bluuebunny/crossref_metadata_2025_split huggingface dataset:
DEHYDROGENASES IN DEVELOPING BONE IN THE RAT
Authors: FULLMER HAROLD M. 
Published: April 2011

Sections of mandibles from seventy-five 5-day old rats were studied histochemically for malic, isocitric, α-glycerophosphate, glutamic, lactic, 6-phosphogluconic, glucose-6-phosphate, d(—)- β-hydroxybutyric and succinic dehydrogenases and TPN- and DPN-diaphorases. All the enzymes studied (except TPN-glutamic dehydrogenase) were found in osteocytes, osteoblasts and osteoclasts. The results suggest that all bone cells possess fully functional citric acid cycles, pentose cycles, and the capacity to metabolize fatty acids and carbohydrates. Osteoclasts generally exhibited greater enzymatic activity than either osteoblasts or osteocytes indicating a strong broad metabolic capacity. The demonstration of isocitric dehydrogenase in osteoclasts tends to discredit Walker's theory of bone resorption (18).
APPLICATION OF THE STEREOSPECIFIC INHIBITOR L-PHENYLALANINE TO THE ENZYMORPHOLOGY OF INTESTINAL ALKALINE PHOSPHATASE
Authors: WATANABE KEIICHI , FISHMAN WILLIAM H. 
Published: April 2011

The intestine-specific inhibitor of alkaline phosphatase, l-phenylalanine, has been employed in the study of alkaline phosphatase in rat intestine. Five substrates were used, β-glycerophosphate, o-carboxyphenylphosphate, α-naphthyl acid phosphate, naphthol AS-BI phosphate and naphthol AS-TR phosphate. The hydrolytic rate and specific l-phenylalanine inhibition was most marked in the case of the first three substrates. When formol-Ca fixed, gum-sucrose treated sections of rat intestine were incubated in these substrates in the presence separately of 0.05 Md- and l-phenylalanine and Gomori's calcium-cobalt method was employed, a striking inhibition by the l-isomer was regularly found. Azo dye methods were applied under different conditions of time, temperature and cation concentration in some instances. The contrast was marked but not dramatic in the case of α-naphthyl acid phosphate, and it was evident but not marked in the case of naphthol AS phosphate substrates. These findings have been discussed. The alkaline phosphatase of intestinal epithelial cells (human) grown in tissue culture exhibited great sensitivity to l-phenylalanine. Rat kidney and leukocyte alkaline phosphatase were relatively insensitive under the same conditions. Morphologically, the l-phenylalanine-sensitive alkaline phosphatase in rat intestine is largely confined to the striated border of the epithelial cells. In animals maintained on a high fat diet, azo dye methods demonstrated the presence of fine granules in the region of the Golgi apparatus and terminal web in the presence of d-phenylalanine. These reactions were absent in companion sections incubated in l-phenylalanine-containing substrate solutions. The possibility of comparing the morphology of a tissue in which both test and control solutions are identical except for the configuration of one component may contribute to a more precise interpretation of the results obtained in the test section.
A HISTOCHEMICAL METHOD FOR DEMONSTRATING PROTEIN-BOUND SULFHYDRYL AND DISULFIDE GROUPS WITH NITRO BLUE TETRAZOLIUM
Authors: DEGUCHI YASUO 
Published: April 2011

Nitro-BT was applied to an alkaline tetrazolium method at low alkaline pH (pH 8.4) for the demonstration of sulfhydryl and disulfide groups. Evidence of specificity was given in test tube experiments and histochemical techniques. Histochemical findings with the method employed were almost similar to those of Barrnett and Seligman's method, except for a little better contrast on sections, and a fine formazan of nitro-BT gave a strict localization of sulfhydryl groups. A comparative distribution of sulfhydryl and disulfide groups was observed in the skin of foot-sole. The detachment and destruction of sections were prevented by the present method which was advantageous in shortening the reaction time to ⅕-1/10 and in keeping a low pH in the solution.
AMINO ACID CONSTITUTION OF CHROMOSOMES
Authors: SHARMA ARUN KUMAR , CHATTERJI ASOKE KUMAR 
Published: April 2011

