PubMed Health⌕ Search

Biomedical subjects

J C Lavoie

Publications and source records attributed to J C Lavoie.

At least 19 recordsLinked to original sources
PubMed1994

Antiperoxide activity of sodium metabisulfite. A double-edged sword.

Sulfites are chemical substances that are used widely in the pharmaceutical industry to reduce or prevent oxidation. Sodium metabisulfite (Na2S2O5) is still present in several parenteral amino acid solutions. Since intravenous lipid emulsions are contaminated by hydroperoxides, we evaluated whether metabisulfite had an antioxidant activity against hydroperoxides. In vitro, Na2S2O5 inhibited the oxidant activity of H2O2, tert-butyl-, and cumene hydroperoxides. The antioxidant capacity of metabisulfite was supported in vivo by the lower (P < 0.01) excretion of malondialdehyde, a stable end product of lipid peroxidation, in babies receiving metabisulfite in their parenteral nutrition. However, for concentrations outside the range found in solutions for parenteral nutrition, the reduction of hydroperoxides by Na2S2O5 could transform this compound into an oxidant, like a sulfite radical. It is suggested that metabisulfite has antiperoxide properties that, under specific conditions, contribute to the generation of toxic oxidants.

Antioxidants↗
PubMed1994

Gender-related response to a tert-butyl hydroperoxide-induced oxidation in human neonatal tissue.

Reports of gender-related differences in the activity of enzymes involved in the metabolism of intracellular antioxidants, led us to verify whether the prostaglandin response to tert-butyl hydroperoxide (TBH) differed according to the sex of infants. Segments of human umbilical veins were perfused in the presence or absence of TBH (0.25 mmol/l, and 1.0 mmol/l). Because TBH is quenched in the cell by glutathione peroxidase, total glutathione concentrations and production of glutathione-dependent prostaglandins (PGE2 and PGF2 alpha) as well as membrane-derived eicosanoids (PGI2 and thromboxane) were measured in the eluate. In veins from boys, TBH induced a sustained response for glutathione only, which was increased (p < 0.05). In female-derived tissue, the hydroperoxide induced a different response according to the dose of TBH. At 0.25 mmol/l, a drop (p < 0.005) in PGF2 alpha was associated with a rise (p < 0.001) in thromboxane. At 1.0 mmol/l, TBH had an opposite effect--there was a rise (p < 0.01) in PGE2 and PGI2. The prostaglandin concentration were not proportional to the oxidative stimulus, suggesting a critical level of TBH at which the oxidative state differs in tissues derived from boys or girls.

Dinoprost↗
PubMed1994

The increase in vasomotor tone induced by a parenteral lipid emulsion is linked to an inhibition of prostacyclin production.

The aim of the study was to verify whether the infusion of a lipid emulsion causes a rise in vascular pressure related to an imbalance in the production of vasoconstricting and vasodilatating eicosanoids. Segments of umbilical veins were perfused with and without 1.5 microM indomethacin (cyclooxygenase inhibitor) in solutions differing only in their lipid content (control vs. lipid). The lipid-induced higher pressure (p < 0.05) was associated with an inhibition (p < 0.05) in the output of the vasodilatator PGI2, and an increase (p < 0.01) in the production of the vasoconstrictor PGF2 alpha. Indomethacin abolished differences in pressure, but produced a rise (p < 0.01) in vascular tone of both the control and lipid-containing solutions by inhibiting PGI2 synthesis. Prostacyclin was the only eicosanoid significantly correlated (p < 0.01) to vascular tone. The lipid emulsion was therefore linked to the inhibition of the conversion of PGH2 to PGI2. The ensuing greater PGH2 availability would result in vivo, in the increased synthesis of vasoconstricting eicosanoids. The lipid-containing solution produced vasoactive responses similar to those reported with tert-butyl hydroperoxide, suggesting that hydroperoxides contaminating commonly used lipid emulsions could be causing a prostanoid-dependent vasoconstriction.

Cesarean Section↗
PubMed1993

Biologic effects of parenteral bisulfite on human vascular tissue.

