Volume 1, Issue 2
June 1979, pages 109-253
pp 109-124 June 1979
Diamine oxidase (EC 18.104.22.168) was purified from 5-day-old etiolated seedlings ofLathyrus sativus by MnCl2 treatment, (NH4)2SO4 and acetone fractionations, DEAE-Sephadex chromatography followed by gel filtration on Sephadex G-200. A single step purification of the enzyme was achieved by using an immunoaffinity column, wherein rabbit antibodies to the homogeneous diamine oxidase were coupled to CNBr-activated Sepharose. The enzyme thus obtained was homogeneous by electrophoretic, immunological and ultracentrifugal criteria. It had anMr of 148,000 (6.46S) and was a dimer with similar sub-units (Mr 75,000). Amino acid analysis showed the absence of cysteine residues although it contained five disulphide bonds. The enzyme had copper (2.7 g atom/mol enzyme) but was not a glycoprotein. No absorption maximum in the visible region was detectable. Ethylenediamine 1,3-diaminopropane and histamine were potent competitive inhibitors for the substrate putrescine. The addition of monospecific antibodies to the enzyme increased the Km for benzyl amine without any change in the Vmax Diamine oxidase from pea seedling, partially purified, exhibited complete crossreactivity with the antibodies to theL. sativus enzyme.
pp 125-132 June 1979
The interaction of allylisothiocyanate with bovine serum albumin was monitored by fluorescence titration. The interaction was weak with an apparent association constant of 2 × 102. The interaction was unaffected in the pH range of 5.0 to 8.3 and by NaCl. However, the addition of dioxane upto 4% increased the value of the association constant. N-Methyl bovine serum albumin and bovine serum albumin with sulphydryl groups blocked had the same affinity for allylisothiocyanate suggesting that amino and sulphydryl groups may not be involved in the interaction. Polyacrylamide gel electrophoresis and estimation of available lysine suggested that there were perhaps two types of groups involved in the interaction of allylisothiocyanate with bovine serum albumin.
pp 135-141 June 1979
Some aspects of uterine RNA synthesis including [3H]-uridine incorpo ration into RNA, activities of RNA polymerases and ribonucleases were studied. It was observed that both normal and pregnant animals, kept on protein-free diet for 15 and 20 days, showed a significant increase inin vivo uptake of [3H]-uridine into total RNA. Activities of RNA polymerase I and polymerase III increased two-fold in animals kept on a protein-free diet; however, RNA polymerase II activity was unaffected by protein restriction. In animals kept on protein-free diet where pregnancy was maintained by exogenous estrogen and progesterone, specific activity of nuclear RNA was further increased and the activities of RNA polymerases I, II and III markedly increased. Levels of RNase were also increased significantly during protein deficiency, thus showing a rapid turn-over of uterine RNA. These observations indicate that during protein restriction, uterine RNA synthesis is regulated at transcriptional level by a selective stimulation of RNA polymerase and RNase also plays an important role.
pp 143-149 June 1979
The inhibition of growth of a wild strain ofNeurospora crassa by Cu2+ is counteracted by histidine, histidine methyl ester, histidinol and Mn2+. In the presence of Cu2+, the total free amino acid content decreased by 30%. The decreased free amino acid pools of arginine, histidine and tyrosine were restored on the addition of Mn2+. Histidinol phosphate phosphatase showed a decrease in activity in the presence of Cu2+. This inhibition was reversed on the addition of excess Mn2+. The data suggest that copper toxicity in the mould is due to suppression of histidine biosynthesis.
pp 151-157 June 1979
Addition of sonicated dispersions of cholesterol to peptone-salt-vitamin medium resulted in the metabolism of the sterol byHartmanella culbertsoni. Trophozoite multiplication was stimulated at 1–5 mg/litre, but retarded at 10–20 mg/litre. When cholesterol was added to the medium, incorporation of [1,2-14C] -acetate into neutral lipid, phospholipid, non-saponifiable and cholesterol fractions of the amoebae was significantly reduced. Cholesterol ester was detected in the medium but phospholipids were not released. Addition of cholesterol stimulated the activity of lysosomal acid phosphatase, acid deoxyribonuclease and cathepsin B but did not affect 5′-nucleotidase, adenosine triphosphatase, alkaline phosphatase, glucose-6-phosphatase, succinate dehydrogenase and cytochrome C oxidase.
pp 159-168 June 1979
An enzyme catalysing the hydrolysis ofa-tocopheryl acetate was characterised in chicken liver. The enzyme was localised in the microsomes, had an optimum pH 8.6 and aKm value of 0.5 mM. The enzyme did not hydrolyse retinyl acetate, cholesteryl acetate and ethyl acetate, thus indicating a high degree of specificity.a-Tocopheryl acetate hydrolase required bile salts as a specific co-factor. The results suggested a role for this enzyme in the absorption of vitamin E.
