bluuebunny/crossref_metadata_2025 huggingface dataset:
NUTRITIONAL EVALUATION OF AN ENSILED PRODUCT FROM FISH
Authors: Krishnaswamy M. A. , Kadkol S. B. , Revankar G. D.
Published: December 2009

Ensiled fish was prepared from a local variety of freshwater fish (Barbus carnaticus) by fermentation with a pure culture of Streptococcus lactis, commercial lactose being used as a source of fermentable carbohydrate. The fermented material (pH 4.7) was roller dried. The finished product was cream colored and had a somewhat aromatic odor. It had a protein content of about 72%. Total lysine, available lysine, methionine, cystine, and tryptophan of the ensiled fish (expressed as g/16 g N) were 10.1, 8.1, 3.6, 1.1, and 1.2%, respectively. Hygienically, the product, being free from coliforms, enterococci, Salmonella, coagulase-positive staphylococci, and pathogenic anaerobes, was satisfactory. The biological value of the product as determined by protein efficiency ratio (3.3), net protein utilization (82.3%), and net protein ratio (4.2) was not significantly different from that of skim milk powder, which has a protein efficiency ratio of 3.2, net protein utilization of 82.8%, and net protein ratio of 4.9.
LYTIC ENZYMES OF SORANGIUM SP.: ISOLATION AND ENZYMATIC PROPERTIES OF THE α- AND β-LYTIC PROTEASES
Authors: Whitaker D. R.
Published: December 2009

Procedures are described for the isolation of two lytic enzymes from culture filtrates of a species of Sorangium. The enzymes, designated α-lytic protease and β-lytic protease, are responsible for most of the filtrate's lytic activity towards Arthrobacter globiformis cells. The enzymes were adsorbed from the filtrate by Amberlite CG50, separated by displacement from the resin with citrate buffer containing a gradient of sodium citrate concentration, and refractionated on columns of the same resin. Trace impurities in the β-enzyme were removed by precipitation of the enzyme with ammonium sulfate. The β-enzyme has been crystallized.On electrophoresis in Tris buffer of pH 8.0, the α-enzyme migrates slightly faster than egg-white lysozyme, the β-enzyme slightly slower. The absorptivity of the α-enzyme at 280 mμ was estimated to be 0.89; that of the β-enzyme 2.05.Low concentrations of the β-enzyme lyse suspensions of Arthrobacter globiformis cells completely, and moderately higher concentrations lyse suspensions of Micrococcus lysodeikticus cells completely; corresponding concentrations of the α-enzyme lyse the suspensions incompletely. The concomitant changes in A660of the suspensions are consistent with zero-order kinetics for the β-enzyme and first-order kinetics for the α-enzyme. Untreated and partially lysed suspensions of Arthrobacter cells show little difference in the dependence of their absorbances on wavelength; corresponding suspensions of Micrococcus cells show marked differences in this respect, and the nature of the change suggests that the breakup of clumps of cells is responsible for a substantial part of the change in absorbance measured during lysis of suspensions of Micrococcus cells. Phase-contrast photomicrographs of cells undergoing lysis show that individual cells vary greatly in their rates of lysis; swelling and decreases in the refractive index precede fragmentation of the cell and may contribute appreciably to the change in absorbance of the suspension.Both enzymes hydrolyze casein. The α-enzyme has the greater activity towards this substrate.
EFFECT OF DIETARY NITRITE ON THE LIVER STORAGE OF VITAMIN A IN THE RAT
Authors: Phillips W. E. J.
Published: December 2009

When rats were fed 1% potassium nitrite in diets containing carotene or vitamin A, their livers contained less vitamin A than those of control rats. After prolonged feeding of potassium nitrite the total amount of vitamin A stored in the liver after a single dose of carotene or vitamin A was reduced, whereas there was no reduction after the administration of vitamin A aldehyde. Ingestion of nitrite did not affect the ability of rate to store vitamin A in the liver nor alter the rate of disappearance of the vitamin from the liver. It is suggested that dietary nitrite degrades carotene and vitamin A in the digestive tract before their absorption, and does not affect the conversion of carotene, the transport of the vitamin, nor the function of the liver.
ACCUMULATION OF GUANINE NUCLEOTIDES IN THE ACID-SOLUBLE FRACTION OF BACILLUS SUBTILIS INHIBITED BY ACTINOMYCIN D
Authors: Jacoli G. G. , Zbarsky S. H.
Published: December 2009

Acid-soluble extracts were made of cells of Bacillus subtilis grown on a medium containing glycine-2-14C as a labeled precursor. Paper chromatography of the extracts with propan-2-ol:H2O:NH3as solvent and radioautography of the chromatograms showed that if growth of the cells was inhibited by including actinomycin D in the medium, two radioactive ultraviolet-absorbing components appeared close to the origin of the chromatograms. Only traces were present in control cells not inhibited by the antibiotic. Spectrophotometry of eluted material indicated that these were compounds of guanine, and electrophoresis and phosphorus determinations provided evidence that they were guanosine triphosphate and guanosine diphosphate. Further evidence for this was obtained y comparing them with authentic nucleotides by thin-layer chromatography and by showing that treatment with phosphomonoesterase liberated guanosine. The presence of other components in the acid-soluble fractions was observed and two have been identified tentatively. One may be cytidine diphosphate, which is more strongly radioactive in the control cells. The second, which appears to be radioactive inosinic acid, was present in the inhibited cells but not in the control material.
A NUTRITIONAL COMPARISON OF RAPESEED OIL AND SOYBEAN OIL
Authors: Alexander J. C. , Mattson F. H.
Published: December 2009

