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Biomedical subjects

R F Seamark

Publications and source records attributed to R F Seamark.

At least 145 records · Page 8Linked to original sources
PubMed1978

Lipid content of sheep ovarian follicles in culture.

A detailed examination was made of the lipid content of sheep ovarian follicles maintained in organ culture, and of the effect on this of human chorionic gonadotrophin. Lipid classes were isolated by thin-layer chromatography and the fatty acid composition determined by gas chromatographic analysis. The mean total lipid content of isolated follicles, 4-6 mm in diameter, after 18 h in organ culture was 43.1 nmol/mg wet weight tissue. Five major lipid fractions were detected, namely phospholipids (21.0 nmol/mg), cholesterol (14.5 nmol/mg), triglycerides (3.1 nmol/mg), free fatty acids (1.1 nmol/mg), and cholesteryl esters (3.4 nmol/mg). The phospholipid fraction was distinctive in that it contained a large percentage of sphingomyelin (28.0%). Other phospholipids present were phosphatidyl choline (42.0%), phosphatidyl inositol (6.9%), phosphatidyl serine (6.1%), and phosphatidyl ethanolamine and phosphatidic acid (16.9%). Each lipid fraction had a characteristic fatty acid profile which was rich in the content of unsaturated fatty acids. Inclusion of human chorionic gonadotrophin (20 i.u. ml-1) in the culture medium caused no discernable effects on the follicular content of lipids nor on the fatty acid composition of individual lipid fractions.

Animals↗
PubMed1978

Effects of human chorionic gonadotrophin on the release of cyclic AMP and progesterone from the sheep ovary at different stages of the oestrous cycle.

The rate of release of cyclic AMP by sheep ovaries containing a corpus luteum was determined at different stages of the cycle before and up to 60 min after an intra-arterial (ia) injection of 500 IU human chorionic gonadotrophin (hCG). The median cyclic AMP concentration in arterial plasma and of ovarian venous plasma following hCG stimulation was 93.2 and 98.0 pmol/ml, respectively. The ovaries of ewes examined at Days 1 and 2 of the cycle showed no response to hCG, whereas in 2 sheep at Day 3, hCG caused a slight response, and in 13 sheep examined between Days 5-18, hCG caused a marked increase in cAMP release. In 5 of the sheep in which both ovarian veins were cannulated, only the ovary with a corpus luteum responded to hCG with an increased secretion rate of cyclic AMP and progesterone. The results indicate a lack of responsiveness in the newly formed corpus luteum to hCG.

Animals↗
PubMed1978

Maturational changes in sheep ovarian follicles: gonadotrophic stimulation of cyclic AMP production by isolated theca and granulosa cells.

Theca and granulosa tissues isolated from sheep ovarian follicles of different sizes were incubated in the presence of human chorionic gonadotrophin (HCG; 5 IU/ml) or follicle stimulating hormone (FSH; 5 microgram NIH-FSH-S11/ml) for 40 min. Changes in the total amounts of cyclic 3',5'-adenosine monophosphate (cAMP) were used as an index of the responsiveness of these preparations to the hormones. Thecal tissue of both large (4-6 mm in diameter) and small (1-3 mm) follicles responded similarly to gonadotrophins. Granulosa cells from small follicles failed to respond to stimulation by HCG. FSH, however, consistently increased cAMP production in comparison with controls or cells treated with HCG. Granulosa cells of large follicles responded to both HCG and FSH.

Animals↗
PubMed1977

Metabolism of glucose, fructose and lactate in vivo in chronically cannulated foetuses and in suckling lambs.

1. Chronically cannulated sheep foetuses and suckling lambs were injected with 14C-labelled glucose, fructose or lactate, and sequential blood samples taken under conditions of minimal stress and without anaesthesia. 2. Gluconeogenesis from lactate was not detectable in foetal sheep, but the pathway was active in suckling lambs. 3. Fructose utilization rates were low in foetal sheep, with no measurable conversion into glucose or lactate. 4. The high rates of irreversible loss of both glucose and lactate in the foetus were decreased in suckling lambs. Radioactivity from labelled glucose entered both the lactate and fructose pools in foetal sheep, and entered the lactate pool in suckling lambs. 5. A model is proposed in which carbon flow between glucose, fructose and lactate has been quantified in foetal sheep.

Animals↗
PubMed1977

The appearance of gluconeogenesis at birth in sheep. Activation of the pathway associated with blood oxygenation.