The present experiment was designed to demonstrate the presence of two protein-bound amino acids, viz. tryptophan and tyrosine, in the different, cellular constituents with particular reference to chromosomes. The materials chosen were the root-tips of Allium cepa and Vicia faba. Studies were made from the smeared tissue preparation. For tryptophan, indole reactions following two methods, viz., that of Gurr (1958) and of Glenner and Lillie (1957) were followed and the latter yielded satisfactory results; while for tyrosine, Bensley and Gersh's modified Millon's reaction (1953) was adopted. The results of the reactions show that tryptophan is present in much less amount than tyrosine in the protein component of the chromosomes. The same is true with cytoplasm as well. The strongly positive tyrosine reaction of nuclear membrane, with distinct strands towards the cell membrane, has been attributed to its role in nucleocytoplasmic transfer and synthesis of cytoplasmic protein. The less positive reaction of nuclear membrane with tryptophan suggests its comparatively little importance in transfer. Only in case of tyrosine, the differential reaction of the euchromatic and heterochromatic parts could he made out, the latter giving negative response. It has been suggested that it might appear only during the time of transfer through the nucleolus. Tryptophan on the other hand shows uniform distribution throughout the chromosome length.
THE ACTIVITY OF URIDINE DIPHOSPHATE GLUCOSE-GLYCOGEN SYNTHETASE IN SOME EMBRYONIC TISSUES
Authors: GRILLO T. ADESANYA IGE , OKUNO GIICHI , PRICE SWETLANA , FOÁ PIERO P. 
Published: April 2011

Uridine diphosphate glucose (UDPG)-glycogen synthetase has been studied using chemical and histochemical methods. Activity was demonstrated in whole egg and in homogenates of whole 3-day old chick embryos, in the heart and skeletal muscle from the 5th day and in the liver from the 7th day of incubation onwards. The activity increases with age until the 16th day at which time a sharp fall was noted in all tissues studied. Enzyme activity was demonstrated also in heart, liver, skeletal muscle and brain of rat embryos from the 14th day onwards. In these tissues the activity increases with age until birth. The appearance of UDPG-glycogen synthetase in the tissues of the chick embryo coincides with the time at which, according to previously published data, glycogen begins to appear. In the liver of the rat embryo, the enzyme appears before glycogen formation starts. This is contrary to the behavior of phosphorylase which appears later, at least in skeletal muscle. These findings are in agreement with the view that, in vivo, glycogen synthesis occurs via the UDPG, rather than the phosphorylase, pathway.
MONOAMINE OXIDASE DISTRIBUTION IN THE RABBIT EYE
Authors: SHANTHAVEERAPPA T. R. , BOURNE G. H. 
Published: April 2011

Monoamine oxidase distribution in the different components of the eye has been described in detail. The degree of enzyme activity varied from one component of the eye to another, the corneal stroma and Descemet's membrane being completely negative. The corneal epithelium, choroid and nonpigmented cells of the iris stroma showed mild positive activity. Strong positive activity was present in the myelin sheaths of optic and oculomotor nerves and also in the ciliary epithelium. The irido-corneal angle (meshwork) showed a moderately strong positive activity against the surrounding tissues. The most interesting observations were made in the retina. The pigment epithelial cell layer and the outer plexiform layer showed moderate positive activity while the activity in the inner plexiform layer, the outer and inner nuclear layers and in the ganglion cell layer was negligible. The layer of nerve fibers and internal limiting membranes were completely negative. In the layer of neuroreceptors, the outer half of this layer gave a very strong positive reaction and the inner half a very mild positive reaction. The significance of this varied distribution in the neuroreceptor layer is discussed. A comparison of distribution of some important enzymes in the myelin sheaths of peripheral nerves, pigment epithelial cells of the retina, and the inner and outer segments of the neuroreceptor layer is made to show that each tissue is differently equipped with various enzymes, due probably to a fundamental difference in their functions.
NAPHTHOL AS-BI β-d-GLUCOSIDURONIC ACID; ITS SYNTHESIS AND SUITABILITY AS A SUBSTRATE FOR β-GLUCURONIDASE
Authors: FISHMAN WILLIAM H. , NAKAJIMA YASUO , ANSTISS C. , GREEN SIDNEY 
Published: April 2011

