PubMed Health⌕ Search

Biomedical subjects

J B Doshi

Publications and source records attributed to J B Doshi.

5 recordsLinked to original sources
PubMed2003

Comparative quantitation for the protein content of diphtheria and tetanus toxoids by DC protein assay and Kjeldahl method.

DPT, a combination vaccine against diphtheria, tetanus and pertussis is available since many years and still continued in the national immunisation schedule of many countries. Although highly potent, reactions to DPT vaccine are well known, mainly attributed to the factors like Pertussis component, aluminum adjuvant and lower purity of tetanus and diphtheria toxoids. The latter most important aspect has become a matter of concern, specially for the preparation of next generation combination vaccines with more number of antigens in combination with DPT. Purity of toxoid is expressed as Lf (Limes flocculation) per mg of protein nitrogen. The Kjeldahl method (KM) of protein nitrogen estimation suggested by WHO and British Pharmacopoeia is time consuming and less specific. Need has been felt to explore an alternative method which is quick and more specific for toxoid protein determination. DC (detergent compatible) protein assay, an improved Lowry's method, has been found to be much more advantageous than Kjeldahl method.

Chemistry Techniques, Analytical↗
PubMed1994

Transcriptional regulation of CD6 expression on human T lymphocytes by phorbol ester.

Activation of protein kinase C (PKC) has emerged as a major common signal-transducing mechanism for T cell activation and regulation of expression of T cell surface glycoproteins. Surface expression of the CD6 Ag is known to increase with T cell activation and CD6 itself may be involved in regulation of T cell activation. Therefore, we performed experiments to determine whether activation of PKC by phorbol ester induced an increase in CD6 expression and to investigate the mechanisms of such an effect. CD6 surface expression was up-regulated substantially in response to PMA on both mature and immature T cells, but only negligibly on B cell lines. This increase was blocked by PKC inhibitors. The PMA-induced increase in CD6 surface expression was accompanied by an increase in total CD6 protein, as detected by using Western blot analysis of whole cell lysates. After PMA stimulation, Northern blot analysis showed that steady state levels of CD6 mRNA increased in response to PMA treatment. This increase was blocked by cycloheximide, demonstrating that it was dependent on new protein synthesis. Nuclear run-on analysis showed that the rate of CD6 mRNA transcription increased by approximately two to threefold after PMA stimulation of Jurkat cells. Experiments in which we used actinomycin D showed that PMA had no significant effect on the t1/2 of CD6 mRNA. The data suggest that the effect of PMA on CD6 expression is mediated primarily by an increase in CD6 mRNA transcription after PKC activation. mAbs were used to determine whether augmented CD6 expression could be induced by perturbation of specific T cell surface molecules. Up-regulation of CD6 expression occurred when thymocytes were cultured with anti-CD2 Abs, but not with Abs to other functional T cell surface structures, and not when mature T cells were cultured with the anti-CD2 mAbs. Up-regulation of CD6 expression by activation of PKC, triggered in thymocytes by ligation of CD2, could allow CD6 to provide additional regulatory signals required for events in later stages of T cell activation and differentiation.

Antigens, CD↗
PubMed1993

Activation of human T cells through CD6: functional effects of a novel anti-CD6 monoclonal antibody and definition of four epitopes of the CD6 glycoprotein.

The CD6 glycoprotein is expressed primarily on lymphocytes and conveys co-activating signals to T cells, but its exact function and ligand(s) are unknown. A novel mAb, termed UMCD6, was demonstrated to recognize CD6 by immunoprecipitation, Western blotting, and reactivity with COS cells transfected with CD6 cDNA. UMCD6 was mitogenic for T cells and was strongly synergistic with phorbol ester in inducing T cell activation. UMCD6 enhanced the autologous mixed lymphocyte reaction as previously observed with another anti-CD6 mAb, anti-T12. The activating effects of UMCD6 were more striking than those of other anti-CD6 mAbs and encompassed all of the diverse stimulatory properties previously reported for other anti-CD6 reagents. However, neither UMCD6 nor other anti-CD6 antibodies alone or in combination with phorbol ester or IL-2 were able to induce thymocytes to proliferate. Stimulation by UMCD6 is dependent on accessory cell function in a manner not accounted for simply by antibody cross-linking. UMCD6 did not induce an increase in cytoplasmic free Ca2+, but the CD6 activation pathway appears to involve protein kinase C. UMCD6 and a panel of seven other anti-CD6 mAbs were used in a series of experiments to define four discrete epitopes of CD6 using the criteria of antibody cross-blocking, reactivity on reduced Western blots, and resistance to controlled V8 protease digestion. The functional mAbs UMCD6, 2H1, and anti-T12 each recognized a different epitope. Taken together, the results of these studies strongly reinforce the hypothesis that CD6 plays a significant and distinct role in T cell activation, and suggest that multiple regions of CD6 may be functionally active.

Antibodies, Monoclonal↗
PubMedDate not supplied

Schematic eye models for simulation of patient visual performance.

PURPOSE: To determine if model eyes can simulate the visual performance of normal human eyes under conditions of varying low myopic blur, pupil size, and contrast. METHODS: High and low contrast Bailey-Lovie logMAR visual acuity (VA) of three normal eyes of three subjects were measured for four artificial pupil sizes and ten levels of myopic defocus. Simulated visual acuities were then determined for three model eyes--the Indiana Eye with no spherical aberration, the Indiana Eye with average spherical aberration, and the Kooijman Eye--by generating optically aberrated VA charts for each testing condition using Visual Optics Lab software by Sarver and Associates, Inc, and having the subjects read high resolution printouts of these charts using a 3-mm pupil and optimal spectacle correction. The correlation between real VA and simulated VA was then plotted and a regression line calculated. RESULTS: Slopes for the Indiana Eye, Indiana Eye with spherical aberration, and Kooijman Eye were 0.98, 0.98, and 1.01 for high contrast, and 0.92, 0.67, and 0.75 for low contrast, respectively. The r2 values were 0.73, 0.74, and 0.77, for high contrast, and 0.69, 0.40, and 0.50 for low contrast, respectively. Under low contrast conditions the Indiana Eye VA was significantly closer to the real VA than that of the other two models (P<.0003). CONCLUSION: Visual performance can be simulated by eye models. The simple single surface Indiana Eye with no spherical aberration best modeled both high and low contrast visual acuity.

Adult↗