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Biomedical subjects

A K Sarkar

Publications and source records attributed to A K Sarkar.

At least 19 recordsLinked to original sources

Characterization of aerosols from biomass burning--a case study from Mizoram (Northeast), India.

Physical and optical properties of biomass burning aerosols in Northeastern region, India analyzed based on measurements made during February 2002. Large spatial extent of Northeastern Region moist tropical to moist sub-tropical forests in India have high frequency of burning in annual dry seasons. Characterization of resultant trace gases and aerosols from biomass burning is important for the atmospheric radiative process. Aerosol optical depth (AOD) observed to be high during burning period compared to pre- and post-burning days. Peak period of biomass burning is highly correlated with measured AOD and total columnar water vapor. Size distribution of aerosols showed bimodal size distribution during burning day and unimodal size distribution during pre- and post-burning days. Size distribution retrievals from biomass burning aerosols show dominance of accumulation mode particles. Weighted mean radius is high (0.22 microm) during burning period. Columnar content of aerosols observed to be high during burning period in addition to the drastic reduction of visibility. During the burning day Anderson sampler measurements showed dominance of accumulation mode particles. The diurnal averaged values of surface shortwave aerosol radiative forcing af biomass burning aerosols varies from -59 to -87 Wm(-2) on different days. Measured and modeled solar irradiances are also discussed in the paper.

Air Pollutants↗

Variation in the origin of the inferior alveolar nerve.

The inferior alveolar nerve and maxillary artery were studied in 40 human heads through infratemporal dissection; in one specimen, a unusual variation in the origin of the inferior alveolar nerve and its relationship with the surrounding structures was observed. The inferior alveolar nerve originated from the mandibular nerve by two roots and the second part of the maxillary artery was incorporated between them. An embryologic origin of this variation and its clinical implications is discussed. Because the maxillary artery ran between the two roots of the inferior alveolar nerve, and the nerve was fixed between the foramen ovale and mandibular foramen, tension and compression of the nerve from arterial pulsation could cause mandibular neuralgia.

Adult↗

Synthesis and glycan priming activity of acetylated disaccharides.

Five disaccharides related in structure to the glycans of vertebrate mucins have been chemically synthesized using orthogonal blocking, coupling and deblocking techniques. These include 2-naphthylmethyl 3,4,6-tetra-O-acetyl-beta-D-galactopyranosyl-( 1 --> 4)-2-acetamido-3,6-di-O-acetyl-2-deoxy-beta-D-glucopyranoside (6), 2-naphthylmethyl 2-aceta-mido-3,4,6-tri-O-acetyl-2-deoxy-beta-D-glucopyranosyl-(1 --> 3)-2,4,6-tri-O-acetyl-beta-D-galactopyranoside (14), 2-naph-thylmethyl2,3,4,6-tetra-O-acetyl-beta-D-galactopyranosyl-(1 --> 3)-2-acetamido-4,6-di- O-acetyl-2-deoxy-alpha-D-galactopyranoside (20), 2-naphthylmethyl 2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-beta-D-glucopyranosyl-(1 --> 3)-2-acetamido-4,6-di-O-acetyl-2-deoxy-alpha-D-galactopyranoside (23) and 2-naphthylmethyl 2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-beta-D-glu-copyranosyl-(1 --> 6)-2-acetamido-3,4-di-O-acetyl-2-deoxy-alpha-D-galactopyranoside (27). These per-O-acetylated compounds were fed to U-937 cells to test their ability to prime oligosaccharide synthesis, inhibit glycoprotein biosynthesis and alter adhesion to E-selectin expressed on endothelial cells. The results show that 6, 14, and 20 served as substrates for oligosaccharide synthesis. The generation of glycoside-primed glycans altered the formation of glycoproteins on the cell surface and inhibited cell adhesion dependent on E-selectin.

Acetylation↗

Lysis of MYCN-amplified neuroblastoma cells by MYCN peptide-specific cytotoxic T lymphocytes.