1. Naphthol AS-BI β-d-glucosiduronic acid has been prepared by first reacting potassiumnaphtholate AS-BI with acetobromomethyl glucuronate in absolute alcohol and then by removing the protecting acetyl and methyl groups with barium hydroxide and hydrochloric acid. A stable crystalline product gave the elemental and group analytical values expected for C24H22O9NBr·0.6 H2O and yielded the expected equimolar amounts of naphthol AS-BI and glucuronic acid on complete hydrolysis with purified rat liver β-glucuronidase. Contaminants were found absent after examining the product by thin layer chromatography. The glucuronide is stable at room temperature (20°C) as a solid or in aqueous solution. 2. Purified rat liver β-glucuronidase hydrolyzes the glucosiduronic acid at a rate somewhat more rapidly than it does phenolphthalein-, p-nitrophenyl- and 8-hydroxyquinoline β-d-glucosiduronic acids. The Michaelis constant is 0.000021 M, the optimum pH is 4.5. There is a linear relationship to enzyme concentration. Saccharolactone and the presence of a second substrate effectively inhibit the reaction. 3. Incidentally to the main objectives of the study, a simple colorimetric method was devised for measuring enzymically-liberated naphthol AS-BI in the kinetic studies. 4. It is concluded that naphthol AS-BI β-d-glucosiduronic acid is a suitable substrate for β-glucuronidase and that it lacks none of the desirable attributes of the classical biochemical and histochemical substrates. Moreover, it is now possible to undertake definitive studies on the design of enzymorphologic techniques for β-glucuronidase utilizing this new substrate.
THE REACTIONS OF SULFATED POLYSACCHARIDES TO SEVERAL HISTOCHEMICAL TESTS
Authors: YAMADA KAZUYORI 
Published: April 2011

1. The reactions of several sulfated polysaccharides and glycogen to a series of histochemical tests are described. 2. The accepted significance of a positive periodic acid Schiff reaction appears to hold true in these substances. 3. Although the alcian blue staining of the sulfated polysaccharides is attributed primarily to their acidic groups, their appropriate hydroxyl groups can influence the stainability. 4. The azure A metachromasia and methylene blue staining of the sulfated polysaccharides confirm largely the validity of the hitherto established reaction mechanisms. A finding worthy of recording is that chromic acid oxidation abolishes metachromasia of some sulfated polysaccharides. 5. Throughout the present study discussions were made on the possible mechanisms involved in the reactions observed.
THE HISTOCHEMICAL DEMONSTRATION OF PANCREATIC-LIKE LIPASE AND COMPARISON WITH THE DISTRIBUTION OF ESTERASE
Authors: ABE MUNEAKI , KRAMER STANLEY P. , SELIGMAN ARNOLD M. 
Published: April 2011

