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Biomedical subjects

A Datta

Publications and source records attributed to A Datta.

At least 73 records · Page 4Linked to original sources
PubMed1997

X-ray reflectivity study of semiconductor interfaces.

The results of an X-ray reflectivity study of thick AlAs-AlGaAs and thin Ge-Si-Ge multilayers grown using metal-organic vapour-phase epitaxy and ion-beam sputtering deposition techniques, respectively, are presented. Asymmetry in interfaces is observed in both of these semiconductor multilayers. It is also observed that although the Si-on-Ge interface is sharp, an Si(0.4)Ge(0.6) alloy is formed at the Ge-on-Si interface. In the case of the III-V semiconductor, the AlAs-on-AlGaAs interface shows much greater roughness than that observed in the AlGaAs-on-AlAs interface. For thin multilayers it is demonstrated that the compositional profile as a function of depth can be obtained directly from the X-ray reflectivity data.

Journal Article↗
PubMed1997

A novel MAP-kinase kinase from Candida albicans.

A putative MAP-kinase kinase-encoding gene, CaSTE7, was isolated from Candida albicans by complementation of ste7 and ste11 mutants of the pheromone signal-transduction pathway of Saccharomyces cerevisiae. The nucleotide (nt) sequence revealed an ORF of 1767 nt encoding a putative protein of 589 amino acids (aa). CaSTE7 has a strong homology with MAP-kinase kinase STE7 of S. cerevisiae, the kinase domain having 45% homology with that of STE7. The deduced aa sequence contained all eleven consensus kinase subdomains found in MAP-kinase kinases. It can suppress the mating defect of ste5, ste11, ste7, and fus3 kss1 double mutants, but it cannot bypass the ste12 mutation. CaSTE7 behaves as a hyperactive allele of STE7, suppressing the mating defects of the pheromone signal-transduction pathway by constitutively stimulating STE12, and hence STE12-dependent processes.

Amino Acid Sequence↗
PubMed1997

Purification and characterisation of a hemolysin with phospholipase C activity from Vibrio cholerae O139.

A hemolysin was purified from a Vibrio cholerae O139 strain which moved as a single protein band of 67 kDa in SDS-PAGE. The hemolysin showed high level of phospholipase C activity. The purified phospholipase C-hemolysin demonstrated enterotoxic activity in rabbit ileal loop, suckling mice and enhanced permeability of rabbit skin. The pI of the purified hemolysin was 6.4. Erythrocytes from rabbit, chicken, guinea pig, sheep and horse were sensitive to the purified hemolysin in decreasing order of intensity. Erythrocytes from human and cow were unaffected by purified hemolysin.

Animals↗
PubMed1997

Influence of the dielectric property on microwave oven heating patterns: application to food materials.

Patterns of power absorption in a microwave oven for a range of dielectric properties of relevance to food processing were investigated. The governing Maxwell's equations with boundary conditions and a TE10 excitation were solved using a finite element method. Food properties were varied from values at their frozen state to values at high temperatures, as would be typical in a thawing process. For low-loss materials such as frozen foods, the high quality factor makes the heating significantly higher only when the size and shape of the load permit a dielectric cavity resonance in the load. Otherwise, the heating pattern will follow the modal electric field pattern of the oven. For moderate loss materials, the patterns will come from the modes of the dielectric cavity. The bandwidths of these modes are larger than the low-loss situation and their overlap results in a heating pattern that is somewhat more uniform. For high-loss materials, the concept of modes is no longer useful as the very large number of modes strongly overlap. The rapidly decaying field and power loss in the high-loss material can probably be characterized as an exponential decay.

Absorption↗
PubMed1997

The role of lysine-256 in the structure and function of sheep liver recombinant serine hydroxymethyltransferase.

The active site lysine residue, K256, involved in Schiff's base linkage with pyridoxal-5'-phosphate (PLP) in sheep liver recombinant serine hydroxymethyltransferase (rSHMT) was changed to glutamine or arginine by site-directed mutagenesis. The purified K256Q and K256R SHMTs had less than 0.1% of catalytic activity with serine and H4folate as substrates compared to rSHMT. The mutant enzymes also failed to exhibit the characteristic visible absorbance spectrum (lambda(max) 425 nm) and did not produce the quinonoid intermediate (lambda(max) 495 nm) upon the addition of glycine and H4folate. The mutant enzymes were unable to catalyze aldol cleavage of beta-phenylserine and transamination of D-alanine. These results suggested that the mutation of the lysine had resulted in the inability of the enzyme to bind to the cofactor. Therefore, the K256Q SHMT was isolated as a dimer and the K256R SHMT as a mixture of dimers and tetramers which were converted to dimers slowly. On the other hand, rSHMT was stable as a tetramer for several months, further confirming the role of PLP in maintenance of oligomeric structure. The mutant enzymes also failed to exhibit the increased thermal stability upon the addition of serine, normally observed with rSHMT. The enhanced thermal stability has been attributed to a change in conformation of the enzyme from open to closed form leading to reaction specificity. The mutant enzymes were unable to undergo this conformational change probably because of the absence of bound cofactor.

