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Biomedical subjects

S K Ghosh

Publications and source records attributed to S K Ghosh.

At least 127 records · Page 7Linked to original sources

A clonal CD4-positive T-cell line established from the blood of a patient with Sézary syndrome.

The reported inability to establish long-term T-cell lines from the blood of cutaneous T-cell lymphoma patients with circulating neoplastic T cells has hindered the development of an in vitro system to investigate Sézary syndrome. We have established a rapidly proliferating T-cell line from the peripheral blood of a patient with Sézary syndrome, which expresses a mature helper T-cell phenotype and contains cytogenetic abnormalities and T-cell receptor gene rearrangements identical to those in the patient's blood. The method of establishment and characteristics of this line are described.

Aged↗

Lack of dose-response effect of terfenadine on resting bronchomotor tone in patients with asthma.

Bronchodilatation data from three, four-period cross-over studies were combined to assess the effect of oral terfenadine 60, 120, 180 mg and placebo on the airways of 26 patients with atopic asthma. Meta-analysis of these data showed that mean changes in FEV1 from pre-dose to 4 hr were 11.4, 14.6 and 11.8% for the three doses of terfenadine, respectively, and -2.9% for placebo. There was a significant treatment effect (P = 0.0001) but no effect of dose. Terfenadine, a non-sedating histamine H1-receptor antagonist, caused bronchodilatation in a single dose. Whether this effect is sustained with long-term treatment requires further investigation.

Adolescent↗

Effect of cetirizine on exercise induced asthma.

The effect of oral and inhaled cetirizine, a potent and specific H1 receptor antagonist, was studied in patients with exercise induced asthma. Twelve patients (five male; mean age 35.2 years) were given oral placebo or cetirizine 10 mg twice daily for one week, double blind and in randomised order, and exercised on a treadmill for six to eight minutes at a submaximal work load two hours after the final dose. There was no significant change in baseline FEV1 after treatment and cetirizine failed to inhibit exercise induced bronchoconstriction (maximum falls in FEV1 28% and 27% of baseline). In a further eight patients (four male; mean age 40.8 years) the effect of 1 ml cetirizine (5 and 10 mg/ml) given through a Wright nebuliser was compared with that of placebo in a double blind trial. The fall in FEV1 after exercise was reduced after both concentrations of cetirizine by 15.2% of baseline after 5 mg/ml and by 10.2% after 10 mg/ml, compared with 23.7% after placebo. In two patients cetirizine had no effect. In a further study cetirizine (10 mg/ml) given by inhalation displaced the geometric mean PC20 histamine 13.1 fold to the right by comparison with placebo. The reason for the difference between the effects of oral and of inhaled cetirizine on exercise asthma is not clear but may be related to differences in local concentration in the airway.

Administration, Inhalation↗

Pregnancy-induced cytotoxicity of splenocytes against mammary tumor cells in rats.

Our earlier observations indicate the possibility of involvement of a 'host factor' in pregnancy-induced protection against mammary carcinogenesis. The present investigation is an attempt to determine if this "host factor" is of immunological nature. Rat mammary tumors induced by 7,12-dimethylbenz[a]anthracene were used as target, and splenocytes from parous rats of the same strain were used as effector cells. The parous rats were divided into two groups according to the time period after parturition. Group 1 contained rats which were 5-13 days after delivery, group 2 had rats 14 or more days after parturition. In vitro cytotoxicity was determined by incubating the tumor cells with spleen cells in the target:effector ratios of 1:3, 1:10 and 1:30. Both groups 1 and 2 showed significant lysis of mammary tumor cells. These results were confirmed by the trypan blue exclusion test. The results showed a significantly higher cytotoxic capability of the spleen cells from parous rats against mammary tumor cells as compared to spleen cells from age-matched nulliparous rats.

Animals↗

Melittin-induced conformational changes in human lens protein.

Circular dichroism and fluorescence measurements showed a reduced conformational order in proteins of a normal human lens when they were incubated in vitro with melittin, a bee venom peptide. Since melittin is also known to react with lipids to induce a breakdown of vesicular structure, the observed denaturation of water-soluble proteins of a human lens that developed a cataract due to multiple bee stings may be accounted for by the effects of melittin to some extent. The melittin-induced decrease of conformational order, as observed in our in-vitro studies could thus be of physiological significance.

Circular Dichroism↗

Prevalence of malaria and economic loss in two major iron ore mines in Sundargarh district, Orissa.

