Biomedical subjects

A Choudhury

Publications and source records attributed to A Choudhury.

At least 55 records · Page 3Linked to original sources

PubMed2000

Human MUC4 mucin cDNA and its variants in pancreatic carcinoma.

The human MUC4 gene is not expressed in normal pancreas; however, its dysregulation results in high levels of expression in pancreatic tumors. To investigate the tumor-associated expression, MUC4 cDNA was cloned from a human pancreatic tumor cell line cDNA expression library using a polyclonal antibody raised against human deglycosylated mucin and RT-PCR. Pancreatic MUC4 cDNA shows differences in 12 amino acid residues in the non-tandem repeat coding region with no structural rearrangement as compared with tracheal MUC4. The full-length MUC4 cDNA includes a leader sequence, a serine and threonine rich non-tandem repeat region, a central large tandem repeat domain containing 48 bp repetitive units, regions rich in potential N-glycosylation sites, two cysteine-rich domains, EGF-like domains, and a transmembrane domain. We also report the presence of a new EGF-like domain in MUC4 cDNA, located in the cysteine-rich region upstream from the first EGF-like domain. Four distinct splice events were identified in the region downstream of the central tandem repeat domain that generate three new MUC4 cDNA sequences (sv4, sv9, and sv10). The deduced amino acid sequences of two of these variants lack the transmembrane domain. Furthermore, two unique forms of MUC4 (MUC4/Y and MUC4/X) generated as a result of alternative splicing lack the salient feature of mucins, the tandem repeat domain. A high degree of polymorphism in the central tandem repeat region of MUC4 was observed in various pancreatic adenocarcinoma cell lines, with allele sizes ranging from 23.5 to 10.0 kb. MUC4 mRNA expression was higher in differentiated cell lines, with no detectable expression in poorly differentiated pancreatic tumor cell lines.

Alternative Splicing↗

PubMed2000

Redescription of Crepidostomum opeongoensis Caira, 1985 (Trematoda: Allocreadiidae) from fish hosts Hiodon alosoides and Hiodon tergisus (Osteichthyes: Hiodontidae).

Crepidostomum opeongoensis Caira, 1985 (Trematoda: Allocreadiidae) is redescribed from fish hosts Hiodon alosoides and Hiodon tergisus in southern Manitoba. The redescription adds details regarding the surface morphology and reproductive structures of the parasite not described previously. These include the characteristic tegumental papillae around the oral opening, paired papillae on the ventral and dorsal surfaces of the body, and tegumental bosses on the dorsal surface of the forebody. Crepidostomum opeongoensis co-occurred with its hypothesized sister species. Crepidostomum illinoiense, in all host individuals harboring the former, and both species are characteristic of hiodontids. The association of C. opeongoensis with hiodontids and the absence of hiodontids in Lake Opeongo where C. opeongoensis was originally reported indicate that the Algonquin Park region may have had hiodontids in the past and that the life cycle may be completed at present without the original fish host.

Animals↗

PubMed1999

Significance of Cryptosporidium in acute diarrhoea in North-Eastern India.

In a hospital-based study, stool samples from 2095 patients of all ages were examined for different fungal, protozoal and bacterial enteropathogens over a period of 2 years (July 1994-June 1996). Cryptosporidium was detected in 151 specimens (7.2%) and was the third commonest pathogen found. The highest prevalence of this organism was in the group aged 16-45 years and during the rainy months (July-Oct.). Diarrhoea caused by the protozoon was of mild to moderate severity and features of dysentery were absent. Amongst other enteropathogens, Candida albicans was the most frequently isolated, followed by enteropathogenic and enterotoxigenic Escherichia coli, Salmonella spp., Campylobacter jejuni, Entamoeba histolytica, Giardia duodenalis (lamblia), Shigella spp., Vibrio cholerae and Aeromonas spp.

Acute Disease↗

PubMed1998

Coordination of an array of signaling proteins through homo- and heteromeric interactions between PDZ domains and target proteins.

