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Biomedical subjects

M J Walton

Publications and source records attributed to M J Walton.

12 recordsLinked to original sources
PubMed1987

Studies on the putative role of gamma-glutamyl transpeptidase in intestinal transport of amino acids in Atlantic salmon.

The gamma-glutamyl cycle is considered to function in the membrane transport of amino acids, particularly glutamine and cysteine. When groups of Atlantic salmon were fed either a control diet containing 45% crude protein or an amino acid diet (of similar overall amino acid composition but containing elevated levels of glutamine and cysteine) for 16 weeks, weight gains were significantly greater in the former group than in those given the amino acid diet. There were no significant differences between treatments in gamma-glutamyl transpeptidase (GT) activity in the proximal intestine; in distal intestine there was significantly more activity in control fish. Mean levels of GSH were higher in tissues (pyloric caeca, distal intestine and kidney) of amino acid diet fish than in those of control fish. Glutamine was less effective as a gamma-glutamyl acceptor than several other amino acids when tested with salmon caecal GT. There were no morphological adaptations to the two feeds. Nutrient uptake studies showed an increased uptake of glutamine, but decreased uptakes of proline and methionine in proximal intestine of salmon fed amino acid diet. Much the greater part of the glutamine uptake, even at high concentrations was shown to be by Na+ dependent processes. There is no evidence that GT itself is Na+ dependent. The results do not support the view that the gamma-glutamyl cycle and GT in particular are involved in the transport of amino acids in the intestine and are discussed in this context.

Amino Acids↗
PubMed1985

Intracellular distribution of tricarboxylic acid cycle enzymes in liver of rainbow trout Salmo gairdneri.

The intracellular distribution of enzymes of the TCA cycle was investigated in liver of rainbow trout. All enzymes of the cycle apart from succinyl thiokinase were detected. Citrate synthase, alpha-ketoglutarate dehydrogenase and succinate dehydrogenase were wholly mitochondrial. Fumarase, malate dehydrogenase, aconitase and NADP-isocitrate dehydrogenase were detected in both cytosol and mitochondria.

Animals↗
PubMed1984

The effects of dietary tryptophan levels on growth and metabolism of rainbow trout (Salmo gairdneri).

Groups of rainbow trout (Salmo gairdneri) (mean weight 14 g) were given diets containing 0.8, 1.3, 2, 3, 4 or 6 g tryptophan/kg diet for 12 weeks. By analysis of the growth results, the dietary requirement of tryptophan was found to be 2.5 g/kg diet (equivalent to 50 mg/kg biomass per d). Carbon dioxide expired by trout following intraperitoneal injection of [14COOH]tryptophan contained little radioactivity when dietary tryptophan level was low but, above 2.0 g/kg diet, it increased rapidly with increasing dietary tryptophan level. The break point in the dose-response curve did not, however, coincide with that from the growth results. Changes in concentrations of free tryptophan in blood and liver and activity of hepatic tryptophan pyrrolase (EC 1. 13. 11. 11) in response to changes in dietary tryptophan concentration did not provide reliable indicators for quantifying dietary requirement. Unlike the situation in mammals, blood tryptophan was not protein-bound to any appreciable extent. Tryptophan pyrrolase of trout has properties which suggest it has no apoenzyme form. In fish given adequate levels of tryptophan injected intraperitoneally with a tracer dose of [14COOH]tryptophan, 60% of the dose was incorporated into body protein within 1 d. The turnover of the label in this protein is very slow. Those trout given diets deficient in tryptophan suffered from severe scoliosis and lordosis as well as having increased liver and kidney levels of calcium, magnesium, sodium and potassium.

Animals↗
PubMed1984

The effect of dietary lysine levels on growth and metabolism of rainbow trout (Salmo gairdneri).

Groups of rainbow trout (Salmo gairdneri; mean weight 5 g) were given diets containing 10, 12, 14, 17, 21, 24 and 26 g lysine/kg diet for 12 weeks. By analysis of the growth values the dietary requirement of lysine in this experiment was found to be 19 g/kg diet. A similar requirement value was obtained from a dose-response curve of expired 14CO2 (following an intraperitoneal injection of L-[U-14C]lysine) v. dietary lysine concentration. Liver concentrations of total lipid and carnitine and activities of lysine-alpha-ketoglutarate reductase (saccharopine dehydrogenase (NADP+, lysine-forming), EC 1.5.1.8) in the liver were not significantly different in fish from the different dietary treatments. Hepatosomatic index, however, was higher in those fish given low levels of dietary lysine.

Animals↗
PubMed1982

Methionine metabolism in rainbow trout fed diets of differing methionine and cystine content.