Antioxidant properties of bisulfite are used to stabilize parenteral amino acid solutions. After reports of adverse reactions to dietary sulfites, we evaluated whether the infusion of bisulfite had biologic effects on human vascular tissue. Because an endothelial oxidative injury can affect mediators of vasoreactivity, vascular pressure and prostaglandin production were studied in an intact human vein model infused with clinically relevant amino acid solutions differing only by their metabisulfite content (0 versus 300 mg/L). The amino acid solution containing bisulfite presented higher venous pressure (p < 0.01) and prostaglandin production (6-keto-prostaglandin F1 alpha, p < 0.01; and prostaglandin E2, p < 0.05). A hydroxyl radical-generating system added to the solutions did not modify the pressure readings, but it resulted in an overall decrease in prostacyclin production (p < 0.05). Despite this known inhibitory effect on oxidative challenge on prostaglandin I2 production, prostaglandins remained higher in the presence of bisulfite. The results suggest that the effect of bisulfite takes place before prostaglandin H2 synthesis in the eicosanoid cascade, whereas the oxidative challenge affects specifically the synthesis of prostaglandin I2, after prostaglandin H2, indicating that there is no interaction between bisulfite and the hydroxyl generating system. Bisulfite has local vascular effects on endothelial mediators, separate from its antioxidant properties.

6-Ketoprostaglandin F1 alpha↗
PubMed1991

Total parenteral nutrition in the newborn infant: energy substrates and respiratory gas exchange.

The hypothesis that a high-fat parenteral regimen was beneficial for respiratory gas exchanges, in comparison with a high-glucose regimen, was tested in a paired crossover design. Ten parenterally fed newborn infants with no respiratory problems received two 5-day isoenergetic and isonitrogenous regimens that differed in their nonprotein source of energy; the level of fat intake (low fat (LF) 1 gm.kg-1.day-1; high fat (HF) 3 gm.kg-1.day-1) varied inversely with that of glucose. Continuous transcutaneous PO2 (tcPO2) and PCO2 (tcPCO2), respiratory gas exchange (indirect calorimetry), and plasma arachidonate metabolites were measured at the end of each regimen. Oxygen consumption and resting energy expenditure were not affected by modification of the source of energy. However, carbon dioxide production (VCO2) was higher during LF than during HF (6.9 +/- 0.2 vs 6.2 +/- 0.1 ml.kg-1.min-1; p less than 0.01), as was the respiratory quotient (1.08 +/- 0.02 vs 0.96 +/- 0.02; p less than 0.001). Despite the differences in VCO2, the tcPCO2 was not affected, suggesting adequate pulmonary compensation during LF, as documented by the higher minute ventilation (160 +/- 7 vs 142 +/- 5 ml.kg-1.min-1; p less than 0.01). The lower tcPO2 during the HF regimen (73.8 +/- 2.8 vs 68.8 +/- 2.6 mm Hg; p less than 0.015) indicated a disturbance at the alveolocapillary level induced by the lipid emulsion. No differences were found in circulating levels of prostaglandins and thromboxanes. The substitution of glucose for lipid did not modify fat storage (2.1 +/- 0.3 vs 2.1 +/- 0.3 gm.kg-1.day-1). We conclude that the supposed beneficial effect of a fat emulsion on respiratory gas exchange is questionable.

Blood Gas Monitoring, Transcutaneous↗
PubMed1990

Effects of glucocorticoids on prostaglandin formation by human amnion.

The human amnion may be an important source of prostaglandins involved in the onset of human labor and therefore it is important to define the factors that regulate their formation in this tissue. In the present study we demonstrate that glucocorticoids inhibit prostaglandin production by freshly isolated amnion cells. The inhibitory action of the glucocorticoids, however, changes to a stimulatory action when the cells are maintained in primary culture for a few days. For both inhibition and stimulation, concentrations of 10(-8) M dexamethasone or greater were required to give significant effects, and estradiol and progesterone had no effect on the prostaglandin output of the cells. Epidermal growth factor (EGF), which has previously been found to stimulate prostaglandin output by confluent amnion cells, did not alter prostaglandin output of cells initially placed in culture. Furthermore, the stimulatory action of EGF and dexamethasone appeared additive. The calcium ionophore A23187 stimulated prostaglandin output in freshly isolated cells and accentuated the inhibitory effect of dexamethasone. These studies indicate that prostaglandin formation by human amnion during pregnancy could be regulated by glucocorticoids. These steroids are easily available to the amnion by way of cortisone conversion to cortisol by the maternal decidua. The results also indicate that amnion is capable of responding to glucocorticoids in both a stimulatory and inhibitory fashion and whether one or both actions are of importance in vivo is a question that is as yet unresolved.