The rapid activation and feedback regulation of many G protein signaling cascades raises the possibility that the critical signaling proteins may be tightly coupled. Previous studies show that the PDZ domain containing protein INAD, which functions in Drosophila vision, coordinates a signaling complex by binding directly to the light-sensitive ion channel, TRP, and to phospholipase C (PLC). The INAD signaling complex also includes rhodopsin, protein kinase C (PKC), and calmodulin, though it is not known whether these proteins bind to INAD. In the current work, we show that rhodopsin, calmodulin, and PKC associate with the signaling complex by direct binding to INAD. We also found that a second ion channel, TRPL, bound to INAD. Thus, most of the proteins involved directly in phototransduction appear to bind to INAD. Furthermore, we found that INAD formed homopolymers and the homomultimerization occurred through two PDZ domains. Thus, we propose that the INAD supramolecular complex is a higher order signaling web consisting of an extended network of INAD molecules through which a G protein-coupled cascade is tethered.

Animals↗

PubMed1998

Human leukemia-derived dendritic cells: ex-vivo development of specific antileukemic cytotoxicity.

The human myeloid leukemias are a diverse group of disorders characterized by massive clonal expansion of myeloid cells showing variable degrees of differentiation block. Leukemic dendritic cells were generated in culture from chronic myelogenous leukemia (CML). These were used to stimulate autologous T cells to develop leukemia-specific cytotoxicity. Available data suggest that the cells responsible for the cytolytic activity are at least in part CD8+ and HLA restricted in their function. Additional data suggest that some anti-CML cellular activity may be Fas mediated. T-cell receptor studies provide evidence for an oligoclonal response implying a recognition of a limited number of antigens. We have used culture techniques similar to those used for CML to study the ability of AML cells to differentiate toward dendritic cells. Four of five patients have shown acute leukemia-derived dendritic cells. This work offers an avenue for the development of novel strategies for the control of human myeloid leukemias.

Cytotoxicity Tests, Immunologic↗

PubMed1998

Lissorchis macropharynx n. sp. (Digenea: Lissorchiidae) from the shorthead redhorse, Moxostoma macrolepidotum (LeSueur) (Osteichthyes: Catostomidae).

Lissorchis macropharynx (Digenea: Lissorchiidae) is described from the shorthead redhorse, Moxostoma macrolepidotum (LeSueur) (Catostomidae) from the Assiniboine River drainage of southern Manitoba. The species is morphologically similar to Lissorchis hypentelii in possessing a similar body shape (widest at the ovarian region and a relatively long hindbody), a distinctly trilobed and relatively posteriorly situated ovary, and in the distribution of the vitellaria. It differs from L. hypentelii and all other lissorchiids in possessing a massive pharynx that is generally equal in length to or longer than the oral sucker and almost as deep as the cervical region of the body it occupies, a protruding acetabular region with an aspinose ridged/folded and grooved anterior surface, and a massive cirrus sac that reaches the level of the ovary. Museum specimens of Lissorchis spp. from M. macrolepidotum from other drainages in Manitoba and Wisconsin were also identified as L. macropharynx.

Animals↗

PubMed1998

Pharmacokinetics and biodistribution of genetically-engineered antibodies.

