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Biomedical subjects

S B Sharma

Publications and source records attributed to S B Sharma.

At least 55 records · Page 3Linked to original sources
PubMed1988

The effects of purified pertussis components and lipopolysaccharide on the results of the mouse weight gain test.

The effects of highly purified components of Bordetella pertussis, that is pertussis toxin (PT) and filamentous haemagglutinin (FHA), and of lipopolysaccharide (LPS) were studied in the active mouse weight gain test (MWGT). The PT when given alone or with other components in various combinations caused weight losses and deaths 2-3 days after inoculation but FHA was not toxic in the MWGT. When FHA was given with PT, the toxic effect of PT was reduced. The LPS caused weight losses at 24 h which decreased when LPS was given with PT. The toxic effects of PT as indicated by late deaths and late weight losses or failure to gain weight continued until 14 days after inoculation. The various components had similar effects on mouse weight gain in both LACA and NIH strains of mice. The doses of PT used in the MWGT caused marked leucocytosis but FHA and LPS did not. No agglutinins appeared in the sera of mice inoculated with various purified components. The components were thus pure and did not contain agglutinogens.

Agglutinins↗
PubMed1988

The reduction of animal usage by the application of indirect haemagglutination in the potency testing of diphtheria toxoid in combined vaccines.

Eight Adsorbed Diphtheria-Tetanus vaccines and 13 Diphtheria-Tetanus-Pertussis vaccines made by four different manufacturers were tested for the potency of the diphtheria components in guinea-pigs by the method of British Pharmacopoeia (1973). Two-hundred-and-ten guinea-pig sera consisting of ten sera related to each vaccine sample thus obtained were titrated for diphtheria antitoxin by indirect haemagglutination (IHA) and the conventional toxin neutralization (TN) tests. Statistical analysis of the results showed a good correlation between the titres obtained with the two tests. The potencies of the diphtheria components of various vaccines calculated from the antitoxin content of the respective guinea-pig sera titrated by the IHA test correlated significantly with the potencies obtained from the antitoxin content titrated by the routinely used TN test. The use of IHA in place of the TN test thus offers as an alternative that permits a reduction in animal usage.

Animal Testing Alternatives↗
PubMed1988

Studies on the optimal conditions for inactivation of Bordetella pertussis organisms with glutaraldehyde for preparation of a safe and potent pertussis vaccine.

The optimal conditions for inactivation of Bordetella pertussis organisms with glutaraldehyde for the production of a safe and potent whole cell pertussis vaccine were investigated. Two bacterial harvests from B. pertussis strain 10536 were treated with glutaraldehyde, each with 0.025, 0.05 and 0.1% concentrations of glutaraldehyde for 10, 60 and 120 min. The nine types of glutaraldehyde-inactivated pertussis vaccine (GIPV) and conventional heat-inactivated pertussis vaccine (HIPV) preparations made from two bacterial harvests were comparatively evaluated for the mouse weight gain test (MWGT), potency, and the histamine-sensitization (HS) and leucocytosis-promoting-factor (LPF) tests. The minimum period for killing the B. pertussis organisms with glutaraldehyde was greater than 10 min for 0.025%, 10 min for 0.05% and 5 min for 0.1% concentration. The average loss in opacity varied from 5 to 10% for GIPV preparations and was 14% for HIPV preparations. The GIPV preparations except those inactivated with 0.025% glutaraldehyde for 10 min (GIPV-A) were much less toxic than the HIPV preparations in the MWGT. The GIPV-A preparations did not pass the MWGT. The GIPV preparations were also much less toxic in HS and LPF tests than the HIPV preparation. The potency of GIPV preparations inactivated with 0.05% glutaraldehyde for 10 min (GIPV-D) was similar to that of HIPV preparations. The prolonged treatments with glutaraldehyde reduced the potency. The GIPV-D preparation with good potency and less toxicity was found to be inactivated with glutaraldehyde under optimal conditions. All the preparations were innocuous in the abnormal toxicity test.

Aldehydes↗
PubMed1987

The effects of different inactivating agents on the potency, toxicity and stability of pertussis vaccine.

