bluuebunny/crossref_metadata_2025_split huggingface dataset:
Studies on the subcellular mechanisms mediating the negative estradiol effect on GnRH-induced LH-release by rat pituitary cells in culture
Authors: Emons Günter , Frevert Ernst U. , Ortmann Olaf , Fingscheidt Ulrich , Sturm Rita , Kiesel Ludwig , Knuppen Rudolf 
Published: December 2008

Abstract.
Cultured pituitary cells from adult female rats were treated for 4 or 24 h in the absence or presence of E2 (10−9 mol/1) with increasing concentrations of the transcription inhibitor actinomycin-D or the translation inhibitor puromycin. During the last 4 h of incubation, LH release was stimulated with 5 × 10−10 mol/1 GnRH. The positive E2 effect observed after 24 h treatment with the steroid was clearly abolished by actinomycin-D at concentrations ≥10−10 mol/1 and by puromycin at concentrations ≥ 10−5 mol/1. These findings indicate that the positive E2 effect on GnRH-induced LH release is fully dependent on intact mRNA and protein synthesis. The negative E2 effect observed after 4 h treatment with the steroid was not affected by actinomycin-D and abolished by puromycin only at concentrations ≥ 10−4 mol/1. Similar results were obtained, when cells had been treated for 4 h with actinomycin-D or puromycin before the 4 h E2 treatment started. Thus, the negative E2 effect seems to be independent of mRNA synthesis and dependent on protein synthesis to a lesser extent than the positive E2 effect. In an attempt to identify positively the subcellular mechanism via which E2 exerts its negative effect, several steps in the GnRH stimulus secretion coupling mechanism were checked whether or not they are modulated by E2. The negative effects of E2 (10−9 mol/1) on LH release induced by GnRH (10−10, 10−9 mol/1) and by the activators of voltage dependent Ca2+ channels K+ (64 mmol/1) or veratridine (3.3 × 10−5 mol/1) were comparable to those of the calcium antagonist verapamil (10−6 mol/1). These findings supported the speculation that E2 might act on the Ca2+ channels. The LH release induced by the Ca2+ ionophores A 23 187 (10 −4 mol/1) or ionomycin (6.6 × 10−5 mol/1), however, was also significantly reduced by 10−9 mol/1 E2, indicating that the steroid modulated a mechanism secondary to the increase of intracellular Ca2+. Also GnRH (10−9, 10−8 mol/1) induced accumulation of [3H]inositol phosphates was not influenced by E2 (10−9 mol/1) treatment, though the steroid exerted a significant negative effect on the LH release by these cells, indicating that phosphatidylinositol-4,5-biphosphate breakdown is not the point of attack for the estrogen. The LH release induced by the activation of protein kinase C with the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA, 10−7 mol/1) or with the synthetic diacylglycerol 1,2-dioleyl-sn-glycerol (DOG, 5 × 10−5 mol/1, 10−4 mol/1) was significantly reduced by E2 treatment. These findings give rise to the speculation that E2 might exert its negative effect on GnRH-induced LH release in cultured pituitary cells at the level of protein kinase C or a mechanism secondary to the activation of this enzyme.
Effect of estradiol and tamoxifen on GnRH self-priming in perifused rat adenohypophysial cells
Authors: Cover Patricia O. , Buckingham Julia C. 
Published: December 2008

Abstract.
Columns of enzymatically dispersed rat pituitary cells have been used to examine the effects of estradiol and the estrogen receptor antagonist, tamoxifen, on the responsiveness of the gonadotropes to repeated challenges with GnRH. Synthetic GnRH stimulated readily the release of LH from the pituitary cells. When challenged repeatedly with the same submaximal dose (8 nmol/1) of the releasing hormone, cells from animals in diestrus 2 or pro-estrus, like those from juvenile rats, showed a marked progressive increase in their sensitivity, but those from rats in estrus or diestrus 1 did not. Adenohypophysial cells removed from adult rats ovariectomized 14 days previously also failed to exhibit sensitization even when estradiol-17β (1 and 10 nmol/1) was included in the perifusion fluid. However, those from ovariectomized rats treated with appropriate doses of the steroid (1.5 μg/day sc for 14 days or 5 μg sc 22 h before decapitation) did. The capacity of pituitary cells from intact rats to exhibit priming on the anticipated day of pro-estrus was unaffected by the administration of the estrogen receptor antagonist, tamoxifen (700 μg/100 g sc on the morning of diestrus 1 and 24 h later). Similarly, addition of tamoxifen (1 μmol/1) to the incubation medium failed to influence the capacity of pituitary cells from juvenile rats to exhibit priming in vitro. The results support the concept that the ability of GnRH to 'prime' the gonadotropes is dependent on estradiol and indicate that the exposure of the hypothalamo-pituitary complex to critical levels of estradiol prior to autopsy is necessary for the gonadotropes to retain their capacity to exhibit priming in vitro.
Studies on the prostaglandin E-2-9-ketoreductase mediated production of prostaglandin F-2α and its metabolism in cultured rabbit luteal cells
Authors: Schlegel Werner , Daniels Dieter 
Published: December 2008