The effectiveness of cell-mediated immunotherapy for cancer can be limited by loss-of-antigen mutations that occur during tumor growth. In neuroblastoma, amplification of the MYCN oncogene correlates with rapid tumor progression and a poor prognosis overall. We propose that the MYCN protein, the high-level expression of which is required for maintenance of the malignant phenotype, would be an ideal target for vaccine therapy. The MYCN-derived S9K peptide (amino acids 7-15; STMPGMICK), which contains an HLA-A1 binding motif, was used to generate CTLs from the peripheral blood lymphocytes of an HLA-A1+ healthy donor and an HLA-A1+ patient with MYCN-amplified neuroblastoma These CTL lines specifically lysed HLA-matched, MYCN-amplified neuroblastoma tumor cells. They did not lyse either HLA-mismatched, MYCN-amplified, or matched/nonmatched, non-MYCN-amplified tumor cells. The CTL activity was inhibited by a monoclonal antibody to a class I HLA monomorphic determinant but not by one specific for HLA class II, consistent with a class I-restricted mechanism of cytotoxicity. Antibodies to CD8, but not those to CD4, also inhibited CTL activity, identifying CD8+ lymphocytes as the effector cell population. These results show that MYCN-derived peptides can serve as tumor-specific antigens and suggest a rational approach to cell-mediated immunotherapy for MYCN-amplified neuroblastoma.

Adult↗

Anatomy of the cystic artery arising from the gastroduodenal artery and its choledochal branch--a case report.

Variations in the branching pattern of the common hepatic artery often occur and may be encountered during cholecystectomy. Variants of the cystic artery, its branches and relations with the biliary structures and blood vessels emphasise the importance of arterial dissection in biliary surgery. In this study, a rare variant of the cystic artery and its choledochal branch is described. The cystic artery arose from the gastroduodenal artery, passed anterior to structures in the free margin of lesser omentum and travelled a long distance before supplying the gall bladder. A long choledochal branch was noted accompanying the common bile duct. Surgical implications of this variation of the cystic and choledochal arteries are discussed.

Arteries↗

Formation of HNK-1 determinants and the glycosaminoglycan tetrasaccharide linkage region by UDP-GlcUA:Galactose beta1, 3-glucuronosyltransferases.

While expression-cloning enzymes involved in heparan sulfate biosynthesis, we isolated a cDNA that encodes a protein 65% identical to the UDP-GlcUA:glycoprotein beta1, 3-glucuronosyltransferase (GlcUAT-P) involved in forming HNK-1 carbohydrate epitopes (3OSO3GlcUAbeta1,3Gal-) on glycoproteins. The cDNA contains an open reading frame coding for a protein of 335 amino acids with a predicted type II transmembrane protein orientation. Cotransfection of the cDNA with HNK-1 3-O-sulfotransferase produced HNK-1 carbohydrate epitopes in Chinese hamster ovary (CHO) cells and COS-7 cells. In vitro, a soluble recombinant form of the enzyme transferred GlcUA in beta-linkage to Galbeta1,3/4GlcNAcbeta-O-naphthalenemethanol, which resembles the core oligosaccharide on which the HNK-1 epitope is assembled. However, the enzyme greatly preferred Galbeta1, 3Galbeta-O-naphthalenemethanol, a disaccharide component found in the linkage region tetrasaccharide in chondroitin sulfate and heparan sulfate. During the course of this study, a human cDNA clone was described that was thought to encode UDP-GlcUA:Galbeta1,3Gal-R glucuronosyltransferase (GlcUAT-I), involved in the formation of the linkage region of glycosaminoglycans (Kitagawa, H., Tone, Y., Tamura, J., Neumann, K. W., Ogawa, T., Oka, S., Kawasaki, T., and Sugahara, K. (1998) J. Biol. Chem. 273, 6615-6618). The deduced amino acid sequences of the CHO and human cDNAs are 95% identical, suggesting that they are in fact homologues of the same gene. Transfection of a CHO cell mutant defective in GlcUAT-I with the hamster cDNA restored glycosaminoglycan assembly in vivo, confirming its identity. Interestingly, transfection of the mutant with GlcUAT-P also restored glycosaminoglycan synthesis. Thus, both GlcUAT-P and GlcUAT-I have overlapping substrate specificities. However, the expression of the two genes was entirely different, with GlcUAT-I expressed in all tissues tested and GlcUAT-P expressed only in brain. These findings suggest that, in neural tissues, GlcUAT-P may participate in both HNK-1 and glycosaminoglycan production.

Amino Acid Sequence↗

Risk assessment of cardiovascular diseases among bank employees--a biochemical approach.

In different epidemiological studies, an association between sedentary life style and incidence of cardiovascular diseases has been demonstrated. This study was carried out among 50 sedentary bank employees working at different nationalized banks within Calcutta City, India and 50 individuals with non-sedentary jobs matched for age group and sex for the control group. Body mass index (BMI), blood pressure (BP), plasma level of glucose, total cholesterol, low density lipoprotein (LDL), very low density lipoprotein (VLDL) and triglycerides (Tgs) were found to be significantly higher whereas high density lipoprotein (HDL) and reduced glutathione were found to be significantly lower among the bank employees as compared with the control subjects. Further, significant alteration in the activity of different enzymes (e.g., glutathione reductase, glutathione S-transferase, glucose 6 phosphate dehydrogenase, catalase and superoxide dismutase) involved in glutathione-mediated antioxidant scavenging systems and increases in lipid peroxidation were also observed in the bank employees. From the study, it appears that the relative risk for cardiovascular disease is increased among bank employees due to the sedentary nature of their jobs.

