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Biomedical subjects

G Das

Publications and source records attributed to G Das.

At least 37 records · Page 2Linked to original sources
PubMed2000

Immunization of cross-bred cattle against Hyalomma anatolicum anatolicum by purified antigens.

Extracts prepared from unfed larvae of Hyalomma anatolicum anatolicum were purified by immunoaffinity chromatography using anti-gut IgG as ligand. Affinity purified antigen (Aff-GHLAg) was used to immunize cross-bred (Bos taurus x B. indicus) calves of 6-7 months of age. Immunized calves rejected 70.6% larvae, 54.5% nymphs and 61.9% adults. No significant changes in the engorged weight of females was observed; however, significant decrease in the engorgement weight of larvae and nymphs was recorded. There was a significant decrease in the emerging nymphs (p < 0.05) and adults (p < 0.01) of the tick stages fed on immunized animals. SDS-PAGE analysis revealed three antigenic proteins of 100, 59.4 and 37 kDa responsible for induction of resistance in the host.

Animals↗
PubMed2000

Th1-specific bystander costimulation imparts resistance against Mycobacterium tuberculosis infection.

The protection against Mycobacterium tuberculosis infection is mediated by T helper type-1 (Th1) cells. Infection of BALB/c mice with M. tuberculosis downregulates expression of a Th1-specific costimulatory molecule, M150, on the surface of infected macrophages. The proliferation of Th cells and Th1-cytokine production by these cells are higher in case of M. tuberculosis antigen presentation by uninfected macrophages than by infected macrophages. The difference in inducing interleukin(IL)-2 and interferon (IFN)-gamma secretion is abolished by providing bystander costimulation through M150 on liposomes.

Animals↗
PubMed2000

Gastrointestinal bleeding due to worm infestation, with negative upper gastrointestinal endoscopy findings: impact of enteroscopy.

BACKGROUND AND STUDY AIMS: In cases of hookworm and roundworm infestation, chronic occult bleeding is well known, but acute gastrointestinal bleeding is rarely described. We report five patients with worm infestation who presented with acute massive gastrointestinal bleeding. PATIENTS AND METHODS: Over 1 year, we have encountered 15 patients with obscure gastrointestinal bleeding. All the patients had normal findings on upper gastrointestinal endoscopy. Push enteroscopy was performed in each patient and evaluation of the proximal 40-50 cm of the jejunum was done. RESULTS: Five patients (four men, one woman; average age 50 yr, range 40-60) had worm infestation (two hookworm, three roundworm). All the patients had gastrointestinal bleeding (five had melena, one hematemesis, and two hematochezia) for a duration of 7-14 days. Hemoglobin ranged from 2.8 to 9 g/dl. Push enteroscopy revealed fresh blood in the jejunum, multiple erosions, and hookworms in two patients and roundworms in three patients. Hookworms were retrieved endoscopically in two patients whereas roundworms could be retrieved in only one patient. All the patients were treated with mebendazole (100 mg twice a day for 3 days), iron, and folic acid. Gastrointestinal bleeding subsided in all the patients. CONCLUSIONS: In developing countries, worm infestation should be considered an important cause of obscure acute gastrointestinal bleeding. Evaluation of the jejunum using an enteroscope will result in more frequent diagnosis of worms as a cause of acute gastrointestinal bleeding which might have been classified as obscure gastrointestinal bleeding.

Adult↗
PubMed1999

Development of CD8alpha/alpha and CD8alpha/beta T cells in major histocompatibility complex class I-deficient mice.

