bluuebunny/crossref_metadata_2025_split huggingface dataset:
Molecular heterogeneity of the β-core fragment of human chorionic gonadotrophin
Authors: de Medeiros S. F. , Amato F. , Matthews C. D. , Norman R. J. 
Published: December 2008

ABSTRACT
We have analysed the structure and composition of the β-core fragment of human chorionic gonadotrophin (βC-hCG) from fresh urine specimens obtained from pregnant women and compared our findings with those previously proposed by other groups using different protocols. SDS-PAGE separation of reduced βC-hCG demonstrated two major bands with apparent molecular weights of Mr 8900 and Mr 7500. The molecular weight of the agalacto βC-hCG was estimated to be Mr 10 218 from the amino acid analysis after high-performance liquid chromatography (HPLC) separation. Moreover, HPLC separation of its reduced and S-carboxymethylated peptides resulted in three peaks, but only two of them could be sequenced and demonstrated to be the previously reported β6-40 (Mr 5000) and β55-92 (Mr 5300) peptides of the βhCG subunit. The results showed that 56-78% of βC-hCG molecules of molecular weight Mr 12 800 were able to bind Concanavalin A (Con A). While most were lacking all the peripheral monosaccharides and terminated in mannose, some retained other sugar residues on their antennae. Direct carbohydrate analysis showed the following molar content normalized to six mannose molecules: galactose 2·8, glucosamine 5·3, galactosamine 0·3, fucose 1·7 and sialic acid 3·0. Approximately 22–44% of the βC-hCG molecules did not bind Con A (Con A non-reactive forms), of which 88% were totally deprived of sugar units and had an apparent molecular weight of approximately Mr 10 000, and 12% were weakly reactive to Con A and reactive to anion exchange (negatively charged forms), being incompletely trimmed of their oligosaccharide chains. Comparison of our results with those of two other groups have indicated that the differences noted among preparations are due to either the source or the methods used to purify and characterize this fragment. In addition, our results showed significant microheterogeneity on the N-linked oligosaccharide moieties with some molecules apparently having no sugar molecules. These results have implications for the origins of βC-hCG, suggesting secretion of some molecules without sugar chains and in other cases possible metabolism of hCG in the peripheral tissues.
Journal of Endocrinology (1993) 139, 519–532
Changes in hepatic insulin-like growth factor-binding proteins -1, -2 and -3 mRNA levels in rats with altered thyroid status
Authors: Rodriguez-Arnao J , Miell J , Thomas M , McGregor A M , Ross R J M 
Published: December 2008

Abstract
Changes in thyroid status have a major effect on the GH/insulin-like growth factor (IGF) axis. The majority of IGF in the circulation is bound to specific IGF-binding proteins (IGFBPs) of which six have been cloned and sequenced. We have studied changes in hepatic gene expression of IGFBP-1, -2 and -3, in male Wistar rats rendered hyperthyroid (thyroxine, 200 μg/kg per day) or hypothyroid (propylthiouracil, 0·1% daily). Littermates of the same age were used as controls (n=6 in each group). Thyroxine was measured by radioimmunoassay, and hepatic IGFBP-1, -2 and -3 mRNA levels by Northern blot analysis using specific rat cDNA probes with a 28S ribosomal probe as a loading control. Mean± s.e.m. thyroxine levels were 247·0±44·5 (hyperthyroid group), <9·0 (hypothyroid group) and 76·0 ± 4·5 nmol/l (control group). IGFBP-1 and -2 mRNA levels in the hypothyroid animals compared with the controls were significantly increased, but similar levels of expression were found in thyrotoxic and control rats. IGFBP-3 mRNA levels in hypothyroid animals were decreased, and increased in thyrotoxic animals. Thus, in the adult rat, hypothyroidism is associated with increased hepatic IGFBP-1 and -2 gene expression, but decreased IGFBP-3 gene expression, while in thyrotoxicosis there are normal IGFBP-1 and -2 mRNA levels but increased IGFBP-3 gene expression. These results suggest that there is specific and different transcriptional regulation for IGFBP-1, -2 and -3 in hypoand hyperthyroid rats.
Journal of Endocrinology (1994) 140, 251–255
Influence of ovarian status and regional fat distribution on protein kinase C in rat fat cells
Authors: Lacasa D , Agli B , Mur M , Dausse J-P , Giudicelli Y 
Published: December 2008

