PubMed Health⌕ Search

Biomedical subjects

A Basu

Publications and source records attributed to A Basu.

At least 127 records · Page 7Linked to original sources
PubMed1997

Incorporation of T cell counterparts in the fusion partners for generation of human monoclonal antibodies from Staphylococcus aureus stimulated B lymphocytes.

B cell growth and differentiation into immunoglobulin secreting cells is controlled by various cytokines and cell to cell contact with T cells. Fusion partner for human hybridoma therefore should accommodate all or some of these signaling systems to overcome the unique situation of MHC incompatibility, need for specific growth factors simultaneously taking into consideration the downstream processing of the product for the clinical use. We have thus directed our efforts towards the development of a fusion partner which would not need Epstein-Barr virus transformation of B cells prior to fusion. A nontransforming mitogen, formalinized Staphylococcus aureus (FSTA) was used for stimulating human B cells. Successful production of human IgM monoclonal antibody was achieved by incorporating Jurkat-4 cells in existing mouse human heterohybrid through fusion of these cells followed by fusion with human B cells. To accommodate chromosomes of both T and B cells after fusion, human myeloid precursor cells KG1a, and to incorporate T cell, HuT78 cells were fused. CD34+ and CD4+ hybrid of KG1a and HuT 78 cells-434 AM-when used as fusion partner could allow secretion of MAbs, however growth potential was low. SP2/0 cells were then incorporated in 434 AM cells to give myeloma environment to fused human B cells. Rabies virus neutralizing human IgG MAb secreting clone was generated by fusing FSTA stimulated human B cells with this fusion partner.

Animals↗
PubMed1997

Investigation of buffalopox outbreaks in Maharashtra State during 1992-1996.

During 1992-96, outbreaks of buffalopox zoonosis were reported from different villages in Jalgaon, Dhule and Beed districts of Maharashtra State. In humans, pox lesions were observed on the hands whereas in affected buffaloes and cows the lesions were noticed mainly on the teats and udder. Twenty two virus strains were isolated from the skin scabs collected from infected humans and milch animals. Neutralizing antibodies were detected not only in the sera of affected humans but also in their contacts. Detection of antibodies in young individuals from endemic area, who were neither vaccinated for smallpox nor had any contact with buffaloes or history of any poxvirus disease, is suggestive of occurrence of subclinical infection. A few children who had no contact with infected animals also showed clinical manifestations with disseminated lesions on the face, arm and buttocks, and thus suspected to have acquired infection through their infected parents or other family members indicating a possible man to man transmission. Therefore, in the light of discontinuation of smallpox vaccination, buffalopox outbreaks need to be monitored carefully as this may emerge as a serious zoonotic disease in India.

Animals↗
PubMed1997

Investigation of the outbreak of cholera in Alleppey & Palghat districts, south India.

In May 1996, a massive outbreak of cholera occurred in Alleppey district of Kerala which spread to Palghat district by July 1996. Of the 575 patients hospitalized at the Alleppey Medical College hospital between May 1 and August 2, 1996, 30 deaths occurred with a case fatality rate of 5.2 per cent while of the 638 diarrhoea patients admitted at Agali PHC of Attapadi area in Palghat district, 30 (4.7%) deaths were recorded. Clinically, the patients had profuse watery diarrhoea with vomiting. The epidemic of cholera in Alleppey and Palghat districts was caused by V. cholerae O1 of the EITor biotype, Ogawa serotype which possessed both the ctxA and tcpA genes when examined by multiplex PCR. Gross contamination of water sources was incriminated as the cause of the epidemic.

Adolescent↗
PubMed1997

Comparison between the multiplex PCR, sensitivity to biotype specific phages & polymyxin B for biotyping of Vibrio cholerae O1.

