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Biomedical subjects

C Vincent

Publications and source records attributed to C Vincent.

At least 235 records · Page 13Linked to original sources
PubMed1988

Biophysical chemical aspects of cellular cryobehavior.

Freezing tolerance and resistance in nature are among the most important and challenging aspects of biochemical adaptation to extreme environments. Some biochemical strategies are known but their mechanism is still poorly understood. Cryopreservation of cells and tissues of sensitive organisms is still generally based on physical chemistry rather than on biophysical chemical mechanisms. This paper describes the main aspects of these problems and features new trends in their study.

Adaptation, Physiological↗
PubMed1987

Rapid binding of beta 2-microglobulin to renal brush-border membranes.

125I-labelled human beta 2-microglobulin binding to rat renal brush-border membranes was assessed by an in vitro assay under near physiological incubation conditions (i.e. low content of albumin). Binding rate was 55 pmol/min per mg protein in the presence of 200 nM of beta 2-microglobulin and degradation rate was negligible versus binding rate. The binding rate was in reasonable agreement with the in vivo reabsorption rate, supporting the hypothesis of proteins binding to the luminal membrane during the process of reabsorption. Mild solubilizing treatment (Triton 0.1%) of brush border after beta 2-microglobulin binding yielded the labelled molecule associated with a high-molecular-weight component. Aminopeptidase activity and binding ability were to a certain extent co-purified during the course of the brush-border preparation, suggesting that most of the beta 2-microglobulin binding sites were localized in the brush-border membranes.

Aminopeptidases↗
PubMed1987

Purification of alpha 1-microglobulin produced by human hepatoma cell lines. Biochemical characterization and comparison with alpha 1-microglobulin synthesized by human hepatocytes.

alpha 1-Microglobulin (alpha 1m) was determined by radio-immunoassay in the supernatants of five human hepatoma cell lines. High amounts of alpha 1m were produced by PLC/PRF/5, intermediate ones by Hep G2 and Hep 3B and very low ones by Malhavu and SK Hepl. alpha 1m isolated from hepatoma cell lines PLC/PRF/5 or Hep G2 supernatants displayed the same physicochemical properties as that purified from human urines: the apparent molecular mass was 26 kDa and the pI from 5.6 to 6.4 as measured after two-dimensional polyacrylamide gel electrophoresis in denaturating conditions; for the native molecule the pI was estimated to be 4.0-4.9. Both urinary and hepatoma alpha 1m migrate as a diffuse band in the alpha zone in agarose gel at pH 8.6 in non-denaturing conditions and present a brown chromophore covalently associated with the molecule. After biosynthetic labelling with [35S]methionine, proteins extracted from hepatoma cell line PLC/PRF/5 and from isolated hepatocytes of human liver were separated by two-dimensional PAGE and transferred to a nitrocellulose membrane. alpha 1m was identified and found to be identical in both cases. However, when compared with the alpha 1m isolated from cell supernatants, less charge heterogeneity but also minor additional spots of higher molecular mass were observed.

Alpha-Globulins↗
PubMed1987

Transfer of frozen-thawed embryos in sheep.

Embryos collected from ewes six days after oestrus were frozen in straws using ethylene-glycol as a cryoprotectant. The efficiency of the simplified freezing and thawing procedure was assessed after transfer, which resulted in an overall survival rate of 58.3 per cent. Forty-two lambs were born from 72 frozen embryos which had been transferred without any attempt to evaluate them after the thawing and sucrose dilution process.

Animals↗
PubMed1987

Human serum pancreatic lipase and trypsin 1 in aging: enzymatic and immunoenzymatic assays.

Two major human pancreatic proteins, lipase and trypsin I, were measured in human sera from 35 elderly healthy adults and 51 young healthy adults. Lipase enzymatic activity was determined by a turbidimetric assay in the presence of colipase; lipase immunoreactive protein and trypsin immunoreactive protein were measured by using immunoenzymatic assays. Serum levels of the pancreatic enzymes were similar in young and elderly adults, with no significant differences between the groups for any of the assays. There was a close correlation between lipase enzyme activity and immunoreactivity in all participants, suggesting that the catalytic activity of lipase was not affected by aging. There were also significant correlations between the levels of immunoreactive lipase protein and immunoreactive trypsin protein, within and between the two groups, suggesting an age-independent relationship between these two pancreatic enzymes.

Adult↗
PubMed1987

Radiation stimulation during the early stationary growth phase in Synechococcus lividus and its correlation with photooxidative stress occurring before the stationary phase.

The effects of chronic gamma radiation at dose rates ranging from 0.058 mGy d-1 on growth rate calculated during the early stationary phase were studied. A stimulatory effect occurred for all doses and for all phases of the cells selected for use in the inoculation of the medium. During the same period, the rate of nucleic acid synthesis was increased in irradiated cultures compared to control cultures. The stimulating effect always occurred in cultures irradiated from the inoculation to the eighteenth day only. This result led us to conclude that the stimulation mechanism depended upon the events occurring at the end of the exponential phase in the deceleration period. Studies on cell metabolism showed that cells presented features of photooxidative stress in this period. Increases in superoxide dismutase, glutathione reductase and glucose-6-phosphate dehydrogenase were observed in irradiated cultures. It was assumed that irradiation at very low doses could help cells to better defend against photooxidative stress by increasing oxidants that activate the glucose metabolism and C5-sugars production and nucleic acid synthesis.