Adult↗

A study of the effects of pulsed electromagnetic field therapy with respect to serological grouping in rheumatoid arthritis.

The positive role of pulsed electromagnetic field (PEMF) therapy in rheumatoid arthritis (RA) is known. The differential role of serological status of patients in RA is also well known. This paper presents a study of the differential effects of PEMF therapy on the two serological groups of patients. The responses of the seropositive patients are found to be more subdued. Varying effects of the therapy in alleviating the different symptomatologies indicate that the rheumatoid factor (RF) is more resistant to PEMF.

Adolescent↗

Fucosylation of disaccharide precursors of sialyl LewisX inhibit selectin-mediated cell adhesion.

We showed previously that HL-60 and F9 mouse embryonal carcinoma cells will take up and deblock peracetylated Galbeta1-4GlcNAcbeta-O-naphthalenemethanol (Galbeta1-4GlcNAc-NM) and use the disaccharide as a primer of oligosaccharide chains (Sarkar, A. K., Fritz, T. A., Taylor, W. H., and Esko, J. D. (1995) Proc. Natl. Acad. Sci. U. S. A. 92, 3323-3327). We now report that another disaccharide, acetylated GlcNAcbeta1-3Gal-naphthalenemethanol (GlcNAcbeta1-3Gal-NM), has even greater potency and that both compounds will inhibit sialyl LewisX (sLex)-dependent cell adhesion. When fed to U937 cells, acetylated forms of Galbeta1-4GlcNAc-NM and GlcNAcbeta1-3Gal-NM primed oligosaccharides in a dose-dependent manner. Analysis of compounds assembled on Galbeta1-4GlcNAc-NM showed only one product, namely Galbeta1-4(Fucalpha1-3)GlcNAc-NM. In contrast, GlcNAcbeta1-3Gal-NM generated Galbeta1-4GlcNAcbeta1-3Gal-NM, Galbeta1-4(Fucalpha1-3)GlcNAcbeta1-3Gal-NM, NeuAcalpha2-3Galbeta1-4GlcNAcbeta1-3Gal-NM, and NeuAcalpha2-3Galbeta1-4(Fucalpha1-3)GlcNAcbeta1- 3Gal-NM. Both compounds decreased the incorporation of [3H]fucose into cellular glycoconjugates, without affecting the incorporation of [3H]mannosamine, a precursor of sialic acid residues. Moreover, the overall extent of sialylation was not affected based on the reactivity of cells to fluorescein isothiocyanate-conjugated Maackia amurensis lectin. Priming inhibited expression of sLex on cell surface glycoconjugates, which reduced E-selectin-dependent cell adhesion to tumor necrosis factor-alpha-activated human umbilical vein endothelial cells. GlcNAcbeta1-3Gal-NM and Galbeta1-4GlcNAc-NM represent starting points for making enzyme-specific, site-directed inhibitors of glycosyltransferases that could act in living cells.

Acetylation↗

Role of detritus in a general prey-predator model of Sunderban Estuary, India.

In this paper we consider a general deterministic model consisting of a micro-organism pool living on detritus of mangrove litter and its invertebrate predators. Here the growth rate of micro-organism is assumed in the general form and Holling type-II functional response of the invertebrate predators is incorporated. We find a globally attracting steady state for some parameter values, and a stable limit cycle for some other parameter values. It is also shown that there exists Hopf-Andronov bifurcation with respect to the control parameters. It is found that the specific growth rate and the food conversion efficiency rate of invertebrate predator governs the dynamics of such a model.

Animals↗

Unusual beta-D-xylosides that prime glycosaminoglycans in animal cells.