A total of 196 Vibrio cholerae O1 strains isolated between 1970 and 1996 were biotyped by multiplex PCR, susceptibility to polymyxin B and sensitivity to biotype specific phages. We modified the multiplex PCR by increasing the primer concentration of tcpA to improve the results. Comparison of the results of modified multiplex PCR and sensitivity to biotype specific phages and to polymyxin B showed that multiplex PCR was as efficient as phage typing for biotyping of V. cholerae O1. All the strains of V. cholerae O1 could be accurately distinguished based on polymyxin B sensitivity. Thus our results show that susceptibility of strains of V. cholerae O1 to polymyxin B is the easiest method to biotype V. cholerae O1 and is feasible in most laboratories when compared with multiplex PCR and sensitivity to biotype specific phages.

Anti-Bacterial Agents↗
PubMed1997

A comparative study of silver binding nucleolar organiser regions (AgNORs) of breast lesions in histological sections and fine needle aspiration smears.

The study presents a comparative profile of AgNOR dot counting in different types of breast lesions in histopathological (HP) sections and fine needle aspiration cytology (FNAC) smears. The breast lesions chosen were non-neoplastic lesion like fibroadenosis, benign neoplastic lesion like fibroadenoma and malignant neoplastic lesion like infiltrating duct carcinoma-grade 2. The AgNOR counts of non-neoplastic lesion were significantly less in number than the neoplastic lesions--both benign and malignant, in both the HP section and FNAC smear. But the counts did not show significant difference in the two neoplastic lesions eg, fibroadenoma and infiltrating duct carcinoma-grade 2, in both the HP section and FNAC smear. The appearance of the dots, as felt by the observers, were more discriminating between the three lesions, eg, uniform small compact centrally placed in fibroadenosis; mostly uniform small compact but occasional large irregular in fibroadenoma and large irregular marginally located in infiltrating duct carcinoma. Counting was easier and the appearance of the dots more easily discernible in FNAC smear than the HP section as the smear was monolayer and the malignant cells were easily detected from macrophages and stromal cells. But the tissue fluid or secretions or blood when present in the smear gave the smear a dirty background which was disturbing to the observers. Thus this AgNOR technique, when applied in HP section or FNAC smear, appears cost ineffective, lengthy and tedious procedure; did not offer absolute histochemical discriminant for malignancy from benignancy. But the shape and size distribution and appearance of the dots showing much variability in FNAC smear than the HP section, might be of some help in the diagnosis of malignancy and discriminating from benignancy.

Biopsy, Needle↗
PubMed1996

The role of an E box binding basic helix loop helix protein in the cardiac muscle-specific expression of the rat cytochrome oxidase subunit VIII gene.

We have characterized the rat gene for muscle-specific cytochrome oxidase VIII (COX VIII(H)) and mapped the distal promoter region responsible for transcription activation in C2C12 skeletal myocytes and H9C2 cardiomyocytes. In both cell types, the promoter elements responding to the induced differentiation of myocytes map to two E boxes, designated as E1 and E2 boxes with a core sequence of CAGCTG. Gel mobility shift analysis showed that both E1 and E2 box motifs form complexes with nuclear extracts from H9C2 cardiomyocytes that were supershifted with monoclonal antibody to E2A but not with antibody to myo-D. Extracts from induced and uninduced H9C2 cardiomyocytes yielded different gel mobility patterns and also different E2A antibody supershifts suggesting a difference in the DNA-bound protein complexes cross-reacting with the E2A antibody. Transcriptional activity of the promoter construct containing intact E boxes was inhibited by coexpression with Id in differentiated H9C2 cardiomyocytes. Our results show the involvement of an E box binding basic helix loop helix protein in the cardiac muscle-specific regulation of the COX VIII(H) promoter.

Animals↗
PubMed1996

Effects of a change in the pattern of insulin delivery on carbohydrate tolerance in diabetic and nondiabetic humans in the presence of differing degrees of insulin resistance.

