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Biomedical subjects

G B Wisdom

Publications and source records attributed to G B Wisdom.

At least 37 records · Page 2Linked to original sources
PubMed1992

The determination of asialoglycoforms of serum glycoproteins by lectin blotting with Ricinus communis agglutinin.

Serum proteins were fractionated by polyacrylamide gel electrophoresis under denaturing conditions and transferred to nitrocellulose membranes. The blotted polypeptides were probed with biotinylated Ricinus communis lectin (RCA120) followed by streptavidin/alkaline phosphatase. This procedure detected five asialoglycoproteins (alpha 2-macroglobulin, transferrin, alpha 1-antitrypsin, alpha 1-antichymotrypsin and haptoglobin beta chain). The asialoform of the alpha 1-trypsin inhibitor was found to be decreased in inflammation.

Alkaline Phosphatase↗
PubMed1992

Pre-beta high-density lipoprotein determined by immunoblotting with chemiluminescent detection.

We describe a novel assay of pre-beta high-density lipoprotein (HDL), a unique apolipoprotein A-I (apo A-I)-containing lipoprotein particle. The pre-beta and alpha lipoproteins are separated by electrophoresis in agarose and transferred onto a membrane by capillary blotting. The membrane blot is sequentially incubated with sheep anti-human apo A-I antiserum and then with a conjugate of rabbit anti-sheep immunoglobulin and horseradish peroxidase. Chemiluminescence formed by the peroxidase-catalyzed oxidation of luminol in the presence of an enhancer is captured on photographic film, and the pre-beta HDL band is quantified by transmission densitometry. The assay is calibrated with standards prepared from a reference serum diluted in 9 mol/L urea. Within-batch precision (CV) at pre-beta HDL concentrations of 22.1 and 44.3 mg/L was 7% and 4.9% respectively. Pre-beta HDL contained 1.6% (0.65-2.6%, mean and range) of total serum apo A-I in 30 normolipidemic subjects.

Adult↗
PubMed1991

A magnetizable solid phase for enzyme extraction.

A method for the convenient and reliable preparation of magnetizable agarose beads containing iron particles is described. The beads were treated with the triazine dye, Reactive Red 120, and the matrix was examined for the ability to extract proteins from crude preparations using lactate dehydrogenase from porcine muscle as a model. The recovery and specific activity values of enzyme obtained using this matrix and magnetic field separation were significantly greater than those for enzyme purified by centrifugation and conventional dye ligand chromatography.

Adsorption↗
PubMed1989

Antibodies to simian virus 5 in patients with multiple sclerosis and other neurological disorders.

Antibodies to the paramyxovirus simian virus 5 were measured in cerebrospinal fluid samples using an enzyme-linked immunosorbent assay. Six of the 13 clinically definite MS patients had elevated levels of antibodies compared with other neurological disease and orthopaedic controls. None of the samples from MS patients classed as probable or possible had increased amounts of SV5 antibodies. Simian virus 5 antibodies and measles antibodies showed a weak correlation and it is suggested that the elevated levels of the former are a manifestation of the increased antiviral response found in some MS patients.

Adult↗
PubMed1989

Apolipoprotein E phenotype determined by agarose gel electrofocusing and immunoblotting.

Polymorphism at the apolipoprotein E (ApoE) locus is an important factor in the development of remnant (Type III) hyperlipidemia and also influences the distribution of cholesterol concentrations in the population. The new method for ApoE phenotyping described here gives good results with simple apparatus. Serum (10 microL) is digested with sialidase (EC 3.2.1.18), delipidated, and redissolved in 6 mol/L urea. Electrofocusing is carried out in agarose, followed by immunoblotting with a monoclonal antibody to ApoE and an anti-immunoglobulin-peroxidase conjugate. Sialidase-catalyzed digestion effectively removes sialated forms of ApoE, which eases interpretation. This method can be used in nonspecialist laboratories and is particularly suited for assay of large numbers of samples.

Antibodies, Monoclonal↗
PubMed1988

Screening of multiple sclerosis cerebrospinal fluid for autoantibodies.

An enzyme-linked immunoadsorbent assay, using nitrocellulose discs as solid phase and small sample volumes (50 microliter), was developed for the measurement of antibodies. This was used to screen CSF samples for autoantibodies against tissue components. Extracts from a selection of tissues from both "normal" and MS patients and from 3 glial cell lines were made in phosphate-buffered saline; in the case of neural and lymphoid samples the remaining particulate materials were subsequently solubilised with octylglucoside. The saline-soluble components were screened against CSF samples from MS patients (18), patients with other neurological disorders (10), and matched orthopaedic patients but no differences were found among the 3 groups. However, when the detergent-soluble components were screened a significant (at the P less than or equal to 0.01 level) elevation of reactivity towards brain was found in 6/16 MS patients and 2/12 patients with other neurological diseases when compared to their controls.

