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Biomedical subjects

M G Rumsby

Publications and source records attributed to M G Rumsby.

At least 55 records · Page 3Linked to original sources
PubMed1983

Chronic relapsing experimental allergic encephalomyelitis. The presence in the cerebrospinal fluid of factors chemotactic for monocytes.

Cerebrospinal fluid (CSF) was removed from guinea pigs with chronic relapsing experimental allergic encephalomyelitis (CR-EAE) and control inoculated animals by puncture of the cisterna magna. The fluid from 7/8 animals in relapse and 2/4 animals in remission phases of CR-EAE was found to promote the migration of peripheral blood monocytes through a 5-micron pore diameter polycarbonate membrane filter. Monocytes were also found to orient towards the migration-stimulating CSF in a gradient formed between such fluid and CSF derived from a control animal, thereby indicating the presence of a chemotactic factor. The factor responsible for promoting monocyte migration had a molecular weight of between 50 000 and 300 000 as defined by ultrafiltration. The results are discussed in relation to the known pathohistological features of the chronic relapsing disease.

Animals↗
PubMed1983

Chronic relapsing experimental allergic encephalomyelitis. Immunological and blood--cerebrospinal fluid barrier-dependent changes in the cerebrospinal fluid.

Cerebrospinal fluid (CSF) and plasma were taken from strain 13 guinea pigs in various stages of chronic relapsing experimental allergic encephalomyelitis using techniques which allowed repeated sampling from the same animal. Samples were assayed for albumin and IgG and the corresponding CSF/plasma quotients evaluated graphically using a method which could discriminate between blood-CSF barrier dysfunction and local IgG synthesis in the central nervous system (CNS). During the disease a 2-3-fold increase in plasma IgG concentration developed and an increase in blood-CSF permeability was noted. Isoelectric focusing revealed an oligoclonal IgG pattern identical in both plasma and CSF. The results provided no evidence for a local production of IgG in the CNS like that which is known to occur in multiple sclerosis.

Albumins↗
PubMed1982

A syringe modification for the simple and rapid dissociation of post-natal rat cerebral tissue for preparing primary cultures of mixed glial cells.

Mechanical disruption of 2-day post-natal rat cerebral tissue without enzyme pretreatment through 21-gauge and 23-gauge hypodermic needles on a 1 ml disposable plastic syringe, gives dissociated preparations which contain between 1.7 and 2.1 X 10(7) cells/100 mg wet tissue. At a plating density of between 7.1 and 9.2 X 10(5)/cm2 in culture, between 2 X 10(5) and 2 X 10(6) cells are recovered after 7 days. Use of needles finer than 23-gauge resulted in lower cell yields and loss of galactocerebroside-positive oligodendrocytes. The method described is convenient for the routine preparation of mixed primary glial cultures on a regular basis where many cerebra need to be processed.

Animals↗
PubMed1982

Effect of Co2+-substitution on the substrate specificity of phospholipase C from Bacillus cereus during attack on two membrane systems.

Phospholipid degradation by native phospholipase C from Bacillus cereus and enzyme forms where one or both of the Zn2+ prosthetic groups had been replaced with Co2+ was studied in human erythrocyte membranes (ghosts) and resuspended freeze-dried bovine brain myelin. The rate of total phospholipid degradation was 2-9-fold more rapid with erythrocytes than with myelin. With both membrane systems the activity decreased in the order ZnZn-enzyme greater than ZnCo-enzyme greater than CoCo-enzyme. For all three enzyme forms with either membrane system, phosphatidylethanolamine (or the ethanolamine-containing phosphoglycerides) and phosphatidylcholine were hydrolysed most rapidly and sphingomyelin least. The relative rate of sphingomyelin degradation was highest with the ZnCo-enzyme. In myelin at low ionic strength there seemed to be a core of phospholipid that was very resistant to degradation by native phospholipase C but which was much more accessible to the Co2+-substituted forms. It is suggested that ZnCo-phospholipase C has potential applications in membrane studies.

Animals↗
PubMed1980

Lysis of erythrocytes from stored human blood by phospholipase C (Bacillus cereus).