Animals↗
PubMed1996

Toward antibody-directed "abzyme" prodrug therapy, ADAPT: carbamate prodrug activation by a catalytic antibody and its in vitro application to human tumor cell killing.

Antibody-directed enzyme prodrug therapy, ADEPT, is a recent approach to targeted cancer chemotherapy intended to diminish the nonspecific toxicity associated with many commonly used chemotherapeutic agents. Most ADEPT systems incorporate a bacterial enzyme, and thus their potential is reduced because of the immunogenicity of that component of the conjugate. This limitation can be circumvented by the use of a catalytic antibody, which can be "humanized," in place of the bacterial enzyme catalyst. We have explored the scope of such antibody-directed "abzyme" prodrug therapy, ADAPT, to evaluate the potential for a repeatable targeted cancer chemotherapy. We report the production of a catalytic antibody that can hydrolyze the carbamate prodrug 4-[N,N-bis(2-chloroethyl)]aminophenyl-N-[(1S)-(1,3- dicarboxy)propyl]carbamate (1) to generate the corresponding cytotoxic nitrogen mustard (Km = 201 microM, kcat = 1.88 min-1). In vitro studies with this abzyme, EA11-D7, and prodrug 1 lead to a marked reduction in viability of cultured human colonic carcinoma (LoVo) cells relative to appropriate controls. In addition, we have found a good correlation between antibody catalysis as determined by this cytotoxicity assay in vitro and competitive binding studies of candidate abzymes to the truncated transition-state analogue ethyl 4-nitrophenylmethylphosphonate. This cell-kill assay heralds a general approach to direct and rapid screening of antibody libraries for catalysts.

Aniline Mustard↗
PubMed1996

Role of cell-associated N-acetyl-D-glucosamine specific haemagglutinin in the adhesion of Vibrio cholerae O1 to rabbit intestinal epithelial cells in vitro.

Previously a N-acetyl-D-glucosamine specific cell-associated haemagglutinin (HA) had been purified from a Vibrio cholerae O1 strain. This study documents the role of this purified HA as an adhesin of V. cholerae O1. A significant inhibition in the adhesion of V. cholerae O1 bacterial cells to isolated rabbit intestinal brush borders (RIBB) was observed when the latter were pretreated with purified HA in ELISA. Antibody raised against purified HA and Fab (IgG) fragment of this serum inhibited adhesion of the bacteria to isolated rabbit intestinal epithelial cells (RIEC). V. cholerae O1 (both Ogawa and Inaba serovars) showed less adherence to isolated RIEC of animals immunised with the purified HA. Patients convalescing from V. cholerae O1 infection showed high ELISA titres against the purified HA indicating that it is expressed in the host during the disease process.

Acetylglucosamine↗
PubMed1996

Destabilization of simple repetitive DNA sequences by transcription in yeast.

Simple repetitive DNA sequences in the eukaryotic genome frequently alter in length. In wild-type strains, we find that transcription through a repetitive poly GT tract destabilizes the tract four- to ninefold. In mismatch repair-deficient yeast strains, simple repeats are very unstable. High levels of transcription in such strains destabilize repetitive tracts an additional two- to threefold.

DNA Damage↗
PubMed1996

Electrophoretic mobility and immune response of outer membrane proteins of Vibrio cholerae O139.

The outer membrane (OM) protein components of a Vibrio cholerae O1 and four V. cholerae O139 strains, collected from cholera patients, were analysed by SDS-PAGE. A protein of 69 kDa molecular mass was observed only when the OMPs were prepared from strains grown in synthetic broth. As a result of passage in the rabbit ileal loop (RIL), virulence was enhanced, and a protein component around 18 kDa of the V. cholerae O139 OM became the major protein component. On immunoblot analysis with rabbit antiserum against V. cholerae O139 OM, it was shown that, apart from the major protein component of V. cholerae O1 OM of around 45 kDa and that of V. cholerae O139 OM of around 38 kDa, all other minor protein components were cross-reactive between the two serogroups. In immunoblot assays with convalescent sera obtained from V. cholerae O139-infected patients, it was observed that in addition to the lipopolysaccharide (LPS)-induced antibody, only the 38 kDa major protein component elicited considerable levels of antibody in the patient. Minor OM components of 18 kDa were detected in the immunoblot analysis by LPS-directed antibody, however, as the OM proteins are known to be associated with LPS.

Animals↗
PubMed1996

Purification of a cell-associated hemagglutinin from Shigella dysenteriae type 1.

