Oxidative profiles of LDL and HDL isolated from women with preeclampsia.

BACKGROUND: Oxidative stress causes biochemical changes in lipids and proteins; these changes can induce damage to the vascular endothelium and create maternal complications that are characteristic of preeclampsia. In this study, we evaluated the oxidative profile of lipoproteins isolated from women with preeclampsia. METHODS: Thirty women diagnosed with preeclampsia and thirty women without preeclampsia were included in the study. Lipid-damage biomarkers, including conjugated dienes, lipohydroperoxides and malondialdehyde, were measured. The reduction of nitroblue tetrazolium, the formation of dityrosines, and the carbonylation of proteins were assessed as indicators of protein damage. The protective activity of HDL-c was evaluated by the paraoxonase-I activity present on the HDL-c particles. Serum lipid profiles were also quantified in both groups. Data were analysed using Student's t test and the Pearson correlation coefficient. RESULTS: Our results demonstrated in PE women evident oxidative changes in the lipids and proteins in HDL-c and LDL-c particles and the activity of the antioxidant enzyme PON-I decreased 59.9%. HDL-c exhibited self-defence, as demonstrated by the negative correlation between paraoxonase-I activity and the formation of lipohydroperoxides in HDL-c (r = -0.3755, p < 0.005). CONCLUSIONS: LDL-c and HDL-c isolated from women with preeclampsia show oxidative damage to lipids and proteins. We propose an oxidative profile based on the oxidation levels indicated by each of the markers used. We also found that paraoxonase-I is inactivated in the presence of lipohydroperoxides. Antioxidant support might be helpful to reduce oxidative stress in patients with preeclampsia. Further investigations are necessary to define the association between antioxidant activities and preeclampsia.

Correlation of Plasma Protein Carbonyls and C-Reactive Protein with GOLD Stage Progression in COPD Patients.

Oxidative stress plays an important role in the pathogenesis of chronic obstructive pulmonary disease (COPD). To investigate the correlation between the progression of COPD and plasma biomarkers of chronic inflammation and oxidative injury, blood samples were obtained from healthy volunteers (HV, n = 14) and stabilized COPD patients. The patients were divided into three groups according to their GOLD stage (II, n = 34; III, n = 18; IV, n = 20). C-reactive protein (CRP), protein carbonyls (PC), malondialdehyde (MDA), susceptible lipoperoxidation of plasma substrates (SLPS), and myeloperoxidase activity (MPO) were measured. The plasma concentration of SLPS was measured as the amount of MDA generated by a metal ion-catalyzed reaction in vitro. PC, SLPS, and CPR were increased significantly (p < 0.001) in COPD patients when compared to HV. MDA concentrations and MPO activities were not significantly different from those of the HV group. In conclusion, increased oxidation of lipids and proteins resulting in a progressive increase in the amount of total plasma carbonyls and oxidative stress the presence of oxidative stress during COPD progression, concomitant with an increased oxidation of lipids and proteins resulting in a progressive and significant increase in the amount of total carbonyls formed from lipid-derived aldehydes and direct amino acid side chain oxidation in plasma, may serve as a biomarker and independent monitor of COPD progression and oxidative stress injury.

Glucose-stimulated acrolein production from unsaturated fatty acids.

Glucose auto-oxidation may be a significant source of reactive oxygen species (ROS), and also be important in the lipid peroxidation process, accompanied by the release of toxic reactive products. We wanted to demonstrate that acrolein can be formed directly and actively from free fatty acids in a hyperglycemic environment. A suspension of linoleic and arachidonic acids (2.5 mM) was exposed to different glucose concentrations (5, 10 and 15 mmol/L) in vitro. The samples were extracted with organic solvents, partitioned, followed at 255-267 nm, and analysed using capillary electrophoresis and mass spectroscopy. The total release of aldehydes significantly (P < 0.01) increased from 1.0 to 5.1, 8.3 and 13.1 micromol/L after 6 hours of incubation, proportional to glucose concentrations. It was possible to verify a correlate hydroperoxide formation as well. Among the lipid peroxidation products, acrolein (5% of total) and its condensing product, 4-hydroxy-hexenal, were identified. From the results presented here, it was possible to demonstrate the production of acrolein, probably as a fatty acid product, due to free radicals generated from the glucose auto-oxidation process. The results led us to propose that acrolein, which is one of the most toxic aldehydes, is produced during hyperglycemic states, and may lead to tissue injury, as one of the initial problems to be linked to high levels of glucose in vivo.

