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.

Clear-air turbulence: simultaneous observations by radar and aircraft.

Ultrasensitive radars and uninstrumented jet aircraft in concert have probed regions of the clear atmosphere in search of clear-air turbulence. All sources of clear-air radar echoes above 6 kilometers that were probed simultaneously by the aircraft were found to be turbulent.

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.

Uterine estrogen sulfatase activity at the time of blastocyst implantation in the rat.

The conversion of estrone sulfate (E1S) to estrone (E1) was measured during the in vitro incubation of the labeled sulfoconjugate with implantation sites (IS) and nonimplanted regions (NIS) of uterine horns from 6-day pregnant rats. Extensive metabolism of E1S occurred in both tissues, being noticeably less (29.31%) in IS than in NIS. Estrogen sulfatase activity present in the uterus of ovariectomized virgin rats was found to be higher than in both uterine regions of the pregnant rats. We suggest that E1S present in uterine fluids may be accessible to be metabolized into unconjugated estrogens by both intrauterine tissues of 6-day pregnant rats. This metabolism could be locally modulated in IS through the participation of the estrogen sulfatase, the activity of which is in turn controlled by the presence of free estrogens, possibly synthesized and/or secreted by the embryo, which has been shown to inhibit the sulfohydrolase activity.

Comparative glycolytic metabolism of sperm from normal, asthenospermic, and oligoasthenospermic men.

The glycolysis of spermatozoa from normal, asthenospermic, and oligoasthen ospermic men was studied using a respirometry technique to measure glucose utilization by the production of 14CO2 from glucose 14C (U-L). Lactate and pyruvate were measured by a spectrophotometric method using DNA as reference. Human spermatozoa preferred glucose to fructose as the glycolytic substrate when concentrations of these hexoses did not exceed normal concentrations in the blood. Spermatozoa from oligoasthenospermic men produced an average of 3.5 times more 14CO2 (345, 457 dpm/mg DNA/hour) than did spermatozoa from asthenospermic (88,837 dpm/mg DNA/hour) and normal men (96,595 dpm/mg DNA/hour). They also formed four times more lactate (9.63 mumoles/mg DNA/hour) than spermatozoa from normal men (2.33 mumoles/mg DNA/hour) and 6.4 times more pyruvate (2.90 mumoles/mg DNA/hour compared to 0.45 mumoles/mg DNA/hour). Spermatozoa from asthenospermic men formed amounts of lactate (3.01 mumoles/mg DNA/hour) and pyruvate (0.63 mumoles/mg DNA/hour) similar to those produced by spermatozoa from normal men.

Metabolic changes in human spermatozoa related to capacitation.

PIP: Metabolic changes in human spermatozoa related to capacitation were investigated. Glycolysis, motility, oxygen uptake, survival time, and c hanges in tetracycline-binding capacity were studied in spermatozoa expo sed to the following possible capacitating agents: 1) human cervical mucus, 2) human serum, 3) hydroxalpinx fluid, and 4) follicular cyst fluid. Mucus and serum did not produce marked changes. Hydrosalpinx fluid produced an increase in oxygen uptake in the presence of citrate and succinate while follicular cyst fluid produced modifications that could be related to capacitation such as increase in oxygen uptake, increase in motility, and release of bound tetracycline. Similar changes were seen with cyclic adenosine monophosphate and adenosine triphosphate treatment but not with other related nucleotides. The parameters studied in this work are suggestive of functional capacitation.^ieng

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.

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.

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.

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.

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.

Inhibition of implantation by intraluminal administration of concanavalin A in mice.

Important characteristics of endometrial implantation sites are the changes of polarity and molecular composition of the cellular surface. For this reason the masking of surface carbohydrates could lead to an inhibition of the recognition by the blastocyst of the endometrial implantation site. Considering the impact of lectins on carbohydrates, we decided to utilize the intraluminal administration (5 microliter) of different concentrations of concanavalin A (15-60 microgram) in pregnant female mice in the preimplantation phase. An inhibition of 100% of implantation wqs obtained with concentrations of 30 and 60 microgram of the lectin administered on days 3 and 4 of the pregnancy (P less than 0.001). Less important effects were observed on administering 15 or 20 microgram of the lectin (73 and 87% of inhibition) and on utilizing the different doses on days 1 and 2 of the pregnancy. Thus, we conclude that the egg must recognize certain molecules of the endometrial surface (alpha-D-mannopyranose and alpha-D-glucopyranose) in order to implant and that the making of these sites potentially constitutes a new contraceptive approach.