Abstract.
Luteal cells were isolated from pseudopregnant rabbits on days 3, 6, 9, 12 and 15 post ovulation. Prostaglandin concentration and the activities of the enzymes prostaglandin E-2-9-ketoreductase and prostaglandin-15-hydroxydehydrogenase were determined. Luteal cells from day 7 and 12 of pseudopregnancy were maintained in culture for 24 h and then exposed to a mixture of [1-14C]PGE2 (70 μmol/l) in the presence or absence of estradiol-17β. After a 24 h incubation period, the culture medium was adjusted to pH 3.5, immediately extracted and analysed for PG. Cultured luteal cells were able to convert exogenously applied PGE2 to PGF2α and to metabolize both PGs. Primary PGs as well as their metabolites 15-keto PGE2, 15-keto PGF2α, 13,14 dihydro15-keto PGE2, and 13,14 dihydro-15-keto PGF2α were detected in the culture medium and in the cells. The addition of estradiol-17β together with PGE2 caused a significant reduction in the PGE2-9-ketoreductase mediated PGF2α synthesis, whereas the metabolism of PGE2 remained unchanged. This inhibitory effect of estradiol-17β was dose-dependent on day 12 of pseudopregnancy. The results demonstrate that isolated rabbit luteal cells are able to synthesize and metabolize the luteolytic factor PGF2α. Whether the inhibitory effect of estradiol-17β may have any physiological relevance has to be examined in further studies.
Bovine neurophysin-II stimulates prolactin release without involvement of dopaminergic prolactin-release inhibiting factor receptor in the estradiol-primed male rat
Authors: Shin S. H. , Obonsawin M. C. , Stirling R. 
Published: December 2008

Abstract.
Neurophysins have been considered to be physiologically inert carrier proteins for the neurohypophysial hormones, oxytocin and vasopressin. We have observed that bovine neurophysin-II indirectly stimulates prolactin release in estradiol-primed male rats. The release of prolactin is regulated by a dual hypothalamic control system, the prolactin-release-inhibiting factor and the prolactin-releasing factor. We have tried to clarify whether neurophysin-II is acting through stimulation of prolactin-releasing factor by eliminating the possibility of dopaminergic prolactin release-inhibiting factor release. Male rats were primed with estradiol and functional dopaminergic prolactin release-inhibiting factor receptors were completely blocked by pretreatment with a large dose of pimozide (3 mg/kg), a dopaminergic receptor blocking agent. The neurophysin-II stimulated prolactin release in the rats which did not have any functional dopaminergic prolactin release-inhibiting factor receptors suggesting that neurophysin-II likely initiates a chain of events which eventually stimulates prolactin-releasing factor release since the possibility of involvement of the dopaminergic prolactin release-inhibiting factor system is eliminated. Opioids are known to be one of a chain of events which transmit external stress into a stimulation of prolactin release. Naloxone, a μ-receptor antagonist, was injected 20 min before neurophysin-II administration into rats which were primed with estradiol and pretreated with pimozide (3 mg/kg), but the naloxone administration did not block the prolactin release stimulated by neurophysin-II injection. This result indicates that opioids are not one of the chain of events between initiation of stimulation by neurophysin-II and prolactin release.
Parathyroid hormone potentiates the effect of insulin-like growth factor-I on bone formation
Authors: Spencer E. Martin , Si Erwin C. C. , Liu Chung C. , Howard Guy A. 
Published: December 2008