Amnion↗
PubMed1988

Cyclooxygenase products formed by primary cultures of cells from human chorion laeve: influence of steroids.

Cells were isolated from human chorion laeve obtained at term (38-40 weeks gestation) by elective caesarean section and were maintained in primary culture for 1 week in defined media supplemented with 10% fetal calf serum. The production of various cyclooxygenase products by the cultures was examined. Little or no prostaglandin (PG) F2 alpha, 6-keto-PGF1 alpha, thromboxane B2, or 13,14-dihydro-15-keto-PGF2 alpha was found. In contrast, the cells produced PGE2 which was low on day 0, increased during culture to a maximum on day 1 or 2, then declined to low levels. When cells were grown in the presence of media containing cortisol, dexamethasone, progesterone, and estradiol (at 10(-7) or 10(-9) M), the glucocorticoids (at 10(-7) and 10(-9) M), but not estrogen or progesterone, markedly inhibited the increase in PGE2 output. There was no difference in the protein content and thymidine incorporation of cells grown in the presence of glucocorticoids when compared with controls. This inhibitory effect was not sensitive to cycloheximide (1 microgram/mL) indicating protein synthesis may not be involved in the process. These studies indicate that PGE2 is the major prostaglandin formed by primary cultures of chorion laeve and that prostaglandin metabolism in the chorion is sensitive to glucocorticoid inhibition.

Cells, Cultured↗
PubMed1986

Steroid metabolism by cells from human chorion laeve isolated on Percoll gradients.

Collagenase-dispersed cells from human chorion laeve were examined on Percoll gradients. The 3 beta-hydroxysteroid dehydrogenase (a trophoblast marker) and steroid sulfatase activities of the cells were measured and a system was developed to isolate enriched preparations of the trophoblast cells. No cells were found to sediment at Percoll concentrations greater than 50%, and using continuous gradients of Percoll there appeared to be cells with different 3 beta-hydroxysteroid dehydrogenase (3 beta HSD): steroid sulfatase ratios sedimenting in different regions of the gradient. Cells with a high ratio were found in the denser region of the gradient. Continuous gradients provided inadequate separations of distinct populations of cells, thus to obtain a more reproducible system to isolate cells, discontinuous gradients of Percoll were studied. A discontinuous gradient composed of 5, 20, 40, and 60% Percoll was developed and three bands of cells were found sedimenting at the 20, 40 and 60% interfaces, respectively. The number and appearance of cells at the 20 and 60% interfaces varied from tissue to tissue. In contrast, the cells sedimenting at the 40% interface were less variable, a substantial number was found to be present in every tissue studied, they were similar in appearance to the trophoblast cells and had high 3 beta HSD:sulfatase ratios.

3-Hydroxysteroid Dehydrogenases↗
PubMed1986

Primary culture of cells from human chorion laeve: steroid metabolism and properties of cells grown in defined media supplemented with 0.1% or 10% fetal calf serum.