[Nitric oxide as main effector in the interleukin-1 system in ovulation]. / El óxido nítrico como principal efector del sistema de la interleucina-1 en la ovulación.

Ovulation is a complex process involving not only gonadotropins and steroid hormones, but also many local mediators common to inflammatory reactions, such as cytokines. Of particular interest is the ovarian interleukin-1 (IL-1) system, which may be an intermediary of gonadotropins in the ovulatory process. The preovulatory follicles have a complete and highly compartmentalized intraovarian IL-1 system including ligands, receptor, and receptor antagonist. IL-1 has been considered as the inductor of several ovulation-associated events such as prostaglandin and progesterone biosynthesis, plasminogen activator production, glycosaminoglycan generation, and enhancement of vascular permeability. The principal effector of the IL-1 system is nitric oxide. This paper analyzes the sites of synthesis and action of the IL-1 system in preovulatory follicle and its vascular dynamics as well as IL-1's mechanism of action in triggering follicular rupture.

Presence of uterine peroxidase activity in the rat early pregnancy.

Peroxidase has been associated with estrogen action in the uterus. This enzyme plays an important role in the control of hydrogen peroxide levels and in catechol estrogen production. Since the uterus, during early pregnancy, is subjected to estrogen and progesterone regulation, we analyzed the changes of peroxidase activity in relation to receptivity and uterine early response to the embryo. Soluble and microsomal peroxidase activity were determined in the rat uterus during the estrus phase and early pregnancy (days 3 through 6). Soluble peroxidase activity increased significantly (p < 0.01) from day 3 (1.50 +/- 0.24) to day 4 (3.5 +/- 0.3) and 5 (5 +/- 0.5 U/mg protein, mean +/- S.D., n = 6) of pregnancy. During day 6, a significant decrease was noted in both the implantation site and the nonimplantation uterine tissue. Microsomal calcium-extractable peroxidase showed a similar pattern, with lower specific activity than, the soluble peroxidase. During estrus, the uterine tissue showed the highest activity of calcium-extracted peroxidase (8.7 +/- 1.35 U/mg protein), statistically greater when compared with days 3, 4, 5 and 6 of pregnancy. In conclusion, high peroxidase activity was associated with uterine receptivity. The decrease of activity on day 6 might be due to a progesterone-estrogen interaction, and consequently, hydrogen peroxide can be utilized for hydroxile production by means of the Fenton reaction. Lipoperoxidation may be necessary for changes in membrane fluidity for embryo attachment to endometrial epithelium.

Pro-oxidating properties of melatonin in the in vitro interaction with the singlet oxygen.

In an aqueous system, the oxidation of the erythrocyte membrane by the singlet oxygen formed during the photoactivation of the rose bengal coloring was examined. The effects of the singlet oxygen on lipids and proteins were studied through the simultaneous quantification of peroxidation products, lipoperoxides and carbonyl groups, the oxidation of protein SH groups and the activity of the glyceraldehyde 3-phosphate dehydrogenase (G3PD) associated with the erythrocyte membrane. The antioxidant activity of melatonin was tested and compared to that of two antioxidants in extreme cases of hydrosolubility, ascorbate and beta-carotene, with the purpose of comparing the protective ability of melatonin against singlet oxygen. The results show the expected effect even at low (0.125-0.75 mM; 0.015-0.90 mM, respectively) for ascorbate and beta-carotene, antioxidants known to possess important antioxidant qualities against singlet oxygen. It is shown that melatonin, under the conditions described, and at the concentrations at which the other two compounds are efficacious, not only confers little antioxidant protection, but that a pro-oxidant tendency was proven both on lipids and proteins, as well as on G3PD enzymatic activity. The results show that the antioxidant protective effect that melatonin can exert on biological systems is probably not by a direct interaction with oxidant species, but probably, as has been previously proposed, through the regulation of antioxidant defense systems. The formation of secondary oxidation products, such as melatonin-derived endoperoxides, may explain the evidence found on pro-oxidant qualities of this molecule.

Identification of acrolein from the ozone oxidation of unsaturated fatty acids.

