Somatic growth in hypophysectomized pituitary-homografted rats is promoted by prolactin

Summary Hypophysectomized male rats bearing a homograft of two adenopituitaries under the kidney capsule showed a significant increase in b.wt as compared to hypophysectomized non-homografted animals. Radiommunoassay of growth hormone (GH), ACTH, -MSH,-endorphin and prolactin (PRL) revealed that only the latter was highly increased in the plasma of hypophysectomized homografted rats. These animals showed also increased levels of plasma corticosterone. However, daily injection of corticosterone failed to promote somatic growth in hypophysectomized non-homografted rats. These results suggest that PRL, and not other hormonal factors, promotes somatic growth in hypophysectomized homografted rats, and stress the concept that only PRL is secreted in significant amounts by pituitary homografts.     

Self-labeling of human polymorphonuclear leucocyte myeloperoxidase with125iodine

In order to obtain a radioimmunoassay (RIA) technique for the measurement of human plasma myeloperoxidase (MPO), we purified the enzyme from polymorphonuclear granulocytes (neutrophils), and compared three methods of labeling it with¹²⁵Iodine: chloramine T, lactoperoxidase, and an original technique of self labeling based on the ability of the enzyme to oxidize and bind¹²⁵I in the presence of H₂O₂. The chloramine T technique produced a degraded protein, as well shown by a high non-specific binding of tracer to antibody. The lactoperoxidase technique did not succeed in labeling MPO with an adequate specific activity. In contrast, the self-labeling method gave a stable tracer with a specific activity of 23 Ci/gmg MPO (85 MBq), a satisfactory level of immunoreactivity, and a low-specific binding (3%). After labeling, purification of tracer was achieved by gel filtration chromatography in phosphate buffer (0.05 M; pH7) to which 0.1% poly-L-lysine was added. The labeled molecule remained stable for 40 days and could be used for RIA with a polyclonal antibody raised in rabbits.     

Occurrence of 3-epi-22-deoxy-20-hydroxyecdysone and its phosphoric ester in diapause eggs of the silkworm,Bombyx mori

Ecdysteroids in diapause eggs of the silkworm,Bombyx mori, were analyzed using high-performance liquid chromatography (HPLC) combined with radioimmunoassay (RIA). A relatively large amount of an unidentified free ecdysteroid and its phosphoric ester (conjugated form) were detected. These two compounds were isolated by a combination of column chromatography on silicic acid, thin-layer chromatography (TLC), and HPLC using a reverse-phase (RP) column. The purified compounds were identified as 3-epi-22-deoxy-20-hydroxyecdysone (22d20E) and 3-epi-22-deoxy-20-hydroxyecdysone 2-phosphate (22d20E2P) by means of mass spectrometry and nuclear magnetic resonance spectroscopy. to our knowledge, this is the first report of 22d20E and 22d20E2P.     

Reduced tuberoinfundibular dopaminergic neuronal function in rats after long-term withdrawal of estrogen treatment

Summary Hypothalamic fragments from female rats treated repeatedly with estradiol valerate (EV) and bearing prolactin (PRL)-secreting tumors contained, seven months after the last EV injection, lower concentrations of dopamine (DA) than age-matched controls. Depolarizing concentrations of K⁺ (35 mM) and amphetamine (50 M) evoked in PRL-secreting tumor bearing rats an endogenous DA release significantly lower than in controls.     

Analysis of translation products synthesized in isolated rat hepatocytes treated with diethylnitrosamine

Summary Isolated rat hepatocytes were labeled with³⁵S-methionine in the presence of 25 mM diethylnitrosamine (DENA). The intrinsically labeled proteins were analyzed by one-and two-dimensional gel electrophoresis and the fluorographic patterns were compared with those obtained from untreated hepatocytes. The results of short term experiments (2 h) show that, in the presence of 25 mM DENA, protein synthesis is inhibited by 50%. This reduction encompasses all protein species without selective inhibition of certain proteins.This work was supported by CNR (Project Control of Neoplastic Growth) grant No. 810132696 and partially by AIRC.     

Changes in ecdysone levels in the spiderPisaura mirabilis nymphs (Araneae,Pisauridae)

Summary RIA of ecdysones are made on nymphal extracts ofPisaura mirabilis during the intermoulting cycle. The hormone level is low during exuviation and the greatest part of the intermoulting period but increases notably before and after ecdysis. The main hormonal peak or pre-ecdysial peak is due to -ecdysone, characterized by combined TLC/RIA.Acknowledgment. We wish to thank Dr M. Hirn for his help during the manipulations and J. Britton who has translated the text into English.     