Monoclonal antibodies (MAbs), because of their inherent specificity, are ideal targeting agents. They can be used to deliver radionuclides, toxins or cytotoxic drugs to a specific tissue or malignant cell populations. Intact immunoglobulin (IgG) molecules have several practical limitations of their pharmacology; their relatively large size of approximately 150,000 daltons leads to a slow clearance from the blood pool and the body resulting in significant exposure to normal organs with limited quantities delivered to tumors. The IgG molecule shows a relatively poor diffusion from the vasculature into and through the tumor. Attempts to modify the pharmacology of the Ig molecule have classically involved the use of proteases to generate F(ab')2 and Fab' fragments with molecular weights of approximately 100,000 and 50,000 daltons, respectively. Fv fragments of IgG are one of the smallest size functional modules of antibodies that retain high affinity binding of an antigen. Their smaller size, approximately 25,000 daltons, enables better tumor penetration and makes them potentially more useful than a whole antibody molecule for clinical applications. Molecular cloning and expression of the variable region genes of IgG has greatly facilitated the generation of engineered antibodies. A single-chain Fv (scFv) recombinant protein, prepared by connecting genes encoding for heavy-chain and light-chain variable regions at the DNA level by an appropriate oligonucleotide linker, clears from the blood at much faster rate than intact IgG. The scFv fragment can retain an antigen-binding affinity similar to that of a monovalent Fab' fragment; this however, represents a relative decrease in binding affinity when compared to intact antibodies. The scFv with its faster clearance and lower affinity results in a lower percent-injected dose localizing in tumors when compared to the divalent IgG molecule. This may be adequate for imaging but probably not for therapy. The valency of the MAb fragment is critical for the functional affinity of an antibody to a cell surface or a polymeric antigen. In attempts to generate multivalent forms of scFv molecules, non-covalently linked scFv dimeric and trimeric molecules, disulfide linked dimeric scFvs, as well as covalently linked chimeric scFvs have been studied. These multivalent scFvs generally have a higher functional affinity than the monovalent form resulting in better in vivo targeting. Another way to alter the pharmacology of the scFvs is to modify its net charge. Charge-modified scFvs with desired isoelectric points (pI), have been prepared by inserting negatively charged amino acids on the template of the variable region genes. This can help to overcome undesirable elevations in renal uptake seen with most antibody fragments. In conclusion, genetic manipulations of the immunoglobulin molecules are effective means of altering stability, functional affinity, pharmacokinetics, and biodistribution of the antibodies required for the generation of the "magic bullet".

Animals↗

PubMed1997

Disruption of T cell tolerance by directing a self antigen to macrophage-specific scavenger receptors.

Breakdown of immune self tolerance is speculated to cause autoimmune diseases, but most studies on tolerance use foreign molecules as targets. In this study, we show another approach using delivery of a maleylated self protein to macrophage-specific scavenger receptors. Mice generate Abs against the maleylated form of a ubiquitous self Ag, mouse serum albumin (MSA), although native MSA is nonimmunogenic. This generation of anti-maleyl MSA Abs depends on binding of maleyl MSA to scavenger receptors in vivo, since coinjection of a serologically unrelated scavenger receptor ligand inhibits it, suggesting that the Ab response is T cell dependent. Spleen cells as well as nylon adherence-purified splenic T cells from maleyl MSA-immune mice proliferate in response to both maleyl MSA and MSA; this response is blocked by anti-MHC class II mAbs, and the autoimmune cells can recognize at least five 15-mer peptides from the MSA sequence, establishing that T cell tolerance to MSA has been broken in these mice. Maleyl MSA and MSA are recognized equally well, provided the scavenger receptor-specific delivery of maleyl MSA is blocked during stimulation in vitro, indicating that maleyl MSA-specific non-self peptides are unlikely to play a major role in the observed disruption of T cell tolerance. Thus, delivery of some self molecules to scavenger receptors may lead to disruption of immune tolerance. These results are relevant to mechanisms of immune tolerance and the etiopathogenesis of autoimmunity.

Animals↗

PubMed1997

Use of leukemic dendritic cells for the generation of antileukemic cellular cytotoxicity against Philadelphia chromosome-positive chronic myelogenous leukemia.

The success of adoptive immunotherapy for the treatment of leukemia depends on the generation of T cells that can specifically react with malignant cells. Dendritic cells (DCs) are important antigen-presenting cells in the development of antileukemic T-cell responses. In this study, we generated DCs from peripheral blood cells of patients with chronic myelogenous leukemia (CML). CML cells incubated concurrently with granulocyte-macrophage colony-stimulating factor, interleukin-4, and tumor necrosis factor-alpha in vitro developed morphologic and phenotypic characteristics of DCs. Fluorescence in situ hybridization showed the presence of t(9;22) in the nuclei of these cells, indicating that they were leukemic in origin. These cells were potent stimulators of lymphocyte proliferation in specific in vitro assays for DC function. Autologous T cells stimulated with in vitro-generated, leukemic DCs displayed vigorous cytotoxic activity against CML cells but low reactivity to major histocompatability complex-matched normal bone marrow cells. Cytotoxic activity against CML targets was fourfold to sixfold higher using DC-stimulated autologous T cells than with autologous T cells expanded by culture with interleukin-2 alone. DC-stimulated T cells also inhibited growth of CML clonogenic precursors in colony-forming assays in vitro. These results suggest that cytokine-driven in vitro differentiation of CML cells results in generation of DCs with potent T-cell stimulatory function. In vitro-generated DCs can be effectively used as antigen-presenting cells for the ex vivo expansion of antileukemic T cells.

