Assessment of the genotoxic potential of riboflavin and lumiflavin. B. Effect of light.

On exposure to visible light, riboflavin and lumiflavin produced reactive oxygen species such as singlet oxygen and superoxide radicals. The reaction was found to be time- and concentration-dependent. Both riboflavin and lumiflavin, upon illumination, showed mutagenic response in the umu test as well as in the Ames/Salmonella assay with Salmonella typhimurium TA102. The mutagenic response was partially abolished by superoxide dismutase while sodium azide did not have any effect. No mutagenicity was observed if the compounds were not illuminated. The results suggested the involvement of superoxide radicals in light-induced mutagenicity of riboflavin as well as lumiflavin.     

Reassessment of the prevalence of heat-stable enterotoxin (NAG-ST) among environmental Vibrio cholerae non-O1 strains isolated from Calcutta, India, by using a NAG-ST DNA probe.

A collection of 521 environmental isolates of Vibrio cholerae which were previously examined by the suckling mouse assay and found to be negative for the heat-stable enterotoxin NAG-ST were reassessed by a recently developed DNA probe for NAG-ST. A total of 12 (2.3%) of the isolates hybridized with the NAG-ST probe. By using a cholera toxin (CT) DNA probe, the CT gene was detected in six of the strains in the collection, although none of the isolates of V. cholerae non-O1 hybridized with both of the toxin probes. All of the NAG-ST and CT probe-positive strains were hemolysin positive. Thirty-fold-concentrated supernatants of the three representative NAG-ST DNA probe-positive V. cholerae non-O1 strains gave positive fluid accumulation ratios in the suckling mouse assay even after heating (100 degrees C for 5 min) and also inhibited the binding of a NAG-ST monoclonal antibody to the bound NAG-ST in a competitive enzyme-linked immunosorbent assay (ELISA). Likewise, all six CT probe-positive V. cholerae non-O1 strains produced in vitro CT when examined by the CT bead ELISA. HindIII digest patterns of chromosomal DNA from the representative NAG-ST gene-positive strains were visually indistinguishable. Between the groups of NAG-ST probe-positive strains examined, there was a variation in the hybridizable fragments, with one group of strains exhibiting a hybridizable fragment similar to that of the NRT 36 reference strain; a smaller HindIII fragment hybridized with the NAG-ST probe in the other group of strains.(ABSTRACT TRUNCATED AT 250 WORDS)     

Copper influenced changes of lipid metabolism in the tissues of freshwater teleost Labeo rohita (Hamilton).

Modulations in the lipid metabolism in the gill, liver, muscle and brain of freshwater teleost Labeo rohita exposed to 1, 2 and 3 days to lethal (1.2 mg/l) and 1, 15 and 30 days to sublethal (0.24 mg/l) concentrations of copper were studied. The total lipids decreased and there was an increase in the free fatty acids, glycerol and lipase activity in the organs studied at lethal concentration of copper. The degree in these shifts increased over time of exposure (1 less than 2 less than 3 days). In sublethal concentration, the levels of total lipids, free fatty acids, glycerol and lipase activity increased in all the four organs and the shifts followed two different trends during the exposure periods, 1 less than 15 less than 30 days in total lipids and 1 greater than 15 greater than 30 days in the other parameters. Among the organs, in both concentration media the changes in the lipid metabolism were in the order liver greater than gill greater than muscle greater than brain.     

Isolation and Identification of Vesicular-Arbuscular Mycorrhiza-Stimulatory Compounds from Clover (Trifolium repens) Roots

Two isoflavonoids isolated from clover roots grown under phosphate stress were characterized as formononetin (7-hydroxy,4′-methoxy isoflavone) and biochanin A (5,7-dihydroxy,4′-methoxy isoflavone). At 5 ppm, these compounds stimulated hyphal growth in vitro and root colonization of an undescribed vesicular-arbuscular mycorrhiza, a Glomus sp. (INVAM-112). The permethylated products of the two compounds were inactive. These findings suggest that the isoflavonoids studied may act as signal molecules in vesicular-arbuscular mycorrhiza symbiosis.     

Determination of cobalt in urine by gas chromatography—mass spectrometry employing nickel as an internal standard

A gas chromatographic—mass spectrometric method for the determination of cobalt in biological materials employing stable enriched ⁶²Ni as an internal standard and using lithium bis(trifluoroethyl)dithiocarbamate as a chelating agent is described. The method involves the addition of a known amount (1 μg) of ⁶²Ni to the sample, the formation of the chelate and the determination by selected-ion monitoring of the m/z ratio, which corresponds to . No appreciable memory effect was observed, and an acceptable dynamic range of 100 was found. There was good agreement between the cobalt concentration values determined by gas chromatography—mass spectrometry and electrothermal atomic absorption spectrometry. The present method has high sensitivity and can be used for the quantitation of cobalt at concentrations as low as 1 μg/l. The use of enriched ⁶²Ni circumvents the problem caused by endogenous nickel and simultaneously provides data on the nickel concentration in the biological sample without any additional experimental effort.     

