Animal Tissue Extracts Research Articles

A Precipitation Reaction Between Human Serum and a Soluble Tissue Component

Summary Certain extracts of human and animal tissues have been found to give a precipitation line with normal human serum in a gel diffusion system. This precipitation reaction results from the interaction of a basic tissue component, probably a protein, with human serum albumin. Evidence has been presented that the reaction is not of the immune type and that the interaction, which shows specificity for human albumin, is due primarily to electrostatic forces.

Occurrence and quantitative determination of 2-dimethylaminoethanol in animal tissue extracts.

RECENTLY the function of 2-dimethylaminoethanol (Deanol) in the biosynthesis of choline and its role as a possible precursor of cerebral acetylcholine1 has intensified interest in this substance. Levine and Chargaff2 could not find it in phospholipides. Artom and Crowder3 claimed the occasional occurrence of traces of it in rat liver. Recently Wolf and Nyc4 reported that they were able to isolate it from a mutant strain (47904) of Neurospora crassa, which accumulates this substance. In a preliminary paper, Venkataraman and Greenberg5 showed on the basis of chromatographic evidence, that incubation of a rat liver extract with aminoethanol and formaldehyde yielded methylated intermediates of choline. To clarify the question of the natural occurrence of 2-dimethylaminoethanol in animal tissue a method was developed for the isolation, characterization and quantitative determination of small amounts of this substance in the presence of other amines.

Determination of Vitamin B12 With an Escherichia Coli Mutant

Abstract An improved method for the determination of vitamin B12, using a more sensitive mutant of Escherichia coli, which enables determination of 4-20 µµg./ml. of the vitamin, has been described. Comparative determinations of vitamin B12 content of human sera and of animal tissue extracts gave similar values when assayed with both E. coli and O. malhamensis. Although O. malhamensis is more specific in its response to cyanocobalamin than E. coli, the latter organism seems just as reliable for determination of vitamin B12 in serum and in tissue of human and animal origin. The simplicity, rapidity, and sensitivity of the improved E. coli assay make it very suitable for clinical use.

Transaminations with L-glutamate and alpha-oxoglutarate in fresh extracts of animal tissues.

Transaminations with L-glutamate and alpha-oxoglutarate in fresh extracts of animal tissues.

Studies on pigment-activating substances in animals I. The separation by paper electrophoresis of chromactivating substances in arthropods

It has been established that many substances can bring about a change in position of pigments in crustacean chromatophores (Florey, 1951). The extracts of certain animal tissues (pituitary, crustacean sinus-gland and post-commissure organs, insect brain and corpora cardiaca) are especially active (Brown, 1940; Knowles, 1953; Dupont-Raabe, 1952; Thomsen, 1943); there is evidence that these tissues intervene in the normal colour change of the animals which possess them and that their products may properly be considered as hormones. On the other hand, many species which do not themselves possess chromatophores (oligochaetes, molluscs, and many insects) have nevertheless been shown to contain substances in their tissues which will, after injection into crustaceans, initiate pigment movements (Scharrer, 1954). It has not yet been ascertained whether these pigment-activating substances chemically resemble normal colour-change hormones or whether the pigment movements they produce are pharmacodynamic effects irrelevant in the study of colour physiology.

Studies on Pneumococcal Polysaccharide

Summary Biosynthesis of C14-SI is easily accomplished by the addition of C14-glucose to the culture medium in which the pneumococci are grown. Approximately 4.5 per cent of the radioactivity thus introduced into the medium is present in the purified C14-SI. Introduction of 100 μc of C14-glucose per liter of culture medium is sufficient to adequately label SI for identification in animal tissue extracts when the dosage is 500 μg per mouse. If smaller antigenic dosages are to be used additional C14-glucose would probably be required for adequate labeling if the counting technique described is to be used. Significant comparative counts due to C14-SI can be secured on splenic tissue extracts by making the determinations on material in water solution if the dosage approximates 500 μg per mouse. This method permits use of extracts for antigenic assay or other biological procedures not possible when the sample is destroyed as in the barium carbonate method of determining total radioactivity. Results obtained by this counting technic are comparative only but are proportional to the concentration of C14-SI present in the sample.