Cell Differentiation↗

PubMed1997

Recognition of the parasite infected cell surface determinants by homologous antiserum raised against infected cell membranes.

Identification of neo-antigenic determinant(s) on parasite infected cell surface is important to control intracellular infections. Such determinant(s) on the surface of intact Plasmodium berghei infected erythrocytes have not been conclusively demonstrated. To generate polyclonal antiserum selectively recognizing the parasite infected cell surface determinant(s), in natural state, we have examined the efficacy of the homologous immunizations, in BALB/c mice, with the membrane rich preparation of: i) erythrocytes in vivo infected with Plasmodium berghei and, ii) macrophages in vitro infected with Leishmania donovani. Anti-infected erythrocyte membrane antiserum specifically recognized, albeit at low level, the infected cell surface as determined by flow cytometry and immunoelectron microscopy. Immunoprecipitation of radiolabeled antigens revealed at least three parasite proteins of > 205 kDa, 160 kDa and 100 kDa specifically present on infected erythrocyte surface. Normal uninfected erythrocytes did not react with the antiserum. Anti-L. donovani-infected macrophage membrane antiserum also recognized only infected macrophage surface and not the normal macrophages. Thus, the approach may find wide application in delineating disease specific determinant(s) on the infected cell surface, particularly to those where animal models are available.

Animals↗

PubMed1997

Crepidostomum percopsisi n. sp. (Digenea: Allocreadiidae) from the trout perch (Percopsis omiscomaycus) of Dauphin Lake, Canada.

Crepidostomum percopsisi n. sp. is described from the small intestine of the trout perch (Percopsis omiscomaycus) in Dauphin Lake, Manitoba. It is morphologically similar to Crepidostomum isostomum, which has been reported from the trout perch and several other species of fish. It differs from C. isostomum based on the vitellaria confined to the hindbody of the worm, size and shape of the cirrus, size of the testes, and its greater body length. A comparison of our specimens with those illustrated and identified as C. isostomum from trout perch indicates that such specimens are identical to larger specimens of C. percopsisi recovered by us from trout perch in May. To date, C. percopsisi has only been reported from the trout perch of Dauphin Lake, Lake Winnipeg, and Oneida Lake, which suggests host specificity.

Animals↗

PubMed1996

Lipid and fatty acid composition of muga silkworm, Antheraea assama, host plants in relation to silkworm growth.

The lipid and fatty acid composition of the leaves (tender, medium and mature) of muga host plants, Machilus bombycina, Litsaea monopetala (primary food plants) and L. cubeba and L. salicifolia (family: Lauraceae) (secondary food plants) was investigated by standard procedures, gas chromatography after saponification and esterification. The total lipid content of M. bombycina and L. monopetala leaves was recorded to be higher (16 and 18 g%), respectively, than that of L. cubeba (10 g%) and L. salicifolia (12 g%). GC analysis identified the presence of eight fatty acids (C14 to C22) and the concentration varied from 0.0297 to 8.1572 g% dry leaf powder. Among the fatty acids, (C14 to C22), polyunsaturated fatty acids were recorded to be highest in concentration in mature leaves of the primary host plants. The concentrations of saturated and polyunsatuated fatty acids were found to be at a minimum level in all the types of leaves of secondary muga host plants.

Animals↗

PubMed1996

Character of a wheat amylase inhibitor preparation and effects on fasting human pancreaticobiliary secretions and hormones.

