Search PubMed⌕ Search

Biomedical subjects

J G Cook

Publications and source records attributed to J G Cook.

At least 37 records · Page 2Linked to original sources

Signal propagation and regulation in the mating pheromone response pathway of the yeast Saccharomyces cerevisiae.

Extracellular signals can affect the rate of proliferation and the state of differentiation of eukaryotic cells. Signal transduction pathways have evolved to detect these signals at the plasma membrane, transmit them through the cytoplasm and into the nucleus, and thereby generate the appropriate changes in metabolism and transcription. Much attention has been focused recently on regulatory pathways of this sort that lead to activation of a family of protein kinases known as the mitogen- or messenger-activated, or extracellular signal-regulated protein kinases (MAPKs or ERKs) because this particular class of enzyme is highly conserved among eukaryotes, as is documented here and in the accompanying reviews in this issue. The mating pheromone response pathway in a unicellular microbe, the budding yeast Saccharomyces cerevisiae, is perhaps the best understood multicomponent signaling pathway known in any eukaryotic organism, especially at the genetic level. Furthermore, structural homologs and functional analogs of the components of the yeast pheromone response pathway are recapitulated in the signaling systems present in multicellular eukaryotes. This article emphasizes recent findings and common molecular themes for understanding the organization and regulation of MAPK-dependent signaling cascades that have emerged from biochemical and genetic analysis of the mating pheromone response pathway in yeast.

Cell Cycle↗

Sex- and age-specific lungworm infection in Rocky Mountain bighorn sheep during winter.

Sex- and age-specific fecal lungworm (Protostrongylus spp.) larvae concentrations in Rocky Mountain bighorn sheep (Ovis canadensis canadensis) from the Cinnabar Winter Range in southwestern Montana were determined. All 228 fecal samples collected from bighorn sheep of known sex and age class from November to April, 1984 to 1987 contained first-stage lungworm larvae. Fecal lungworm concentrations of ewes and rams declined significantly from late fall through early spring, whereas number of lungworms in lamb feces increased as winter progressed.

Age Factors↗

A study of the hypouricaemic effect of tiaprofenic acid.

Tiaprofenic acid is a non-steroidal anti-inflammatory drug which also has hypouriceamic effect. Studies involving 10 healthy volunteers were designed to investigate the mode of this effect. We postulate that the site of action of tiaprofenic acid is at the cell membrane, the mechanism being an interference with the transport of uric acid from intra- to extra-cellular fluid thus limiting its passage into the plasma. The same mechanism acting on renal tubular cells impedes reabsorption thereby increasing uric acid clearance.

Adult↗

Whole blood osmolality.

The osmolality of plasma and heparinised whole blood samples collected from hospital patients was estimated using measurement of the depression of freezing point. There was no clinically significant difference between osmolality measurement made on either whole blood, or plasma taken from the same patient. Neither cell volume nor haemolysis was found to affect the measurement. The reproducibility of whole blood measurements was similar to that for determinations carried out on plasma. Measurement of osmolality on whole blood is quicker and cheaper and needs a smaller specimen than if serum or plasma is used.

Blood↗

Diabetic ketoacidosis due to insulin resistance treated by haemodialysis.

A 14-year-old female patient with insulin resistance is reported in whom it appeared that initially insulin breakdown was unusually rapid. Subsequently there was in addition evidence of reduced insulin sensitivity. The rapidly increasing requirement of insulin (1600 units daily), given by continuous IV infusion, was dramatically reduced by haemodialysis, and was maintained subsequently. We suggest that insulin aggregates which may have blocked insulin receptor sites, causing insulin resistance, were removed by haemodialysis.

Adolescent↗

A sensitive method for the measurement of glycosylated plasma proteins using affinity chromatography.

We describe a simple, sensitive affinity technique for the routine measurement of glycosylated plasma proteins in clinical laboratories. The commercially available phenylboronic acid gel used for the chromatography has recently been marketed as a kit for this purpose (Glycogel Test Kit, Pierce Chemical Co). The manufacturers of this kit recommend loading 200 microliters neat plasma to each 1 ml gel column. This high loading is to enable the direct measurement of protein in the bound and unbound fractions at 280 nm. This loading is consistent with 10-15 mg protein being added per ml gel. Our results show that protein levels greater than 2 mg per ml gel overload the column. Therefore we used a modification of the more sensitive Bradford procedure to measure protein. The method discriminates between normals (6.29 +/- 1.87%) and diabetic patients (12.62 +/- 3.36%) and has good precision (CV 4-6%). The results obtained correlate with the colorimetric method using thiobarbituric acid (r = 0.70) and with glycosylated haemoglobin (r = 0.82).