A new histochemical method for lipase was developed using naphthol-AS nonanoate as a substrate and sodium taurocholate as an activator. In a comparison with esterase, pancreatic-like lipase activity was studied in various tissues. As a result, it was disclosed that the pancreatic acinar cells show the highest activation with sodium taurocholate, and lipase activity is located in the region immediately surrounding the zymogen granules and not within the granules themselves. Maximum activity of esterase and lipase differed among various tissues of the rat. Esterase activity is most intense in liver and kidney followed by pancreas. Pancreatic-like lipase activity is most striking in pancreas, less so in the liver and in the kidney the activity is rather weak. Because of the exceptional esterase activity of macrophages, mast cells and pericytes of the cerebral capillaries, pancreatic-like lipase activity should be ascribed to them with caution and only in terms of the difference between reaction without and with sodium taurocholate.
CYTOCHEMISTRY OF PARACOCCIDIOIDES BRASILIENSIS. I. CYTOCHEMISTRY OF CYTOPLASMIC POLYSACCHARIDES IN YEAST FORM CULTURES WITH LIGHT MICROSCOPE
Authors: CARBONELL LUIS M. , CASTEJON HAYDEE , POLLAK LADISLAO 
Published: April 2011

1. The histochemical distribution and localization of cytoplasmic polysaccharides are studied in the yeast form of P. brasiliensis. Glycogen and an acid mucopolysaccharide probably containing sulfate groups are demonstrated. These polysaccharides disappear after the sixth day of culture. The glycogen and the acid mucopolysaccharide are bound to protein. 2. Stains and enzyme treatments applied in sequence help demonstrate the site of different polysaccharides.
QUANTITATIVE STUDIES ON ISOLATED PANCREATIC ISLETS OF MAMMALS: ADENOSINE TRIPHOSPHATASE ACTIVITY IN NORMAL AND OBESE-HYPERGLYCEMIC MICE
Authors: TÄLJEDAL INGE-BERT , HELLMAN BO , HELLERSTRÖM CLAES 
Published: April 2011

Microchemical and histochemical methods were used for the characterization, localization and assay of adenosine triphosphate (ATP) splitting enzymes in homogenates and sections of the endocrine pancreas from obese-hyperglycemic mice and their lean litter mates. The following observations were made: 1. Dephosphorylation of ATP was maximal at pH 9.1. It was strongly stimulated by magnesium ions at an optimal concentration of 1 mM. ATP cleavage was inhibited by adenosine diphosphate, sodium azide and p-chloromercuribenzoic acid. The addition of l-cysteine, sodium cyanide or sodium fluoride to the substrate medium had no effect on the enzyme activity. Substitution of ATP by equimolar amounts of other phosphate esters in the medium considerably reduced the substrate cleavage. These results are taken as evidence for the presence of sulfhydryl-dependent adenosine triphosphatase (ATPase) in the islet tissue. 2. Histochemical staining revealed a strong ATP splitting enzyme activity in the capillaries and walls of larger blood vessels throughout the pancreas; a rather weak and diffuse cytoplasmic reaction being found in the islet cells. 3. Microchemical assays revealed a lower cleavage of ATP in the islets as compared with the exocrine pancreas and the liver. The cleavage of ATP was more intense in the islets of the obese-hyperglycemic mice than in those of the lean litter mates. 4. Starvation for 7 days induced no significant changes in the enzyme activity of the endocrine pancreas.
EXTRACTION RESISTANCE OF SUDAN STAINED MAST CELLS AFTER PREVIOUS ACID TREATMENT
Authors: FEAGLER JOHN R. , MCMANUS J. F. A. 
Published: April 2011

1. Sudan black B alone stains the mast cells very faintly. The granules do not appear very distinct or numerous. 2. The periodic acid Schiff (PAS) technique stains the mast cells; however, the results are variable. When stained, the mast cell granules appear slightly darker than the cytoplasm. 3. When PAS and Sudan black are used together, the mast cell granules gain an avidity for the dye and are stained black. 4. Brief acid treatment prior to immersion in Sudan black results in granules gaining an avidity for the Sudan dye that is extraction resistant. The reaction appears dependent upon modifying the mast cell granules by acid treatment. It appears as though the hydrogen ion changes some physical or chemical property of the granules so that they gain an avidity for the dye. 5. Acetylated Sudan black shows an affinity for the mast cell granules greater than Sudan black alone. This is not increased by previous acid treatment. Acetylated Sudan black also stains the mast cell nucleus.
SYNTHESIS AND PROPERTIES OF MODEL SYSTEMS, WITH THEIR USE IN STUDYING THE SCHIFF REACTION IN HISTOCHEMISTRY
Authors: HARDONK M. J. , VAN DUIJN P. 
Published: April 2011

