bluuebunny/crossref_metadata_2025_split huggingface dataset:
Antiuterotrophic effects of the pure antioestrogen ICI 182,780 in adult female monkeys (Macaca nemestrina): quantitative magnetic resonance imaging
Authors: Dukes M. , Waterton J. C. , Wakeling A. E. 
Published: December 2008

ABSTRACT
The antiuterotrophic efficacy of the pure antioestrogen ICI 182,780 has been demonstrated previously by magnetic resonance imaging (MRI) in ovariectomized oestrogen-treated monkeys (Macaca nemestrina). Further characterization of the effects of ICI 182,780 in intact adult female monkeys with normal menstrual cycles was undertaken to provide an indication of its potential actions in premenopausal women. Changes in the volume of uterine tissues were measured by MRI in early, mid and late cycle. The volume of the uterus varied up to fivefold between individual monkeys but serial observations in individuals provided sufficient precision to allow accurate assessments to be made of changes in the endometrium and myometrium during the course of the menstrual cycle and following ICI 182,780 administration. In comparison with its initial size in untreated monkeys, the endometrium increased in volume by 60% and 125% in the mid and late cycle respectively. In contrast, the size of the myometrium decreased significantly, by 16% from early to mid cycle and then recovered to near its initial volume in the late cycle. Treatment with ICI 182,780 beginning in the early part of the menstrual cycle prevented the growth of the uterus. The magnitude and duration of the response was dependent on whether or not ovulation occurred during treatment with ICI 182,780. In animals rendered anovulatory, growth of the endometrium was blocked completely by ICI 182,780 and the volume of the tissue declined below that present at the start of the menstrual cycle. Antiuterotrophic efficacy was significantly less in monkeys which ovulated during treatment with ICI 182,780. The volume of myometrium was reduced substantially by antioestrogen treatment, and the difference in response between ovulatory and anovulatory monkeys was less marked than that of the endometrium.
Journal of Endocrinology (1993) 138, 203–209
Distinct regional localization of neurocalcin, a Ca2+-binding protein, in the bovine adrenal gland
Authors: Nakano A. , Terasawa M. , Watanabe M. , Okazaki K. , Inoue S. , Kato M. , Nimura Y. , Usuda N. , Morita T. , Hidaka H. 
Published: December 2008

ABSTRACT
Neurocalcin (molecular weight 23 000 and 24 000) is a Ca2+-binding protein with three putative Ca2+-binding domains and is present in large amounts in nervous tissues. Neurocalcin isoproteins separated by C18 reverse-phase column chromatography are insoluble in buffer solution and it is impossible to determine the dissociation constant of neurocalcin with Ca2+. To overcome this difficulty, recombinant neurocalcin was synthesized, based on one of the cDNAs of the neurocalcin isoproteins. Stoichiometric titration experiments, using recombinant neurocalcin, indicated that this protein bound 2 mol Ca2+/mol protein and that the apparent dissociation constant for Ca2+ was 2·2 μmol/l, suggesting that neurocalcin plays a physiological role in cellular function. Immunoblotting showed that neurocalcin is present in the bovine adrenal gland in addition to the nervous tissues. Neurocalcin, identified by immunoblotting, was purified from the bovine adrenal gland. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of neurocalcin from the bovine brain showed 23 kDa and 24 kDa double bands, while SDS-PAGE of neurocalcin from the adrenal gland showed a single band of apparently 24 kDa, suggesting that the expression of neurocalcin isoproteins differs from tissue to tissue. The content of neurocalcin in the adrenal gland was 10 μg protein/100 g wet tissue. Immunohistochemical analysis showed the occurrence of neurocalcin in zona glomerulosa and adrenal medulla but not in zona fasciculata or zona reticularis. The restricted localization of neurocalcin in the adrenal gland suggests that a similar Ca2+ signal pathway may be present in zona glomerulosa and the adrenal medulla.
Journal of Endocrinology (1993) 138, 283–290
Renal melatonin excretion in sheep is enhanced by water diuresis
Authors: Valtonen M. , Laitinen J. T. , Eriksson L. 
Published: December 2008