Rapeseed oil was fed to rats at a level as high as 73% of the calories (48% by weight) of the diet, and there was no mortality in 7 weeks. This is in contrast to a report by an earlier investigator that there was complete mortality in an average of 17 days. The animals survived because the oil was gelatinized with ethyl cellulose before it was mixed into the diet, to prevent settling of the solid ingredients. When the fat levels were high, animals on the diets with rapeseed oil consumed less food and grew less than those on comparable diets with soybean oil. On the basis of caloric efficiency and ability to support growth, however, rapeseed oil was essentially equivalent to soybean oil. Thus the earlier report of the toxicity of rapeseed oil is attributable to the unpalatable form in which it was fed, and not to the oil itself. Levels of cholesterol in the blood and liver were similar whether the rats consumed diets containing rapeseed oil or soybean oil, whereas the adrenal cholesterol was increased when rapeseed oil was fed.
OVALBUMIN AND OTHER WATER-SOLUBLE PROTEINS IN AVIAN YOLK DURING EMBRYOGENESIS
Authors: Saito Z. , Martin W. G.
Published: December 2009

The proportion of water-soluble protein in egg yolk increased markedly after 12 days' incubation, concurrent with the appearance of a new fraction that evidently entered from the egg white. The electrophoretic, sedimentation [Formula: see text], molecular weight (45 × 103), and chromatographic properties of this new fraction, after 18 days' incubation, were the same as, and the tyrosine–tryptophane and phosphorus–nitrogen ratios were similar to those of authentic ovalbumin. Ovalbumin is known to have two components differing in phosphorus content and both were evident in the authentic sample and in the new yolk fraction. At the later stages of incubation the ovalbumin component of lowest phosphorus content predominated and this also may be the form that was present in the serum of the embryo. Changes in the other components of the water-soluble material during embryogenesis were less dramatic. Chromatography on DEAE-cellulose revealed at least nine yolk components in unincubated egg, although only the three livetins were made evident by free-boundary electrophoresis. The great increase in ovalbumin prevented detection of changes in the livetins during embryogenesis but α-livetin (serum albumin) apparently disappeared most rapidly and completely.
PYROCATECHOLPHTHALEIN AS A SUBSTRATE FOR CATECHOLO-METHYLTRANSFERASE
Authors: Anderson P. J. , D'Iorio A.
Published: December 2009

The incubation of pyrocatecholphthalein with S-adenosylmethionine in the presence of catechol O-methyltransferase produces a compound which can be colored by the addition of borate buffer, pH 10, and read at 595 mμ. The increase in color intensity is linear in relation to both time and enzyme concentration.Thin-layer chromatography indicates the production of two colored substances during incubation. If S-adenosylmethionine-methyl-14C is used as methyl donor, it can be shown that the colored substances are also labeled with methyl-14C. Synthetically prepared guaiacolphthalein migrates in the same way and exhibits the same visible spectrum. It is believed that the second spot is the phthalein derivative with one catechol and one guaiacol ring.The enzymatic methylation of catecholphthalein can be used as a rapid method for the determination of catechol O-methyltransferase.
ACETATE METABOLISM IN MYCOBACTERIA
Authors: Parvin R. , Pande S. V. , Venkitasubramanian T. A.
Published: December 2009

Acetate-1-14C was added to the culture medium of different species of mycobacteria in the exponential phase of growth. Analyses after incubation for 2 hours showed that acetate was utilized for the synthesis of various lipid fractions, and also converted into carbon dioxide. Mycobacterium tuberculosis strains, in contrast to saprophytic mycobacteria, showed preferential utilization of acetate for oxidation over utilization for lipid synthesis. In the virulent strain of M. tuberculosis a greater proportion of the total radioactivity in lipids was present in the non-saponifiable fraction than in other mycobacteria, including M. tuberculosis H37Ra.
THE EFFICACY OF FRAGMENTED IMMUNE SERUM GLOBULIN IN PASSIVE IMMUNIZATION
Authors: Painter R. H. , Walcraft M. J. , Weber J. C. W.
Published: December 2009

The efficacy of an aged, fragmented preparation of immune serum globulin (human) as an agent for passive immunization was examined by comparing its performance in passive immunization with that of a fresh, unfragmented preparation. Guinea pigs passively immunized with the fresh preparation were protected against doses of tetanus toxin that were lethal to pigs which had received similar amounts of tetanus antitoxin as aged material.Monkeys passively immunized by intramuscular or intravenous injections of fresh immune serum globulin had levels of polio antibody in their serum which were consistently seven times higher than those produced by injection of the aged material. Urine collected from the monkeys treated with the aged material contained quantities of both polio and tetanus antibody as Fab fragment. After the intravenous injection of the aged material more than half of the injected antibody appeared in the urine within 24 hours.
LYTIC ENZYMES IN THE DIGESTIVE JUICE OF HELIX POMATIA: CHITINASES AND MURAMIDASES
Authors: Takeda H. , Strasdine G. A. , Whitaker D. R. , Roy C.
Published: December 2009

Two enzymes in the digestive juice of Helix pomatia are responsible for most of the juice's lytic activity towards Micrococcus lysodeikticus. The enzymes were isolated by gel filtration in polyacrylamide (Biogel P-30) followed by gradient displacement from Amberlite CG50. Both enzymes hydrolyze mucopeptide from the cell wall of M. lysodeikticus and the hydrolysis is accompanied by liberation of soluble reducing sugar. The reducing end groups were shown to be N-acetyl-muramic acid residues by evidence that the hexitol of muramic acid is the only amino sugar hexitol which is released from the mucopeptide by the reaction sequence: enzymic hydrolysis, reduction with sodium borohydride, and acid hydrolysis. Thus both enzymes are muramidases. One enzyme (designated muramidase I), like egg-white lysozyme, readily hydrolyzes hydroxyethyl chitin; the other enzyme (muramidase II) does not. Muramidase II is the more basic of the two enzymes; its isoelectric point in 0.05 M hydrochloric acid – Tris buffer is close to pH 9.0. The molecular weights of muramidase I and muramidase II were estimated by the Archibald method to be approximately 21,000 and 24,000 respectively; both estimates assume a partial specific volume of 0.72 cc/g.
BIOSYNTHESIS OF OOSPOREIN IN BEAUVERIA BASSIANA (BALS.) VUILL
Authors: El Basyouni Sohair H. , Vining L. C.
Published: December 2009

