Under the banyan tree.
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Community Health Services↗
Biomedical subjects
T Richardson
Publications and source records attributed to T Richardson.
Purification and characterization of pregastric esterase from calf.
Calf pregastric esterase (PGE) was purified from calf gullet tissues. The tissue was excised and lyophilized, and lipid materials were extracted with acetone and n-butanol at -20 degrees C. Proteins were extracted from the delipidated tissue with a buffer containing a chaotropic salt (NaSCN) to solubilize hydrophobically bound protein aggregates. Calf PGE precipitated from the crude extract at pH 5.0. The precipitated, solubilized proteins were subjected to anion-exchange chromatography on DEAE-Sephacel, and the enzymatic activity was eluted using a linear gradient from 0.10 to 0.50 M NaCl at pH 8.0. Fractions with high specific activities were then chromatographed twice using gel filtration on Sephadex G-100. The resultant enzyme was shown to be pure upon discontinuous electrophoresis in 12% polyacrylamide containing 0.1% sodium dodecyl sulfate (SDS-PAGE). From SDS-PAGE gel patterns, a molecular weight of 49,000 was determined. The amino acid composition of the enzyme allowed calculation of an "average hydrophobicity" (Bigelow index) of 1150 cal/residue. This indicates that calf PGE is relatively hydrophobic, being similar to proteins such as alpha-lactalbumin and bovine serum albumin in average hydrophobicity.
Amino Acids↗
The effect of computer generated comments on the distribution of anticonvulsant concentrations.
Following the introduction of a consistent method of adding informative comments to the laboratory report of the plasma concentrations of the anticonvulsants phenytoin, carbamazepine and phenobarbitone, there was a significant increase in the proportion of results falling within the therapeutic range to 64, 65 and 62% respectively. This increase was mainly at the expense of results which previously fell below this range.
Anticonvulsants↗
Estimating fetal weights less than 2000 g: an accurate and simple method.
An accurate estimated fetal weight in the low-birth-weight infant can provide valuable information for the obstetrician faced with a decision concerning mode and time of delivery. Four existing equations for calculating fetal weight were analyzed and compared with data consisting entirely of infants with birth weights of 500-2000 g. A new formula, birth weight-grams = 10.1 (abdominal circumference-cm X biparietal diameter-cm) - 481, is proposed as a simple yet accurate alternative. This equation has a standard deviation of 99 g/kg.
Birth Weight↗
Composition and ultrastructure of calcium phosphate-citrate complexes in bovine milk systems.
The present studies show that the colloidal calcium phosphate of cows' milk has a (Ca + Mg)/Pi ratio of 1.67 (+/- 0.10; n = 22) and contains citrate, Mg and Zn at molar ratios to Ca averaging 0.05, 0.03 and 0.003, respectively. The composition of the natural colloidal phosphate of milk is similar to the precipitates formed by neutralization of ultrafiltrates obtained from acidified milks, and to that of the calcium phosphate-enriched fraction produced by extensive enzymic hydrolysis of the casein micelles in milk. Examination by electron microscopy of these artificial preparations of milk calcium phosphate revealed in both a very fine and uniform substructure which consisted of granules having an average, true diameter of approx. 2.5 nm. The size and shape of these tiny granules closely resemble the morphologies reported for the colloidal phosphate particles in native casein micelles, as well as for the subunits of amorphous calcium phosphate observed during calcification in other biological systems such as mitochondria and bone.
Animals↗
Is the lecithin/sphingomyelin ratio outdated?
The lecithin/sphingomyelin (L/S) ratio in amniotic fluid is an excellent predictor of fetal lung maturity in most pregnancies but today its value is limited. A review of the use of the L/S ratio service in Manchester from 1975 to 1981 (inclusive) shows a dramatic and consistent fall in the number of requests made over the last 18 months of this period resulting in inconvenience and inefficiency in the laboratory service. Possible explanations for the declining use of the L/S ratio service are discussed with particular reference to St Mary's Hospital, Manchester and a regional re-organization of laboratory assessment of fetal lung maturity is proposed.
Amniotic Fluid↗
Secondary structure of bovine alpha s1- and beta-casein in solution.
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Animals↗
Photodegradation of DNA with fluorescent light in the presence of riboflavin, and photoprotection by flavin triplet-state quenchers.
