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Biomedical subjects

D W Clarke

Publications and source records attributed to D W Clarke.

35 records · Page 2Linked to original sources
PubMed1986

Pharmacokinetics of ethanol and its metabolite, acetaldehyde, and fetolethality in the third-trimester pregnant guinea pig for oral administration of acute, multiple-dose ethanol.

The pharmacokinetics of ethanol and its metabolite, acetaldehyde, were determined in the third-trimester pregnant guinea pig (56-59 days gestation) for oral intubation of four doses of 1 g ethanol/kg maternal body weight, administered at 1-h intervals. Animals (n = 4-7) were sacrificed at each of selected times during the 26-h study. Ethanol and acetaldehyde concentrations were determined by headspace gas-liquid chromatography. The maternal and fetal blood ethanol concentration-time curves were virtually superimposable, which indicated unimpeded bidirectional placental transfer of ethanol in the maternal-fetal unit. The blood and brain ethanol concentrations were similar in each of the maternal and fetal compartments during the study, which indicated rapid equilibrium distribution of ethanol. There was accumulation of ethanol in the amniotic fluid resulting in higher ethanol concentration compared with maternal and fetal blood during the elimination phase, which indicated that the amniotic fluid may serve as a reservoir for ethanol in utero. Acetaldehyde was measurable in all the biological fluids and tissues at concentrations that were at least 1,000-fold less than the respective ethanol concentrations and were variable. There was ethanol-induced fetolethality that was delayed and variable among animals, and was 55% at 23 h. At this time interval, the ethanol concentrations in maternal blood and brain, fetal brain, and amniotic fluid were 35- to 53-fold greater and the acetaldehyde concentrations in maternal blood and fetal brain were four- to five-fold higher in the animals with dead fetuses compared with the guinea pigs with live litters. These data indicated that decreased ethanol elimination from the maternal-fetal unit was related temporally to the fetolethality.

Acetaldehyde↗
PubMed1986

Development of brain insulin receptors: structural and functional studies of insulin receptors from whole brain and primary cell cultures.

We studied the structural and functional characteristics of insulin receptors from rat brain and liver from late gestation through adulthood as well as from cultured neuronal and glial cells from neonatal rats. Specific insulin binding was present on membrane preparations from brain and liver at all stages of development studied, with maximal binding in neonates greater than 19-day-old fetuses greater than adults for both brain and liver. Maximal specific binding to cultured neuronal and glial cell membranes was similar (6.2% vs. 7.1%, respectively). [125I]Iodoinsulin cross-linking to the insulin receptor demonstrated that the mol wt (Mr) of the brain alpha-subunit was less than that of the liver alpha-subunit at all stages. [125I]Iodoinsulin cross-linking also demonstrated that the glial cell alpha-subunit (Mr, 130,000) migrated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis to a position intermediate between the liver (Mr, 135,000) and brain (Mr, 119,000), whereas the neuronal cell alpha-subunit (Mr, 118,000) comigrated with the brain alpha-subunit. In solubilized lectin-purified preparations from brain and liver during development as well as from neuronal and glial cells, insulin stimulated phosphorylation of the beta-subunit. The Mr of the brain beta-subunit, as determined by migration on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was less than that of the liver beta-subunit. The neuronal cell beta-subunit comigrated with the brain beta-subunit while the glial cell beta-subunit migrated to a position intermediate between the brain and liver beta-subunit. Solubilized lectin-purified preparations from all tissues demonstrated insulin-stimulable phosphorylation of exogenous substrates. From these studies we conclude that 1) functional insulin receptors are present in the brain during development in the rat; and 2) the structural differences demonstrated between neuronal and glial cell and between brain and nonneuronal insulin receptors taken together with previously demonstrated functional differences of the insulin receptor on these tissues suggest a unique function for insulin receptors on neuronal tissues.

Age Factors↗
PubMed1986

Insulin is a growth-promoter of the developing brain: possible implications for the infant of the diabetic mother.

The combination of diabetes mellitus and pregnancy was once associated with significantly increased morbidity and mortality for mother and baby. Advances in the obstetric management of the mother with diabetes and parallel advances in the management of the infant of the diabetic mother (IDM) have lowered the morbidity and mortality in many centers to near those for pregnancies in the non-diabetic woman. A significant increase still exists in the prevalence of congenital malformations in IDMs. Recent animal studies have shown that these malformations are related to maternal hyperglycemia early in the pregnancy, and that if it is controlled with insulin during the critical period of embryologic development, then the malformations may be prevented.

