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Biomedical subjects

A Das

Publications and source records attributed to A Das.

At least 19 recordsLinked to original sources
PubMed2001

Solution structure of a guanine-N7-linked complex of the mitomycin C metabolite 2,7-diaminomitosene and DNA. Basis of sequence selectivity.

2,7-Diaminomitosene (2,7-DAM), the major metabolite of the antitumor antibiotic mitomycin C, forms DNA adducts in tumor cells. 2,7-DAM was reacted with the deoxyoligonucleotide d(GTGGTATACCAC) under reductive alkylation conditions. The resulting DNA adduct was characterized as d(G-T-G-[M]G-T-A-T-A-C-C-A-C) (5), where [M]G stands for a covalently modified guanine, linked at its N7-position to C10 of the mitosene. The adducted oligonucleotide complements with itself, retaining 2-fold symmetry in the 2:1 drug-duplex complex, and provides well-resolved NMR spectra, amenable for structure determination. Adduction at the N7-position of G4 ([M]G, 4) is characterized by a downfield shift of the G4(H8) proton and separate resonances for G4(NH(2)) protons. We assigned the exchangeable and nonexchangeable proton resonances of the mitosene and the deoxyoligonucleotide in adduct duplex 5 and identified intermolecular proton-proton NOEs necessary for structural characterization. Molecular dynamics computations guided by 126 intramolecular and 48 intermolecular distance restraints were performed to define the solution structure of the 2,7-DAM-DNA complex 5. A total of 12 structures were computed which exhibited pairwise rmsd values in the 0.54-1.42 A range. The 2,7-DAM molecule is anchored in the major groove of DNA by its C10 covalently linked to G4(N7) and is oriented 3' to the adducted guanine. The presence of 2,7-DAM in the major groove does not alter the overall B-DNA helical structure. Alignment in the major groove is a novel feature of the complexation of 2,7-DAM with DNA; other known major groove alkylators such as aflatoxin, possessing aromatic structural elements, form intercalated complexes. Thermal stability properties of the 2,7-DAM-DNA complex 5 were characteristic of nonintercalating guanine-N7 alkylating agents. Marked sequence selectivity of the alkylation by 2,7-DAM was observed, using a series of oligonucleotides incorporating variations of the 5'-TGGN sequence as substrates. The selectivity correlated with the sequence specificity of the negative molecular electrostatic potential of the major groove, suggesting that the alkylation selectivity of 2,7-DAM is determined by sequence-specific variation of the reactivity of the DNA. The unusual, major groove-aligned structure of the adduct 5 may account for the low cytotoxicity of 2,7-DAM.

Alkylating Agents↗
PubMed2001

Model-independent source imaging using two-pion correlations in (2 to 8)a GeV Au+Au collisions.

We report a particle source imaging analysis based on two-pion correlations in high multiplicity Au+Au collisions at beam energies between 2A and 8A GeV. We apply the imaging technique introduced by Brown and Danielewicz, which allows a model-independent extraction of source functions with useful accuracy out to relative pion separations of about 20 fm. The extracted source functions have Gaussian shapes. Values of source functions at zero separation are almost constant across the energy range under study. Imaging results are found to be consistent with conventional source parameters obtained from a multidimensional Hanburg-Brown-Twiss analysis.

Journal Article↗
PubMed2001

Selective activation of mitomycin A by thiols to form DNA cross-links and monoadducts: biochemical basis for the modulation of mitomycin cytotoxicity by the quinone redox potential.