Enzymatic Hydrolysis of the —S—CO— Linkage

During biological and biochemical studies of a new derivative of thiamine, O,S-diacetyl thiamine, synthesized by Matsukawa and Kawasaki1, it was found that this substance was readily hydrolysed to thiamine and acetic acid by liver extracts2. O,S-Di-acetyl thiamine: was found in the earlier work to be almost as efficacious as thiamine by the prevention assay of B1-deficiency in Uroloncha domestica. Although a number of reports have appeared concerning the hydrolysis of ordinary esters by various esterases, no investigation seems to have been carried out on the enzymatic hydrolysis of the —S—CO— linkage except that on the “succinyl and acetyl coenzyme A deacylases” reported by Gergeley et al. 3. We have therefore studied the hydrolysis of lower thioesters, such as methyl thioacetate and ethyl thioacetate, by animal tissue extracts, comparing them with methyl acetate and ethyl acetate. Manometric determination was used for enzyme assays in much of this work.

The pathways of acetate oxidation

In micro-organisms considered to oxidize acetate via the dicarboxylic acid cycle or by oxidation to glycolic acid, there were found the enzymes for the synthesis of citric and for the oxidation of isocitric acid. The culture media of moulds believed to oxidize acetate via glycolic acid contained citric, α-ketoglutaric, succinic, malic, and glycolic acids. In most of these organisms malonate inhibited acetate oxidation. In the absence of malonate, citric acid was detected chromatographically from solutions where acetate was oxidized whereas in the presence of malonate only succinate was detected. Formation of succinate from acetate seems to occur by the oxidative condensation of acetylcoenzyme A to succinate. Cell-free extracts of bacteria, moulds, yeast, and animal tissues contain the enzyme which oxidizes acetate in the presence of coenzyme A, flavine adenine dinucleotide, DPN, and Mg +2. It is postulated that all cells oxidize acetate via citric acid cycle but also possess the dicarboxylic acid cycle which provides the oxalacetate required for citric acid formation.

Conversion of alcoholic to lactic fermentation in yeast extracts by animal tissues

1. 1. The addition of brain tissue extracts to a yeast extract capable of rapid alcoholic fermentation at pH 6.0 brings about a fall in the rate of carbon dioxide evolution in the presence of sucrose. 2. 2. The inhibition of yeast fermentation is due to a thermolabile constituent of the brain tissue extract. There is no evidence that the inhibition is due to destruction of a yeast dehydrogenase. 3. 3. Lactic acid accumulates when a brain tissue extract is incubated with an extract of yeast. 4. 4. The inhibitory action of a brain tissue extract on yeast fermentation of glucose is partially neutralized by the presence of nicotinamide (0.011 M). The rate of formation of lactic acid is also greatly increased under these circumstances. 5. 5. Lysed red blood cells, extracts of tumors, and dialyzed muscle extracts have similar effects to that of a brain extract in converting alcoholic fermentation of yeast (extracts) into glycolysis. 6. 6. The results are explained by the presence in animal tissue extracts of diphosphopyridine nucleosidase (DPN-ase) and diphosphopyridine nucleotide (DPN)-linked lactic dehydrogenase. The former enzyme causes a drop in alcoholic fermentation due to destruction of DPN. The latter enzyme causes both a drop in the rate of alcoholic fermentation and a rise in the rate of lactic acid formation owing to its competition with carboxylase for pyruvic acid produced during the breakdown of glucose. It is the circumstance that the lactic dehydrogenase of animal tissues is DPN-linked, whereas that of yeast is not DPN-linked, which is largely responsible for the fact that in a mixture of yeast and animal tissue extracts the process of alcohol formation from glucose is largely converted to glycolysis.

Displacement chromatography on ion-exchange columns of the carboxylic acids in plant tissue extracts.

THE importance of many carboxylic acids in metabolism makes it desirable that techniques for their estimation should, in general, be applicable to extracts of plant and animal tissues. It is essential that such extracts containing low concentrations of the significant acids should be as free as possible from inorganic salts, sugars and pigments. Such impurities, because of their relative abundance, can seriously detract from the value of paper chromatograms obtained using the technique of Lugg and Overell1, which otherwise provides a rapid and convenient method of simultaneous separation and identification of carboxylic acids. Even the customary ether extraction of plant tissues does not always satisfy these purity requirements.

Distribution of the Protective Factors for Vitamin C in Fractions of Liver Homogenates of the Rat

IT has been reported by de Caro and Giani1, Mawson2 and Giri and Shouri3 that animal tissue extracts contain factors which protect vitamin C against oxidation. These protective factors were found to be of the nature of thiols1. Mawson observed that liver extracts from which glutathione was removed by prolonged dialysis retained the protective properties of fresh extract and concluded that glutathione is not the only constituent responsible for the protective mechanism existing in animal tissues. Later, Giri and co-workers4 reported that certain purines such as xanthine, uric acid, adenine, guanin, theophylline and nucleic acids exert a protective action on vitamin C against oxidation. Since purines and nucleic acids are widely distributed in cells, it seemed of interest to find out the distribution of the protective factors for vitamin C in various fractions of liver homogenates, in the hope of gaining an insight into the nature of the factors responsible for the protective action.

