff base adducts or Michael addition adducts at levels higher than physiological ones. To assess human exposure to 4-hydroxy-2-alkenals in the diet, 4-hydroxy-2-alkenals in vegetable oils, fish and shellfish were monitored using GC/MS/SIM. 2001 National Health and Nutrition Survey (2002) data were employed for the dietary intake pattern. The Korean daily exposure to 4-hydroxy-2-alkenals, excluding a possible one from fried food, was 4.3 microg day-1, constituted of 1.6 microg 4-hydroxy-2-hexenal (HHE) and 2.7 microg 4-hydroxy-2-nonenal (HNE). It was calculated that Koreans could be additionally exposed to more than 11.8 microg day-1 4-hydroxy-2-alkenal from fried foods. Thus, the combined exposure would be 16.1 microg day-1, which corresponds to 0.3 microg kg-1 body weight day-1 for a 60 kg Korean adult. In spite of the biological toxicity of 4-hydroxy-2-alkenals, the risk for human could not be quantified due to the lack of a virtually safe dose of the compounds. However, considering the basal level of 4-hydroxy-2-alkenals in many tissues, the present value from the diet may not pose a significant risk for human health.

Topics: Adult; Aldehydes; Animals; Fatty Acids, Unsaturated; Fishes; Food Analysis; Humans; Korea; Nutrition Surveys; Plant Oils; Risk Assessment; Shellfish

4-Hydroxynonenal (4-HNE) and 4-hydroxyhexenal (4-HHE) are reactive aldehydes derived from the nonenzymatic oxidation of n-6 and n-3 polyunsaturated fatty acids, respectively. Increasing evidence suggests that protein modifications by reactive aldehydes are involved in various diseases. The present study was undertaken to test whether protein modifications by 4-HNE and 4-HHE increase in retinal tissues after exposure of rats to damaging levels of light.. Albino rats were exposed to 1 or 5 klux white fluorescent light for 3 hours and, at various times thereafter, the levels and localizations of aldehyde-modified proteins in retinas were assessed by densitometric analysis of semiquantitative Western dot blots and by immunohistochemistry, using 4-HNE- and 4-HHE-specific antibodies. In some rats, the protective antioxidant phenyl-N-tert-butylnitrone (PBN) was injected (50 mg/kg) before exposure to light. To assess retinal damage, outer nuclear layer (ONL) thickness was measured on hematoxylin-eosin (H&E)-stained sections, and apoptosis was semiquantitatively analyzed by TUNEL staining.. By dot blot analysis, 4-HNE- and 4-HHE-modified proteins were significantly increased in retina (both by 1.7-fold) and RPE fraction (1.5- and 1.8-fold, respectively) after 5-klux exposure. In retina, increases in 4-HNE- and 4-HHE-modified proteins were more prominent at 3 hours than at 24 hours or 48 hours after exposure to light. In rod outer segments, only 4-HHE-modified proteins increased significantly (1.4-fold). Retinal thinning, TUNEL staining in ONL, 4-HNE-, and 4-HHE protein modifications were all found in the same retinal regions. PBN treatment inhibited the light-induced increase of 4-HNE and 4-HHE modified proteins in retina and RPE fractions.. Exposure to intense light increases 4-HNE and 4-HHE protein modifications in the retina, suggesting that free radical initiated, nonenzymatic reactions are involved in this process. These modifications may be early events that precede photoreceptor cell apoptosis.

Topics: Aldehydes; Animals; Apoptosis; Blotting, Western; Cyclic N-Oxides; Eye Proteins; Female; Free Radical Scavengers; Immunoenzyme Techniques; In Situ Nick-End Labeling; Light; Nitrogen Oxides; Protein Processing, Post-Translational; Radiation Injuries, Experimental; Rats; Rats, Sprague-Dawley; Retina; Retinal Degeneration

Lipid oxidation is implicated in a wide range of pathophysiogical disorders, and leads to reactive compounds such as fatty aldehydes, of which the most well known is 4-hydroxy-2E-nonenal (4-HNE) issued from 15-hydroperoxyeicosatetraenoic acid (15-HpETE), an arachidonic acid (AA) product. In addition to 15-HpETE, 12(S)-HpETE is synthesized by 12-lipoxygenation of platelet AA. We first show that 12-HpETE can be degraded in vitro into 4-hydroxydodeca-(2E,6Z)-dienal (4-HDDE), a specific aldehyde homologous to 4-HNE. Moreover, 4-HDDE can be detected in human plasma. Second, we compare the ability of 4-HNE, 4-HDDE, and 4-hydroxy-2E-hexenal (4-HHE) from n-3 fatty acids to covalently modify different ethanolamine phospholipids (PEs) chosen for their biological relevance, namely AA- (20: 4n-6) or docosahexaenoic acid- (22:6n-3) containing diacyl-glycerophosphoethanolamine (diacyl-GPE) and alkenylacyl-glycerophosphoethanolamine (alkenylacyl-GPE) molecular species. The most hydrophobic aldehyde used, 4-HDDE, generates more adducts with the PE subclasses than does 4-HNE, which itself appears more reactive than 4-HHE. Moreover, the aldehydes show higher reactivity toward alkenylacyl-GPE compared with diacyl-GPE, because the docosahexaenoyl-containing species are more reactive than those containing arachidonoyl. We conclude that the different PE species are differently targeted by fatty aldehydes: the higher their hydrophobicity, the higher the amount of adducts made. In addition to their antioxidant potential, alkenylacyl-GPEs may efficiently scavenge fatty aldehydes.

