bluuebunny/crossref_metadata_2025_split huggingface dataset:
THE APPLICABILITY OF A NEW SCHIFF BASE, GLYOXAL BIS (2-HYDROXYANIL),FOR THE CYTOCHEMICAL LOCALIZATION OF IONIC CALCIUM
Authors: KASHIWA HERBERT K. , ATKINSON WILLIAM B. 
Published: April 2011

A simple pocedure for localizing calcium ions in tissues and in cells with glyoxal bis (2-hydroxy-anil) (GBHA) has been described. The GBHA molecule, in alcoholic solution made alkaline with NaOH, chelated with Ca, Ba, Sr, Cd, Cu, Co, and Ni ions, forming colored precipitates. All the colored GBHA complexes, with the exception of the red Ca-GBHA granules, were decolorized when immersed in a solution containing carbonate and cyanide. Of the methods available for demonstrating calcium ions, the sensitivity of the GBHA method was comparable only to the Scott-Packer electron microscope method; the latter, however, localized calcium plus magnesium while the GBHA method was specific for calcium ions. In tissues that were fixed in 10% neutral formalin, alcohol-formalin or absolute alcohol, false localization due to binding of extraneous or diffused calcium ions to polyanionic sites was evident. The false localization of calcium ions was prevented by processing the tissues by the freeze-dry or the freeze-substitution method, dehydrating the tissue below the cutectic point of calcium chloride. The GBHA method was not able to demonstrate calcium ions present as insoluble salts, such as calcium carbonate or as apatite crystals, giving false negative results.
CYTOCHEMICAL STUDIES OF RABBIT AORTIC CELLS IN TISSUE CULTURE
Authors: KOIDE REIZO , POLLAK O. J. 
Published: April 2011

The enzymatic activities of acid and alkaline phosphatase, adenosine triphosphatase, carboxyl esterase and succinic dehydrogenase as well as the lipid, cholesterol and acid mucopolysaccharide content in three types of cells cultured from normal and cholesterol-fed rabbit aortae were studied. Although enzymatic activities varied from cell to cell in the same culture, there was good evidence that "endothelial" type III cells had more activity than the other two cell types. With normal aging enzymatic activity seemed to diminish in all three cell types. Giant cells were usually more active. Degenerated cells also had somewhat increased enzymatic activity. However, type I and II cells always had considerably less activity than type III cells, regardless of their origin, age or shape. Type III cells exhibited intense activity in acid phosphatase; moderate activity in esterase and in succinic dehydrogenase; and weak or almost negligible activity in alkaline phosphatase and adenosine triphosphatase. In "fibroblastic," type I or II cells all these enzymatic activities were weak or negligible. No differences of enzymatic activity were seen between cells from normal and atherosclerotic aorta but for the esterase activity which was highest in type III cells from plaques. With the exception of carboxyl esterase activity which seemed to be slightly higher in the type III cells from atherosclerotic aorta, cells of different origin did not differ in enzymatic activity. Some type I or II cells changed their shape and became round (9), but these round cells did not exhibit stronger enzymatic activities than the spindle forms. On the other hand, large size or differently shaped type III cells retained their greater activity. Thus, greater activity of type III cells was not due to shrinking of the cytoplasm and subsequent condensation of the cell. Studies of other enzyme systems are under way. Very preliminary results indicate that "endothelial" cells yield considerably higher (G. Nadi) cytochrome-c oxidase than other cells. "Endothelial" or type III cells contained more lipid and acid mucopolysaccharide than "intermediate" or type I cells and "fibroblastic" or type II cells. "Endothelial" our type III cells from atherosclerotic plaques contained cholesterol in their cytoplasm while none of the cells from normal aorta did. Type III cells are metabolically the most active cells cultured form the aortic wall. Their activity may play a major role in the development of the atherosclerotic plaques. Morphologic and functional characteristics of these cells led to the suggestion that they may be endothelial cells or, possibly, macrophages. The fact that only type III cells from atherosclerotic aorta contain much cholesterol and that they appear in larger numbers than cells from normal aorta is held supportive evidence that they are endothelial cells. Results with cells in tissue cultures were compared with reports on histochemical findings in aortae.
RELIABILITY OF THE RADIOAUTOGRAPHIC TECHNIQUE FOR THE DETECTION OF NEWLY SYNTHESIZED PROTEIN
Authors: DROZ B. , WARSHAWSKY H. 
Published: April 2011

