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Biomedical subjects

M A Fletcher

Publications and source records attributed to M A Fletcher.

At least 127 records · Page 7Linked to original sources
PubMed1986

After discharge from the intensive care nursery: what then for the neonatologist?

The field of neonatology is expanding in many aspects. Not only is care of the neonatal patient evolving, but there now is also the need for follow-up long after the neonatal period. The neonatologist has ever increasing roles in preparing patients for discharge, communicating with all relevant follow-up care givers, updating them with advances in the field of neonatology as it relates to follow-up care, and coordinating whatever medical and developmental follow-up is necessary to guarantee that all the effort spent during an ICN stay will not be wasted by unaddressed, potentially preventable problems arising after discharge.

Humans↗
PubMed1985

Mothers of infants with the acquired immunodeficiency syndrome. Evidence for both symptomatic and asymptomatic carriers.

Sixteen mothers of 22 infants with the acquired immunodeficiency syndrome (AIDS) or AIDS-related complex were followed up for evidence of clinical and immunologic abnormalities. With one exception, all mothers were clinically well at delivery but had evidence of immune dysfunction, with T-cell abnormalities and inverted T4/T8 ratios and/or elevation of serum immunoglobulin levels, particularly of IgG. During a follow-up period that averaged 30 months, AIDS developed in five of the mothers and AIDS-related complex in seven. Twelve subsequent pregnancies in 11 mothers produced four affected infants, suggesting that mothers can be persistently infected. Six mothers were delivered of subsequent infants who remain unaffected. These results suggest that the mothers are the likely source of infection in non-transfusion-associated cases of AIDS or AIDS-related complex in infants, that mothers have persistent immunologic abnormalities, and that they are at increased risk of developing AIDS or AIDS-related complex.

Acquired Immunodeficiency Syndrome↗
PubMed1985

Decreased natural cell-mediated cytotoxicity per effector cell in acquired immunodeficiency syndrome.

Natural cell-mediated cytotoxicity (CYT) was determined using whole blood samples from 32 patients with the acquired immunodeficiency syndrome (AIDS) and 12 with AIDS-related complexes (ARC) and correlated with the number of putative natural killer cells (NK) bearing surface marker Leu 11a. Mean percent cytotoxicity (%CYT) was significantly lower in AIDS and ARC patients compared to that of normal controls. Mean %NK cells in AIDS patients was significantly higher than in normals, while %NK cells in ARC patients did not differ significantly from that of normals. In contrast, the absolute number of NK (NNK) cells in AIDS patients did not differ significantly from that of normals, while the NNK cells in ARC patients was significantly lower than those in normals. When the data were expressed in kinetic lytic units per NK cell (KLU/NK = maximum number of K562 cells killed per NK cell in 4 hours), significantly lower KLU/NK were observed in both AIDS and ARC patients compared to that in normals. Our data show that NK cells in AIDS and ARC patients are in a less active state compared to those in normals.

Acquired Immunodeficiency Syndrome↗
PubMed1984

Values for some serum analytes in high-risk infants, two days postpartum.

Using a continuous-flow (Technicon SMAC) analyzer, we measured the concentration of 20 analytes in capillary blood sera from 149 neonates in an intensive-care unit. Included were term and pre-term babies, both groups with and without asphyxia. The values obtained, especially for those enzymes and analytes that are related to renal function, offer baseline data against which the effect of therapeutic and physiological changes can be evaluated.

Apgar Score↗
PubMed1983

Immunochemical studies of infectious mononucleosis--XI. comparison of heterophile antibody inhibitors from the erythrocyte membranes of four mammalian species.

Immunochemical comparisons were made of the reactivity of membrane glycoproteins from horse, bovine, sheep and goat erythrocytes with heterophile antibodies of infectious mononucleosis. The four receptors were tested as competitive inhibitors of a sandwich-type solid-phase radioimmunoassay and of agglutination of glycoprotein-latex reagents by infectious mononucleosis serum. The results of this study showed that the bovine glycoprotein had a superior reactivity with this heterophile antibody system and sheep erythrocyte glycoprotein was the least reactive. The latter had negligible ability to displace 125I-bovine glycoprotein and was a very poor inhibitor of the agglutination of a bovine glycoprotein-latex reagent by infectious mononucleosis serum. Horse and goat glycoproteins were more efficient inhibitors than sheep glycoprotein but less active than the preparation from bovine red cells. All of the inhibitory activity of sheep, horse and goat glycoproteins, and a major portion of that of the bovine glycoprotein was destroyed by neuraminidase treatment. We have termed this receptor--shared by all four species--the Paul-Bunnell receptor, since by definition Paul-Bunnell antibody is a sheep erythrocyte agglutinin which is also reactive with horse, bovine and goat erythrocytes. The neuraminidase (and alkaline borohydride) resistant receptor of bovine glycoprotein has been designated the Bo receptor because it is not common to the other three species.

