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N K Verma

Publications and source records attributed to N K Verma.

26 records · Page 2Linked to original sources

Immunogenicity of the Shigella flexneri serotype Y (SFL 124) vaccine strain expressing cloned glucosyl transferase gene of converting bacteriophage SfX.

The glucosyl transferase gene (gtr) from bacteriophage phage X (SfX) caused partial conversion of serotype Y (group antigen 3, 4) to X (group antigen 7, 8) when introduced into a candidate vaccine strain of Shigella flexneri serotype Y (SFL124). The gtr gene caused conversion of O-antigens but did not eliminate the adsorption of the corresponding phage SfX. The hybrid strain expressing both group antigens 7, 8 and 3, 4 showed 75% protection when immunized guinea pigs were challenged with a wild-type S. flexneri serotype X strain. No protection was observed against serotype Y challenge, although group antigen 3, 4 was detected in the LPS of the hybrid strain. This suggests the importance of O-antigen immunity in the host defense against shigellosis.

Animals↗

Effects of intracervical injection of hyaluronidase in primigravidae during labour.

A study was conducted on 75 primigravidae in labour of which 50 were injected with intracervical injection of hyaluronidase and 25 were taken as control. Labour could be accelerated and shortened by an average of 1.95 hours (p < .01) after intracervical injection of hyaluronidase. Its effect was insignificant on the latent phase of labour while it significantly shortened the active phase of labour by 2.09 hours (p < .001). Its effect on maximum rate of cervical dilatation (0.85 cm/hour) was also found to be statistically significant ie, p < .001. Hyaluronidase had no effect on uterine contractions, duration of 2nd and 3rd stages of labour and on involution of the uterus. There was no incidence of cervical tear in any of the cases.

Cervix Uteri↗

Cloning and sequencing of the glucosyl transferase-encoding gene from converting bacteriophage X (SFX) of Shigella flexneri.

Shigella flexneri type Y strains (-;3,4) are converted to type X (-;7,8) by bacteriophage X (SFX) that causes glucosylation of the O-antigenic polysaccharide chain. The gene (gtr) encoding glucosyl transferase from bacteriophage X has been cloned and sequenced. The protein encoded by gtr consists of 416 amino acids with a M(r) of 47,369. The cloned gtr product was able to convert a S. flexneri strain type Y (SFL 124, a live attenuated candidate vaccine strain) to type X. The importance of the hybrid strain in vaccine development is discussed.

Amino Acid Sequence↗

Construction of aromatic dependent Shigella flexneri 2a live vaccine candidate strains: deletion mutations in the aroA and the aroD genes.

Live Shigella flexneri 2a vaccine candidate strains with deletion mutations in the aro genes were constructed. Tn10-generated auxotrophic mutations were transduced from Escherichia coli to S. flexneri 2a with bacteriophage P1CmCts. The tetracycline-sensitive derivatives of Tn10 mutants obtained were selected on Bochner's medium and checked by DNA-DNA hybridization using aroA and aroD gene specific probes. The vaccine candidate strains were tested to assess the efficacy of protection in guinea-pig conjunctival epithelia (Sereny test). The strains did not cause keratoconjunctivitis and exhibited significant protection in the challenge experiments. A candidate vaccine strain (delta aroD) showed 100% protection against 10(7) c.f.u. of wild type strain in the immunized guinea-pigs.

Animals↗

Molecular characterization of the O-acetyl transferase gene of converting bacteriophage SF6 that adds group antigen 6 to Shigella flexneri.

Bacteriophage SF6 antigenically converts Shigella flexneri serotype Y strains (-;3,4) to type 3b carrying group antigen 6,3,4 by means of an O-acetylation of the O-antigenic polysaccharide chain. The gene for O-acetyl transferase of bacteriophage SF6 has been cloned, identified and sequenced. The predicted O-acetyl transferase protein encoded by this gene was found to consist of 333 amino acids, (37,185 daltons) and to have some similarity with the galactose-1-phosphate uridylyltransferase protein of Escherichia coli. The gene has been shown to function in a live vaccine strain of S. flexneri Y type (delta aroD), making it a 3b type. The converted type 3b strain, SFL1104, was found to elicit significant protection against challenge by both wild-type serotypes 3b and Y in a guinea-pig keratoconjunctivitis model.

Acetylation↗

Relationships among the rfb regions of Salmonella serovars A, B, and D.

The O antigens of Salmonella serogroups A, B, and D differ structurally in their side chain sugar residues. The genes encoding O-antigen biosynthesis are clustered in the rfb operon. The gene rfbJ in strain LT2 (serovar typhimurium, group B) and the genes rfbS and rfbE in strain Ty2 (serovar typhi, group D) account for the known differences in the rfb gene clusters used for determination of group specificity. In this paper, we report the nucleotide sequence of 2.9 kb of DNA from the rfb gene cluster of strain Ty2 and the finding of two open reading frames which have limited similarity with the corresponding open reading frames of strain LT2. These two genes complete the sequence of the rfb region of group D strain Ty2 if we use strain LT2 sequence where restriction site data show it to be extremely similar to the strain Ty2 sequence. The restriction map of the rfb gene cluster in group A strain IMVS1316 (serovar paratyphi) is identical to that of the cluster in strain Ty2 except for a frameshift mutation in rfbE and a triplicated region. The rfb gene clusters of these three strains are compared, and the evolutionary origin of these genes is discussed.

Amino Acid Sequence↗

O-antigen variation in Salmonella spp.: rfb gene clusters of three strains.

The O antigens of Salmonella serogroups A, B, and D differ structurally in their side-chain sugar residue. These genes encoding O-antigen biosynthesis are clustered in the rfb operon. We report here the molecular cloning and analysis of the rfb operons of Salmonella paratyphi A (serogroup A) and S. typhi (serogroup D). The regions of DNA nonhomology between the rfb operons of these serogroup A, B, and D representatives are identified, and the evolutionary derivation of serogroup A from a serogroup D progenitor is discussed.

Antigenic Variation↗

Characterization of enterotoxin produced by four Yersinia enterocolitica strains of pig origin.

All four isolates of Yersinia enterocolitica and one isolate of Y. frederiksenii from pigs were found to be enterotoxigenic. Whole-cell preparations of Y. enterocolitica isolates did not induce any change in the rabbit ligated gut test after 6 and 18 h of inoculation, but Y. frederiksenii on the other hand showed a positive gut response at 18 h. Cell-free supernatant (CFS) of all five isolates induced dilatation in the rabbit gut up to 6 h, after which Y. enterocolitica became negative, while Y. frederiksenii continued to show a reaction up to 18 h. CFS of all five isolates were also found positive with the infant mouse test. Of the five isolates of Yersinia, three gave a positive reaction for the permeability factor on rabbit skin. Yersinia enterotoxin could be concentrated by methanol extraction. It was stable at 100 degrees C for 20 min and at 120 degrees C for 15 min. However, its activity was lost at low (2.0) and high pH (10.0). Enterotoxic preparations of Y. enterocolitica lost part of their enterotoxic activity upon dialysis.

Animals↗