Sabita Majumdar

Use of RNA arbitrarily primed-PCR fingerprinting to identify Vibrio cholerae genes differentially expressed in the host following infection.

ABSTRACT Evidence suggests that a repertoire of Vibrio cholerae genes are differentially expressed in vivo, and regulation of virulence factors in vivo may follow a different pathway. Our work was aimed at characterization of in vivo-grown bacteria and identification of genes that are differentially expressed following infection by RNA arbitrarily primed (RAP)-PCR fingerprinting. The ligated rabbit ileal loop model was used. The motility of in vivo-grown bacteria increased by 350% over that of in vitro-grown bacteria. Also, the in vivo-grown cells were more resistant to killing by human serum. By using the RAP-PCR strategy, five differentially expressed transcripts were identified. Two in vitro-induced transcripts encoded polypeptides for the leucine tRNA synthatase and the 50S ribosomal protein, and the three in vivo-induced transcripts encoded the SucA and MurE proteins and a polypeptide of unknown function. MurE is a protein involved in the peptidoglycan biosynthetic pathway. The lytic profiles of in vivo- and in vitro-grown cells suspended in distilled water were compared; the former was found to be slightly less sensitive to lysis. Ultrathin sections of both cells observed under the transmission electron microscope revealed that in contrast to the usual wavy discontinuous membrane structure of the in vitro-grown cells, in vivo-grown cells had a more rigid, clearly visible double-layered structure. The V. cholerae murE gene was cloned and sequenced. The sequence contained an open reading frame of 1,488 nucleotides with its own ribosome-binding site. A plasmid containing the murE gene of V. cholerae was transformed into V. cholerae 569B, and a transformed strain, 569BME, containing the plasmid was obtained. Ultrathin sections of 569BME viewed under a transmission electron microscope revealed a slightly more rigid cell wall than that of wild-type 569B. When V. cholerae 569B and 569BME cells were injected separately into ligated rabbit ileal loops, the transformed cells had a preference for growth in the ileal loops versus laboratory conditions.

Intracellular Development of Choleraphage Φ149 under Permissive and Nonpermissive Conditions: An Electron Microscopic Study

Intracellular development of choleraphage phi 149 following infection of Vibrio cholerae and Vibrio eltor cells under different conditions was examined by thin-section electron microscopy. Degradation of the host DNA following infection and formation of mature phage particles inside the infected cells were demonstrated. The concatemeric DNA intermediate formed during intracellular replication of phi 149 DNA in permissive hosts was resolved. In confirmation of biochemical evidence, no concatemeric DNA intermediate was observed for infection in high-phosphate medium.

Ulltrastructural and trace metal studies on radiographers’ hair and nails

Scalp hair and fingernail samples of 42 medical radiographers and 42 nonradiographers (control) with matching age groups and food habits were collected for this study. Trace metal estimation by atomic absorption spectrometry (AAS) has indicated a significant increase (P < 0.001) in Zn, Cu, and Cd contents in the radiographers’ hair and nails. Scanning electron microscopy (SEM) reveled structural changes in the hair and nails of radiographers. Significant alterations in the Zn and Cd contents along with extensive structural damage in the hair and nails probably indicate that low-dose Χ-radiation imposes stress on these radiation workers.

Intracellular Replication of Choleraphage φ92

The broad host range choleraphage φ92 contains a linear double-stranded DNA molecule of 68 kilobase (kb) pairs with 3′ overhang cohesive ends. Following infection with phage φ92, the host macromolecular synthesis is shut off within the first 5 min, and synthesis of phage-specific DNA is detectable after 7 min of infection. At early times during infection, phage DNA is replicated in circular form and the synthesis is membrane associated. The closed circular form serves as the precursor for the synthesis of the mature phage DNA which is eventually packaged into the phage head. Pulse labeling of ultraviolet-irradiated infected cells at different times during infection has allowed identification of about 30 phage-specific proteins of which 6 are structural proteins. These proteins appear during the infection cycle in two distinct phases, early and late. Eighteen early and 12 late proteins have been identified.

A trace metal (zinc and iron) study on low dose x-radiation response in rat skin.

There is no reliable bio-dosimeter regarding low dose radiation effects in mammalian systems. In this study, chronic low dose (< 1 cGy) whole body x-irradiated rat skin have shown altered trace metal (zinc and iron) content which clearly indicated the redistribution of these metals in the integumentary system. The decreased zinc to iron ratios suggested enhanced oxidative stress of the tissue. Changes in trace metal content in irradiated rat skin, as a biological response to low dose radiation, were non-linear. Moreover, the lowered zinc content of E2, E3, E4 and E5 dose groups suggested a different steady state, compared to the control. The Zn: Fe ratio decreased with increasing radiation dose.

Phage φ92: A New Choleraphage Infecting Vibrio cholerae Cells Belonging to Serovars O1 and Non-O1

A new choleraphage, φ92, with an icosahedral head and no tail structure has been described. This phage has a wide host range. It infects and lyses strains belonging to different serovars and biotypes