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Biomedical subjects

M Butler

Publications and source records attributed to M Butler.

At least 37 records · Page 2Linked to original sources

Comparability of old and new World Health Organization criteria for definite myocardial infarction.

New epidemiological criteria for definite myocardial infarction are now in use as part of the international study of trends and determinants of cardiovascular disease coordinated by the World Health Organization (MONICA Project). In this paper we use data from a myocardial infarction register to assess the comparability of the old and new World Health Organization criteria for definite myocardial infarction. The new criteria were applied to 905 cases who had been categorized as definite myocardial infarction on the old criteria. 739 (82%) of these cases met the new criteria for definite myocardial infarction; a further 157 cases (17%) met the new criteria for possible myocardial infarction. This study indicates that the two sets of criteria produce results that are too dissimilar for reliable comparisons to be made between studies using different criteria for definite myocardial infarction.

Adult

Alzheimer's neurofibrillary tangles contain unique epitopes and epitopes in common with the heat-stable microtubule associated proteins tau and MAP2.

Ten monoclonal antibodies raised against Alzheimer's neurofibrillary tangles (ANTs) were characterized for reactivity with heat-stable microtubule fractions from bovine and human brain. Five of the antibodies showed very little reaction, but the other five reacted strongly with heat-stable microtubule associated proteins (MAPs). The proteins recognized by these antibodies have estimated molecular weights similar to those of known heat-stable MAPs, tau (52-68 kd) and MAP2 (200-250 kd). That the proteins are indeed tau and MAP2 is demonstrated by reaction of electroblotted proteins with antibodies raised in mouse and guinea pig against bovine brain tau and MAP2. One anti-ANT antibody reacts only with tau, two bind strongly to tau and weakly to MAP2, one recognizes both tau and MAP2 equally well, and one primarily stains MAP2. Extraction of ANT with 2% SDS does not remove tau or MAP2 epitopes from ANT, indicating that epitopes shared with heat-stable MAPs are integral components of ANT. The existence of tau epitopes in ANT is also demonstrated by immunoblotting of ANT-enriched fractions with anti-tau antibodies. Most of the material recognized by anti-tau antibodies in ANT-enriched fractions is present in large molecules excluded by 3% polyacrylamide gel upon electrophoresis. Anti-tau antibodies immunostain ANT in immunofluorescence and immunoperoxidase studies. The immunostaining can be blocked by absorption of anti-tau antibodies with purified tau proteins from bovine brain. Not all ANTs in any given tissue section or isolated Alzheimer perikarial preparations, however, are stained by anti-tau antibodies. These results are consistent with previous studies that have demonstrated heterogeneity of ANTs. Whether this heterogeneity is due to biochemical modification of MAPs or absence of MAPs in some ANTs is unknown. The significance of what appear to be shared epitopes recognized by monoclonal antibodies in tau and MAP2, and the implications this may have on the pathogenesis of ANT formation, requires further investigation.

Alzheimer Disease

Microheterogeneity of microtubule-associated tau proteins is due to differences in phosphorylation.

We have studied the heterogeneity of the microtubule-associated tau proteins using tau-specific antibodies and two-dimensional electrophoresis. Both monoclonal and polyclonal antibodies to tau proteins recognize five bands in cow brain microtubule proteins run on sodium dodecyl sulfate (SDS)-polyacrylamide gels, with apparent molecular weights between 56,000 and 66,000. Immunoblots of cow brain microtubules separated on two-dimensional gels, using nonequilibrium pH gradient electrophoresis in the first dimension and SDS-gel electrophoresis in the second, reveal that greater than 30 isoforms of tau exist. The tau proteins vary in pI from 6.5 to 8.5, with the higher-molecular-weight forms being more acidic. The microheterogeneity of tau is not induced by cycling of microtubules, because two-dimensional immunoblots of tau from total brain are almost identical to those of tau from cycled tubules. Adult rat brain tau, which appears as three doublet bands on SDS gels, also exhibits considerable isoelectric heterogeneity, as does tau from 7-day-old rats, which appears as only one band on SDS gels. After dephosphorylation of cow brain tau with alkaline phosphatase, the highest-molecular-weight band disappears on SDS gels. On two-dimensional gels, the number of tau variants decreases by more than half after dephosphorylation, and the more basic species increase greatly in intensity. Preliminary experiments with tau labeled in vivo with 32PO4 also indicate that the more acidic tau proteins are the more highly phosphorylated forms. Thus, isoelectric heterogeneity of tau proteins exists at all ages and is due, at least in part to differences in the state of phosphorylation of tau isoforms.