Peripheral CD8(+) T cells mainly use CD8alpha/beta, and their development is mainly dependent on the major histocompatibility complex (MHC) class I proteins K(b) and D(b) in H-2(b) mice. In this report, we have shown that the development of CD8alpha/beta TCR-alpha/beta cells in lymphoid organs as well as in intestinal intraepithelial lymphocytes (iIELs) is dependent on the MHC class I K(b) and D(b) proteins. In contrast, TCR-alpha/beta CD8alpha/alpha cells are found mainly in iIELs, and their numbers are unaffected in K(b)D(b) double knockout mice. Most of the TCR-gamma/delta cells in the iIELs also bear CD8alpha/alpha, and they are also unaffected in K(b)D(b) -/- mice. In beta2-microglobulin (beta2m)-deficient mice, all of the TCR-alpha/beta CD8alpha/alpha and CD8alpha/beta T cells disappear, but TCR-gamma/delta cells are unaffected by the absence of beta2m.

Animals↗
PubMed1999

Expression of human gelatinase B in Pichia pastoris.

Full-length human gelatinase B (FLGelB) and its C-terminal truncated form (dGelB) were expressed in Pichia pastoris strain GS115, using the Saccharomyces cerevisiae Mat alpha signal peptide. In both cases, a high level of the secreted protein could be detected by SDS-PAGE. The truncated gene was also expressed using the human gelatinase B native signal peptide. Secretion using the Mat alpha signal peptide was significantly greater than that from the native signal peptide. The recombinant products were purified and characterized biochemically. The recombinant proteins, FLGelB and dGelB, were found to have similar biochemical properties and activity to that of the human gelatinase B native protein.

Base Sequence↗
PubMed1999

Apoptosis of Th1-like cells in experimental tuberculosis (TB).

Th1 cell-induced anti-mycobacterial immunity is lost during a progressive Mycobacterium tuberculosis infection in a susceptible host. This study was designed to test the mechanism of the loss of anti-mycobacterial cell-mediated immune response. We demonstrate that M. tuberculosis infection results in increased Fas expression and decreased Bcl-2 expression in CD4+ T cells. When CD4+ T cells are stimulated in vitro, they show increased apoptosis and decreased production of IL-2 and interferon-gamma (IFN-gamma) but not of IL-4. These changes may result in selective apoptosis of Th1-like cells, leading to the loss of cell-mediated immune response against M. tuberculosis.

Animals↗
PubMed1999

Leishmania donovani infection of a susceptible host results in CD4+ T-cell apoptosis and decreased Th1 cytokine production.

The disease visceral leishmaniasis is caused by a protozoan parasite, Leishmania donovani and is characterized by depressed cell-mediated immunity (CMI) and unhindered parasite growth in a susceptible host. The opposite trend is observed in a resistant host. However, the mechanism of this loss of CMI during the progressive disease is unknown as yet. In this report, we demonstrate that more than 40% of CD4+ T cells from a susceptible host undergo apoptosis resulting in a significant decrease in interleukin (IL)-2 and interferon (IFN)-gamma secretion, leaving IL-4 secretion unaffected. These changes are not apparent in the case of CD4+ T cells derived from a resistant host. The data reported here suggest that experimental Leishmania donovani infection leads to selective deletion of the IL-2 and IFN-gamma-secreting cells but not Th2-like cells in a susceptible but not a resistant host.

Animals↗
PubMed1999

"Talon cusp-heredity origin"--a case report.

Talon cusp is a very unusual anomalous structure of tooth. The etiology is still unknown. It may be due to mal-interaction between ecto and mesoderm of epithelial bulgings present on premaxillary region at the time of complex odontogenesis. Genetics may have some role in the formation of Talon cusp. Talon cusp may cause clinical complications.

Adult↗
PubMed1998

Cloning, expression, purification, and immunocharacterization of placental protein-14.