While it is well established that people with non-insulin dependent diabetes mellitus have defects in both insulin secretion and action, the relative contribution of each to glucose intolerance is not known. Therefore, nondiabetic (lean and obese) and non-insulin dependent diabetes mellitus subjects were studied on two occasions. On each occasion, insulin secretion was inhibited with somatostatin and glucose was infused in a pattern and amount that mimicked the systemic delivery rate normally observed after ingestion of 50 g of glucose. Insulin also was infused so as to mimic postprandial insulin profiles observed in separate groups of diabetic and nondiabetic subjects after food ingestion. Glucose turnover was measured using the isotope dilution method. A delayed pattern of insulin delivery (i.e., a "diabetic" insulin profile) led to higher (P < 0.05) glucose concentrations in all groups; however, the effects were transient, resulting in only a modest increase in the integrated glycemic responses. An isolated defect in insulin action had little effect on peak glucose concentration; however, it prolonged the duration of hyperglycemia, leading to a 2.5-4.2-fold increase (P < 0.05) in the integrated glycemic response. A combined defect in the pattern of insulin secretion and action was additive rather than synergistic. Both defects caused hyperglycemia by altering suppression of endogenous glucose release and stimulation of glucose disposal. Whereas obese diabetic and nondiabetic subjects had comparable defects in glucose clearance, non-insulin dependent diabetes mellitus subjects also had defects in hepatic insulin action. Thus, abnormalities in the pattern of insulin secretion and action alone or in combination impair glucose tolerance. An isolated defect in insulin action has a more pronounced and prolonged effect than does an isolated change in the pattern of insulin secretion. Hepatic and extrahepatic insulin resistance results in marked and sustained hyperglycemia.

Blood Glucose↗
PubMed1996

Temporal shifts in traits of Vibrio cholerae strains isolated from hospitalized patients in Calcutta: a 3-year (1993 to 1995) analysis.

This study presents results of a surveillance on cholera conducted with hospitalized patients admitted to the Infectious Diseases Hospital, Calcutta, India, from January 1993 to December 1995. The O139 serogroup of Vibrio cholerae dominated in 1993 but was replaced by O1 as the dominant serogroup in 1994 and 1995. The isolation rate of V. cholerae non-O1 non-O139 did not exceed 4.9% throughout the study period, while the isolation rate of the O139 serogroup in 1994 and 1995 was below 9%. No temporal clustering of any non-O1 non-O139 serogroup was observed. With the exception of 1 strain, none of the 64 strains belonging to the non-O1 non-O139 serogroup hybridized with ctx, zot, and ace gene probes, while 97.3 and 97.7% of the O139 and O1 strains, respectively, hybridized with all the three probes. Multiplex PCR studies revealed that all the O1 strains belonged to the EIT or biotype. There was a progressive increase in the cytotoxic response on CHO and HeLa cells evoked by culture supernatants of strains of V. cholerae non-O1 non-O139 isolated during 1994 and 1995 compared with the response evoked by those isolated in 1993. Dramatic shifts in patterns of resistance to antibiotics between strains of V. cholerae belonging to different serogroups and within strains of a serogroup isolated during different time periods were observed. There was a discernible increase in the incidence of multidrug-resistant strains of V. cholerae O1 isolated in 1994 and 1995 compared with that in 1993. On the basis of the results of this study, we predict the possibility of newer variants of V. cholerae emerging in the future.

Drug Resistance, Multiple↗
PubMed1996

Fine needle aspiration cytology of ophthalmic lesions.

Forty cases of ophthalmic lesions were subjected to fine needle aspiration cytology for pre-operative diagnosis which included retinoblastomas (5 cases), inflammatory and neoplastic lesions of the eyelid and lacrimal glands (29 cases) and extra-ocular orbital lesions (6 cases) inclusive of 3 cases of rhabdomyosarcomas. The lesions were distributed in both the eyes almost in equal proportions. Histological confirmation was obtained in 85% of cases; false positive in only one case with malignant neoplasm of the eyelid and false negative also in only one case with inflammatory pseudotumour which were, histologically, proved to be adenoma of tarsal gland and lymphocytic lymphoma respectively.

Biopsy, Needle↗
PubMed1996

Characterization of the protein kinase C signal transduction pathway in cisplatin-sensitive and -resistant human small cell lung carcinoma cells.