ABSTRACT
Diurnal variation in blood melatonin levels and renal melatonin excretion was monitored in five ewes by blood sampling and quantitative urine collection at 2-h intervals. A typical secretory pattern of melatonin was seen both in blood and urine levels and in the renal excretion of melatonin. Serum melatonin levels increased from daytime values of approximately 200 pmol/l to a mean of 800 pmol/l during darkness. Urine flow rate and urine osmolality did not show any clear diurnal rhythm.
To examine whether urine flow rate affects renal melatonin excretion at night, urine was collected in three consecutive 30-min fractions, and blood was sampled in the middle of each urine collection period when the sheep were in normal water balance or after hydration. Hydration increased urine flow rate over sixfold and decreased urine osmolality well below plasma osmolality. Glomerular filtration rate, measured as creatinine clearance, did not change. Serum melatonin concentrations did not differ between hydrated and non-hydrated sheep. However, urinary melatonin excretion was 1·1 ± 0·3 (s.e.m.) pmol/min at midnight in normal water balance, and significantly higher (2·6 ± 0·4 pmol/min) in the hydrated state.
In this study, the validity of urinary melatonin determinations as an indicator of pineal function was confirmed in normal water balance. In addition, our results suggest that a high tubular fluid load during diuresis increases urinary melatonin excretion because of decreased tubular reabsorption. This means that urine flow rate should be taken into consideration in studies where urinary melatonin levels are used as an index of production of the indole by the pineal gland.
Journal of Endocrinology (1993) 138, 445–450
Proteolytic modification of insulin-like growth factor-binding proteins: comparison of conditioned media from human cell lines, circulating proteases and characterized enzymes
Authors: Frost V. J. , Macaulay V. M. , Wass J. A. H. , Holly J. M. P. 
Published: December 2008

ABSTRACT
Proteolytic modification of circulating insulin-like growth factor binding protein-3 (IGFBP-3) has been described in a number of conditions. Using Western ligand blotting and SDS-PAGE analysis of fragmentation patterns of 125I-labelled IGFBP-3 and 125-labelled IGFBP-1, we have examined conditioned media from cultured human cell lines for the presence of proteolytic activity and compared this with the action of circulating proteases and with characterized enzymes including cathepsin D, kallikrein, plasmin and tissue plasminogen activator.
125I-labelled IGFBP-3 was incubated with serum from pregnant women, patients following heart surgery and patients with cancer of the breast, lung or head/neck. Following separation of the preincubated samples by SDS-PAGE, a distinct pattern of degradation fragments was observed which was similar in all cases. This proteolytic activity was inhibited in the presence of EDTA, phenanthroline and 4(-2-aminoethyl)-benzenesulphonylfluoride,HCl. These proteases had no detectable effect on IGFBP-1. Serum-free conditioned medium from a human dermal fibroblast cell line, a rabdomyosarcoma, a cervical, a bladder, a chorio- and two-tongue squamous cell carcinoma cell lines all contained proteolytic activity which fragmented IGFBP-3. The pattern of fragments was similar in all cell lines but different from that produced by the circulating proteases. Six out of nine cell lines produced protease(s) which degraded IGFBP-1 in addition to IGFBP-3. Whilst all the characterized enzymes tested also fragmented IGFBP-3 and plasmin cleaved IGFBP-1, none of these acted in the same way as either circulating or cell line-derived proteolytic activity. The activity associated with the characterized enzymes and cell lines was inhibited in the presence of serum from normal healthy subjects.
These results demonstrate that the serum of pregnant women, post-operative patients and patients with cancer contain circulating proteases which cause fragmentation of IGFBP-3 but have little effect on IGFBP-1. Cell-derived proteases were shown to act on IGFBP-3 and IGFBP-1 in a number of instances but are not active in the presence of circulating inhibitors. These proteases may play an important role in regulating the availability of IGFs to normal and neoplastic tissues.
Journal of Endocrinology (1993) 138, 545–554
Increased cytosolic free sodium concentrations in platelets from type 2 (non-insulin-dependent) diabetic patients is associated with hypertension
Authors: Tepel M. , Bauer S. , Husseini S. , Raffelsiefer A. , Zidek W. 
Published: December 2008

ABSTRACT
Cytosolic free sodium concentrations ([Na+]i) in intact platelets from 32 type 2 (non-insulin-dependent) diabetic patients and from 27 age- and sex-matched non-diabetic control subjects were measured with the novel sodium-sensitive fluorescent dye sodium-binding-benzofuran-isophthalate. [Na+]i was significantly higher in platelets from type 2 diabetic patients compared with control subjects (40·6 ± 2·4 vs 32·0 ± 2·0 mmol/l, means ± s.e.m., P<0·03). Both systolic and diastolic blood pressure were significantly elevated in diabetic patients compared with control subjects. Analysis of diabetic patients showed a significant association between [Na+]i and diastolic blood pressure (P =0·026). Stimulation of Na/H exchange by thrombin increased [Na+]i in both groups. After inhibition of Na/K/ATPase by ouabain (1 mmol/l), [Na+]i was significantly increased both in diabetic patients and non-diabetic subjects in a similar way (by 40·2 ± 7·3 and 31·7 ± 5·3 mmol/l respectively). It is concluded that increased [Na+]i in cells from type 2 diabetic patients may be related to hypertension.
Journal of Endocrinology (1993) 138, 565–572
Thyroidectomy progressively renders the reproductive system of starlings (Sturnus vulgaris) unresponsive to changes in daylength
Authors: Dawson A. 
Published: December 2008