Topics: Aldehydes; Animals; Binding Sites; Brain; Chromatography, High Pressure Liquid; Eicosapentaenoic Acid; Gas Chromatography-Mass Spectrometry; Humans; Phosphatidylethanolamines; Phospholipids; Rats; Time Factors

Elevated levels of 4-hydroxy-trans-2-nonenal (HNE) are implicated in the pathogenesis of numerous neurodegenerative disorders. Although well-characterized in the periphery, the mechanisms of detoxification of HNE in the CNS are unclear. HNE is oxidized to a non-toxic metabolite in the rat cerebral cortex by mitochondrial aldehyde dehydrogenases (ALDHs). Two possible ALDH enzymes which might oxidize HNE in CNS mitochondria are ALDH2 and succinic semialdehyde dehydrogenase (SSADH/ALDH5A). It was previously established that hepatic ALDH2 can oxidize HNE. In this work, we tested the hypothesis that SSADH oxidizes HNE. SSADH is critical in the detoxification of the GABA metabolite, succinic semialdehyde (SSA). Recombinant rat SSADH oxidized HNE and other alpha,beta-unsaturated aldehydes. Inhibition and competition studies in rat brain mitochondria showed that SSADH was the predominant oxidizing enzyme for HNE but only contributed a portion of the total oxidizing activity in liver mitochondria. In vivo administration of diethyldithiocarbamate (DEDC) effectively inhibited (86%) ALDH2 activity but not HNE oxidation in liver mitochondria. The data suggest that a relationship between the detoxification of SSA and the neurotoxic aldehyde HNE exists in the CNS. Furthermore, these studies show that multiple hepatic aldehyde dehydrogenases are able to oxidize HNE.

Topics: Aldehyde Dehydrogenase; Aldehyde Dehydrogenase, Mitochondrial; Aldehyde Oxidoreductases; Aldehydes; Animals; Benomyl; Brain Chemistry; Ditiocarb; Enzyme Inhibitors; gamma-Aminobutyric Acid; Male; Mitochondria; Mitochondria, Liver; Oxidation-Reduction; Rats; Rats, Sprague-Dawley; Succinate-Semialdehyde Dehydrogenase

P1-zeta-crystallin (P1-ZCr) is an oxidative stress-induced NADPH:quinone oxidoreductase in Arabidopsis thaliana, but its physiological electron acceptors have not been identified. We found that recombinant P1-ZCr catalyzed the reduction of 2-alkenals of carbon chain C(3)-C(9) with NADPH. Among these 2-alkenals, the highest specificity was observed for 4-hydroxy-(2E)-nonenal (HNE), one of the major toxic products generated from lipid peroxides. (3Z)-Hexenal and aldehydes without alpha,beta-unsaturated bonds did not serve as electron acceptors. In the 2-alkenal molecules, P1-ZCr catalyzed the hydrogenation of alpha,beta-unsaturated bonds, but not the reduction of the aldehyde moiety, to produce saturated aldehydes, as determined by gas chromatography/mass spectrometry. We propose the enzyme name NADPH:2-alkenal alpha,beta-hydrogenase (ALH). A major portion of the NADPH-dependent HNE-reducing activity in A. thaliana leaves was inhibited by the specific antiserum against P1-ZCr, indicating that the endogenous P1-ZCr protein has ALH activity. Because expression of the P1-ZCr gene in A. thaliana is induced by oxidative stress treatments, we conclude that P1-ZCr functions as a defense against oxidative stress by scavenging the highly toxic, lipid peroxide-derived alpha,beta-unsaturated aldehydes.

Topics: Alcohol Oxidoreductases; Aldehyde Reductase; Aldehydes; Aldo-Keto Reductases; Alkenes; Arabidopsis; Catalysis; Electron Transport; Gas Chromatography-Mass Spectrometry; Glutathione Transferase; Hydrogen-Ion Concentration; Hydrogenation; Lipid Peroxidation; Models, Chemical; NAD(P)H Dehydrogenase (Quinone); Recombinant Proteins; Substrate Specificity; zeta-Crystallins

A facile one-pot synthesis of (E)-4-hydroxy-2-alkenals such as (E)-4-hydroxy-2-nonenal, (E)-4-hydroxy-2-heptenal, and (E)-4-hydroxy-2-hexenal was achieved from the corresponding (2E,4E)-2,4-alkadienals by reduction-oxygenation with molecular oxygen and triethylsilane in the presence of cobalt(II) porphyrin as a catalyst followed by treatment with trimethylphosphite.

