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Biomedical subjects

G B Thomas

Publications and source records attributed to G B Thomas.

At least 55 records · Page 3Linked to original sources
PubMed1988

Secretion of prolactin in response to serotonin requires an intact hypothalamo-pituitary axis in the ewe.

The effects of prolactin secretion and the subsequent site of action of serotonin (5-HT) were examined in ovariectomized hypothalamo-pituitary intact (HP-intact) and ovariectomized hypothalamo-pituitary disconnected (HPD) ewes. The administration of 5 mg per animal of 5-HT increased plasma prolactin concentrations within 20 min by 2- to 3-fold in HP-intact ewes, but had no effect in HPD ewes. Injection of vehicle (saline) did not alter plasma prolactin concentrations in either group. These data indicate that 5-HT is involved in the control of prolactin secretion in sheep and that this action is exerted at the level of the hypothalamus and not directly at the anterior pituitary gland.

Animals↗
PubMed1988

Antimicrobial activity of amniotic fluid against Chlamydia trachomatis, Mycoplasma hominis, and Ureaplasma urealyticum.

Amniotic fluids obtained by amniocentesis at 16 weeks to term were examined for the presence of Chlamydia trachomatis, Mycoplasma hominis, and Ureaplasma urealyticum. Of 140 fluids tested, none harbored chlamydiae, and only one harbored mycoplasma, M. hominis. A number of amniotic fluids were subsequently tested for their ability to inhibit the growth of these microorganisms. Amniotic fluids and chlamydial suspensions in a 2:1 ratio were incubated 30 to 90 minutes before their inoculation in McCoy cells. Procedures were followed for chlamydial isolation. Genital mycoplasmas were incubated with amniotic fluid samples for 24 or 48 hours at 35 degrees C. Growth in amniotic fluid specimens was compared with growth in pseudoamniotic fluid and broth controls. Fourteen amniotic fluid specimens collected from gestations of 16 to 40 weeks, were found to be inhibitory to the formation of inclusions of C. trachomatis in McCoy cells. Ten amniotic fluid specimens (16 to 39 weeks, gestation) demonstrated various degrees of inhibition against M. hominis, and three fluids were inhibitory to the growth of Ureaplasma. The inhibitor was heat and protease resistant and activity was proportional to concentration. The molecular weight of the inhibitor was probably greater than 10,000 daltons, and pH, although perhaps a contributing factor, was not the cause of the inhibition.

Amniotic Fluid↗
PubMed1988

Rescue factor: a design for evaluating long-acting analgesics.

A design is described that uses need for supplemental (rescue) analgesic as a factor predicting effectiveness of a test analgesic. This methodology is especially suited for evaluating long-acting analgesics given repeatedly. Rescue use is measured over dosing intervals as test drug is titrated from a subanalgesic dose to that requiring no or minimal rescue. This design was used to evaluate oral long-acting morphine sulfate (MS Contin) given every 12 hours in a crossover study of cancer pain using oral immediate-release morphine sulfate given every 4 hours as reference. Less morphine was required for MS Contin given every 12 hours relative to immediate-release morphine sulfate given every 4 hours (186 +/- 22 mg vs. 239 +/- 35 mg; p = 0.04). Total daily morphine for both regimens correlated linearly (r = 0.96) with a slope of 1.27 +/- 0.11, significantly (p = 0.03) different from equivalence (slope of unity) in favor of MS Contin. This design features assay sensitivity (dose-response) and provides relative potency estimates for analgesics given at specific regimens.

Administration, Oral↗
PubMed1988

Secretion of LH, FSH and oestradiol-17 beta during the follicular phase of the oestrous cycle in the ewe.

Plasma concentrations of LH, FSH and oestradiol-17 beta were measured in blood samples taken at 15 min intervals for 48 h during the follicular phase of four Merino ewes. The amplitude of pulses of LH and the mean concentration of LH were higher at the beginning of the follicular phase, 36-24 h before the preovulatory surge of LH (amplitude 2.4 ng ml-1, mean concentration 3.9 ng ml-1), than at the end, 24-0 h before the preovulatory surge (amplitude 1.2 +/- 0.1 ng ml-1; mean concentration 1.4 +/- 0.1 ng ml-1). There was no change in the inter-pulse interval during this time (mean 74 +/- 5 min). Over the same period, oestradiol levels increased from 7-8 pg ml-1 to a peak of 10-15 pg ml-1. Mean FSH concentrations declined (36-24 h: 3.6 ng ml-1 vs 24-0 h: 1.8 +/- 0.3 ng ml-1) before rising at the time of the preovulatory surge of LH and again 24 h later. It was concluded that the biphasic response of LH to oestrogen that is seen in ovariectomized ewes may also operate during the follicular phase of the oestrous cycle in entire ewes.