BACKGROUND & AIMS: Amylase inhibition induces carbohydrate tolerance, satiety, and weight loss and prolongs gastric emptying, effects that may be useful in the treatment of obesity and non-insulin-dependent diabetes mellitus. The aim of this study was to determine (1) purity of a wheat amylase inhibitor preparation, (2) intraduodenal concentration of the wheat amylase inhibitor preparation that inhibits > 90% amylase activity (which causes carbohydrate malabsorption), and (3) if the inhibitor alters pancreaticobiliary secretions or intraluminal pH. METHODS: High-performance liquid chromatography followed by electrophoresis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis were used for characterization. Groups of 3 subjects received intraduodenal infusions of 3.0, 4.5, or 6.0 mg/mL of the inhibitor for 90 minutes during the middle of a 270-minute essential amino acid solution infusion (which stimulates 50% maximal pancreatic enzyme secretion). Pancreatic enzyme and bile acid delivery to the duodenum were measured for a 270-minute period. RESULTS: The inhibitor is 96% protein, 59% containing 0.19, 0.28, 0.38, and 0.53 inhibitors. The 0.38 inhibitor has the most antipancreatic alpha-amylase activity. The inhibitor reduced amylase activity in the duodenum dose dependently (r = 0.7; P = 0.04); > 4 mg/mL inhibited > 90% amylase activity but did not affect delivery of other enzymes or bile acids to the duodenum or gastric or duodenal pH. CONCLUSIONS: The preparation has a high protein purity and a high specific activity against alpha-amylase activity and effectively inhibits human pancreatic amylase activity secreted into the duodenum.

Adult↗

PubMed1996

Diclybothrium atriatum n. sp. (Monogenea:Diclybothriidae) from North American acipenserid fishes with observations on Diclybothrium armatum and Diclybothrium hamulatum.

Diclybothrium atriatum n. sp. is described from North American acipenserid fishes, Acipenser fulvescens Rafinesque (lake sturgeon) and Acipenser brevirostrum Le Sueur (shortnose sturgeon). Diclybothrium atriatum is distinguished from other diclybothriids by the presence of a unique preovarial structure that we name the ventral atrial organ. This structure possesses a ventral orifice leading to a deep groove that widens internally as a crypt, bordered by densely staining granular bodies and more deeply situated glandular cells. Previous records of Diclybothrium armatum Leuckart, 1835 examined in this study from North American acipenserids are referable to D. atriatum. Studies on the anatomy of this and other species of Diclybothrium Leuckart, 1835, D. armatum and Diclybothrium hamulatum (Simer, 1929), reveal the presence of paired multichanelled sperm ducts that are shown to be characteristic of the Diclybothriidae Bykhovskii and Gusev, 1950. Paired sperm ducts, while common in Turbellaria with multiple testes, are unusual amongst Monogenea. The endemicity of Diclybothrium atriatum and specificity for A. fulvescens and A. brevirostrum suggests that speciation of this parasite may have occurred following the isolation of Acipenser L. spp. in central and eastern North America.

Animals↗

PubMed1996

Observations on the morphology, systematics, and biogeography of the genus Truttaedacnitis (Nematoda:Cucullanidae).

All species of Truttaedacnitis Petter, 1974, were examined for a phylogenetic analysis. Morphological studies showed that Truttaedacnitis truttae (Fabricius, 1794) from North America possessed a reduced button-shaped or nipple-shaped caudal mucron that is in contrast to the sharp spike-shaped mucron commonly reported in Eurasian specimens. Histological studies showed that the excretory pore in Truttaedacnitis sphaerocephala (Rudolphi, 1809) was situated near the level of the pseudobuccal capsule between the 2 parts of the ventral cephalic ridge. Cucullanus heterodonti Johnston and Mawson, 1943 is transferred to Truttaedacnitis based on the presence of prominent cephalic plates separated by sutures. An hypothesis regarding interrelationships among Truttaedacnitis spp. was developed using phylogenetic systematics. The anterior excretory pore is a synapomorphy supporting the monophyly of Truttaedacnitis clitellarius (Ward and Magath, 1917) and T. sphaerocephala parasitizing sturgeons. The arrangement of cephalic plates indicates that Truttaedacnitis pybusae Anderson, 1992 and T. truttae may form a monophyletic group and further supports the monophyly of T. clitellarius and T. sphaerocephala. Using unordered character states of different cephalic plates produced a cladogram that suggested that species in holarctic/circumboreal families (Acipenseridae, Salmonidae, Petromyzontidae) may be monophyletic. The systematics and biogeography of the various species of Truttaedacnitis and their host specificity indicate that associations in this parasite genus are very ancient and of a relictual nature, determined largely by past continental and oceanic configurations, and a combination of vicariance and dispersal.