An actidione resistantCandida tropicalis from custard apple juice

An actidione resistant yeast,Candida tropicalis, was isolated from fermenting custard apple juice. Though a slight inhibition of growth was observed on the first day with 5000 ppm of actidione, growth was equal to control after one week. Sorbic acid at 500 ppm and above inhibited the growth of this yeast while sodium benzoate and potassium metabisulphite were unable to suppress the growth even at 1000 ppm. Fermentation and assimilation of different carbon sources were delayed in the presence of 1000 ppm of actidione suggesting the disruption of protein synthesis by actidione. Contribution no. 985.     

Biochemical mechanism for the inhibition of phenylalanine ammonia-lyase induction in the absence of oxygen in potato tuber tissue

Induction of phenylalanine ammonia-lyase (PAL) in excised potato parenchyma tissue in the presence of light displayed a rigid requirement for oxygen. A     

X-Pro peptides: Solution and solid-state conformation of benzyloxycarbonyl-(Aib-Pro)2methyl ester,a type I β-turn

The synthesis of the tetrapeptide benzyloxycarbonyl(α-aminoisobutyryl-L -prolyl)₂-methyl ester (Z-(Aib-Pro)₂-OMe) and an analysis of its conformation in solution and the solid state are reported. Stepwise synthesis using dicyclohexylcarbodiimide leads to racemization at Pro(2). Evidence for the presence of diastereomeric tetrapeptides is obtained from 270-MHz¹H-nmr and 67.89-MHz ¹³C-nmr. The all-L tetrapeptide is obtained by fractional crystallization from ethyl acetate. The NH of Aib(3) is shown to be involved in an intramo-lecular hydrogen bond by variable-temperature ¹H-nmr and the solvent dependence of NH chemical shifts. The results are consistent with a β-turn conformation with Aib(1) and Pro(2) at the corners stabilized by a 4 → 1 hydrogen bond. The molecule crystallizes in the space group P2₁2₁2₁, with a = 8.839, b = 14.938, and c = 22.015 Å. The structure has been refined to an R value of 0.051. The peptide backbone is all-trans, and a 4 → 1 hydrogen bond, between the CO group of the urethane moiety and Aib(3) NH, is observed. Aib(1) and Pro(2) occupy the corner positions of a type I β-turn with ? = ?55.4°, Ψ = ?31.3° for Aib(1) and ? = ?71.6°, Ψ = ?38° for Pro(2). The tertiary amide unit linking Pro(2) and Aib(3) is significantly distorted from planarity (Δω = 14.3°).     

Aquatic oligochaeta from among the roots of eichhornia crassipes solms

The paper records the presence of sixteen species of aquatic oligochaetes, belonging to the Aeolosomatidae (1 species). Naididae (13 species) and Tubificidae (2 species), living in association with the floating roots of the water hyacinth, Eichhornia crassipes Solms., from a tank in Vizianagaram town in Andhra Pradesh, India. Of these, Aeolosoma hyalinum Bunke, 1967 and Dero obtusa D'Udekem, 1855 are new records for the Indian sub-continent.     

A quantitating reagent for methanethiolation of protein sulfhydryl groups

p-Nitrophenoxycarbonyl methyl disulfide has been synthesized for use as a quantitating agent for methanethiolation of protein sulfhydryl groups. This reagent reacts specifically and quantitatively with cysteine residues of proteins to yield an unsymmetrical disulfide containing a CH₃S group and concomitantly releases the chromophore, p-nitrophenol. Titration of the sulfhydryl groups of glyceraldehyde 3-phosphate dehydrogenase (EC 1.2.1.12) with this reagent has been studied. Incorporation of CH₃S as measured by the release of p-nitrophenol paralleled the loss of sulfhydryl group dependent activity of the enzyme. The enzyme was found inactive on modification of four of the eight sulfhydryl groups present in the enzyme. Stability of p-nitrophenoxycarbonyl methyl disulfide has also been studied in different buffer systems. The rate of decomposition of the p-nitrophenyl ester due to hydrolysis was found negligible below a pH of 8.0 compared to its rate of reaction with free sulfhydryl groups.