Animals↗
PubMed1988

Effect and site of action of hypothalamic neuropeptides on prolactin release in sheep.

In an attempt to identify a physiological prolactin-releasing factor in the sheep, ovariectomized ewes were given intracarotid injections (10(-8)-10(-7) mol/animal) of thyrotropin-releasing hormone (TRH), vasoactive intestinal polypeptide (VIP), peptide histidine-isoleucine amide (PHI), oxytocin (OT), arginine vasopressin (AVP), substance P (SP), bombesin (BB), neurotensin (NT) and neuropeptide Y (NPY). Administration of TRH, AVP, NT and OT resulted in immediate and significant increases in plasma prolactin concentrations, the greatest stimulatory effect being obtained after TRH; other peptides had no effect in ovariectomized hypothalamo-pituitary intact ewes. AVP, NT and OT failed to release prolactin in ovariectomized ewes. These results suggest that (1) AVP, NT and OT may act via the hypothalamus to regulate prolactin secretion in hypothalamo-pituitary intact ewes; (2) VIP, PHI, SP, BB and NPY appear to have no direct roles at the pituitary level to control prolactin secretion in sheep, and (3) TRH stimulates prolactin secretion in ovariectomized ewes by a direct pituitary action.

Animals↗
PubMed1988

Gonadotropin-releasing hormone associated peptide (GAP) and putative processed GAP peptides do not release luteinizing hormone or follicle-stimulating hormone or inhibit prolactin secretion in the sheep.

A series of experiments was performed to monitor plasma luteinizing hormone (LH), follicle-stimulating hormone (FSH), and prolactin responses to human gonadotropin-releasing hormone (GnRH) associated peptide (GAP) and related peptides. Ovariectomized hypothalamo-pituitary disconnected (HPD) ewes were challenged with injections (1-10 micrograms i.v.) of GAP, or given, with and without estradiol, hourly 500- or 1,000-ng pulses of GAP for 5-7 days. In all cases GAP failed to cause the release of LH or FSH from the pituitary gland or to alter mean plasma prolactin concentrations. When the same HPD ewes were given hourly or 2-hourly pulses of 250 ng GnRH, LH responded in a a pulsatile manner, and FSH secretion was maintained, thus confirming the functional integrity of the pituitary gland after HPD. Fragments of the GAP molecule (pro-GnRH 14-36, 28-36, 38-49, and 51-66) and GAP dimer did not stimulate LH or FSH or inhibit prolactin release in HPD ewes. GAP and GAP dimer did not affect pituitary responsiveness to GnRH administration. GAP also failed to inhibit the thyrotropin-releasing hormone-induced rise in prolactin. Finally, GAP injections (100 micrograms i.v.) given to lactating ewes did not cause any change in plasma prolactin concentrations. These data show that human GAP, GAP dimer, or putative processed GAP peptides do not act on the sheep pituitary gland in a variety of physiological states to regulate gonadotropin or prolactin secretion.

Animals↗
PubMed1988

Effects of breed, ovarian steroids and season on the pulsatile secretion of LH in ovariectomized ewes.

The effects of season and of oestradiol and progesterone on the tonic secretion of LH were studied in ovariectomized Merino and Suffolk ewes, two breeds which differ markedly in the seasonal pattern of their reproductive activity. In the absence of exogenous steroids, the frequency of LH pulses was lower and the amplitude of the pulses was higher in anoestrus than in the breeding season for Merino and Suffolk ewes 30 days after ovariectomy. In long-term (190 days) ovariectomized ewes, this seasonal change in LH secretion was observed in Suffolk ewes only. During seasonal anoestrus, treatment of ewes with subcutaneous oestradiol-17 beta implants (3, 6 or 12 mm in length) decreased the frequency of LH pulses in a dose-dependent manner, with Suffolk ewes being far more sensitive to the inhibitory effects of oestradiol than Merino ewes. The lowest dose of oestradiol (3 mm) had no effect on the secretion of LH in Merino ewes, but reduced secretion in Suffolk ewes. Treatment of ewes with the highest dose of oestradiol (12 mm) completely abolished LH pulses in Suffolk ewes, whereas infrequent pulses remained evident in Merino ewes. During the breeding season, oestradiol alone had no effect on the pulsatile release of LH in either breed, but in combination with progesterone there was a significant reduction in LH pulse frequency. Progesterone effectively decreased LH secretion in both breeds in both seasons. It was concluded that differences between breeds in the 'depth' of anoestrus could be related to differences in the sensitivity of the hypothalamus to both negative feedback by oestradiol and the direct effects of photoperiod.