Superoxide anion was photogenerated upon illumination of nucleic acids with fluorescent light in a solution containing phosphate buffer, pH 7.8 and riboflavin. DNA was a better reducing substrate for this reaction than was RNA. A similar riboflavin-sensitized photoreaction caused single- and double-strand scissions of supercoiled PM2 DNA as detected by electrophoresis in agarose gels. None of specific scavengers or quenchers for superoxide anion and other active oxygen species prevented the DNA strand breaks. However, among the flavin triplet-state quenchers, potassium iodide, butylated hydroxyanisole, and ferricytochrome c protected the supercoiled DNA from photodegradation; butylated hydroxytoluene, alpha-tocopherol, tyrosine and hemoglobin did not have any protective effect. These results indicate that triplet-state riboflavin or a derivative formed from it participate directly in the observed riboflavin-sensitized DNA photodegradation and that active oxygen species are not directly involved.
Butylated Hydroxyanisole↗
14C-Methylated beta-casein as a substrate for plasmin, and its application to the study of milk protein transformations.
A method was investigated for the sensitive radiochemical assay of milk protein transformations. beta-Casein was reductively methylated with NaBH4 and H14CHO giving a product of spec. act. 4.42 microCi/mg in which a maximum of 12% of the lysine residues were labelled. Methylation did not alter the electrophoretic or chromatographic properties of the protein, or its pattern of proteolysis by plasmin. The substrate susceptibility of 14C-methylated beta-casein towards plasmin was determined by measuring radioactivity transfer to the proteolytic fragments gamma 2- and gamma 3-casein. Compared with the native protein, no decrease in substrate susceptibility was detected. The presence in milk of the plasmin-like enzyme, milk proteinase, was demonstrated and its activity at 4 degrees C was quantified by examination of the fragmentation pattern of added 14C-methylated beta-casein. It was concluded that 14C-methylated protein substrates, prepared by reductive methylation, are well suited to the study of hydrolytic enzymes and that they can provide valuable information on milk protein transformations. In particular, no interference was encountered with the rate of cleavage by plasmin when the level of methylation was kept to a minimum.
Animals↗
Levels and location of adenosine 5'-triphosphate in bovine milk.
Bovine milk systems were analysed for adenosine 5'-triphosphate (ATP) using the luciferase-ATP reaction in a liquid scintillation counter. Approximately 0.2 mumol ATP/l milk serum were evident both in whole milks and the corresponding skim-milks. ATP was not detectable in skim-milk ultrafiltrates. These findings indicated that ATP was present in a non-dialysable portion of skim-milk. Centrifugation of whole milks from individual cows at 5500 g for 15 min at 10 degrees C yielded skim-milks essentially devoid of somatic cells and bacteria. However, the ATP in the skim-milks decreased by less than 20% compared with the whole milks indicating that the calcium phosphate-citrate (CPC)--caseinate micelles were the source of the ATP. ATP was not detectable in colloidal phosphate-free milk, from which CPC had been removed, confirming that the ATP was sequestered in the constituent CPC. Likewise, the occurrence of significant amounts of Mg, another potent stabilizer of amorphous calcium phosphate (ACP) in other biological systems, was confirmed in the colloidal phosphate of milk. From 0.13 to 0.31 mumole ATP/1 (mean 0.23) was found in the 9 milk samples studied. The discovery of small but appreciable levels of ATP in the CPC of milk provides further evidence for the analogy previously shown to exist between the CPC complex of milk and the ACP which accumulates in mitochondria. The latter has been postulated to provide an essential precursor for crystalline bone salts to form in ordered calcification processes. The implications of these findings in the biosynthesis of milk are briefly discussed.
Adenosine Triphosphate↗
Affinity of phytoestrogens for estradiol-binding proteins and effect of coumestrol on growth of 7,12-dimethylbenz[a]anthracene-induced rat mammary tumors.
