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Biomedical subjects

J Newcombe

Publications and source records attributed to J Newcombe.

At least 55 records · Page 3Linked to original sources
PubMed1992

Lipocortins (annexins) 1, 2, 4 and 5 are increased in the central nervous system in multiple sclerosis.

Western blotting and densitometry have been used to investigate the lipocortin content of post-mortem central nervous system (CNS) tissue samples from multiple sclerosis (MS) patients and normal controls. Lipocortins 1, 2, 4 and 5 were all detected in normal control grey and white matter. In white matter samples from MS patients these lipocortins were found to be significantly increased, a further elevation in lipocortin content was observed in MS plaque tissue. The implications of these findings with respect to the role of these proteins in inflammatory CNS disease and a possible mechanism of steroid action in the therapy of MS are discussed.

Adult↗
PubMed1992

Measurement of low-molecular-weight antioxidants, uric acid, tyrosine and tryptophan in plaques and white matter from patients with multiple sclerosis.

The levels of the antioxidants ascorbic acid, cysteine, reduced glutathione and alpha-tocopherol, of the free-radical marker uric acid and of the amino acids tyrosine and tryptophan were measured by means of high-pressure liquid chromatography in plaques, adjacent white matter and distant white matter from patients with multiple sclerosis, and in central nervous system tissue from patients without neurological diseases. Cholesterol and DNA were also determined, to check demyelination and cellularity. Uric acid was increased and glutathione correspondingly decreased in plaques; alpha-tocopherol was lowest in plaques and highest in distant white matter in all cases. Ascorbic acid, cysteine, tyrosine and tryptophan were not significantly changed in any tissue. The results provide evidence supporting the involvement of free radicals in multiple sclerosis.

Adult↗
PubMed1991

Histopathology of multiple sclerosis lesions detected by magnetic resonance imaging in unfixed postmortem central nervous system tissue.

Postmortem unfixed whole brains from 17 multiple sclerosis and 6 control cases were examined by magnetic resonance imaging (MRI) using a T2-weighted spin echo sequence and histology to determine the relationship between areas of abnormal MRI signal and underlying pathological change. In group 1, small MRI lesions and correspondingly small plaques, most of which were chronic, were detected histologically in 5 brains. In 4 brains there were more extensive areas of both abnormal signal and histological plaques which were more often active (group 2). However, in a further 5 brains extensive MRI abnormalities were observed when only small periventricular plaques were present histologically (group 3). Lesions in the hindbrain and cerebral grey matter were infrequently observed by MRI. The extensive MRI abnormalities seen in areas in which only small histological plaques were found may be the result of vascular permeability changes in the normal-appearing white matter surrounding plaques.

Autopsy↗
PubMed1990

Spleen versus CNS immunoglobulin G in multiple sclerosis: an isoelectric focusing study.

Immunoglobulin G (IgG) band patterns were investigated in a peripheral immune compartment, the spleen, and the central nervous system (CNS) compartment in 6 multiple sclerosis (MS) subjects and 3 controls in a search for systemic humoral immune response that may be related to the localized immune response seen in MS. Using the isoelectric focusing (IEF) technique with immunoperoxidase staining of proteins transferred to nitrocellulose membranes, we were not able to demonstrate any qualitative abnormalities of spleen or serum IgG in the MS cases, whilst all matched CNS tissues and CSF specimens were clearly positive for oligoclonal IgG bands. Our results suggest that there is no systemic oligoclonal IgG secretion in MS. This further supports the presence of a compartmentalized IgG intrathecal immune response in MS and emphasizes the organ specificity of MS.

Central Nervous System↗
PubMed1988

Monoclonal antibody 14E identifies the oligodendrocyte cell body in normal adult human and rat white matter.

A monoclonal antibody, 14E, which detects immunocytochemically the perikarya of interfascicular oligodendrocytes in sections of normal adult human and rat white matter, was obtained using isolated normal human oligodendrocytes as the immunogen. 14E is an IgM antibody that reacts with a polypeptide of approximate molecular weight 23,000 on immunoblots of the particulate fraction of normal white matter. In sections of active multiple sclerosis plaques and recent infarcts, 14E immunostained the cell bodies and processes of only a small number of hypertrophic glial cells which could be oligodendrocytes undergoing pathological changes or a subpopulation of reactive astrocytes.

Animals↗
PubMed1988

A new epitope on human myelin basic protein arising from cleavage by a metalloendoprotease associated with brain myelin membranes.

Human brain myelin membranes were incubated to allow activity of an associated metalloendoprotease which cleaves myelin basic protein (MBP). A 10.3 kDa C-terminal fragment of MBP, peptide C, isolated from the incubation medium had a blocked N-terminal. After treatment with pyroglutamyl aminopeptidase, N-terminal sequencing indicated that Gln74 of MBP formed the N-terminal residue of peptide C. A rabbit antiserum was raised to a synthetic peptide containing the sequence Pyroglu-Lys-Ser-His-Gly-Arg, corresponding to the first six residues of peptide C. By immunoblotting this serum reacted with peptide C but not with intact MBP. The data indicate that cleavage of MBP by a myelin-associated protease engenders a new epitope.

Amino Acid Sequence↗
PubMed1988

The nature of inflammatory components during demyelination in multiple sclerosis.