Agglutination Tests↗
PubMed1983

Serodiagnosis of infectious mononucleosis with a bovine erythrocyte glycoprotein.

A glycoprotein from bovine erythrocyte membrane was evaluated in two immunoassays as a reagent for the serodiagnosis of infectious mononucleosis (IM). We previously reported that a partially purified preparation of this glycoprotein, when attached to latex beads, agglutinated in the presence of IM heterophile antibody. In the present study, we used a highly purified form of the glycoprotein both as an agglutinating reagent, covalently bound to latex, and in a solid-phase sandwich-type radioimmunoassay (RIA) for IM antibody detection in a larger population of patients. We tested serum samples from college students with symptoms suggestive of IM with the latex reagent (143 samples) and with the RIA (245 samples). Correlation of these two tests, both with each other and with the classical differentially absorbed, agglutination tests for Paul-Bunnell antibody in IM sera, using fresh sheep or horse cells, was excellent (greater than 97% agreement). The new tests also corresponded in most cases with a rapid, unabsorbed preserved horse erythrocyte slide test. However, in this study of 245 samples, both apparent false-positives (5 samples) and apparent false-negatives (3 samples) were observed with this slide test. In conclusion, we found that the bovine glycoprotein as a reagent can facilitate the diagnosis of IM, giving results comparable to those with erythrocyte agglutination tests on differentially absorbed sera. The advantages are ease and speed of performance (latex test), potential for automation (RIA test), stability and uniformity of the glycoprotein reagent (latex and RIA tests), and most importantly, the ability to use unabsorbed sera (latex and RIA tests).

Animals↗
PubMed1982

Biochemical and morphological properties of bovine erythrocyte membrane glycoproteins.

The major and minor sialoglycoproteins of the bovine erythrocyte have been solubilized and extensively purified. A comparison of composition revealed that the major glycoprotein had 77% carbohydrate and 23% peptide, and the minor one had 27% carbohydrate and 73% peptide. Molar ratios of sugars were related, however, the major glycoprotein had twice as much galactose and sialic acid as did the minor glycoprotein. Molecular weights, estimated from retardation coefficients of mobility in sodium dodecyl sulfate gel electrophoresis, were 55,000 for the major glycoprotein and 34,000 for the minor glycoprotein. The glycoproteins were studied by electron microscopy before and after delipidation and after ultracentrifugation. The major glycoprotein, prior to delipidation, formed large micelles. After delipidation, the major glycoprotein could not be visualized suggesting that it did not form aggregates in aqueous solution. The minor glycoprotein was visualized as rather uniform spherical aggregates (62 A average diameter) which tended to form short chains and small clumps. These characteristic aggregates were seen both before and after delipidation. After ultracentrifugation, fixation and sectioning both glycoproteins appeared to have formed microcrystalline arrays with average periodicity of 49 A.

Animals↗
PubMed1982

Immunochemical studies of infectious mononucleosis--X. Characterization of a glycoprotein from horse erythrocytes which reacts with Paul-Bunnell antibody.

A highly purified preparation of horse erythrocyte glycoprotein was prepared from an aqueous ethanolic extract of hemoglobin-free membranes. The subunit apparent mol. wt was 30,000. In aqueous solution the glycoprotein formed globular aggregates of 93 +/- 16 A diameter. The glycoprotein had a receptor for the Paul-Bunnell antibody of infectious mononucleosis which was associated with an O-glycosidically linked oligosaccharide and dependent on the presence of N-glycolylneuraminic acid. In addition the glycoprotein had a neuraminidase-sensitive receptor for human peripheral blood lymphocytes. Fifty per cent inhibition of the rosetting of sheep red cells by 4 x 10(5) lymphocytes was caused by 30 microgram of glycoprotein.

Amino Acids↗
PubMed1982

Purified membrane fractions from mammary tumor cells elicit biological reactivity in in vitro cell-mediated immune reactions.