Respective roles of circulating T4 and T3 in control of TSH secretion in severely iodide-deficient rats

Summary After a 6-month iodide deficiency, Wistar male rats were submitted to a normal iodine diet (20 and 50 g of¹²⁷I daily). Plasma T₃, T₄ and TSH were determined by RIA from 0 to 140 days of iodide refeeding. A highly significant correlation was found between plasma TSH and T₄ concentrations, but not between plasma TSH and T₃ levels. These data suggest that an increase in plasma T₃ alone, up to the normal value, is not able to inhibit TSH secretion. It is only when a certain plasma T₄ concentration is also reached, resulting in further T₃ formation through deiodination, that TSH secretion is inhibited.Dedicated to the memory of Prof. C. Simon.This work was supported by grants from the CNRS (Equipe de Recherche Associée no 234) and from the INSERM (A.T.P. 49.77.8).Acknowledgments. The authors wish to thank Mrs M. Chartier for valuable technical assistance. They are indebted to the rat pituitary distribution programme of NIAMDD, NIH, Bethesda, for their gift of rat TSH reagents.     

The effect of camphor on mitochondrial respiration

Summary Camphor at <8 moles/mg protein reduced the rate of oxygen consumption by rat liver mitochondria. The effect occurs only with NAD⁺-linked substrates. Succinate linked respiration was inhibited but this appears to be caused by some conversion of succinate to malate. At higher levels, camphor increases oxygen consumption with succinate substrate, by uncoupling at site II.Acknowledgment. D.F.G.-C. is grateful to the New Zealand Cancer Society (Wellington Branch), for financial support. We thank Mrs E. Dye for technical assistance and Dr W. Jordan for valuable discussions.     

Involvement of H1 receptors in the central antinociceptive effect of histamine: Pharmacological dissection by electrophysiological analysis

Intracerebroventricular (i.c.v.) administration of histamine (HA, 0.025–0.1 M/rat) to arthritic rats induces a dose-related inhibition of the neuronal thalamic firing evoked by peripheral noxious stimuli. To characterize the type(s) of HA receptors involved in this depressing activity of the amine we used electrophysiological techniques to examine the effects of i.c.v. administration of H₁ and H₂ agonists and antagonists on the spontaneous and evoked nociceptive firing of the thalamic neurons in rats rendered arthritic by Freund's adjuvant. The H₁ agonist 2-pyridylethylamine (0.4–1.0 M/rat, i.c.v) displayed a dose-dependent antinociceptive effect very similar to that of HA, while the H₂ agonist dimaprit (0.05–0.2 M/rat, i.c.v.) did not modify thalamic firing. Neither mepyramine (H₁ antagonist, 0.1 M/rat, i.c.v.) nor zolantidine (H₂ antagonist, 0.01 M/rat, i.c.v.) modified the evoked firing of rat thalamic neurons. When administered before HA (0.1 M/rat, i.c.v.) mepyramine but not zolantidine was able to inhibit the antinociceptive effect of HA. On the basis of the present electrophysiological results, we suggest that a specific interaction of histamine with H₁ receptors may be important for its antinociceptive effect on afferent peripheral inputs to the thalamus.     

Age-related differences in the urinary excretion of prostaglandin F2α catabolites in the rat

Summary Tritium-labelled PGF₂ was administered i.v. into rats of varying ages (2, 4, 6 weeks and adult). Urine was collected and assayed for radioactive products by thin-layer-chromatography. Results showed a distinctly different urinary profile between the 2-week-old and the adult rat. While the urinary pattern from the 2-week-old rat gave a single less polar product than PGF₂, the pattern from the adult rat gave products more polar than PGF₂. Urine from the 4- and 6-week-old rats gave a mixture of these types of products. These results indicate that some prostaglandin catabolic pathway (likely the -oxidative system) is activated in vivo within the 4–6-week postnatal period in the rat.Supported by a grant (MT-4181) to C.P.-A. from the Medical Research Council of Canada.This study is in partial fulfillment of the requirements for a Ph.D. degree in the Department of Pharmacology, University of Toronto.     

