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Biomedical subjects

J M Bissonnette

Publications and source records attributed to J M Bissonnette.

At least 55 records · Page 3Linked to original sources

Effect of exercise on uterine blood flow in the pregnant Pygmy goat.

Pygmy goats in the last third of pregnancy were trained to walk on a treadmill at rates up to 2.0 mph and up an inclination of 0-15 degrees. Electromagnetic flowmeters were placed unilaterally on a uterine artery, and measurements were made while the goats were standing quietly on the treadmill and during 5 min of exercise. Blood flow fell during exercise in all five animals studied, and this reduction was proportional to the level of exertion. At the highest level of exercise that these animals would voluntarily perform, uterine artery blood flow fell by 32 +/- 3 (SE) % (P less than 0.001) from control. In four additional animals the radioactive microsphere technique was used to measure uterine blood flow at rest and after 5-7 min of exercise. In these animals, exercise caused total uterine blood flow to fall by 18 +/- 10%; cotyledonary (placental) blood flow fell by 8 +/- 13%, while myoendometrial blood flow decreased by 52 +/- 12% (P less than 0.05). Thus nonplacental portions of the pregnant Pygmy goat uterus suffer major reductions in blood flow during exercise. This vasoconstriction may be due to exercise per se or to concomitant hypocapnia or hyperthermia. Singleton and twin kids born to animals that exercised were of normal birth weight.

Animals↗

Effect of temperature on kinetics of hexose uptake by human placental plasma membrane vesicles.

Initial rates of passive and carrier-mediated D-galactose and D-glucose uptake were measured in membrane vesicles derived from the maternal surface of the human placental syncytiotrophoblast. Passive diffusion, as measured by L-glucose uptake, was slightly and continuously temperature-sensitive over a range 0-40 degrees C (Q10 = 1.1). Below approx. 26 degrees C, passive diffusion measured by D-galactose uptake in the presence of the inhibitor, cytochalasin B, was quantitatively similar to L-glucose uptake. Above this temperature, however, cytochalasin B appeared not to be as effective an inhibitor of carrier-mediated uptake. The initial rates of D-galactose carrier-mediated transport, generated at low concentration (10 microM) were very temperature-sensitive and yielded a non-linear Arrhenius plot. An Arrhenius plot of Vmax, generated with higher concentrations, was linear. The linearity of the Vmax Arrhenius plot, in conjunction with the high cholesterol content of this membrane preparation, suggests that a membrane lipid phase transition is not responsible for the non-linearity of the low concentration Arrhenius plot. A discontinuous temperature sensitivity of the interaction between D-galactose and the hexose transport system, as reflected by a marked sensitivity in Km, appears responsible for the non-linearity in this Arrhenius plot.

Carrier Proteins↗

D-Glucose-sensitive and -insensitive cytochalasin B binding proteins from microvillous plasma membranes of human placenta. Identification of the D-glucose transporter.

Cytochalasin B was found to bind to at least two distinct sites in human placental microvillous plasma membrane vesicles, one of which is likely to be intimately associated with the glucose transporter. These sites were distinguished by the specificity of agents able to displace bound cytochalasin B. [3H]Cytochalasin B was displaceable at one site by D-glucose but not by dihydrocytochalasin B; it was displaceable from the other by dihydrocytochalasin B but not by D-glucose. Some binding which could not be displaced by D-glucose + cytochalasin B binding site. Cytochalasin B can be photoincorporated into specific binding proteins by ultraviolet irradiation. D-Glucose specifically prevented such photoaffinity labeling of a microvillous protein component(s) of Mr = 60,000 +/- 2000 as determined by urea-sodium dodecyl sulfate acrylamide gel electrophoresis. This D-glucose-sensitive cytochalasin B binding site of the placenta is likely to be either the glucose transporter or be intimately associated with it. The molecular weight of the placental glucose transporter agrees well with the most widely accepted molecular weight for the human erythrocyte glucose transporter. Dihydrocytochalasin B prevented the photoincorporation of [3H]cytochalasin B into a polypeptide(s) of Mr = 53,000 +/- 2000. This component is probably not associated with placental glucose transport. This report presents the first identification of a sodium-independent glucose transporter from a normal human tissue other than the erythrocyte. It also presents the first molecular weight identification of a human glucose-insensitive high-affinity cytochalasin B binding protein.