Mitomycin A (MA) but not mitomycin C (MC) cross-linked linearized (32)P-pBR322 DNA in the presence of dithiothreitol (DTT) or glutathione (GSH), as shown by a sensitive DNA cross-link assay. Incubation of calf-thymus DNA with MA and DTT or mercaptoethanol (MER) resulted in the formation of MA-DNA adducts, which were isolated from nuclease digests of the drug-DNA complexes by HPLC. The adducts were characterized by their UV absorption spectra, electrospray ionization mass spectrometry (ESIMS), and facile conversion from 7-methoxy- to 7-amino-substituted mitosene type adducts upon 10% NH(4)OH treatment, which were identical with known adducts of MC. Both DNA interstrand and intrastrand cross-link adducts, linking two deoxyguanosine residues at N(2), as well as several deoxyguanosine-N(2) monoadducts of MA, were identified. No DNA adducts were formed with MC under the same conditions. A specificity of DNA cross-link formation for the CpG sequence was observed using 12-mer synthetic oligodeoxyribonucleotides as substrates and as DNA sequence models, in analogy to the known CpG sequence specificity of MC-induced DNA cross-links. MA is known to be more cytotoxic by 2-3 orders of magnitude than MC, and this property correlates with redox potentials of MA (-0.19 V) and MA analogues that are higher than those of MC (-0.40 V) and its analogues. It is suggested that the biochemical basis for the higher cytotoxic potency of MA is MA's propensity to be reductively activated by cellular thiols while MC is resistant to thiol activation. This distinction is probably derived from the large difference between the quinone redox potentials of the two drugs.

Ammonium Hydroxide↗
PubMed2001

Differential role of cytosolic phospholipase A2 in the invasion of brain microvascular endothelial cells by Escherichia coli and Listeria monocytogenes.

Invasion of brain microvascular endothelial cells (BMECs) is a key step in the pathogenesis of meningitis due to Escherichia coli and Listeria monocytogenes. Although host cell actin cytoskeletal rearrangements are essential in BMEC invasion by E. coli K1 and L. monocytogenes, the underlying signaling mechanisms remain unclear. This study demonstrates that host cell cytosolic phospholipase A2 (cPLA2) contributes to E. coli K1 invasion of BMECs but not to L. monocytogenes invasion of BMECs. This difference was observed with 4-bromophenacyl bromide, a nonselective PLA2 inhibitor, and arachidonyl trifluoromethyl ketone, a selective cPLA2 inhibitor, and was confirmed with BMEC derived from cPLA2 knockout mice. Activation of cPLA2 leads to generation of intracellular arachidonic acid, which is metabolized via cyclooxygenase (COX) and lipo-oxygenase (LOX) pathways into eicosanoids. COX and LOX inhibitors also significantly inhibit E. coli K1 invasion of BMECs.

Alkaloids↗
PubMed2001

Zonal flow and zonal magnetic field generation by finite beta drift waves: a theory for low to high transitions in tokamaks.

The understanding of low to high (L-H) transition in tokamaks has been an important area of investigation for more than a decade. Recent 3D finite beta simulations of drift-resistive ballooning modes in a flux tube geometry by Rogers et al. [Phys. Rev. Lett. 81, 4396 (1998)] have provided a unique parametrization of the transition in a two-dimensional phase space. Comparison of the threshold curve in this phase space with data from ASDEX and C-MOD has shown very good agreement. In this Letter we provide a simple theory, based on the generation of zonal flow and zonal magnetic field in a finite-beta plasma, which explains this threshold curve for L-H transition in tokamaks.

Journal Article↗
PubMed2001

Model for interacting instabilities and texture dynamics of patterns.

A simple model to study interacting instabilities and textures of resulting patterns for thermal convection is presented. The model, consisting of a twelve-mode dynamical system derived for periodic square lattice, describes convective patterns in the form of stripes and patchwork quilt. The interaction between stationary zigzag stripes and standing patchwork quilt pattern leads to spatiotemporal patterns of twisted patchwork quilt. Textures of these patterns, which depend strongly on Prandtl number, are investigated numerically using the model. The model also shows an interesting possibility of a multicritical point, where stability boundaries of four different structures meet.

Journal Article↗
PubMed2001

Abrogation of tumor induced Ly49 expression on mouse spleen cells by mitomycin C.