Abstract
The effects of ovarian status on insulin- and phorbol 12-myristate 13-acetate (TPA)-stimulated lipogenic responses, phorbol ester-specific binding to protein kinase C (PKC) and immunoblot-quantified β- and ε-PKC isoform levels were compared in female rat fat cells from subcutaneous and deep intra-abdominal (parametrial) fat deposits.
In control rats, the cytosolic PKC content per cell was 70% lower in subcutaneous than in parametrial adipocytes. In subcutaneous and parametrial fat cells, the cytosolic PKC contents were reduced by ovariectomy and restored to normal by the administration of ovarian hormones (oestradiol plus progesterone). However, the lipogenic response to TPA was unaltered by ovarian status in both fat deposits, contrasting with the insulin-stimulated lipogenic response. This response increased in parametrial fat cells after ovariectomy and returned to normal after ovarian hormone treatment whereas, in subcutaneous fat cells, the insulin response was either unaltered by ovariectomy or increased by the administration of ovarian hormones. This study shows important site-related differences in fat cell PKC content but also reveals a similar modulation of this PKC by ovarian status, regardless of the anatomical localization of the fat cells. The physiological significance of these findings is discussed.
Journal of Endocrinology (1994) 140, 275–282
Inhibition of aromatase activity in rat Sertoli cells by thyroid hormone
Authors: Ulisse S , Jannini E A , Carosa E , Piersanti D , Graziano F M , D'Armiento M 
Published: December 2008

Abstract
Basal and FSH-induced aromatase activity in prepubertal rat Sertoli cells was inhibited by l-tri-iodothyronine (T3) in a time- and dose-dependent manner. The effect was evident only after 6 h of preincubation with T3 (10−7 m) and the half-maximal dose was 0·5 ±0·2 nm, which correlated with the Kd of the nuclear T3 receptor of rat Sertoli cells (Kd=1–2 nm). The effect was specific as judged by the lack of effect of the T3 analogue 3-iodo-l-thyrosine. The inhibitory effect of T3 was present over the entire range of FSH concentrations used (0·001–100 ng/ml). In T3-treated Sertoli cells, aromatase activity induced by 8-bromo-cyclic AMP was inhibited by the same order of magnitude as that of FSH, thus suggesting that the inhibitory effect of T3 was downstream from cyclic AMP formation. Furthermore, pretreatment of Sertoli cells cultures with T3 (24 h, 10−7 m) did not affect basal or FSH-induced extracellular cyclic AMP accumulation. This effect of T3 on rat Sertoli cell aromatase activity may be regarded as a part of the integrated mechanism by which thyroid hormone modulates the functions of the seminiferous epithelium.
Journal of Endocrinology (1994) 140, 431–436
Polymorphism of human pituitary FSH: analysis of immunoreactivity and in vitro bioactivity of different molecular species
Authors: Simoni M , Jockenhövel F , Nieschlag E 
Published: December 2008

Abstract
Follicle-stimulating hormone is known to be highly heterogeneous in serum and in the pituitary. In the present study, we have partially separated different molecular species of human pituitary FSH and characterized their immunoreactivity and in vitro bioactivity. Pooled extracts of male (n=15) and female (n=9) human pituitary glands were chromatographed on a column of Sephacryl S-200 and FSH-containing fractions were fractionated by chromatofocusing in the pH range 4–6. FSH was measured in the individual fractions by an in vitro bioassay, based on the FSH-dependent aromatase activity of immature rat Sertoli cells, and by the following methods based on commercial kits: radioimmunoassay (RIA), immunofluorimetric assay (IFMA), immunoradiometric assay (IRMA), immunoenzymometric assay (IEMA). In each assay, the kit standard, calibrated against the 2nd International Reference Preparation (IRP) 78/549, and the International Standard (IS) 83/575 were run in parallel. The relative potencies of the kit standards in terms of IS 83/575 were: IFMA 3·08, IRMA 1·62, RIA 2·42, IEMA 1·45 and bioassay 1·14. After chromatofocusing, pituitary FSH eluted mostly in fractions with pH≈4·5, without sex-related differences. In both sexes ≈25% of bioactive material showed a pI<4 and eluted with 1 m NaCl. Although the same IS 83/575 was used in the various assays, the profiles of immunoreactive FSH were significantly different. The highest intermethod variability was observed in the case of male pituitary FSH. The relative biopotency of the different molecular species of FSH did not appear to change according to their pI but, rather, varied with the assay method and the standard. In terms of 2nd IRP 78/549 the activity profiles were similar but not identical to those obtained in terms of IS 83/575 and the ratios between the two values (IS:IRP ratios) were significantly different among the methods. No significant correlation was found between IS:IRP ratios and FSH concentrations or pH. These data suggest that: (1) all the methods have different affinities for standard and unknown and/or for different molecular isoforms of FSH; (2) overall, they perform more accurately when assaying female pituitary FSH, perhaps because of a closer resemblance between standard and unknown; (3) the variability of the IS:IRP ratios indicates a different affinity of the antibodies for IS 83/575 and the kit standard, highlighting the importance of the molecular composition of the reference preparations for the final result; and (4) the results do not support the commonly accepted concept of a higher in vitro biopotency of less acidic species of pituitary FSH.
Journal of Endocrinology (1994) 141, 359–367
Growth hormone (GH) regulation of circulating insulin-like growth factor-I levels during sexual maturation of the GH-deficient dwarf (dw/dw) male rat
Authors: Crawford B A , Dobbie P , Bass J J , Lewitt M S , Baxter R C , Handelsman D J 
Published: December 2008

