bluuebunny/crossref_metadata_2025 huggingface dataset:
Microbiological degradation of bile acids. Ring a cleavage and 7α,12α-dehydroxylation of cholic acid by Arthrobacter simplex
Authors: Hayakawa Shohei , Kanematsu Yoshiko , Fujiwara Takashi
Published: August 2015

The metabolism of cholic acid by Arthrobacter simplex was investigated. This organism effected both ring a cleavage and elimination of the hydroxyl groups at C-7 and C-12 and gave a new metabolite, (4R)-4-[4α-(2-carboxyethyl)-3aα-hexahydro-7aβ-methyl-5-oxoindan-1β-yl]valeric acid, which was isolated and identified through its partial synthesis. A degradative pathway of cholic acid into this metabolite is tentatively proposed, and the possibility that the proposed pathway could be extended to the cholic acid degradation by other microorganisms besides A. simplex is discussed. The possibility that the observed reactions in vitro could occur during the metabolism of bile acids in vivo is considered.
The isolation and characterization of trigalactosyl diglyceride from potato tubers
Authors: Galliard T.
Published: August 2015

Trigalactosyl diglyceride has been isolated from tubers of potato (Solanum tuberosum) by a combination of chromatographic methods. This galactolipid, which constitutes approximately 1% by weight of the total lipids, was characterized by analysis of the intact lipid and its deacylation product. The fatty acids:glycerol:galactose molar proportions were shown to be close to 2:1:3. Evidence was obtained that suggests that trigalactosyl diglyceride is a higher homologue of mono- and di-galactosyl diglycerides and contains an additional d-galactopyranosyl moiety that is linked α-(1→6) to the terminal galactose unit of digalactosyl diglyceride.
Radiochemical micro assays for the determination of choline acetyltransferase and acetylcholinesterase activities
Authors: Fonnum F.
Published: August 2015

1. The methods for the assay of choline acetyltransferase were based on the reaction between labelled acetyl-CoA and unlabelled choline to give labelled acetylcholine. 2. Both synthetic acetyl-CoA and acetyl-CoA formed from sodium [1−14C]acetate or sodium [3H]acetate by incubation with CoA, ATP, Mg2+ and extract from acetone-dried pigeon liver were used. 3. [1−14C]Acetylcholine was isolated by extraction with ketonic sodium tetraphenylboron. 4. [3H]Acetylcholine was precipitated with sodium tetraphenylboron to remove a ketone-soluble contaminant in sodium [3H]acetate and then extracted with ketonic sodium tetraphenylboron. 5. The values of choline acetyltransferase activity obtained in the presence of sodium cyanide or EDTA and synthetic acetyl-CoA were similar to those obtained with acetyl-CoA synthesized in situ. 6. The assay of acetylcholinesterase was based on the formation of labelled acetate from labelled acetylcholine. The labelled acetylcholine could be quantitatively removed from the acetate by extraction with ketonic sodium tetraphenylboron. 7. The methods were tested with samples from central and peripheral nervous tissues and purified enzymes. 8. The blank values for choline acetyltransferase and acetylcholinesterase corresponded to the activities in 20ng. and 5ng. of brain tissue respectively.
The relationship between nitrogen fixation and the production of HD from D2 by cell-free extracts of soya-bean nodule bacteroids
Authors: Turner G. L. , Bergersen F. J.
Published: August 2015

1. Cell-free extracts prepared from soya-bean nodule bacteroids produced HD from D2 in the presence of dithionite, an ATP-generating system and nitrogen. 2. Crude extracts of bacteroids or of Azotobacter vinelandii showed some background D2 exchange when any one of these was omitted. 3. Partial purification of bacteroid extracts diminished this background activity and gave increased D2 exchange and nitrogen fixation. 4. Although increasing pN2 stimulated both reactions, the apparent Km (N2) for nitrogen fixation was much higher than the apparent Km (N2) for D2 exchange when partially purified bacteroid extracts were used. 5. Carbon monoxide was a competitive inhibitor of nitrogen fixation by partially purified bacteroid extracts, but D2 exchange was inhibited in a non-competitive fashion. 6. These results are discussed in relation to the possible existence of enzyme-bound intermediates of nitrogen fixation.
Kinetic studies of glutamate dehydrogenase with glutamate and norvaline as substrates. Coenzyme activation and negative homotropic interactions in allosteric enzymes
Authors: Engel P. C. , Dalziel K.
Published: August 2015

1. Kinetic studies of glutamate dehydrogenase were made with wide concentration ranges of the coenzymes NAD+ and NADP+ and the substrates glutamate and norvaline. Initial-rate parameters were evaluated. 2. Deviations from Michaelis–Menten behaviour towards higher activity were observed with increasing concentrations of either coenzyme with glutamate as substrate, but not with norvaline as substrate. 3. In phosphate buffer, pH7·0, Lineweaver–Burk plots with either coenzyme as variable and a constant, large glutamate concentration showed three or four linear regions of different slope with relatively sharp discontinuities. Maximum rates obtained by extrapolation and Michaelis constants for the coenzymes increased in steps with increase of coenzyme concentration. 4. In the absence of evidence of heterogeneity of the enzyme and coenzyme preparations, the results are interpreted in terms of negative homotropic interactions between the enzyme subunits. It is suggested that sharp discontinuities in Lineweaver–Burk plots or reciprocal binding plots may be characteristic of this new type of interaction, which can be explained in terms of an Adair–Koshland model, but not by the model of Monod, Wyman & Changeux.
The nature of carbon dioxide substrate and equilibrium constant of the 6-phosphogluconate dehydrogenase reaction
Authors: Villet R. H. , Dalziel K.
Published: August 2015