The metabolism of methionine was studied in rainbow trout fed diets containing different levels of methionine and cystine. Growth data indicated that methionine requirement was between 0.5 and 1% dry diet in the absence of dietary cystine, but 0.5% was adequate when dietary cystine was 2%. In fish fed diets deficient in sulfur-containing amino acids, elevated hepatic activities of glutathione reductase were found, whereas glutathione peroxidase and glutathione levels were unaffected. Plasma and liver concentrations (18 hours after feeding) of methionine and cystine were affected by dietary methionine, but dietary cystine had little effect. Cystine appeared to be converted to taurine in the liver. In fish injected intraperitoneally with [14COOH]- and [14CH3]methionine, over 24 hours the carboxyl group was oxidized more than the methyl group and more was incorporated into protein. However, much more of the methyl group was incorporated into the lipid fraction. The results suggest the operation of the transsulfuration pathway of methionine catabolism. Oxidation of methionine was related to its concentration in the tissues, and little affected by dietary cystine. A 28-day experiment on the metabolism of injected [14COOH]methionine showed that its turnover was slow, and much of the radioactivity was associated with protein.

Amino Acids↗
PubMed1981

Tissue distribution, uptake, and requirement for alpha-tocopherol of rainbow trout (Salmo gairdneri) fed diets with a minimal content of unsaturated fatty acids.

The metabolism of and requirements for alpha-tocopherol in rainbow trout fed diets containing 1% linolenic acid as sole source of unsaturated fat and graded levels of tocopherol (0.06-10 mg/100 g) were examined. Fish grew 5-fold over a 16-week period. In liver, tocopherol was concentrated in mitochondria with little in cytosol. Orally administered [3H]-tocopherol was rapidly taken up by plasma and liver but uptake into erythrocytes and white muscle was much slower; in most tissues radioactivity reached a plateau after about 3 days but in red muscle radioactivity increased over a 10-day period. Activities of enzymes that prevent free radical initiated tissue damage did not change in tocopherol deficiency. Tocopherol-deficient trout had no gross or subcellular pathologies even though liver and muscle were severely depleted of the vitamin. Ascorbic acid-stimulated lipid peroxidation in liver organelles indicated a tocopherol requirement of 2-3 mg/100 g diet; the molar ratios of polyunsaturated fatty acids to tocopherol in livers of trout fed diets lacking or supplemented with tocopherol (100 mg/100 g) were 980 and 170, respectively.

Animals↗
PubMed1972

Some studies on the biosynthesis of ubiguinone, isoprenoid alcohols, squalene and sterols by marine invertebrates.

The ability of fourteen marine invertebrates to utilize [(14)C]mevalonate for the biosynthesis of isoprenoid compounds was investigated. Several of the animals, in particular crustaceans, bivalve molluscs, a coelenterate and a sponge, were unable to synthesize squalene and sterols, whereas gastropod molluscs, echinoderms, an annelid and a sponge could. Regardless of sterol-synthesizing ability the animals (with the exception of a sponge) always made dolichol and ubiquinone, and thus a specific block in squalene and sterol synthesis was indicated in some animals. Radioactivity accumulated in relatively large amounts in farnesol and geranylgeraniol in those animals incapable of making sterols.

Alcohols↗
PubMed1971

Improved chemically defined basal medium (CMRL-1969) for primary monkey kidney and human diploid cells.

An improved tissue culture basal medium, CMRL-1969, supplemented with serum, has been evaluated by measuring the growth responses of primary cultures of trypsin-dispersed monkey kidney cells (PMKC) and of an established culture of a human diploid cell strain (HDCS). Medium H597, an early modification of medium 199 which has been used successfully in the preparation of poliomyelitis vaccine for 15 years, was used for comparison. In addition, parallel testing was done with Basal Medium Eagle (BME) widely used for the growth of HDCS. The improvements in basal medium CMRL-1969 are attributed to changes in amino acid concentrations, in vitamin composition, and, in particular, to enhanced buffering capacity. The latter has been achieved by the use of free-base amino acids and by increasing the dibasic sodium phosphate. The new medium has already been used to advantage for the production of polioviruses in PMKC where equivalent titers were obtained from cultures initiated with 70% of the number of cells required with earlier media. The population-doubling time was reduced in this system. Also, with small inocula of HDCS, the time required to obtain maximum cell yield was shorter with CMRL-1969 than with BME. Both media were supplemented with 10% calf serum. Maximum cell yields after repeated subcultivation in the new basal medium were greatly increased and the stability of the strain, as shown by chromosomal analysis, was not affected. Basal medium CMRL-1969 can be prepared easily in liquid or powdered form.

Amino Acids↗
PubMedDate not supplied

Karyology: vaccine manufacturers' views.

Chromosomal analyses and life span studies of WI-38 cell cultures used for vaccine production at the Connaught Laboratories Ltd will be described. Twenty cultures derived from the same seed lot and 25 cultures derived from different starter cultures were analyzed. Karyology tests at population doublings 31 and higher were within acceptable limits. The WI-38 cells reached a finite lifespan of approximately 65 population doublings. Normal growth characteristics were observed within 2/3 of the finite lifespan, i.e. until population doubling 40. The finite lifespan of MRC-5 was found to be 75 population doublings, with normal growth pattern until passage 50. The karyology test results, cell yields and the finite lifespans indicated that these cells could be used as substrates for vaccine production at higher passage levels than previously suggested. Using serially subcultivated cells at higher passage levels would make possible an increase in the volume of vaccine from existing cell seed and would also result in significant reduction in production costs.

Cell Survival↗