Expression of the mutant gene for L-gulono-γ-lactone oxidase in scurvy-prone rats

A mutant strain of Wistar rats with L-gulono--lactone oxidase deficiency has recently been established. To investigate this deficiency by DNA and RNA blot hybridization analyses, a fragment of a previously cloned cDNA encoding rat L-gulono--lactone oxidase was used as a probe. When genomic DNA of the mutant rat was digested with several restriction enzymes, the probe hybridized to fragments of the same sizes as those produced from DNA of normal rats. Poly(A)⁺RNA from the liver of the mutant rat was found to contain an L-gulono--lactone oxidase-specific mRNA of a normal size at a comparable level to that of normal rats. An in vitro translation experiment revealed that the mRNA programmed the synthesis of an enzyme protein which had the same molecular weight as that of the translational product of the normal mRNA, although the amount synthesized was markedly reduced as compared with that synthesized with the normal mRNA. In accordance with this observation, a very low but definite degree of L-gulono--lactone oxidase activity was detected in the microsomes of the mutant rat by a newly developed, highly sensitive method.Acknowledgments. The authors thank Dr Susumu Makino, Shionogi Research Laboratories, Shionogi & Co., Ltd, Japan, for his kind donation of normal (ODS- +/+) and ODS (ODS-od/od) rats. This work was supported in part by Grant-in-Aid (59570103) for Scientific Research from the Ministry of Education, Science and Culture of Japan.     

A comparative study of membrane related phenomena in normal and crown gall tissues of red beet (Beta vulgaris L.)

Summary Divalent metal ions have been found to protect membranes of red beet crown gall tissue more than their adjacent normal regions from thermal or gamma radiation stress, suggesting the possibility of an alteration on the surface charge accompanying tumor formation in plants. Further, tumor tissue has been observed to possess enhanced membrane ATPase activity, a higher tissue sulfhydryl content and increased protein levels, thus suggesting a model of source (normal tissue) and sink (tumor tissue) relationship.Acknowledgments. Thanks are due to Dr Sudhir Sopory, School of Life Sciences, Jawaharlal Nehru University, New Delhi for a critical discussion on this paper. KVAR thanks the Council of Scientific and Industrial Research, New Delhi, for financial assistance in the form of a Senior Research Fellowship.     

Phytosterols from pressmud residue obtained after methanogenic fermentation

Summary Pressmud, a sugar factory waste, was fermented with methanogenic bacteria in an anaerobic fermenter for 40 days at 31±2°C. The pressmud residue obtained after fermentation was used as a source for the extraction of phytosterols. The anaerobic digestion degraded the organic matter and resulted in enrichment of phytosterols from 0.33% in the pressmud to 3.05% in the residue. Refluxing of 100 g of residue with benzene, petroleum ether, and ethanolic KOH (1051) yielded 8 g of soft cake, which on further fractionation with methylcyanide and isopropanol gave three fractions: 1) a crude mixture of phytosterols, 2) resin, and 3) undigested organic matter. The crude mixture of phytosterols after purification on neutral alumina followed by GLC analysis resulted in the separation of 68.7% of -sitosterol, 18.4% of stigmasterol and 12.9% of campesterol and brassicasterol together. Phytosterols were extracted more easily from fermented than from unfermented samples, because of biodegration of lipophilic compounds by the methanogenic bacteria.Acknowledgments. S.K.S. and J.C.S. are thankful to the authorities of the U.G.C., New Delhi, for the award of Teacher-fellowships. R.C.S., R.A.K.S. and D.K.M. are thankful to the authorities of the C.S.I.R., New Delhi, for providing R.A., S.R.F., and J.R.F., respectively. Reprint request to S.N.M.     

The attenuation of prostaglandin-induced luteolysis in decidual tissue-bearing pseudopregnant rats

Summary Decidual tissue (DT)-bearing pseudopregnant (PSP) rats, in contrast to hysterectomized PSP rats, were resistant to a luteolytic regimen of either PGF₂ or PGE₂ when examined for the duration of PSP diestrus. The PG treatments, however, caused a marked fall in the serum progesterone levels on days 10 and 12, although the levels in DT-bearing rats on day 10 were significantly higher than in the hysterectomized rats.Acknowledgment. The authors wish to thank Helen Wilk and Edward Butler for their help in animal care; Ellen O'Laughlin-Phillips for the progesterone assays; Rosa Garnett, Jo Fletcher, and Aileen Svaty for editorial and secretarial help; Dr John E. Pike of the Upjohn Company for the gifts of PGE₂ and PGF₂; and Dr G. N. Niswender for the gift of antiserum GDN-337 used in the RIA for progesterone.Aided in part by NIH Training Grant HD-00024, Ford Foundation Grant 670-0135A, and NIH Program Project HD-07640.     