Substrates labeled with carbon-14 were used to investigate the biosynthesis of oosporein in cultures of Beauveria bassiana. Acetate-1-14C and acetate-2-14C were good precursors. Degradation of the pigment by oxidation with chromic acid gave acetic acid which was labeled in the carboxyl and methyl carbon atoms, respectively, with little randomization. It also indicated that six labeled atoms of the first substrate and eight of the second had been incorporated. DL-Mevalonic acid-2-14C, shikimic acid-U-14C, and L-methionine-methyl-14C were not incorporated. Formate-14C was incorporated but did not label the methyl substituents preferentially. Short-term feeding experiments provided evidence that both acetate and malonate units participate in the biosynthesis of oosporein. Orsellinic acid-2-14C, its ethyl ester, and orcinol-3-14C were incorporated into oosporein without randomization of the label.
THE ISOLATION AND CHARACTERIZATION OF AN ACID-SOLUBLE PROTEIN FROM MYELIN
Authors: Lowden J. A. , Moscarello M. A. , Morecki R.
Published: December 2009

Myelin was isolated from central white matter of beef and human brains. It contained less than 0.1% ribonucleic acid, less than 0.03% deoxyribonucleic acid, and no glycolipid N-acetylneurammic acid. The preparation was 96% soluble in chloroform–methanol. Most of the insoluble material was sucrose. The myelin contained 19.6% protein, 17.4% cholesterol, 36.1% phospholipid, and 23.6% galactolipid. A protein extracted from the myelin preparation with cold 0.2 N sulfuric acid gave a single sedimenting boundary in the Spinco model E analytical ultracentrifuge. In 8 M urea–formate starch gels (pH 3.4) it migrated as a single band toward the cathode. Amino acid analysis revealed that the protein contained more than 20% dibasic amino acids, but little methionine and no cystine. It was rapidly hydrolyzed by trypsin. The molecular weight was estimated at 10,000. When injected intradermally into guinea pigs, in suspension in Freund's adjuvant, it caused weight loss, a flaccid paralysis, occasional convulsions, and death, a pattern characteristic of experimental allergic encephalomyelitis. Evidence is cited for the presence of at least one other protein in purified myelin.
THE ROLE OF A PROTEASE IN SPORULATION OF PENICILLIUM JANTHINELLUM
Authors: Thangamani A. , Hofmann T.
Published: December 2009

Although peptidase A, the trypsinogen-activating acid protease of Penicillium janthinellum, is formed during the post-logarithmic phase of growth and while the organism is actively sporulating, there is no direct correlation between the production of the enzyme and sporulation. The evidence for this is threefold: (a) in surface cultures acridine orange almost completely suppresses enzyme formation but reduces sporulation to a much smaller extent; (b) 6-ethyl-thiopurine decreases spore formation but stimulates the production of peptidase A; (c) transfer of mycelium in the log phase from a complete, synthetic medium to a medium lacking nitrate induces the formation of high enzyme levels without affecting the spore count.The possibility of an indirect connection between sporulation and peptidase production is not ruled out.
THE INFLUENCE OF COCAINE ON THE METABOLISM OF ADRENALINE IN VIVO AND ON THE STORAGE AND RELEASE OF CATECHOLAMINES IN ISOLATED CHROMAFFIN GRANULES
Authors: Missala K. , Fitt E. , D'Iorio A.
Published: December 2009

The metabolism of adrenaline-β-14C was studied in rats treated with cocaine. The cocaine was administered either as a single subcutaneous injection of 50 mg/kg or by subcutaneous injection of 50 mg/kg of cocaine on 12 consecutive days.In both cases the pattern of urinary metabolites is similar to the pattern observed for controls.Incubation of rat or ox adrenal medullary granules in the presence of 10−3and 10−2 M cocaine brings about a release of catecholamines without producing a release of adenosine triphosphate or of proteins. A lower concentration of cocaine, 10−4 M, is without effect on the granules. This dose of cocaine is also without effect on the uptake of adrenaline-β-14C by ox chromaffin granules.
REGULATION OF ACETOHYDROXY ACID SYNTHETASE IN STREPTOMYCIN-DEPENDENT ESCHERICHIA COLI K-12
Authors: Polglase W. J.
Published: December 2009

The formation of acetohydroxy acid (AHA) synthetase in streptomycin-dependent Escherichia coli K-12 as a function of antibiotic (dihydrostreptomycin) concentration can be expressed by a Michaelis constant (KM = 2 × 10−4M). Valine exerts end-product repression (partial) of AHA synthetase in competition with the positive regulatory effect of dihydrostreptomycin. In dependent E. coli K-12, growth (as measured by optical density increase) correlated closely with the formation of AHA synthetase. The observed positive effect of dihydrostreptomycin on AHA synthetase formation suggests that derepression of this enzyme may be obligatory in antibiotic-dependent cells.
PROTEOLYTIC ENZYMES IN NORMAL AND DYSTROPHIC MOUSE MUSCLE
Authors: Berlinguet L. , Srivastava U.
Published: December 2009

Proteolytic enzymes extracted from normal and dystrophic mouse muscle were studied, and optimum conditions for their activities were established. It was found that these enzymes were active at two pH values, 7.5 and 9. In normal and dystrophic mice, the enzymatic activity increased with age. When the activities of dystrophic muscle enzymes were compared with those of normal muscle enzymes, the increase was most significant in animals 60–90 days of age. The results obtained when the enzymes extracted from normal or dystrophic muscle were incubated with substrates from normal or dystrophic muscle indicate that the defect in the muscle is due to an increase in the activities of the proteolytic enzymes rather than to a chemical change in the muscle proteins.
MITOCHONDRIAL SWELLING AT ACID pH
Authors: Cereijq-Santaló Roberto
Published: December 2009