A cross-sectional study on malaria was undertaken in May 1989 in the settlements of Kalta and Barsuan iron ore mines situated in a hilly area of Sundargarh district. Fever surveys revealed slide positivity rates of 33.9% and 34.8% in Kalta and Barsuan, respectively. Malaria infection rates as recorded through mass blood surveys in the resident population were 23.5 and 13.8%, respectively. Children up to 9 years age suffered most from malaria and in the age group of 2-9 years 37.3%, children had enlarged spleens with an average enlarged spleen (AES) index of 1.7. P. falciparum was the most prevalent species followed by P. vivax and P. malariae. Malaria vectors viz., An. fluviatilis and An. culicifacies were present in high densities. The present paper also brings out the economic loss due to malaria to the mining industry.

Adolescent↗

Vaccination with membrane-associated idiotype provides greater and more prolonged protection of animals from tumor challenge than the soluble form of idiotype.

The purpose of this work was to compare the efficacy of immunizing mice with a soluble vs a cell-associated form of a tumor Ag. A murine B cell tumor (2C3), which displays an Id on its cell surface, grows progressively and gives rise to Id-negative tumor variants in nonimmunized animals. We previously reported that the tumor variants arise as a consequence of Id-specific T cell suppression of the Id+ tumor. The Id-specific effector T cells are CD4+, CD8-. Based upon these findings vaccination protocols have been designed and tested to determine whether the expansion of tumor-specific effector T cells would eliminate the Id+ tumors and prevent the subsequent generation of Id- tumor variants in vivo. MHC-restricted T cells typically recognize soluble Ag subsequent to modification by an APC, and APC may ultimately express a processed form of the Ag that is different from that expressed on the surface of the tumor cells. Based upon this assumption, the efficacy of immunizing mice with cell-associated 2C3 Id was compared to immunization with a soluble form of the same Id. Mice were immunized with either irradiated 2C3 cells or syngeneic spleen cells to which 2C3 protein was covalently linked. These immunization protocols provided a complete and lasting protection against a tumor challenge of up to 1 x 10(6) tumor cells. In contrast, most mice hyperimmunized with a soluble form of the Id did not survive this level of tumor challenge in spite of the production of significant levels of anti-Id antibodies. Mice immunized with the soluble form of the Id, which did survive, produced slowly progressing tumors expressing a 1000-fold less of the marker Id. These results illustrate the importance of understanding and properly exploiting a host's natural response to a tumor-specific Ag when designing effective immunization protocols for cancer therapy.

Animals↗

Substrate specificity for myelin basic protein-specific protein methylase I.

The substrate specificity of bovine brain myelin basic protein (MBP)-specific protein methylase I (S-adenosyl-L-methionine:protein-L-arginine N-methyltransferase, EC 2.1.1.23), which methylates arginine residues of protein, has been studied using various MBPs, several synthetic peptides and heterogeneous nuclear ribonucleoprotein complex protein (hnRNP). (1) Among MBPs from different species of brain, the carp MBP was found to be the best substrate for MBP-specific protein methylase I. This high degree of methyl acceptability is most likely due to the fact that carp MBP is not in vivo methylated at the arginine residue (Deibler, G.E. and Martenson, R.E. (1973) J. Biol. Chem. 248, 2387-2391) and that the methylatable amino acid sequence is present in this protein. (2) In order to study the minimum chain length of MBP polypeptide which functions as the methyl acceptor, several synthetic polypeptides whose sequences are identical to the region surrounding the residue 107 of bovine MBP (the in vivo methylation site) were synthesized. It was found that the hexapeptide, Gly-Lys-Gly-Arg-Gly-Leu (corresponding to residues 104-109 of bovine MBP), was the shortest methyl accepting peptide, while the tetrapeptide, Gly-Arg-Gly-Leu (corresponding to residues 106-109) was inactive as a substrate. (3) hnRNP protein is known to contain methylarginine at residue 193 (Williams, K.R., Stone, K.L., LoPresti, M.B., Merrill, B.M. and Plank, S.R. (1985) Proc. Natl. Acad. Sci. USA 82, 5666-5670) which is post-translationally modified. Thus, the RNP protein overproduced in Escherichia coli and therefore did not contain methylarginine was examined for its methyl acceptability. It was found that neither MBP-specific nor histone-specific protein methylase I could methylate this methylarginine-less RNP protein. This suggests a possible existence of a distinct protein methylase I specific for this nuclear protein.

Amino Acid Sequence↗

Goat testis calmodulin: purification and physicochemical characterization.

Calmodulin has been purified in large quantities from goat (Capra hiscus) testis. The procedure includes heat treatment, hydrophobic interaction chromatography, and gel filtration. Goat testis calmodulin closely resembles other mammalian testis calmodulin studied so far. The protein has an extinction coefficient value (E1%1cm) of 2.09 at 280 nm, a Stokes radius of 23.2 A at 0.15 M KCl, and a frictional ratio of 1.38. Ca2+, and Tb3+ binding studies demonstrate that the protein has four Ca2(+)-binding sites with a Kd of 52.5 microM. Goat testis calmodulin shows close similarity to other calmodulins in the amino acid composition and in demonstrating an altered migration on SDS/PAGE upon Ca2+ binding. The protein also exhibits anomalously high values for molecular weight and Stokes radius as determined from the analytical gel chromatography and a change in its elution volume with the change of salt concentration in the eluant. These results have been discussed in view of the recently available knowledge from the crystallographic studies of rat testis calmodulin.