By-products of lipoperoxidation reactions may be associated with the genesis or the progression of several diseases as arteriosclerosis, diabetes and cancer, among many others. Acrolein, at first a widely distributed environmental pollutant, is currently known as a compound capable of being generated as a result of metabolic reactions within biological systems, highly toxic and the most electrophilic of the alpha, beta-unsaturated aldehydes formed during lipoperoxidation. In the present study: 1. The separation of acrolein and malondialdehyde was achieved at alkaline pH with the use of high voltage capillary electrophoresis in uncoated fused-silica capillaries. 2. It was demonstrated how the oxidation of fatty acids (arachidonic/linoleic) with ozone generates, in dose-dependent form, acrolein as one of the by-products of the lipoperoxidation process. The oxidation of open human erythrocyte membranes with ozone also generated acrolein. 3. After aldolic condensation, aldol-acrolein derivative has a positive reaction with 2-thiobarbituric acid (TBA) and shows a maximum absorption at 498 nm. This novel characteristic is used in its identification after the separation of the by-products. 4. It is possible to suggest that in the classic reaction of the denominated thiobarbituric acid reactive substances (TBARS), when used as an indicator of the degree of peroxidation in biological systems, a portion of acrolein could be present but dwarfed by the TBA-MDA adduct.

Oxidation of gonadotrophin (PMSG) by oxygen free radicals alters its structure and hormonal activity.

The effect of oxygen free radicals produced by the Fenton reaction was used to induce oxidation and other structural changes in pregnant mare serum gonadotrophin (PMSG). Modifications in the spectrophotometric scan, an increase in exposed carbonyl groups, and the ability to reduce nitroblue tetrazolium, was achieved by the oxidized hormone when compared to the control PMSG. PMSG loses its biological activity when coming in contact with the free-radical generating system. This lack of activity is manifested as a loss of ovulation and a decrease in the weight of the ovaries and uterus. It was demonstrated that oxygen free radicals can induce structural and biological changes in the gonadotrophin.

Effect of nitric oxide releasers on some metabolic processes of rabbit spermatozoa.

The authors investigated the possible effect of nitric oxide (NO) releasers (the free radical form of nitrogen monoxide, which control some functions of many cells) on rabbit spermatozoa. A significant (P < .01) increment was found in the percentage of the acrosome reaction in rabbit spermatozoa incubated for 30-60 min in presence of the NO releasers sodium nitroprusside (SNP) and N-acetyl-S-nitroso cysteine (NACysSNO), but not with S-nitroso cysteine (CysSNO). This effect was reverted or lowered when the NO scavenger HbO2 was included in the medium. The effects of SNP and NACysSNO on acrosome reaction do not appear to be related to glucose utilization, viability, or lipid peroxidation.

Uterine glutathione reductase activity: modulation by estrogens and progesterone.

The aim of this study was to determine whether glutathione reductase activity in uterine tissue is regulated by sex hormones. In spayed rats uterine glutathione reductase was significantly increased by exogenous estrogen (P< 0.01), progesterone (P< 0.01) or estrogen plus progesterone (P<0.01). When enzyme activity is expressed per mg protein, daily administration of estrogen or progesterone induces a progressive increase of this enzyme between 24 to 48 h or 24 to 72 h of treatment, respectively. Whereas the combination of both steroids causes an earlier and higher increase in glutathione reductase activity at 24 h of treatment. Estradiol singly or in combination with progesterone induced the highest protein concentration in the uterus. Whereas uterine DNA concentration is only significantly affected by estradiol. Our results suggest that uterine glutathione reductase is regulated by estradiol and progesterone and may be involved in maintaining levels of reduced glutathione in the uterus. This compound may be required for control of the redox state of thiol groups and in detoxification reactions involving H2O2 and electrophylic substances. The antioxidant action of estrogens is partially due to the stimulation of glutathione reductase.

Changes in human serum antioxidant capacity and peroxidation after four months of exposure to air pollutants.