1. It was shown that dissolved CO2 and not HCO3− or H2CO3 is the primary substrate for reductive carboxylation with 6-phosphogluconate dehydrogenase from sheep liver. 2. The equilibrium constant of the reaction was measured in solutions of various ionic strengths and at several temperatures, and the free energy and heat of reaction were determined.
Lipid synthesis in relation to the cell cycle of Bacillus megaterium KM and Escherichia coli
Authors: Daniels M. J.
Published: August 2015

Lipid synthesis during the cell duplication cycle of Bacillus megaterium KM and Escherichia coli was studied by glycerol incorporation both in synchronized cultures and in unsynchronized exponentially growing populations subsequently fractionated according to size (and age). A large transient increase in the rate of incorporation per unit cell mass was observed around the time of cell division, probably reflecting the synthesis of the division septum.
Oxidation of C1 compounds by Pseudomonas sp. MS
Authors: Kung Hsiang-Fu , Wagner Conrad
Published: August 2015

Pseudomonas sp. MS is capable of growth on a number of compounds containing only C1 groups. They include trimethylsulphonium salts, methylamine, dimethylamine and trimethylamine. Although formaldehyde and formate will not support growth they are rapidly oxidized by intact cells. Methanol neither supports growth nor is oxidized. A particulate fraction of the cell oxidizes methylamine to carbon dioxide in the absence of any external electron acceptor. Formaldehyde and formate are more slowly oxidized to carbon dioxide by the particulate fraction, although they do not appear to be free intermediates in the oxidation of methylamine. Soluble NAD-linked formaldehyde dehydrogenase and formate dehydrogenase are also present. The particulate methylamine oxidase is induced by growth on methylamine, dimethylamine and trimethylamine, whereas the soluble formaldehyde dehydrogenase and formate dehydrogenase are induced by trimethylsulphonium nitrate as well as the aforementioned amines.
Appendix: Mathematical approaches to the evaluation of the flux of γ-aminobutyrate in brain tissue in vitro
Authors: Hammond B. J. , Julian T. , Machiyama Y. , Balázs R.
Published: August 2015

In the preceding paper (Balázs, Machiyama, Hammond, Julian & Richter, 1970) the flux of γ-aminobutyrate (GABA) was found, in guinea-pig brain-cortex slices incubated in glucose–saline medium, to represent about 10% of the total tricarboxylic acid cycle flux, as opposed to other estimates, which are as high as 40%. However, the latter value was deduced from experimental results by methods that made no allowance for the metabolic compartmentation of glutamate: a mathematical investigation was therefore undertaken to show that this omission necessarily leads to an overestimation of GABA flux. The magnitude of this over-estimation was shown by computer simulation methods to be of such an order as to bring the corrected value into agreement with the lower value. Computer simulation methods were also used to evaluate the GABA flux from the experimental results presented by Balázs et al. (1970) and a value of 0.0315μmol/min per g wet wt. was obtained. This value was also shown to be consistent, in the simulated system, with the experimentally observed time-courses for the radioactivity and quantity of aspartate. Since there is now evidence that GABA is itself a metabolically compartmented intermediate this possibility was considered mathematically, but it was found that in this case the assumption of compartmentation had little effect upon the value of GABA flux deduced on the basis of GABA homogeneity.
Methylation of deoxyribonucleic acid in cultured mammalian cells by N-methyl-N′-nitro-N-nitrosoguanidine. The influence of cellular thiol concentrations on the extent of methylation and the 6-oxygen atom of guanine as a site of methylation
Authors: Lawley P. D. , Thatcher Carolyn J.
Published: August 2015

1. In neutral aqueous solution N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) yields salts of nitrocyanamide as u.v.-absorbing products. With cysteine, as found independently by Schulz & McCalla (1969), the principal product is 2-nitràminothiazoline-4-carboxylic acid. Both these reactions liberate the methylating species; thiols enhance the rate markedly at neutral pH values. An alternative reaction with thiols gives cystine, presumably via the unstable S-nitrosocysteine. 2. Thiols (glutathione or N-acetylcysteine) in vitro at about the concentration found in mammalian cells enhance the rate of methylation of DNA markedly over that in neutral solution. 3. Treatment of cultured mammalian cells with MNNG results in rapid methylation of nucleic acids, the extent being greater the higher the thiol content of the cells. Rodent embryo cells are more extensively methylated than mouse L-cells of the same thiol content. Cellular thiol concentrations are decreased by MNNG. Proteins are less methylated by MNNG than are nucleic acids. 4. Methylation of cells by dimethyl sulphate does not depend on cellular thiol content and protein is not less methylated than nucleic acids. Methylation by MNNG may therefore be thiol-stimulated in cells. 5. Both in vitro and in cells about 7% of the methylation of DNA by MNNG occurs at the 6-oxygen atom of guanine. The major products 7-methylguanine and 3-methyladenine are given by both MNNG and dimethyl sulphate, but dimethyl sulphate does not yield O6-methylguanine. Possible reaction mechanisms to account for this difference between these methylating agents and its possible significance as a determinant of their biological effects are discussed.
An endogeneous inhibitor of microsomal mixed-function oxidases in homogenates of the southern armyworm (Prodenia eridania)
Authors: Krieger R. I. , Wilkinson C. F.
Published: August 2015

1. A potent inhibitor of microsomal mixed-function oxidation has been isolated from the gut contents of larvae of the southern armyworm (Prodenia eridania). 2. Progressive inhibition is associated with a soluble proteinase with a molecular weight of approx. 26000. 3. Although the proteolytic activity of the inhibitor can be reduced by phenylmethanesulphonyl fluoride and other reagents, these materials are only partially effective in protecting the microsomal enzymes. 4. Inhibitory activity is decreased in the presence of bovine serum albumin.
An obstacle in preparing alloisoleucine
Authors: Laird W. M. , Matai S. , Synge R. L. M.
Published: August 2015