Amino Acids↗

Quartan malaria--an investigation on the incidence of Plasmodium malariae in Bisra PHC, District Sundargarh, Orissa.

A longitudinal study on the incidence of P. malariae was taken up from September 1988 to December 1989 in Bisra block, District Sundargarh, Orissa covering 38,615 population, which is mainly tribal. The area is a known hard-core malarious region in the Garhjat hill range in eastern India. In this study, out of 22,217 blood smears examined through weekly active surveillance, 7362 (33.1%) were found malaria parasite positive. Out of the total positive cases, 82 (1.1%) were P. malariae. These occurred mostly (91.4%) in persons below 40 years of age and children below 9 years accounted for 36.6% of total quartan malaria cases. In this age group the disease was found to be associated with splenomegaly (average enlarged spleen 2.07; spleen rate 45.9%) and 9 out of 13 mixed infections of P. malariae with P. falciparum and/or P. vivax were detected from this age group. This is the first report of quartan malaria from this area.

Adolescent↗

Cloning and characterization of mutL and mutS genes of Vibrio cholerae: nucleotide sequence of the mutL gene.

The mutL and mutS genes of Vibrio cholerae have been identified using interspecific complementation of Escherichia coli mutL and mutS mutants with plasmids containing the gene bank of V. cholerae. The recombinant plasmid pJT470, containing a 4.7 kb fragment of V. cholerae DNA codes for a protein of molecular weight 92,000. The product of this gene reduces the spontaneous mutation frequency of the E. coli mutS mutant. The plasmid, designated pJT250, containing a 2.5 kb DNA fragment of V. cholerae and coding for a protein of molecular weight 62,000, complements the mutL gene function of E. coli mutL mutants. These gene products are involved in the repair of mismatches in DNA. The complete nucleotide sequence of mutL gene of V. cholerae has been determined.

Amino Acid Sequence↗

Generation of somatic variants of a B cell hybrid mediated by a non-cytolytic L3T4+ idiotype-specific T cell.

We previously reported that idiotype (Id)-loss, stable somatic variants of a B cell hybrid, 2C3E1, are generated both in vitro and in vivo, after interaction of the Id-positive tumor cells with autologous Id-specific effector T cells. The present investigation was undertaken to elucidate further the nature and functional characteristics of the effector T cells. We report here that the idiotype-specific cells mediating the generation of Id- tumor variants are Thy1+ L3T4+ Lyt-2- cells, which respond to specific idiotypic stimulation by secreting IL-2 in vitro. No IL-2 is secreted in response to unrelated Ig or an Id/Ig-2C3E1 tumor variant. Furthermore, the Id-specific T cells exert strong suppressive effects on the expression of 2C3E1 Ig and the effects can be reversed by blocking the L3T4+ T cells with monoclonal anti-L3T4 antibody in vitro during the initial 3 days of co-culture. After 4 days, the T cell-mediated suppression of the 2C3E1-Id is irreversible. In addition to the in vitro studies we have determined that the administration of anti-L3T4 mAb to mice just before priming with idiotype-bearing tumor cells also abrogates the suppressive effects of the idiotype primed spleen cells on Ig expression of 2C3E1. To study the Id-specific effector T cells in more detail we have generated functional Id-specific L3T4+ T cell lines. These T cell lines have been shown to recapitulate the generation of Id- tumor variants that we observed with Id-primed spleen cells. It is concluded that L3T4+, Id-specific Ts cells are responsible for the generation of somatic variants of the B cell hybrid 2C3E1 and that the induction or selection of these variants progresses from a reversible phase to an irreversible phase.

Animals↗

Immunospecific targeting of cytosine arabinonucleoside-containing liposomes to the idiotype on the surface of a murine B-cell tumor in vitro and in vivo.

