Subcutaneous implantable infusion device for chronic intravenous pulsatile gonadotropin-releasing hormone treatment

Preliminary data indicate the potential utility of an implantable subcutaneous device that facilitates chronic intravenous infusion of pulsatile gonadotropin-releasing hormone (GnRH) for ovulation induction. GnRH distribution curves were congruent in control monkeys and those with implanted devices. Tissue tolerance was good in this brief trial. These findings suggest that use of this or a similar implantable device be considered for chronic GnRH administration in human pulse therapy.     

Utilization of the Synthetic Phosphagen Cyclocreatine Phosphate by a Simple Brain Model During Stimulation by Neuroexcitatory Amino Acids

The ability of 1-carboxymethyl-2-imino-3-phosphonoimidazolidine (cyclocreatine-P), accumulated by a simple brain model, to function as a supplemental synthetic phosphagen and respond to the decreases in cytosolic ATP/free ADP ratios that occur during prolonged stimulation by various excitatory amino acids was investigated. Suspensions of chopped whole brain from 11- to 14-day-old chick embryos were incubated with 30 mM cyclocreatine for 90 min, resulting in accumulation of 100 mumol/g dry weight of cyclocreatine-P, and then incubated for up to 1 h with a series of excitatory amino acids of widely differing potencies. Under these conditions net utilization of cyclocreatine-P was detected in response to stimulation by the following neuroexcitatory compounds at the indicated threshold concentrations: kainate (20 microM), N-methyl-DL-aspartate (20 microM), L-homocysteate (20 microM), L-glutamate (200 microM), D-glutamate (200 microM), L-aspartate (2 mM), DL-2-amino-3-phosphonopropionate (2 mM), and DL-2-amino-4-phosphonobutyrate (2 mM). Significant increases in water content of chick embryo brain minces accompanied stimulation by excitatory amino acids. It is suggested that changes in water content or cyclocreatine-P levels in this sensitive brain model might be utilized in automatable screening procedures for detecting novel antagonists and/or new agonists of excitatory amino acids.     

Distribution of salmon gonadotrophin releasing-hormone in the brain and pituitary of the sea bass (Dicentrarchus labrax)

Summary The distribution of salmon gonadotrophin-releasing hormone (sGnRH) was studied in the brain and pituitary of two-year-old immature sea bass (Dicentrarchus labrax) by means of an enzymoimmunoassay (EIA) for sGnRH and immunocytochemistry. The EIA for sGnRH is a competitive assay using a tracer made of sGnRH coupled to acetylcholinesterase from an electric eel. The separation of free and bound tracer is achieved by coating the plates with mouse anti-rabbit IgG monoclonal antibodies. Displacement curves generated by sGnRH and extracts from pituitary and different brain regions showed a good parallelism allowing the assay to be used for sGnRH measurements in this species. Although all parts of the brain contained measurable levels of sGnRH, the highest concentrations were found in the pituitary, the olfactory bulbs and the telencephalon. These data were confirmed by immunocytochemistry. Cell bodies were found in the olfactory bulbs, ventral telencephalon, preoptic region and mediobasal hypothalamus. Immunoreactive fibers could be observed in all parts of the brain including the optic tectum, the cerebellum (corpus and valvula), the vagal lobe, the medulla oblongata and the rostral spinal cord. In most cases, these fibers do not form well defined bundles; however, there was clearly a continuum of immunoreactive fibers, extending from the olfactory bulbs to the pituitary, and along which all the cell bodies described above were located. In the ventral telencephalon and the preoptic region, clear pictures of varicose positive fibers contacting immunoreactive perikarya could be observed. These data indicate that sGnRH is most likely an endogenous peptide in the brain of the sea bass, although the presence of other forms of GnRH cannot be excluded at this point. This study also demonstrates that the general organization of the GnRH systems in the sea bass is highly similar to what has been described in most freshwater teleost species, and provides basis for further studies on the neuroendocrine control of gonadotrophin release in this commercially important species.     

Effects of chronic treatment with a GnRH agonist on oestrous behaviour and on the secretion of LH and progesterone in the ewe

A study was carried out to investigate a novel approach to oestrus synchronization in the ewe by treatment with a gonadotrophin releasing hormone (GnRH) agonist. Groups of ewes were initially treated on Day 2, 10 or 14 of the oestrous cycle with 10 mug GnRH analogue (D-Ser(Bu(t)) 6 des Gly GnRH ethylamide) per ewe per day for 14 days. Behavioural oestrus was inhibited during GnRH agonist treatment and recurred from 8 to 38 days after the treatment in an unsynchronized manner. Luteal activity during treatment was not impaired but reduced progesterone concentrations occurred in cycles after the treatment. The rhythm of ovarian function, generally characterized by prolonged follicular development, was impaired. During the treatment and subsequent recovery period, integrity of pituitary function was examined by measuring luteinizing hormone (LH) after GnRH agonist was injected, and after stimulation test doses of 150 ng natural GnRH were administered. During treatment there was, with time, a decline in pituitary response to the agonist which suggested that pituitary release of LH was exhausted. After the 14-day treatment the stimulation test with GnRH revealed a gradual return to normal responsiveness although this was not complete three weeks after the treatment when compared to control ewes. This lowered pituitary activity could cause the impaired ovarian function.     