Alloisoleucine, prepared from a mixture of racemic isoleucine–alloisoleucine diastereoisomers, can be heavily contaminated with isoleucine if the recrystallization of acetyl-dl-alloisoleucine is controlled by melting point only. Two commercial specimens were found to be thus contaminated.
Effect of temperature acclimatization on the fatty acid composition of goldfish intestinal lipids
Authors: Kemp P. , Smith M. W.
Published: August 2015

1. The fatty acid composition of whole goldfish, whole-intestinal mucosa, intestinal mucosal membranes and individual phospholipids extracted from mucosal membranes were measured, fish adapted to different temperatures being used. 2. Alterations of the adaptation temperature did not noticeably affect the fatty acid composition of the whole-fish lipids, but there were marked changes in the fatty acids of lipids extracted from homogenates of goldfish intestinal mucosa. These changes were more pronounced in a membrane fraction prepared from these homogenates. Raising the adaptation temperature by 20°C halved the percentage of C20:1, C20:4 and C22:6 fatty acids and nearly doubled the percentage of C18:0 and C20:3 fatty acids recovered. 3. Choline phosphoglycerides constituted about one-half and ethanolamine phosphoglycerides about one-quarter of the total membrane phospholipids. 4. The fatty acids of choline and ethanolamine phosphoglycerides were more susceptible to temperature-dependent changes than were the phosphoglycerides of inositol or serine. 5. The increase in C18:0 fatty acid that occurred in membranes of warm-adapted fish was greatest for ethanolamine phosphoglycerides, but increases also occurred in other phospholipid fractions and in membrane neutral lipids.
Zymogen activation as a sensitive enzymeamplifying assay for a protease with tryptic specificity
Authors: Barrett Dennis
Published: August 2015

A protease, ‘trypsinogenase’, secreted in small amounts by the sea-urchin blastula, is assayed in two steps as an example of enzyme-amplifying kinetics. In reaction 1 the trypsinogenase catalyses the activation of trypsinogen to trypsin. In reaction 2 the trypsin catalyses the hydrolysis of N-α-toluene-p-sulphonylarginine methyl ester, at a rate that is linear with trypsinogenase concentration over a 20-fold range. Results are reproducible within a batch of zymogen, but each batch requires a separate standard curve.
Effect of cortisone on the developmental pattern of the neutral and the acid β-galactosidases of the small intestine of the rat
Authors: Koldovský Otakar , Sunshine Philip
Published: August 2015

1. The developmental pattern and effect of cortisone on acid β-galactosidase and neutral β-galactosidase were studied in postnatal rats by a recently proposed method for their independent determination. 2. After birth the acid β-galactosidase activity increases in the ileum, whereas it decreases slightly in the jejunum. On day 16 after birth the activity in the ileum decreases and in 20-day-old rats activity in both parts of the intestine decreases to adult values. In suckling animals the activity in the ileum exceeds the jejunal activity severalfold and in adult animals the activity in the jejunum is slightly higher than that in the ileum. 3. Neutral β-galactosidase activity is high after birth and decreases in both jejunum and ileum after day 20 after birth. In 12–20-day-old rats activity in both parts is essentially the same, but in adult animals jejunal activity exceeds ileal activity four-to five-fold. 4. Cortisone (0.5, 2.0 or 5.0mg/100g body wt. daily for 4 days) does not influence the activity of either enzyme in 60-day-old rats. Acid β-galactosidase activity is decreased after cortisone treatment in 8-, 12-, 16-and 18-day-old rats, with sensitivity to cortisone increasing with the approach of weaning. No effect of cortisone on acid β-galactosidase is seen in 8-day-old rats. Neutral β-galactosidase activity is increased in the ileum of 8-, 12-, 16- and 18-day old rats, but only in the jejunum of 8-and 12-day-old rats.
Enzymes involved in acetoacetate formation in various bovine tissues
Authors: Baird G. D. , Hibbitt K. G. , Lee Janice
Published: August 2015

1. The activities of acetoacetyl-CoA thiolase, hydroxymethylglutaryl-CoA synthase and lyase and acetoacetyl-CoA deacylase were measured in homogenates of samples of liver, rumen epithelium (long papillae), kidney and lactating mammary gland derived from slaughtered cows. 2. The activities of the four enzymes in bovine liver were similar to the activities previously reported for the corresponding enzymes in rat liver. 3. Acetoacetyl-CoA thiolase and hydroxymethylglutaryl-CoA synthase and lyase were present in rumen epithelium. The activities of the enzymes were all lower on a wet weight basis than in liver. Only very slight deacylase activity was detected. 4. Kidney contained acetoacetyl-CoA thiolase, hydroxymethylglutaryl-CoA lyase and acetoacetyl-CoA deacylase, but only trace amounts of hydroxymethylglutaryl-CoA synthase. 5. Mammary gland contained acetoacetyl-CoA thiolase and some hydroxymethylglutaryl-CoA lyase, but virtually no hydroxymethylglutaryl-CoA synthase or acetoacetyl-CoA deacylase. 6. Since physiologically significant ketogenesis probably occurs solely via the hydroxymethylglutaryl-CoA pathway, it is evident that, of the four tissues examined, such ketogenesis must be restricted to the liver and the rumen epithelium. 7. All the enzymes except hydroxymethylglutaryl-CoA lyase were also assayed in the four tissues derived from cows suffering from bovine lactational ketosis. Ketosis did not cause a statistically significant change in the activity of any of the enzymes measured. 8. Hepatic hydroxymethylglutaryl-CoA synthase and lyase were found to be associated mainly with the particulate fraction, as in the rat. A considerably greater proportion of these enzymes was found to be present in the cytoplasmic fraction from rumen epithelium, although it was not excluded that this was due to mitochondrial damage during homogenization. 9. Appreciable hydroxymethylglutaryl-CoA synthase was also present in epithelium from the dorsal region of the rumen, from the reticulum and from the omasum, but not from the abomasum.
Chemistry of the collagen cross-links. Isolation and characterization of two intermediate intermolecular cross-links in collagen
Authors: Bailey A. J. , Peach Catherine M. , Fowler L. J.
Published: August 2015