Adult↗
PubMed1985

Detection of autoantigens by immunoblotting using a peroxidase-anti-peroxidase complex.

An immunoblotting procedure using a peroxidase-anti-peroxidase (PAP) complex was developed for the detection of autoantigens in crude mixtures by human autoimmune sera. Thymus proteins were transferred to a nitrocellulose sheet after electrophoresis in polyacrylamide gels and probed with a 1:100 dilution of serum. The location and extent of immunoglobulin G (IgG) binding was determined by sequential reaction with: rabbit anti-human IgG, goat anti-rabbit IgG and rabbit peroxidase-anti-peroxidase complex. The peroxidase was allowed to react with chloronaphthol and low levels of autoantigen/autoantibody complex were detectable with virtual absence of background colour. The inclusion of human IgG and its pepsin-generated fragment provided a means of controlling and calibrating the blotting procedure.

Animals↗
PubMed1985

A survey of serum antibodies to eight common viruses in psychiatric patients.

Serum antibody titres to eight neurotropic viruses were measured by enzyme immunoassay in 450 psychiatric in-patients and 143 controls. A seasonal variation in schizophrenic births was observed, with a peak incidence between March and April. Both herpes simplex virus and cytomegalovirus antibody titres correlated with age and, when this was controlled for, no significant differences emerged between any patient group and the controls. Mumps antibody titres were significantly lower in patients with mental subnormal and neurosis or personality disorder; measles and rubella antibody titres were lower in male but not female mentally handicapped patients; males had lower antibody titres to mumps, cytomegalovirus and Epstein-Barr virus than females in all groups. A decrease in mumps antibody titres was also found in schizophrenics if the medication factor was excluded. These low antibody titres may indicate an impaired immune response. Thus perinatal or childhood subclinical viral infections of the central nervous system, particularly of mumps, might lead to a range of possible psychiatric outcomes in later life.

Adolescent↗
PubMed1985

Serum and CSF antibody titres to seven common viruses in schizophrenic patients.

CSF and matched serum antibody titres to seven common viruses were measured in 20 chronic schizophrenic patients, and 17 of these were age and sex-matched with orthopaedic controls. CT scans were carried out in patients and age and sex-matched radiological controls. There was a trend for CSF viral antibody titres (except CMV, HSV and VZV) to be decreased in the patients compared to controls, statistically significant for mumps and IgG. The CSF/serum ratios showed a reduction in the patients, compared to controls, statistically significant for measles and rubella as well as mumps and IgG. Cerebral ventricular size was significantly increased in the patients compared to controls, but did not correlate with any of the antibody data. These findings suggest that there is a reduced immune response to certain common viruses in the CNS of schizophrenic patients, but possible effects of institutionalisation or current medication could only be adequately excluded by further prospective studies.

Adult↗
PubMed1984

Human umbilical cord conditioned medium: a stimulus for human CFU-G.

Segments of human umbilical cord have been used to prepare a highly active conditioned medium (HUCCM) for in vitro human granulopoiesis and have been compared with human placenta conditioned medium (HPCM) and feeder layers. The HUCCM produced more colonies on day 7 than HPCM (P less than 0.001), and colonies consisted almost exclusively (over 98%) of granulocytic cells by both morphological and cytochemical analysis. The HUCCM-stimulated colonies have a short culture life, the number of colonies being maximal on day 7 of culture with few colonies remaining on day 10. The main source of the colony-stimulating activity in the umbilical cord was found in the Wharton's Jelly component.

Colony-Forming Units Assay↗
PubMed1980

Variability in the adsorption properties of microtitre plates used as solid supports in enzyme immunoassay.

We evaluated the variability in the amount of protein adsorbed onto the surface of individual wells of an assortment of microtitre plates by use of procedures involving enzyme-protein conjugates. Coefficients of variation in adsorbed protein ranged from 5.2 to 29.5%. Microtitre plates show a distinct "edge effect"; wells at the edges of a plate adsorb more protein than those in the interior, which markedly affects results from immunoassays involving such plates. In a sandwich enzyme immunoassay the results for an individual sample varied by +/- 18%; in a competitive assay, values read from different calibration curves in the same plate varied by +/- 11%. Scanning electron microscopy and birefringence studies demonstrated no marked physical differences between individual wells on a plate. The variability in the amount of protein adsorbed onto the surface of individual wells on the same microtitre plate seriously affects the reliability and interpretation of results of immunoassays in which such plates are used as solid supports.

Alkaline Phosphatase↗