Protein kinase C (PKC) influences cellular sensitivity to cis-diamminedichloroplatinum(II) (cDDP). We have investigated whether the PKC signal transduction pathway is affected during the development of cellular resistance to cDDP. Activators of PKC, such as phorbol 12,13-dibutyrate (PDBu), enhanced the sensitivity of human small cell lung cancer H69 cells to cDDP by 2-fold but had no effect on the sensitivity of cDDP-resistant H69 cells (H69/CP) to cDDP. The maximum sensitization was achieved with 10 nM PDBu and blocked by down-regulation of PKC with higher concentrations of PDBu (1 microM) or bryostatin 1 (0.1 microM). PKC activity was decreased significantly in H69/CP cells compared to the drug-sensitive variant. A similar reduction in PKC activity was noted in ovarian carcinoma 2008 cells that were resistant to cDDP. A modest decrease in PKC activity was also observed in etoposide-resistant H69 (H69/VP-16) cells but not in Taxol-resistant H69 cells or bleomycin-resistant human head and neck carcinoma A-253 cells. H69 cells expressed conventional PKC alpha and-beta, novel PKC delta, atypical PKC zeta and-iota, and novel/atypical PKC mu. A decrease in cPKC alpha and-beta and an increase in nPKC delta were associated with the cDDP-resistant phenotype. The abundance of aPKC zeta or-iota was unaffected. H69/ VP-16 cells also displayed a reduction in cPKC beta and an increase in nPKC delta. Taxol-resistant H69 cells had no alteration in the expression of any of the PKC isozymes. Thus, a reduction in cPKCs and an increase in nPKC may be associated with cDDP resistance.

Antineoplastic Agents↗
PubMed1995

Oncogenic transformation alters cisplatin-induced apoptosis in rat embryo fibroblasts.

We have investigated whether cellular transformation by different oncogenes alters their responsiveness to the chemotherapeutic agent cis-diamminedichloroplatinum(II) (cDDP). Rat-6 (R6) embryo fibroblasts transformed by v-myc, rc-mos (rearranged mouse c-mos), v-src and activated c-H-ras oncogenes displayed differential sensitivities to cDDP, and the IC50 values of cDDP decreased by approximately 3.0-, 1.2-, 2.0- and 1.5-fold, respectively. Over-expression of myc, but not ras, src or mos, induced apoptosis in R6 cells. Nevertheless, all oncogenes influenced cellular susceptibility to apoptosis triggered by cDDP as judged by the appearance of condensed chromatin, formation of apoptotic bodies and internucleosomal cleavage of genomic DNA to 180 bp multimers, yielding a "laddered" electrophoretic profile on agarose gel. There was a correlation between the time- and concentration-dependent enhancement of cDDP-induced DNA fragmentation and potentiation of cDDP sensitivity by various oncogenes. Oncogenic transformation led to a decrease in expression of the PKC epsilon isoform, and stable transfection of PKC epsilon in R6 cells prevented cDDP-induced apoptosis and protected cells against cDDP cytotoxicity. Our results indicate that the differential sensitivity of oncogene-transformed cells to cDDP may be associated with the ability of various oncogenes to influence cDDP-induced apoptosis.

Animals↗
PubMed1995

Comparison of protein kinase C activity and isoform expression in cisplatin-sensitive and -resistant ovarian carcinoma cells.

Cellular sensitivity to cis-diamminedichloroplatinum(II) (cDDP) can be regulated by protein kinase C (PKC) signal transduction pathway. Activators of PKC were shown to enhance the sensitivity of human ovarian carcinoma 2008 cells to cDDP. We have examined whether or not the PKC signal transduction pathway is affected during development of resistance by tumor cells to cDDP. A 2-fold decrease in PKC activity was observed in cDDP-resistant ovarian carcinoma 2008/C13*5.25 cells compared with the drug-sensitive 2008 cells. Subcellular distribution studies revealed a reduction in both cytosolic and particulate PKC activities in 2008/C13*5.25 cells. The pattern of PKC isoform expression was compared in cDDP-sensitive and -resistant cell lines by Western blot analysis with isoform-specific antibodies to PKC. The parental cells expressed PKC alpha, -epsilon, and -zeta isoforms. The abundance of PKC alpha decreased significantly in 2008/C13*5.25 cells, whereas the amount of PKC epsilon increased moderately in the resistant variant, with no alteration in PKC zeta content. Therefore, a reduction in PKC alpha and/or an increase in PKC epsilon expression may be associated with the drug-resistant phenotype.