Mouse spleen cells were activated with IL2 for 4 days in the presence or absence of paraformaldehyde fixed YAC (PFY) tumor cells (spleen cell: PFY ratio 100:1). Fresh spleen cells had poor expression of Ly49A but the expression was significantly upregulated by IL2 activation. Addition of PFYs resulted in a further boosting of Ly49A expression. Besides Ly49A, the Ly49C receptors were also upregulated by PFYs. Upregulated Ly49 expression was not restricted to NK cells (NK1.1 positive cells) but was also seen on T cells (TCRbeta positive cells). Time kinetics studies indicated that maximum upregulation of Ly49 in response to PFY cells occurred on day 3 and 4 and the expression declined thereafter. Ly49 expression in response to PFYs was completely blocked if spleen cells were pre-treated with Mitomycin C, an inhibitor of DNA synthesis. These results show that the addition of a small number of paraformaldehyde fixed YAC cells to spleen cell cultures undergoing IL2 activation, resulted in a significant upregulation of Ly49 receptors and this process was dependent upon cell proliferative activity.

Animals↗
PubMed2001

The Leptomonas seymouri spliced leader RNA promoter requires a novel transcription factor.

The spliced leader RNA gene promoter in Leptomonas seymouri requires three promoter elements for efficient and accurate transcription of the spliced leader RNA. The upstream most element appears to have a functional homolog in Leishmania species and in the African trypanosomes. The protein factor, promoter binding protein-1, interacts with the upstream element and appears to function as a basal transcription factor. Promoter binding protein-1 has three subunits; 36, 41 and 57 kDa. Using microsequencing techniques, we have obtained peptide sequence from each subunit. These data have enabled us to recently identify the Leptomonas gene that encodes the 41 kDa subunit. The 41 kDa subunit, comprised of 381 amino acids, is a founding member of a new class of transcription factors since extensive database searches revealed no homology to any known protein. This subunit, encoded by a single copy gene, has a potential nuclear localisation signal at amino acid positions 71-76. There are also multiple dileucine repeats with unknown function. Anti-41 kDa protein polyclonal antibodies are being employed to test the function of the 41 kDa subunit in PBP-1 activities.

Amino Acid Sequence↗
PubMed2001

Characterisation of the gene encoding type II DNA topoisomerase from Leishmania donovani: a key molecular target in antileishmanial therapy.

The gene encoding type II DNA topoisomerase from the kinetoplastid hemoflagellated protozoan parasite Leishmania donovani (LdTOP2) was isolated from a genomic DNA library of this parasite. DNA sequence analysis revealed an ORF of 3711 bp encoding a putative protein of 1236 amino acids with no introns. The deduced amino acid sequence of LdTOP2 showed strong homologies to TOP2 sequences from other kinetoplastids, namely Crithidia and Trypanosoma spp. with estimated identities of 86 and 68%, respectively. LdTOP2 shares a much lower identity of 32% with its human homologue. LdTOP2 is located as a single copy on a chromosome in the 0.7 Mb region in the L.donovani genome and is expressed as a 5 kb transcript. 5'-Mapping studies indicate that the LdTOP2 gene transcript is matured post-transcriptionally with the trans-splicing of the mini-exon occurring at -639 from the predicted initiation site. Antiserum raised in rabbit against glutathione S-transferase fusion protein containing the major catalytic portion of the recombinant L.donovani topoisomerase II protein could detect a band on western blots at approximately 132 kDa, the expected size of the entire protein. Use of the same antiserum for immunolocalisation analysis led to the identification of nuclear, as well as kinetoplast, antigens for L.donovani topoisomerase II. The in vitro biochemical properties of the full-length recombinant LdTOP2 when overexpressed in E.coli were similar to the Mg(II) and ATP-dependent activity found in cell extracts of L.donovani.

5' Untranslated Regions↗
PubMed2001

Directed flow of lambda hyperons in (2-6 )A GeV Au+Au collisions.

Directed flow measurements for Lambda hyperons are presented and compared to those for protons produced in the same Au+Au collisions (2A, 4A, and 6A GeV; b<5-6 fm). The measurements indicate that Lambda hyperons flow consistently in the same direction but with smaller magnitudes. A strong positive flow [for Lambdas] has been predicted in calculations which include the influence of the Lambda-nucleon potential. The experimental flow ratio Lambda/p is in qualitative agreement with expectations (approximately 2/3) from the quark counting rule at 2A GeV but is found to decrease with increasing beam energy.