The effect of variations of pH on swelling of rat liver mitochondria was studied.It was found that mitochondria swell at acid pH (5.0–6.0) when they are incubated in isotonic solution of potassium chloride. The extent of swelling increased with decreasing tonicity of the medium. However, mitochondria did not swell at acid pH when they were incubated in isotonic sucrose. The inhibitory effect of sucrose was partially released by the addition of electrolytes to the medium.When unbuffered suspensions of mitochondria with different pH values were prepared by the addition of hydrochloric acid or sodium hydroxide to the incubation medium, it was observed that the extent of swelling increased both with acidity (6.0 to 5.0) and with alkalinity (7.0 to 8.0).Titration of the unbuffered mitochondrial suspensions showed a maximum buffering capacity in the ranges of pH values where maximum rate of swelling occurred.These results suggest that the mitochondrion behaves as a combination of osmometer and protein gel.
SYNTHESIS AND CHARACTERIZATION OF 1- AND 2-MONOGLYCERYL ETHERS OF ANTEISO FATTY ALCOHOLS, AND REINVESTIGATION OF BENZYLIDENE GLYCEROL SYNTHESIS
Authors: Serdarevich B. , Carroll K. K.
Published: December 2009

The branched-chain fatty alcohols D-(+)-12-methyltetradecanol (C15-anteiso alcohol) and D-(+)-14-methylhexadecanol (C17-anteiso alcohol) were prepared by reduction of the methyl esters of the corresponding C15- and C17-anteiso fatty acids isolated from the lipids of Listeria monocytogenes. The 1- and 2-monoglyceryl ethers were prepared from these alcohols, and tested for effects on circulating leukocyte levels. The synthesis and isolation of the intermediate benzylidene glycerols were also reinvestigated. Reaction intermediates and products were purified and characterized by column, thin-layer, and gas–liquid chromatography and by nuclear magnetic resonance spectroscopy.
METABOLISM OFN-ACETYL-L-ASPARTIC ACID IN MICE
Authors: Berlinguet L. , Laliberté M.
Published: December 2009

Labeled N-acetyl-L-aspartic acid was injected intraperitoneally and intracerebrally into mice. Its metabolism was studied at different time intervals after injection by collecting CO2and by identifying the radioactive metabolites in urine. The localization of injected labeled N-acetyl-L-aspartic acid and of L-aspartic acid was studied by autoradiography. The results show that N-acetyl-L-aspartic acid does not cross the blood–brain barrier, is rapidly metabolized, and that some is excreted unchanged through the kidneys. It can be concluded that N-acetyl-L-aspartic acid is synthesized in the brain and is rapidly metabolized in the body.
AROMATIC METABOLISM IN PLANTS: II. ENZYMES OF THE SHIKIMATE PATHWAY IN SUSPENSION CULTURES OF PLANT CELLS
Authors: Gamborg Oluf L.
Published: December 2009

Liquid suspension cultures were established from tissues of soybean root, wax-bean root, rose stem, reseda stem, horseradish petiole, potato tuber, buckwheat hypocotyl, and of cotyledon, root, and hypocotyl from mung bean. The rose and the reseda cultures required no external supply of growth regulators. Methods of culture and preparation of cell-free extracts are described. Extracts from the cells were assayed for isocitrate dehydrogenase (EC 1.1.1.42), malate dehydrogenase (EC 1.1.1.37), quinate dehydrogenase (EC 1.1.1.24), dehydroquinate dehydratase (EC 4.2.1.10), shikimate dehydrogenase (EC 1.1.1.25), prephenate dehydrogenase (EC 1.1.1.), phenylalanine transaminase (EC 2.6.1.), and phenylalanine ammonia-lyase (EC 4.3.1.5). All of the enzymes were detected in the cultures from mung bean. The isocitrate and shikimate dehydrogenases, dehydroquinate dehydratase, and phenylalanine ammonia-lyase were present in all the cultures. The prephenate dehydrogenase was detected in the mung-bean cultures and the root cultures of soybean and wax bean. Quinate deltydrogenase was detected only in the mung-bean cultures but this represents the first time this enzyme has been found in a plant.
TERATOGENIC COMPOUNDS OF VERATRUM CALIFORNICUM (DURAND): I. PREPARATION AND CHARACTERIZATION OF FRACTIONS AND ALKALOIDS FOR BIOLOGIC TESTING
Authors: Keeler Richard F. , Binns Wayne
Published: December 2009

The isolation and identification of four known alkaloids (jervine, veratrosine, pseudojervine, and isorubijervine) was achieved from teratogenic fractions of Veratrum californicum. Two additional alkaloids, not previously reported and herein designated alkaloids X and V, were also isolated from these fractions and partially characterized.
PHOSPHONOLIPIDS: VI. CHEMICAL AND ENZYMATIC DEGRADATION FOR STUDY OF STRUCTURE
Authors: Baer Erich , Stanacev Nikola Z.
Published: December 2009

The degradation of phosphonic acid analogues of L-α-lecithins and L-α-ceohalins by acetolysis and by lecithinase C from Clostridium welchii was investigated. The acetolysis of both types of phosphonolipids gives in good yields aceto-α,β-diglycerides. Lecithinase C hydrolyzes the phosphonic acid analogues of L-α-lecithins with the formation of D-α,β-diglycerides, but under the same experimental conditions does not react with the phosphonic acid analogues of L-α-cephalins, Thus L-α-lecithins and L-α-cephalins, and the corresponding phosphonic acid analogues behave in an identical manner towards these reagents.
PHYSICOCHEMICAL PROPERTIES OF S-ZONE AND BASE PROTEINS IN GLUTEN
Authors: Oh Yang H. , Sanders Benjamin E. , Gehrke Charles W.
Published: December 2009