Lithium chloride (LiCl) aggregation followed by centrifugation through a sucrose step gradient was used to obtain purified plasma membranes from two sublines of mouse mammary adenocarcinoma. These two tumor lines were chemically induced by treatment of a hyperplastic nodule with 7,12-dimethylbenzanthracene (DMBA). One line, D1-DMBA-3, has been found to be immunogenic to the host of origin, while the other, D1-DMBA-2, does not elicit specific tumor immunity as previously tested in in vivo and in vitro immune reactions. The purified membrane fractions were assayed for protein, DNA and sialic acid content as well as enzymatic markers of membrane purity. When tested in blastogenesis and cytotoxicity reactions, membrane-containing fractions from the D1-DMBA-3 immunogenic tumor were found to be stimulatory to spleen cells of D1-DMBA-3 tumor bearers over a ten-fold protein concentration. Spleen cells from normal mice do not respond in these reactions to the various tumor fractions. No reactivity was observed when non-membrane-containing preparations were used as stimuli in the cell-mediated immune reactions. The specificity of these reactivities was further demonstrated by the lack of responses when fractions from the non-immunogenic D1-DMBA-2 tumor were tested in parallel in our in vitro assays. The data presented indicate that the procedure employed is useful for the isolation of membrane-associated, tumor-specific antigens which can be easily quantitated and still retain biological activity in in vitro tests of cell-mediated immunity.

Animals↗
PubMed1982

Horse erythrocyte glycoprotein-latex reagent that reacts with infectious mononucleosis heterophile antibody.

A sialoglycoprotein from horse erythrocytes was isolated in essentially homogeneous form and found to contain the neuraminidase-sensitive determinant of the horse erythrocyte for Paul-Bunnell heterophile antibodies of infectious mononucleosis. This reactivity was retained after covalent coupling of the antigen to latex particles. The latex reagent has greater stability (greater than 3 years) than either fresh or preserved horse erythrocytes. It can be used in a direct slide test; no absorption of the serum is necessary. The new test compared favorably with some standard tests for infectious mononucleosis antibody which use horse erythrocytes. Agreement of the latex test with the absorbed fresh horse cell test was 100%, and agreement with a stabilized horse erythrocyte spot test was 94%. The latex test also agreed 100% with a radioimmunoassay for the detection of heterophile antibody with 125I-labeled horse erythrocyte glycoprotein. The new latex test was compared with a bovine erythrocyte glycoprotein-latex test, and a correlation of 94% was observed. In addition, it was compared with the Wöllner test (enzyme-treated sheep cell-absorbed sheep cell agglutination test) with which it agreed in 92% of samples tested.

Animals↗
PubMed1982

Immunochemical studies of infectious mononucleosis. IX. Heterophile antigen associated with a glycoprotein from the bovine erythrocyte membrane.

A glycoprotein was isolated from the membrane of the bovine erythrocyte by refluxing the acetone- and ethanol-extracted stroma residue with 75% ethanol. The glycoprotein was purified by phosphocellulose chromatography, ethanol precipitation, lipidsolvent extraction and DEAE chromatography. The glycoprotein appeared to have two serological determinants, both reactive with antibodies present in the sera of patients with infectious mononucleosis. One of the determinants is similar to the Paul-Bunnell heterophile antigen found on sheep erythrocytes. It is dependent on carbohydrate, including sialic acid residues. Another specificity, seemingly not shared by sheep erythrocytes to any great extent, is resistance to neuraminidase and to alkaline borohydride treatment and thus it may be located either on the polypeptide portion of the molecule or on an alkali-stable oligosaccharide. The purified glycoprotein comprises 73% amino acids. Carbohydrate components and their molar ratios were sialic acid (1.0): galactose (1.5): N-acetylglucosamine (1.1): N-acetylgalactosamine (0.5): mannose (0.1).

Amino Acids↗
PubMed1982

Immunochemical studies of infectious mononucleosis. VIII. A glycoprotein from sheep erythrocytes with sialic acid-dependent receptor properties.