Animals↗
PubMed1985

Developing rat brain binds monoiodinated insulin isomers similarly to other extrahepatic target tissues.

We recently reported a series of binding and metabolic studies which led to the conclusion that the developing rat brain is a target tissue for insulin. Since insulin target tissues (extrahepatic) are capable of differentiating between various monoiodoinsulin isomers, we measured the binding of the B26 monoiodoinsulin isomer compared to the A14 in newborn rat brain preparations to determine if the developing rat brain shared the same relative binding of these isomers (viz. B26 greater than A14) with other extrahepatic tissues. The B26 isomer bound 1.57, 1.50 and 1.34 times as much as did the A14 to brain membranes, glia and neurons, respectively, whereas both isomers were bound equally by liver plasma membranes. Competition-inhibition curves were generated using homologous unlabeled (127I) insulin isomers. Binding of the B26 isomer was greater than the A14 at all concentrations. Scatchard plots showed that the receptor concentrations for the two isomers were similar, and affinity profiles showed that the differences in binding could be accounted for by the greater affinity of the receptors for the B26 isomer. The results indicate that the developing rat brain shares with other extrahepatic insulin target tissues a greater affinity for B26 monoiodoinsulin isomer compared to A14. Future studies of insulin binding should avoid using mixtures of iodinated insulins so that a uniform interpretation of data is made possible.

Animals↗
PubMed1985

Insulin receptors and insulin modulation of norepinephrine uptake in neuronal cultures from rat brain.

Neuronal cells from 1-day-old rat brain in primary culture have been utilized in the present study to characterize insulin-binding sites and a possible action of insulin on these cells. Binding of 125I-insulin to neuronal cultures was 90% specific and time-dependent and reached equilibrium in 120 min. Specific binding was reversible with greater than 90% of binding dissociable within 120 min with a t1/2 of dissociation of 15 min. Various insulin analogues competed for 125I-insulin binding to neuronal cultures according to their known biological potencies. Scatchard analysis of competition data yielded a typical curvilinear plot providing a class of high affinity (Kd = 11 nM) and low affinity (Kd = 65 nM) binding sites. Light microscopic autoradiographic analysis of 125I-insulin bound to neuronal cultures revealed the presence of silver grains predominantly on the neurites with occasional occurrence on the cell soma. Insulin had no effect on neuronal 2-deoxyglucose uptake in contrast with our previous findings demonstrating a 2-fold stimulation of 2-dGlc uptake into astrocyte glial cells from rat brain (Clarke, D.W., Boyd, F.T., Jr., Kappy, M.S., and Raizada, M. K. (1984) J. Biol. Chem. 259, 11672-11675). Incubation of neuronal cultures with insulin caused a dose-dependent inhibition of [3H]norepinephrine uptake with significant inhibition occurring at 1.67 X 10(-11) M. These findings demonstrate that: 1) neuronal cells in primary culture possess specific insulin receptors which are predominantly located on neurites and 2) insulin modulates monoamine uptake in these cultures which suggests that insulin may modulate neural signaling via specific neuronal insulin receptors.

Animals↗
PubMed1985

Disposition of ethanol in maternal blood, fetal blood, and amniotic fluid of third-trimester pregnant ewes.

The disposition of ethanol in maternal arterial blood, fetal arterial blood, and amniotic fluid of nine conscious, cannulated pregnant ewes (128 to 137 days' gestation) was determined for 1-hour maternal intravenous infusion of ethanol, 1 gm/kg maternal body weight. The maternal arterial blood and fetal arterial blood ethanol concentration-time curves were virtually superimposable up to 14 hours. The apparent zero-order ethanol elimination rates for maternal arterial blood and fetal arterial blood were similar. There was a time lag in the transfer of ethanol into amniotic fluid relative to fetal arterial blood, and the peak ethanol concentration in amniotic fluid was significantly lower than the concentrations in maternal arterial blood and fetal arterial blood. The apparent zero-order ethanol elimination rate for amniotic fluid was slower, but not significantly so, compared with the ethanol elimination rates for maternal arterial blood and fetal arterial blood. Ethanol-derived acetaldehyde was found in maternal arterial blood, fetal arterial blood, and amniotic fluid at concentrations at least 1000-fold lower than the respective ethanol concentrations. The data indicate that, for administration of this ethanol dosage regimen to the third-trimester pregnant ewe, there is rapid, bidirectional placental transfer of ethanol; elimination of ethanol from the fetus is regulated primarily by maternal elimination of ethanol; the amniotic fluid may serve as a reservoir for ethanol in utero; and there is appreciable acetaldehyde-metabolizing capacity.

