Diaphragmatic lipid peroxidation in chronically loaded rats

Since the functional outcome of effector T lymphocytes depends on a balance between activatory and inhibitory receptors, we studied the ability of CTLA-4 (CD152) to inhibit the cytolytic function of CTL. In 22 TCR alpha/beta+ CD3+ 8+ CTL clones, activation induced by anti-CD3, anti-CD28, or anti-CD2 mAb was inhibited by anti-CD152 mAb in a redirected killing assay. In eight clones inhibition was >40%, in 10 it ranged between 20-40%, and in four it was <20%. This suggests the existence of a clonal heterogeneity as well as for the ability of CTLA-4 to inhibit CD3/TCR-, CD28-, or CD2-mediated CTL activation. To support further this contention, we used an experimental model based upon Ag-specific CTL. Eight Ag-specific T cell clones that lyse autologous EBV-infected B lymphocytes, but are unable to lyse allogeneic EBV-infected B cell lines, were used in a cytolytic assay in which anti-CD152 mAb or soluble recombinant receptor (i.e., CTLA-4 Ig) were included. In this system, at variance from the redirected killing assay, cross-linking of surface molecules by mAb does not occur. Thus, addition of anti-CD152 mAb or of CTLA-4 Ig and anti-CD80/CD86 mAb to the assay should result in a blockade of receptor/ligand interactions. As a consequence, inhibition of a negative signal, such as that delivered via CD152, should enhance lysis. A >40% increment of target cell lysis was achieved in three of eight clones studied. Since it is not equally shared by all CTL clones, this feature also appears to be clonally distributed.     

Endocrine interactions in regulation of body weight by female rats.

Studied food intakes and body weights in 65 overweight female Sprague-Dawley rats that were sham-operated, adrenalectomized, or adrenalectomized and ovariectomized. Before surgery the groups were equivalent. Postoperatively, the adrenalectomized Ss showed diminished food consumption and body weight relative to the ovariectomized and adrenalectomized Ss. Data are discussed in terms of a delicate estrogen-progesterone balance in the control of body weight. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Ovarian-adrenal interactions in regulation of body weight by female rats.

Conducted 7 experiments with a total of 40 female Sherman albino rats. Ovariectomy induced an increase, and estrogen treatment after ovariectomy a decrease, in the level at which Ss regulated their body weights. Food-intake changes were transient, intake returning toward normal values as terminal body weight was approached. Results suggest these intake changes were probably secondary to effects on the weight-regulating system itself. Adrenalectomy blocked or reversed the weight gain which followed ovariectomy. This effect was not a matter of incapacity but seemed to reflect a resetting of the weight-regulating system so that a low weight level was voluntarily maintained. (20 ref.) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Study on body mass index, lipid profile and lipid peroxidation status in coronary artery disease

Among the many known risk factors of coronary artery disease (CAD) obesity and hypercholesterolaemia are important ones. Whatever may be the risk factor, the basic pathology of CAD is deposition of altered lipids on the endothelium. One of such altered lipid is oxidatively modified low density lipoprotein (LDL). Lipid peroxidation has been assessed by several methods. Quantitation of malondialdehyde (MDA) by thiobarbituric acid (TBA) method is one of the commonly utilised method in several laboratories. In this study 40 cases of CAD were selected for evaluation. The body mass index (BMI), lipid profile and the level of lipid peroxidation (MDA) were measured. Seventeen cases (42.5%) had normal BMI (20-25), 20 cases (50%) were in the overweight range of BMI (26-30) and only 3 cases (7.5%) were in the obese group with a BMI more than 30. BMI correlated better with the level of total cholesterol (Tc), low density lipoprotein cholesterol (HDLc) and MDA. BMI did not show any correlation with triglyceride (Tg) or high density lipoprotein cholesterol (HDLc). MDA level correlated better with Tc, Tg levels and BMI, poorly correlated with LDLc and in inverse relationship was observed with HDLc.     

Progesterone protects against lipid peroxidation following traumatic brain injury in rats

Large scale use of lysozyme for periplasmic release has been impeded by the cost of the pure enzyme and its subsequent presence as a contaminant in later downstream processing steps. In this paper, we discuss the use of lysozyme for pilot scale recovery of a periplasmic enzyme from E. coli. The effects of concentration of sucrose, lysozyme and cells on periplasmic enzyme release were examined. Lysozyme concentration can be reduced 5-fold from previous reports and a reduction in sucrose concentration from 20 to 15% (w/v) allows an improvement in centrifugal harvesting by reducing viscosity. High levels of release were still achieved using this technique and further improvements in yield were obtained by optimising other components of the releasing mixture. Results show that some release is still achieved in circumstances where no lysozyme use is possible. Results also indicate that a substantial proportion (up to 70%) of lysozyme remains bound to the cellular debris after its action and is removed with this material.     