Animals↗

PubMed1995

Chloroquine encapsulated in malaria-infected erythrocyte-specific antibody-bearing liposomes effectively controls chloroquine-resistant Plasmodium berghei infections in mice.

The suitability of liposomes as drug carriers in the treatment of drug-resistant rodent malaria was examined after covalently attaching F(ab')2 fragments of a mouse monoclonal antibody (MAb), MAb F10, raised against the host cell membranes isolated from the Plasmodium berghei-infected mouse erythrocytes, to the liposome surface. The antibody-bearing liposomes thus formed specifically recognized the P. berghei-infected mouse erythrocytes under both in vitro and in vivo conditions. No such specific binding of the liposomes with the infected cells was observed when MAb F10 was replaced by another mouse monoclonal antibody, MAb D2. Upon loading with the antimalarial drug chloroquine, the MAb F10-bearing liposomes effectively controlled not only the chloroquine-susceptible but also the chloroquine-resistant P. berghei infections in mice. The chloroquine delivered in these liposomes intravenously at a dosage of 5 mg/kg of body weight per day on days 4 and 6 postinfection completely cured the animals (75 to 90%) of chloroquine-resistant P. berghei infections. These results indicate that selective homing of chloroquine to malaria-infected erythrocytes may help to cure the chloroquine-resistant malarial infections with low doses of chloroquine.

Animals↗

PubMed1995

Function and cell surface phenotype of dendritic cells from rat cornea.

PURPOSE: To isolate dendritic cells (DC) from rat corneas and to examine their functions and surface markers in vitro. METHODS: Cells were isolated enzymatically from dissected rat corneas and cultured for various intervals of time. Dendritic cells were enriched immunomagnetically from corneal cell preparations using monoclonal antibodies against DC surface antigens and tested for functional activity in lymphocyte stimulation assays. In vivo migration of DC was induced by traumatizing the corneal epithelium. Wholemounts of epithelial sheets were stained immunofluorescently with anti-DC antibodies and examined by confocal microscopy. Dendritic cells isolated from traumatized corneas were tested for functional activity. RESULTS: Corneal DC exhibited the properties of other members of the DC family, i.e., low buoyant density, lymphoid DC-specific markers, and lymphostimulatory function. In fresh unfractionated cell preparations of normal cornea, no functional activity was detected. However, DC immunomagnetically purified from fresh preparations were functionally active. Injury to the corneal epithelium induced the migration of DC from the periphery to the central cornea; DC measured in this situation showed significantly increased functional activity. Finally, IL-1 beta and GM-CSF enhanced the functional activity of corneal DC. CONCLUSIONS: Corneal DC have lymphostimulatory capacity in situ, but they may be maintained in a state of latency by the suppressive influence exerted by neighboring cells. Injury to the corneal epithelium results in functional activation of the corneal DC, which may be caused by cytokines such as IL-1 beta or GM-CSF. Thus, corneal DC may be important in the immune regulation of the anterior segment.

Animals↗

PubMed1994

Purification and partial characterization of a heparin-binding lectin from the marine clam Anadara granosa.

The heparin-binding lectin, Anadarin MS, from the plasma of the marine clam Anadara granosa is purified through affinity chromatography on heparin-Sepharose 4B followed by gel filtration on a Sepharose 6B column. The purified lectin is a pentameric protein of native M(r) 300 kDa and is composed of identical subunits of 60 kDa. The pI value of this Ca(2+)-dependent lectin is 6.2. Anadarin MS agglutinates normal rabbit erythrocytes but not that of human. Aspartic acid, glutamic acid, histidine and glycine are the predominant amino acids. Unlike other reported heparin-binding lectins, Anadarin MS exhibits a unique and strict specificity for iduronic acid containing glycosaminoglycans. This lectin agglutinates infective promastigotes of Leishmania donovani exclusively and can therefore be used as a novel biochemical surface marker for this parasite.

Agglutination↗