ABSTRACT
If starlings (Sturnus vulgaris) are transferred to long days soon after being thyroidectomized, they show gonadal maturation but no subsequent photorefractoriness, suggesting a specific role for thyroid hormones in the development of photorefractoriness. This study tests an alternative hypothesis: that following thyroidectomy all gonadal responses to photoperiodic changes are progressively inhibited. Photoperiodic responses of starlings were examined at two different times after thyroidectomy. Transfer of thyroidectomized birds to long days 4 weeks after thyroidectomy caused testicular growth but at a slower rate than in intact birds, whereas, in thyroidectomized birds kept on short days, testicular growth was greater than in corresponding intact birds. Thirteen weeks after thyroidectomy, there was little or no response to an increase or a decrease in daylength in thyroidectomized birds, whereas intact birds showed marked testicular growth or regression respectively. The results support the hypothesis that the reproductive system gradually becomes unresponsive to changes in daylength during the weeks following thyroidectomy. This long-term effect may mean that earlier conclusions regarding the effects of thyroidectomy need to be reassessed.
Journal of Endocrinology (1993) 139, 51–55
Biochemical and immunological characterization of oestrogen receptor in the cytosolic fraction of gluteal, omental and perirenal adipose tissues from sheep
Authors: Watson G. H. , Manes J. L. , Mayes J. S. , McCann J. P. 
Published: December 2008

ABSTRACT
Determination of the presence and characterization of oestrogen receptors (ERs) in subcutaneous and internal fat depots were performed and compared with ERs in the uterus using ligand binding and immunological techniques. Successful and consistent measurement of ERs in ovine adipose tissue could only be accomplished in animals depleted of endogenous sex steroids by combined ovariectomy and adrenalectomy. Scatchard, sucrose gradient and Western blot analyses all confirmed the presence of ERs in the cytosolic fractions of various adipose and uterine tissues from ovariectomized-adrenalectomized ewes. The approximate Kd values of 0·1–0·4 nmol/l for oestradiol binding in cytosolic fractions of gluteal, omental and perirenal adipose tissues were similar to the expected high affinity binding of Kd 0·35 nmol/l observed in uterine tissue. The binding was specific for oestrogens, as unlabelled diethylstilboestrol and oestradiol effectively competed with labelled hormone for receptor sites and progesterone, R5020, testosterone and dexamethasone all failed to compete. Mean (± s.e.m.) concentrations of ERs, expressed as fmol specific binding sites per mg protein, were much lower (P<0·05) in adipose tissues than in uterine tissue (975 ± 33). However, the content of ERs was greater (P<0·05) in subcutaneous gluteal fat (11·5 ± 0·8) than in the internal omental or perirenal fat (5 ± 0·6) depots. ERs from adipose and uterine tissues both migrated as moieties of 8S on 5–20% sucrose gradients. Western blot analysis of ERs from uterine and adipose tissues in the presence of protease inhibitors demonstrated an immunostaining band with a molecular mass of 67 kDa. In the absence of protease inhibitors, the adipose ER was degraded into two smaller bands with molecular masses of 45 kDa and 36 kDa.
In conclusion, these results support the presence of specific ERs in adipose tissues of sheep with characteristics similar to those of the ERs of the uterus. Furthermore, their presence is supportive of sex steroid-dependent effects on adipose accretion and metabolism.
Journal of Endocrinology (1993) 139, 107–115
Preferential increase in prostaglandin endoperoxide H synthase compared with lipoxygenase activity in sheep placenta and amnion at term pregnancy and after intrafetal glucocorticoid administration
Authors: Langlois D. A. , Fraher L. J. , Khalil M. W. , Fraser M. , Challis J. R. G. 
Published: December 2008