This paper describes the isolation from reduced collagen of two new amino acids believed to be involved, in their non-reduced form, as intermolecular cross-links stabilizing the collagen fibre. The reduction of intact collagen fibrils with tritiated sodium borohydride was found to stabilize the aldehyde-mediated cross-links to acid hydrolysis and thus allowed their location and isolation from acid hydrolysates on an automatic amino acid analyser. Comparison of the radioactive elution patterns from the autoanalyser of collagen treated in various ways before reduction permitted a preliminary classification of the peaks into cross-link precursors, intramolecular and intermolecular cross-links. The techniques employed to isolate the purified components on a large scale and to identify them structurally are described in detail. Two labile intermolecular cross-links were isolated in their reduced forms, one of which was identified by high-resolution mass spectrometry as N∈-(5-amino-5-carboxypentyl)hydroxylysine. The structure of this compound was confirmed by chemical synthesis. The cross-link precursor α-aminoadipic δ-semialdehyde was isolated in its reduced form, ∈-hydroxynorleucine, together with its acid degradation product ∈-chloronorleucine. A relatively stable intermolecular cross-link was isolated and partially characterized by mass spectrometry as an aldol resulting from the reaction of the δ-semialdehyde derived from lysine and hydroxylysine.
The inhibition of 1,1,1-trichloro-2,2-bis-(p-chlorophenyl)ethane (DDT) dehydrochlorinase and gluthathione S-aryltransferase in grass-grub and housefly preparations
Authors: Balabaskaran S. , Smith J. N.
Published: August 2015

The inhibition of DDT [1,1,1-trichloro-2,2-bis-(p-chlorophenyl)ethane] dehydrochlorinase and glutathione S-aryltransferase by diphenylmethane and triphenylmethane derivatives was examined. Bis-(3,5-dibromo-4-hydroxyphenyl)methane and similar compounds were excellent inhibitors of both enzymes, but only DDT dehydrochlorinase was inhibited by compounds similar to bis-(N-dimethylaminophenyl)methane. Colour salts of the basic triphenylmethyl dyes were excellent inhibitors of both enzymes. All the inhibitors examined appeared to act by competition with glutathione for its binding site on the two enzymes.
The fate of sulphadimethoxine in primates compared with other species
Authors: Adamson R. H. , Bridges J. W. , Kibby M. R. , Walker S. R. , Williams R. T.
Published: August 2015

1. The metabolism of sulphadimethoxine (2,4-dimethoxy-6-sulphanilamidopyrimidine) was examined in nine species of primates and nine species of non-primates. 2. The main metabolite of the drug in the urine in man, rhesus monkey, baboon, squirrel monkey, capuchin, bushbaby, slow loris and tree shrew was sulphadimethoxine N1-glucuronide. In the green monkey, although the main metabolite was N4-acetylsulphadimethoxine, the N1-glucuronide was also a major metabolite. 3. In the dog, rat, mouse, guinea pig, Indian fruit bat and hen the N1-glucuronide was a minor metabolite in the urine, whereas in the cat, ferret and rabbit this glucuronide was not found in the urine. 4. All the species examined except the dog excreted some N4-acetylsulphadimethoxine, which was the major metabolite in the green monkey, rabbit and guinea pig. 5. In the tree shrew, a doubtful primate, N1-glucuronide formation was similar to that in the other primates. 6. It is suggested that the slow excretion of the drug by the rat may be due partly to strong binding of the drug to tissue proteins and that the strength of binding may vary with species. 7. In the rat the amount of N1-glucuronide found in the urine is not a true indication of the extent of this conjugation since much more of the conjugate was found in the bile (7% of the dose) than in the urine (1%). In the rabbit, no N1-glucuronide was found in the bile or urine, but a small amount of sulphadimethoxine N4-glucuronide was found in the bile of the rat (0.5% of dose) and rabbit (0.8%).
A factor in animal tissues that causes sialyl residues of mucoproteins to react as free sialic acid in the Warren assay
Authors: Brown C. R. , Srivastava P. N. , Hartree E. F.
Published: August 2015

1. The mucin of the Cowper's gland of the boar is a sialomucoprotein similar to submaxillary-gland mucin. When a solution of either mucin has been incubated for 5min or less with a particulate fraction from homogenized uterine endometriumplus-myometrium of the rabbit, 10–20% of sialyl residues (N-acetylneuraminic acid) give a positive Warren reaction for free N-acetylneuraminic acid. The particulate fraction is devoid of neuraminidase and no free (diffusible) N-acetylneuraminic acid appears during incubation. The factor that catalyses the formation of directreading non-diffusible N-acetylneuraminic acid occurs also in liver, kidney and intestinal mucosa of the rabbit. The factor is present in very small (`microsomal') particles and has not yet been solubilized. Homogenates of boar Cowper's gland contain both factor and mucin; thus direct-reading non-diffusible N-acetylneuraminic acid appears when such homogenates are stored. 2. Under optimum conditions 1mg of uterine protein catalyses the formation of 0.05–0.1μmol of direct-reading non-diffusible N-acetylneuraminic acid/min. This activity is considerably higher than the neuraminidase activities of comparable homogenates of animal tissues or of liver lysosomes. The factor is thermostable and its activity shows little variation within (i) the pH range 3–10, (ii) the temperature range 20–37°C. Activity is inhibited strongly by 2,2′-bipyridyl and by ammonium pyrrolidine dithiocarbamate but is unaffected by EDTA. Its action can be simulated by low concentrations of Fe2+. From this it may be inferred that the factor is a protein-bound from of bivalent iron. A number of pure iron-containing proteins and haemoproteins were completely inactive. The following substrates were not sources of direct-reading non-diffusible N-acetylneuraminic acid: methoxyneuraminic acid, sialyl-lactose, brain gangliosides, and sialoproteins in which N-acetylneuraminic acid is linked to galactose residues. 3. It is proposed that the factor (or Fe2+) reacts with the mucin in a manner that renders the C-4–C-5 bond of sialyl residues susceptible to periodate oxidation.
Factors involved in changes in hepatic lipogenesis during development of the rat
Authors: Lockwood Elizabeth A. , Bailey E. , Taylor C. B.
Published: August 2015