The biosynthesis of glycosaminoglycans (GAG) takes place while the polysaccharide chains are usually attached to a proteoglycan core protein. Cells also will assemble GAG chains on beta-D-xylosides containing hydrophobic aglycones. In order to evaluate the relationship of the structure of the sugar to priming activity of the glycoside, we synthesized beta-D-xyloside analogs in which the hydroxyls were substituted with hydrogen, fluorine, -O-methyl, amino, -O-isopropyl, and -O-benzyl groups. Epimers at the 2-, 3-, and 4-position of xylose also were made. Their ability to prime GAGs was tested in Chinese hamster ovary cells by measuring 35SO4 incorporation into polysaccharide chains and by assaying the transfer of galactose to the xylosides by galactosyltransferase I (UDP-D-galactose:xylose beta1-4-galactosyltransferase) in vitro. All of the analogs failed to act as primers of GAGs in vivo and as substrates in vitro with the following exceptions. Substitution of 2-OH and 3-OH with -OCH3 were active at high concentration (1 mM), but the deoxygenated derivatives were inactive. Efficient priming also occurred on a derivative with fluorine instead of the 3-OH group, suggesting that the oxygen atoms at C-2 and C-3 were involved as hydrogen bond acceptors. Methylated and deoxy analogs at C-4 were inactive, due to the loss of the acceptor hydroxyl group. Interestingly, benzyl-beta-D-threo-pentopyranos-4-uloside (4-keto derivative) and benzyl-4-methyl-beta-D-xyloside, with a methyl group in place of an axial hydrogen at C-4, primed GAG chains. Priming by these unusual xylosides suggests the possibility of designing inhibitors of GAG synthesis based on xyloside analogs with reactive groups in key positions.

Animals↗

Hyperuricemia and pregnancy induced hypertension--reappraisal.

Serum uric acid estimation was done in forty primigravidae with pregnancy induced hypertension and twenty normotensive primigravida in the third trimester of pregnancy, at delivery and six weeks postpartum. The mean serum uric acid levels in normotensive women in the antenatal period and at delivery were 4.65 +/- 0.33 and 4.88 +/- 0.23 mg% and in mild PIH were 5.42 +/- 0.55, 6.14 +/- 0.76 mg%, respectively. Level of serum uric acid in mild PIH was significantly higher than normotensive women (P). In severe PIH, the mean serum uric acid levels were 6.65 +/- 0.60, 8.24 +/- 1.09 mg% in antepartum and at delivery respectively which was significantly more than control group and mild PIH group women (P). However, no differences was observed, in the serum uric levels between these groups during the postpartum period. Serum uric acid level of 5.5 mg or more was observed to be an indicator of PIH. Levels of serum uric acid did show a high positive correlation with the severity of PIH in relation to hypertension and proteinuria. Hyper uricemia (more than 5.5 mg% is associated with increased perinatal morbidity and mortality.

Female↗

Synthesis and glycosaminoglycan priming activity of three disaccharides related to the linkage region tetrasaccharide of proteoglycans.

To test if disaccharides might serve as primers of oligosaccharide synthesis in animal cells, we synthesized 2-naphthyl O-(beta-D-galactopyranosyl)-(1 --> 4)-beta-D-xylopyranoside, 2-naphthyl O-(beta-D-galactopyranosyl)-(1 --> 3)-beta-D-galactopyranoside, and 2-naphthyl O-(beta-D-glucopyranosyluronic acid)-(1 --> 3)-beta-D-galactopyranoside. These three disaccharides are related to subunits of the linkage tetrasaccharide of heparan sulfate and chondroitin sulfate chains in animal cell proteoglycans. The disaccharides were synthesized with coupling efficiencies of 40-70% using thioglycosides or by activating the monosaccharides with trichloroacetimidate. The structures of these compounds were confirmed by 1H NMR, 13C NMR and elemental analysis. The ability of these disaccharides to prime glycosaminoglycan chains was examined in a Chinese hamster ovary cell mutant, p gsA 745, which lacks xylosyltransferase. The missing enzyme renders the cells dependent on exogenous primers for making glycosaminoglycan chains. 2-Naphthyl O-(beta-D-galactopyranosyl)-(1 --> 3)-beta-D-galactopyranoside and 2-naphthyl O-(beta-D-glucopyranosyluronic acid)-(1 --> 3)-beta-D-galactopyranoside did not stimulate glycosaminoglycan synthesis, but 2-naphthyl O-(beta-D-galactopyranosyl)-(1 --> 4)-beta-D-xylopyranoside at high concentration primed chains. The peracetylated derivative (2-naphthyl O-(2,3,4,6-tetra-O-acetyl-beta-D-galactopyranosyl)-(1 --> 4)- 2,3-di-O-acetyl-beta-D-xylopyranoside) primed chains at lower concentration (100 microM), suggesting that cells took up the compound and removed the acetyl groups apparently in the compartment where glycosaminoglycan synthesis occurs.

Acetylation↗

Disaccharide uptake and priming in animal cells: inhibition of sialyl Lewis X by acetylated Gal beta 1-->4GlcNAc beta-O-naphthalenemethanol.