Acetaldehyde↗
PubMed1985

Gas-liquid chromatographic analysis of ethanol and acetaldehyde in blood with minimal artifactual acetaldehyde formation.

A gas-liquid chromatographic procedure utilizing headspace gas analysis is described for the determination of ethanol and its metabolite, acetaldehyde, in a 100-microliters sample of blood from the rat, guinea pig, sheep, or human. Artifactual formation of ethanol-derived acetaldehyde is minimized during sample preparation by using a chemical solution containing perchloric acid and sodium azide in saline, and thiourea. Aqueous standards of ethanol and acetaldehyde are used to calibrate the procedure, and 1-propanol is used as the internal standard of the method. The recovery of ethanol and acetaldehyde from spiked blood samples is quantitative and reproducible, with a within-day coefficient of variation of less than 7% for ethanol and less than 9% for acetaldehyde. The lower limit of quantitative sensitivity is 0.006 mg/ml ethanol and 0.10 microgram/ml acetaldehyde. The instrumental analysis time is less than 3 min, which enables high sample throughput.

Acetaldehyde↗
PubMed1985

Differential pharmacokinetics for oral and intraperitoneal administration of ethanol to the pregnant guinea pig.

The disposition of ethanol was studied in third-trimester pregnant guinea pigs (56-59 days gestational age) following maternal administration of ethanol, 0.5 g/kg total body weight, by oral intubation and by intraperitoneal injection. For oral administration, exposure of the fetus to ethanol involved bidirectional placental transfer of ethanol between the maternal and fetal compartments. For ip administration, there was distribution of ethanol from the peritoneal space across the uterus and chorioamniotic membranes into the amniotic fluid in addition to absorption into the maternal blood circulation and subsequent placental transfer into the fetus. This resulted in exposure of the fetus to very high ethanol concentration in the amniotic fluid immediately following ethanol administration. The data indicate that the ip route of ethanol administration does not mimic ingestion of ethanol and should be avoided in future studies of the fetal alcohol syndrome in rodent animal models.

Administration, Oral↗
PubMed1985

Insulin stimulates macromolecular synthesis in cultured glial cells from rat brain.

The effect of insulin on macromolecular synthesis in glial cells cultured from brains of 1-day-old rats was studied to investigate the role of insulin in brain growth. Insulin caused a dose-dependent stimulation of protein synthesis (measured by [3H]valine incorporation into protein) that became significant by 7 nM insulin. Maximal stimulation of protein synthesis of 145% of control occurred with 18 nM insulin. Long-term protein synthesis was also stimulated to 136% of control by insulin in a dose-dependent manner after 6 days of insulin incubation. Insulin also stimulated net RNA and DNA synthesis (measured by [3H]uridine and [3H]thymidine incorporation into RNA or DNA, respectively) with significant stimulation by 2 nM insulin. Net RNA synthesis stimulation was maximal at 120% of control by 18 nM insulin. Plateau stimulation of DNA synthesis of 175% of control was reached by 200 nM insulin. The effects of insulin on glial protein and RNA synthesis appear to be mediated completely by the insulin receptor. Insulin, in physiological concentrations, stimulated glial DNA synthesis via its interaction with the insulin receptor (46% of total response). At supraphysiological concentrations insulin may have stimulated DNA synthesis via its cross-reactivity with the insulinlike growth factor I receptor (54% of total response). Thus insulin, at concentrations known to exist in the brain, stimulates the processes necessary for growth in the glial cell and is an important growth factor in the developing rat brain.

Animals↗
PubMed1984

Insulin binds to specific receptors and stimulates 2-deoxy-D-glucose uptake in cultured glial cells from rat brain.

The kinetics of 125I-insulin binding and physiological activity of insulin on glial cells cultured from brains of 1-day-old rats have been studied. Binding of 125I-insulin to cultured glial cells was specific, reversible, and time-dependent. Porcine and chicken insulin competed equally for 125I-insulin binding while other hormones or insulin analogs competed in proportion to their insulin-like biological activity. Incubation of glial cultures with insulin resulted in a time- and dose-dependent stimulation of 2-deoxy-D-glucose uptake. Maximal stimulation (190% of control) was observed with 18 nM insulin, and 0.1 nM insulin caused half-maximum effect. The stimulatory effect of insulin on 2-deoxy-D-glucose uptake was due to its effect on Vmax without affecting the Km. These observations suggest that insulin stimulates glucose uptake in glial cells cultured from rat brain, the effect mediated by insulin specific receptors.

Animals↗
PubMed1984

The disposition and the liver and thymus gland toxicity of 3,3',4,4'-tetrachlorobiphenyl in the female rat.