Twenty-one adult volunteers (aged 27-32 years), who had been living in Mexico City for four continuous months (physicians working as fellows) were studied the first and sixteenth week of their stay in order to learn the effects of the pollutants contained in Mexico City's atmosphere on some serum biochemical parameters. The activity of serum superoxide dismutase (SOD) decreased after 16 weeks in comparison with the values obtained the first week (109.6 to 56.9 mU/mg protein; 50% less). In contrast, the inhibitory capacity of serum vs. induced in vitro lipoperoxidation increased in relation to the length of stay (22%). The serum levels of thiobarbituric-reactive material also decreased in almost 30% (from 6.10 to 4.12 nmol). The other lipoperoxides measured were unchanged (chromolipids and diene conjugation). We propose that this may be as a result of the adaptative capacity of the human organism, within a pollutant atmosphere in which the ozone levels might participate in a decrease of SOD activity during chronic exposure, to air pollution.

Effect of colchicine on nuclear presence of two lysosomal enzymes in rat implantation sites.

The purpose of the study was to demonstrate that the effect of colchicine on the implantation and embryo development in rat is caused in part by its action on lysosome translocation to the perinuclear region. The subcellular enzymatic distribution of two lysosomal enzymes, acid phosphatase (E.C.3.1.3.2.) and beta-glucuronidase (E.C.3.2.1.3.1.), were measured. The right uterine horn was treated during preimplantation with colchicine (2 micrograms/kg of body weight on day 4) and the left with lumicolchicine (control). In the control horn, the nuclear activity of lysosomal enzymes was significantly higher in the implantation site tissue than the treated horn (colchicine) (p < 0.01). There were no modifications on the undecidualized endometrium under colchicine treatment. Simultaneously to this result, implantation and embryonic development were abolished. From the results presented herein, it is proposed that the inhibitory effect of colchicine is due to an inadequate biochemical differentiation at the implantation site, related to both arrest of cell division (mitosis is arrested in methaphase) and inhibition of the lysosomal movement toward the nucleus.

[New endometrial modulators in early pregnancy]. / Los nuevos moduladores endometriales en el embarazo temprano.

Cytokines synthesized by the uterus or placenta include those thought to be produced exclusively by, or though to act on, cells of the lymphohematopoietic system. Although many of these cytokines are protein mediators of the immune system effector phase, in the female reproductive tract their principal target cells and sites of synthesis are non-lymphohematopoietic cells. During pregnancy, uterine epithelial cells, decidual cells and trophoblast appear to be major sources of the classic lymphohematopoietic cytokines. This suggests two not necessarily exclusive alternatives: that these cells are extensions of, or are involved in, regulating the immune system, or that these factors regulate growth and differentiation of uterine and embryonic tissues. This paper analyzes the sites of synthesis, targets and possible functions of the cytokines during early pregnancy.

Participation of oxygen-free radicals in the oxido-reduction of proteins.

Recent studies have focused attention on the possible role of active oxygen species on protein damage and degradation. The reactions of free radicals on biomolecules are important in physiology and pathology. A number of systems that generate free radicals catalyze the oxidative modification of proteins in two species: protein peroxides, which can consume important antioxidants; and protein-bound reducing moieties, which can reduce transition metals, and may enhance their activity in radical reactions. Protein oxidation also contributes to the pool of damaged enzymes and accumulation of abnormal and damaged proteins, which increases during aging and in various pathological states, such as atherosclerosis, cancer, etc.

Possible effect of air pollutants (Mexico City) on superoxide dismutase activity and serum lipoperoxides in the human adult.

The action of air pollutants, through their constituents, (O3, NO2, tobacco smoke) are capable of causing damage due to their lipoperoxidative properties or, indirectly, by inducing production of free radicals. As a consequence of photochemical processes, the ozone levels in the atmosphere of Mexico City are generally higher (mean of 0.325 ppm; period between 1987-1992) and may be harmful to health. Sixty two volunteers (medical doctors), aged 27-32 years, were divided into three groups. Group A was composed of those persons (17) who had never lived in Mexico City; a second group (B) (21) had recently arrived in Mexico City (1-8 days); and a third group (C) (24) who had permanently resided in Mexico City. Serum was obtained from fresh whole blood. Superoxide dismutase (SOD) activity and thiobarbituric acid-reactive materials were higher in group B while chromolipids and the serum inhibitory capacity (for lipoperoxidation) was higher in group C. The acute exposure to pollutants in group B apparently may have induced SOD as an antioxidant defense and was responsible for the increased level of TBA reactive material. In group C, the significant finding is better antioxidative defenses and slightly higher chromolipids.