Abstract.
Insulin-like growth factor-I and parathyroid hormone are both known regulators of bone formation. In this study, human recombinant IGF-I and bovine PTH (1–34) and their combination were studied for their effects in vitro on the proliferation of embryonic chick osteoblast-like cells (osteoblasts) and in vivo on bone formation in normal rats. Osteoblasts from 17-day-old chick embryos were cultured in serum-free BGJb medium containing 0.1% bovine albumin. After 2 days, IGF-I and/or PTH were added. Twenty-four hours later [3H]thymidine incorporation into trichloroacetic acid precipitable material was quantified as an index of cell proliferation. This has previously been shown to reflect actual cell division. IGF-I at doses ranging from 0.85 to 13.6 nmol/l caused a dose-dependent increase in [3H]thymidine incorporation into osteoblasts. PTH alone (10 to 1000 pmol/l) had no significant effect. However, when combined with IGF-I, PTH potentiated the mitogenic effect of IGF-I and achieved statistical significance at 30 and 100 pmol/l (p <0.05). This potentiation was also studied in vivo. The right hindlimbs of rats weighing 150 g were infused intra-arterially by an osmotic minipump with graded doses of IGF-I (0.1 to 0.4 nmol/day) and/or PTH (0.27 nmol/day) for 7 days. The rate of trabecular bone apposition (formation) was measured by double tetracycline labelling and compared with the contralateral uninfused limb which acted as the control. Histomorphometric data revaled that neither IGF-I nor PTH alone had a significant effect on trabecular bone apposition rate compared with control limbs. The co-infusion of IGF-I (0.4 nmol/day) and PTH (0.27 nmol/day) resulted in a marked increase in trabecular bone apposition rate. The results of 2 studies were significant at p < 0.01. These data suggest that PTH potentiates the effect of IGF-I on bone formation both in vivo and in vitro.
The role of thyroid hormone autoantibodies in serum transport
Authors: Copping Susan , Byfield Peter G. H. 
Published: December 2008

Abstract.
Eleven sera known to contain thyroid hormone autoantibodies were analysed by reverse-flow electrophoresis for the equilibrium distribution of thyroid hormones between these autoantibodies and the three normal binding proteins found in serum. The binding properties of the autoantibodies determined in vitro did not necessarily predict their contribution to transport in serum of T4 and T3. Some could both bind in vitro and transport in serum. Others were able to bind both hormones but transported only one. However, some autoantibodies could be specific, binding and transporting one hormone only. In some sera, the autoantibody was the dominant transport protein having drawn hormone from thyroxine-binding globulin which is normally the most important. The autoantibodies were not saturated even in euthyroid individuals, indicating that they bind hormone reversibly and are a part of an equilibrium system.
Extragonadal effects of luteinizing hormone in mice
Authors: Nomura Kaoru , Puett David W. , Puett David , Shizume Kazuo , Liddle Grant W. 
Published: December 2008

Abstract.
LH is composed of isoforms which exhibit microheterogeneity. We recently demonstrated that a particular ovine or porcine LH preparation (G100-fr.3) stimulates kidney growth. This study was conducted to clarify the physiological role of this renotropic activity and other extragonadal effects of the ovine LH preparation in CD-1 mice. Hypophysectomy caused a significantly greater reduction in relative dry kidney weight (i.e. g/100 g body weight) when compared to adrenalectomy, castration, thyroidectomy, and castration plus thyroidectomy. Supplementation with G100-fr.3 in these animals partially restored not only kidney size but also DNA, RNA and protein content. Treatment with standard LH preparations (NIDDKoLH24 and G3-268DA), as well as PRL, GH, FSH and TSH, failed to reverse the renal atrophy induced by hypophysectomy and castration. Administration of testosterone to castrated hypophysectomized mice increased kidney weight and RNA content, but not renal DNA. The relative dry kidney weight increased significantly at the onset of puberty in intact male mice, but not in castrated males or intact female mice. In addition, human CG increased kidney size in hypophysectomized male mice, but not in castrated hypophysectomized animals. These findings indicate that LH isoforms may regulate kidney growth in the male mouse both directly as a renotropin stimulating hyperplasia and indirectly as a gonadotropin via testicular androgen, producing cellular hypertrophy. It was also noted that G100-fr.3 decreased hepatic weight, DNA, RNA and protein, but produced no significant change in the spleen, heart or adrenal glands in castrated-hypophysectomized mice. Such extragonadal effects of G100-fr.3 were also observed in intact female mice. These results suggest that certain LH isoforms may have extragonadal actions involving the kidney and liver.
Adrenal function in HIV infected patients
Authors: Verges B. , Chavanet P. , Desgres J. , Vaillant G. , Waldner A. , Brun J. M. , Putelat R. 
Published: December 2008