pp 169-177 June 1979
The effects of ethidium bromide, an intercalating dye and berenil, a nonintercalating dye on the biological activities ofEscherichia coli ribosomes have been studied. Ethidium bromide treatment drastically reduced both enzymatic and nonenzymatic initiation complex formation, enzymatic as well as nonenzymatic binding of phenylalanyl tRNA, peptidyl transferase, GTPase as well as the overall protein synthesising activity as measured by the poly U-dependent polymerization of phenylalanine. On berenil treatment, however, only enzymatic formation of the initiation complex is marginally reduced. Other reactions are not markedly affected except the enzymatic phenylalanyl tRNA binding which is slightly decreased only at high Mg2+ concentration; the treated ribosome has lowered polymerizing activity at sub-optimal Mg2+ concentration (10 mM). Although it has already been shown in this laboratory that treatment with either dye leads to the unfolding of the structure of the ribosome, the present studies indicate that berenil treatment does not alter the structure of the ribosome drastically in contrast to ethidium bromide treatment.
pp 179-183 June 1979
A comparative study of the distribution of a simple esterase and acetylcholinesterase in the cerebellar cortex of mouse and bat has been made. The Purkinje layer is intensely positive for simple esterase in both species. The granular and molecular layers showed mild to moderate activity in mouse and intense activity in bat. Acetylcholinesterase in cerebellar layers of bat is more intense than in mouse. In bat cerebellum, acetylcholinesterase is observed in the dendrites of Purkinje cells, but not in their cell bodies. Acetylcholinesterase was not found in Purkinje cells of mouse.
pp 185-205 June 1979
The present study was carried out to determine the detailed histological and cytological features of the excurrent ducts of the male reproductive system in the rhesus monkey. The excurrent ducts show a regional difference in their histological features. The use of some of these features as histological markers and their possible functional significance are discussed.
The epithelial cells in the different components of the excurrent duct system possess cytological features which suggest their involvement in absorption and the secretion of different products into the lumen.
pp 207-213 June 1979
The fat body appears to contribute cholesterol for testicular steroidogenesis. It also appears to provide prostaglandins and cyclic AMP for testicular steroidogenesis since fatectomy impairs this process which is corrected by the addition of prostaglandins and cyclic AMP. Of the two, prostaglandins have a more important role in spermatogenesis and cyclic AMP functions in steroidogenesis These functions of the fat body suggests that it constitutes a link in the hypothalamohypophysial-gonadal axis.
pp 215-222 June 1979
A number of equations for the various population control policies are worked out for a desired reduction in the rate of growth. At the ages of 25 and 30 respectively, 61 and 97% of contraceptive users are necessary to reduce the present rate of growth of 0.026 to 0.010. While at the age of 25 about 69 and 76% contraceptive users are required for the same reduction in the rate of growth, assuming that 25 and 50% would discontinue the use of contraceptives at the age of 35. The birth and death rates in the study area (Varanasi Rural) have remained almost constant for several years, justifying the assumption of a stable population. This study emphasises the need for the use of contraceptive devices at two or more age levels.
pp 223-234 June 1979
Metabolism of isonicotinic acid and isoniazid bySarcina sp. led to the formation of two metabolites which were characterised as 2-hydroxyisonicotinic acid and citrazinic acid. The blue pigment formed during fermentation was shown to be derived from the auto-oxidation of citrazinic acid. 2-Oxo-glutarate accumulated as the major keto acid when isonicotinic acid or isonicotinic acid hydrazide metabolism was inhibited by 1 mM sodium arsenite. Isonicotinic acid, 2-hydroxy-isonicotinic acid and 2-oxo-glutarate were oxidised by isonicotinic acid hydrazide or isonicotinic acid-grown cells; citrazinic acid was, however, not oxidised. Isoniazid hydrazine hydrolase, isonicotinic acid and 2-hydroxyisonicotinic acid hydroxylases were detected in the cell-free extract ofSarcina sp. grown on isonicotinic acid hydrazide or isonicotinic acid.
pp 235-239 June 1979
An actinomycete was isolated during a soil screening programme to obtain L-3,4-dihydroxyphenylalanine producers. A mutant of this organism was isolated by chemical mutagenesis and it accumulated 1 g/litre L-dihydroxyphenylalanine when grown on L-tyrosine. Resting cells converted 30% of tyrosine in the reaction mixture. The use of resting cells for dihydroxyphenylalanine production is advantageous as it eliminates interfering substances which accumulate during fermentation.
pp 241-253 June 1979
Samples of freeze dried green field bean (Dolichos lablab) and dry mature bean, were subjected to the following processing methods—heat processing, extraction with 80% ethanol, hexane or dilute acid, protein isolation; and these samples were evaluated for growth promoting value and toxicity. Extraction with 80% ethanol or with dilute acid increased survival period of the animals; but these did not promote growth. Heat processing was essential to destroy antinutritional factors and promote growth. Extraction of the beans with 80% ethanol did not however alter the trypsin inhibitor or haemagglutinin activities. The protein isolate and acid-extracted residue which had low trypsin inhibitor and haemagglutinin activities, did not also promote growth. Thus the trypsin inhibtor and haemagglutinin activities did not completely account for the toxicity to albino rats. However, heat processing of ethanol extracted bean flour indicated that the beneficial effect of ethanol extraction was not apparent, once the samples were heat processed. Dry mature bean dhal was more toxic than the whole bean either dry or green. Supplementation of heat processed field bean with methionine and tryptophan promoted good growth of albino rats and significantly increased the protein efficiency ratio.