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Biomedical subjects

A Ghosh

Publications and source records attributed to A Ghosh.

At least 217 records · Page 12Linked to original sources

Subplate neurons and the patterning of thalamocortial connections.

The patterning of the cerebral cortex into functionally distinct domains relies on the formation of appropriate connections between the thalamus and the cortex during development. To identify the mechanisms that underlie cortical target selection by thalamic axons, we have examined the role of cellular interactions in the formation of connections between the lateral geniculate nucleus (LGN) and the visual cortex during development of the cat visual system. The morphology of LGN axons as they grow towards the visual cortex suggests that interactions within the subplate zone may be important in the development of geniculocortical connections. The requirement for subplate neurons in this process was examined by ablating subplate neurons underlying the visual cortex at various developmental stages. When subplate cells are deleted between E38 and E42, prior to target innervation by LGN axons, these axons fail to select the visual cortex as their correct target and instead grow past it, staying restricted to the white matter. Deletion of subplate cells at later stages, between P2 and P7, does not affect target selection, but instead it prevents the segregation of LGN axons into ocular dominance columns within layer IV of the cortex. The effects of subplate neuron ablation suggest that interactions between thalamic axons and subplate cells are of critical importance in the specification of thalamocortical connections during development.

Animals↗

Molecular analysis of rice ragged stunt oryzavirus segment 9 and sequence conservation among isolates from Thailand and India.

Nucleotide sequences of rice ragged stunt virus (RRSV) genome segment 9 (S9) from a Thai and an Indian isolate were determined. Both sequences are 1132 bp long, contain a single large open reading frame (ORF) spanning nucleotide residues 14 to 1027 and are capable of encoding a protein of 38.5K. The two isolates are 94.6% and 99.4% identical at the nucleotide and amino acid level, respectively. The authenticity of these coding sequences was confirmed by identifying a approximately 38K protein in the RRSV particle with an N-terminal amino acid sequence identical to that inferred from the S9 ORF. Furthermore, cDNA of S9 from each isolate incorporated into the bacterial expression vector pGEX3-X produced a fusion protein that reacted with antibodies raised against purified RRSV particles. Cleaving these fusion proteins with protease factor X liberated a approximately 38K polypeptide.

Amino Acid Sequence↗

Characterization of an in vitro transcription system from rinderpest virus.

An in vitro transcription system for rinderpest virus (RPV) is described. Ribonucleoprotein complexes isolated from RPV-infected Vero cells, human lung carcinoma cells, or detergent-disrupted purified virions synthesized authentic RPV mRNAs for the N, P, M. F and H genes as identified by dot blot hybridization analysis with individual cDNA clones. The relative abundance of the mRNAs synthesized in vitro decreased from the 3' end of the genome to the 5' end, very similar to that observed with measles virus transcription in vitro. The transcription by purified virions was stimulated three-fold by the addition of infected human lung carcinoma cell lysate, demonstrating the involvement of host factor(s) in mRNA synthesis.

Animals↗

Distinct roles for bFGF and NT-3 in the regulation of cortical neurogenesis.

To identify molecules that regulate the transition of dividing neuroblasts to terminally differentiated neurons in the CNS, conditions have been developed that allow the neuronal differentiation of cortical precursor cells to be examined in vitro. In these cultures, the proliferation of undifferentiated precursor cells is controlled by basic fibroblast growth factor (bFGF). The proliferative effects of bFGF do not preclude the action of signals that promote differentiation, since addition of neurotrophin-3 (NT-3) antagonizes the proliferative effects of bFGF and enhances neuronal differentiation. In addition, blocking NT-3 function with neutralizing antibodies leads to a marked decrease in the number of differentiated neurons, without affecting the proliferation of cortical precursors or the survival of postmitotic cortical neurons. These observations suggest that bFGF and NT-3, by their distinct effects on cell proliferation and differentiation, are key regulators of neurogenesis in the CNS.

Animals↗

Evidence for a weak adaptive response to alkylation damage in Vibrio cholerae.

Wild-type Vibrio cholerae cells, when adapted by a stepwise treatment with sub-lethal concentrations of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), acquired resistance to killing and mutagenesis by subsequent challenges with higher concentrations of MNNG. This was also seen in the rec isogenic strain indicating that the observed phenomenon was not due to the induction of SOS functions. Further, the adapted cells of both the wild-type and rec strains could reactivate lethally alkylated phages with equal efficiency. Increased resistance of adapted cells correlated with the induction of a 17-kDa DNA methyltransferase, capable of repairing O6-methylguanine lesions in DNA. This induced methyltransferase was found to be antigenically unrelated to the Escherichia coli methyltransferase (Ada protein) as determined by Western blotting with polyclonal antiserum raised against the E. coli protein. Even though no counterpart of the constitutively expressed methyltransferase (Ogt) of E. coli could be detected in V. cholerae, several lines of evidence pointed towards the presence of an E. coli alk A-like gene in the organism.