Alkaline Phosphatase

Immunoreactive calcitonin in amyloid fibrils of medullary carcinoma of the thyroid gland. An immunogold staining technique.

A lymph node containing metastatic medullary carcinoma of the thyroid gland was examined immunohistochemically for the presence of calcitonin by light and electron microscopy. Electron microscopy showed dense, selective labeling of the tumor-associated amyloid fibrils and labeling of the scanty intracytoplasmic neurosecretory granules. By light microscopy, a few tumor cells showed strong staining, whereas the amyloid fibrils showed equivocal staining.

Amyloid

Presence of fibronectin in articular cartilage in two animal models of osteoarthritis.

Fibronectin content was determined in articular cartilage in a spontaneous dog model and in a meniscectomy rabbit model of osteoarthritis. Determination of the fibronectin content of urea extracts of articular cartilage by an enzyme linked immunosorbent assay (ELISA) disclosed that degenerated cartilage contained from 10- to 40-fold more fibronectin than normal cartilage. The finding that cartilage fibronectin content was increased in both animal models suggests that elevated cartilage fibronectin content is a general feature of the osteoarthritic process. Immunoperoxidase studies disclosed that fibronectin was distributed throughout the matrix in hyaluronidase treated normal and osteoarthritic cartilage from both animal models, but quantitative differences in fibronectin were not observed by these techniques.

Animals

A comparison of bipolar and monopolar diathermy probes in experimental animals.

The effects of monopolar and bipolar diathermy were studied in laboratory animals. The power required to coagulate transected vessels in air was established and the effect of immersion in saline and water during electrocoagulation was investigated. Tissue heat conduction from each type of probe was measured and compared. Tissue damage was assessed by light microscopy of histochemically stained sections. The bipolar system operated at a lower power output (13 W) with less heat conduction, and was unaffected by the surrounding medium.

Animals

Growth limitations in high density microcarrier cultures.

The growth of anchorage-dependent animal cells on microcarriers has enabled treatment of these cell lines as quasi-suspension cultures allowing the production of high cell densities. Analysis of the parameters affecting the final cell yield shows that if an optimal microcarrier/cell seeding ratio is provided, the surface area for cell growth is unlikely to be limiting. The culture medium could become limiting to cell growth by nutrient depletion or through the accumulation of growth inhibitors. In batch cultures of MDCK cells the analysis of amino acid utilization showed that some amino acids are nearly completely depleted from the medium during the growth period. Glutamine in particular was rapidly consumed reflecting its likely role as a major energy source. The use of a novel perfusion system for such cultures produces much higher cell densities. In 500 ml cultures perfused at 1 ml/min most of the amino acids maintained a steady concentration. The glutamine concentration was reduced not completely depleted. Under these conditions the ammonia concentration of the medium increased to a value of 2.3 mM when cell growth ceased. This level of ammonia accumulation occurred in both perfused and unperfused cultures. Investigations of the parameters affecting the growth of BHK cells in microcarrier cultures showed the rapid utilization of glutamine which was optimal at an initial concentration of 4 mM. Glucose showed rapid but not complete utilization in these experiments. The ammonia accumulation in the medium was shown to be directly related to the initial glutamine concentration and under optimal conditions rose to a level of 2.5 mM. This level of ammonia was shown to be growth inhibitory when added to cultures during inoculation. Growth experiments using culture medium diluted with isotonic salt solutions showed that the final cell yield was not directly related to the medium dilution in either perfused or unperfused cultures. This suggests that the accumulation of a growth inhibitor is responsible for growth limitation rather than nutrient depletion. The measurements of ammonia accumulation suggest that this could be the growth inhibitor which limits the final cell yield.

Amino Acids

Inactivation of human rotavirus, SA11 and other enteric viruses in effluent by disinfectants.

A preparation of infectious human rotavirus, isolated from faeces and resuspended in wastewater effluent, was shown to be inactivated by chlorine, chlorine dioxide, ozone and peracetic acid. Infectivity was assayed in MA 104 cells by the detection of cell-associated viral antigen by immunofluorescence. The inactivation curves were similar to those reported for other enteric viruses. Human rotavirus was at least as resistant as poliovirus, coxsackievirus, echovirus and f2 coliphage and was strikingly less sensitive to inactivation than the simian rotavirus, SA11. The latter was generally the most sensitive of the six tested viruses yet is often taken as being representative of the human rotaviruses.

Chlorine

The detection of rotaviruses in products of wastewater treatment.