A number of nonsteroidal estrogens, which are common naturally occurring substances in human foods, were examined for competitive binding to estrogen receptor proteins. These compounds bound competitively to estrogen receptor proteins in rat uterine cytosol, in tissue from 7,12-dimethylbenz[a]anthracene (DMBA)-induced rat mammary tumors, and in human mammary tumor tissue. The relative affinity of these estrogens for rat uterine cytosol receptors paralleled closely the affinities reported for other receptors. Oral administration of coumestrol did not appear to support the growth of DMBA-induced rat mammary tumors, nor did coumestrol act as an antiestrogen when administered orally in combination with 17 beta-estradiol. Coumestrol administered sc might, however, be able to support the growth of these tumors.
9,10-Dimethyl-1,2-benzanthracene↗
Initiation of oxidative changes in foods.
Initiation of lipid peroxidation in foods may be accomplished by a variety of mechanisms. Two principal initiation reactions involve homolytic scission of preformed peroxides as catalyzed by metal ions and heme proteins and the reaction of activated oxygen species with the lipid substrate to yield peroxides and free radicals. Copper and cytochromes in the milk fat globule membrane may serve as focal points for initiation of lipid peroxidation by catalyzing homolytic scission of peroxides. Activated oxygen species which may be important in initiating oxidative changes in foods include singlet oxygen, hydroxyl radical, ozone, superoxide anion (perhydroxyl radical at low pH), and hydrogen peroxide. Chemical and enzymic reactions in biological materials can generate singlet oxygen, hydroxyl radical, superoxide anion, and hydrogen peroxide. Ozone is primarily a product of photoreactions in polluted air. Reactions involving singlet oxygen, hydroxyl radical, and ozone with food constituents ultimately can yield peroxides which decompose to initiate oxidative chain reactions. Superoxide anion and hydrogen peroxide are relatively inert toward organic molecules but can decompose to produce the more reactive singlet oxygen and hydroxyl radical. Inhibition of reactions initiated by reactive oxygen species in foods should be very important in preserving the oxidative stability of foods. This paper presents a brief review of possible initiation reactions for lipid peroxidation and inhibition of reactions of activated oxygen species that are of importance in food systems.
Chemical Phenomena↗
Antioxidant activity of skim milk: effect of sonication.
The antioxidant activity of skim milk was evaluated in a linoleate emulsion system with hemoglobin as a pro-oxidant. Sonication greatly increased the antioxidant activity of skim milk. The antioxidant activity of the casein fraction of milk was increased most by sonication, and this increase was nearly as great as that for skim milk, suggesting that casein was almost totally responsible for the antioxidant effect of sonication. Total sulfhydryl groups of skim milk decreased upon prolonged sonication, probably the result of the heat evolved in the process. Reactive sulfhydryl content was unchanged by sonication. Sonication had no effect on antioxidant activity of beta-lactoglobulin, reduced urease, or reduced ribonuclease, proteins with free sulfhydryl groups. Apparently sulfhydryl groups were not involved in the increased antioxidant activity of sonicated skim milk. Homogenization at 281.5 kg/cm2 did not increase the antioxidant effect of skim milk. Sonication probably disrupted casein micelles, increasing the effective concentration of casein, which could account for the increased antioxidant activity in the system.
Animals↗
Kinetics of pepsin-initiated coagulation of kappa-casein.
The kinetics of pepsin initiated coagulation of kappa-casein have been studied at pH 5.8. The primary and secondary phases of coagulation are shown to proceed simultaneously. The theory of enzymatically initiated clotting reactions proposed by Payens (Payens, T.A.J. (1976) Neth. Milk Dairy J. 30, 55--59) has been applied to this clotting system and has been used to obtain rate constants for the secondary phase of coagulation. As expected, clotting rate constants for kappa-casein increase with pepsin concentration. An activation energy of 30.6 kcal/mol has been obtained for the secondary phase of coagulation. Turbidity measurements are a convenient means for studying the secondary phase of coagulation but do not provide an unambiguous means for studying the primary phase of the reaction.
Animals↗
Applications of microbial enzymes in food systems and in biotechnology.
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Antioxidants↗
Derivatives of L-asparaginase designed to be immobilized in vivo [proceedings].
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Asparaginase↗
Absence of fatty livers in rhesus monkeys fed orotic acid.
Pairs of rhesus monkeys were fed for 10 wk a basal diet containing 1% orotic acid or 10% nonfat milk powders. Amounts of total lipids in the liver and hepatic morphology were normal after 10 wk indicating that orotic acid in the diet did not induce fatty livers in rhesus monkeys.
Animals↗