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R Simpson

Publications and source records attributed to R Simpson.

At least 19 recordsLinked to original sources

Integrin expression by human articular chondrocytes.

Expression of integrins, a family of cell adhesion proteins, by human articular cartilage chondrocytes in vivo has been studied by immunohistological techniques using cryostat sections and a panel of anti-integrin monoclonal antibodies. Chondrocytes strongly expressed beta 1 and alpha 5 integrin subunits, known to form the classical fibronectin receptor, VLA-5, strongly. alpha 1 was expressed more weakly and variably; alpha 3 was expressed by occasional cells only. Chondrocytes did not express beta 2, beta 3 or other integrin alpha subunits tested.

Antigens, CD

Sexual problems.

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Adult

Purification and partial amino acid sequence of human aconitase.

Aconitase has been purified from membranes prepared from both the human gastric carcinoma cell line Okajima and from porcine gastric mucosa by chromatography on concanavalin A-Sepharose and carboxymethyl-Sepharose, and preparative polyacrylamide gel electrophoresis. Automated Edman degradation of the intact proteins yielded no N-terminal amino acid sequence due, presumably, to N-terminal blockage. Sequence analysis of tryptic peptides derived from S-carboxymethyl porcine and human aconitases established the positions of 95 and 64 amino acid residues, respectively. The amino acid sequence data for porcine aconitase was in perfect agreement with the previously reported cDNA-deduced amino acid sequence [Zheng et al. (1990) J Biol Chem 265:2814-2821]. Comparison of the human amino acid sequence data with the cDNA-deduced amino acid sequence of porcine aconitase indicated that these two proteins have 95% amino acid sequence identity within the sequenced region.

Aconitate Hydratase

The effects of different formulations of oral contraceptive agents on lipid and carbohydrate metabolism.

BACKGROUND: Oral contraceptives can induce changes in lipid and carbohydrate metabolism similar to those associated with an increased risk of coronary heart disease, including increased serum triglyceride, low-density lipoprotein (LDL) cholesterol, and insulin levels and decreased high-density lipoprotein (HDL) cholesterol levels. In this study, we examined whether modification of the type or dose of progestin in oral-contraceptive preparations diminishes these changes. METHODS: We measured plasma lipoprotein levels and performed oral glucose-tolerance tests in a cross section of 1060 women who took one of nine types of oral contraceptives for at least three months and 418 women who took none. Seven of the contraceptive formulations contained various doses and types of progestin: levonorgestrel in low (150 micrograms), high (250 micrograms), and triphasic (50 to 125 micrograms) doses; norethindrone in low (500 micrograms), high (1000 micrograms), and triphasic (500 to 1000 micrograms) doses; and a new progestin, desogestrel, in one dose (150 micrograms). All seven contained 30 to 40 micrograms of ethinyl estradiol. Two additional formulations contained progestin alone. RESULTS: As compared with controls, women taking combination drugs did not have increased serum total cholesterol levels but did have increases of 13 to 75 percent in fasting triglyceride levels. Levels of LDL cholesterol were reduced by 14 percent in women taking the combination containing desogestrel and by 12 percent in those taking low-dose norethindrone. Levels of HDL cholesterol were lowered by 5 percent and 16 percent by the combinations containing low-dose and high-dose levonorgestrel, respectively; these decreases were due to reductions of 29 percent and 43 percent, respectively, in the levels of HDL subclass 2. The combination pill containing high-dose norethindrone did not affect HDL cholesterol levels, whereas that containing low-dose norethindrone increased HDL cholesterol levels by 10 percent. The desogestrel combination increased HDL cholesterol levels by 12 percent. Levels of apolipoproteins A-I, A-II, and B were generally increased by combination drugs. Depending on the dose and type of progestin, combination drugs were associated with plasma glucose levels on the glucose-tolerance test that were 43 to 61 percent higher than in controls, insulin responses 12 to 40 percent higher, and C-peptide responses 18 to 45 percent higher. Progestin-only formulations had only minor metabolic effects. CONCLUSIONS: The appropriate dose and type of progestin may reduce the adverse effects of oral contraceptives on many metabolic markers of risk for coronary heart disease. Progestin-only formulations or combinations containing desogestrel or low-dose norethindrone were associated wtih the most favorable profiles.

Adolescent

Liquid chromatography/time-of-flight mass spectrometry with high-speed integrated transient recording.

High-performance liquid chromatography (HPLC) has been interfaced to a time-of-flight mass spectrometer. The interface is a continuous flow probe and ions are desorbed from the liquid matrix by energetic ion bombardment. Pulsed and delayed ion extraction from the source permits the use of broad ionization times, results in the production of analog signals in each time-of-flight cycle, and provides both energy and spatial focusing. A high-speed integrated transient recording system has been developed and is also reported. This instrument is the prototype for development of a high-speed, high-mass range LC detector with high duty cycle. Its performance is demonstrated for the separation of several mixtures of small peptides.

Amino Acid Sequence

Sequence of the 3' half of the Murray Valley encephalitis virus genome and mapping of the nonstructural proteins NS1, NS3, and NS5.