1. Changes in the activities of acetyl-CoA carboxylase (EC 6.4.1.2), phosphofructokinase (EC 2.7.1.11), aldolase (EC 4.1.2.13), extramitochondrial aconitate hydratase (EC 4.2.1.3) and NADP-dependent isocitrate dehydrogenase (EC 1.1.1.42) have been measured in the livers of developing rats from late foetal life to maturity. 2. The effect of altering the weaning time on some enzymes associated with lipogenesis has been studied. Weaning rats at 15 days of age instead of 21 days results in an immediate increase in the activity of `malic' enzyme (EC 1.1.1.40) whereas the activities of glucose 6-phosphate dehydrogenase (EC 1.1.1.49) and ATP citrate lyase (EC 4.1.3.8) did not increase until 4–5 days and acetyl-CoA carboxylase 2–3 days after early weaning. Weaning rats on to an artificial-milk diet led to complete repression of the rise in activity of hepatic enzymes associated with lipogenesis normally found on weaning, except for `malic' enzyme, which increased in activity after 20 days of age. 3. The effect of intraperitoneal injections of glucagon, cortisol, growth hormone and thyroxine on the same hepatic enzymes has been investigated. Only thyroxine had any effect on enzyme activities and caused a 20-fold increase in `malic' enzyme activity and a twofold increase in ATP citrate lyase activity. 4. The activities of hepatic glucose 6-phosphate dehydrogenase and `malic' enzyme are higher in adult female than in adult male rats and it has been shown that this sex difference in enzyme activities is due to both male and female sex hormones. 5. Hepatic malate, citrate, pyruvate, glucose 6-phosphate and phosphoenolpyruvate concentrations have been measured throughout development. 6. The results are discussed in relation to the dietary and hormonal control of hepatic enzyme activities during development.
Galactothermin, a reversibly heat-precipitable protein of human milk at neutral pH
Authors: Schade A. L. , Reinhart R. W.
Published: August 2015

1. A protein, aggregating at body temperature and solubilizing when cooled, was isolated from fresh human milk at neutral pH and studied for some of its physical, chemical and immunological properties. The name `galactothermin' is proposed for this protein. 2. Isolation and purification of galactothermin involved casein removal from skim milk at pH4.64 followed by centrifugal fractionation of residual protein-containing solutions repeatedly heated and cooled between 40°C and 0°C at pH7.3. 3. The molecular weight by ultracentrifugal analysis and the minimum molecular weight by sum of amino acid residues were 11400 and 14000 respectively. The sedimentation coefficient s25,w was 1.05S and the diffusion coefficient was 7.15×10−7. Reversible aggregation is favoured by increase in protein concentration, ionic strength, temperature, time and approach to the isoionic point of 7.27 from either acidic or alkaline conditions. 4. Among the amino acid residues, proline predominates and non-polar species account for two-thirds of the total. Cysteine and cystine are absent. Analysis of galactothermin showed it to be essentially free of hexose, sialic acid, calcium and phosphate. 5. Galactothermin is antigenic in the rabbit as evidenced by the passive cutaneous anaphylaxis test. Single precipitin lines are produced in immunodiffusion tests. 6. By electrophoresis in polyacrylamide gel at pH4.0 only one sharp band is produced.
Lipid metabolism in the perfused chicken liver. Lipogenesis from glucose, acetate and palmitate
Authors: Bickerstaffe R. , West C. E. , Annison E. F.
Published: August 2015

A procedure for the perfusion of the isolated chicken liver was developed. The preparation satisfied many of the criteria of normal physiological function, e.g. oxygen consumption and the absence in the perfusion medium of enzymes indicative of cell damage, and retained its capacity to synthesize lipids from glucose, acetate and long-chain fatty acids. Part of newly synthesized triglyceride was released into the perfusion medium as lipoprotein.
A study on the tricarboxylic acid cycle and the synthesis of acetylcholine in the lobster nerve
Authors: Cheng S.-C. , Nakamura R.
Published: August 2015

1. The pattern of metabolism of 14C-labelled substrates in the lobster nerve suggested a normal tricarboxylic acid cycle with a slow turnover. 2. Acetylcholine was synthesized from [2-14C]acetate, [2-14C]pyruvate and [1,5-14C]citrate, implying the presence of acetate thiokinase, choline acetylase and citrate-cleavage enzyme. 3. [2-14C]Acetate was the best precursor. 4. The formation of acetyl-CoA from citrate was limited, probably by the citrate-cleavage enzyme, although the magnitude of the reversed reactions of the tricarboxylic acid cycle was large when compared with that of the forward reactions. 5. The relative magnitude of the two pathways (acetyl-CoA and carbon dioxide fixation) in pyruvate utilization was nearly equal. 6. The probable presence of metabolic compartments in the lobster nerve is discussed.
The composition of β-lactamase I and β-lactamase II from Bacillus cereus 569/H
Authors: Kuwabara S. , Adams E. P. , Abraham E. P.
Published: August 2015