Some Experiments on the Microscopical Demonstration of Zymohexase in Animal Tissues

ABSTRACT The presence in extracts of animal tissues and yeast of an enzyme catalysing the cleavage of hexosediphosphate (fructofuranose-1:6-diphosphate) into two molecules of triosephosphate was demonstrated by Meyerhof & Lohmann (1934a):

THE EFFECT OF COLAMINE ON GROWTH AND PROTOPLASMIC STREAMING IN AVEN

AS FAR as is known, there is no substance in animals comparable to the auxins which control the growth of plants. Extracts of animal tissues, however, contain substances which inhibit the growth of animal cells in tissue culture. Among these inhibiting substances is colamine, or ethanol-2-amine. The history of the study of inhibitors extracted from liver is surveyed by Medawar (1937). In brief, liver extracts have been known for some time to have an inhibitory effect on cells in tissue culture. In 1926, Heaton made the first exhaustive studies of this factor in aqueous or dilute alcoholic extracts of adult liver. The extracts inhibited the growth of chick heart fibroblasts but had no effect on a number of different epithelial tissues. Heaton also extracted a similar factor from brewer's yeast, but the inhibition was probably due-to malt present in the yeast extracts, since Heaton himself later showed that extracts of malt contained active inhibitors of this kind. Substances inhibiting the growth of mesenchvme but not epithelium have been extracted frorn germinated and ungerminated barley, from the seeds of maize, wheat, and oats, from whole grain flour and even whole grain bread. The inhibitor has also been obtained in large quantities from whole orauges. All the extracts contained yellow pigments in varying amounts, but removal of the pigment did not affect the activity of the inhibitor. Brues and co-workers (1936) have recently studied the effect of these growth inhibiting factors on animal tissue cultures. They used saline or alcoholic extracts of liver to treat different embryonic and sarcomatous tissues. The radius of growth of cultures of fibroblasts was greatly diminished and there was a decrease in the wandering monocytes, although the life span of the treated cultures was not affected and rare mitoses could be found. Cells were smaller in treated than in untreated material. Epithelium was not affected. Brues et al. (1940) later identified colamine in the inhibitory extract. Colamine produced inhibition of growth in concentrations of 0.25 to 2 mg. per cc. A number of other amines were also active. The following study was undertaken to determine whether colamine has any effect on growth in plants comparable to its action on animal tissues, and to study the effect of colamine on protoplasmic streaming in plants, more especially to study its effect in the presence of auxin.

Mechanism of the Enzymatic Phosphorylation of Thiamin

WE have previously demonstrated that thiamin pyrophosphate may be synthesized from the free vitamin by the action of enzymes present in alkaline washed brewer's bottom yeast, provided that hexose-diphosphate and certain factors present in boiled aqueous extracts of animal tissues are present1,3. Synthesis under these conditions was completely inhibited by 0·005 M. sodium iodoacetate, but was relatively unaffected by 0·04 M. sodium fluoride. More recently3, we have demonstrated that cozymase and acetaldehyde may be substituted for the boiled tissue extract with no change in the degree of synthesis or of the iodoacetate inhibition. Phosphoglyceric acid may also be substituted for the boiled tissue extract, but under these conditions iodoacetate fails to affect the synthesis while fluoride causes complete inhibition3. Using specially prepared alkaline washed yeast, we have found that cocarboxylase synthesis with phosphoglyceric acid is considerably diminished. It may be restored, however, by the addition of catalytic quantities of adenylic acid. As a result of these experiments we have proposed the reaction

The use of a blood coagulant extract from the human placenta in the treatment of hemophilia

Summary Tissue protein extracts prepared from the human placenta have been found to have the same type of effect in the control of local hemorrhage as has been reported from the use of animal tissue extracts. However, a striking difference has been observed in the results with this extract and those reported from the use of animal tissue preparations in the treatment of patients with hemophilia. Animal tissue extracts have shortened the coagulation time of the blood of patients with hemophilia for very brief periods, whereas human placental extracts have reduced the clotting time of both venous and capillary blood for periods varying from forty-eight hours to as long as nine days. Despite repeated slight trauma incident to active childhood, two children with hemophilia have been able to lead normal, active lives for periods of several months by the continued administration of this material at carefully regulated intervals. In the relatively small series of patients with hemophilia herein reported, the blood coagulant extract has been well tolerated by oral administration or by intramuscular injection. It must be emphasized that this blood coagulant extract is fatal when injected intravenously in small laboratory animals. Therefore, in using this material in patients with hemophilia, intravenous injection must be avoided as by this route unfortunate, if not fatal, results might ensue.

