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Biomedical subjects

M Srivastava

Publications and source records attributed to M Srivastava.

At least 19 recordsLinked to original sources

Gonado-modulatory effects of high humidity is pineal dependent in Indian palm squirrel Funambulus pennanti (Wroughton).

F. pennanti presented a clear biphasic pineal mediated seasonal sexual cycle. This sexual cycle was essentially characterised by a very short period of sexual quiescence with an arrest of spermatogenesis during October-November. A small but clear decrease in sexual activity was also observed during March-April. This decrease in sexual activity, however, had no quantifiable effect on spermatogenesis. Sexual recrudescence was observed from December-January. The testes remained sexually active from January till September. Almost an inverse relationship was observed between pineal and testicular weight. Pinealectomy, however, prevented naturally induced gonadal regression during both the periods, i.e. September-November and February-April. Exposure of animals to high RH (80 +/- 4%) during sexually active phase induced a steep regression in testicular weight of sham-operated animals even in the presence of gonad stimulatory long photoperiod (16L:8D) and high temperature (40 degrees +/- 5 degrees C) while exposure of animals to moderate RH (65 +/- 5%) during sexual regression phase partially prevented testicular regression even in presence of inhibitory short photoperiod (11L:13D) and normal environmental temperature (30 degrees +/- 5 degrees C). Pinealectomized animals, neither exhibited testicular regression in February-March nor had involuted testes in September-October, thus, suggesting that the effect of humidity is mediated via the pineal gland.

Animals

Alternative splicing of human synexin mRNA in brain, cardiac, and skeletal muscle alters the unique N-terminal domain.

Several synexin (annexin VII) mRNAs have been identified by screening a human fibroblast cDNA library. One type of message contained an alternatively spliced cassette exon, predicting two isoforms of synexin differing in the N-terminal domain. Polymerase chain reaction analysis of synexin mRNA from various fetal and adult tissues, from human and monkey, revealed that the alternative splicing event is tissue-regulated; synexin mRNA containing the cassette exon is prevalent in brain, heart, and skeletal muscle. This is supported by Western blot analysis showing that muscle synexin (annexin VIIb) is larger than synexin from lung (annexin VIIa). The muscle and lung isoforms have the same molecular mass as the recombinant synexins expressed in Escherichia coli using cDNAs containing or lacking the cassette exon, respectively. The difference in size is consistent with the molecular masses predicted from the proteins encoded by the alternatively spliced synexin mRNAs. Another type of synexin mRNA contained a longer 3'-noncoding region generated by the selection of an alternate poly(A) signal. Northern analysis of human fibroblast RNA showed the presence of two bands (2.0- and 2.4-kilobase) when hybridized to a cDNA fragment of the coding region of synexin, but only the 2.4-kilobase band hybridized to a probe made from the longer 3' end.

Animals

Transplacental distribution of weak acids in mice: accumulation in compartments of high pH.

The transplacental distribution of three weak acids was measured in mice on gestational day 11. The ratio of drug concentration in maternal, embryonic, and extraembryonic compartments compared to maternal plasma followed a consistent hierarchy such that amniotic fluid greater than or equal to exocoelomic fluid greater than embryo greater than embryo plasma greater than maternal skeletal muscle. This distribution pattern correlates well with pH in these compartments and suggests that pH gradients between compartments are an influential factor in determining weak acid drug disposition during pregnancy.

Acids

Genomic organization and chromosomal localization of the human nucleolin gene.

Nucleolin, a eukaryotic nucleolar phosphoprotein, is involved in the synthesis and maturation of ribosomes. To characterize the genomic organization and regulatory sequences of this gene, two overlapping lambda clones containing the human nucleolin gene plus flanking regions were isolated from a genomic library using human nucleolin cDNA. Southern blots of genomic DNA from human, several mammals, chicken, and yeast revealed that the nucleolin gene is well conserved across these species. The gene consists of 14 exons with 13 intervening sequences and spans approximately 11 kilobases of DNA. Analysis of the splice junctions indicated that the amino-terminal domain and the four RNA binding domains plus the nuclear localization signal are split into adjacent exons. Sequences from the 5'-flanking and the first intron contain a high content of GC residues which is consistent with nucleolin being a "housekeeping" gene. Promoter elements include an atypical TATA box (GTTA), one CCAAT box much further from the initiation site, three reverse compliments of CCAAT (ATTGG), and two pyrimidine-rich nucleotide stretches. In addition, this region and the first intron contain numerous potential Sp1, GCF, CRE-fos, GCN, AP-1, AP-2, UCE, and sequences similar to the glucocorticoid receptor binding site. The transcription start site was determined by primer extension and S1 nuclease mapping of RNA from human liver. One Kpn and three Alu repeats were found within two of the middle introns. The 3'-untranslated portion of the gene contains five homology blocks in a 100-base pair region that are highly conserved among human, mouse, and hamster genomes. Finally, we have determined that the human nucleolin gene is located on chromosome 2q12-qter and is present at one copy per haploid genome. A restriction fragment length polymorphism with EcoRI has been detected in the gene.