Abstract
In many mammalian species, circulating levels of insulinlike growth factor-I (IGF-I) rise during puberty. Previous studies manipulating testosterone levels in rats with normal GH secretion suggested that the pubertal IGF-I rise is regulated by an interaction between GH and sex steroids. Therefore, in a reciprocal study, IGF-I levels were examined during sexual maturation of the GH-deficient dwarf (dw/dw) rat which has a selective genetic deficiency of GH but normal sex steroid levels. Male dw/dw rats were treated with daily injections of recombinant human GH (200 μg/100 g body weight) or saline vehicle, from 28 to 70 days of age. Sexual maturation was determined to occur primarily between 42 and 63 days of age based on testis and seminal vesicle growth and plasma testosterone levels. GH treatment had no effect on seminal vesicle weights, plasma testosterone or gonadotrophins. GH administration resulted in a 7% increase in absolute testes weight (P<0·05), but a 50% increase in body weight (P<0·0001). These results supported previous findings that the reproductive development of dw/dw rats is essentially normal. Untreated dw/dw rats had no rise in IGF-I levels during sexual maturation. In contrast, treatment with GH produced a marked sustained rise in IGF-I levels (P<0·0001). Ligand blots demonstrated GH induction of IGF-binding protein-3 (IGFBP-3) and an IGFBP cluster at 32 kDa. The initially high immunoreactive IGFBP-1 levels (>600 ng/ml) decreased by 49 days of age after which untreated dw/dw rats had significantly higher IGFBP-1 levels than GH-treated dw/dw rats (P<0·01). We conclude that GH secretion, rather than sex steroids, may be the predominant determinant of pubertal IGF-I levels in rats and that the rise in circulating IGF-I levels during puberty is not an indispensible event for normal reproductive development.
Journal of Endocrinology (1994) 141, 393–401
Annual variations in serum thyroid-stimulating hormone and thyroid hormones and in their responses to thyrotrophin-releasing hormone in the reindeer
Authors: Timisjärvi J , Ojutkangas V , Eloranta E , Nieminen M , Leppäluoto J , Liimatainen S , Vuolteenaho O 
Published: December 2008

Abstract
The reindeer in its natural habitat is subject to great annual variations in ambient temperature, illumination and nutrition. To ascertain the effect of these environmental factors on thyroid function, serum thyroid-stimulating hormone (TSH), thyroxine (T4), tri-iodothyronine (T3) and reverse T3 (rT3) concentrations were measured four times a year (2 June, 8 October, 21 November, and 24 February) in 14 animals housed outdoors at latitude 69°10′N. They all showed statistically significant (P<0·05) seasonal changes. Serum TSH and T4 were highest in February (623 ± 30 ng/ml and 287 ± 19 nmol/l respectively). TSH was lowest in October (318 ± 47 ng/ml) and T4 in November (199 ± 19 nmol/l). The T3 concentration was highest in November (3·0 ± 0·3 nmol/l) and lowest in June (1·8 ± 0·2 nmol/l). In contrast, rT3 was highest in June (3·6 ± 1·2 nmol/l) and lowest in November (1·9 ± 0·6 nmol/l). Thus, there was an inverse relationship between T3 and rT3 (linear regression r= −0·406, P<0·01).
TSH, T4, T3 and rT3 responses to exogenous thyrotrophin-releasing hormone (synthetic TRH; 500 μg i.m.) were determined in ten animals. The magnitude of their response to TRH was significantly (P<0·05) dependent on the time of year. When compared with the control level all the parameters rose significantly (P<0·05). The greatest rise in serum TSH occurred in October (219 ± 151% and the smallest in February (66 ± 53%). The corresponding figures for T4 were 56 ± 60% in June and 11 ± 18% in February, for T3 143 ± 45% in October and 73 ± 46% in June, and for rT3 25 ± 13% in October and 12 ± 10% in February.
In conclusion, significant annual variations occur in the reindeer serum concentrations of TSH and thyroid hormones as well as in their responses to exogenous TRH. Environmental factors greatly affect the pituitary-thyroid axis in this mammalian species living far in the North. The observed variations probably reflect an adaptation of the species to the annual environmental cycle.
Journal of Endocrinology (1994) 141, 527–533
Physiological roles of thyrotrophin-releasing hormone and vasoactive intestinal peptide on the pulsatile secretory patterns of prolactin in pituitary-grafted female rats
Authors: Lafuente A , Marcó J , Esquifino A I 
Published: December 2008