The synthesis of cellulose films containing several reactive groups is described. These films were developed for use as histochemical model systems to study several Schiff-type reactions. The synthesized model systems include cellulose membranes to which amino groups are covalently bound, cellulose membranes containing sulfhydryl groups, and cellulose membranes to which deoxyribonucleotides have been bound by means of their phosphate group. Evidence is given for the structure of these cellulose derivatives.
HISTOCHEMICAL STUDIES ON THE ROLE OF THE MAST CELL IN CALCERGY
Authors: SELYE HANS , GABBIANI GIULIO , SERAFIMOV NIKOLA 
Published: April 2011

Experiments were performed on rats, using special histochemical stains for the demonstration of metachromatic materials, lead, calcium, phosphate and carbonate, to study calcergy, the induction of soft-tissue calcification by topical treatment with certain compounds such as lead acetate. In simple calcergy, when lead acetate is directly injected into the subcutis, it is seen to impregnate the collagen fibers in the injection site without noteworthy participation of the mast cells. The lead-treated area secondarily attracts calcium, phosphate and possibly carbonate. In mastocalcergy, the lead acetate is injected intravenously and local calcification is produced by the simultaneous subcutaneous injection of a mast-cell discharger such as polymyxin. Here, the mast cells in the treated area first show degranulation without any mineralization, but this is soon followed by lead uptake on the mast-cell granules with secondary attraction of calcium, phosphate and possibly carbonate. Finally, the discharged and calcified mast-cell granules disintegrate into a dustlike, fine precipitate which is transferred to the circumjacent collagen fibers where it initiates an intense process of mineralization, eventually leading to the complete petrification of the polymyxin-injected area. Apparently, in addition to its already well established functions of histamine, heparin and serotonin production, the mast cell also plays a role in mineralization.
INTRACELLULAR MONOAMINE OXIDASE AND LIPIDS IN DIFFUSION CHAMBER CULTURES OF THE PINEAL ORGAN
Authors: O'STEEN W. KEITH , DILL RUSSELL E. 
Published: April 2011

Outgrowth of rat pineal tissue in diffusion chambers began with a migration of connective tissue cells from the implant, and this was followed by a spreading of the parenchymal cells to the margins of the chambers. Diffusion chamber implants and fresh slices of pineal organ were incubated for monoamine oxidase (MAO) in a medium containing nitro-blue tetrazolium and tryptamine hydrochloride. The MAO reaction was visualized as black formazan granules in parenchymal cells of all cultures throughout the entire 21 day observation period. These cytoplasmic granules appeared to increase in number and in size during the later time periods. A positive enzyme reaction also was present in slices of fresh pineal organ treated in the same medium and in a medium without the substrate, tryptamine hydrochloride. Pineal tissues with prior treatment in a MAO inhibitor gave a negative reaction. Some parenchymal cells in diffusion chamber implants accumulated many large lipid droplets. There were fewer of these cells than MAO positive cells, and they appeared to migrate from the implant at a slower rate.
IN VITRO BINDING OF NUCLEIC ACIDS TO TISSUE SECTIONS AFTER REMOVAL OF TISSUE NUCLEIC ACIDS
Authors: DAOUST R. 
Published: April 2011