Animals↗
PubMed1988

Release of prolactin is independent of the secretion of thyrotrophin-releasing hormone into hypophysial portal blood of sheep.

In order to determine whether pituitary prolactin release was directly related to the secretion of TRH into hypophysial portal blood, serial portal and jugular venous blood samples were collected from seven lactating and three non-lactating ewes. In another experiment, samples were collected from five ovariectomized ewes while being exposed to an audio-visual stress and then later administered with chlorpromazine. Secretion of TRH was pulsatile in all ewes and independent of prolactin secretion; TRH pulses coincided with significant increases in prolactin secretion in only 15% of cases and only 29% of prolactin pulses were associated with TRH pulses. Sixty-seven per cent of suckling bouts were associated with increases in prolactin secretion, but only 22% of these were associated with significant increases in TRH secretion. Chlorpromazine increased prolactin levels fourfold but did not affect portal concentrations of TRH. Audio-visual stress was not a reliable method of causing prolactin release in this model. Mean portal concentrations of TRH and jugular concentrations of prolactin were not significantly correlated. These results show that hypothalamic TRH and pituitary prolactin are secreted independently in the sheep, implying that increases in prolactin release caused by suckling or chlorpromazine are not the direct result of increased TRH secretion.

Animals↗
PubMed1987

Secretion of prolactin in response to serotonin requires an intact hypothalamo-pituitary axis in the ewe.

The effects on prolactin secretion and the subsequent site of action of serotonin (5-HT) were examined in ovariectomized hypothalamo-pituitary intact (HP-intact) and ovariectomized hypothalamo-pituitary disconnected (HPD) ewes. The administration of 5 mg per animal of 5-HT increased plasma prolactin concentrations within 20 min by two-three fold in HP-intact ewes, but had no effect in HPD ewes. Injection of vehicle (saline) did not alter plasma prolactin concentrations in either group. These data indicate that 5-HT is involved in the control of prolactin secretion in sheep and that this action is exerted at the level of the hypothalamus and not directly at the pituitary gland.

Animals↗
PubMed1987

Pulsatile LH secretion during the preovulatory surge in the ewe: experimental observations and theoretical considerations.

The secretion of LH during the preovulatory surge has been shown to be pulsatile in several species. We confirmed this phenomenon in intact ewes in two experiments in which we studied the LH surge in detail. In the first experiment, the levels of LH were measured in discrete blood samples taken every 5 min around the onset of oestrus in 5 intact Merino ewes. For 2-5 h immediately prior to the onset of the surge, the median pulse interval was 17.5 min (range 10-55) and the median pulse amplitude was 0.70 ng/ml (range 0.28-1.33 ng/ml). During the ascending phase of the surge, the pulse interval did not change significantly (median 18.3 min) but the median pulse amplitude increased 20 fold to 18.0 ng/ml. During the descending phase, the median pulse interval was 22.5 min and the median amplitude decreased to 8.1 ng/ml (P less than 0.005). During the surge, intervals of 10 min and amplitudes over 30 ng/ml were common. In the second experiment with 2 oestrous Romanov x Préalpes-du-Sud ewes, blood was withdrawn continuously and samples were pooled over 4 min periods for 19-27 h. In the resulting profiles, individual pulses were better defined but the pulse frequencies (median interval 15.2 min during ascending phase) and amplitudes (median 35.0 ng/ml) varied in similar fashion to the Merinos. Using a simple mathematical description of pulsatile secretion, and assuming that LH is only released in pulses, we also found that the high concentrations normally achieved during the surge can only be explained by pulse frequencies and amplitudes well above those normally observed in other physiological situations, including the follicular phase or following ovariectomy. These findings strongly support the experimental data and the combined results of the two studies suggest that the initiation and execution of the LH surge is probably accomplished by changes in the amplitude of pulses. A high frequency is also essential and our data suggest that it may be established some time before the onset of the surge proper, but this requires confirmation. Respectively, the changes in amplitude and frequency reflect the pituitary and hypothalamic components of the positive feedback mechanism. The theoretical study suggested that a change in clearance rate could also contribute to the surge and this awaits further investigation.

Animals↗
PubMed1987

Effect of immunization against progesterone on oestrus, cycle length, ovulation rate, luteal regression and LH secretion in the ewe.