Human placental protein-14 (PP-14), a member of the lipocalin superfamily, shares homology at the level of the primary and secondary structures with bovine beta-lactoglobulin. It is the most prominent endometrial protein synthesized by the glandular cells of endometrium under estrogen priming and progesterone stimulation. The temporal and spatial expression of PP-14 in the female reproductive tract combined with its biological activities ex vivo suggest that this glycoprotein probably plays an essential physiological role in the regulation of fertilization, implantation, and maintenance of pregnancy. We proposed to elucidate the molecular mechanisms involved in the function of this protein. A prerequisite to such investigations on any protein is the availability of sufficient amounts of the same in a homogenous form. Therefore, recombinant DNA technology was employed. The PP-14 cDNA was obtained from the first-trimester endometrial tissue RNA by RT-PCR using unique primers. After confirming the identity of the gene, the protein was expressed in Escherichia coli and purified to homogeneity. The gene was also cloned and expressed in Pichia pastoris to obtain the protein product in a glycosylated form. The recombinant proteins were immunocharacterized using a cross-reactive antibody raised to bovine beta-lactoglobulin. Polyclonal antiserum raised to the E coli expressed PP-14 also bound to the native PP-14 from amniotic fluid suggesting that recombinant PP-14 may be exploited to elucidate functional aspects of the protein.

Amino Acid Sequence↗
PubMed1998

Production of recombinant human bile salt stimulated lipase and its variant in Pichia pastoris.

hBSSL and its truncated variant hBSSL-C cDNA clones were expressed in Pichia pastoris using two different signal peptides, native signal peptide and invertase signal peptide, respectively, to facilitate secretion of the recombinant proteins into the culture medium. Both recombinant proteins were secreted into the culture medium to a level of 45-50 mg/liter in shake flask cultures. Native signal peptide of hBSSL was recognized in P. pastoris and was cleaved at the same site as in humans. The level of expression of the hBSSL gene was found to be dependent on the number of its copies integrated into the host chromosome. The multicopy transformant clone was found to be very stable. When grown and induced in a fermentor, the level of accumulation of the recombinant hBSSL in the culture medium improved from 50 mg/liter in shake flask cultures to 300 mg/liter. The recombinant hBSSL purified from the culture supernatant was found to be similar to the native hBSSL in its biochemical properties except for the lectin-binding profile.

Chromosomes, Fungal↗
PubMed1998

Leishmania donovani infection of a susceptible host results in apoptosis of Th1-like cells: rescue of anti-leishmanial CMI by providing Th1-specific bystander costimulation.

A protective immune response against Leishmania donovani infection is mediated by T-helper type 1 (Th1) cells. Th1 induced cell-mediated immunity (CMI), as assessed by anti-leishmanial DTH response, is lost in a susceptible host such as BALB/c mice. Although the impaired Th1 function eventuates in unhindered parasite growth and in manifestation of the susceptible phenotype, the mechanism of down-regulation of the Th1 function is yet to be elucidated. Here, we provide evidence that the parasite down-regulates the expression of a Th1-specific costimulatory molecule, M150, on the surface of infected BALB/c mice-derived macrophages. Th cells are rendered unresponsive to anti-CD3 Ab-mediated stimulation after interaction with infected macrophages. The anergized T cells produce much less IL-2, IL-4 and IFN-gamma compared to those T cells which were costimulated using normal macrophages. The defect in proliferation, anti-CD3 Ab induced unresponsiveness and IFN-gamma but not IL-4 production can be restored by providing bystander costimulation through M150. These results not only unfold a novel immune evasion strategy used by the parasite but also clarify the mechanism of Th1 cell debilitation during the disease. Recovery of Th1 cytokine production by bystander costimulation through M150 may help in formulating a new strategy for the elimination of intracellular parasites.

Animals↗
PubMed1997

Selective dysfunction of ventricular electrode-endocardial junction following DC cardioversion in a patient with a dual chamber pacemaker.

A patient with a dual chamber pacemaker received two transthoracic (anterior-anterior paddle position) shocks for VF. Subsequent analysis of stimulation thresholds revealed a marked rise in the ventricular threshold only. Later, she received three additional shocks (anterior-posterior paddle position) during electrophysiological study to terminate episodes of induced VT. Following these, the ventricular stimulation threshold was > 10 V at 2.0 ms. The selective damage at the ventricular site in this patient led us to carry out in vitro studies that revealed a preferential shunting of high electrical energy into the ventricular lead, as compared to the atrial lead, following a DC shock. These observations may explain the selective pacing malfunction observed in out patient.

