Volume 14, Issue 3
September 1989, pages 203-328
pp 203-208 September 1989
Endogenous polyphosphate depletedAnabaena ARM310, solubilized extracellular tricalcium phosphate through increased phosphatase activity.
pp 209-219 September 1989
Actin like protein, extracted and purified fromVigna radiata (mung bean) seedling, has been found to give positive enzyme-linked immunosorbent assay with mouse monoclonal antiactin antibody.
In vivo studies show that cytochalasin B at sublethal dose inhibits the chromosomal movement at metaphase stage during germination. Fromin vitro studies it is found that the actin like protein isolated from mung bean seedling has a cytochalasin B binding property with a Kd value 1.2 × 10−5 M. From these two specific observations it appears probable that the biological function of mung bean actin like protein is to take part in cell division process directly or indirectly during the time of seedling development.
pp 221-231 September 1989
The purified biotin binding protein of pregnant rat serum was shown to be immunologically similar to rat serum albumin as assessed by a sensitive radioimmunoassay. In radioimmunoassay for rat biotin binding protein, the binding of [125I] rat biotin binding protein to anti-chicken egg yolk biotin binding protein antibodies was displaced by both rat serum (10–100 nl) and purified rat serum albumin (0.1–10 ng). Similarly, in radioimmunoassay for rat serum albumin the binding of [125I] rat serum albumin to either anti-rat serum albumin antibodies or anti-chicken egg yolk biotin binding protein antibodies was displaced by unlabelled rat biotin binding protein at comparable concentration range (0·5–10 ng). Significant fractions of radioiodinated rat biotin binding protein and rat serum albumin bound to antibodies to chicken egg yolk biotin binding protein. In immature rats, the circulating half-lives of rat biotin binding protein and rat serum albumin were determined to be 12 and 17 h respectively. The rat biotin binding protein and rat serum albumin were analysed by techniques that exploit their physicochemical properties. They displayed similar electrophoretic mobilities in alkaline as well as denaturing sodium dodecyl sulphate-polyacrylamide gels. However, in nonequilibrium pH gradient polyacrylamide gel electrophoresis, they resolved clearly. In two-dimensional tryptic peptide map analysis, the two proteins showed similarities as well as significant differences in the relative distribution patterns of their iodopeptides. These results showed that the primary structure of rat biotin binding protein and rat serum albumin were different in finer details despite the fact that they shared significant immunological cross-reactivity.
pp 233-241 September 1989
A single intraperitoneal injection of DL-methionine (500 mg/kg body wt.) to adult male Wistar rats was shown to significantly induce all the components of the hepatic microsomal mixed function oxidase system such as NADPH cytochrome C reductase activity, cytochromes P-450 and b5, as well as activities of drug metabolizing enzymes such as aminopyrine demethylase and uridine 5′ -diphosphate-glucuronosyltransferase. Combined administration of nicotinamide (250 mg/kg body wt.) and DL-methionine (500 mg/kg body wt.) was shown to bring about an additional increase (25-30%) in the activities of these enzymes as compared to their induction on independent administration of the two endobiotics. In rats bearing Yoshida sarcoma (ascites) tumour as well as in normal rats injected with serum from tumour bearing animals, the decreased activities of hepatic mixed function oxidases could be restored to their normal levels by administration of DL-methionine (500 mg/kg body wt.) to these rats. Whereas actinomycin D (1 mg/kg body wt.) had no effect on the increased incorporation of [14C] labelled leucine into microsomal proteins following administration of nicotinamide, the enhanced incorporation of the label following DL-methionine administration was completely inhibited by the same dose of actinomycin D. Administration of cycloheximide (0·5 mg/kg body wt.) to rats could completely inhibit the increased incorporation of [14C] leucine into hepatic microsomal proteins following independent administration of nicotinamide and DL-methionine. Similar inhibitory pattern with actinomycin D and cycloheximide was also demonstrated in case of induction of NADPH cytochromeC reductase activity by both these endobiotics.
pp 243-247 September 1989
The effect of doxorubicin on glucose metabolism was studied in rats with or without the supplementation of α-tocopherol. Rats were treated with doxorubicin, 2 mg/kg body wt. (intravenously), twice a week, for 6 weeks. α-Tocopherol (400 mg/kg body wt.) was co-administered orally for 2 months. Glycolysis was found to be increased with a significant decrease in the activities of tricarboxylic acid cycle enzymes. A significant increase in liver glycogen was noted in doxorubicin treated rats. Activities of glycogen Phosphorylase, glucose-6-phosphate dehydrogenase and glucose-6-phosphatase were found to be decreased.