Abstract
Much is known about the fact that thyrotrophin-releasing hormone (TRH) and vasoactive intestinal peptide (VIP) stimulate prolactin secretion but areas of uncertainty remain. This work was undertaken to describe the effects of TRH and VIP on the pulsatile secretion pattern of prolactin, in adult sham-operated and pituitary-grafted hyperprolactinaemic female rats. Two pulses of TRH (1 μg/rat) or one pulse of VIP (20 μg/rat) were given 60 or 120 min after the period of blood sampling. Pituitary grafting increased the mean values of prolactin, absolute amplitude and duration of the peaks and decreased their frequency, compared with control animals. In sham-operated rats, TRH elevated prolactin levels by increasing the absolute and relative amplitudes and duration of the pulses, along with a decrease in their frequency. No priming effects of TRH were observed in this study. Hyperprolactinaemia blunted TRH effects on the pulsatile secretion pattern of prolactin. In sham-operated rats, VIP administration increased the absolute and relative amplitudes of the prolactin peaks. None of the other parameters studied were changed. In pituitary-grafted animals, VIP administration increased the absolute and relative amplitudes of the prolactin peaks but to a lesser extent compared with controls. These data suggest that TRH and VIP affect prolactin pulsatility differentially. The effects of TRH and VIP were blunted to some extent by exposure to previously elevated circulating prolactin levels.
Journal of Endocrinology (1994) 142, 581–586
Selective inhibition of mouse placental lactogen II secretion by tumour necrosis factor-α
Authors: Yamaguchi M , Ogren L , Barnard R , Imai T , Sawada T , Miyake A , Talamantes F 
Published: December 2008

Abstract
The placental members of the prolactin-GH-placental lactogen (PL) gene family of the mouse include mPL-I, mPL-II, proliferin (PLF) and proliferin-related protein (PRP). The aim of the present study was to assess the effects of tumour necrosis factor-α (TNF-α) on the secretion of these proteins in primary cultures of placental cells from days 7, 9 and 12 of pregnancy. The effects of epidermal growth factor (EGF) on the secretion of PLF and PRP were also determined. EGF has previously been shown to stimulate mPL-I and inhibit mPL-II secretion. Incubation of placental cells from day 7 of pregnancy for 5 days with 10 nmol human (h)TNF-α/1 did not affect the mPL-II concentration of the medium, but similar treatment of cells from days 9 or 12 of pregnancy resulted in a significant reduction in the mPL-II concentration of the medium by the second or third day of culture. The intracellular concentration of mPL-II, the number of cells that released mPL-II as assessed by reverse haemolytic plaque assay, and steady-state levels of mPL-II mRNA as assessed by Northern analysis were also reduced by hTNF-α treatment. The lowest concentration of hTNF-α that significantly inhibited mPL-II secretion by cells from day 12 of pregnancy was 0·01 nmol/l. hTNF-α treatment did not affect the secretion of mPL-I, PLF or PRP, as assessed by the concentrations of these proteins in the medium during a 5-day incubation. Incubation of the cells with 20 ng EGF/ml also did not affect the PLF or PRP concentration of the medium during 5 days of culture. To determine whether the effect of hTNF-α on mPL-II secretion was mediated by interleukin-6 (IL-6), the IL-6 concentration of the medium of control and hTNF-α-treated cells was determined. Bioactive and immuno-reactive IL-6 could not be detected in medium from either treatment group. The presence of binding sites for hTNF-α was assessed in cells from day 12 of pregnancy. Scatchard analysis detected a single class of binding sites having a Kd of 1·61±0·34 nmol/l, with about 1350 sites per cell. The results of this study demonstrate that hTNF-α inhibits the secretion of mPL-II by placental cells from days 9 and 12 of pregnancy, suggesting that TNF-α may be one of the factors that regulate the production of this hormone in vivo.
Journal of Endocrinology (1994) 143, 95–105
Regulation of oxytocin production by purified adult rat Leydig cells in vitro: effects of LH, testosterone and lipoproteins
Authors: Frayne J , Nicholson H D 
Published: December 2008