Some of the physicochemical properties (solubility, viscosity, sedimentation) and the amino acid composition of the isolated S-zone and base proteins from soft-wheat gluten were studied. The S-zone proteins are homogeneous in size, of low molecular weight (about 20,000), and high intrinsic viscosity in buffer of pH 3,5, having stable structures whose unique solubility properties are related to their atypical amino acid content. The base protein molecules exist in a more folded conformation, having the properties of globular protein molecules. The protein–protein interactions, aggregation, and unfolding conformation of S-zone proteins are major contributing factors at neutral pH to the rheological properties of wheat flour, such as viscosity, elasticity, and cohesiveness.
A QUANTITATIVE METHOD FOR THE DETERMINATION OF CHOLINESTERASE ACTIVITY AFTER STARCH-GEL ELECTROPHORESIS
Authors: Funnell H. S. , Oliver W. T.
Published: December 2009

The method described in this paper is based upon hydrolysis of the starch gel by sodium hydroxide, which releases the dye. The dye is then taken up in an immiscible solvent, and the color determined spectrophotometrically. The method gave good reproducibility in studies both of natural activity and competitive inhibition without requiring any changes in the usual methods. The amount of dye in the final solution was related to the activity towards the dye-producing substrate of the enzyme present in the gel section.
ESTROGEN SYNTHESIS IN NORMAL AND HYPOGONADAL MEN: QUANTITATIVE STUDIES ON THE PRECURSOR ROLE OF TESTOSTERONE
Authors: Epstein B. J. , Raheja M. C. , Frow E. , Morse W. I.
Published: December 2009

Tritiated (7α-3H) testosterone was injected intravenously into normal and hypogonadal men after which all urine was collected for 5 days. The urine was assayed for estrone, 17β-estradiol, estriol, and 3H associated with these fractions by the new method of Blair and co-workers. The specific activities of the urinary estrogen fractions from the normal men were sufficiently low to warrant the conclusion that there were other sources of estrogen production in addition to the plasma testosterone pool. It was also shown that the higher specific activities observed in this laboratory during a previous study, similar except that the Bauld method was used for the assays, were due to radiochemical impurities in the final estrogen fractions. The hypogonadal men had low urinary estrogen levels but there was no evidence of gross abnormality in the metabolism of 3H-testosterone to 3H-estrogens in this group.In two out of three instances, 1,2-3H-testosterone injection produced less 3H-labeled urinary estrogen than 7-3H-testosterone injection. This finding suggested that 3H was lost either from the C-1 or the C-2 position during the biosynthesis of estradiol from testosterone.
PASSIVE CUTANEOUS ANAPHYLAXIS INDUCED IN GUINEA PIGS BY INSULINS AND THEIR COMPONENT CHAINS
Authors: Wilson S. , Aprile Marie A. , Sasaki Louise
Published: December 2009

Small amounts of purified ox insulin and its component chains provoked passive cutaneous anaphylaxis (PCA) in guinea pigs previously injected intradermally with antiserum to ox insulin. Cod insulin and its chains also yielded a PCA reaction with antiserum to cod insulin, indicating that antibody-combining sites are located on both the A- and B-chains of insulin Enzymic digestion of the chain preparations essentially destroyed their ability to combine with anaphylactic antibodies, although the B-chain digests had some residual PCA reactivity.Ox and cod insulins gave positive PCA reactions with the heterologous antiserum but at higher levels than with the homologous antiserum, and none of the four chain preparations reacted with the heterologous antiserum. These results demonstrated differences between the antibody-combining sites on the chains of ox and cod insulins.Insulins from a wide range of species also yielded a PCA reaction with antisera to cod and ox insulins, suggesting that the different hormone preparations have certain antigenic determinants in common. Immunologic relationships between several insulins were derived from the competitive inhibition of one antiserum by the other when both were present in the same animal.
ISOTOPE EFFECTS IN METABOLISM OF14N AND15N FROM UNLABELED DIETARY PROTEINS
Authors: Gaebler O. H. , Vitti Trieste G. , Vukmirovich Robert
Published: December 2009

The purpose of this study was to determine whether the excess of15N present in protein nitrogen of rat tissues is greater than that in nitrogen of the proteins consumed by the animals.Two groups of rats were fed diets in which the protein components were casein and soybean protein respectively. Both the natural excess of15N and its distribution are different in these two proteins. In both groups of rats, abundance of15N in nitrogen of liver and muscle protein exceeded that in dietary protein nitrogen after 2 weeks. When the same diets were continued for 8, 26, or 50 weeks, further accumulation of15N in tissue proteins was slight and limited to a few amino acids. Protein nitrogen of nuclear, mitochondrial, microsomal, and soluble subcellular liver fractions contained essentially the same excess of15N.Analyses of nitrogen from eight nonessential amino acids (proline, glutamic acid, alanine, aspartic acid, argmine, glycine, serine, tyrosine) and six essential ones (valine, leucine, lysine, histidine, phenylalanine, threonine) isolated from dietary and tissue proteins showed that15N accumulated primarily in the nonessential group, and in leucine and valine.15N excess appeared to be greatest in amino acids which are extensively involved in nitrogen transfer. In threonine isolated from liver protein, abundance of15N was below that of dietary threonine, and this deficit increased with duration of feeding in both groups of rats.
KINETIC STUDIES ON THE MECHANISM OF CYTOPLASMIC L-α-GLYCEROPHOSPHATE DEHYDROGENASE OF RABBIT SKELETAL MUSCLE
Authors: Black William J.
Published: December 2009