Inhibitors of glycosylation provide a tool for studying the biology of glycoconjugates. One class of inhibitors consists of glycosides that block glycoconjugate synthesis by acting as primers of free oligosaccharide chains. A typical primer contains one sugar linked to a hydrophobic aglycone. In this report, we describe a way to use disaccharides as primers. Chinese hamster ovary cells readily take up glycosides containing a pentose linked to naphthol, but they take up hexosides less efficiently and disaccharides not at all. Linking phenanthrol to a hexose improves its uptake dramatically but has no effect on disaccharides. To circumvent this problem, analogs of Xyl beta 1-->6Gal beta-O-2-naphthol were tested as primers of glycosaminoglycan chains. The unmodified disaccharide did not prime, but methylated derivatives had activity in the order Xyl beta 1-->6Gal(Me)3-beta-O-2-naphthol > Xyl beta 1-->6Gal (Me)2 beta-O-2-naphthol >> Xyl beta 1-->6Gal(Me)beta-O-2-naphthol. Acetylated Xyl beta 1-->6Gal beta-O-2-naphthol also primed glycosaminoglycans efficiently, suggesting that the terminal xylose residue was exposed by removing the acetyl groups. The general utility of using acetyl groups to create disaccharide primers was shown by the priming of oligosaccharides on peracetylated Gal beta 1-->4GlcNAc beta-O-naphthalenemethanol. This disaccharide inhibited sialyl Lewis X expression on HL-60 cells.

Animals↗

Studies on in vivo induction of cytotoxic T lymphocyte responses by synthetic peptides from E6 and E7 oncoproteins of human papillomavirus type 16.

Induction of cytotoxic T lymphocyte (CTL) responses is an important defense mechanism against infectious agents, specifically viruses. In the present investigation we employed a mouse assay system we previously developed, for rapid induction of CTLs by synthetic peptides from E6 and E7 oncoproteins of human papillomavirus type 16 (HPV-16). In particular, we compared the efficiency of CTL induction by HPV-16 peptides synthesized as linear monomers with those containing a dipalmitoyl-lysine-glycine-glycine (P2-KGG) moiety at the amino-terminus. Our results identified a 15-amino-acid peptide from E6(Q15L, aa 43-57) to be capable of inducing CTLs in vivo and addition of the lipid tail significantly increased CTL induction over that seen with the linear form of the peptide. Further, we identified a shorter peptide, V1OC, with 9 of 10 amino acids overlapping with Q15L peptide (aa 49-58) to be capable of inducing CTLs against both V1OC and Q15L. In case of E7 protein, our results demonstrated usefulness of P2-KGG moiety for enhanced CTL induction by previously identified CTL epitope peptides Q19D (aa 44-62) and R9F (aa 49-57). CTLs induced by both the E6 and E7 peptides were MHC class I-restricted and exhibited strict allele specificity and CD8+ phenotype. Our results showing enhanced cell-mediated immune responses with lipid-tailed forms of peptides add strength to the concept of a synthetic peptide-based vaccine for prophylaxis and therapy of HPV-associated cervical cancer.

Amino Acid Sequence↗

Biosynthesis of heparan sulfate on beta-D-xylosides depends on aglycone structure.

We have reported that 3-estradiol-beta-D-xyloside primes heparan sulfate synthesis in Chinese hamster ovary cells and that the proportion of heparan sulfate made rises with increasing concentration of xyloside (Lugemwa, F.N. and Esko, J.D. (1991) J. Biol. Chem. 266, 6674-6677). Using estradiol as a guide, we varied the structure of the aglycone and showed that beta-D-xylosides containing two fused aromatic rings efficiently prime heparan sulfate. Thus, 2-naphthol-beta-D-xyloside primed heparan sulfate at low dose (< or = 10 microM) and the proportion of heparan sulfate increased with concentration (up to 50% of total glycosaminoglycan). Various ring additions and heterocyclic ring substitutions altered the efficiency of heparan sulfate priming, but had no effect on the overall level of glycosaminoglycan synthesis. Replacement of the bridging oxygen with sulfur (2-naphthalenethiol-beta-D-xyloside) increased the efficiency of heparan sulfate priming. Priming of heparan sulfate correlated with hydrophobicity of the xyloside, but several exceptions suggested that the chemical structure of the aglycone played an equally important role. Interestingly, the heparan sulfate chains generated on 2-naphthol-beta-D-xyloside showed a 2-fold decrease in the proportion of disaccharides containing 6-O-sulfate groups and a striking diminution in non-sulfated iduronic acid containing disaccharides compared to the chains attached to cellular proteoglycans. Thus, both the type of glycosaminoglycan made on a xyloside and its fine structure depends on the aglycone.

Animals↗