ABSTRACT
Prostaglandins (PGs) have been implicated as stimulants to myometrial contractility at parturition in many species. To determine whether the increased production of PGs at parturition reflects a general increase in the metabolism of arachidonic acid or a specific increase in PG endoperoxide H synthase (PGHS) compared with lipoxygenase activities, and to determine intrauterine sites of these activities, we examined the metabolism of [3H]arachidonic acid by homogenates of placenta, amnion and chorion from sheep at days 78–80, 100–105, 135–140 of pregnancy and at term (day 145). Tissues were also obtained from fetuses at day 125; four of these were infused for 84 h with cortisol and four were used as saline-treated controls. The endogenous arachidonic acid content at the start of incubation was measured by capillary gas chromatography. Radioactive metabolites were separated and quantified by reverse-phase high-pressure liquid chromatography.
At each gestational age arachidonic acid was converted to PGs, leukotrienes (LTs) and hydroxyeicosatetraenoic acids (HETEs). Conversion to PGs was greater in amnion than in chorion or placenta between days 78 and 140. The formation of PGs rose in placenta at term to a mean value twice that of amnion and ten times that of chorion. In amnion, the ratio of PG: LT rose significantly at term relative to 100–140 days of gestation. In placenta, the ratio of PG: LT produced from arachidonic acid and the ratio of total PGHS: lipoxygenase products rose significantly at term. In the day-125 fetuses treated with cortisol there was a significant increase in PG production relative to that in control fetuses infused with saline in placenta, amnion and chorion; the placenta and amnion being the major sites of PG production. Production of LTs and HETEs also rose significantly in the chorion and the placenta relative to controls. In both the placenta and the amnion there was a significant increase in the ratio of total PGHS to lipoxygenase products formed. We conclude that at term labour and in labour induced by intrafetal cortisol infusion, the placenta is the major site of arachidonic acid metabolism, and that there is a preferential increase in the formation of PGs over lipoxygenase products. These results are consistent with the suggestion that there is an increase in the expression or activity of PGHS in the placenta of sheep in late pregnancy.
Journal of Endocrinology (1993) 139, 195–204
Suppression of basal secretion of FSH inhibits follicular development and maturation during the oestrous cycle of the rat
Authors: Noguchi J. , Watanabe G. , Taya K. , Sasamoto S. 
Published: December 2008

ABSTRACT
The role of basal FSH secretion during the rat oestrous cycle in regulating ovulation was examined by suppressing FSH secretion using charcoal-treated porcine follicular fluid (pFF). Although 0·5 ml pFF given at 05.00 and 11.00 h on the day of pro-oestrus had no effect on ovulation, 0·5 ml pFF given five times at 6-h intervals from 11.00 h on dioestrus to 11.00 h on pro-oestrus completely eliminated ovulation on the morning of the next oestrus. When 0·25 ml pFF was given on the same schedule, all animals ovulated a significantly decreased number of oocytes (9·0 ± 0·8) at the next oestrus. During the period of pFF treatment, the number of follicles capable of ovulating in response to human chorionic gonadotrophin (hCG) decreased (7·6 ± 0·7 at 05.00 h on pro-oestrus) and plasma levels of oestradiol showed a peak level 6 h later than in controls treated with 0·5 ml steroid-free porcine serum. Treatment with pFF suppressed plasma FSH concentrations in a dose-dependent manner, although plasma LH was inhibited irregularly. Supplementary administration of LH throughout the treatment period with 0·5 ml pFF resulted in ovulation of one to three oocytes in response to hCG in only three out of ten animals. These results suggest that basal secretion of FSH during the rat oestrous cycle plays an essential role in follicular development and maturation towards ovulation, and that the levels of FSH secretion may contribute to the maturation of normal numbers of follicles for ovulation.
Journal of Endocrinology (1993) 139, 287–293
Monoclonal antibodies to bovine growth hormone potentiate hormonal activity in vivo by enhancing growth hormone binding to hepatic somatogenic receptors
Authors: Massart S. , Maiter D. , Portetelle D. , Adam E. , Renaville R. , Ketelslegers J.-M. 
Published: December 2008