Affinity Labels↗

Central chemical regulation of breathing movements in fetal lambs.

In 12 chronically prepared fetal lambs between 126 and 136 days of gestation, 17 ventriculocisternal perfusions (123 microliter/min) were performed. The concentration of bicarbonate ([HCO3-]) in the mock cerebrospinal fluid (CSF) perfusate was altered to manipulate the [HCO3-] in the cerebral ventricles. These perfusions did not systematically alter fetal arterial PCO2, PO2, pH, heart rate or mean blood pressure. Fetal breathing movements (FBM) were noted as rhythmic negative intrathoracic pressures with reference to amniotic fluid pressure. The incidence of FBM was determined and expressed as the percent of the 3 h observation period during which breathing movements were present. Perfusions with mock CSF having an approximately normal [HCO3-], resulted in cisternal [HCO3-] of 23.9 +/- 0.8 (SEM) meq/L and an incidence of fetal breathing movements (FBM) of 22.1 +/- 10.0%. Perfusions with an elevated [HCO3-] in the mock CSF increased the cisternal [HCO3-] to 30.7 +/- 0.4 meq/L and lowered the incidence of FBM to 0.6 +/- 0.2%. Perfusions with a lowered [HCO3-] mock CSF decreased the cisternal [HCO3-] to 17.3 +/- 0.8 meq/L and increased the incidence of FBM to 45.7 +/- 6.8%. These perfusions did not alter the relative incidence of low voltage (LV) versus high voltage (HV) electrocortical (ECOG) state. In 4 animals, low [HCO3-] perfusions induced FBM in the normally apneic HV ECOG state. We conclude that in the near-term fetal lamb, central chemoreceptors can modulate the incidence of rhythmic respiratory efforts, that tonic afferent stimuli arising from these receptors are critical for the generation of spontaneous FBMs and that central acidosis is capable of evoking FBMs in the normally apneic HV ECOG state.

Animals↗

Cerebral metabolism in hypoglycemic and hyperglycemic fetal lambs.

To investigate the mechanism whereby glucose affects fetal breathing movements (FBM), cerebral metabolism was studied in 12 unanesthetized fetal lambs, during fasting-induced hypoglycemia and after a subsequent fetal infusion of glucose. Preductal arterial and sagittal sinus blood samples were analyzed for glucose and oxygen concentrations and for blood gases and pH. Measurements of regional brain blood flow were made with radioactive microspheres. Maternal fasting of 24- to 36-h duration resulted in a decrease in fetal blood glucose from 1.061 +/- 0.085 mmol X l-1 to 0.664 +/- 0.053 (P less than 0.001). Although cerebral glucose and O2 uptake remained unchanged, sagittal sinus CO2 partial pressure (Pco2) and [H+] were significantly decreased and may have contributed to the observed decrease in FBM. A 2-h infusion of glucose to the fetuses of fasted animals resulted in an increase in blood glucose to 2.452 +/- 0.173 mmol X l-1. Cerebral O2 consumption was again unchanged; however, cerebral glucose consumption was significantly increased as were sagittal sinus Pco2 and [H+], which may have contributed to the observed increase in FBM. These findings are consistent with the hypothesis that glucose affects the incidence of FBM in part by altering the environment of central chemoreceptors.

Animals↗

Reconstitution of D-glucose transporter from human placental microvillous plasma membranes.

Proteins from microvillous plasma membrane vesicles of the maternal surface of human trophoblast were solubilized with octyl beta-D-glucoside. After incorporation of the soluble protein into phospholipid liposomes D-glucose uptake exceeded that of L-glucose. The reconstituted system showed that D-glucose uptake was not sodium dependent and was inhibited by cytochalasin B. Efflux of D-glucose from the proteoliposomes was retarded by cytochalasin B. D-Glucose uptake, but not L-glucose, was proportional to the amount of protein used in the reconstitution procedure. Membrane protein was also solubilized with octylglucoside from vesicles that had been extracted previously by dimethylmaleic anhydride. Proteoliposomes prepared from these latter proteins showed D-glucose uptake threefold greater than that from octylglucoside solubilization alone, but sodium dodecyl sulfate polyacrylamide gel electrophoresis of the extracted protein showed no clear difference between the double extraction procedure and the pattern obtained with the single detergent.

Carrier Proteins↗

Glucose uptake into plasma membrane vesicles from the maternal surface of human placenta.