Rotaviruses present in products of wastewater treatment were assayed in MA104 cells by indirect immunofluorescence. Levels in settled sewage, activated sludge and effluent were greater than 10(3) per litre in March and April but virus was not detected during later months. This pattern correlated with the decline in laboratory reports of human rotavirus infection.

Enzyme-Linked Immunosorbent Assay

Membrane fluidity and fatty acid composition of phospholipids in erythrocyte membranes of patients with Huntington disease.

A study was undertaken to determine whether alterations in the configuration of erythrocyte membranes of patients with Huntington disease, reported by other investigators, could be reflected in membrane fluidity-viscosity. Since membrane fluidity appears to be related to the fatty acid composition of membrane phospholipids, these were also examined. Membrane fluidity, which was determined by measuring the intrinsic fluorescence of membranous tryptophan, was found to be normal. Although the range of fatty acid distribution within the various phospholipids tended to vary somewhat, the pattern was not significantly different from that of the controls.

Erythrocyte Membrane

The influence of operating conditions of activated-sludge treatment on the behaviour of f2 coliphage.

The behaviour of f2 coliphage during activated-sludge treatment was influenced by the temperature, flow-through-time, concentration of mixed liquor suspended solids and the virus load. The most sensitive way to detect behavioural changes was to examine the regression coefficients for the rate of uptake or loss of virus by the mixed liquor solids. This type of analysis revealed, for instance, high values when the solids concentration was high and even greater values occurred when high inocula were used. At high temperature the rate of loss of virus titre after inoculation had stopped was greater than the rate of uptake of virus during inoculation although in all other conditions uptake occurred at a greater rate than the loss of virus. The coefficients were relatively low when the flow rate was increased, when the temperature was low or when the inoculum was small. The distribution of virus between the solids and liquid fractions of the mixed liquor varied somewhat for all conditions but was notably different when (a) the plant was incubated at 5 degrees C when there was much less virus in the solids fraction than usual, and (b) when the inoculum was low and a much higher proportion of virus was found in the solids. The efficiency with which virus was removed across the plant was the least-sensitive determinant of viral behaviour and the value was about the same for most treatment conditions. However, low or high inocula did result in some increased or decreased removal of virus, respectively.

Coliphages

Effects of gold, dapsone, and prednisone on serum C-reactive protein and haptoglobin and the erythrocyte sedimentation rate in rheumatoid arthritis.

Sequential measurements of serum C-reactive protein (CRP), serum haptoglobin (Hp), and erythrocyte sedimentation rate (ESR) were made in 209 patients with rheumatoid arthritis (RA); 78 of them were treated with gold, 71 with dapsone, and 60 with prednisone. The results were expressed as proportional changes in the measurements at 28-day intervals after treatment began. The period of study was 140 days. During treatment with gold and dapsone there were statistically significant gradual and progressive falls of similar magnitude in serum CRP and ESR. During treatment with prednisone serum CRP and ESR fell abruptly by 28 days and thereafter altered little. At 140 days prednisone had had the largest proportional effect on both measurements. During gold treatment the fall in serum Hp was similar to that of the ESR. In contrast, prednisone had little effect on Hp levels despite large falls in serum CRP and the ESR. Either prednisone stimulates Hp synthesis or the divergence is an expression of the difference in type of effect between gold and prednisone on RA. The effect of dapsone on serum Hp was large and progressive; it partly reflects haemolysis and, since the haemolysis was not progressive, partly improvement in the RA. The results show the relative efficacy of the drugs and suggest that dapsone may be a useful alternative treatment for RA.

Arthritis, Rheumatoid

Blockade by WB 4101 of alpha-adrenoceptors in the rat vas deferens and guinea-pig taenia caeci.

The effectiveness of WB 4101, a recently described alpha-adrenoceptor antagonist, in blocking an excitatory and two inhibitory responses to alpha-receptor activation was studied. One of the inhibitory responses was the reduction by the selective alpha-agonist amidephrine of carbachol contractures of isolated guinea-pig taenia caeci. WB 4101 antagonised this inhibition with a Schild plot slope of 0.99 and a pA2 of 8.9. The same pA2 value was obtained for blockade of the contractile effect of amidephrine and noradrenaline on the rat vas deferens. WB 4101 was, however, several hundred times less active in antagonising the inhibitory effect of clonidine on the twitch response of the vas deferens to field stimulation. Incidental observations were that the twitch was increased by low concentrations of amidephrine, and by relatively high concentrations of WB 4101. Because of its potency and postsynaptic selectivity, WB 4101 should be useful for adrenoceptor classification.

Adrenergic alpha-Antagonists