Immune reactivity in the central nervous system (CNS) in multiple sclerosis (MS) can be fuelled by activated T lymphocytes sequestered in the brain and those entering systemically. The perivascular cuff of the inflammatory lesion consists predominantly of T cells and macrophages, while the hypercellular interface between normal and degenerating myelin is comprised mainly of macrophages and activated microglia. MHC class II+ cells are abundant in the hypercellular zone and expression of both the polymorphic and invariant chains of the molecule are expressed beyond the plaque edge in the white matter where the staining is restricted largely to microglia. Under carefully controlled staining conditions expression of MHC class II can be demonstrated on microglia in control white matter. Thus the immunological privilege of the CNS does not appear to preclude constitutive expression of MHC antigens or prevent the traffic of activated lymphocytes into the brain parenchyma. However, the antigens priming the immune reaction in the CNS in MS and the role of antibodies in the demyelinating process remain a matter for speculation and the exact mechanisms of demyelination are as yet unresolved.

Animals↗
PubMed1986

Distribution of glial fibrillary acidic protein in gliosed human white matter.

Glial fibrillary acidic protein (GFAP) in gliosed white matter from multiple sclerosis plaques and cerebral infarcts was examined by polyacrylamide gel electrophoresis and immunoblotting. Using a monoclonal antibody raised against human GFAP, up to 11 GFAP polypeptide bands of molecular weight 37-49 kilodaltons were identified in particulate and supernatant fractions of CNS tissue homogenates. Soluble GFAP constituted about one-quarter of the total GFAP in normal cerebral white matter. In brain lesions in which reactive astrocytes were observed microscopically, the proportion of soluble GFAP was increased, with a greater representation of the lower-molecular-weight forms. In brain chronic sclerotic plaques, almost all of the GFAP was in the particulate form. Purified particulate GFAP was susceptible to proteolysis at acid but not at neutral pH in the presence of CNS homogenates. In tissue autolysis studies, GFAP was stable in situ for periods well in excess of average CNS postmortem times.

Astrocytes↗
PubMed1985

Serum antibodies against central nervous system proteins in human demyelinating disease.

An immunoblotting technique has been used to screen serum samples from patients with demyelinating disease for antibody directed against central nervous system proteins. Antibodies of the IgM, IgG and IgA class directed against one or more of the particulate fraction proteins tubulin, myelin basic protein, 69 K neurofilament protein, glial fibrillary acidic protein, myelin associated glycoprotein or Wolfgram protein were present in 94, 54 and 47%, respectively, of multiple sclerosis sera examined. IgM antibodies against tubulin and myelin basic protein predominated. A similar antibody spectrum was seen in a significant proportion of sera from patients with optic neuritis, subacute sclerosing panencephalitis and motor neurone disease, in which primary or secondary demyelination occurs. Antibodies of all three classes directed against the 169 K and 220 K neurofilament proteins and against some unidentified proteins of human peripheral nerve, kidney, liver, spleen and skeletal muscle were detected in sera from healthy subjects and patients with neurological disease.

Adolescent↗
PubMed1982

Rapid analysis of immunoglobulin isoelectric focusing patterns with cellulose nitrate sheets and immunoperoxidase staining.

Immunoglobulins which have been focused to their isoelectric points in thin-layer polyacrylamide gels can be transferred by diffusion to the surface of cellulose nitrate sheets with retention of the original focusing patterns. IgG can be detected with peroxidase-linked antiserum. Unlike polyacrylamide gel, cellulose nitrate offers no permeability barrier, and so washing times are reduced from days to minutes. Thus small quantities (less than 1 microgram) of IgG may be visualized within 24 h of isoelectric focusing. In addition after immobilization on cellulose nitrate, immunoglobulin reactivity with antigens, including those of high molecular weight, may be analysed.

Animals↗
PubMed1982

Analysis of immunoglobulin G in multiple sclerosis brain: quantitative and isoelectric focusing studies.

Immunoglobulin G (IgG) in soluble and particulate fractions of discrete tissue samples from multiple sclerosis (MS) brain was analysed. Supernatant IgG/albumin rations and particulate-bound IgG levels were highest in samples dissected from MS plaques and adjacent white matter. Acid extracts of particulate fractions from the equivalent of 1 g of MS plaque tissue contained up to 15 micrograms IgG, an order of magnitude less than the amount in extracts from a subacute sclerosing panencephatitis (SSPE) brain, but 20 times more than those from control brain. By contrast, supernatant fractions from SSPE brain and some MS plaques contained comparable amounts (100-200 micrograms/g tissue) of IgG, which were 10 times greater than those from control brain. Samples were subjected to isoelectric focusing (IEF), and IgG was visualized by immunoperoxidase staining. The IEF patterns of IgG from control samples were diffuse but samples from demyelinated MS tissue displayed distinct oligoclonal bands of IgG. A number of common IgG bands were apparent in extracts of supernatant and particulate fractions from the same plaque. The IEF spectra of plaque samples from three MS brains were different. Furthermore, quantitative variations in certain IgG bands were observed in different plaques from the same brain.

Adult↗
PubMed1981

Measurement of glial fibrillary acidic protein (GFAP) and anti-GFAP antibodies by solid-phase radioimmunoassays.

A solid-phase radioimmunoassay was developed for the detection of glial fibrillary acidic protein (GFAP) and GFAP-specific immunoglobulin G (IgG) antibodies. In antibody assays purified GFAP was absorbed onto polystyrene beads, followed by incubation in dilutions of serum. (125-I)-labelled human anti-IgG was used to quantify antibodies bound to solid-phase GFAP. GFAP in samples was measured by the inhibition of the binding of anti-GFAP antibody to solid-phase GFAP. The serum and CSF samples of 19 brain tumor, 40 multiple sclerosis and 66 control patients were assayed for anti-GFAP antibodies. The binding values in the serum and CSF samples of the brain tumor patient group were higher and the binding values in the CSF samples of the multiple sclerosis group lower than those of the control group. The differences were statistically significant.

Antibodies↗