Amino Acid Sequence

Circadian periodicity in microfilarial counts.

We propose a second-order harmonic model to describe circadian periodicity in the 24-h cycle of microfilarial counts. Rhesus monkeys (Macaca mulatta) were infected with Brugia malayi. Definite evidence of circadian periodicity was found in the microfilarial counts. The estimates of the parameters, periodicity indices and peak hours were determined.

Analysis of Variance

Cloning and sequencing of the human nucleolin cDNA.

A cDNA containing the entire coding region for human nucleolin has been isolated from a lambda gt10 human retinal library using a bovine cDNA probe. The cDNA hybridized to a transcript of 3000 bases from fast-dividing cells, as well as terminally differentiated tissues of several species. Translation of the nucleotide sequence revealed a long open reading frame which predicts a 707 amino acid protein with several distinct domains. These include repeating elements, four conserved RNA-binding regions, a glycine-rich carboxy-terminal domain and sites for phosphorylation, glycosylation and dibasic cleavage. Human and bovine nucleolin exhibited more additions and/or substitutions of aspartate, glutamate and serine residues in the chromatin-binding domains by comparison with the hamster and mouse nucleolins. These differences may be related to species-specific functions in transcription.

Adrenal Medulla

Calcium channel activity of purified human synexin and structure of the human synexin gene.

Synexin is a calcium-dependent membrane binding protein that not only fuses membranes but also acts as a voltage-dependent calcium channel. We have isolated and sequenced a set of overlapping cDNA clones for human synexin. The derived amino acid sequence of synexin reveals strong homology in the C-terminal domain with a previously identified class of calcium-dependent membrane binding proteins. These include endonexin II, lipocortin I, calpactin I heavy chain (p36), protein II, and calelectrin 67K. The Mr 51,000 synexin molecule can be divided into a unique, highly hydrophobic N-terminal domain of 167 amino acids and a conserved C-terminal region of 299 amino acids. The latter domain is composed of alternating hydrophobic and hydrophilic segments. Analysis of the entire structure reveals possible insights into such diverse properties as voltage-sensitive calcium channel activity, ion selectivity, affinity for phospholipids, and membrane fusion.

Amino Acid Sequence

Studies on in vitro survival of Isoparorchis hypselobagri.

Adult parasites, Isoparorchis hypselobagri were maintained in vitro in four basic salt solutions, viz. Stephenson saline, AB saline, Tyrode's solution and Locke's solution. Survival was prolonged by adding sugars to Tyrode's solution. Of all the sugars, glucose was found most effective and parasites survived for 39 days in solution containing glucose, 6 days in galactose and sucrose, 10 days in fructose and lactose and 12 days in maltose. The parasites absorb carbohydrates through the cuticle. Ligated parasites survived for a longer period than unligated parasites. The pH 9 was considered as an optimum for maximum survival of the parasites Isoparorchis hypselobagri for 39 days.

Absorption

Protein metabolism in Isoparorchis hypselobagri.

Isoparorchis hypselobagri was maintained in a non-nutrient medium containing streptomycin and penicillin to prevent bacterial growth for a period of 12 hours. The parasites were kept under aerobic conditions at a suitable temperature of 30 degrees C. The protein content in Isoparorchis hypselobagri ranged from 56.25% to 67.5% with an average of 63.5%+/-4.6 of dry seight of the tissue. The nitrogen of Isoparorchis hypselobagri excreted as ammonia, after 12 hours of starvation was 3.04% of the total body nitrogen and as uric acid 1.6% of total body nitrogen.

Aerobiosis

Amino acid composition in Isoparorchis hypselobagri.

The free amino acids of Isoparorchis hypselobagri have been determined by the chromatographic method (two-dimensional ascending). These are: leucine, valine, proline, alanine, glycine, glutamic acid and aspartic acid.

Amino Acids

Egg-shell formation in Isoparorchis hypselobagri.

Egg-shell formation in Isoparorchis hypselobagri is descirbed. The egg-shell in Isoparorchis hypselobagri is colorless and transparent as in case of Diplodiscus mehrai (Madhavi, 1968). Eleven histochemical tests were used to elucidate the process and it was observed that the basic mechanism of egg-shell formation followed the quinone tanned system as all the three precursors i.e. proteins, phenols and phenolase were present in vitelline globules.

Animals