Blood Proteins↗

Measurement of glycosylated haemoglobins and glycosylated plasma proteins in maternal and cord blood using an affinity chromatography method.

We have used a simple affinity chromatography method to measure total glycosylated haemoglobins and glycosylated plasma proteins in maternal and cord blood at 50 normal deliveries. The affinity method gives equal weighting to glycosylated haemoglobins including haemoglobin F in cord blood. The mean values for glycosylated haemoglobins in maternal blood (6.49 +/- 1.2%) were significantly higher than those in cord blood (3.85 +/- 1.0%; p less than 0.001). The difference with glycosylated plasma proteins was less marked (maternal blood 5.61 +/- 0.9% and cord blood 4.75 +/- 0.6%; p less than 0.001). A contributory factor to these differences was the decrease in glucose concentration from 4.53 +/- 0.99 mmol/l in maternal blood to 3.59 +/- 0.8 mmol/l in cord blood. The results obtained at the birth of six children to diabetic mothers showed the same trends although the mean values for glycosylated haemoglobins (maternal blood 9.27 +/- 2.3%, cord blood 4.21 +/- 0.9%), glycosylated plasma proteins (maternal blood 7.44 +/- 1.6%, cord blood 5.45 +/- 1.7%) and glucose (maternal blood 10.22 +/- 7.3 mmol/l, cord blood 5.18 +/- 3.4 mmol/l) were higher in all samples than for the deliveries to non-diabetic mothers.

Adult↗

An inexpensive, rapid and precise affinity chromatography method for the measurement of glycosylated haemoglobins.

We have assessed an affinity chromatography technique, using commercially available materials, for the estimation of total glycosylated haemoglobin in the routine clinical chemistry laboratory. The method gives good discrimination between normals (7.31 +/- 0.92%) and diabetics (12.70 +/- 2.88%) and has excellent precision (CV 1.5-2.0%). Labile glycosylated haemoglobin is normally removed as it is so variable. There is no significant correlation between labile glycosylated haemoglobin and blood glucose. Immediate analysis of incubated haemolysates is preferable to storage of haemolysates or erythrocytes. The affinity gel can be reused about 16 times, but oxidation must be reduced by keeping the gel at 4 degrees C in the dark when not in use. The cost of the gel is about 7p a test and 60 samples can be analysed in a working day. The method is not affected by the presence of up to 20% met-haemoglobin and should also give correct values for samples containing genetic variants of haemoglobin.

Anticoagulants↗

Measurement of glycosylated haemoglobins using an affinity chromatography method.

After removal of the labile material, we have measured the stable glycosylated fraction of haemoglobin with a new, commercially available, phenylboronic acid affinity gel, Glycogel B. The mean value was established for 61 non-diabetics as 7.31 (SD +/- 0.92)% and for 108 diabetics as 12.70 (SD +/- 2.88)%. The method is highly reproducible with a coefficient of variation below 2.0%. The effect of changing the temperature from 7 degrees C to 37 degrees C, and pH from 8.1 to 8.9 was investigated. For accurate results the temperature should be maintained between 20 degrees C +/- 1 degree C, and the pH between 8.6 +/- 0.1. A poor, but significant correlation (r = 0.43) between glycosylated haemoglobin and simultaneous blood glucose was shown. There was a good correlation with the agar gel electrophoretic method (r = 0.95). The slope of the regression line was 1.20 which indicates that this affinity method measures more than just HbA1. The affinity method appears to offer selectivity for diabetics than the electrophoretic method.

Blood Glucose↗

Application of analytical methods to automatic analysers.

In adapting methods for automatic analysers modifications to the chemistry may help to simplify machine design. These modifications are acceptable only if they do not adversely influence the accuracy and reproducibility of the method. The strict timing sequence which is ensured in automatic analysers, particularly of the discrete type, allows a simplification of some methods not possible in manual assays.

Autoanalysis↗