Abstract.
Since anatomopathological lesions of the adrenal gland have been frequently observed at autopsy in AIDS, we investigated the glucocorticoid function in 63 patients (51 men, 12 women) infected by the human immunodeficiency virus (HIV) in order to determine the incidence and the nature of any adrenocortical abnormalities at various stages of HIV infection. The patients were classified according to the Centers for Disease Control (CDC) recommendations into group II (asymptomatic; N = 13), group III (lymphadenopathy; N = 27) and group IV (clinical manifestations; N = 23). Plasma ACTH and cortisol before and after an exogenous ACTH stimulation test were measured in patients as in 30 agematched controls. Plasma renin activity and plasma aldosterone before and after ACTH stimulation were also measured in 31 patients (group II: 12; group III: 10; group IV: 9). Compared with controls patients from group II–III had higher levels of ACTH (39.11 ± 17.01 vs 29.73 ± 8.53 ng/l; p = 0.003) and basal cortisol (232 ± 91.2 vs 184.3 ± 30.9 μg/l; p = 0.03). No significant differences were noted between group IV patients and controls as to ACTH and basal and stimulated cortisol levels. Among the 63 patients, only one from group IV had a blunted cortisol response after ACTH stimulation test. Plasma renin activity, and basal and stimulated aldosterone levels in the 3 groups of patients were not different from control values. In conclusion: 1. Adrenal insufficiency does not seem very frequent in group IV patients and is likely to be a late complication in AIDS. 2. The increased ACTH and basal cortisol levels found in group II and group III patients argue for an early dysregulation of the adrenocortical axis in HIV infection. The exact physiopathological mechanism is not yet known, but an enhanced CRH production by interleukin 1 and/or a direct role of the HIV envelope glycoprotein (gp 120) may explain the high ACTH level in HIV patients.
Low basal thyrotropin with normal thyroid function in primary hyperparathyroidism
Authors: Gillet Clementine , Bergmann Pierre , Francois Dominique , Body Jean-Jacques , Corvilain Jacques 
Published: December 2008

Abstract.
TSH serum levels and thyroid function in 32 patients with primary hyperparathyroidism and hypercalcemia were compared to those of 30 age and sex-matched normal subjects. Serum T3 and T4 concentrations in hyperparathyroidism were not different from normal. However, basal serum TSH concentrations measured with an ultrasensitive immunoradiometric assay were significantly lower than normal (1.09 ± 0.49 vs 2.06 ± 0.85 mU/l, p < 0.001). In hyperparathyroidism, TSH, but not T4 or T3, was negatively correlated with serum calcium, not with iPTH. The increase in TSH (Δ TSH) 30 min after the iv injection of TRH was also significantly blunted in patients with primary hyperparathyroidism; Δ TSH was highly correlated with basal TSH in hypercalcemic patients. The basal TSH concentration was higher and no longer different from normal (1.70 ± 1.2 mU/l) 2 to 12 months after removal of the parathyroid adenoma, when serum calcium was normalized, whereas T3 and T4 did not change. A low basal TSH with normal T4 and low T3 was found in 13 patients with hypercalcemia of malignancy. In these patients, TSH increased after treatment of hypercalcemia with 3-amino-1,hydroxypropylidene-1,1-bisphosphonate, whereas T4 did not change. The results suggest that the set point of pituitary thyroid feedback control could be decreased in chronic hypercalcemia and that hypercalcemia could render the thyroid more sensitive to TSH.
Activity of insulin receptor kinase and glycogen synthase in skeletal muscle from patients with chronic renal failure
Authors: Bak Jens F. , Schmitz Ole , Sørensen Søren S. , Frøkjær Jens , Kjær Torben , Pedersen Oluf 
Published: December 2008