Studies on initial velocity and product inhibition were carried out on crystalline cytoplasmic NAD+-linked L-α-glycerophosphate dehydrogenase from rabbit muscle, at pH 7.8 and 9.0 at 26 °C. Michaelis and inhibition constants for all the reactants were determined. The kinetic data were consistent with an ordered mechanism in which nicotinamide–adenine dinucleotide (NAD+) or its reduced form (NADH) is bound to the enzyme before the addition of the glycerophosphate (LαGP) or dihydroxyacetone phosphate (DHAP) respectively. At high concentrations NADH, DHAP, and LαGP, but not NAD+, produced substrate inhibition. Combined product-inhibition and dead-end inhibition studies indicated the formation of inactive dead-end complexes of NADH–enzyme, DHAP–enzyme, and LαGP–enzyme–NADH. The low rate constant calculated for the dissociation of the active NADH–enzyme complex suggested an ordered mechanism involving either the formation of an inactive dead-end NADH–enzyme complex or an isomerized NADH–enzyme complex. A choice between these possibilities could not be made on the basis of the present kinetic data. A mechanism for substrate inhibition involving two NAD+-binding sites per mole of enzyme is proposed. Alterations of the ultraviolet absorption spectrum of the enzyme by NAD+ and NADH were in agreement with the conclusion from the kinetic results that the coenzymes are bound to the enzyme before the substrates. DHAP and LαGP caused no alteration in the enzyme spectrum. Spectral changes compatible with the formation of ternary and dead-end complexes were also detected.
STUDIES ON THE LIVETINS OF EGG YOLK: III. HETEROGENEITY AND IMMUNOELECTROPHORETIC BEHAVIOR OF BETA-LIVETIN
Authors: Hui Shun-Fong , Common R. H.
Published: December 2009

Starch-gel electrophoresis of the total livetins of hen's egg yolk resolved 16 zones: seven major zones, six minor zones, and three faint, diffuse zones. One zone was identified with the major component of paper electrophoretic alpha-livetin and hence with serum albumin. Four of the major zones were identified with the major components of paper electrophoretic beta-livetin on the one hand, and with an electrophoretically heterogeneous livetin antigen (livetin antigen 3) on the other hand, thus establishing the electrophoretic heterogeneity and relative immunological homogeneity of the paper electrophoretic beta-livetin fraction. The other two major starch-gel electrophoretic zones were identified as transferrins by their positive staining reaction for iron and comparison of their mobilities with two corresponding serum starch-gel fractions.
EVALUATION OF PROTEIN IN FOODS: XII. EFFECTS OF CALORIE RESTRICTION
Authors: Rao M. Narayana , Morrison A. B.
Published: December 2009

The effects of calorie restriction on net protein utilization (N.P.U.) were studied with proteins of different nutritional value. When protein intake remained constant, reduction in the intake from 32 to 16 kcal per day resulted in a rapid decline in N.P.U. values. Increases in body weight were directly proportional to calorie intake per kg0.73 body weight. Even under conditions of marked caloric restriction, only 70–75% of the ingested protein was used for energy purposes, the rest evidently being used in essential anabolic activities. Studies on carcass composition showed that animals on restricted calorie intakes tried to adapt through shifts in metabolism, and utilized fat in preference to protein for caloric requirements. RNA metabolism in liver and muscle was influenced by energy and protein intakes.When the percentage of calories from protein remained constant, N.P.U. values were not markedly altered until the calorie intake reached the maintenance energy requirement of 150 kcal per kg0.73 body weight. Below this point, N.P.U. values dropped sharply, the rate of decrease being greater for proteins of higher nutritive value. Differences in the nutritive value of proteins, as shown by their ability to satisfy protein requirements in protein-depleted rats, still existed even at restricted intakes of calories.
BIOLOGICAL ACTIVITY OF CORTICOSTERONE AND 11-DEOXY-CORTICOSTERONE C-21 ESTER SULFATES COMPARED WITH THAT OF THE CORRESPONDING STEROID-FREE ALCOHOLS
Authors: Klein G. P. , Giroud C. J. P.
Published: December 2009

Corticosterone-21-suifate was assayed for liver-glycogen deposition in the adrenalectomized mouse, and 11-deoxycorticosterone-21-sulfate for mineralocorticoid activity (urinary sodium/potassium) in the adrenalectomized rat. Compared with equimolar doses of corticosterone (140–870 mμmoles/mouse) and of 11-deoxycorticosterone (4.5–36.0 mμmoles/rat) the two conjugates were devoid of biological activity.
POSITIONAL DISTRIBUTION OF FATTY ACIDS IN THE FATS OF A POLAR BEAR AND A SEAL
Authors: Brockerhoff H. , Hoyle R. J. , Huang P. C.
Published: December 2009

Fats of a polar bear and a seal were analyzed. It was found in both samples that the polyenoic fatty acids preferably occupy the α-positions and, especially, position 3. Position 2 contains short acids, and position 1 contains the longer and more saturated acids. It is suggested that this distribution pattern is typical for mammals feeding on marine fats.
USE OF LIVER HOMOGENATES FOR STUDIES ON THE INTRACELLULAR SITE OF UBIQUINONE BIOSYNTHESIS
Authors: Phillips W. E. J. , Hidiroglou M. , Hatina G.
Published: December 2009

Mevalonic acid was incorporated into the ubiquinone and sterol fractions of rat-liver homogenates. The incorporation of mevalonate was increased approximately 10-fold in the total non-saponifiable matter including sterol by use of the in vitro homogenate system as compared with intact rats. The incorporation into the ubiquinones was decreased. The addition of potential ring precursors, p-hydroxybenzoic acid, L-phenylalanine, or L-tyrosine to the homogenate system did not increase incorporation of mevalonate into ubiquinone.Incubation of subcellular fractions from rat-liver homogenates demonstrated that supernate plus either mitochondria or microsomes were necessary for the biosynthesis of ubiquinone.
THE SYNTHESIS AND PROPERTIES OF ADENOSINE-2′ DIPHOSPHATE AND ADENOSINE-3′ DIPHOSPHATE
Authors: Mitchel R. E. J. , Ward D. C. , Tener G. M.
Published: December 2009