Inhibition of lipid peroxidation mediated by indolizines

Esters, ethers, carbonates and carbamates of 1-indolizinols and azaindolizinols exhibit a profound inhibition of lipid peroxidation in vitro. The antioxidants were prepared by cyclization of pyridines and diazines with diphenylcyclopropenone followed by introduction of the O-substituent.     

Prevention of carbon tetrachloride-induced lipid peroxidation in liver microsomes from dehydroepiandrosterone-pretreated rats

Dehydroepiandrosterone (DHEA), a lipid soluble steroid, administered to rats (100 mg/kg b.wt) by a single intraperitoneal injection, increases to twice its normal level in the liver microsomes. Microsomes so enriched become resistant to lipid peroxidation induced by incubation with carbon tetrachloride in the presence of a NADPH-regenerating system: also the lipid peroxidation-dependent inactivation of glucose-6-phosphatase and gamma-glutamyl transpetidase due to the haloalkane are prevented. Noteworthy, the liver microsomal drug-metabolizing enzymes and in particular the catalytic activity of cytochrome P450IIE1, responsible for the CCl4-activation, are not impaired by the supplementation with the steroid. Consistently, in DHEA-pretreated microsomes the protein covalent binding of the trichloromethyl radical (CCl3 degrees), is similar to that of not supplemented microsomes treated with CCl4. It thus seems likely that DHEA protects liver microsomes from oxidative damage induced by carbon tetrachloride through its own antioxidant properties rather than inhibiting the metabolism of the toxin.     

Promotion of colon carcinogenesis through increasing lipid peroxidation induced in rats by a high cholesterol diet

Recent studies suggest a role of the neural cell adhesion molecules L1 and NCAM in mechanisms of memory storage. In the present study we analyzed the effect of continuous intraventricular infusion of polyclonal antibodies directed against L1 (antiL1) or NCAM (antiNCAM) on the performance of male Wistar rats during the acquisition and retention of a spatial learning task (Morris water-maze). In this task animals have to learn the spatial position of a hidden escape platform in a water tank to escape onto it. During acquisition of the task animals with continuous infusion of antiNCAM - but not those infused with antiL1 - showed day-dependent attenuated learning in comparison to controls (P = 0.001). Control animals were either injected with vehicle (PBS) or with polyclonal antibodies raised against liver cell membrane. When the escape platform was removed during the retention test (transfer test), the performance of animals continuously infused with antiL1 as well as those continuously infused with antiNCAM showed an impaired search pattern when compared with the performance of control animals (P = 0.001 and 0.04, respectively). Whereas control animals spent up to 46% of their time searching for the platform in the correct quadrant, the time antiL1- and antiNCAM-infused animals spent in this quadrant was closer to chance level (30.5% and 36.5%), respectively). The present data provide additional support for an involvement of the two adhesion molecules L1 and NCAM in synaptic plasticity underlying memory storage.     

Effect of hyperbaric oxygen on lipid peroxidation in free skin grafts in rats

OBJECTIVE: To document that free skin grafts treated with hyperbaric oxygen (HBO) are at greater risk for reperfusion injury, resulting in lipid peroxidation, than are free skin grafts without HBO treatment. ANIMALS: 40 Sprague-Dawley rats. PROCEDURE: Free skin grafting was performed bilaterally on each rat. The HBO-treated rats received HBO twice daily for 90 minutes at 2 ATA. Biopsy specimens were taken from each rat at the time of grafting and on days 2, 4, 7, 10, 14, 21, and 28, then were processed for tissue concentration of total glutathione (GSHt), glutathione peroxidase activity (GPx), and presence of thiobarbituric acid-reactive substances (TBARS). RESULTS: Both groups had a similar pattern of change in TBARS and GPx values--initial increase, returning to preoperative values at days 21 (control) and 28 (HBO). The GPx activity peaked later than did TBARS concentration (day 7 vs day 4). The pattern was significantly more pronounced in HBO-treated than in control rats. Both groups had a similar pattern of change in GSHt values-significant decrease from preoperative concentration at day 2, return to preoperative concentration by days 4 (HBO) and 7 (control), increase above preoperative concentration by day 21, and return to preoperative concentration by day 28. Obvious visual or histologic differences in the grafts were not detected between groups. CONCLUSIONS: Cellular effects of oxidative stress were apparent in both groups of rats; however, the degree of these effects was exacerbated by HBO. In the face of enhanced cellular lipid peroxidation, use of HBO for the treatment of free skin grafts must be questioned.     