Abstract.
To examine subcellular mechanisms behind the pathogenesis of peripheral insulin resistance in chronic uremic patients, insulin receptor function and glycogen synthase activity were studied in biopsies of skeletal muscle obtained during renal transplant surgery in 9 non-diabetic uremic patients. The results were compared with values obtained in an age- and sex-matched group of subjects with normal renal function, undergoing surgery for urological or gynecological diseases. The recovery of solubilized, wheat germ agglutinin-purified insulin receptors from skeletal muscle was increased among the uremic patients: 49.3 ± 6.1 vs 31.4 ± 2.8 fmol/100 mg muscle in healthy controls (p < 0.03). Basal as well as insulin-stimulated kinase activities of the insulin receptors, expressed as phosphorylation of the synthetic peptide poly(Glu-Tyr(4:1)) were similar. In addition, the maximal activity of the glycogen synthase was enhanced in uremic muscle: 26.6 ± 2.8 vs 19.5 ± 1.8 nmol · (mg protein)−1 · min−1 (p < 0.05), whereas the half-maximal activation constant for glucose-6-phosphate was identical in the two groups. Likewise, the muscle glycogen concentrations were similar in the uremic patients and the normal controls. In conclusion, our data suggest that neither impaired insulin receptor function nor a reduced maximal glycogen synthase activity of skeletal muscle are involved in the pathogenesis of the insulin resistance of patients with chronic renal failure.
Long-term effects of radiotherapy for acromegaly on circulating prolactin
Authors: Ciccarelli E. , Corsello S. M. , Plowman P. N. , Jones A. E. , Touzel R. , Rees L. H. , Besser G. M. , Wass J. A. H. 
Published: December 2008

Abstract.
In 61 acromegalic patients, serum PRL was assessed (off medical treatment) before and 2 to 12 (mean 6.4) years after external beam radiotherapy. Before radiotherapy elevated PRL levels were present in 22 of 35 males (63%) and 12 of 26 females (46%) and were above 1000 mU/l in 11 males and 5 females. When studied for up to 5 years after radiotherapy, 22 of 23 (96%) patients who had not had surgery and who had normal PRL pre-radiotherapy showed an increased PRL level and this was also seen in 17 of 27 (63%) who had been hyperprolactinaemic initially. In contrast, 10 of 27 patients (37%) who had elevated pre-radiotherapy levels (all greater than 1000 mU/l) had a reduction in PRL values after radiotherapy. In all 11 patients who underwent surgery before radiotherapy, an increase in PRL was seen after radiotherapy. In the 21 patients followed for 10–12 years, the peak PRL value occurred 1–16 years after radiotherapy. After this, a progressive reduction of PRL to normal was seen. Normal levels were reached 4 to 10 years after radiotherapy. No correlation was found between pre-treatment PRL values and final GH values in the whole group, nor between changes in PRL and the development of impaired ACTH or TSH secretion. Thus, different patterns of PRL behaviour suggest that radiotherapy treatment may either produce hyperprolactinemia from mild hypothalamic damage or ablate PRL secreting cells if they were present in the tumour before treatment. These changes do not predict final GH results or the development of hypopituitarism after radiotherapy.
Persistent impairment of the insulin response to glucose both in vivo and in vitro after streptozotocin exposure: Studies with grafted pancreatic islets and islets maintained in culture
Authors: Eizirik Décio L. , Sandler Stellan , Korsgren Olle , Jansson Leif , Andersson Arne 
Published: December 2008

Abstract.
The functional responses of the pancreatic B-cells after cytotoxic damage are still largely unknown. Using in vitro models to clarify this issue, we have recently observed a preferential reduction of glucose-stimulated insulin production and release in mouse pancreatic islets maintained in culture after in vitro exposure to streptozotocin. In order to evaluate the relevance of these findings in vivo, two sets of experiments were performed. First, mouse pancreatic islets were exposed in vitro to 2.2 mmol/l streptozotocin or vehicle alone, cultured for 6 days, and finally grafted under the kidney capsule of normoglycemic nude mice. Two weeks after transplantation there was no difference in the total DNA and insulin content between the two groups of islet grafts, but the insulin concentration, as expressed per μg DNA, was decreased by 40% in the streptozotocin-treated islets. The insulin release of the grafts, during perfusion of the graft-bearing kidney in situ with 16.7 mmol/l glucose was diminished in the streptozotocin group, whilst perfusion with 16.7 mmol/l glucose plus 5 mmol/l theophylline was able partially to counteract the reduction in insulin release. In the second set of experiments, NMRI mice were injected iv with 160 mg/kg streptozotocin or vehicle alone, and their islets isolated 15 min after the injections. After 6 days in culture, there was no decrease in DNA, glucagon and somatostatin contents, but the insulin content was decreased by 40% in the streptozotocin exposed islets. These islets also showed a 60% decrease in the insulin response to glucose, which was partly counteracted by incubation with 16.7 mmol/1 glucose plus 5 mmol/l theophylline. These observations suggest that a defective response to glucose, in conjunction with a better response to non-nutrient secretagogues, may be a common property of pancreatic islets following toxin-induced disturbances.