Adaptation, Physiological↗

Biotype traits and antibiotic susceptibility of Vibrio cholerae serogroup O1 before, during and after the emergence of the O139 serogroup.

Sixty-nine strains of Vibrio cholerae O1 isolated at different times were analysed to investigate if there were any differences among the O1 strains isolated before, during and after the advent of the O139 serogroup. Of the 69 O1 strains examined, 68 belonged to the Ogawa serotype while one belonged to the Inaba serotype. With the exception of one strain all other strains of V. cholerae O1 belonged to the eltor biotype. A single O1 strain isolated before the emergence of the O139 serogroup could not be classified as either eltor or classical biotype because it was resistant to both classical and eltor specific bacteriophages. Marked variations in the susceptibility to antibiotics of V. cholerae O1 isolated during the different periods were observed. In addition, strains of V. cholerae isolated after the epidemic of serogroup O139 in Calcutta showed an expanding R-type with resistance to a variety of drugs as compared to the O1 strains isolated before the advent of the O139 serogroup. From this study, it is clear that there is a substantial mobility in genetic elements of V. cholerae O1 which necessitates a continuous monitoring to keep abreast of the changing traits of the etiologic agent of cholera.

Agglutination Tests↗

Studies on the genesis of Vibrio cholerae O139: identification of probable progenitor strains.

Four lines of evidence suggest that the recent outbreak strains of Vibrio cholerae O139 could have emerged from serogroup O1 strains typified by isolates M01 and M0477 described in this paper, which are neither truly classical nor truly E1 Tor in their biotype attributes. Firstly, like all O139 isolates, these O1 strains, isolated in Madras during and before the O139 outbreak, were resistant not only to polymyxin B but also to all biotype-specific choleraphages, i.e. classical phage phi 149 and E1 Tor phages e4 and e5. Secondly, the restriction fragment pattern (RFP) polymorphism displayed by these strains for the cholera toxin (ctx) gene, were identical with those produced by O139 isolates but were different from those of O1 type strains, namely V. cholerae 569B (classical) and V. cholerae MAK757 (E1 Tor). Thirdly, all the O139 isolates and the two O1 isolates carried an identical large number of copies of cholera toxin gene in their chromosomes. Finally, the outer-membrane protein profiles of strains M01 and M0477 were identical to those of O139 isolates but were different from those displayed by strains 569B and MAK757.

Bacterial Outer Membrane Proteins↗

Presence of lysogenic phage in the outbreak strains of Vibrio cholerae O139.

Four outbreak strains of Vibrio cholerae O139 from endemic areas of India and Bangladesh were found to carry lysogenic phage(s). All of these phage(s) produced turbid plaques characteristic of lysogeny on V. cholerae MAK 757 (El Tor, Ogawa) cells as well as on their VcA-1 lysogens but were unable to infect V. cholerae 154 (classical) cells, the universal host for all classical phages. Colonies in the turbid plaques were O139 lysogens and these developed an auxotrophic requirement, mainly for purines suggesting the integration of the prophage into the host chromosome. The immunity profile of the O139 phage(s) was similar to that of phage alpha but differed in the sensitivity of the phage lysogen of V. cholerae MAK 757 to subsequent infection by phage beta.

Bacteriophages↗

Induction of heat shock response in Vibrio cholerae.

General properties of the heat shock response in Vibrio cholerae were examined. Enhanced or de novo synthesis of 24 proteins was observed upon heat shock from 30 degrees C to 42 degrees C in cells labelled with [35S]methionine. A similar response could also be induced by a rise in temperature from 30 degrees C to 37 degrees C. Of these heat shock proteins, two were determined to be homologues of GroEL and DnaK, based upon their immunological cross-reactivity with antibodies raised against the Escherichia coli proteins. Three proteins, of molecular sizes 38, 44 and 48 kDa, which were undetectable in the 30 degrees C grown culture, appeared de novo after the heat shock. As in other prokaryotic systems thermal induction of many of the proteins was transient, but both DnaK and GroEL remained induced for at least 28 min after heat shock. DNA hybridization studies revealed that genes analogous not only to dnaK and groEL but also to dnaJ of E. coli exist in V. cholerae. Heat shock induced thermotolerance in V. cholerae but made the cells more sensitive to UV radiation. Unlike in E. coli, however, heat shock had no effect on the progeny virus yield in V. cholerae.