Glucose uptake into plasma membrane vesicles from the maternal surface of the human placenta was measured with the Millipore filtration technique. Uptake off D-glucose was dependent on the osmolarity of the incubation medium surrounding the vesicles. Uptake of D-glucose exceeded that of L-glucose. The uptake of D-glucose was not enhanced by placing 100 mM NaCl or NaSCN in the medium outside the vesicles (none inside) at the onset of uptake determinations. D-glucose transport was inhibited by cytochalasin B; phloretin, phlorizin, and 1-fluoro-2,4-dinitrobenzene. D-glucose uptake was inhibited by 2-deoxy-D-glucose, 3-O-methyl-D-glucose and to a lesser extent by D-galactose. It was not inhibited by alpha-methyl-D-glucoside. Cytochalasin B binding to the vesicles was 30% inhibited in the presence of 80 mM D-glucose. The results indicate that the system for facilitated transport of D-glucose at the maternal face of the placenta is distinctly different from that on the brush-border membrane of intestine or renal tubule and more closely resembles that of human erythrocyte.

Cell Membrane↗

Central shunt flows and pressures in the mature fetal lamb.

Two to eight days after surgery, we determined biventricular cardiac output with the microsphere-dilution method and individual ventricular output with an electromagnetic flow sensor in 12 fetal lambs. Mean biventricular output was 462 ml.min-1.kg-1. Right ventricular output was 60%, pulmonary blood flow was 6%, blood flow in the ductus arteriosus was 54%, flow through the foramen ovale was 34%, and flow in the descending thoracic aorta above the ductus arteriosus was 12% of the biventricular cardiac output. Foramen ovale flow did not correlate with the pressure difference between the atria. The flow though the foramen ovale correlated positively with the flow in the inferior caval vein. It was concluded the shunt flow through the foramen ovale is mainly determined by the kinetic energy of the blood in the "lower atrial inflow tract." Fetal systemic somatic blood flow showed a reliable negative correlation with the oxygen tension in the fetal arterial blood. This was taken as presumptive evidence for long-term autoregulation of flow in the peripheral tissues.

Animals↗

Ventriculocisternal cerebrospinal perfusion in unanesthetized fetal lambs.

Methods have been developed for the placement of cannulas in the lateral cerebral ventricle and cisternum magnum of the fetal lamb. Three to seven days after surgery, perfusion of the cerebral ventricular system was carried out in unanesthetized lambs in utero. Artificial cerebrospinal fluid (CSF) was perfused at 123 microliter/min for 4-5 h with no change in fetal arterial O2 pressure, CO2 pressure, pH, mean arterial blood pressure, or heart rate. The preparation is suitable for measurements of net formation of fetal central CSF and also for producing changes in the ionic composition of the fetal CSF.

Animals↗

Placental transport of carbohydrates.

Glucose transport across the placenta takes place by means of a facilitated process. The transporter shows considerable specificity with respect to different hexoses. Transport across the microvillous plasma membrane of the trophoblast surface facing the intervillous space has been characterized. The details of transport across the fetal directed plasma membrane and the regulation of D-glucose membrane protein transporters remain unknown.

Animals↗

Effect of oxygen and of carbon dioxide tension on the incidence of apnea in fetal lambs.

Fetal breathing movements (FBM) in unanesthetized lambs in utero were correlated with measurements of arterial blood gases. One hundred and eighty-seven observations, consisting of the incidence of FBM during the hour preceding and the hour after the blood gas determinations, were made on 125 separate days in 30 fetuses of 117 to 146 days' gestational age. Fifty-eight percent of the observations with fetal apnea (FBM 0 to 9% in 2 hours) showed hypoxia (PaO2 less than or equal to 16 torr), whereas only 4% of the observations with FBM greater than 10% were associated with hypoxia. Sevety-eight percent of the hypoxic and normocarbic (PaCO2 42 to 53 torr) observations demonstrated apnea. However, only 44% of hypoxic plus hypercarbic (PaCO2 57 to 63 torr) fetuses were apneic, and with an elevated PaCO2, apnea tended to develop at a lower level of PaO2. We conclude that FBM may persist in the presence of hypercarbia with hypoxia.

Animals↗

Uterine oxygen uptake in the pregnant Pygmy goat.

