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G B Wisdom

Publications and source records attributed to G B Wisdom.

At least 19 recordsLinked to original sources
PubMed1997

Identification of alpha 1-acid glycoprotein (orosomucoid) in human synovial fluid by capillary electrophoresis.

Capillary electrophoresis of human synovial fluid in a phosphate borate run buffer containing sodium dodecyl sulphate separates a hydrophilic glycoprotein, hyaluronan and a number of low-molecular-mass components. The hydrophilic glycoprotein is identified as alpha 1-acid glycoprotein (AGP), orosomucoid, by co-injection methods with human AGP and by reaction with neuraminidase which released N-acetylneuraminic acid. Finally, a sample of the glycoprotein was isolated by micropreparative capillary electrophoresis, examined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis methods and shown to give a positive reaction with AGP antibodies. The peak due to AGP in the capillary electrophoresis is broad and gives evidence for the presence of glycoforms.

Antibodies↗
PubMed1995

The purification and properties of phosphonoacetate hydrolase, a novel carbon-phosphorus bond-cleavage enzyme from Pseudomonas fluorescens 23F.

A novel, inducible, carbon-phosphorus bond-cleavage enzyme, phosphonoacetate hydrolase, was purified from cells of Pseudomonas fluorescens 23F grown on phosphonoacetate. The native enzyme had a molecular mass of approximately 80 kDa and, upon SDS/PAGE, yielded a homogenous protein band with an apparent molecular mass of about 38 kDa. Activity of purified phosphonoacetate hydrolase was Zn2+ dependent and showed pH and temperature optima of approximately 7.8 and 37 degrees C, respectively. The purified enzyme had an apparent Km of 1.25 mM for its sole substrate phosphonoacetate, and was inhibited by the structural analogues 3-phosphonopropionate and phosphonoformate. The NH2-terminal sequence of the first 19 amino acids displayed no significant similarity to other databank sequences.

Alkaline Phosphatase↗
PubMed1994

Inhibition of fibrin monomer polymerisation by myeloma immunoglobulin.

Myelomatosis was diagnosed in a 64 year old man on the basis of a serum paraprotein band (type IgG lambda, 42 g/l), plasma cell infiltration of bone marrow, and multiple lytic lesions evident on skull x ray picture. Blood specimens taken into plain glass tubes showed bulky gelatinous clot formation with minimal clot retraction. Coagulation tests were significantly abnormal with an increase in thrombin time, prothrombin time, and reptilase time. The possibility that the paraprotein was interfering with fibrin production was investigated. The rate of fibrin monomer polymerisation (measured turbidometrically) was reduced in patient plasma compared with control plasma. Although purified fibrin monomer prepared from the patient's fibrinogen polymerised normally, the addition of purified paraprotein caused a dose dependent reduction in the rate of polymerisation. These results suggest that the paraprotein was impairing fibrin formation by inhibiting fibrin monomer polymerisation. After chemotherapy the paraprotein concentration decreased and the coagulation results returned to normal.

Blood Coagulation Tests↗
PubMed1994

Protein blotting.

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Blotting, Western↗
PubMed1994

Hereditary caeruloplasmin deficiency, dementia and diabetes mellitus.

We report two brothers with complete caeruloplasmin deficiency. The brothers presented with dementia and diabetes mellitus. Twelve relatives have partial caeruloplasmin deficiency. There is no copper overload. Transmission is autosomal recessive. DNA analysis showed genetic linkage between the deficiency and various polymorphic markers flanking the caeruloplasmin gene on chromosome 3q25. This is consistent with a mutation of the caeruloplasmin gene. Caeruloplasmin catalyses the oxidation of ferrous iron to ferric iron. Both brothers have low serum iron and increased liver iron. The index patient was given caeruloplasmin-containing, fresh-frozen plasma. A dose of 2.6 mg caeruloplasmin increased serum iron from 5 microM/l to 10 microM/l. A dose of approximately 72 mg increased serum iron from 5 microM/l to 19 microM/l. The abnormal serum and liver iron levels, and the caeruloplasmin-induced rise in serum iron, confirm a previous suggestion that caeruloplasmin maintains the normal rate of flow of iron from store to transferrin. Dementia and diabetes mellitus have been described in only one other homozygote. The absence of copper overload, and the linkage of the deficiency with chromosome 3q25, distinguish this condition from Wilson's disease.

Brain↗
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↗