The purpose of this study was to develop primary cultures of human chorion laeve cells and examine certain aspects of steroid metabolism during culture. Tissues obtained by elective cesarean section at term (38-40 weeks) were dispersed with collagenase. Cells were isolated on Percoll gradients at the interface between 20% and 40% Percoll and examined in primary culture for up to 1 week. Cultures were carried out in chemically defined media supplemented with 10% or 0.1% fetal calf serum (FCS). The morphological and biochemical properties of the cells were different in the two systems. In 0.1% FCS, cells formed clumps of tissue within 16 h of plating, and there was no cell replication. In contrast, in 10% FCS, the cells formed a carpet of tissue and reached confluence after 5 days in culture, resulting in increased DNA and protein content and thymidine incorporation in the dishes. Three steroidogenic enzymes were studied during culture: alkyl steroid sulfatase, estrogen sulfatase and 3 beta-hydroxysteroid dehydrogenase. The sulfatases had higher activities in 0.1% than in 10% FCS, and their activities decreased markedly during the culture period. In contrast, 3 beta-hydroxysteroid dehydrogenase activity was higher in 10% FCS than in 0.1% FCS. Activity remained constant during the culture period in 0.1% FCS and increased in 10% FCS. In the latter system this increase resulted in the enzyme maintaining a constant specific activity during culture. These studies describe two viable systems of chorion laeve cells in primary culture, which may be valuable for studying long term and/or subtle effects on various metabolic aspects of this tissue.

3-Hydroxysteroid Dehydrogenases↗
PubMed1985

Steroid metabolism by human chorion laeve from dichorionic twin pregnancies.

Certain steroid metabolic properties of chorion laeve from dichorionic twin pregnancies were examined to determine whether they were present in chorion not contaminated by decidua or serum. In the chorion situated between the two amniotic sacs and not in contact with decidua, aryl sulfatase, 3 beta-hydroxysteroid dehydrogenase, and aromatase activities were found. This indicates that these reactions are present in chorion laeve and were not previously ascribed to this tissue because of decidual contamination. Specific cortisol binding was also present in this area of chorion laeve, which excludes serum contamination. It is suggested that the specific steroid-binding protein in the membranes may be derived from the transcortin-like protein present in amniotic fluid.

3-Hydroxysteroid Dehydrogenases↗
PubMed1985

Kinetic comparison of the 3 beta-hydroxysteroid dehydrogenase activity in human placenta, chorion laeve, and ovary.

Recent studies from our laboratory and others have shown that Km values for steroid substrates of the 3 beta-hydroxysteroid dehydrogenase in the human placenta were in the nanomolar range compared with micromolar values previously described. The purpose of the present study was to measure the kinetic parameters of the 3 beta-hydroxysteroid dehydrogenase in other human tissues, namely the ovary and chorion laeve, and to determine whether they were similar to those of the placental enzyme. In chorion laeve microsomes the 3 beta-hydroxysteroid dehydrogenase had Km values for dehydroepiandrosterone and pregnenolone similar to those found in placenta. Microsomes from human ovaries, on the other hand, had Km values for both substrates 10- to 20-fold higher. However, the ability of various steroids to inhibit the ovarian enzyme was similar to that previously described from the placenta and the chorion laeve.

3-Hydroxysteroid Dehydrogenases↗
PubMed1984

Kinetic studies on the formation of estrogens from dehydroepiandrosterone sulfate by human placental microsomes.

Using microsomes isolated from term human placentae kinetic analyses of each of the enzymes involved in estrogen synthesis from dehydroepiandrosterone sulfate have been carried out and the following parameters were found: sulfatase, Michaelis-Menten constant (Km) = 16,000 +/- 5,000 nM, maximum velocity (Vm) = 2.0 +/- 0.5 nmol X min-1 X mg protein-1; 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), Km = 15 +/- 3 nM, Vm = 1.8 +/- 0.4 nmol X min-1 X mg protein-1; aromatase, Km = 14 +/- 4 nM, Vm = 0.12 +/- 0.02 nmol X min-1 X mg protein-1. From these values one can predict that, theoretically, the rate-limiting enzyme in estrogen synthesis from dehydroepiandrosterone sulfate (DS) should change from the sulfatase at low concentrations of substrate to the aromatase at higher concentrations. In order to test this hypothesis we developed a system which allowed the formation of estrogens from DS, dehydroepiandrosterone, and androstenedione to be measured and the appropriate intermediates to be isolated. The sulfatase was found to be rate limiting at concentrations of DS below 2 microM and the aromatase was found to be rate limiting at higher concentrations. These data may explain why previous perfusion studies of human placentae indicated the sulfatase was the rate-limiting enzyme in estrogen synthesis yet in vitro studies found that it was the aromatase. Steroids previously shown to inhibit the 3 beta-HSD were examined for their ability to inhibit the formation of estrogens from DS. Although 3 beta-HSD activity was markedly inhibited this had little effect on the overall conversion of DS to estrogens, until high concentrations of inhibitors were used. The data also underline the importance of studying enzyme systems rather than single enzymes when studying steroid synthesis.