ABSTRACT
Administration of GH complexed with monoclonal antibodies (MABs) potentiates the in vivo actions of the hormone. In particular, growth and serum IGF-I concentrations of GH-treated hypophysectomized rats are increased by concomittant injection of anti-GH MABs. Among 37 anti-bovine GH (bGH) MABs, we selected one MAB with the most potentiating effects to investigate the mechanisms responsible for this phenomenon. Hypophysectomized rats were killed 18 h after a single s.c. injection of bGH (100 μg/rat), alone or complexed with increasing doses of MAB (4, 40, 400 μg/rat; MAB:bGH molar ratio: 0·005, 0·05, 0·5). IGF-I was measured by radioimmunoassay in acid–extracted sera and livers, whereas liver IGF-I mRNA was quantified by Northern blot hybridization. The in vivo occupancy of liver somatogenic (GH) receptors was derived from the determinations of total and free 125I-labelled bGH binding to liver homogenates treated with 4 mol MgCl2/l or water. Injection of MAB–bGH complexes enhanced body weight gain and raised serum IGF-I, liver IGF-I and liver IGF-I mRNA more than bGH alone (1·6-, 6-, 10- and 7-fold increases at the highest dose of MAB, compared with bGH alone; P < 0·001). These potentiating effects of the MAB were dose-dependent and significant potentiation of the growth response was already observed with the lowest dose of MAB. In vivo occupancy of liver GH receptors was markedly higher 18 h after injection of MAB–bGH complexes than after bGH alone, and this effect was also dose-dependent (receptor occupancy of 28%, 37% and 83% after 4, 40 and 400 μg of MAB respectively compared with 6% after bGH alone; P < 0·05, 0·05 and 0·001 respectively). In contrast, the in vitro binding of 125I-labelled bGH to liver homogenates was decreased in the presence of high doses of MAB.
We conclude that low amounts of MABs complexed with bGH potentiate the stimulation by the hormone of liver IGF-I synthesis and secretion in a dose–dependent manner. These effects are mediated, at least in part, through changes in hormone-receptor interaction in vivo, leading to enhanced and/or prolonged binding of bGH to its somatogenic receptors.
Journal of Endocrinology (1993) 139, 383–393
Effect of a β3-adrenergic agonist, BRL35135A, on glucose uptake in rat skeletal muscle in vivo and in vitro
Authors: Abe H. , Minokoshi Y. , Shimazu T. 
Published: December 2008

ABSTRACT
The effects of the β3-agonist, BRL35135A, on glucose uptake in the peripheral tissues of the rat, including skeletal muscle, were studied using the 2-[3H]deoxyglucose method in anaesthetized adult animals. Intravenous infusion of the β3-agonist dose-dependently increased the rate constant of glucose uptake in three types of skeletal muscle, brown adipose tissue, white adipose tissue, heart and diaphragm, but not in the brain, spleen or lung. Although infusion of the β3-agonist did not change the plasma concentration of glucose appreciably, it caused an increase in the plasma concentration of insulin when given at more than 25 μg/kg per h. To ascertain whether the effect of the β3-agonist on glucose uptake in skeletal muscle is mediated by insulin, glucose uptake into soleus muscle isolated from young rats was also measured in vitro using different concentrations of the β3-agonist. The β3-agonist BRL37344 (an active metabolite of BRL35135A) significantly increased glucose transport in a dose-dependent manner, with maximum stimulation at 100 pmol/l. These results demonstrate that glucose uptake in skeletal muscle can be enhanced independently of the action of insulin, probably through the mediation of β3-adrenoceptors present in the tissue.
Journal of Endocrinology (1993) 139, 479–486
Interaction of monoclonal antibodies with growth hormone-binding protein and its complex with growth hormone
Authors: Sadeghi H. , Lumanglas A. L. , Baumbach W. R. , Wang B. S. 
Published: December 2008

ABSTRACT
The properties of four independent lines of monoclonal antibodies (MAbs) specific to rat GH-binding protein (GHBP) were examined. Three MAbs, designated GHR-12, GHR-13 and GHR-16, were raised against the entire GHBP molecule. The fourth MAb, designated as GHBP4·3, was raised against the 17 amino acid residues at the C-terminal end of rat GHBP. The interaction of these antibodies with GHBP and their effect on GH binding to GHBP were analysed by conventional competition binding assays and surface plasmon resonance, i.e. with a Biospecific Interaction Analysis (BIAcore) instrument. The binding affinity of these MAbs to GHBP ranged from 29 nmol/l to 30·9 pmol/l. The pair-wise antibody binding to GHBP on BIAcore suggested that GHR-13 and GHR-16 recognized different antigenic determinants while part of the GHR-12 epitope might be shared with the other antibodies. The antibodies inhibited the interaction of GH with GHBP in the competition binding assay. However, in sequential binding on the BIAcore instrument, they were able to bind GHBP after its interaction with GH, indicating that the inhibition observed in the competition binding assay resulted from steric hindrance rather than direct interference with the GH-binding site of GHBP. The present findings, therefore, suggest that these antibodies are useful for investigating GHBP and its interaction with GH.
Journal of Endocrinology (1993) 139, 495–501