Journal Article↗
PubMed2001

Profile of acetylcholinesterase in brain areas of male and female rats of adult and old age.

Inhibition of acetylcholinesterase (AChE)-metabolizing enzyme of acetylcholine, is presently the most important therapeutic target for development of cognitive enhancers. However, AChE activity in brain has not been properly evaluated on the basis of age and sex. In the present study, AChE activity was investigated in different brain areas in male and female Sprague-Dawley rats of adult (3 months) and old (18-22 months) age. AChE was assayed spectrophotometrically by modified Ellman's method. Specific activity (micromoles/min/mg of protein) of AChE was assayed in salt soluble (SS) and detergent soluble (DS) fractions of various brain areas, which consists of predominantly G1 and G4 molecular isoforms of AChE respectively. The old male rats showed a decrease (40-55%) in AChE activity in frontal cortex, striatum, hypothalamus and pons in DS fraction and there was no change in SS fraction in comparison to adult rats. In the old female rats the activity was decreased (25-40%) in frontal cortex, cerebral cortex, striatum, thalamus, cerebellum and medulla in DS fraction whereas in SS fraction the activity was decreased only in hypothalamus as compared to adult. On comparing with old male rats, old female rats showed increase in AChE activity in cerebral cortex, hippocampus and hypothalamus of DS fraction and decrease in hypothalamus of SS fraction. There was a significant increase in AChE activity in DS fraction of cerebral cortex, hippocampus, hypothalamus, thalamus and cerebellum in female as compared to male adult rats. However, no significant change in AChE activity was found in the SS fraction, except hypothalamus between these groups. Thus it appears that age alters AChE activity in different brain regions predominantly in DS fraction (G4 isoform) that may vary in male and female. These observations have significant relevance to age related cognitive deficits and its pharmacotherapy.

Acetylcholinesterase↗
PubMed2001

Fine-needle aspiration biopsy of metastatic soft-tissue sarcomas to lymph nodes.

This paper highlights the role of fine-needle aspiration cytology (FNAC) in 15 cases of metastatic soft-tissue sarcomas involving lymph nodes. Histopathology reports of the primary tumor were available in all cases. Histological diagnoses correlated well with the cytology reports. The most common type of sarcoma to involve the lymph node was embryonal rhabdomyosarcoma (6 cases), followed by synovial sarcoma (2 cases), leiomyosarcoma (2 cases), malignant fibrous histiocytoma (2 cases), fibrosarcoma (1 case), malignant peripheral nerve sheath tumor (1 case), and rhabdomyosarcoma (1 case). FNAC was thus helpful in the early diagnosis, proper staging, and management. Importantly, it obviated a lymph node biopsy in the majority of cases.

Adolescent↗
PubMed2001

Adhesion of the dissimilatory Fe(III)-reducing bacterium Shewanella alga BrY to crystalline Fe(III) oxides.

Shewanella alga BrY adhesion to hydrous ferric oxide, goethite, and hematite was examined. Adhesion to each oxide followed the Langmuir adsorption model. No correlation between adhesion and Fe(III) oxide surface area or crystallinity was observed. Zeta potential measurements suggested that electrostatic interactions do not influence S. alga BrY adhesion to these minerals. Cell adhesion does not appear to explain the recalcitrance of crystalline Fe(III) oxides to bacterial reduction.

Bacterial Adhesion↗
PubMed2001

Hybrid fuzzy logic committee neural networks for recognition of swallow acceleration signals.