The effects of active immunization against progesterone on reproductive activity were studied in Merino ewes. Immunization against progesterone caused a shortening (P less than 0.01) of the interval between ovulations from 17-18 days (controls) to between 6 and 10 days (immunized group); this was associated with a corresponding reduction in the interval between LH surges. The immunized ewes also had higher (P less than 0.05) ovulation rates (1.72) than controls (1.25) and exhibited a reduced (P less than 0.01) incidence of oestrus (26% v. 95%). Many immunized ewes continued to ovulate despite the persistence of corpora lutea from earlier ovulations which led to an accumulation on the ovaries of many corpora lutea of different ages. The frequency of LH pulses in ewes immunized against progesterone (1.8 +/- 0.2 pulses/4 h) was significantly (P less than 0.001) higher than that of control ewes (0.3 +/- 0.1 pulses/4 h). This study highlights the importance of progesterone in the control of oestrus, ovulation, ovulation rate, luteal regression and the secretion of LH in the ewe.

Animals↗
PubMed1987

Steroid feedback inhibition of pulsatile secretion of gonadotropin-releasing hormone in the ewe.

The long-term negative feedback effects of sustained elevations in circulating estradiol and progesterone on the pulsatile secretion of gonadotropin-releasing hormone (GnRH) and luteinizing hormone (LH) were evaluated in the ewe following ovariectomy during the mid-late anestrous and early breeding seasons. GnRH secretion was monitored in serial samples of hypophyseal portal blood. Steroids were administered from the time of ovariectomy by s.c. Silastic implants, which maintained plasma concentrations of estradiol and progesterone at levels resembling those that circulate during the mid-luteal phase of the estrous cycle; control ewes did not receive steroidal replacement. Analysis of hormonal pulse patterns in serial samples during 6-h periods on Days 8-10 after ovariectomy disclosed discrete, concurrent pulses of GnRH in hypothalamo-hypophyseal portal blood and LH in peripheral blood of untreated ovariectomized ewes. These pulses occurred every 97 min on the average. Treatment with either estradiol or progesterone greatly diminished or abolished detectable pulsatile secretion of GnRH and LH, infrequent pulses being evident in only 3 of 19 steroid-treated ewes. No major seasonal difference was observed in GnRH or LH pulse patterns in any group of ewes. Our findings in the ovariectomized ewe provide direct support for the conclusion that the negative-feedback effects of estradiol and progesterone on gonadotropin secretion in the ewe include an action on the brain and a consequent inhibition of pulsatile GnRH secretion.

Anestrus↗
PubMed1987

GnRH secretion throughout the ovine estrous cycle.

In order to define the patterns of gonadotropin-releasing hormone (GnRH) secretion during the estrous cycle of the sheep we sampled hypophysial portal blood from conscious animals on day 1 of the cycle (n = 1), during the luteal phase (n = 8), during the follicular phase (n = 6) and during the preovulatory luteinizing hormone (LH) surge (n = 6). At the same time, we sampled jugular blood to measure plasma LH concentrations. During day 1 we noted regular GnRH pulses, whereas GnRH pulse amplitude and frequency were more variable in the luteal phase of the cycle. In the transition from the luteal phase to the follicular phase the GnRH pulse frequency increased and the amplitude decreased. Around the time of the LH surge we noted 3 types of secretory profiles for GnRH. In one sheep (type 1) there was a large GnRH pulse at the onset of the LH surge followed by very little activity during the surge. In two sheep (type 2) the GnRH profile did not change between the late follicular phase and the onset of the LH surge. In the remaining three sheep (type 3) there was a clear increase in the secretion of GnRH at the onset of the LH surge. With the exclusion of the type 1 sheep the GnRH pulse frequency was maximal (2 pulses/h) at the time of the LH surge; average portal GnRH levels were also maximal at this time.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

Differentiation of adipose tissue and muscle in hypophysectomized pig fetuses.

In the present study, fetuses were hypophysectomized (hypox) in utero on d 72 to 74 of gestation with an electrical cauterizing needle. One to six successfully hypox fetuses were removed on d 110 of gestation from each of five gilts. Subcutaneous adipose tissue samples and semitendinosus muscles were obtained from the hypox fetuses and an equal number of control fetuses. Body weights of control fetuses (n = 15; mean +/- SE, 1,195 +/- 33 g) were similar to weights of hypox fetuses (n = 15; 1,179 +/- 67 g). Fat cell size in the middle subcutaneous layer of adipose tissue was increased in hypox fetuses (P less than .01) compared with control fetuses. The number of obvious fat cell clusters (outer layer) in lipid stained sections was reduced (P less than .01) by 50% in hypox fetuses. Histochemical reactions for glucose-6-phosphate dehydrogenase, esterase and lipoprotein lipase (LPL) activities in middle layer cell clusters were considerably enhanced in sections from hypox fetuses compared with sections from controls. Quantitative analysis of percent light transmittance (Zeiss photometer) through LPL-stained cell clusters indicated an increase (P less than .001) in LPL staining in sections from hypox fetuses when compared with sections from control fetuses. Transverse muscle sections (cryostat) from hypox fetuses failed to show normal patterns (as seen in control muscles) of reactions for acid ATPase, malate dehydrogenase (NAD-dependent), NADH-TR and alpha-glycerol phosphate dehydrogenase (without NAD). The number of muscle fibers that were stained for these enzymes was greatly reduced in hypox fetuses compared with control fetuses. The number of lipid positive fibers was also reduced in hypox fetuses compared with control fetuses.(ABSTRACT TRUNCATED AT 250 WORDS)