3-Hydroxysteroid Dehydrogenases↗
PubMed1980

In vitro conversion of pregnenolone to progesterone by term human fetal membranes.

The conversion of pregnenolone to progesterone by homogenates of term human fetal membranes (38 to 40 weeks' gestation) was quantitated prior to and after labor. Chorion pars reflexa obtained after spontaneous labor by vaginal delivery was more active than that obtained prior to labor by elective cesarean section (p less than 0.05). With homogenates of amnion pars reflexa (n = 18), and pars placentaris (n = 18) obtained prior to labor no conversion or pregnenolone to progesterone was detected. In contrast, in amnion pars reflexa obtained after labor the reaction was detected in five of the 13 tissues examined and was significantly different from tissues obtained prior to labor (p less than 0.01). However, the conversion by the amnion pars placentaris after labor was not significantly different from that to labor. Evidence for inhibition of the reaction in the amnion pars reflexa and pars placentaris obtained prior to labor was found when prolonged washing of these tissues (up to 5 hours) with isotonic KCl resulted in the conversion of pregnenolone to progesterone being easily detected. This study suggests that there may be "activation" of the conversion of pregnenolone to progesterone in fetal membranes during labor and indicates that considerable care should be taken in studying this and perhaps other reactions in fetal membranes as the mode of delivery, the preparation of tissue, and, in the case of the amnion, the area studied can affect the results obtained.

Amnion↗
PubMed1980

Substrate specificity of the placental microsomal aromatase.

Using an accurate and sensitive assay for the human placental aromatase we have found apparent Km values for androstenedione (4-androstene-3,17-dione) and testosterone to be 14 +/- 4.0 nM and 41 +/- 12 nM respectively. These values were significantly different (p < 0.001). Analyses at substrate concentrations 5-10 fold above and below the Km values did not indicate any anomalous kinetic behavior. Mixed substrate experiments were consistent with a single enzyme metabolizing both steroids: each competitively inhibited the aromatization of the other, and the "Ki" values were the same as their apparent Km values. Sodium chloride (1.2M) significantly increased the rate of testosterone aromatization by decreasing its Km value and had no significant effect on the aromatization of androstenedione. However, in the presence of this salt testosterone still inhibited the aromatization of androstenedione competitively with a "Ki" equal to its apparent Km. Our data is therefore consistent with the proposal that human placental microsomes contain a single "high affinity" site for the aromatization of androstenedione and testosterone.

Androstenedione↗
PubMed1978

3beta-Hydroxysteroid dehydrogenase activity in human fetal membranes.

A 3beta-hydroxysteroid dehydrogenase (3betaHSD) was demonstrated in term human fetal membranes (chorion and amnion) with both dehydroepiandrosterone (3beta-hydroxy-5-androsten-17-one) and pregnenolone (3beta-hydroxy-5-pregnen-20-one as substrates, and the subcellular distribution substrate and nucleotide specificity of the enzyme was studied. In both membranes the microsomal fraction (particles which sedimented at 105,000 g after 90 min) had the highest specific activity. The chorion was more active than the amnion but the enzyme in both tissues had similar substrate and nucleotide specificity. NAD was the preferred cofactor, and pregnenolone was a better substrate than dehydroepiandrosterone in the presence of NAD. However, with NADP as cofactor both steroids were equally good substrates. When the 3beta-hydroxysteroid dehydrogenase activity of chorion microsomes was compared with that of placental microsomes, the specific activities were found to be of the same order of magnitude, and the substrate, nucleotide specificity and steroid binding properties were almost identical.

3-Hydroxysteroid Dehydrogenases↗