The ability of phospholipase C (Bacillus cereus) to lyse erythrocytes from human blood that had been stored under Transfusion Service conditions for up to 16 weeks has been examined. When incubated at 20 degrees C with enzyme (0.03 mg/ml, 55 units/ml) for up to 1 h fresh erythrocytes were not lysed. After about 4 weeks of storage a population of very readily lysed erythrocytes appeared. The morphological changes in erythrocytes from blood stored up to 16 weeks were examined by scanning electron microscopy. The proportion of very readily lysed erythrocytes correlated well with the proportion of spheroechinocytes I. This morphological form was shown to be preferentially removed by phospholipase C and before lysis a transient appearance of smooth spheres occurred. The decrease in blood ATP concentrations on storage was measured and found to correlate with the disappearance of discoid erythrocyte forms, but not directly with the increased susceptibility of the erythrocytes to lysis by the enzyme. However, erythrocytes of up to at least 15 weeks of age could be made less susceptible to lysis by pre-incubation in a medium designed to cause intracellular regeneration of ATP. During the lysis of spheroechinocytes I by electrophoretically pure recrystallized phospholipase C a rapid degradation of phosphatidylcholine, phosphatidylethanolamine and phosphatidylserine + phosphatidylinositol) occurred together with a slower degradation of sphingomyelin.

Adenosine Triphosphate↗
PubMed1980

Accessibility of galactosyl ceramides to probe reagents in central nervous system myelin.

The suitability of isolated central nerve myelin preparations for probe labelling studies was assessed and the accessibility of galactosyl ceramides in myelin to galactose oxidase and sodium periodate was determined. Isolated myelin preparations present a uniform external membrane surface to added probes because lamellae in the myelin sheath separate at their external apposition surfaces exclusively during isolation. The cytoplasmic apposition remains intact in isolated myelin. Cationised ferritin can gain access along external apposition regions of inner lamellae in multilamellar fragments of isolated myelin, indicating that proteins and lipids on the external membrane surface will be accessible to probes. Over 50% of the total galactosyl ceramides of myelin are accessible to galactose oxidase attack; hydroxy fatty acid- and nonhydroxy fatty acid-containing cerebrosides are equally attacked. Sodium periodate attacks over 90% of the galactosyl ceramides in isolated myelin at 20 degrees C and electron micrographs of the periodate-treated myelin reveal changes at the external apposition only. Galactosyl ceramides in vesicles of myelin lipid vesicles are not so readily attacked by periodate. The disposition of galactosyl ceramides in the myelin lamellae is discussed.

Animals↗
PubMed1980

Ageing of blood in hereditary spherocytosis.

Blood from 5 patients suffering from hereditary spherocytosis (HS) was stored in acid-citrate-dextrose anticoagulant at 4 degrees C for up to 6 weeks. The erythrocyte morphology and susceptibility to lysis by phospholipase C (Bacillus cereus) were examined at 2-weekly intervals and compared with erythrocytes from 6 different donations of stored normal blood. the hereditary spherocytes went through essentially the same series of morphological changes as did normal erythrocytes. Fresh hereditary spherocytes were very resistant to lysis by phospholipase C, but, like normal erythrocytes, became progressively more susceptible to lysis upon storage. In terms of erythrocyte morphological changes and development of susceptibility to lysis by the enzyme, blood from HS patients aged less rapidly than did normal blood. Splenectomy appeared to have no marked effect on the storage properties of blood from these patients as indicated by erythrocyte morphology and susceptibility to lysis by phospholipase C.

Erythrocyte Aging↗
PubMed1979

The intrinsic fluorescence of isolated central-nervous-system myelin-sheath preparations.

The intrinsic fluorescence characteristics of tyrosine and tryptophan residues in the proteins of isolated central-nervous-system myelin were investigated to gain information concerning the location of these residues within the intact membrane system. Tryptophan fluorescence from isolated myelin has an emission maximum at 325 nm that appears to arise from at least two different populations of tryptophan residues. Further evidence for heterogeneity of tryptophan location in the membrane is obtained from quenching studies with chloroform and acrylamide. It is speculated that one tryptophan population is hydrophobically situated and may be derived from the proteolipid protein of myelin, whereas the other tryptophan population is located at the membrane surface and may arise from the extrinsic basic protein. A significant tyrosine fluorescence is detected from isolated myelin, indicating that some of these residues are not quenched by structural interactions within the lipid--protein membrane system. Studies with freeze-dried resuspended myelin suggest that the structural arrangement of protein components in the dried rehydrated membrane system differs significantly from that of the freshly isolated myelin membrane.

Acrylamides↗