Abstract
The aim of the present study was to determine whether LH stimulates oxytocin production by adult rat Leydig cells directly or indirectly via testosterone. Purified adult rat Leydig cells were cultured in the presence or absence of 0·1 ng/ml LH or 1, 10 or 100 ng/ml testosterone for 22 h. Culture medium was collected at 2-hourly intervals and assayed for oxytocin and testosterone. In the presence of LH, Leydig cells produced significantly higher levels of both testosterone (basal production 1·4± 0·13 ng, LH-stimulated 4·1 ±0·13 ng/106 cells per 2 h) and oxytocin (basal production 8·3± 1·2 pg, LH-stimulated 20·2± 1·3 pg/106 cells per 2 h). Testosterone also stimulated oxytocin secretion. However, the increase was smaller compared with that seen with LH and was not found to be dose-dependent. Furthermore, testosterone production was only significantly increased by LH during the first 10 h of the 22-h culture period whereas LH stimulated oxytocin production throughout the whole culture period.
To further determine the effect of LH on oxytocin production, cultures were performed in the presence of LH and/or 400 μm aminoglutethimide. In the presence of aminoglutethimide both the basal and LH-stimulated production of testosterone was significantly reduced. However, in the same cultures aminoglutethimide did not alter either the basal or LH-stimulated production of oxytocin.
These data show that LH does not act via testosterone to stimulate oxytocin production and therefore acts directly or by some alternative indirect mechanism.
In this study it was found that two other factors, cell density and lipoprotein, also influenced oxytocin production by isolated Leydig cells. Decreasing the density at which Leydig cells were cultured from 106 to 10 cells/well significantly increased both their basal and LH-stimulated production of oxytocin. Lipoproteins were also found to stimulate oxytocin production in a dose-dependent manner and to synergize with LH to further increase LH-stimulated oxytocin production. The results of this study show the production of oxytocin by Leydig cells to be regulated not only by the gonadotrophin LH but also by lipoproteins which are known to be present in interstitial fluid. These data add to the accumulating evidence that intratesticular oxytocin may be a paracrine factor.
Journal of Endocrinology (1994) 143, 325–332
Induction of an acute phase response in lambs causes an increase in plasma levels of GH and IGF-I
Authors: Moore L G , Pfeffer A , Ng Chie W , Miller H A , Rogers K M , O'Keeffe L E 
Published: December 2008

Abstract
GH and IGF-I plasma concentrations were measured in lambs during an acute phase response induced by an intrathoracic injection of yeast. The acute phase response was indicated by reduced feed intake, weight loss and an increase in plasma concentrations of the acute phase protein haptoglobin. Intensive blood sampling on day 1 revealed elevated basal concentrations of GH in the yeast-injected group compared with concentrations in pair weight and ad libitum fed control lambs. This suggests that at the beginning of an acute phase response there is an increase in either GH secretion or the half life of GH. No evidence of a specific GH-binding protein in sheep plasma could be detected. IGF-I concentrations in the yeastinjected group remained constant for 3 days then increased to a peak level at day 6. In contrast, plasma IGF-I concentrations were depressed from days 3 to 6 in the pair weight control group and they were unchanged in the ad libitum fed controls. When the IGF-I concentrations were elevated in the yeast-injected group, this group had a higher daily weight gain despite their lower feed intake compared with the ad libitum fed controls. These results suggest that IGF-I may be associated with the increase in weight in the late stage of an acute phase response during recovery from an infection or injury. Day 1 GH peak amplitude concentrations in the yeast-injected lambs were negatively correlated with IGF-I concentrations on the following 2 days yet in the pair weight lambs the correlation was in a positive direction suggesting that the relationship between GH and IGF-I is different between animals that lose weight during an acute phase response and animals that lose weight because of feed restriction.
Journal of Endocrinology (1995) 144, 243–250