Studies On Bacterial Nutrition: II. The Distribution of a Growth Stimulating Factor in Animal and Plant Tissues

In a previous study of the requirements for growth of the more exacting bacteria it was found that a growth stimulating factor1 could be removed from veal infusion. This factor, when added to a simple culture medium, rendered the medium suitable for growth of some of the more fastidious organisms. In certain instances small amounts of added substance amounting to only 0.0006 to 0.00006 milligram of total solids per cubic centimeter of medium were sufficient to permit development in a synthetic medium in which the organisms ordinarily refused to grow.2 Additional study of the factor responsible for this effect seemed desirable. However, before attempting chemical purification of the growth substance obtained from veal infusion, it was considered best to take up first the question of its occurrence in various natural sources. There is considerable evidence in bacteriological literature that growth stimulating substances can be found in a number of animal and plant tissues. In many of these reports, however, it is not clear whether the stimulating effect is due simply to a more abundant supply of readily available food or to the presence of accessory factors which may be required by certain types in addition to the ordinary nutritive material. While no attempt will be made to give a comprehensive survey of the literature, since this has been summarized by Peskett,3 certain references which bear upon the present study should be mentioned. The important role of plant or animal tissue extracts in cultivation of Hemophilus influenzae has been emphasized especially by Davis,4 Thjotta and Avery,5 Fildes6 and others. Substances of a supposedly vitamin-like nature which exerted a strong growth-stimulating activity on hemolytic streptococci and yeast were reported by Freedman and Funk7 in

CHOLESTEROL AND LIPOID PHOSPHORUS IN INFANCY AND IN CHILDHOOD

The elaboration of newer methods for the determination of during the past few years has resulted in an increase in the literature on the subject; notwithstanding this added knowledge, the role played by these substances in the general body metabolism is neither clearly nor fully understood. One of the underlying reasons for this failure to evaluate properly the function of the is the insufficient data as to their normal incidence especially during infancy and childhood. Additional data on the normal values during these periods may be of some aid. The collective term lipoids encompasses a heterogeneous mass of substances which are found after extraction of animal or vegetable tissue with ether or alcohol or a mixture of the two. Some of the substances obtained by extraction are fatty; they yield fatty acids on hydrolysis but differ from neutral fats in that they contain nitrogen or nitrogen and phosphorus;

Opotherapie endocrinienne. Les bases physiologiques, les syndromes, la posologie de l'opotherapie par les glandes a secretions internes.

This volume deals with the general principles of organotherapy. It contains one chapter on the chemistry and the processes of production of extracts of animal tissues; one long chapter on the thyroid and thyroid disorders and their treatment; a brief chapter on parathyroids and the thymus; an exhaustive chapter on suprarenal disorders, and shorter chapters on the hypophysis, the testes and ovarian disorders and their treatment. The chapter on the pancreas deals almost exclusively with insulin, and there is a short but sane chapter on the organotherapy of so-called pluriglandular syndromes. The volume as a whole gives evidence that the author is familiar with the important literature in this field, although reference to this literature is meager. The author further shows good judgment in most cases when dealing with the analysis and conclusions in fields in which great uncertainty still exists both as to etiology and therapy. For example, in

A method for the quantitative determination of small amounts of quinin and quinidin with bromin water

We undertook to learn whether quinin could be determined quantitatively in extracts of animal tissues by means of the so-called thalleioquin color reaction, but it soon became apparent that the intensity of the color varies with too many factors to permit of its use for this purpose. It was observed, however, that the color of the bromin water disappeared sharply on the addition of a slight excess of quinin, and the present method depends upon the fact that when quinin is added to bromin they combine in definite proportions with the loss of the color of the bromin in reflected light.Hart1 attempted unsuccessfully to estimate quinin quantitatively by means of the thalleioquin reaction. He found, however, that four atoms of bromin are absorbed by the quinin, two of them loosely. He says, “It follows, then, that quinine may be determined from its bromine absorption for five minutes in the manner stated: 1 cc. N/10 Br. = 0.0081 g. quinine.” This method has not been employed hitherto so far as we know.Nine sol...