Bacterial Proteins↗

Increased expression of LD1 genes transcribed by RNA polymerase I in Leishmania donovani as a result of duplication into the rRNA gene locus.

Eukaryotic protein-coding genes are generally transcribed by RNA polymerase II (Pol II), which has a lower transcription rate than that of Pol I. We report here the duplication of two LD1 genes into the rRNA locus and their resultant transcription by Pol I. The multigenic LD1 locus is present in a 2.2-Mb chromosome in all stocks of Leishmania spp. and is also present in multicopy 200- to 450-kb linear chromosomes or multicopy circular DNAs in over 15% of stocks examined. Genomic rearrangement in Leishmania donovani LSB-51.1 resulted in duplication of a 3.9-kb segment of LD1 containing two genes (orfF and orfG) and of a 1.3-kb segment from approximately 10 kb downstream into the rRNA gene repeat region of the 1.2-Mb chromosome. Short sequences (12 or 13 bp) common to the 2.2-Mb LD1 and 1.2-Mb rRNA loci suggest that this gene conversion occurred by homologous recombination. Transcription of the duplicated genes is alpha-amanitin resistant, indicating transcription by Pol I, in contrast to the alpha-amanitin-sensitive (Pol II) transcription of the genes in the 2.2-Mb LD1 locus. This results in higher transcript abundance than expected from the gene copy number in LSB-51.1 and in elevated expression of at least the orfF gene product.

Amanitins↗

Effect of cerebellar modulation on gastroduodenal PGE2 and 5-HT content of rat.

Nodular cerebellar lesion decreased PGE2 and 5-HT content of gastroduodenal tissue along with a decrease in enterochromaffin (EC) cell count. On the other hand, when nodular lesioned rats were subjected to vestibular stimulation, both PGE2 and 5-HT content of gastroduodenal tissue and EC cell count increased, suggesting nodular cerebellar influence on PGE2 and 5-HT content of gastroduodenal tissue.

Animals↗

In vitro antifungal susceptibility of Candida.

To develop a simple, standardised procedure for antifungal susceptibility of yeasts, disc diffusion test was evaluated in comparison to agar dilution and the macrobroth dilution procedure of the National Committee for Clinical Laboratory Standards USA. One hundred Candida strains isolated from clinical material were used against four antifungal agents (amphotericin B, 5-flucytosine, ketoconazole and fluconazole). Inoculum size of 10(6) yeasts/ml just failed to produce confluent colonies at 37 degrees C after 48 h on yeast nitrogen-glucose-agar with or without asparagine medium. The zones of inhibition were correlated with both broth and agar dilution minimum inhibitory concentrations (MICs). The results were found to correlate well using amphotericin B, 5-flucytosine and fluconazole (with broth dilution only). But there was no significant correlation using ketoconazole. Therefore, disc diffusion susceptibility testing of Candida strains appears to be generally applicable except for azoles where broth dilution test would confirm the resistant isolates.

Antifungal Agents↗

Ultrasound measurement of placental thickness to detect pregnancies affected by homozygous alpha-thalassaemia-1.

Homozygous alpha-thalassaemia-1, a common cause of hydrops fetalis, is usually diagnosed invasively. We measured placental thickness by ultrasound at 10-21 weeks' gestation in 231 at-risk pregnancies. 60 were affected (by DNA analysis). The sensitivity and specificity in detecting affected pregnancies at cut-off of mean placental thickness plus 2 SD before 12 weeks' gestation were 0.72 (95% CI 0.52-0.93) and 0.97 (0.9-1.0). After 12 weeks sensitivity increased to 0.95 (0.89-1.0) and by 18 weeks reached 1.0 (0.60-0.99) without change in specificity. Selection of pregnancies at risk by measurement of placental thickness will reduce the number of invasive diagnostic procedures.

Female↗

Plasma and erythrocyte copper level as an indicator of disease activity during malignancy.

Copper and zinc concentrations in erythrocytes, plasma and whole blood were determined in mice bearing spontaneous-, transplanted- and benezo[a]pyrene-induced-tumour. The transplanted tumours studied were Sarcoma 180, Ehrlich carcinoma, and Dalton's lymphoma. Copper concentration in the malignant-tumour-bearing mice showed significant increases in erythrocytes and plasma, when compared with their normal controls. However, the zinc concentrations, although depressed in the different constituents of blood, were not significant enough to warrant any attention. Utilization of erythrocyte copper level as an additional marker of cancer activity is discussed.

Animals↗