Biological signals are complex and often require intelligent systems for recognition of characteristic signals. In order to improve the reliability of the recognition or automated diagnostic systems, hybrid fuzzy logic committee neural networks were developed and the system was used for recognition of swallow acceleration signals from artifacts. Two sets of fuzzy logic-committee networks (FCN) each consisting of seven member networks were developed, trained and evaluated. The FCN-I was used to recognize dysphagic swallow from artifacts, and the second committee FCN-II was used to recognize normal swallow from artifacts. Several networks were trained and the best seven were recruited into each committee. Acceleration signals from the throat were bandpass filtered, and several parameters were extracted and fed to the fuzzy logic block of either FCN-I or FCN-II. The fuzzified membership values were fed to the committee of neural networks which provided the signal classification. A majority opinion of the member networks was used to arrive at the final decision. Evaluation results revealed that FCN correctly identified 16 out of 16 artifacts and 31 out of 33 dysphagic swallows. In two cases, the decision was ambiguous due to the lack of a majority opinion. FCN-II correctly identified 24 out of 24 normal swallows, and 28 out of 29 artifacts. In one case, the decision was ambiguous due to the lack of a majority opinion. The present hybrid intelligent system consisting of fuzzy logic and committee networks provides a reliable tool for recognition and classification of acceleration signals due to swallowing.

Acceleration↗
PubMed2001

Female sex pheromone of brinjal fruit and shoot borer, Leucinodes orbonalis blend optimization.

The brinjal fruit and shoot borer, Leucinodes orbonalis is the major pest of eggplant in South Asia. Analysis of female pheromone gland extracts prepared from insects of Indian and Taiwanese origin confirmed (E)-11-hexadecenyl acetate (E11-16:Ac) as the major pheromone component with 0.8 to 2.8% of the related (E)-11-hexadecen-1-ol (E11-16:OH), as previously reported from Sri Lanka. The average quantity of E11-16:Ac extracted per female was estimated to be 33 ng, with a range of 18.9 to 46.4 ng when collected 2 to 3 hr into the scotophase. In field trials conducted in India, blends containing between 1 and 10% E11-16:OH caught more male L. orbonalis than E11-16:Ac alone. At the 1,000 microg dose, on white rubber septa, addition of 1% E11-16:OH to E11-16:Ac was found to be more attractive to male L. orbonalis than either 0.1 or 10% E11-16:OH. Trap catch was found to be positively correlated with pheromone release rate, with the highest dose tested, 3,000 microg, on white rubber septa catching more male moths than lower doses. Field and wind tunnel release rate studies confirmed that E11-16:OH released from white rubber septa and polyethylene vials at approximately twice the rate of E11-16:Ac and that the release rate of both compounds was doubled in polyethylene vials compared to white rubber septa. This difference in release rate was reflected in field trials conducted in Bangladesh where polyethylene vial dispensers caught more male moths than either black or white rubber septa, each loaded with the same 100:1 blend of E11-16:Ac and E11-16:OH in a 3,000 microg loading.

Animals↗
PubMed2001

Prevalence and natural history of subclinical hepatic encephalopathy in cirrhosis.

BACKGROUND AND AIMS: The natural history of subclinical hepatic encephalopathy (SHE) is unknown. The present study was conducted to study the prevalence and the natural history of SHE in patients with cirrhosis of the liver. METHODS: One hundred and sixty-five patients with cirrhosis of the liver were studied. A total of nine psychometric tests (trail making and Wechsler adult intelligence scale-performance (WAIS-P) tests) were administered. Subclinical hepatic encephalopathy was present if two or more psychometric tests were abnormal. Seventy-two patients (SHE 40, without SHE 32) also underwent serial psychometric testing on follow-up visits at 6-8 week intervals. RESULTS: Subclinical hepatic encephalopathy was present in 103 (62.4%) patients. The number and figure connection, block design and picture completion tests were the most useful in the detection of SHE. Severity of SHE, as assessed by the number of abnormal tests, was greater in patients with more severe liver disease. During follow up, SHE tended to persist or worsen in patients with poorer liver function. Although other clinical complications were similar in different groups, overt hepatic encephalopathy developed more commonly in those patients who had SHE at entry compared to those who did not (22.6 vs 5.6%, P = 0.044). Among the patients with SHE, the development of overt hepatic encephalopathy was more common in patients with Child's score of > 6 than with Child's score of <or= 6 (40 vs 5%, P = 0.019). CONCLUSIONS: We conclude that SHE is common in cirrhosis. The natural history of SHE is worse in patients with advanced cirrhosis and SHE probably predisposes the cirrhotic patient to overt hepatic encephalopathy.

Adult↗