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.

Uterine arginase inhibition affect the rat embryonic development.

The presence of polyamines (putrescine, spermidine and spermine) and the enzymatic activity of extrahepatic arginase (E.C. 3.5.3.1) which catalizes the hydrolysis of L-arginine into L-ornithine and urea have been related with cellular growth and development in several tissues. The enzymatic activity of arginase in rat implantation sites and its participation in reproductive process is demonstrated. Long-Evans adult rats during the 4th or 5th days of pregnancy were utilized. Arginase activity is higher in non-decidualized tissue (86.1 +/- 33 nmoles of urea/mg protein/min-1) when was compared with implantation sites (61.7 +/- 17). Intrauterine administration of several concentrations of a new synthetic L-ornithine analogue, AIAVA (2-amine-5-iodoacetamide valeric acid), produced embryonic growth arrest concomitant with arginase inhibition but not ornithine decarboxylase. From our results it is possible to stress the metabolic importance of uterine arginase in reproductive process.

Inhibition of rat embryonic development by the intrauterine administration of alpha-difluoromethylornithine.

Decarboxylation of L-ornithine by L-ornithine decarboxylase (ODC; E.C.4.1.1.17.) is the initial step in the biosynthesis of putrescine, ODC activity is generally low in most tissues, marked increases are associated with rapid tissue growth and particularly with mammalian embryogenesis. 0.5 mg/kg of DL-alpha-difluoromethylornithine (DFMO) (irreversible inhibitor of ODC) was administered to uterine horns of Long-Evans adult rats during the 4th day of pregnancy. As control material, saline (0.15 M) was administered to contralateral uterine horn. The animals were sacrificed on different days, the uterine horns were removed and the number of implant of implanted embryos were counted. DNA, RNA, protein and dry weight content in implantation sites (5th day of pregnancy) indicated that decidualization following DFMO took place normally but that embryonic growth was arrested in the treated horn. When 100 micrograms of putrescine were added together with DFMO, the embryotoxic effect was absent.

Inhibition of implantation by the intrauterine administration of phospholipases in the rat.

Phospholipases A2 and C (93 and 500 mU) were administered to uterine horns of Long-Evans adult rats during the first five days of pregnancy. As control material, saline (0.15 M) was administered to contralateral uterine horns. The animals were sacrificed on the ninth day of pregnancy, the uterine horns were removed and the number of implanted embryos were counted. Both horns were examined with light and electron microscopes. For electron microscopy, Ruthenium red was used to visualize possible changes of the outer coat (glycocalix) of the plasma membrane of endometrial epithelial cells. Implantation was inhibited when phospholipases A2 and C were administered during the first three days of pregnancy. Ultrastructural modifications included decrease of glycoproteins as demonstrated by diminution of the Ruthenium red staining that may indicate a decrease in the negative surface charges of endometrial surface epithelium.

Respiratory changes associated with the in vitro evagination of Taenia solium cysticerci.

The metabolic adaptability of Taenia solium cysticerci was studied in vitro, by measuring their respiratory rate before, during, and after trypsin-induced evagination. Under aerobic conditions, the oxygen consumption increased about 40% during evagination of the cysticeri and returned to basal rates after the process was completed. The percentage of evagination induced by trypsin was not affected under anaerobic conditions or in the presence of respiratory poisons such as cyanide and carbon monoxide. These data indicate that cysticerci use either aerobic or anaerobic pathways according to oxygen availability in the environment. Results from experiments of irreversible respiratory poisoning using cyanide suggest the presence of an alternative respiratory chain. Proteolytic action of trypsin on a fibrous layer surrounding the invaginated larvae is suggested by histological evidence.

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.