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Biomedical subjects

D N Misra

Publications and source records attributed to D N Misra.

At least 37 records · Page 2Linked to original sources

Adsorption of 4-methacryloxyethyl trimellitate anhydride (4-META) on hydroxyapatite and its role in composite bonding.

The adsorption of 4-methacryloxyethyl trimellitate anhydride (4-META) was studied from ethanol and dichloromethane onto synthetic hydroxyapatite (containing about 1.5 monolayers of physisorbed water) in order to study its role in restorative composite bonding to teeth. The adsorption isotherm of 4-META was S-shaped and reversible from ethanol, and followed the Langmuir plot at lower concentrations. The isotherm was irreversible from dichloromethane, and a constant amount of adsorbate was removed from the solutions above a certain concentration. The irreversibly adsorbed compound was completely removed by washing with ethanol. Therefore, the bonding between teeth and the restorative resin containing 4-META as a coupling agent is micromechanical and not chemical in nature. An analysis of isotherms showed that the benzene rings of the adsorbate molecules lie flat on the surface for both solvents. The molecules adsorbed from ethanol rotate about the solvent-modified (esterified) or unmodified carboxylic anhydride moieties with methacrylate hydrocarbon groups which are folded upward. However, the molecules adsorbed from dichloromethane remain fixed to the surface without rotation, and their methacryloxyethyl groups are folded over the rings. The diametral tensile strength of a BIS-GMA polymer with adsorbate-covered apatite was approximately equal to that of the composite filled with untreated apatite.

Acrylates↗

Isolation and characterization of rat trophoblast cells.

Previous immunohistochemical studies of the rat placenta using specific alloantisera and/or monoclonal antibodies showed that the basal zone trophoblasts stained for Pa and Aa class I major histocompatibility complex (MHC) antigens and for the human SP1-related antigen. In an effort to isolate the basal zone trophoblast cells from the rat placenta, we used these markers to assess the degree of purification of the cells separated by density gradient centrifugation using either Ficoll-Hypaque or Percoll as the gradient medium. The cells were put either on the top or at the bottom of discontinuous density gradients in the range of 1.005-1.10 g/ml. The cell separation profiles for the two media were different. With Percoll, most of the trophoblast cells (80-95%) were collected at the density gradients 1.04/1.06 and 1.06/1.08, whereas with Ficoll-Hypaque, these gradients separated only a small fraction (4-23%) of the trophoblast cells, and most of them pelleted at the bottom of the tube. The trophoblast cells separated by Ficoll-Hypaque, however, showed fewer contaminant cells than those separated by the Percoll gradients.

Animals↗

Differential expression of MHC class I antigens on the placenta of the rat. A mechanism for the survival of the fetal allograft.

In some mating combinations in rats, there is a maternal antibody response to the maternal antigenic components of the placenta without any previous immunization of the mother. The highest response occurs in the WF (u) female mated to the DA (a) male, and it is against a unique MHC-encoded class I antigen, the Pa antigen, and not against the major allele-specific transplantation antigen of the DA strain, RT1.Aa. The development of mAbs to the Pa and Aa antigens allowed us to localize these antigens on the placenta and to explore the reason for the differential antibody response to them using immunohistochemical and biochemical techniques. Both antibodies reacted with the WF X DA placenta and stained the endovascular and interstitial trophoblast of the decidua, the basal trophoblast, Reichert's membrane, and the yolk sac epithelium, but they did not stain the labyrinthine trophoblast. Blocking studies showed that each antibody reacted with a separate molecule in the placenta. Anti-class II mAbs reactive with the a or u haplotype did not stain the WF X DA, DA X DA, or WF X WF placenta; hence, there are no class II antigens in the placenta. Electron microscopic studies of the semiallogeneic WF X DA placenta using the immunogold technique with both single- and double-labeling showed that only the Pa antigen was expressed on the surface of the basal trophoblast, but that both the Pa and Aa antigens were in the cytoplasm of these cells; neither antigen was found in the labyrinthine trophoblast. By contrast, the placenta from the syngeneic DA X DA mating expressed both the Pa and Aa antigens on the surface of the basal trophoblast as well as in the cytoplasm; neither antigen was found in the labyrinthine trophoblast. These observations were quantified morphometrically using electron photomicrographs of single-labeled tissues. Both the Pa and Aa antigens isolated from the plasma membrane of lymphocytes have heavy chains of 46 kD, but those antigens isolated from the plasma membrane of basal trophoblast cells have heavy chains of 43 kD. Based on densitometric measurements of autoradiographs, the Pa/Aa ratio in the basal trophoblast membrane is 23.5, whereas it is 0.46 in lymphocyte membranes. These studies show that there is differential regulation of the expression of class I antigens on basal trophoblast cells in semiallogeneic pregnancies, but not in syngeneic pregnancies, such that the major allele-specific transplantation antigen is scarcely expressed on the surface of the basal trophoblast.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Adsorption of N-phenylglycine on hydroxyapatite: role in the bonding procedure of a restorative resin to dentin.