Cyanobacteria↗
PubMed1987

Natural IgM and IgG autoantibodies to epidermal keratins in normal human sera. I: ELISA-titration, immunofluorescence study.

This paper presents a study of autoantibodies (autoAB) to keratins and to epidermis by a double approach associating a specific immunoenzymatic technique and immunofluorescence. The existence of natural autoAB to keratins in all normal human sera was asserted and the heterogeneity of natural autoAB to the epidermis explored. By a sensitive enzyme-linked immunosorbent assay we detected natural IgM and IgG autoAB to keratin polypeptides extracted from human plantar stratum corneum (SC) in 60 randomly selected normal human sera. The interindividual variation factors of their titers were about 100X in IgM and 50X in IgG antikeratin (AK) autoAB. The IgM and IgG AK autoAB titers varied independently. By a semiquantitative indirect immunofluorescence assay we detected in these sera IgM and IgG autoAB that labeled normal epidermis according to various morphologic patterns. The IgG autoAB labeled SC and suprabasal layers (SBL) in 57.4% of sera, SC in 20.4% and SBL in 7.4%. The IgM autoAB labeled SC and SBL in 52% of sera, SC in 24%, SC and SBL plus basal layer (BL) in 18%, and SBL in 2%. Like the titers, the patterns of IgM and IgG autoAB to epidermis were found to be unrelated. The IgG AK autoAB titers were found to significantly correlate only with the IgG autoAB directed to SC + SBL; the IgM AK autoAB titers only with the IgM autoAB directed to SC + SBL + BL. This showed that these patterns of labeling are typical for AK autoAB and that autoAB to SC, which could not be related to AK autoAB, exist in some normal sera. Antikeratin and antiepidermis IgM autoAB titers were found to be strongly correlated to total amounts of IgM assayed by radial immunodiffusion, indicating that the synthesis of these natural IgM autoAB vary in the same way as that of general IgM synthesis. For the AK and antiepidermis IgG autoAB, however, the same correlation to total serum IgG was found to be much weaker.

Adult↗
PubMed1987

Expression of class I-MHC antigens by cultured human epidermis and epidermal allografts.

In vitro grown class II-MHC antigen free epidermal sheets were used as epidermal allografts (EAG) across a major histocompatibility barrier in 20 non-immunosuppressed recipients suffering from leg ulcers. Class I antigens were expressed on cell membranes of basal cell layer only on the epidermal sheets. After grafting, patchy areas of membrane fluorescence were observed among cells from the suprabasal layers on the epidermis from skin biopsies taken between days 5 and 28. All cells of the basal and the suprabasal layers expressed class I antigens on biopsies taken after day 28, as on normal human epidermis. This work demonstrates that class I antigens are expressed by epidermal cells in cultures used for grafting. The absence of rejection cannot be explained by the absence of class I-MHC antigens in EAG.

Adult↗
PubMed1987

B-cell maturation stages of Burkitt's lymphoma cell lines according to Epstein-Barr virus status and type of chromosome translocation.

This study addressed the possible relationship between B-cell maturation stage of Burkitt's lymphoma (BL) cell lines and Epstein-Barr virus (EBV) status, ethnic group, or type of chromosome translocation. Fifty-seven cell lines obtained at the International Agency for Research on Cancer from 51 patients were studied. Cytogenetic analyses of 54 cells lines were available. Cell size, surface immunoglobulins (sIgs), cytoplasmic immunoglobulins (cIgs), mouse red blood cell receptors, and reactivity with various monoclonal antibodies were assessed. Immunoglobulin (Ig) class secretions were measured in the supernatant of 2- and 5-day cultures from 33 cell lines, with the use of a sensitive enzyme-linked immunosorbent assay technique. From this study, BL appears to cover a broad range of the B-cell differentiation sequence, since the following Ig phenotypes were observed: null cells (sIg-, cIg-), large pre-B-cells (intracytoplasmic mu-chains), small B-cells (sIg+, cIg-), and various types of secreting B-cells (sIg+, cIg+). Among the latter, various patterns of cIg could be defined (perinuclear, paranuclear, and vesicular). B-cell maturation stages were correlated with the amount of secreted Ig. In sIg+ cell lines, different classes of Ig were found: 35 IgM, 10 IgM plus IgD, 4 IgG, and 1 IgA. None of the different monoclonal antibodies used was specific to a precise stage of maturation. The stages of maturation were correlated with neither the type of chromosome translocations of BL nor the presence of EBV genome, but the most immature cell lines were all EBV positive and most of them originated from African patients. In contrast with acute lymphoblastic leukemia, the common acute lymphocytic leukemia antigen (CD10) was expressed on nearly all BL cell lines of intermediate maturation stages but only on half of the pre-B ones. In addition, none of the cell lines tested was found to react with CD5 antibodies, which recognize most of the chronic lymphocytic leukemia of the same stage of maturation as that in the B-lymphocyte lineage.

