• Volume 15, Issue 1

      March 1990,   pages  1-58

    • Affinity chromatography of estrogen- and progesterone-binding proteins of human uterus

      M Thapar P Sujata G L Kumari T G Shrivastava S K Sachdeva

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      The purification of estrogen- and progesterone-binding proteins of human uterus by employing affinity resins coupled with steroid-bovine serum albumin conjugates, led to the isolation of preparations with estrogen- and progesterone-binding sites havingKd values in the range of 0.96 to 1.20 × 10-9 M. These were different from theKd values of 10-10 M and 10-8 M obtained for two types of binding sites present in the crude cytosolic and nuclear fractions. The purified proteins sedimented on sucrose gradient withS values in the range of 3.6–4.4.

      The cytosolic and nuclear estrogen- and progesterone-binding proteins, thus purified, showed differences in specificity of binding to the hormone. While the cytoplasmic proteins were more specific in their binding to estradiol or progesterone, the nuclear proteins bound Cortisol with equal or moderate affnity. These results demonstrate the presence of distinct physiological forms of estrogen- and progesterone-binding proteins in the cytoplasm and nucleus, thus pointing to the importance of both these compartments in hormone action.

    • Cholecystokinin antagonist, proglumide, stimulates growth hormone release in the rat

      E Vijayan S M McCann

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      Previous studies have revealed a stimulatory action of cholecystokinin on growth hormone release in the rat. To evaluate the physiologic significance of these effects we employed the cholecystokinin antagonist, proglumide and injected it intravenously and intraventricularly (third cerebral ventricle, 3V) to determine its actions on growth hormone. The experiments were performed in conscious, freely moving rats with indwelling cannulae in the 3V and/or external jugular vein. Intraventricular injection of 2 or 10 □g of proglumide significantly elevated plasma growth hormone concentrations in intact and castrated male rats and in ovariectomized females. Intravenous injections of 10 or 100 □g of proglumide were also effective in elevating growth hormone in a dose-related manner. Surprisingly, the response to the lower dose given intraventricularly was somewhat greater than that of the higher dose. We speculate that these stimulatory effects of proglumide given intraventricularly are due to the agonist action of proglumide at these doses since action of cholecystokinin itself is to increase plasma growth hormone following its intraventricular injection. The studies therefore do not establish a physiologically significant growth hormone-releasing action of brain cholecystokinin but provide more evidence that activation of cholecystokinin receptors in the brain can induce a stimulation of growth hormone release either by activation of the release of growth hormone-releasing hormone or by inhibition of the release of somatostatin or by a combination of these two actions.

    • Non-enzymatic glycosylation induced changesin vitro in some molecular parameters of collagen

      A N Rathi G Chandrakasan

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      Non-enzymatic glycosylation of rat tail tendon collagen was examined by incubation with D-glucosein vitro. The changes in molecular parameters such as viscosity, thermal stability, electrophoretic mobility and solubility were determined on nonenzymatically glycosylated collagenin vitro. Tendons incubated with 8 and 24 mg glucose/ml showed an increase in dissolution temperature and a l.6-3-fold increase in thermal isometric tension respectively when compared to tendons incubated in the absence of glucose, indicating the formation of new intermolecular bonds. This conclusion was further supported by the decreased solubility of glycosylated collagen in 0.5 N acetic acid and the change in sub-unit composition as measured from the sodium dodecyl sulphate Polyacrylamide gel electrophoresis pattern. Glycosylated collagen gave a characteristic absorption spectra λmax 248 nm) as distinct from that of control (λmax 242 nm). Denaturation temperature of glycosylated collagen, as determined from temperature dependent viscosity measurements, was reduced. These studies indicate that glycosylation affects the molecular interactions as well as the crosslinking of collagen.

    • Effect of doxorubicin on the intestinal membranes in rats—Influence of α-tocopherol administration

      A Geetha R Sankar Thankamani Marar C S Shyamala Devi

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      The effect of α-tocopherol on doxorubicin induced changes in intestinal brush border and basolateral membranes were studied in rats. Rats were treated with doxorubicin (2.5 mg/kg body wt.), intravenously, weekly for 8 weeks. α-Tocopherol (400 mg/kg body wt.) was given orally, daily for 2 months. Intestinal basolateral membrane bound ATPases and brush border membrane bound alkaline phosphatase activities were found to be decreased significantly in doxorubicin treated rats. The lipid peroxide level was found to be elevated with a significant depletion in membrane sulphydryl groups. In α-tocopherol coadministered animals, the enzyme activities were found to be restored with concomitant reduction in lipid peroxide levels and an increase in the membrane sulphydryl groups. The membrane cholesterol and phospholipid levels which were altered in doxorubicin treated animals were found to be maintained significantly. The results are discussed with reference to the effect of α-tocopherol on lipid peroxidation and membrane sulphydryl groups.

    • Analysis, characterization and diagnostic use of circulating filarial antigen in bancroftian filariasis

      K A Parkhe M V R Reddy K Cheirmaraj P Ramaprasad B C Harinath

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      Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of circulating filarial antigen fraction-2 isolated from plasma of microfilaraemic patients withWuchereria bancrofti infection has shown 21 bands with molecular weights ranging from 12 to 120 kDa. The gel (12 cm) was sliced at an interval of one cm and the eluates of all the gel slicesviz., CFA2-1 to CFA2-12 showed the presence of filarial antigen by sandwich enzyme-linked immunosorbent assay. The low molecular weight circulating filarial antigen fractions were found to share a common epitope withWuchereria bancrofti microfilariae excretory-secretory antigen and urinary filarial antigen. The 3 antigen fractions CFA2-1, CFA2-9 and CFA2-12 showed higher sensitivity in detecting filarial immunoglobulin M antibodies than immunoglobulin G antibodies. However CFA2-9 fraction was found useful in serological differentiation of microfilaraemics from those with disease manifestations when filarial immunoglobulin G antibodies were detected. The antigenic epitope of CFA2-1 appears to be a carbohydrate, whereas CFA2-9 appears to be protein in nature.

    • 5-Methylcytosine content and methylation status in six millet DNAs

      Lalitha S Kumar R R Hendre P K Ranjekar

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      High performance liquid chromatographic analysis of the total nuclear DNAs of 6 millets plant species indicates that the 5-methylcytosine content ranges from 3% in barn yard millet to 9.6% in great millet while the fraction of cytosines methylated varies between 14% in little millet to 31 % in pearl millet. Digestion of millet DNAs with MspI/HpaII suggests that CpG methylation is more in great millet DNA while CpC methylation is more in the other 5 millet DNAs. Digestion of millet DNAs with MboI, Sau3AI andDpnI indicates that some of the5’ GATC3’ sequences are methylated at adenine and/or cytosine residues except in little millet where adenine methylation of the5’GATC3’ sequences is insignificant and there is a predominance of cytosine methylation in these sequences.

    • Predicted secondary structure of maltodextrin Phosphorylase fromEscherichia coli as deduced using Chou-Fasman model

      Anil Kumar

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      Secondary structure of maltodextrin Phosphorylase fromEscherichia coli has been predicted using Chou-Fasman model. The enzyme protein contains 28% α-helix, 27% β-pleated sheets and 20% reverse β-turns. The secondary structure predicted 4 regions showing Rossman-fold super secondary structure. Two regions, one from residue 268–361 and the another from residue 606–684, having 4 consecutive strands of parallel β-pleated sheets and 3 joining α-helix, are predicted. Two regions, one from residue 379–434 and the another from residue 496–573, having 3 consecutive strands of parallel β-pleated sheets and two joining α-helix, are predicted.

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