3,3',4,4'-Tetrachlorobiphenyl (TCBP) was administered orally to adult female Sprague--Dawley rats in the oral dosage regimen, 5 mg X kg-1 X day-1 for 21 days, followed by a 22-day postdosing period. Control animals received either the corn-oil vehicle (1 mL X kg-1 X day-1) or no treatment. 3,3',4,4'-TCBP distributed preferentially into the adipose tissue and liver, and apparent steady-state xenobiotic concentrations were attained in the adipose tissue (8 micrograms/g) and liver (300 ng/g) prior to the cessation of dosing. The 3,3',4,4'-TCBP concentrations in the serum, brain, kidneys, and thymus gland were lower and more variable than those in the adipose tissue and liver. During the postdosing period, 3,3',4,4'-TCBP was eliminated from the adipose tissue and liver by apparent first-order kinetics with elimination half-life values of 2.5 days and 0.8 day, respectively. The major route of excretion of unmetabolized 3,3',4,4'-TCBP was via the feces, and the amount excreted over 24 h did not exceed 8% of the dose administered on any given day. Throughout the experiment, there were no differences in the body weight or food and water intake for the 3,3',4,4'-TCBP-treated animals compared with the corn-oil-treated and nontreated rats. There was a significant increase in liver weight and a significant decrease in thymus gland weight for the 3,3',4,4'-TCBP-treated rats compared with the corn-oil-treated rats at the cessation of dosing and at 11 days thereafter, but there were no observable histological changes in these organs as assessed by light microscopy.(ABSTRACT TRUNCATED AT 250 WORDS)

Adipose Tissue↗
PubMed1983

Gas-liquid chromatographic determination of the distribution of 3,3',4,4'-tetrachlorobiphenyl in the adult female rat following short-term oral administration.

A gas-liquid chromatographic assay using electron-capture detection was developed for the quantitation of 3,3',4,4'-tetrachlorobiphenyl (TCBP) in the serum, urine, brain, liver, adipose tissue, and feces of the rat. The sample preparation involves extraction of 3,3',4,4'-TCBP with hexane under neutral or alkaline conditions (and washing with concentrated acid for feces only). Aqueous standards are used for calibration of the assay, except for adipose tissue. The lower limit of quantitative sensitivity of the assay for 3,3',4,4'-TCBP is 25 ng/mL for serum and urine and 125 ng/g for brain, liver, adipose tissue, and feces, which can be extended to 5 ng/mL and 25 ng/g, respectively, by analyzing a larger aliquot of the hexane extract. The overall accuracy is greater than 95% for serum, urine, brain and feces and 86% for liver, and the within-day coefficient of variation does not exceed 8.6%. 3,3',4,4'-TCBP was administered orally to adult, female, Sprague-Dawley rats in the dosage regimens: 0.2, 0.5 and 2 mg X kg-1 X day-1 for 10 days and 5 mg X kg-1 X day-1 for 4 days. 3,3',4,4'-TCBP distributed preferentially into adipose tissue and liver, where the xenobiotic concentration was greater in adipose tissue. The adipose tissue and hepatic 3,3',4,4'-TCBP concentrations were dependent on both the absolute dose and dosing schedule of the xenobiotic. Only trace concentrations, usually below the lower limit of quantitation, were detected in the serum, brain and kidney. Fecal excretion of 3,3',4,4'-TCBP was greater than urinary excretion for the 5 mg X kg-1 X day-1 X 4-day regimen.

Administration, Oral↗
PubMedDate not supplied

Accuracy of two systems for blood glucose monitoring without a meter (Chemstrip/Visidex).

Blood specimens collected from 159 campers were tested using Visidex (VD) (Ames Company, Elkhart, Indiana) and Chemstrip (CS) (Bio-Dynamics, Boehringer Mannheim, Indianapolis, Indiana) compared with glucose analyzer (GA) results. Readers were physicians, two with no prior experience using either strip and two with some experience using CS. Subsequently, 30 clinic patients and 4 staff contributed random specimens that were read by a trained novice as 68 discrete samples and compared with GA results. In both studies readers did not place blood on the strips and were thus blinded as to source and the possible previous reading with the other strip. Correlations of VD and CS with GA for camp and clinic data were calculated. Data were compared for reliability and errors greater than 20% of reference values (GA) were compared for VD and CS. CS correlated better with GA than did VD estimates. Reliability was also significantly greater for CS than for VD readings. Both of these visually read methods provided clinically useful estimates of glycemia.

Autoanalysis↗