The diffusion equilibrium method was used to measure uterine blood flow (Qut) and oxygen consumption (VutO2) of 7 Pygmy goats in the last third of pregnancy. VutO2 was 9.3 +/- 1.3 (mean +/- SEM) ml.min-1.kg-1 weight of the uterus and its contents: the placenta, fetus and fetal membranes. This O2 requirement was provided by a high blood flow (349 +/- 48 ml.min-1.kg-1) and a narrow arteriovenous difference (2.4 +/- 0.1 vol%) across the uterus. Maternal arterial O2 content (CaO2) was varied by incurring chronic anemia in 3 animals. CaO2 over the range 6.5 - 12.8 vol% did not affect uterine blood flow or the arteriovenous oxygen content difference across the uterus. Weight specific (ml.min-1.kg-1) VutO2 did not differ in animals whose total uterine weights varied over a wide range and uterine vein oxygen tension and content did not fall prior to delivery. We conclude that the absolute uterine blood flow varies over a wide range to meet the requirements of the uterine contents.

Animals↗

Postnatal regulation of canine oxygen delivery: erythrocyte components affecting Hb function.

A rightward shift in the blood oxygen dissociation curve occurs during the 1st mo of canine life. A detailed peptide analysis indicated that dogs do not have a separate fetal hemoglobin. Other erythrocyte components such as ATP, K+, Na+, and H+ were excluded as significant mediators of the postnatal oxygen affinity change. Erythrocyte 2,3-DPG levels essentially zero in fetal dogs, increased rapidly during the 1st mo of canine life. There was a significant correlation between this postnatal 2,3-DPG increase and the postnatal decrease in blood oxygen affinity. Dialyzed hemolysates of fetal or adult canine blood have the same intrinsic oxygen affinity and the same response to normal adult levels of 2,3-DPG. Furthermore, the magnitude and direction of this 2,3-DPG-induced decrease in oxygen affinity in vitro are comparable to the in vivo postnatal change in oxygen affinity.

Aging↗

The role of a trial of labor with a positive contraction stress test.

During a 28 month period 812 patients underwent antepartum FHR testing. Twenty-eight patients had a positive CST. There were two antepartum fetal deaths and 11 patients had a cesarean section without a trial of labor. Fifteen patients had a trial of closely monitored labor (continuous FHR and fetal scalp blood sampling when indicated) and 11 of these (73%) were delivered vaginally. The CST records were examined for: per cent late deceleration, baseline FHR, presence of FHR accelerations, duration of the latency period (time from onset of contraction to onset of deceleration), and amplitude of deceleration. The absence of accelerations (nonreactive CST) and a latency period of less than 45 seconds both predicted persistent late deceleration during labor or fetal death in utero but statistical significance was found only for the latter parameter.

Apgar Score↗

Hen's eggs with retarded gas exchange. I. Chorioallantoic capillary growth.

The diffusing capacity of the hen's egg for carbon monoxide (D tau co) increases during incubation, reflecting the development of the chorioallantoic circulation. Previous work showed that the increase in D tau co could be diminished by incubating eggs in a 60% oxygen environment. The present work explored the effects of impeding gas exchange on the development of D tau co. Half the shell area was covered during incubation by a removable neoprene membrane which limited D tau co by approximately 20%. No difference could be detected between the D tau co values (measured with the membrane removed) of these eggs and control eggs incubated in the same 21% oxygen environment without a neoprene membrane. We conclude that the development of the chorioallantoic circulation is at its maximum under normal conditions of incubation and cannot be accelerated by restricting gas exchange.

Allantois↗

Placental transfer of water and nonelectrolytes during a single circulatory passage.

A bolus of a 14C-labeled nonelectrolyte and tritiated water was injected into the maternal arterial system supplying the uterine circulation. Blood was sampled immediately on the fetal side of the placenta from the umbilical vein. A placental transfer index (PTI) was calculated from the ratio of the tracers in the fetal blood divided by the ratio of the tracers in the injectate. Placental transfer was measured for 11 nonelectrolytes in anesthetized guinea pigs and 8 nonelectrolytes in unanesthetized sheep. Placental transfer in both species was determined by molecular weight and lipid solubility. The PTI measured for the lipophilic solutes antipyrine and 1,6-hexanediol were less than 1, and those for methanol and ethanol were greater than 1. We conclude that placental transfer for water and small (mol wt less than 200) lipid-soluble nonelectrolytes is limited by diffusion as well as by blood flow.

Animals↗