Measurement of lipid peroxidation

Lipid peroxidation results in the formation of conjugated dienes, lipid hydroperoxides and degradation products such as alkanes, aldehydes and isoprostanes. The approach to the quantitative assessment of lipid peroxidation depends on whether the samples involve complex biological material obtained in vivo, or whether the samples involve relatively simple mixtures obtained in vitro. Samples obtained in vivo contain a large number of products which themselves may undergo metabolism. The measurement of conjugated diene formation is generally applied as a dynamic quantitation e.g. during the oxidation of LDL, and is not generally applied to samples obtained in vivo. Lipid hydroperoxides readily decompose, but can be measured directly and indirectly by a variety of techniques. The measurement of MDA by the TBAR assay is non-specific, and is generally poor when applied to biological samples. More recent assays based on the measurement of MDA or HNE-lysine adducts are likely to be more applicable to biological samples, since adducts of these reactive aldehydes are relatively stable. The discovery of the isoprostanes as lipid peroxidation products which can be measured by gas chromatography mass spectrometry or immunoassay has opened a new avenue by which to quantify lipid peroxidation in vivo, and will be discussed in detail.     

Role of active oxygen species and lipid peroxidation in mepirizole-induced duodenal ulcers in rats

The role of active oxygen species and lipid peroxidation in the pathogenesis of duodenal ulcers induced by mepirizole was investigated in rats. Oral administration of mepirizole (200 mg/kg) resulted in ulcer lesions in the proximal duodenum. Thiobarbituric acid-reactive substances (TBA-reactive substances), an indicator of lipid peroxidation, also significantly increased in the duodenal mucosa. Myeloperoxidase (MPO) activity in the duodenal mucosa, a sign of polymorphonuclear leukocyte (PMN) accumulation, significantly increased. Combination treatment with polyethylene glycol-modified Serratia Mn-SOD and catalase significantly decreased the size of the ulcers and TBA-reactive substances in the duodenal mucosa. Allopurinol, a xanthine oxidase inhibitor, also reduced the size of duodenal ulcers. Both the size of the ulcers and the increase in TBA-reactive substances in the duodenal mucosa were significantly lower in PMN-depleted rats. Mepirizole increased the surface expression of adhesion molecule CD18 on PMNs in vitro. These results suggest that lipid peroxidation, mediated by active oxygen species generated from xanthine oxidase and PMNs, plays an important role in the pathogenesis of duodenal ulcers induced by mepirizole.     

Crosslinking of apolipoprotein E by products of lipid peroxidation

Apolipoprotein E (APOE) genotype and advancing aging are interacting ri sk factors in the expression of late onset and sporadic Alzheimer's Disease (AD). We tested the hypothesis that 2 products of lipid peroxidation, malondialdehyde (MDA) and 4 hydroxy-2-nonenal (HNE), covalently modify APOE and alter its metabolism. In vitro, both HNE and MDA crosslinked purified APOE3 and APOE4. HNE was a more potent crosslinker than MDA, and purified APO3 was more susceptible to crosslinking by HNE than was purified APOE4. In P19 neuroglial cultures, oxidative stress with lipid peroxidation led to increased intracellular accumulation of anti-HNE and anti-APOE immunoreactive proteins of approximately 50 kDa. Intercellular accumulation of the 50 kDa APOE-immunoreactive protein (APOE-50) was not prevented by cyclohexamide, suggesting formation by post-translational mechanisms. In CSF, a 50 kDa APOE-immunoreactive protein co-migrated with proteins most immunoreactive for HNE and MDA adducts, containing NaB3H4-reducible bonds. These proteins were in CSF from adult subjects (with or without dementia), and in AD patients homozygous for APOE3 or APOE4 alleles. These data suggest that HNE covalently crosslinks APOE in P19 neuroglial cultures to form a 50 kDa protein, and that similar modifications of APOE appear to occur in vivo.     