The adsorption of N-phenylglycine (NPG) onto synthetic hydroxyapatite from ethanol solutions was studied to elucidate the role of surface-active moieties in determining the character of adsorption, and to explore the role of NPG in a bonding procedure of restorative resin to dentin. The adsorption isotherm of N-phenylglycine is reversible (and Langmuirian) from ethanol (99.8%). At maximum adsorption the phenyl rings of the adsorbed molecules lie flat on the surface and the carboxylate oxygens and amino nitrogens are anchored to the surface. The N-phenylglycine serves as an amine accelerator since the peroxide containing monomer polymerizes with the adsorbate-covered apatite. The diametral tensile strength of this composite is approximately equal to the composite filled with untreated apatite. The role of various factors contributing to adhesive strength in the bonding procedure is also discussed.

Adsorption↗

Carbohydrate moieties of rat MHC class I antigens.

The rat major histocompatibility complex class I antigens RT1.Au and RT1.Eu from the u haplotype and RT1. An from the n haplotype were labeled with 14C-asparagine or with 3H-fucose, mannose, galactose, and N-acetylglucosamine. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed complete removal of radioactivity from the sugar-labeled antigen heavy chains by digestion with glycopeptidase F, an enzyme that removes N-linked glycans completely. High performance liquid chromatography analysis of the tryptic digests of the mixed sugar-labeled and asparagine-labeled antigens demonstrated that all the sugar-labeled peptides were coincident with asparagine-labeled peptides. The An antigen showed three glycopeptides, each of which had different amounts of sugar radioactivity. The antigens Au and Eu showed two glycopeptides with different amounts of radioactivity but at identical positions in the two antigens. Antigen Eu had an additional glycopeptide with a lower amount of radioactivity. The positions of the glycopeptides from the Au and Eu antigens were different from those of the An antigen. The peptide profiles of the 14C-asparagine-labeled Au and Eu antigens demonstrated distinct differences between the molecules. The results of this study show that: (a) all the glycans on rat class I antigens are N-linked, as they are on H-2 and HLA class I antigens; (b) there are compositional differences among the glycans in each of the three antigens; (c) the glycosylation pattern of the rat class I antigens is similar to that of the mouse class I antigens, which contain two or three glycans, in contrast to that of the human class I antigens, which contain only one glycan; and (d) the antigens Au and Eu from the same haplotype are more closely related to each other than they are to the An antigen.

Animals↗

Comparison of rat MHC class I antigens by peptide mapping.