1. Crystalline β-lactamase I from Bacillus cereus 569/H yielded only amino acids on acid hydrolysis, but crystalline β-lactamase II from the same organism yielded also substantial quantities of neutral sugars and amino sugars. 2. Analysis with an amino acid analyser indicated that the two enzymes were similar though not identical in overall amino acid composition. Analysis of neutral and amino sugars as their silyl derivatives by gas–liquid chromatography showed that the carbohydrate moiety of β-lactamase II contained residues of glucose, galactose, mannose, fucose, glucosamine and galactosamine. 3. After oxidation and hydrolysis both β-lactamases gave small amounts of cysteic acid. After treatment of inactive Zn2+-free β-lactamase II with N-ethylmaleimide or iodoacetate enzymic activity was not restored by the addition of Zn2+.
Acid-soluble phosphorus compounds in mammalian semen
Authors: Brooks D. E.
Published: August 2015

1. A method is described for the extraction, purification and separation of acid-soluble phosphorus compounds from mammalian semen. [8-14C]ATP and [8-14C]AMP were used as internal recovery standards to measure the breakdown and loss of these nucleotides in the procedure. 2. Bull, ram, boar and stallion semen was separated into seminal plasma and spermatozoa and the two fractions were examined separately. The overall composition of the mixture of the phosphorus compounds extracted from the two fractions was similar for the four species. 3. Glycerylphosphorylcholine and glycerylphosphorylinositol were the two phosphorus compounds identified in extracts of seminal plasma. ATP, ADP, AMP, GTP, GDP, NAD, fructose 1,6-diphosphate and glucose 6-phosphate were identified in extracts prepared from spermatozoa.
Metabolism of the cellular slime mould Dictyostelium discoideum grown in axenic culture
Authors: Ashworth J. M. , Watts D. J.
Published: August 2015

1. The DNA, RNA, protein and carbohydrate contents of myxamoebae of Dictyostelium discoideum strain Ax-2 were measured after growth on bacteria or in various axenic media. 2. Myxamoebae grown in the different axenic media have similar DNA, RNA and protein contents, but there are marked differences in the contents of glycogen and free sugars. The DNA and protein contents of myxamoebae grown on bacteria are different from those in myxamoebae grown axenically. 3. Approximately half the DNA found in myxamoebae grown on bacteria is of bacterial rather than of slime-mould origin. 4. The specific activities of some enzymes (including UDP-glucose pyrophosphorylase) are higher in myxamoebae grown axenically than in myxamoebae grown on bacteria. Nevertheless the characteristic increase in the specific activity of UDP-glucose pyrophosphorylase occurring during differentiation of cells of the wild-type strain NC-4 is also found in cells grown axenically. 5. The rate of amino acid oxidation during axenic growth of the myxamoebae is decreased when the cells are supplied with glucose.
Biological activity and the 3-methylhistidine content of actin and myosin
Authors: Johnson P. , Perry S. V.
Published: August 2015

1. The 3-methylhistidine content of myosin varies according to muscle type. It is highest in myosin from white skeletal muscle and lower values are obtained from myosin of red skeletal and smooth muscle. 2. The 3-methylhistidine content of actin was similar in all of the types of muscle from which it was isolated. 3. The 3-methylhistidine of rabbit actin is localized in a single tryptic peptide that was readily modified during fractionation procedures. 4. Photo-oxidation studies indicated that the 3-methylhistidine residues are not essential for adeonsine triphosphatase and actin-combining activities of myosin. 5. During photooxidation G-actin lost completely the ability to polymerize to the F form before all the 3-methylhistidine was destroyed.
The cerebral sodium-plus-potassium ion-stimulated adenosine triphosphatase of bovine brain and its microsomal matrix
Authors: Pull I. , McIlwain H.
Published: August 2015

1. Adenosine triphosphatase activities of dispersions prepared from bovine cerebral cortex that had been frozen, were greater than those of dispersions prepared from fresh tissue. The subcellular distribution of components of the dispersion was not altered by freezing the tissue and a microsomal fraction enriched in Na++K+-stimulated adenosine triphosphatase activity was prepared. 2. The bovine cerebral microsomes were further treated with a 2m-sodium iodide reagent to obtain a particulate preparation with minimal Na++K+-independent adenosine triphosphatase activity. Na++K+-stimulated activity was increased by the sodium iodide treatment and this preparation was shown to be enriched in lipid constituents. 3. Density-gradient centrifugation of the sodium iodide treated preparation gave three main subfractions each containing approximately equal amounts of phospholipid and protein. Further exposure of the sodium iodide-treated preparation to the 2m-sodium iodide reagent altered the distribution of protein and phospholipid among the fractions obtained by density-gradient centrifugation. Dissociation of phospholipids from protein in the sodium iodide-treated preparation was brought about also by high concentrations of arginine. Concentrated solutions of arginine and sodium thiocyanate brought about dissociation of phospholipids from protein of the microsomal preparation. 4. Many amino acids were found to inhibit Na++K+-stimulated adenosine triphosphatase activity when present in high concentrations. The inhibition was complex but resulted, in part at least, from diminished affinity for ATP and Na+ in the presence of the amino acids. 5. A non-ionic detergent, Lubrol W, solubilized up to 40% of the enzyme activity of the sodium iodide-treated preparation together with 30% of the protein and phospholipid in the preparation. Protein was released from the sodium iodide-treated preparation by pancreatic elastase but Na++K+-stimulated adenosine triphosphatase activity of the residue was diminished. Ultrasonic treatment of the sodium iodide-treated preparation failed to release a significant proportion of Na++K+-stimulated adenosine triphosphatase activity into a form not deposited by ultracentrifugation.
The localization of some coenzyme A-dependant enzymes in rat liver mitochondria
Authors: Haddock B. A. , Yates D. W. , Garland P. B.
Published: August 2015