Dietary flavonoids reduce lipid peroxidation in rats fed polyunsaturated or monounsaturated fat diets

We investigated the influence of dietary flavonoids on alpha-tocopherol status and LDL peroxidation in rats fed diets enriched in either polyunsaturated fatty acids (PUFA) or monounsaturated fatty acids (MUFA). Diets equalized for alpha-tocopherol concentrations were or were not supplemented with 8 g/kg diet of flavonoids (quercetin + catechin, 2:1). After 4 wk of feeding, plasma lipid concentrations were lower in rats fed PUFA than in those fed MUFA with a significant correlation between plasma alpha-tocopherol and cholesterol concentrations, r = 0.94, P < 0. 0001). Dietary lipids influenced the fatty acid composition of VLDL + LDL more than that of HDL or microsomes. The resistance of VLDL + LDL to copper-induced oxidation was higher in rats fed MUFA than in those fed PUFA as assessed by the lower production of conjugated dienes and thiobarbituric acid reactive substances (TBARS) and by the >100% longer lag time for dienes production. (P < 0.0001). Dietary flavonoids significantly reduced by 22% the amounts of dienes produced during 12 h of oxidation in rats fed diets rich in PUFA and lengthened lag time 43% in those fed MUFA. Microsomes of rats fed MUFA produced approximately 50% less TBARS than those of rats fed PUFA (P < 0.0001) and they contained more alpha-tocopherol in rats fed MUFA than in those fed PUFA with higher values (P < 0. 0001) in both groups supplemented with flavonoids (P < 0.0001). Our findings suggest that the intake of dietary flavonoids is beneficial not only when diets are rich in PUFA but also when they are rich in MUFA. It seems likely that these substances contribute to the antioxidant defense and reduce the consumption of alpha-tocopherol in both lipoproteins and membranes.     

Effect of alpha-tocopherol on lipid peroxidation and total antioxidant status in spontaneously hypertensive rats

The aim of this study was to determine the effects of alpha-tocopherol on lipid peroxidation and total antioxidant status of spontaneously hypertensive rats (SHR), comparing them with normal Wistar-Kyoto (WKY) rats. SHR were divided into three groups and treated with different doses of alpha-tocopherol (alpha1, 17 mg/kg diet; alpha2, 34 mg/kg diet; and alpha3, 170 mg/kg diet). Normal WKY and untreated SHR were used as normal (N) and hypertensive control (HC). Blood pressures were recorded every 10 days for 3 months. At the end of the trial, animals were killed and measurement of plasma total antioxidant status, plasma superoxide dismutase (SOD) activity, and lipid peroxide levels in plasma and blood vessels was carried out following well-established methods. From our study it was found that lipid peroxides in thoracic aorta (N, 0.47 +/- 0.17; H, 0.96 +/- 0.37; P < .0001) and plasma (N, 0.06 +/- 0.01; H, 0.13 +/- 0.01) were significantly higher in hypertensives than in normal rats. SOD activity was significantly lower in hypertensive than normal rats (N, 172.93 +/- 46.91; H, 110.08 +/- 14.38; P < .005). Total antioxidant status was significantly higher in normal than hypertensive rats (N, 0.88 +/- 0.05; H, 0.83 +/- 0.02; P < .05). After the antioxidant trial, it was found that in the treated groups rise of blood pressure was prevented significantly (P < .001) and lipid peroxides in blood vessels were significantly reduced more than in the controls (P < .001). For plasma lipid peroxide it was only significant for groups alpha2 (P < .001) and alpha3 (P < .05). Although all three treated groups showed improved total antioxidant status, only groups alpha2 (0.87 +/- 0.04, P < .005) and alpha3 (1.20 +/- 0.18, P < .001) were statistically significant. All the three groups showed significant increases in their SOD activity (P < .001). Correlation studies showed that total antioxidant status and SOD were significantly negatively correlated with blood pressure in normal rats (P = .007; P = .008). Lipid peroxides in both blood vessel and plasma showed a positive correlation. In the treated groups, lipid peroxides in blood vessels maintained a significant positive correlation with blood pressure in all groups (alpha1, P = .021; alpha2, P = .019; alpha3, P = .002), whereas for plasma lipid peroxides the correlation was in groups alpha1 (P = .005) and alpha2 (P = .009). For SOD activity, significant negative correlations were found with blood pressure in the alpha2 (P = .017) and alpha3 (P = .025) groups. Total antioxidant status maintained a significant negative correlation with blood pressure in all three groups (alpha1, P = .012; alpha2, P = .044; alpha3, P = .014). In conclusion it was found that supplement of alpha-tocopherol may prevent development of increased blood pressure, reduce lipid peroxides in plasma and blood vessels, and enhance the total antioxidant status, including SOD activity.     