Effects of N,N′-dicyclohexylcarbodiimide on isolated and reconstituted cytochrome b-c1 complex from bovine heart mitochondria

N,N′-Dicyclohexylcarbodiimide (DCCD) inhibits the activity of ubiquinol-cytochrome c reductase in the isolated and reconstitued mitochondrial cytochrome b-c₁ complex. DCCD inhibits equally electron flow and proton translocation (i.e., the $\text{H}{}^{\mathrm{+}}\text{e}{}^{\mathrm{?}}$ ratio is not affected) catalysed by the enzyme reconstituted into phospholipid vesicles. The inhibitory effects are accompanied by structural alterations in the polypeptide pattern of both isolated and reconstituted enzyme. Cross-linking was observed between subunits V (iron-sulfur protein) and VII, indicating that these polypeptides are in close proximity. A clear correlation was found between the kinetics of inhibition of enzymic activity and the cross-linking, suggesting that the two phenomena may be coupled. Binding of [¹⁴C]DCCD was also observed, to all subunits with the isolated enzyme and preferentially to cytochrome b with the reconstituted vesicles; in both cases, however, it was not correlated kinetically with the inhibition of the enzymic activity.     

Synthesis of methyl

利用放射免疫分析法测定了排卵、排精前后青岛文昌鱼体内促性腺激素释放激素（ＧｎＲＨ）的含量变化,并通过高效液相色谱比较了雌、雄文昌鱼性腺及除性腺外体部ＧｎＲＨ的种类和含量的异同。结果表明：１）生殖过程中雌、雄文昌鱼体内ＧｎＲＨ含量的变化规律不同;雌性文昌鱼体内ＧｎＲＨ总含量在排卵时有所下降,排后１２小时政策最为明显,以后逐渐上升到排前水平;雄性文昌鱼仅在排精时有所下降,２小时后即稳定在排水平。２）文     

排卵,排精前后文昌鱼体内GnRH的研究

利用放射免疫分析法测定了排卵、排精前后青岛文昌鱼体内促性腺激素释放激素（ＧｎＲＨ）的含量变化,并通过高效液相色谱比较了雌、雄文昌鱼性腺及除性腺外体部ＧｎＲＨ的种类和含量的异同。结果表明：１）生殖过程中雌、雄文昌鱼体内ＧｎＲＨ含量的变化规律不同;雌性文昌鱼体内ＧｎＲＨ总含量在排卵时有所下降,排后１２小时政策最为明显,以后逐渐上升到排前水平;雄性文昌鱼仅在排精时有所下降,２小时后即稳定在排水平。２）文     

Relationship between osmoprotection and the structure and intracellular accumulation of betaines by Escherichia coli

Abstract Naturally occuring betaines, especially glycine betaine and proline betaine, were accumulated by Escherichia coli from urine. In synthetic hyperosmotic medium, with an homologous series of added betaines, (CH₃)₃N⁺-(CH₂) _n -COO⁻, osmoprotective activity and intracellular accumulation decreased monotonically as n increased from 1 to 5. In contrast, α -substituted glycine betaines were accumulated in a similar manner to glycine betaine, but with different osmoprotective activities. Arsenobetaine, with a quaternary arsonium group, was also accumulated but amino acids which can become negatively charged in a chemically basic environment were not.     

Leuprolide acetate can reversibly prevent egg laying in cockatiels (Nymphicus hollandicus)

Leuprolide acetate acts as a superactive gonadotropin-releasing hormone (GnRH) agonist in mammals. Its administration to humans in a depot formulation (Lupron Depot 3.75 mg; TAP Pharmaceuticals, Deerfield, IL) consisting of microspheres suspended in a diluent of carboxymethylcellulose and other elements leads to an initial increase in serum gonadotropin levels followed by a prolonged suppression of ～ 1 month's duration. To test whether it might act in cockatiels to prevent egg laying, we administered Lupron Depot to groups of pairs (at least 6 pairs/group) stimulated to reproduce by provision of nest boxes and exposure to sexually stimulatory daylengths (15:9 L:D). A single intramuscular injection of Lupron Depot, calculated to achieve a daily release rate of 0 (control; diluent only), 17, 52, or 156 μg/kg/day of leuprolide acetate, was administered on day 0, when birds received nest boxes and after daylength had been stepwise increased. Egg production began on day 12 in the 0 and 17 μg dose groups, and on day 31 in the groups receiving the higher doses. Number of eggs per clutch, candled fertility, and percent hatchability were not significantly different among the groups. In a separate experiment in which leuprolide was administered prior to photostimulation and nest box presentation, nest-inspection behavior was not prevented. We conclude that a single injection of Lupron Depot is effective in reversibly preventing egg laying in cockatiels. © 1994 Wiley-Liss, Inc.     

Morpho-functional aspects of the hypothalanus-pituitary-gonadal axis of elasmobranch fishes

Synopsis The tetrapod hypothalamus-pars distalis axis contains a blood portal system. Contrarily, elasmobranchs appear to lack a direct vascular supply from the hypothalamus to the ventral lobe of the pituitary where gonadotropic activity resides. The hypothalamus contains GnRH immunoreactivity and GnRH causes an increase in plasma gonadal steroids, perhaps via ventral lobe stimulation. Therefore, the question arises as to how GnRH reaches the pituitary. We suggest that the general circulation route might be practicable. Indeed, in the plasma of the electric ray,Torpedo marmorata, a major early eluting form has been detected using high performance liquid chromatography coupled with region specific radioimmunoassay. The presence of GnRH in the blood may allow the molecule to reach the gonads and to act there by direct mechanisms. Intragonadal levels of steroids may have a paracrine and/or autocrine role in the regulation of steroidogenesis in the testis and in the development f specific germinal cell stages. Particularly, the zonated morphology of the testis supports the concept of a diverse environment for different spermatogenic stages. Finally, gonadal steroids may feed back to affect pituitary activity.