1. CoA, acetyl-CoA, l-carnitine and acetyl-l-carnitine when added to rat liver mitochondria equilibrate with approximately two-thirds of the total intramitochondrial water. The mitochondrial space calculated to be freely permeable to these solutes was identical with that obtained for sucrose. 2. Acetyl-CoA is rapidly deacylated by rat liver mitochondria at 0°C, and special precautions are required to measure its mitochondrial permeation. 3. Rat liver mitochondria were separated into fractions that correspond to the inner membrane, the outer membrane, and the soluble proteins of the matrix and intermembrane compartment. Soluble enzymes considered to be located in the matrix were citrate synthase (EC 4.1.3.7), palmitoyl-CoA dehydrogenase (EC 1.3.2.2), electron-transferring flavoprotein, medium-chain-length ATP-specific fatty acyl-CoA synthetase (EC 6.2.1.2), l-3-hydroxybutyryl-CoA dehydrogenase (EC 1.1.1.35) and 3-keto-acyl-CoA thiolase (EC 2.3.1.16). Carnitine palmitoyltransferase (EC 2.3.1.–) is largely associated with the inner-membrane fraction. A long-chain-length ATP-specific fatty acyl-CoA synthetase (EC 6.2.1.3) is associated with the outer-membrane fraction.
The effect of hypermethylation on the functional properties of transfer ribonucleic acid. Ribosome-binding and polypeptide synthesis
Authors: Hay J. , Pillinger D. J. , Borek E.
Published: August 2015

1. Phenylalanyl-tRNA formed after chemical hypermethylation of Escherichia coli B tRNA was able to bind to ribosomes with the same efficiency as normal phenylalanyl-tRNA. 2. Under incubation conditions used in the ribosome-binding assay, hypermethylation of tRNA did not measurably decrease the stability of either inter-nucleotide phosphodiester bonds or the covalent bond between amino acid and tRNA in phenylalanyl-tRNA. 3. The ability of hypermethylated tRNA to take part in polyphenylalanine synthesis was inhibited progressively as the degree of hypermethylation increased. 4. Hypermethylation of tRNA affected polyphenylalanine synthesis at the stage of amino acid recognition and at a further point in the synthesis but not at the level of codon–anticodon recognition. 5. The formation of polylysine was more seriously affected by hypermethylation of tRNA than would be accounted for by inhibition of amino acid acceptance alone. 6. Polyproline formation was completely inhibited by the presence of 7mol% excess of methyl groups in tRNA. 7. The possibility of a link between amino acid acceptance and ribosome-binding was suggested for phenylalanyl-tRNA, but not for lysyl- or prolyl-tRNA.
Amino acid requirement for the growth-hormone stimulation of incorporation of precursors into protein and nucleic acids of liver slices
Authors: Clemens M. J. , Korner A.
Published: August 2015

1. Incorporation of [14C]leucine into protein in rat liver slices, incubated in vitro, increased as the concentration of unlabelled amino acids in the incubation medium was raised. A plateau of incorporation was reached when the amino acid concentration was 6 times that present in rat plasma. Labelling of RNA by [3H]orotic acid was not stimulated by increased amino acid concentration in the incubation medium. 2. When amino acids were absent from the medium, or present at the normal plasma concentrations, no effect of added growth hormone on labelling of protein or RNA by precursor was observed. 3. When amino acids were present in the medium at 6 times the normal plasma concentrations addition of growth hormone stimulated incorporation of the appropriate labelled precursor into protein of liver slices from normal rats by 31%, and into RNA by 22%. A significant effect was seen at a hormone concentration as low as 10ng/ml. 4. Under the same conditions addition of growth hormone also stimulated protein labelling in liver slices from hypophysectomized rats. Tissue from hypophysectomized rats previously treated with growth hormone did not respond to growth hormone in vitro. 5. No effect of the hormone on the rate or extent of uptake of radioactive precursors into acid-soluble pools was found. 6. Cycloheximide completely abolished the hormone-induced increment in labelling of both RNA and protein. 7. It was concluded that, in the presence of an abundant amino acid supply, growth hormone can stimulate the synthesis of protein in rat liver slices by a mechanism that is more sensitive to cycloheximide than is the basal protein synthesis. The stimulation of RNA labelling observed in the presence of growth hormone may be a secondary consequence of the hormonal effect on protein synthesis. 8. The mechanism of action of growth hormone on liver protein synthesis in vitro was concluded to be similar to its mechanism of action in vivo.
Conversion of 5-(1,2-epoxy-2,6,6-trimethylcyclohexyl)-3-methylpenta-cis-2-trans-4-dienoic acid into abscisic acid in plants
Authors: Milborrow B. V. , Noddle R. C.
Published: August 2015