The effect of heat (56 degrees C for 10 min), formaldehyde (0.1% at 37 degrees for 24h), glutaraldehyde (0.05% at room temperature for 10 min), thimerosal (0.02% at 37 degrees C for 24h), acetone-I (three treatments at room temperature) and acetone-II (three treatments at room temperature and fourth treatment at 37 degrees C), when used as inactivating agents in the preparation of pertussis suspension, was studied with regard to potency, toxicity and stability. Five batches each of Bordetella pertussis strains 134 and 509 were used for the study. The thimerosal inactivated pertussis (TIP) preparation was 1.5-2 times more potent than the heat inactivated pertussis (HIP) preparation. The potency values of the formaldehyde inactivated pertussis (TIP) and glutaraldehyde inactivated pertussis (GIP) preparations were similar to those of the HIP preparation, while the potencies of the acetone-I treated pertussis (A(I)TP) and acetone-II treated pertussis (A(II)TP) preparations were about half those of the HIP preparation. The FIP preparation was the least toxic showing maximum weight gain in the mouse weight-gain test (MGWT), while the TIP preparation did not pass the MWGT. The weight gains shown the GIP, A(I)TP and A(II)TP preparations were greater than those shown by the HIP preparation. The potency of pertussis component in the adsorbed diphtheria-pertussis-tetanus (DPT) vaccine was stable at 4-8 degrees C and 25 degrees C for three months for all types of pertussis vaccine. There was about 54-65% loss in the potency of the samples after three months at 35 degrees C. The inactivating agents used in the manufacture of pertussis preparations had no effect on the stability of the vaccine.

Acetone↗
PubMed1987

Glutaraldehyde inactivated pertussis vaccine: a less histamine sensitizing vaccine.

The effects of different inactivating agents on the biological activity of the histamine sensitization factor of Bordetella pertussis toxin were examined. The agents were used for inactivation in the preparation of whole cell pertussis suspension. The histamine sensitizing activity was reduced to 36.9-13.3% by treatment with glutaraldehyde, to about 50% by treatment with formaldehyde and by the acetone-II treatment, relative to the reduction by heat treatment. Treatment with thimerosal and the acetone-I treatment did not reduce the histamine sensitizing activity as the 50% histamine sensitizing doses of the heat inactivated pertussis preparation, the thimerosal inactivated pertussis preparation and the acetone-I treated pertussis preparation were very similar. Glutaraldehyde has thus been found to be a better inactivating agent for the preparation of a safe pertussis suspension as it considerably reduced the histamine sensitizing activity of pertussis toxin.

Acetone↗
PubMed1987

Glutaraldehyde inactivated pertussis vaccine: a safe vaccine in the innocuity test.

The adsorbed diphtheria-pertussis-tetanus (DPT) vaccine containing glutaraldehyde inactivated pertussis vaccine passed the innocuity test only after one month's storage of the preparation at 4-8 degrees C when the adsorbed DPT vaccine containing pertussis vaccine inactivated by heat, formaldehyde, thimerosal or acetone did not pass the test. The adsorbed DPT vaccine became innocuous when it was stored at 4-8 degrees C. The innocuity test was passed more easily in guinea pigs than in mice.

Animals↗
PubMed1987

Further observations on the stability of cholera vaccine at different temperatures.

The stability of cholera vaccine samples held at 4-8 and 20-25 degrees C for 2 1/2 years was studied with regard to the number of organisms, total nitrogen content and relative antigenicity. After this length of time, the percentage loss in the number of organisms averaged 10.7 at 4-8 degrees C and 22.7 at 20-25 degrees C. The maximum loss in number of organisms took place during the first six months and in the next two years there was very little loss in the number of organisms. The total nitrogen content after 2 1/2 years remained almost unaltered at these temperatures. All the ten samples had acceptable relative antigenicity for both the Inaba and Ogawa serotypes after exposure at 4-8 degrees C for 2 1/2 years, while at 20-25 degrees C only two samples for the Inaba serotype and one sample for the Ogawa serotype retained antigenicity within acceptable limits after the same time. The loss of antigenicity after 2 1/2 years at 20-25 degrees C in comparison with antigenicity at 4-8 degrees C ranged from 41.5 to 83.1% (mean 61.9%) for the Inaba serotype and from 47.3 to 80.3% (mean 65.1%) for the Ogawa serotype. From this study on the cholera vaccine manufactured at the Central Research Institute, Kasauli, it is concluded that the shelf life of cholera vaccine may be increased from 18 months to at least 24 months, when the vaccine is stored under refrigeration.

Animals↗
PubMed1987

A comparison of the indirect haemagglutination test with the toxin neutralization test for the estimation of diphtheria antitoxin in mouse sera.

Serum samples from 42 groups of mice immunized for different immunization periods with various doses of Adsorbed Diphtheria-Tetanus Vaccine, Adsorbed Diphtheria-Tetanus and Pertussis Vaccine and a standard diphtheria toxoid were assayed for their diphtheria antitoxin content by indirect haemagglutination (IHA) and by toxin neutralization (TN) tests. A very good correlation of 0.91 was obtained between the results of the two methods. There was no statistically significant difference between the IHA and the TN titres obtained. Adsorption with sheep red cells and treatment of the sera with 2-mercaptoethanol had no effect on the IHA titres. The minimum level of antitoxin detectable by the IHA test was 0.00039 IU ml-1. IHA proved to be a sensitive, specific and reproducible method which can be used reliably for the assay of diphtheria antitoxin in mouse sera.