A new tumor model is described that is suitable for the evaluation of antibody-directed drug-delivery protocols and a modification in the procedure for covalently coupling antibody to the surface of drug-containing liposomes is presented. These immunospecific liposomes containing cytosine arabinonucleoside (Ara-C) have been tested in vitro and in vivo for their ability to kill a B-cell tumor. The target of the immunospecific-Ara-C liposomes is the idiotype associated with an antigen-specific immunoglobulin receptor on the cell surface of a murine B-cell hybrid (2C3). Affinity-purified antibodies specific for the idiotype were covalently coupled to modified lipid on the surface of the large unilamelar liposomes containing drug. These liposomes were shown to kill idiotype-positive 2C3 cells in vitro, but not idiotype-negative variants of this same cell line. It was also established in vitro that the drug-containing liposomes were at least 40 times more efficient than free Ara-C in the killing of the tumor cells. The 2C3 tumor was also propagated in vivo following the i.p. administration of tumor cells. The tumor grew initially as multiple foci within the peritoneum and subsequently spread to the spleen. Tumor-bearing mice were treated either with free Ara-C or with immunospecific liposomes containing Ara-C. Tumor growth in the primary tumor nodules and in the spleen was monitored by the administration of bromodeoxyuridine to the tumor-bearing animals followed by the immunofluorescent staining of cells with a monoclonal anti-bromodeoxyuridine antibody to estimate the proportion of cells in S phase. Our data from five out of seven animal experiments shows that the immunospecific-Ara-C liposomes, but not free drug, reduced tumor growth in the spleen. However, neither the liposomes containing drug nor the free drug were able to alter the growth of the primary tumor nodules growing in the peritoneal cavity. These results suggest that immunospecific-Ara-C containing liposomes may be useful in conjunction with other cytoreductive protocols in controlling tumor growth or preventing the spread of the tumor to other sites, but that immunospecific-Ara-C containing liposomes by themselves are not likely to eliminate an established tumor in vivo. We also demonstrate here that the administration of immunospecific-Ara-C containing liposomes in an animal having high levels of circulating tumor-associated antigen (i.e., IgG containing the idiotype) represents a potential clinically relevant hazard which must be considered when designing antibody-directed drug-delivery protocols.

Animals↗

Detection of a large number of subgroup 1 human rotaviruses with a "long" RNA electropherotype.

The long or short electrophoretic migration patterns of group A human rotaviruses are linked to their subgroup antigenic specificities. Long pattern isolates usually belong to subgroup 2 (SG2) and short pattern to subgroup 1 (SG1). To date detection of only 4 isolates which do not follow this linkage, have been reported. In the present communication we report the detection of unusually large number (39 isolates) of long pattern human isolates with SG1 specificities.

Antibodies, Monoclonal↗

Three-dimensional reconstruction of human limbs from tomographic views.

The recently developed medical imaging technique of tomography can be used with amazing success to learn about the interior of a human body. Each tomograph, however, is limited to only one section of the specific human limb. A series of tomographs taken at irregular intervals and various angles would provide three-dimensional information of the interior of the limb. Currently available 3-D surface display algorithms have limitations, particularly when applied to clinically important image data requiring fast and flexible interactive analysis. In addition to the problem of computation time is the cost of specialized hardware. A new algorithm has been designed for use with three-dimensional medical images which attempts to overcome these limitations. A computer graphics system is described which reconstructs three-dimensional images from tomographic sectional data. This 3-D surface algorithm can be exploited in planning reconstructive bone surgery. An example which illustrates the versatility and speed of the new algorithm is presented.

Algorithms↗

Lack of umuDC gene functions in Vibrio cholerae cells.

Attempts to identify an umuDC analog, using interspecific complementation of Escherichia coli mutants with plasmids containing a gene bank of Vibrio cholerae, were not successful. The DNA from none of the vibrio species examined including marine vibrios hybridized to E. coli umuC and umuD gene sequences. These cells are not mutable by ultraviolet (UV) light and cannot Weigle-reactivate UV-irradiated choleraphages, suggesting that vibrios are deficient in the umuDC operon. This possibility is supported by the fact that when the plasmid pKM101 carrying the mucAB genes is introduced into V. cholerae cells, they acquire the UV-mutable phenotype and UV-irradiated choleraphages can be Weigle-reactivated.

Blotting, Southern↗

Fatty acids and cholesterol: effect on the interaction of theophylline with bovine serum albumin.

The binding of theophylline (Th, 11-840 microM) to bovine serum albumin (BSA, 10 microM) using microdialysis technique in the presence of fatty acids (2.5-50.0 microM) and cholesterol (20-500 nM) indicates that fatty acids and cholesterol inhibit the binding of Th to BSA. The maximum inhibition (90.5%) occurs in presence of acetic acid (AA) followed by lauric acid (LA, 83.3%), palmitic acid (PA, 72.2%), oleic acid (OA, 44.4%) and cholesterol (22.2%). Fatty acids and cholesterol also decrease the number of binding sites and the affinity for the binding of Th to BSA. Such a decrease is maximum in the presence of AA followed by LA, PA, OA and cholesterol. Complete abolition of the low affinity binding site in the presence of AA indicates that the low affinity binding is predominantly ionic in nature while the high affinity binding involves ionic and other type(s) of unidentified force(s). This makes high affinity binding stronger than low affinity binding.

Acetates↗