Rat tissue sections were freed from nucleic acids by enzymatic or chemical extraction methods, immersed in solutions of nucleic acids from various sources and stained with toluidine blue. Tissue sections immersed in solutions of DNA showed intense nuclear and cytoplasmic staining; similar results were obtained with tissue sections placed in solutions of RNA. Thus both DNA and RNA can bind to nuclear and cytoplasmic sites in tissue sections freed from nucleic acids. The experiments indicated however that in vitro binding of nucleic acids to tissue sections was not specific to original sites of nucleic acid binding and the reactions showed no particular species or organ specificity.
DISTRIBUTION OF β-GLUCURONIDASE ACTIVITY IN RAT TISSUES EMPLOYING THE NAPHTHOL AS-BI GLUCURONIDE HEXAZONIUM PARAROSANILIN METHOD
Authors: HAYASHI MASANDO 
Published: April 2011

The histochemical distribution of β-glucuronidase activity in rat tissues as demonstrated by the naphthol AS-BI glucuronide and hexazonium pararosanilin method has been described. The present method has demonstrated a close correlation between the amount of stain derived from the β-glucuronidase reaction and the reported enzyme activity by homogenate assay in every tissue examined. Tissues known to be enzyme-rich by homogenate assay showed a rapid and intense staining in a great number of cells. These tissues were the preputial gland, spleen, liver, epididymis, ovary, lymph node, thymus, kidney and thyroid. In enzyme-poor tissues, the staining was negative or limited to a small number of specific cells. These tissues were the brain, spinal cord, eye, testis, and muscle. Cells responsible for the intense staining in all tissues examined were those of the macrophage system, parenchymal cells of liver and kidney and several types of mucous membrane cells. In each cell the major staining appeared as discrete and, most often, spherical granules in the cytoplasm, and no staining was observed in the nucleus, Size, number and location of the granules was quite variable depending on the type of cells. Large granules were especially abundant in macrophages. The possible significance of this enzyme distribution is discussed briefly.
ISOZYME HISTOCHEMISTRY: THE DISPLAY OF SELECTIVE LACTATE DEHYDROGENASE ISOZYMES IN SECTIONS OF SKELETAL MUSCLE
Authors: BRODY IRWIN A. , ENGEL W. KING 
Published: April 2011

The addition of chemical agents to the standard lactate dehydrogenase (LDH) incubating medium permits the histochemical display of selective LDH isozymes. Excess lactate decreases the intensity of the fast-moving isozymes, and urea decreases the intensity of the slow-moving isozymes. Application of these modified incubating media to histologic sections allows the display of selective LDH isozymes in unhomogenized tissues. Thus, predominance of the slow-moving LDH isozymes in guinea pig gastroenemius and of the fast-moving isozymes in guinea pig soleus were found on the electrophoretic patterns of the muscle extracts and confirmed on the intact tissues. The modified incubating media were used to localize specific LDH isozymes in normal human skeletal muscle. Ry extraction of the sections with saline or acetone the intracellular sites of isozyme activity were identified with recognized cell constituents. It was concluded that the slow-moving LDH isozymes predominate in the aqueous sarcoplasm of Type I fibers while the fast-moving isozymes predominate in the aqueous sarcoplasm of Type II fibers and in the mitochondria and lipid-mitochondrial complexes. The Pattern of dark and light fibers, seen after incubation in the standard LDH medium, was found to be due to the greater activity of the slow-moving isozymes in the Type I than the Type II fibers. Thus technique of isozyme histochemistry thus allows study of the intracellular localizations and the biochemical roles of specific LDH isozymes in skeletal muscles.
SOME ASPECTS OF THE SPECIFICITY OF FATTY ACID ESTERASES
Authors: HOFSTEE B. H. J. 
Published: April 2011

A comparison has been made of the meaning of the experimental kinetic constants Vm, KM and Vm/KM with respect to group transfer by a hydrolase proceeding either with or without obligatory intermediary transfer of the group to the enzyme. The results have been applied to the interpretation of kinetic data in a number of actual cases involving esterases whereby either several substrates have one of these experimental constants in common or the addition of a modifier has no effect on one of these constants with respect to a particular substrate.