We have determined the nucleotide sequence of the 3'-terminal half of the RNA genome of Murray Valley encephalitis virus (MVE) using seven overlapping cDNA clones; an estimated 80-90 nucleotides at the extreme 3'-end remain to be sequenced. In conjunction with previous sequence data for the 5' half (16), we can conclude that the MVE genome contains a long open reading frame of 10,302 nucleotides that encodes a polyprotein of 3434 residues. Comparison of the MVE deduced amino acid sequence with that of other flaviviruses shows that MVE is most closely related to Japanese encephalitis virus, consistent with serological studies. Using N-terminal amino acid sequence analysis, three nonstructural proteins (NS1, NS3, and NS5) have been identified and mapped on the MVE genome. MVE NS3 contains sequence motifs suggesting that its amino terminus may function as a serine protease. The central region of NS3 (in the linear amino acid sequence) has motifs that are found in NTP-binding proteins and helicases. MVE NS5 contains a conserved Gly-Asp-Asp sequence that is thought to be essential for RNA-dependent RNA polymerases.

Amino Acid Sequence

Human granulocyte-macrophage colony-stimulating factor (hGM-CSF): identification of a binding site for a neutralizing antibody.

One approach to the localization of functionally active regions of human granulocyte-macrophage colony-stimulating factor (hGM-CSF) is to map the epitopes recognized by neutralizing anti-hGM-CSF monoclonal antibodies. We have defined the epitope recognized by one neutralizing antibody (LMM102) using proteolytic fragments obtained by enzymic digestion of bacterially synthesized hGM-CSF. RP-HPLC fractionation of a tryptic digest resulted in the identification of an immunoreactive "tryptic core" peptide containing 66 amino acids (52% of the protein). Further digestion of this "tryptic core" with S. aureus V8 protease produced a unique immunoreactive hGM-CSF product comprising two peptides, residues 86-93 and 112-127, linked by a disulfide bond between residues 88 and 121. The individual peptides, generated by reduction with dithiothreitol, were not recognized by the antibody. An analog of this peptide has been synthesized chemically and shown to have similar immunoreactivity to the epitope obtained by enzymic digestion. A series of modified peptides has also been synthesized to identify further the region required for antibody recognition.

Amino Acid Sequence

Stopped-flow kinetics of oxygen uptake and release by embryonic red blood cells.

The processes of O2 uptake and release by the three embryonic haemoglobins contained within early mouse embryonic red blood cells have been studied using dual-wavelength stopped-flow kinetic spectroscopy. The rate of O2 uptake in the pseudo-spherical, nucleated, embryonic red blood cells exhibits a greater than first-order dependence on O2 concentration. The time courses for the release from the red blood cells into dithionite-containing solutions tends towards a limiting rate at high dithionite concentrations. The rates of both the uptake and release processes observed in the embryonic cells are compared with those previously seen for adult mouse red blood cells. A new mathematical model is described which accurately simulates both uptake and release experimental data for the nucleated embryonic red blood cells.

Animals

An analysis of the stopped-flow kinetics of gaseous ligand uptake and release by adult mouse erythrocytes.

Ligand uptake and release by the haemoglobin contained within adult mouse erythrocytes was studied by using dual-wavelength stopped-flow techniques. The rate of O2 uptake is very much lower than that expected for an equivalent concentration of haemoglobin in free solution. The O2-concentration-dependence found in uptake experiments is greater than first-order. CO uptake shows the same pattern of reactivity as does O2, but the associated rates of uptake are lower and the concentration-dependence of the CO rates is first-order. O2 release from the adult erythrocytes was measured by stopped-flow mixing with Na2S2O4. Under these circumstances the deoxygenation of intracellular haemoglobin shows accelerating time courses. The apparent rate-constant-dependence on dithionite concentration shows a rate limit at high reductant concentrations. Computer simulations of both ligand uptake and release processes were carried out by using a three-dimensional model. The simulations clearly indicate that in rapid-mixing experiments the rather slow experimentally observed O2 uptake rate is due to rate-limiting diffusion through an extracellular stagnant solvent layer. In the case of O2 release, however, the major rate-controlling process is the rate of O2 dissociation from the haemoglobin molecules, which accelerates during the deoxygenation process.

Animals

Antibodies to neutrophil cytoplasmic antigens in Wegener's granulomatosis and other conditions.

The use of serum antibodies to neutrophil cytoplasmic antigens (ANCA) as a diagnostic marker for Wegener's granulomatosis and other forms of vasculitis has been assessed. Although ANCA have been described by several groups the precise antigenic targets are unknown, and detection of ANCA still relies on an indirect immunofluorescence assay technique. Several different patterns of fluorescence have been produced by using sera from different groups of patients, and insufficient information is available on the frequency of positive results and of the patterns of immunofluorescence obtained when serum from patients with vasculitis as a part of a generalised connective tissue disease is used. A study was carried out on serum from 240 patients, including 23 patients with Wegener's granulomatosis, 12 with microscopic polyarteritis, and 30 with various connective tissue diseases. Three patterns of fluorescence were observed: bright coarsely granular cytoplasmic, bright non-granular cytoplasmic, and weak diffuse cytoplasmic. The bright, coarsely granular pattern was 86% specific for Wegener's granulomatosis in this series and was observed in 18 of 23 cases. Other patterns of fluorescence were found in various conditions and were not of diagnostic value. The technique is simple, inexpensive, rapid, and reproducible.

Adult

Use of a controlled duodenal fistula in the management of a major duodenal injury.

The management of a major duodenal injury associated with gross peritoneal contamination caused by multiple colonic perforations is reported. Resection of devitalized duodenum with creation of a controlled duodenal fistula, a jejunostomy and three colostomies, together with subsequent reconstruction, led to a successful outcome.

Catheters, Indwelling