Anti-muscarinic activity of a family of C11N5 compounds isolated fromAgelas sponges

In a search for potential target sites for C₁₁N₅ compounds obtained from marine sponges of the genusAgelas we evaluated their interaction with muscarinic acetylcholine receptors from rat brain membranes. In competition experiments with³H-QNB these compounds displayed the following rank order of potency: sceptrin>oroidindibromosceptrinclathrodin. Sceptrin (50 M) was shown to be a competitive inhibitor of³H-QNB binding as revealed by Scatchard analysis. The results demonstrate the ability of these compounds to interact with multiple target molecules in the micromolar range.     

Hemmung der experimentellen,testosteron-bedingten Epidermisproliferation an der weiblichen Ratte durch ein synthetisches Phenanthrenderivat mit antiandrogener Wirkung

Summary In the female rat, testosterone propionate-induced proliferation of the epidermis (acanthosis and mitotic activation) is inhibited in proportion to dosage by a phenanthrene derivative with antiandrogenic action.     

Heavy water intake in tissues. II. H2O−D2O exchange in the myelinated nerve of the frog

Summary The kinetic parameters of D₂O intake in the frog sciatic nerve are obtained by means of infrared spectroscopy. 3 aqueous compartments: a non-exchangeable one (29%) and 2 compartments of quasi-free exchangeable water: 50% intracellular and 21% extracellular, are revealed.The first communication in this series is V. Vasilescu, D.-G. Mrgineanu and Eva Katona, Naturwiss.62, 187 (1975).For such derivations reference is made to the above mentioned mathematical model or to related works on compartmental analysis, reviewed by A. Rescigno and J. S. Beck, in: Foundations of Mathematical Biology, vol. II. Ed. R. Rosen, Academic Press, New York 1972.     

The identification of ecdysterone (20-hydroxyecdysone) in 3 species of molluscs (Gastropoda: Pulmonata)

Summary Ecdysterone has been identified in the schistosomiasis vectorBiomphalaria glabrata Say and inHelix aspersa Müller as well as inLymmaea stagnalis L. by chromatography, bioassay, radioimmunoassay, derivatization and by mass spectroscopy. Analysis of the food, faeces and hepatopancreas suggest that the sterol is derived from the diet. The probable function of ecdysterone in relation to calcification of the shell is discussed in this paper.We wish to thank Professor P. Karlson and Dr J. Koolman (Phillips University, Marburg-Lahn, FRG) for providing facilities for RIA. The initial study was begun at the University of Sussex and continued at the Courtauld Institute of Biochemistry, Middlesex Hospital Medical School, London, while D.L.W. was supported by the Ministry of Overseas Development. London and the Medical Research Council of Great Britain (grant G974/525/T). We thank Mr P. Mellon for the mass spectrum and Dr G.B. Russell for his interest in the earlier part of this study. We also thank Professor E. Scherer and Dr Q. Uwe (University of Essen, FRG) for their help in the irradiation ofL. stagnalis.Gauss Str. 82, D-4630 Bochum 5 (Federal Republic of Germany). Research results usingL. stagnalis were submitted for the Philips Europa Wettbewerb für junge Forscher und Erfinder (1979) in a thesis entitled Zur Frage der Existenz von Steroiden und Gonadoliberin bei der Grossen Schlammschnecke (Lymmaea stagnalis L.): ein Beitrag zur Evolution der Insekten- und Wirbeltierhormone.     

An investigation of the interaction between isoniazid and the contraceptive steroid norethindrone in vivo

Summary Isonicotinic acid hydrazide (isoniazid) was shown to react readily with 17-ethinyl-17-hydroxyestr-4-en-3-one (norethindrone) to form the isonicotinyl hydrazone of the steroid under conditions likely to exist in the stomach. The hydrazone was detected in guinea-pig, but not rat, plasma following its oral administration. Rat liver tissue metabolized the compound more rapidly than guinea-pig liver in vitro which probably accounts for the failure to detect the hydrazone in rat plasma.     

Amylin given by central or peripheral routes decreases gastric emptying and intestinal transit in the rat

The effect of rat amylin on gastric emptying and intestinal transit in the rat was examined. Amylin administered intracerebroventricularly (1, 2, 2.5 or 4 g/rat) produced the maximal decrease in gastric emptying and intestinal transit at the dose of 2.5 g/rat. Higher doses produced a lower effect. Peripheral administration (25, 50 or 100 g/kg) produced dose-dependent effects. Pre-treatment with neostigmine blocked the effect of amylin when it was centrally injected, while the effect of amylin given peripherally was partially reduced. Pre-treatment with domperidone decreased the inhibitory effect of peripherally injected amylin, but no effect was observed when the peptide was centrally injected.