(±)-5-(1,2-Epoxy-2,6,6-trimethylcyclohexyl) -3-methyl[2-14C]penta-cis-2-trans-4-dienoic acid is converted into abscisic acid by tomato fruit in 1.8% yield (or 3.6% of one enantiomer if only one is utilized) and 15% of the abscisic acid is derived from the precursor. The 2-trans-isomer is not converted. The amounts of [2-3H]mevalonate incorporated into abscisic acid have shown that the 40-times higher concentration of (+)-abscisic acid in wilted wheat leaves in comparison with unwilted ones reported by Wright & Hiron (1969) arises by synthesis. The conversion of (±)-5-(1,2-epoxy-2,6,6-trimethylcyclohexyl) -3-methyl-[2-14C]penta-cis-2-trans-4-dienoic acid into abscisic acid by wheat leaves is also affected in the same way by wilting and it is concluded from this that the epoxide or a closely related compound derived from it is on the biosynthetic pathway leading to abscisic acid. The oxygen of the epoxy group was shown, by 18O-labelling, to become the oxygen of the tertiary hydroxyl group of abscisic acid.
Water uptake and energy metabolism in brain slices from the rat
Authors: Okamoto K. , Quastel J. H.
Published: August 2015

1. Exchange of 3H2O with H2O takes place rapidly in incubated rat brain slices but at a lower rate in slices from infant brain than from adult brain. The temperature coefficient (Q10) of the exchange process, between 37 and 4°C, is 1.76 with infant brain and 1.26 with adult brain. The exchange process is unaffected by the presence of ouabain or 2,4-dinitrophenol. 2. An approximately linear relationship exists between water uptake and the concentration of ATP in the incubated slices in the presence of various concentrations of glucose. Little or no change occurs in water uptake and ATP concentration in the presence of a glucose concentration exceeding 3mm. A linear relationship also exists between the water uptake and ATP concentration in the presence of 10mm-glucose and various concentrations of sodium l-glutamate but the line is parallel to that found with changed glucose concentrations and shifted in the direction of increased water uptake. A similar parallel relationship exists between water uptake and ATP concentration in the presence of 2,4-dinitrophenol, but the amount of water uptake is significantly smaller in the presence of 2,4-dinitrophenol than in its absence. 3. Copper chloride (0.3mm) or mercuric chloride (0.3mm) both increase water uptake and diminish the ATP concentration in slices. Sodium malonate (2mm) or sodium d-glutamate (10mm) has similar effects. 4. Substances, or conditions, affecting water uptake in incubated brain slices may be divided roughly into two classes in accordance with their effects on adenosine triphosphatase and membrane permeability, but there may be considerable lack of specificity.
The effect of rotor velocity on sedimentation coefficients
Authors: Charlwood P. A.
Published: August 2015

Sedimentation-coefficient measurements on human IgG, fibrinogen and α2-macroglobulin and pig thyroglobulin were made at rotor velocities of 15220–59780 rev./min and 25.0°C in capillary-type synthetic-boundary cells or ordinary cells. At the lowest velocity, IgG and fibrinogen gave results several per cent higher than at other velocities, whereas α2-macroglobulin and thyroglobulin gave values only about 1.5% higher. This behaviour of IgG and fibrinogen is attributed primarily to imperfect initial boundaries.
Factors influencing the activity of succinate dehydrogenase in membrane preparations from Micrococcus lysodeikticus
Authors: Owen Peter , Freer John H.
Published: August 2015

1. Some properties of succinate dehydrogenase [succinate–(acceptor) oxidoreductase, EC 1.3.99.1] in membrane preparations from Micrococcus lysodeikticus (N.C.T.C. 2665) were investigated. 2. In the spectrophotometric assay system adopted the reaction velocity was shown to be proportional to the amount of membrane added. Dichlorophenol-indophenol, reduced photochemically in the presence of phenazine methosulphate, or enzymically by the membrane-bound enzyme, was shown to undergo reoxidation in the dark. 3. The membrane-bound enzyme was found to be inactivated at temperatures above 10°C. 4. The specific activity of membrane-bound succinate dehydrogenase was found to increase between two- and three-fold in diluted membrane preparations equilibrated at 0°C for 6h. Membranes treated with sodium deoxycholate showed no enzyme activation on dilution but displayed maximal activity, all activity being sedimentable at 103000g. The increase in specific activity observed on dilution could be partially inhibited by fixation with glutaraldehyde, or by the presence of bovine serum albumin. 5. The addition of Mg2+ or Ca2+ ions to membrane suspensions caused an overall depression of enzyme activity. 6. The results suggest the presence of an `inhibitor' that affects the expression of membrane bound succinate dehydrogenase activity.
Identification of the acrylate porphyrin S-411 from meconium
Authors: French J. , Nicholson D. C. , Rimington C.
Published: August 2015

A new porphyrin isolated from calf and foal meconium has been characterized as a tris-(2-carboxyethyl)-mono-(2-carboxyvinyl)-tetramethyl porphin, that is as an analogue of coproporphyrin in which one propionate group has been replaced by an acrylate group. The porphyrin is mainly a type III isomer and its possible significance in haem biosynthesis is discussed.
N-(5′-Phosphoribosyl)anthranilate isomerase–indol-3-ylglycerol phosphate synthetase of tryptophan biosynthesis. Relationship between the two activities of the enzyme from Escherichia coli
Authors: Creighton Thomas E.
Published: August 2015

Further evidence is presented to confirm the previous conclusion that the enzyme from Escherichia coli catalysing the two sequential reactions in tryptophan biosynthesis, N-(5′-phosphoribosyl)anthranilic acid (PRA) → 1-(o-carboxyphenyl-amino)-1-deoxyribulose 5-phosphate (CdRP) → indol-3-ylglycerol phosphate (InGP)+CO2+H2O, consists of a single polypeptide chain. The kinetic properties of the enzyme demonstrate that intermediate CdRP formed from PRA must dissociate from the enzyme before it can be converted into InGP. It is concluded that there are two distinct and non-overlapping catalytic sites on the enzyme for the two reactions. The expected complementation between a mutationally altered form of the enzyme lacking the first reaction and a mutationally altered form lacking the second reaction has been demonstrated in vitro by InGP formation from PRA. This system thus exhibits intracistronic complementation with a non-oligomeric protein gene product.