A cell-associated hemagglutinin (HA) was isolated and purified from a clinical isolate of Shigella dysenteriae type 1 by affinity chromatography on a fetuin-agarose column. The purified hemagglutinin produced a single-stained protein band of around 66 kDa in sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). In an immunodiffusion test, HA-antisera produced a single precipitin band against the purified HA without exhibiting any reactivity towards lipopolysaccharide (LPS) of S. dysenteriae type 1 strain. Inhibition of the hemagglutination by the glycoproteins fetuin, asialofetuin and a sugar derivative N-acetyl-neuraminic acid but not by simple sugars, suggested the specific requirement of complex carbohydrate for binding. Electron micrographs of the purified HA revealed a morphology typical of globular protein.

Cells↗
PubMed1996

Mitotic crossovers between diverged sequences are regulated by mismatch repair proteins in Saccaromyces cerevisiae.

Mismatch repair systems correct replication- and recombination-associated mispaired bases and influence the stability of simple repeats. These systems thus serve multiple roles in maintaining genetic stability in eukaryotes, and human mismatch repair defects have been associated with hereditary predisposition to cancer. In prokaryotes, mismatch repair systems also have been shown to limit recombination between diverged (homologous) sequences. We have developed a unique intron-based assay system to examine the effects of yeast mismatch repair genes (PMS1, MSH2, and MSH3) on crossovers between homologous sequences. We find that the apparent antirecombination effects of mismatch repair proteins in mitosis are related to the degree of substrate divergence. Defects in mismatch repair can elevate homologous recombination between 91% homologous substrates as much as 100-fold while having only modest effects on recombination between 77% homologous substrates. These observations have implications for genome stability and general mechanisms of recombination in eukaryotes.

Base Sequence↗
PubMed1996

Antibody mediated inhibition of adhesion of Shigella dysenteriae 1 to guinea pig colonic epithelial cells in vitro.

Immunoglobulin G (IgG) fractions prepared from sera of Shigella dysenteriae 1 infected patients during the acute and convalescent phases of illness were found to be effective inhibitors of adhesion of S. dysenteriae cells to guinea pig colonic epithelial cells in vitro. The adhesion could also be inhibited by whole sera, their IgG fractions, and Fab fractions prepared from sera of rabbits immunized with whole bacteria, the outer membrane (OM) and lipopolysaccharide. The adhesive capability was best inhibited by the Fab fragment of antisera of rabbits immunized with whole bacteria.

Animals↗
PubMed1995

Association of increased spontaneous mutation rates with high levels of transcription in yeast.

Complex processes such as transcription, replication, repair, and recombination require changes in chromatin structure and the interactions of numerous trans-acting factors with DNA sequences, raising the possibility that these processes may be interrelated. Here the effect of transcription on the rate of spontaneous mutation in the yeast Saccharomyces cerevisiae was examined. With the use of a lys2 frameshift allele under the control of a highly inducible promoter, the rate of spontaneous reversion was shown to increase when the mutant gene was highly transcribed. Thus, transcriptionally active DNA and enhanced spontaneous mutation rates are associated in yeast.

Alleles↗
PubMed1995

cDNA cloning, overexpression in Escherichia coli, purification and characterization of sheep liver cytosolic serine hydroxymethyltransferase.

A sheep liver cDNA clone for the cytosolic serine hydroxymethyltransferase (SHMT) was isolated and its nucleotide sequence determined. The full-length cDNA of SHMT was placed under the control of T7 promoter in pET-3C plasmid and expressed in Escherichia coli. The overexpressed enzyme, present predominantly in the soluble fraction, was catalytically active. The recombinant SHMT was purified to homogeneity with a yield of 10 mg/l bacterial culture. The recombinant enzyme was capable of carrying out tetrahydrofolate-dependent and tetrahydrofolate-independent reactions as effectively as the native enzyme. The Km values for serine (1 mM) and tetrahydrofolate (0.82 mM) were similar to those of the native enzyme. The recombinant enzyme had a characteristic visible spectrum indicative of the presence of pyridoxal 5'-phosphate as an internal aldimine. The apoenzyme obtained upon removal of the cofactor was inactive and could be reconstituted by the addition of pyridoxal 5'-phosphate demonstrating that the recombinant SHMT was functionally very similar to the native SHMT. This overexpression of eukaryotic tetrameric SHMT in E. coli and the purification and characterization of the recombinant enzyme should thus allow studies on the role of specific amino acids and domains in the activity of the enzyme.

Amino Acid Sequence↗
PubMed1995

Composition-based prediction of dielectric properties of foods.

Prediction of accurate dielectric property data from fundamental principles for systems as complex as foods has not been possible. Simple prediction models based on easily measurable composition data can serve many useful purposes. Literature dielectric data on foods and their composition were statistically correlated. Dielectric data on salt solutions were measured to explain some of the results. When composition data were not available, standard handbook compositions were used. Inclusion of all types of foods (meats, fruits, and vegetables) inhibited any useful correlation with composition. Based on a smaller data set of meats, both dielectric constant and loss increased with water and salt content. Dielectric constant generally decreased with temperature whereas dielectric loss decreased with temperature at lower salt concentrations and increased with temperature at higher salt concentrations.

Electric Conductivity↗