α-Tocopherol co-administration was found to reduce the alterations in the above mentioned enzyme activities. The results are discussed with reference to the drug metabolism, lipid peroxidation and the antioxidant nature of α-tocopherol.
pp 249-253 September 1989
A sensitive staining method was developed to localise the activity of myo-inositol-1-phosphatase on Polyacrylamide gels after electrophoresis. The method can also be used for non-specific phosphatases as well as for those specific phosphatases acting upon inositol polyphosphates which are prime cellular second messengers. One or two nmol of phosphate is sufficient and less than 3 µg of purified protein will facilitate the localisation of phosphatase. If more phosphatases are present in the enzyme preparation, a combination of inhibitors can be used to suppress the activities of unwanted phosphatases and the use of specific substrates will facilitate the localisation of enzyme of interest.
pp 255-260 September 1989
A feasibility study of neural transplantation in adult rhesus monkey was undertaken. Fresh and preserved neocortex containing multiplying and maturing neurons obtained from 55–70 gestation days were transplanted into the striatum, cerebellum and cerebral cortex of adult monkeys. Tissues were preserved for 4 days either at subzero temperature in the freezer compartment of the ordinary refrigerator in Ringer lactate or incubated in culture medium. While 2 monkeys out of 5 injected with preserved tissue had successful transplants after 4 months, all the 10 monkeys injected with fresh tissue had no transplants. The size of the two surviving transplants was small. The neurons in the transplants were mainly in clusters. Many of the cells were immature and some showed early degenerative changes. Neuronal processes were restricted to the transplants and thus showed lack of morphological integration with the host tissue. Further studies are in progress to define the nature of the embryonic tissue of primate which can grow and survive and also the role of neural grafts in functional recovery following experimental lesions of the brain regions.
pp 261-268 September 1989
Purification of cathepsin B from buffalo-spleen, a hitherto unstudied system has been achieved by a simple procedure developed by incorporating suitable modifications in the existing methods for isolation of the enzyme from other sources. The purified enzyme has a molecular weight of 25 KDa and its Stokes radius was found to be 2·24 nm. Effects of several reducing agents, urea and thiol-protease inhibitors such as leupeptin and antipain, have been studied and the data unequivocally support the contention that the buffalo-enzyme is similar to cathepsin B from other tissues with respect to these properties.
pp 269-277 September 1989
Highly pure lysosomes were isolated from buffalo(Bubalus bubalis) kidney cortex by a procedure involving differential and isopycnic Percoll density gradient centrifugations. Arylsulphatase, N-acetyl-Β-glucosamindase and cathepsin D in the lysosomal preparation were 26–45-fold enriched over the homogenate. The purified lysosomes contained less than 0·06% of mitochondrial, microsomal and peroxisomal marker enzymes. In the electron micrographs the particles appeared as large dense granules of size 0·3-1·9 µm with no apparent structural features belonging to mitochondria or microsomes. The isolation procedure was also found to be suitable to obtain highly pure lysosome particles from renal cortex of other sources such as rat, lamb and beef. No ultracentrifugation steps were involved in the procedure
pp 279-287 September 1989
A DNA hybridization assay was developed using a cloned hepatitis B viral genome to detect the presence of infectious virions in human serum. The merit of this assay was to put in evidence virus particles in 7 out of 133 sera that were negative for surface antigen (HBsAg) using routine serological methods. The usefulness of this assay was confirmed by actual visualization of the virus under electron microscope. Some serum samples although positive for surface antigen, did not give a hybridization signal by dot blot assay and might indicate cases of acute hepatitis
pp 289-299 September 1989