1. Measurements of pyruvate carboxylase, mitochondrial phosphoenolpyruvate carboxykinase (GTP), hexose bisphosphatase and glucose 6-phosphatase in developing sheep liver showed substantial activities of all enzymes in the foetus, especially towards the end of gestation. Cytosol phosphoenolpyruvate carboxykinase (GTP) in livers of mid-term foetuses was only 10% of the activity at birth. 2. All enzymes except pyruvate carboxylase showed 1.5-2-fold increases after birth. 3. Gluconeogenesis form [14C]actate could not be detected in chronically cannulated sheep foetuses at any developmental stage and was not initiated by the infusion of adrenaline or glucagon. 4. An active pathway of gluconeogenesis was evident in vivo within 2 min after natural birth or within 4 min after Caesarian delivery of term lambs, and was delayed in prematurely delivered lambs until breathing was established and the blood fully oxygenated. 5. It is proposed that oxygen availability initiates gluconeogenesis in the newborn lamb.

Animals↗
PubMed1976

Oxygen uptake, glucose utilization, lactate release and adenine nucleotide content of sheep ovarian follicles in culture: effect of human chorionic gonadotrophin.

A study has been made of the oxygen uptake, glucose utilization, lactate release and cellular content of adenine nucleotides of isolated sheep ovarian follicles (4-6 min in diameter) maintained in organ culture, and of the effects on these parameters of the addition of human chorionic gonadotrophin (hCG). The mean oxygen consumption of the entire follicles when freshly isolated and of the theca and membrana components was 0-56, 1-08 and 0-05 mumol per milligram wet weight of tissue per hour respectively. About 8 mumol of glucose was taken up and 16 mumol of lactate released per milligram wet weight of follicular tissue per hour during the first 24-h period of culture. This rate reduced by about 30% for each subsequent day of culture, but was significantly increased by the addition of hCG. The mean ATP content of theca and granulosa tissues was 4-6 and 2-8 nmol/mg wet weight of tissue respectively. There was no discernable change in tissue adenine nucleotide content or energy charge ratio during the 3-day culture period, and prolonged exposure to hCG was without effect. Untreated follicles produced both oestrogen and androgens throughout the culture period. The addition of hCG resulted in a transitory stimulation in oestrogen secretion, inhibition of androgen secretion, and a marked and sustained rise in progestin secretion. It is proposed that the increase in glycolytic activity following exposure to hCG may relate to the activation of the granulosa cells coincident with a transference of steroid synthetic capacity from theca interna to membrana granulosa.

Adenine Nucleotides↗
PubMed1976

Changes in lymphocyte function during pregnancy.

A gradual increase in spontaneous lymphocyte DNA synthesis was demonstrated in each trimester of pregnancy. Autoradiographic studies indicated that lymphocytes were primarily responsible for this activity. PHA-induced lymphocyte transformation in both fetal calf serum and autologous serum was significantly reduced in the second and third trimesters of pregnancy. Spontaneous lymphocyte DNA synthesis was significantly reduced in patients with mild pre-eclampsia. However, no significant differences were seen in patients with severe pre-eclampsia in the third trimester of pregnancy compared with the normal control subjects. No evidence was adduced to implicate inhibitory humoral factors affecting the peripheral blood lymphocytes in pregnany patients in experiments in which washed lymphocytes were cultured in medium containing heterologous serum. In vitro experiments demonstrated that cortisol, progesterone and HPL caused a significant reduction in lymphocyte DNA synthesis, and HGH and HCG had a variable effect. However, only cortisol was regularly inhibitory at physiological concentrations. The progesterone effect was dose-related, producing 90 per cent inhibition of activity at a concentration of 10 mug/ml. No synergism could be shown between HPL and progesterone on lymphocyte transformation. The increase in activity of circulating immunoreactive cells during pregnancy and its depression with the onset of pre-eclampsia is discussed.

Chorionic Gonadotropin↗
PubMed1976

Cyclic AMP in sheep ovarian follicles: site of production and response to gonadotrophins.

This study was undertaken (i) to establish a relationship between cyclic AMP (cAMP) production and the degree of LH and FSH stimulation; (ii) to determine the effects of various gonadotrophins on follicular formation of cAMP; and (iii) to identify the precise intrafollicular site of cAMP formation. The formation of cAMP increased rapidly in follicles exposed to LH. Maximum concentrations were reached after 90 min and were maintained for 180 min. Extracellular release of cAMP increased steadily throughout the 180-min experimental period. Tissue levels of cAMP increased proportionally and significantly when LH concentrations in the medium were increased from 0 to 200 mi.u. ml-1. Tissue levels of cAMP were significantly increased by HCG, prostaglandin E-2 and noradrenaline, but not by prolactin, prostaglandin F-2alpha, serotonin or melatonin. Cyclic AMP formation occurred predominantly in the thecal compartment; the membrana granulosa contributed less than 3% of the total amount of cAMP formed after gonadotrophic stimulation. A significant amount of cAMP from the thecal cells was released into the extracellular compartment and appeared to pass into the granulosa cells.

Animals↗