A glycoprotein was solubilized from sheep erythrocyte membranes with hot aqueous ethanol. The glycoprotein was purified by phosphocellulose chromatography, ethanol precipitation, lipid solvent extraction, and DEAE chromatography. In water solution the glycoprotein existed as globular aggregates with a diameter of 7.1 +/- 2.2 nm. In the presence of sodium dodecyl sulfate, 80% of the material exhibited a subunit m.w.app of 27,000. Approximately 10% of the material had a m.w.app of only 9000 and another 10% had a m.w.app of 35,000. All three fractions were reactive with Paul-Bunnell heterophile antibody from the sera of patients with infectious mononucleosis and with Limulus polyphemus lectin. These activities were destroyed by neuraminidase treatment. Complete inhibition of the rosetting of sheep red blood cells by 4 X 10(5) human peripheral blood lymphocytes was seen at 100 to 200 micrograms glycoprotein/ml. Neuraminidase-treated glycoprotein was not inhibitory. Pronase-derived sialoglycopeptide was inhibitory. Most likely, the receptor for lymphocytes resides in the carbohydrate portion of the glycoprotein. By using 125I-glycoprotein, binding studies were carried out that yielded an estimate of approximately 2 X 10(5) binding sites for sheep erythrocyte glycoprotein per lymphocyte. Purified glycoprotein contained 44.4% amino acid. Carbohydrate components and their molar ratios were sialic acid (1.0): galactose (1.0):N-acetylglucosamine (1.3): N-acetylgalactosamine (1.2).

Animals↗
PubMed1982

Two-dimensional gel electrophoresis of membrane proteins from the R3327 prostate adenocarcinoma.

The Dunning rat prostate adenocarcinoma (R3327) is a reliable model that shares many similarities with the human tumor. Two sublines of the tumor, G and H, represent opposite extremes in histology and growth rate. Purified membrane fractions from G and H solid tumors were isolated by sucrose gradient. Tumor and normal prostate membrane proteins were labeled with 125I, incubated with G and H antisera, and precipitated by adsorption of antibody-antigen complexes to staphylococcal Protein A. Proteins were resolubilized and electrophoresed on two-dimensional gels, and the gels were autoradiographed. A total of eight labeled proteins were precipitated from the G and H tumors in the presence of G antisera. Of these, seven were homologous. One high-molecular-weight protein (Protein b) present on the G tumor was absent from the H tumor. The H tumor contained another high-molecular-weight protein (i) that was not found on the G tumor or on normal prostate. Normal prostate revealed a pattern similar to the G tumor except that Protein b appeared to be quantitatively reduced. Precipitation in the presence of H antisera showed similar patterns except that Protein b was not detected in the G tumor and was greatly reduced in the normal prostate. Therefore, despite variable growth characteristics, there were few changes in membrane proteins between the solid tumors and between the tumors and normal prostate. Iodination of surface proteins of cultured cells from normal prostate and the G and H sublines also showed a high degree of homology. No consistent differences between cultured cell lines were noted.

Adenocarcinoma↗
PubMed1982

Comparison of receptor properties of erythrocyte membrane glycoproteins.

Membrane glycoproteins from horse, sheep, goat and bovine erythrocytes were solubilized and purified. These glycoproteins could be placed in three groups based on their degrees of glycosylation: The major bovine erythrocyte glycoprotein (BGII) had 77% sugar, the minor bovine glycoprotein (BGI) had 27% sugar and the others had approximately 50% sugar. Four of the glycoproteins aggregated in a uniform way in aqueous solution--one, BGII, did not. Four had similar subunit sizes of 25-34,000 daltons, but BGII was larger--55,000 daltons. Receptor functions (for plant and invertebrate lectins, antibodies and cell surfaces) of these glycoproteins were primarily those dependent upon the presence of terminal sialic acid residues.

Animals↗
PubMed1980

Latex test for serodiagnosis of infectious mononucleosis.

A glycoprotein was isolated from bovine erythrocytes which has 20% carbohydrate and migrates on sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a single band. This glycoprotein carries the reactivity of bovine erythrocytes with Paul-Bunnell heterophile antibody of infectious mononucleosis. This bovine glycoprotein was coupled to carboxyl-modified latex particles with water-soluble carbodiimide. The resulting reagent was then used to develop a new test for the detection of infectious mononucleosis antibody. The bovine erythrocyte glycoprotein-latex reagent is more stable than sheep or horse erythrocytes, the traditional reagents for detection of infectious mononucleosis antibody. This new reagent is used in a direct slide test; no preabsorption of the sera is necessary. In the present study the glycoprotein-latex reagent compared favorably in terms of sensitivity and specificity with two standard tests for infectious mononucleosis antibody. Ninety-nine serum samples were tested. Agreement of the latex test with a stabilized horse erythrocyte spot test was 90%. Ten samples were weakly positive with the latex test and negative with the horse cell test. Only one of these was also positive with an enzyme-treated sheep cell test. This latter test was somewhate more sensitive than the latex test.

Animals↗