Enhanced lipid peroxidation in hepatic cirrhosis

> Objective: This paper describes the deliberations of an interdisciplinary group of clinical and basic scientists who met at the National Institute of Child Health and Human Development to discuss the potential role of fetal behavior in assessing fetal well being and predicting neonatal outcome. The conference focused on three aspects of fetal behavior: 1) habituation; 2) state transitions; and 3) movement. Methods: The participants consisted of 25 leaders in the fields of obstetrics, perinatal medicine, neonatology, developmental psychobiology, developmental neuroscience, developmental psychology, ethology, and mathematics. The meeting was divided into three parts. In each of these a plenary speaker (a recognized expert in his field) began the session with an overview of the scientific theme. Two respondents, with research expertise in fetal research (animal models or human fetuses) followed with remarks on the plenary talk and comments based on their own studies. At the conclusion of these comments, the participants met in small groups to discuss the plenary proceedings and their implications for assessing human fetal well being and predicting outcome. At the conclusion of the small group deliberations all of the participants reconvened in a plenary session. During this part of the meeting a rapporteur from each small group summarized their discussions. Results and Conclusions: 1) Fetal habituation: there was a general consensus that research on this aspect of fetal behavior may have a high payoff for assessing human fetal well being and predicting neonatal outcome. 2) Behavioral state transitions: participants agreed that transitions afford investigators with an indication of when (timing) and how (models) behavior changes within and between developmental periods. Knowledge of transitions during development allows for tracking of behaviors that may be necessary for the fetus to adapt to its in utero environment or prepare for its postnatal life. 3) Chaos theory and fetal movement: participants concluded that non-linear dynamics systems analysis models could be useful to analyze "noise" within a measurement system; better define time scales; and increase resolution and thereby better identify "signals."     

Inhibition of in vitro lipid peroxidation by stable steroidic nitroxyl radicals

4',4'-dimethylspiro (5 alpha-cholestane-3,2'-oxazolidin)-3'-yloxy (IK-1) and 7 alpha,12 alpha-dihydroxy-4',-4'-dimethylspiro (5 beta-cholan-24-oic-3,2'-oxazolidin)-3'-yloxy acid (IK-2), two stable steroidic nitroxyl radicals, were newly synthesized and tested as possible inhibitors of lipid peroxidation, induced by Fenton's reagent in both rat liver microsomes and egg phosphatidylcholine liposomes. The inhibitory activity, evaluated through the formation of thiobarbituric acid reactive substances (TBARS) and the conjugated diene, was compared with that of alpha-tocopherol and 2,2,6,6-tetramethylpiperidine-1-yloxy (TEMPO). In each model system IK-1 and IK-2 exhibited an IC50 of 8 microM and reduced the formation of TBARS and conjugated diene, showing IK-1 a potency comparable to alpha-tocopherol and higher than TEMPO. Moreover IK-1 and, to a lesser extent IK-2, reduced the lipid peroxidation induced in the microsomes by the water-soluble azo-initiator 2,2'-Azobis (2-methylpropionamidine) dihydrochloride (AMPH), indicating the IK-1 and IK-2 ability as chain-breaking antioxidants. The hydroxylamine 4',4'-dimethylspiro (5 alpha-cholestane-3,2'-oxazolidin)-3'-hydroxide (IK-3), obtained by chemical reduction of IK-1, was completely inactive as an inhibitor of lipid peroxidation in heat pre-treated microsomes and in liposomes. However in microsomes it was active since it was oxidized to the corresponding nitroxyl radical IK-1.     

Liver lipid accumulation and performance of hens as affected by cage density and initial body weight

Hens housed in individual cages (25 46 cm.) laid significantly more eggs, consumed significantly more feed and had significantly larger livers with a higher lipid content than hens housed three to a cage in two experiments. Body weight gain was significantly higher for individually housed birds in experiment 1, but not in experiment 2. No difference was observed in mean egg weight or kg. feed per dozen eggs. In experiment 2 hens housed two to a cage laid slightly fewer eggs and accumulated slightly less liver fat but the differences were not significantly different fromthose housed individually. Hens socially dominant in three bird pens had higher liver fat accumulation than hens lower on the peck order but liver fat accumulation for the dominant hens still averaged less than hens housed either two or one per cage. Comparison of two strains in experiment 1 revealed a significant difference in rate of egg production and feed efficiency but no difference in liver fat accumulation. Pullets placed in four body weight classes prior to the start of the experiment did not differ significantly in liver fat accumulation per unit of body weight or percentof dry matter of the liver at the end of the experiment. Rate of egg production and feed efficiency were also not significantly different among the body weight classes.