Topics: Aldehydes; Cobalt; Fatty Alcohols; Magnetic Resonance Spectroscopy; Mass Spectrometry; Metalloporphyrins; Organometallic Compounds; Oxidation-Reduction

The effects of some 4-hydroxyalkenals, carbonylic products of lipid peroxidation, on hepatic phosphatidylinositol-4,5-bisphosphate (PIP2)-phospholipase C (PL-C) activity were investigated. The enzymatic activity was assayed in vitro by measuring the hydrolysis of [3H]PIP2 added as exogenous substrate to liver membranes. 4-Hydroxyhexenal (HEE), 4-hydroxyoctenal (HOE) and 4-hydroxynonenal (HNE) were able to stimulate both the basal and the GTPgammaS induced PL-C activity, whereas 4-hydroxyundecenal was inactive. HOE was the most active compound, being able to accelerate PIP2 breakdown at concentrations between 10(-12) and 10(-6) M, while in the case of HEE the effective doses ranged from 10(-11) to 10(-7) M and from 10(-9) to 10(-6) M in the case of HNE. 4-Hydroxynonenal was able to increase also bombesin stimulated PL-C activity. As these aldehydes accelerated PIP2 breakdown at doses which can be actually reached in tissues, the effects shown in vitro are likely to occur in vivo.

Topics: Aldehydes; Animals; Glutathione; Lipid Peroxidation; Liver; Male; Phosphatidylinositol 4,5-Diphosphate; Phosphatidylinositols; Phosphoinositide Phospholipase C; Phosphoric Diester Hydrolases; Rats; Rats, Inbred Strains; Type C Phospholipases

Under in vitro conditions, trans-4-hydroxy-2-hexenal (t-4HH), trans-4-hydroxy-2-nonenal (t-4-HN) and trans-2-hexenal (t-2H) significantly reduced the levels of mouse liver microsomal cytochrome P-450. Incubation of trans-4-hydroxy-alkenals, under anaerobic conditions in the absence of an NADPH-generating system indicated that these compounds were converting cytochrome P-450 to cytochrome P-420. Prior activation by the mixed function oxidase system was not required for trans-4-hydroxy-alkenals to alter cytochrome P-450 concentrations. trans-4-Hydroxy-alkenals and non-hydroxylated alpha,beta-unsaturated aldehydes may be exerting their effects on cytochrome P-450 by binding to sulfhydryl groups in a similar manner as reported for sulfhydryl reagents such as p-chloromercuriphenylsulfonic acid and p-chloromercuribenzoate.

Topics: Aldehydes; Animals; Cytochrome P-450 Enzyme System; Cytochromes; Male; Mice; Mice, Inbred BALB C; Microsomes, Liver; NADP; Structure-Activity Relationship

trans-4-Hydroxy-2-hexenal (t-4HH), a reactive metabolite isolated from the pyrrolizidine alkaloid senecionine, and trans-4-hydroxy-2-nonenal (t-4HN), a product of lipid peroxidation, reacted nonenzymatically with deoxyguanosine at pH 7.4 at 37 degrees C in vitro with each compound yielding two pairs of diastereomeric adducts. Adducts were isolated using reverse phase high-performance liquid chromatography and were characterized by their mass spectra and proton magnetic resonance spectra. Adducts 1 and 2 from t-4HH were assigned the structures 3-(2-deoxy-beta-D-erythro-pentofuranosyl)-5,6,7,8-tetrahydro-8R-hydroxy- 6S[1- (R and S)hydroxypropyl]pyramido[1,2-a]purine-10-(3H)one and Adducts 3 and 4 were assigned the structures 3-(2-deoxy-beta-D-erythro-pentofuranosyl)-5,6,7,8-tetrahydro-8S-hydroxy- 6R-[1- (R and S)hydroxypropyl]pyramido[1,2-a]purine-10-(3H)one. Similar 6-hydroxyhexyl adducts were isolated in the reaction of deoxyguanosine with t-4HN. The reactions appear to involve Michael additions of the N2 amino group of deoxyguanosine followed by cyclization at the 1-N site. This reaction mechanism is similar to that reported for deoxyguanosine adduct formation with the nonhydroxylated alpha, beta-unsaturated aldehydes crotonaldehyde and acrolein. Total adduct formations following 16-h incubations were 0.91% for t-4HH and 0.85% for t-4HN. These results demonstrate that t-4HH and t-4HN possess the ability to alkylate deoxyguanosine in vitro and suggest possible mechanisms for 4-hydroxyalkenal and pyrrolizidine alkaloid genotoxicity.

Topics: Aldehydes; Chemical Phenomena; Chemistry; Chromatography, High Pressure Liquid; Deoxyguanosine; Magnetic Resonance Spectroscopy; Mass Spectrometry; Mutagens