Adenosine-2′ diphosphate (A2'DP) and adenosine-3′ diphosphate (A3′DP) have been prepared by reaction between the corresponding mononucleotides with phosphoramidic acid in formamide. The products were isolated by ion-exchange chromatography on DEAE-cellulose and characterized by enzymic and chemical methods. A major side reaction in the synthesis is the phosphorylation of unprotected hydroxyl groups oil the nucleotides. A2′DP and A3′DP were found to be more labile than adenosine-5′ diphosphate, and to decompose at neutral pH by direct hydrolysis of the pyrophosphate bond as well as by formation of adenosine-2′,3′ cyclic phosphate. They are stable in 10% trichloroacetic acid at 0° for 10 minutes. Phosphomonoesterase degrades both to nucleosides by way of the corresponding nucleoside monophosphates. Ribonuclease T2does not attack A2′DP but degrades A3′DP to adenosine-3′ phosphate. Neither nucleotide is degraded by snake venom or spleen phosphodiesterases under conditions where oligonucleotides are completely hydrolyzed.
CATIONIC ANTIBIOTICS AND PHOSPHOLIPASE C AS TOOLS IN THE STUDY OF PHOSPHOLIPID STRUCTURE AND FUNCTION: II. COMBINED EFFECTS OF COLISTIN SULFATE AND PHOSPHOLIPASE C ON THE IN VITRO BLOOD CLOTTING SYSTEM
Authors: Ottolenghi Abramo C.
Published: December 2009

The phospholipases C from Bacillus cereus and Clostridium perfringens showed different affinities for the thromboplastic suspensions used in the one-stage prothrombin time system. In the presence of colistin sulfate the affinities were reversed. In each case one enzyme would further degrade the thromboplastic material following destruction by the other enzyme. The results indicate that a negative zeta potential of the micelles is probably optimal for both clotting activity and susceptibility to B. cereus enzyme, whereas the opposite charge inhibits clotting but renders the substrate more susceptible to C. perfringens enzyme.
FACTORS INFLUENCING TETRAZOLE REDUCTION BY REDUCED NICOTINAMIDE–ADENINE DINUCLEOTIDE IN PIGEON-HEART MITOCHONDRIA
Authors: Talesara C. L. , Blanchaer M. C.
Published: December 2009

The effect of adenosine triphosphate, adenosine diphosphate, adenosine monophosphate and inorganic phosphate on the reduction of 2-(p-iodophenyi)-3-p-nitrophenyl-5-phenyl tetrazolium chloride (INT) to its formazan by reduced nicotinamide-adenine dinucleotide (NADH) was studied in pigeon-heart mitochondria. Formazan production was followed at 540 mμ in 2.2 ml medium containing 0.4–0.5 mg mitochondrial protein, 0.22 M mannitol, 0.067 M sucrose, 0.02 M Tris–chloride, 0.02 mM EDTA, 0.5–3.0 mM INT, and 38 μM NADH at pH 7.2 and 28 °C. By means of the respiratory inhibitors Amytal, rotenone, antimycin A, and cyanide, it was shown that INT diverts electrons from the respiratory chain principally at the flavoprotein level. In contrast to its inhibitory effect on "the O2-linked oxidation of NADH, 10 mM adenosine triphosphate stimulated the reaction rate and formazan yield in the present system. Equimolar inorganic phosphate also increased the initial velocity but adenosine diphosphate and adenosine monophosphate did not. Preliminary kinetic studies suggest that NADH, but not INT, combines with the form of NADH dehydrogenase in the respiratory chain with which adenosine triphosphate reacts.
BIOSYNTHESIS OF PORPHYRINS IN WHEAT LEAVES: II. 5-AMINOLAEVULINATE HYDRO-LYASE
Authors: Nandi D. L. , Waygood E. R.
Published: December 2009

5-Aminolaevulinate hydro-lyase (EC 4.2.1.24), which catalyzes the formation of porphobilinogen from 5-aminolaevulinate (5-ALA), was isolated from wheat leaves and partially purified. The enzyme was specific for 5-ALA, sulfhydryl-dependent, and required divalent cations for maximum activation. Pyrophosphate, EDTA, and ATP were strongly inhibitory. Laevulinate, but not ethyl laevulinate, was also an inhibitor. The pH optima were 7.5–7.6 in Tris and 7.2–7.8 in phosphate buffer. Michaelis constants for 5-ALA, Mg++, and Mn++were 1 × 10−3 M, 3.1 × 10−4 M, and 3.7 × 10−5 M respectively. The enzyme was localized partially in the chloroplasts and also in 'remaining particles'. Inhibition studies indicated that a carbonyl group γ to an ionized carboxyl group is necessary for binding the substrate to the active site.
FISH-MUSCLE PURINE AND PYRIMIDINE NUCLEOSIDE PHOSPHORYLASES
Authors: Tarr H. L. A. , Roy Joan E.
Published: December 2009

Three purine nucleoside phosphorylase preparations (isoenzymes) were obtained by ammonium sulfate fractionation and DEAE-cellulose chromatography of aqueous extracts of lingcod muscle. Dialysis, adsorption on alumina Cγ, and elution with 0.4 M phosphate buffer yielded further purification. The most active enzyme preparation had about 120 times the activity of initial extracts. It utilized hypoxanthine, 6-mercaptopurine, guanine, 8-azaguanine, xanthine, adenine, 2,6-diaminopurine and 6-methylpurine in presence of ribose 1-phosphate or deoxyribose 1-phosphate. Several substituted purines were not utilized and did not inhibit the reaction between hypoxanthine and the pentose phosphates. The Kmwith inosine as substrate was 3.2 × 10−6 M. A pyrimidine nucleoside phosphorylase, distinct from the purine nucleoside phosphorylase, occurred in the DEAE-cellulose fraction comprising one of the purine nucleoside phosphorylases. Its activity was much lower than that of the purine nucleoside phosphorylase preparations. Uridine and thymidine were the best substrates. Deoxyuridine was a poor substrate, and neither cytidine nor deoxycytidine was utilized. The equilibrium with all preparations was about 80% in favor of nucleoside formation. The purified enzymes were all destroyed by freezing.
FATTY ACIDS IN DROSOPHILA MELANOGASTER: III. CHARACTERIZATION OF COMPONENT FATTY ACIDS BY PAPER CHROMATOGRAPHY
Authors: Kato Mikio
Published: December 2009