Animals↗
PubMed1987

Evaluation of repeated immunoassays (mouse intracerebral potency tests) of the second International Standard of Pertussis Vaccine.

During one year 27 potency assays of second International Standard of Pertussis Vaccine were performed, when the Indian Standard of pertussis vaccine was being standardized. In all the 27 assays, the 50% immunizing dose (ImD50) was calculated both by Wilson-Worcester method and by probit analysis. The ImD50 of the International Standard varied from 0.022 I.U. to 0.076 I.U. (Mean 0.041 I.U.) when it was calculated by Wilson-Worcester method and from 0.0226 I.U. to 0.0704 I.U. (Mean 0.0402 I.U.) when it was calculated by probit analysis. There were no statistically significant differences between ImD50 values calculated by Wilson-Worcester method and by probit analysis. The challenge dose of Bordetella pertussis in terms of LD50 units had no effect on the ImD50 values. The average slope (b) of the assays was 1.92 when calculated by probit analysis and the average homogeneity factor (alpha) in these assays calculated by Wilson-Worcester method was 1.73.

Animals↗
PubMed1986

Nucleotide sequence of Rhizobium meliloti RCR2011 genes involved in host specificity of nodulation.

A 6 kb DNA segment of the R. meliloti 2011 pSym megaplasmid, which contains genes controlling host specificity of root hair infection and of nodulation, was cloned and sequenced. The DNA sequence analysis, in conjunction with previous genetic data, allowed identification of four nod genes designated as E, F, G and H. nodH is divergently transcribed with respect to nodFE and nodG. A conserved nucleotide sequence was found around 200 bp upstream of the translation start of nodF, nodH and nodA. This sequence is also present upstream of common nodA and species specific nodF genes of other Rhizobium species. The predicted protein products of nodF and nodG show homology with acyl carrier protein and ribitol dehydrogenase, respectively. The nodH product contains a rare sequence of four contiguous proline residues. Comparison with the nod gene products of R. leguminosarum shows that species specific nodFE products are as well conserved as those of common nodABC and nodD genes.

Amino Acid Sequence↗
PubMed1986

Effects of elevated temperatures on the opacity and toxicity of pertussis vaccines manufactured with different inactivating agents.

Plain pertussis vaccine manufactured with different inactivating agents was evaluated for its stability at 4-8, 25 and 35 degrees C with regard to the opacity and the toxicity [mouse weight gain test (MWGT) and histamine sensitizing (HS) activity]. Two pools each of heat inactivated pertussis vaccine (HIPV), formaldehyde inactivated pertussis vaccine (FIPV), glutaraldehyde inactivated pertussis vaccine (GIPV), thimerosal inactivated pertussis vaccine (TIPV) and acetone (I) treated pertussis vaccine [A(I)TPV] were taken for the study. The test for determination of opacity and MWGT were performed at monthly intervals for three months and the test for HS activity was performed after three months exposure of the samples at various temperatures. There was almost no loss in the opacity of GIPV and TIPV at all the selected storage temperatures and of HIPV and FIPV at 4-8 degrees C, while A(I)TPV lost about 16% opacity in three months at 4-8 degrees C. At 25 degrees C after three months, the percentage loss in opacity of HIPV averaged about 12, of FIPV 17 and of A(I)TPV 33. At 35 degrees C after three months, the average percentage loss in opacity of HIPV was 16, of FIPV was 25 and of A(I)TPV was 33. Regarding the effect of elevated temperatures on the toxicity of the vaccine, it was observed that the samples held at 35 degrees C for three months were the least toxic followed by those held at 25 and 4-8 degrees C as detected by MWGT and the test for HS activity.

Acetone↗
PubMed1984

Effect of storage temperatures on opacity and total nitrogen content of cholera vaccine.

Fluid cholera vaccine was examined for its stability at 4-8 degrees C, 20-25 degrees C and 37 degrees C with regard to the number of organisms present and the total nitrogen content. Ten batches of fluid cholera vaccine manufactured at the Central Research Institute, Kasauli, India were taken for the study. The number of organisms was determined at weekly intervals for the first month and at monthly intervals for the next 11 months. The percentage loss in the number of organisms after one year averaged 6.5 at 4-8 degrees C, 20.6 at 20-25 degrees C and 21.5 at 37 degrees C. The maximum loss in number of organisms took place during the first six months at these temperatures, in the following six months the number of organisms remained almost constant even at 37 degrees C. The total nitrogen content of the vaccine remained virtually unaltered during the same period. From this study it was concluded that the number of organisms and the total nitrogen content does not reflect the antigenicity nor potency of the vaccine.

Cholera Vaccines↗