An aliphatic segmented polyurethane with soft to hard segment ratio 3 was synthesised using hexamethylene diisocyanate, polypropylene glycol 400 and 1,4-butane diol.A stainless steel cage implant system has been used to study thein vivo biocompatibility of this polyurethane. United States Pharmacopoeia negative control polyethylene was used for the comparison. Three cages, one with polyurethane another with United States Pharmacopoeia polyethylene and the third control empty cage were implanted subcutaneously in the dorsal aspect of rabbits. The inflammatory exudate surrounding the material was aspirated from the cages on 4, 7, 14 and 21 days after implantation. The total protein content in the exudate aspirated from all the 3 cages was significantly higher at 7 days than in the reported normal rabbit serum of New Zealand white rabbit but equal to that of our rabbit colony. The albumin concentration was lower in the initial period but increased at 21 days post implantation period in all the cages. Concentration of α1, α2 and γ-globulin also decreased in all cages at 21 days. Neutrophils were predominant in all the exudates aspirated from polyurethane, polyethylene and empty control cages during whole implantation period. This is attributed to the profound effect of the cages on the surrounding vasculature. Macrophage was found to be seen during acute phase of inflammation due to the migration of macrophage along with neutrophil towards the inflammatory lesion. The percentage of neutrophils showed a faster decline in the cage containing polyethylene at 21 days. The extra cellular alkaline phosphatase activity, though higher in exudate from cages containing polyurethane at 14 days post implantation, was same in all 3 cages at 21 days. Leucine amino peptidase activity was found to be decreased at 21 days of post implantation time though the empty control cage exhibited an increase at 14 days post implantation. The inflammatory response at 21 days was similar in polyurethane and the control polyethylene
pp 301-309 September 1989
A pectin present in the juice of the inflorescence stalk of plantain(Musa sapientum) has been isolated. The material contained 32.4% hexoses and 52.5% uronic acid. On administration to rats fed both cholesterol free and cholesterol diet, this material showed significant lowering of cholesterol and triglycerides in the serum, liver and aorta. There was decreased cholesterogenesis in the liver as was evident from decreased activity of hydroxymethylglutaryl coenzyme A reductase and decreased incorporation of labelled acetate into hepatic cholesterol. Hepatic bile acids showed significant increase and there was increased fecal excretion of neutral sterols and bile acids. Release of lipoproteins into the circulation was lower. The material also caused increase in the activity of lipoprotein lipase in the heart and adipose tissue and also of plasma lecithin: cholesterol acyl transferase
pp 311-317 September 1989
Improved solid phase synthesis of luteinizing hormone releasing hormone analogues using 9-fluorenylmethyloxycarbonyl amino acid active esters and catalytic transfer hydrogenation with minimal side-chain protection and their biological activities
Using mainly 9-fluorenylmethyloxycarbonyl amino acid 2, 4, 5-trichlorophenyl esters in the presence of 1-hydroxybenzotriazole and the solid supportp-alkoxybenzyl alcohol resin, synthesis of luteinizing hormone releasing hormone analogues was carried out with minimal side-chain protection. Catalytic transfer hydrogenation was employed for removal of NO2 and Z-groups from Arg and < Glu respectively avoiding the use of HF and this led to good yields. An aromatic, hydrophilic amino acid, D-(p-hydroxyphenyl) glycine was incorporated into luteinizing hormone releasing hormone molecule along with other modifications. The agonistic as well as antagonistic activities of all the peptides have been studied
pp 319-328 September 1989
The quantum mechanical perturbation method has been utilized to study the biological activity of 8-azapurine (8-azaguanosine, 8-azaadenosine and 8-aza-2,6-diamino-purine) nucleoside antibiotics. The in-plane (hydrogen bonding) and stacking energy of 8-azapurine bases have been evaluated with nucleic acid bases and base pairs in all possible orientations. The energy values and the sites of association of analogous bases, obtained by optimization of energy values as well as the sites of association of nucleic acid bases during the transcription process have been compared. The model developed earlier for the incorporation of nucleoside analogues has been used to find out the inhibitory effects of the drug on nucleic acid and protein synthesis. It has been observed that the activity of 8-azapurines are of the following order
8-azaguanine > 8-aza-2,6-diaminopurine > 8-azaadenine
and these analogues show preference for binding near a guanine or cytosine in the chain. The results are in agreement with the experimental observations