It has been previously reported that the interference of unsaturated fatty acids with the chromatographic identification of saturated fatty acids may be minimized by mercuration. Because simple hydrazones of unsaturated fatty acids and their mercurated derivatives give different color reactions on treatment with diphenylcarbazone, the component saturated and unsaturated fatty acids in pupal lipids of Drosophila melanogaster were identified on the same chromatographic paper. The amounts of individual fatty acids were calculated from the spectrophotometric data by an easier method than that described in the previous reports. The quantities of saturated and unsaturated acids in different strains of D. melanogaster appeared to be related to their genetic nature.
THE INCORPORATION OF14C-THREONINE AND14C-GLUCOSAMINE INTO SUBCELLULAR FRACTIONS AND INTO BOVINE SUBMAXILLARY MUCIN BY SLICES OF BOVINE SUBMAXILLARY GLAND
Authors: Lawford G. Ross , Schachter Harry
Published: December 2009

Bovine submaxillary gland slices were used to investigate the subcellular site of biosynthesis of bovine submaxillary mucin (BSM). Slices were incubated with14C-threonine and14C-glucosamine in Krebs medium III at pH 7.4 and 37 °C for varying periods of time up to 180 minutes. Methods of subcellular fractionation based on those developed for liver proved unsatisfactory for use with submaxillary gland, probably because of the presence of large amounts of highly viscous mucin. A method of subcellular fractionation was developed which yielded five fractions: P (total soluble protein) and C (soluble BSM, purified by Cetavlon precipitation) were prepared from the 16,000 × g supernatant; and R (ribosomes), S (deoxycholate-soluble membrane-bound protein), and a residual fraction D were prepared from the 16,000 × g pellet. The incorporation of both tracers into these fractions was time-dependent and was inhibited by preincubation of the slices for 15 minutes with puromycin. The highest specific activities were recorded in the R and S fractions, implicating them as possible sites of glycoprotein biosynthesis. The time course of14C-threonine incorporation suggested that protein is made on ribosomes, is transferred to the endoplasmic reticulum, and is then ready for storage or secretion. The time course of14C-glucosamine incorporation suggested that carbohydrate is linked to the protein both while the protein is still associated with the ribosomes and after it has been released from them. The involvement of ribosomes in carbohydrate attachment to polypeptide received additional support from zone centrifugation analysis on sucrose-density gradients of ribosomes labeled with14C-glucosamine.
1-AMINOCYCLOPENTANECARBOXYLIC ACID, AN INHIBITORY ANALOGUE OF METHIONINE IN ESCHERICHIA COLI
Authors: Abshire C. J. , Larouquere J. , Berlinguet L.
Published: December 2009

1-Aminocyclopentanecarboxylic acid (ACPC) inhibits the growth of Escherichia coli 9723 at a concentration of 5 mg/5 ml assay. This inhibition may be reversed by L-methionine and DL-methionine but not by D-methionine. The data show that the inhibition is competitive, the inhibition ratio for ACPC:L-methionine being 20:1. Studies with labeled ACPC and methionine seem to verify that the toxicity of ACPC is due to an intracellular mechanism.
CHOLINESTERASE AND MONOAMINE OXIDASE ACTIVITIES IN SKELETAL MUSCLE OF NORMAL AND HEREDITARY DYSTROPHIC MICE
Authors: Srivastava Uma , Berlinguet Louis
Published: December 2009

Acetyl- and butyryl-cholinesterase activities in dystrophic mouse muscle are increased significantly, whether the results are expressed in relation to (a) the fresh weight or (b) the total nitrogen, of the muscle. Activities of these enzymes do not show any change in normal and dystrophic mouse liver and brain. However, in dystrophic lens, the enzymatic activity is decreased when the results are expressed on the basis of fresh weight. This is because of a change in the chemical composition of the lens since no significant change in both enzymatic activities could be observed when the results were expressed on the basis of lens protein. This increase in the cholinesterase activity of dystrophic muscle is not incompatible with the change in the distribution pattern of these enzymes in the muscle noted by many workers, or to the high sensitivity of dystrophic muscle to acetylcholine.Monoamine oxidase activity in dystrophic mouse muscle showed a significant increase when the results are expressed in relation to (a) the fresh weight or (b) the total nitrogen, of the muscle. This increase in activity could be due to enzyme induction, i.e. increase in the amine content followed by an increase in monoamine oxidase activity in dystrophic muscle.
THE MACROMOLECULAR COMPOSITION OF HEN'S EGG YOLK AT SUCCESSIVE STAGES OF MATURATION
Authors: Mackenzie S. L. , Martin W. G.
Published: December 2009

Yolk at different stages of its formation was separated into its major macromolecular fractions, namely, granules, low-density fraction (LDF), and water-soluble fraction (WSF), and the components of these fractions were examined by ultracentrifugal, electrophoretic, and chromatographic methods. Total yolk solids and the amounts of the several fractions all increased exponentially during yolk formation but at differential rates for different fractions. The proportion of LDF increased rapidly to a maximum (76% of total solids) for yolks weighing 2–5 g, and then declined. Both the WSF and granule fractions decreased in proportion during the early period of yolk formation and then increased slightly as yolk neared maturity. Of the WSF components, the proportions of α-livetin (serum albumin) and β-livetin increased somewhat and γ-livetin (serum γ-globulin) decreased slightly, during yolk maturation. The major granule subfraction, PvLv, showed an increased proportion of phosvitin and α-lipovitellin relative to β-lipovitellin during yolk formation. This increase in the proportions of the phosphorus-rich proteins was not paralleled by a corresponding increase in PvLv phosphorus, which suggests that other phenomena may be involved.