[Participation of the cytoskeleton in the physiology of the endometrium]. / Participación del citoesqueleto en la fisiología del endometrio.

The cytoskeleton in the endometrium, takes part not only in all the mechanic functions of the cell, but because of movement and location of healthy organelles and proteins, it also takes part in the metabolism. The endometrial epithelium, because of its morphology and its supposed cellular homogeneity, has been studied more than the stroma. It is known that intermedium filaments show a characteristic pattern of typical distribution and expression of the cellular type. During pregnancy and pseudopregnancy, in the apical region of the epithelial cells, both, luminal and glandular, there is an abundance of keratin in the basolateral region; while the vimentin is abundant only in the luminal epithelial cells and it increases in the implantation day. In humans and rats, the desmin only expresses during the decidual response. It is considered that intermedium filaments have a role in the polarity changes of the membrane. The microfilaments (MF) are related with the regulation of the cellular morphology and movement. In the luminal epithelium the MF play a role in the transformations of the uterine surface like the microvilli. The microtubules in the endometrium and other organs play an important role in the organelles position like lysosomes, mitochondria and Golgi complex. Also it is proved that take part in the DNA synthesis, because colchicine drug inhibits it.

[Lysosomal system in hormonal mechanisms. Review]. / El sistema lisosomal en los mecanismos hormonales. Revisión.

The role of lysosomes in the intracellular mechanism of action of several steroid an proteic hormones has been demonstrated. In presence of the specific hormone the target cell induce membranal changes and the lysosomes are moved toward the nucleus; after this the lysosomal enzymes are released in the perinuclear space. For the moment it is not possible to know the biochemical role of this enzymatic activities upon the nucleic acids function and des-repretion process of specific genes, but the inhibition of lysosomes movement utilizing hormone antagonist or dexamethasone inhibits some reproductive process like the implantation of the mammalian egg. We present herein a review related with the mode action of some hormones through the lysosomes in reproductive processes.

Inhibitory capacity of human serum on induced microsomal lipoperoxidation.

The capacity of human serum for inhibiting in vitro the membrane lipoperoxidation induced by a controlled system (ADP/NADPH + H+/Fe3+) was demonstrated. A concentration of 8 nmol of malondialdehyde was produced in 20 min in rat liver microsomes (1.5 mg of protein) after exposure to an induced lipoperoxidation mixture. Addition of 100 microliters (13.89 mg of protein) of human serum decreased malondialdehyde production nearly 50%. An increase of 25.97% of the inhibitory capacity of serum was obtained by the in vitro addition of 10 microliters/ml of vitamin E. Ten volunteers were supplemented with 400 mg of vitamin E and 1 g of vitamin C/daily for 2 weeks. Their serum inhibitory capacity increased in 12% (p < 0.05). The serum inhibitory capacity for microsomal lipoperoxidation is described herein, and we propose its utilization as an index to determine the individual nonspecific antioxidative defenses against free radical injury and lipoperoxidation in relation to exposure to air pollutants, tobacco smoke, and several acute and chronic diseases, including the hypoxia-reperfusion phenomena.

Effect of dexamethasone as an inhibitor of implantation and embryo development in rat; lysosomal role.

In order to learn the mechanism of action of dexamethasone administration as an efficient inhibitor of estrogen activity in different tissues, the subcellular enzymatic distribution of two lysosomal enzymes: acid phosphatase (E.C.3.1.3.2.) and beta glucuronidase (E.C.3.2.1.3.1.) were measured. The rats were treated during preimplantation with dexamethasone (0.8 mg on days 3 and 4) or saline (controls). In the control group, the nuclear activity of lysosomal enzymes was significantly less in the implantation site tissue than in the treated group (p < 0.05). There were no modifications on the undecidualized endometrium under the steroid treatment. The lysosomal subfraction showed an opposite response. The steroid treatment produced an increase of activity in the decidualized tissue (1.9 +/- 0.4 to 4.9 +/- 0.4) while the nuclear enzymatic activity decreased under treatment; and simultaneously, the embryonic development was 100% abolished. From the results presented herein, it is proposed that the inhibitory effect of dexamethasone upon implantation is due to an inadequate biochemical differentiation at the implantation site, related to the inhibition of lysosomal movement toward the nucleus, and consequently to lysosomal enzymatic release and metabolic role.