Adolescent↗
PubMed1986

Effects of a very low dose rate of chronic ionizing radiation on the division potential of human embryonic lung fibroblasts in vitro.

Among the various parameters that are supposed to play a role in aging at the cellular level, the "free radical theory" involves biochemical modifications that can be induced by radiation. Human embryonic lung fibroblasts were serially subcultivated at low density under chronic low dose rate irradiation (40 mrad/day) and in a normal environment. Irradiation increases cell attachment and the population doubling/day throughout their entire in vitro lifespan. Consequently, the doubling potential reached by irradiated cells was higher than that of control cultures. Finally, the total number of cells produced under chronic irradiation was 8-14 times higher than in a normal environment. These results are discussed with respect to the increased enzymatic activities (superoxidismutase, catalase, glutathion-reductase, G6PD) found in some irradiated organisms.

Cell Adhesion↗
PubMed1986

The M2 autoantigen of central nervous system myelin, a glycoprotein present in oligodendrocyte membrane.

Autoantibodies with in-vitro demyelinating capacity induced in Hartley and strain 13 guinea pigs with homologous central nervous system (CNS) tissue were used to characterize the target autoantigen M2. Using the Dot Immunobinding technique, M2 was found to be a component of CNS myelin different from basic protein (BP) and from cerebroside. The expression of M2 on oligodendrocytes, cells known to produce CNS myelin, also confirmed that M2 was a component of CNS myelin. Furthermore, the autoradiography of immunoprecipitates formed with radiolabelled guinea pig myelin and analysed in sodium dodecyl sulphate gels showed that M2 was specific to CNS myelin and absent in peripheral nervous system (PNS) myelin. On electrophoresis M2 appeared as two CNS myelin protein bands at the 27 and 54 KD molecular weight levels, distinct from the major protein bands of proteolipid and BP. M2 bands were of glycoprotein nature, as was demonstrated by affinity chromatography of CNS myelin on wheat germ agglutinin (WGA)-Sepharose. A monoclonal antibody induced by BP-free CNS glycoproteins recognized the same bands as anti-M2 serum in guinea pig CNS myelin. This would imply that both M2 bands share common determinants. M2 bands similar to the above in guinea pig were also shown in rat, rabbit and bovine CNS myelin with guinea pig antibodies. The same type of anti-M2 antibodies were induced in rabbit immunized with homologous CNS tissue. Although only a minor component of myelin, M2 is strongly immunogenic compared to BP. M2 antigen could thus be the target of chronic demyelinating processes such as experimental allergic encephalomyelitis.

Animals↗
PubMed1986

[Anti-stratum corneum antibody in the rat esophagus, anti-epidermal keratin and anti-epidermis autoantibodies in rheumatoid polyarthritis and other rheumatic diseases. Diagnostic value and basic aspects].

Eight immunological parameters were explored in 257 patients with various rheumatic diseases including 107 rheumatoid arthritis (RA). IgG and IgM antibodies (AB) reactive with the Stratum Corneum (SC) of rat oesophagus or with the SC of human skin were assayed by 1/2 quantitative indirect immunofluorescence, IgG and IgM auto AB to epidermal keratins by a specific ELISA, and total serum G and M immunoglobulins by radial immunodiffusion. At the threshold we chose (99 p. cent specificity), IgG anti SC of rat oesophagus, were found in 50 of 107 (46.7 p. cent) RA patients: 52.5 p. cent in sero + and 39.6 p. cent in sero - ones. The other parameters, separately considered, had no diagnosis value. In Paget disease serum IgM and IgG and all the AB of M isotype were found to be broken down. In all the groups, the isotype AB were found strongly correlated to each other and to serum IgM. The RA sera with specific AB to the rat oesophagus SC, also labeled human skin SC but these labeling were unrelated to the anti-keratins auto AB level. On the contrary, the anti SC AB of G isotype, detectable on human skin in psoriatic rheumatism did not label rat oesophagus. These results confirm the diagnosis value of IgG AB to SC of rat oesophagus, usually called anti-keratins, who appear as a marker for RA. This work shows the relevance of associating specific immunochemical techniques to immunofluorescence, in order to unravel the antigenic complexity of tissular substrata.

Animals↗
PubMed1985

Differential measurement by ELISA of free and IgA bound alpha1-microglobulin in human serum without prior fractionation.

Two solid phase enzyme immunoassays for the measurement of alpha1-microglobulin (alpha1m) in biological fluids are reported. These assays are complementary and allow the separate determination of free alpha1m and alpha1m bound to IgA without fractionation of the fluids. Specificity, reproducibility and sensitivity of the assays are reported. Differential measurements on normal or pathological sera were compared to the measurements obtained by chromatographic separation of free and bound alpha1m before assay. Results obtained with the enzyme immunoassays were significantly correlated with those obtained by radioimmunoassay.

Alpha-Globulins↗