Monoclonal antibodies specific for the rat major histocompatibility complex (MHC) class I antigens RT1.An, RT1.Au, and RT1.Eu were used for immunoprecipitation of antigens biosynthetically radiolabeled with 14C- or 3H-labeled arginine, lysine, and tyrosine; with arginine or tyrosine alone; and with or without tunicamycin in the culture medium. Heavy chains of the glycosylated and unglycosylated antigens were purified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and their tryptic and chymotryptic peptides were compared by high performance liquid chromatography. The antigens coded by the same locus in two different haplotypes (An and Au) differed by 30%, whereas the products of two different loci in the same haplotype (Au and Eu) differed only by 1-3%. Comparative analysis of the data for samples labeled with single amino acids indicated that two amino acids in Au have been substituted by an arginine and probably by a tyrosine residue, respectively, in Eu. The high degree of homology between the products of the A and E loci in the same haplotype accounts for the difficulty in detecting recombinational events within the MHC of the rat by classical serological approaches.

Amino Acid Sequence↗

The major histocompatibility complex of the rat.

Immunogenetic studies in the rat began with the investigation of blood group antigens (1-7) and evolved into the investigation of histocompatibility antigens and transplantation phenomena (8-17). In the course of this work, several nomenclature systems evolved that were eventually reconciled in a series of comparison studies (18-24). The biennial Workshops on Alloantigenic Systems in the Rat held under the aegis of the Transplantation Society (24) have provided the forum for the continued evolution of this field. In addition, several reviews (24-31) and books (32-35) have provided periodic summaries of different aspects of rat immunogenetics. This review will focus on the structure of the major histocompatibility complex (MHC) of the rat from the serological, biochemical, and molecular points of view.

Amino Acid Sequence↗

Water on apatites.

Adsorption of water was studied gravimetrically at 23 degrees C in an open system at several relative humidities on a variety of apatitic calcium phosphates including enamel, deproteinized enamel, and bone mineral. The amount of adsorbed water increases linearly with the surface areas of the synthetic apatites and does not appear very sensitive to calcium to phosphorus ratio of the apatites. The adsorption results correlate very well up to about two monolayers with a conventionally determined isotherm. Higher uptake of water even by "deproteinized" enamel or bone may be due to the presence of pore structure and incompletely removed organic matter.

Adsorption↗

Complexes of iron cations with N-phenylglycinate or oxalic acid.

N-phenyglycine plays a very important role in obtaining adhesion of restorative composites to dentin and enamel (Bowen et al., 1982a). In a systematic investigation, ferric or ferrous N-phenylglycinate complexes formed when aqueous ferric nitrate or ferrous chloride was combined with solutions of potassium N-phenylglycinate in stoichiometric proportions. The molal ratios of iron ion to N-phenylglycinate ion in each complex were confirmed by osmolality measurements with a freezing-point osmometer. The reaction of aqueous solutions of oxalic acid with ferric nitrate indicated formation of ferric oxalate complexes with a stoichiometry of Fe2(oxalate)3 in solution, using Job's method of continuous variations (1925;1928).

Adhesiveness↗

Adsorption of benzoic acid on pure and cupric ion-modified hydroxyapatite: implications for design of a coupling agent to dental polymer composites.

The adsorption isotherms of benzoic acid on synthetic hydroxyapatite (containing about 1.5 monolayers of physisorbed water) were studied from ethanol, dimethylsulfoxide, p-dioxane, methylene chloride, and benzene to discern the role of solvent in the process. The adsorption is reversible from the first three solvents and follows the Langmuir plots. It is irreversible from the last two, and a constant amount of absorbent is removed from solutions above a certain concentration. The isotherms of potassium benzoate on the apatite from ethanol and dimethyl sulfoxide were reversible. The isotherms of the acid on cupric ion-modified apatite surfaces from ethanol and benzene were identical with those obtained on the pure hydroxyapatite. This may demonstrate that any "surface chelation" with the cation may not be a significant factor for adsorption to occur. The adsorptive behavior seems to depend upon the interplay of hydrogen-bonding among the solute, the solvent, and the hydrated apatite surface. The capability of a solvent to hydrogen-bond may determine whether adsorption from it will be reversible or irreversible. Based upon its compatibility with a solvent, the benzene ring is upright or lies flat on the surface. The adsorbed molecules rotate about the center of the carboxylate groups which are hydrogen-bonded to the surface. These factors should be considered in designing or selecting a suitable surface-active moiety for a coupling agent between tooth mineral and a restorative resin.