Blotting, Western↗
PubMed1995

The epithelial/carcinoma antigen EGP-1, recognized by monoclonal antibody RS7-3G11, is phosphorylated on serine 303.

RS7-3G11 is a murine monoclonal antibody (MAb) raised against human non-small-cell lung carcinoma, and is under clinical evaluation. The epithelial/carcinoma antigen EGP-1, defined by RS7-3G11, was isolated and purified to homogeneity from a cervical carcinoma cell line, ME180. EGP-1 is a glycoprotein with an average molecular mass of 47.8 kDa. Metabolic labeling of the antigen with 32P-orthophosphate and subsequent immunoprecipitation with RS7-3G11 showed that it is a phosphoprotein. Phosphoamino acid analysis of the in vivo phosphorylated EGP-1 revealed that the phosphorylation is on serine. In vitro analysis with purified antigen demonstrated that protein kinase C, and not protein kinase A, is involved in phosphorylating the antigen in vitro. In vitro analysis indicated a stoichiometry of phosphorylation of 0.54 mole of phosphate per mole of EGP-1. Phosphoamino acid analysis and phosphopeptide mapping of the antigen phosphorylated in vitro by protein kinase C showed that phosphorylation occurred on a serine residue, specifically on serine 303, located in the cytoplasmic domain of EGP-1. Treatment of ME180 cells with phorbol ester increased the phosphorylation of EGP-1. The biological function of EGP-1 remains to be elucidated. In this report we elucidate an involvement of protein kinase C in phosphorylating EGP-1, which may signify a role for this antigen in signal transduction across the cell membrane.

Amino Acid Sequence↗
PubMed1995

Effects of radiolabeling monoclonal antibodies with a residualizing iodine radiolabel on the accretion of radioisotope in tumors.

The effect of using a "residualizing" iodine radiolabel, dilactitol-iodotyramine, for radioimmunolocalization of antibodies to tumors was investigated. This tracer is designed to be lysosomally trapped after catabolism of the labeled antibody. mAbs RS7 and RS11 were used for in vivo and in vitro studies on the uptake and retention of radioisotope into tumor cells. Both are murine IgG1 mAbs with pancarcinoma reactivity, which react with integral membrane glycoproteins. mAb RS7 has been shown to be relatively rapidly catabolized by the antigen-bearing cell line Calu-3, whereas RS11 is catabolized more slowly in the same cells. An 111In- or 88Y-p-isothiocyanatobenzyl-diethylenetriamine pentaacetic acid conjugate was also tested because these radiometals are known to be lysosomally trapped, and iodination via chloramine T was used to provide a baseline. In vitro, a substantial increase in retention of the label by cells was observed when the dilactitol-tyramine DLT- or 111In-labeled mAbs were used, and the improvement gained by the use of these residualizing labels was greater with the use of the rapidly catabolized mAb (RS7) than it was with the more slowly catabolized mAb (RS11). In biodistribution studies in nude mice bearing Calu-3 tumor xenografts, a dramatic improvement in the tumor accretion of the radiolabel was seen with the use of the 131I-labeled DLT- or 88Y-labeled mAbs. For example, at day 7 the percentage of injected dose/g in the tumor was 5.54 +/- 1.47% (SD), 38.06 +/- 8.04%, and 43.18 +/- 19.50% for the conventionally iodinated, DLT- and 88Y-labeled RS7, respectively. Dosimetry calculations performed on the biodistribution data predict increases of approximately 8- and 4-fold in the absorbed dose to tumor with the use of 131I-labeled DLT- and 90Y-labeled mAbs, respectively, compared to the conventional 131I. In contrast to in vitro findings, these results were similar for both RS7 and RS11, suggesting that the use of DLT may be advantageous for most of the mAbs binding to the cell surface, including antibodies that are catabolized relatively slowly. The advantage of 131I-labeled DLT over 90Y is due to the longer physical half-life of the 131I.

Adenocarcinoma↗