Burns, Electric↗
PubMed1997

Mycotic keratitis in children: epidemiologic and microbiologic evaluation.

PURPOSE: To evaluate the demographic features, clinical profile, and laboratory diagnosis in cases of mycotic keratitis in children. METHODS: We retrospectively analyzed 211 cases of mycotic keratitis in children younger than 16 years over a 5-year period in a tertiary eye center. Culture-proven cases of fungal keratitis were reviewed. RESULTS: Trauma was the most common predisposing factor (55.3%), followed by associated systemic illness (11.2%), previous ocular surgery (9.8%), and others. Corneal injury contaminated with vegetable matter was responsible for 60.5% of traumatic cases. Aspergillus species were the most frequent isolates (39.5%). Others included Fusarium (10.7%), Alternaria (10.2%), Curvularia (7.4%), and Penicillium (7%). A seasonal variation in the incidence of mycotic keratitis revealed a peak incidence in the months of September and October. One hundred sixty-two children (76.7%) cooperated for examination and scraping under topical anesthesia with or without sedation. General anesthesia for scraping was required in 49 (23%) of 211 children for corneal scraping. Gram stains of corneal scraping were positive for hyphal elements in 54.5% of cases, and potassium hydroxide wet-mount preparation was positive in 90.2% of cases. CONCLUSIONS: This study highlights important risk factors and organisms responsible for mycotic keratitis in children.

Adolescent↗
PubMed1995

Basal promoter elements as a selective determinant of transcriptional activator function.

In eukaryotes, activation of transcription involves an interplay between activators bound to cis-regulatory elements and factors bound to basal elements near the start site of transcription. The basal elements, for example the TATA box or proximal sequence element (PSE) of small nuclear RNA (snRNA) promoters, nucleate the assembly of basal transcription complexes, components of which interact with activators. Although one basal transcription complex can interact with many activators, it is unclear whether different basal transcription complexes can direct different responses to particular activators. We show here that changing the arrangement of basal elements can alter the response to transcriptional activation domains. Indeed, in the human U6 snRNA promoter, point mutation of either a TATA box or PSE results in diametrically opposed responses to VP16- and Sp1-derived activation domains. These basal elements can even discriminate small changes in an activation domain. Thus the arrangement of basal promoter elements provides a mechanism for differential regulation of transcription.

Amino Acid Sequence↗
PubMed1995

Functional characterization of the cis-regulatory elements of the rat ribophorin I gene.

The ribophorin I gene encodes a rough endoplasmic reticulum (RER) specific membrane protein which is a subunit of the oligosaccharyltransferase. To establish the functional activity of its promoter region we have performed transient gene transcription experiments employing plasmid constructs that contain 5' flanking regions of the ribophorin I gene cloned upstream of the CAT reporter gene. Among the restriction fragments obtained from the 1.3-kilobase 5' flanking region, a proximal fragment (-42 to +24) containing two GC-rich elements was required for basic promoter activity, while a fragment (-364 to +24) encoding an additional GC-box and an octamer like motif at -233 conferred the maximal promoter activity. In order to investigate the functionality of an octomer-like sequence co-transfection experiments were performed with Oct-2 cDNA and the CAT reporter gene containing the ribophorin I fragment (-364 to +24). A 3-4-fold increase in the transcriptional activity was observed with this construct. In addition, gel shift experiments showed Oct-2 binding to this construct. These results indicate that Oct-2 is most likely involved in the regulation of the ribophorin I gene transcription. We suggest that the GC-rich elements are necessary for constitutive ribophorin I expression while octamer motif binding proteins function synergistically with the GC-rich element binding proteins to increase the expression of the ribophorin I gene during the proliferation of RER.

Animals↗