Adsorption↗

Production of monoclonal antibodies against RT2 antigens of the rat and the definition of a new locus, RT9, linked to RT2.

Efforts were made to generate hybridomas producing monoclonal antibodies against the RT2a, RT2b and RT3a antigens of the rat. While a number of hybridomas from each of five different fusions was found to produce antibodies against the RT2b antigen, no hybridoma producing antibody to the RT2a antigen could be detected among those generated in six different fusions. No stable hybridoma secreting antibody specific for the RT3a antigen could be established in three different fusions, although one positive culture was detected. In the course of this work, a monoclonal antibody against a new antigenic specificity was detected in one of the two strain combinations in which the anti-RT2b antibodies were raised. The locus controlling this antigen, which was designated RT9, maps 4.9 (2.2-12.6) cM from RT2.

Alleles↗

Adsorption of zirconyl salts and their acids on hydroxyapatite: use of the salts as coupling agents to dental polymer composites.

Zirconyl methacrylate (I) and zirconyl-2-ethylhexanoate (II) were synthesized, and their adsorption isotherms from solutions onto synthetic hydroxyapatite were studied. The isotherms of methacrylic and 2-ethylhexanoic acids were also determined from the same solvents. The adsorption of I was irreversible from methylene chloride, and that of II was irreversible from cyclohexane. The adsorption in both cases was constant from solutions above a certain concentration, and exhaustive below this threshold concentration. Both compounds rendered the dried apatite powder extremely hydrophobic; however, the adsorbate was slowly washed off by excess water. The configuration of the adsorbate molecules, deduced from the maximum adsorption and other adsorption characteristics of the two compounds, indicated that: (i) in both cases the adsorbate may be held to the surface by concerted hydrogen bonding of the carboxylate and zirconyl oxygen atoms; and (ii) the hydrocarbon moieties in both adsorbates expose themselves toward the solution, thereby making the dried surface hydrophobic. The adsorptive behavior of the respective acids was similar to that of the salts. Polymer, filled with synthetic hydroxyapatite covered with irreversibly adsorbed I, had a diametral tensile strength about 50% greater than that of the polymer filled with untreated apatite. The strength of the composite was not affected by treatment of the apatite with II or with the acids.

Acids↗

Toilet facilities in a large Indian teaching hospital.

While assessing the total medical care provided by any large hospital in a community, hardly any stress is laid on the sanitary facilities which the hospital provides and which form such an integral part of every hospital patient. We studied the toilet facilities of M.L.B. Medical College, Jhansi. They seem to be adequate for the ambulant patient but are insufficient for the bedridden and for those who have weak legs. The findings are discussed.

Activities of Daily Living↗

Analysis of class I MHC antigens in the rat by monoclonal antibodies.

Monoclonal antibodies (mAb) were made against class I MHC antigens of the i (mAb 42,70,39) and u (mAb 68-D) haplotypes in the rat by using specific strain combinations in order to obtain reagents for identifying the products of the RT1.An, RT1.Au, and RT1.Eu loci. These antibodies were hemagglutinating only; were IgG except for mAb 68-D3, which had a defective heavy chain; reacted identically with MHC-congenic strains and with their inbred donor strains; and precipitated class I MHC antigens. Strain distribution, sequential immunoprecipitation, and peptide mapping studies were used to define the specificities of the mAb, and the assignments were checked by comparing the specificities of the mAb with those of haplotype-specific alloantisera. The specificities were the following: mAb 42, An; mAb 68-D, Au; mAb 70, Eu; and mAb 39, an antigen encoded by a locus different from A and E. This new locus was designated RT1.F, and the allele detected by mAb 39, as Fa. The serologic data place RT1.F between RT1.A and RT1.D. The plasma membranes of DA.1I(BI) lymphocytes contain comparable amounts of An, Eu, and Fa antigens but express them on the cell surface in the order An much greater than Eu greater than Fa.

Animals↗