Adipose Tissue↗
PubMed1987

Effect of bacterial growth on the bursting pressure of fetal membranes in vitro.

By mounting a layer of chorioamniotic membrane on a specially designed reaction vessel, we studied the effect of Escherichia coli and/or group B streptococcus growing on the decidual surface of the membranes in tissue culture or bacteriologic medium. The organisms grew equally well in either medium. When growing in tissue culture medium, either organism significantly weakened the membranes as compared with controls (membranes incubated in the absence of either organism). Membranes derived from pregnancies delivered vaginally or abdominally responded similarly. When organisms were grown in bacteriologic medium, bursting pressures did not decrease. Addition of bacteriologic medium (20-60%) to tissue culture medium did not affect bacterial growth, but inhibited significantly the lowering of bursting pressures. Bacteriologic medium also inhibited the peroxidase-H2O2-halide system in vitro. Heat-killed bacteria and/or supernatants of culture medium previously inoculated with bacteria were not effective in weakening membranes. The results suggest that live bacteria in conjunction with active membrane metabolism lead to a weakening and eventual rupture of the membranes.

Culture Techniques↗
PubMed1987

The development of perirenal fat depots in obese and lean pig fetuses.

The differentiation of perirenal adipose tissue was ascertained in fetuses from Ossabaw (wild and obese) and cross-bred lean dams at 70, 90 and 110 days gestation and from domestic lean and obese dams (USDA) at 110 days gestation. Condensation of presumptive adipocyte clusters was more apparent in Ossabaw sections than in cross-bred (lean) sections at 90 and 110 days gestation. Enzyme and lipid histochemistry showed fat cell cluster development to be largely independent of fetal strain (Ossabaw vs crossbred). However, adipocytes in Ossabaw fetuses were larger (110 days) and more reactive for esterase activity (all three ages) than were adipocytes from crossbred fetuses. Fetuses from USDA-obese dams had larger fat cell diameters (P less than 0.01), a higher percentage of lipoprotein lipase (LPL) positive cells than did fetuses from USDA-lean dams. Perirenal fat pad weights, percent depot lipid, percentages of glucose-6-phosphate dehydrogenase (G6PDH) and succinate dehydrogenase (SDH) positive cells were independent of fetal strain (USDA, lean vs obese). Fat cell cluster development was independent of fetal strain but large cellular condensations of presumptive cells were only present in sections from lean fetuses. Immature capillary beds (devoid of adipocytes) were demonstrable in lean sections but were not observed in obese sections. Morphological and histochemical results from both studies provide no evidence for brown adipocyte-like cells in fetal perirenal tissues.

Adipose Tissue↗
PubMed1986

Adipocyte development in primary rat cell cultures: a scanning electron microscopy study.

This study utilized scanning electron microscopy (SEM) techniques to observe primary cultures of stromal-vascular (SV) cells derived from postnatal rat inguinal adipose tissue. Cells were grown on collagen-coated, fibronectin-coated, or uncoated glass coverslips. Coverslips were normally fixed in glutaraldehyde, osmium tetroxide, dehydrated, and critical-point-dried. Other coverslips were frozen in isopentane (cooled in LN2) and dried or fixed in Baker's formalin for demonstration of inosine diphosphatase (IDPase) by X-ray microprobe analysis (XRMA). Adipocyte morphologies were similar on all substrates. At 2 days of culture, actin cables were detected extending from developing adipocytes. No difference in actin cable structure, cellular shape, or lipid accumulation was observed among the different substrates. Some stromal cells did not accumulate lipid but proliferated into a multilayer by 9 days in culture. Inosine diphosphatase was detected in the Golgi apparatus of developing adipocytes utilizing the technique of XRMA. This study demonstrates the potential for using SEM and XRMA techniques to define morphological features and cytochemical markers of adipocytes in vitro and the response of primary cultured rat SV cells to other attachment substrates.

Adipose Tissue↗