It is widely believed that the radioactivity detected by radioautography after administration of a labelled amino acid is due to newly-synthesized protein. The validity of this postulate was examined in work done on the organs of mice injected with leucine-C14 and sacrificed one-half or 24 hours later. Pancreas, liver, kidney and cerebellum were removed and divided into 3 aliquots to determinse (a) the total radioactivity, (b) the biochemical distribution of the radioactivity in the fresh organs, (c) the losses of radioactivity occurring after Bouin fixation and in the course of histological processing as well as the biochemical distribution of the radioactivity retained in histological sections. In addition, an attempt was made to extract radioactivity from mounted sections by various reagents. It was found that 10-50% of the total radioactivity is lost in Bouin's fluid (presumably as free leucine) and in ethanol and dioxane (presumably as lipid-linked metabolites). As for the radioactivity persisting in the processed organs, only insignificant amounts were found in the nucleic acid fraction, while most of it was associated with protein. The possibility that the radioactive amino acids are bound to protein by ionic linkage was examined by extracting sections with strong acids and bases, but only insignificant amounts were thus lost. It is concluded that at least 91-97% of the radioactivity retained in histological sections after labelled amino acid injection is firmly bound to protein (presumably by peptide bonds). Thus, radioautography under these conditions is a reliable method to detect and trace newly-synthesized protein.
OSMIUM TETROXIDE FIXATION OF LIPIDS: NATURE OF THE REACTION PRODUCTS
Authors: RIEMERSMA JELLE C. 
Published: April 2011

To study the possible reaction mechanisms involved in osmium tetroxide fixation as used in electron microscopy, the products were analyzed which result from the interaction of osmium tetroxide and a number of pure substances. By means of paper chromatography, differences could be observed between the reaction products of 1-decene and oleic acid on the one side and of lecithin on the other. In the case of 1-decene and oleic acid, an intermediate osmic acid ester, as postulated by Criegee's reaction scheme, appears to be the main reaction product. In the case of lecithin, the reaction product is mainly a corresponding dihydroxy compound; here the intermediate osmic ester is readily split with formation of osmium dioxide and dihydroxy lecithin. The presence of a phosphoryl choline group as in lecithin thus appears to influence the course of the reaction with osmium tetroxide. If a polar group as well as double bonds play a part in the reactions, this might explains electron microscopic findings suggesting that during osmium fixation the deposition of osmium dioxide probably occurs at the polar ends of the lipid molecules. Our chromatographic experiments show that osmium dioxide, as a colloidally dispersed hydrate, is negatively charged and capable of Coulomb interaction with the quaternary ammonium group present in lecithin. Combined with the finding that osmium dioxide is readily split off from the osmium ester derivative of lecithin which is formed initially, this evidence strongly favors the hypothesis that osmium dioxide is deposited in the polar strata of lipid micelles.
THE OXIDATION OF LACTATE AND α-GLYCEROPHOSPHATE BY RED AND WHITE SKELETAL MUSCLE: II. HISTOCHEMICAL STUDIES
Authors: VAN WIJHE M. , BLANCHAER M. C. , JACYK W. R. 
Published: April 2011

The use of phenazine methosulfate (PMS) in the histochemical demonstration of lactate and α-glycerophosphate dehydrogenases in skeletal muscle has been examined. When PMS is added to the conventional dehydrogenase reagents, these enzymes appear to be at least as active in the white fibers as in the red. In the absence of PMS, the red fibers appear to be much richer than the white in lactate and α-glycerophosphate dehydrogenase. The conventional methods without PMS apparently give a false impression of the distribution of diphosphopyridine nucleotide (DPN)-linked dehydrogenases in skeletal muscle because of their dependence on the indigenous tissue level and intracellular localization of reduced DPN (DPNH)-diaphorase. A leakage of both enzymes from the sections into the histochemical dehydrogenase reagents occurred both in the presence and absence of PMS. When allowances are made for such losses, it appears that the PMS results are compatible with other evidence that the glycolytic dehydrogenases are more active in white than in red skeletal muscle fibers and that these enzymes are largely confined to the cytoplasm.
STUDIES OF THE CYTOCHEMISTRY OF NUCLEOPROTEINS III. DEMONSTRATION OF DEOXYRIBONUCLEIC-RIBONUCLEIC ACID COMPLEXES IN MAMMALIAN CELLS
Authors: LOVE ROBERT , RABOTTI GIANCARLO 
Published: April 2011

1. On the basis of susceptibility to digestion with dilute nucleases, the nine types of ribonucleoprotein (RNP) stainable by the toluidine blue-molybdate (TBM) method can be divided into four groups. The first group is resistant to the action of dilute ribonuclease (RNase) but is sensitive to deoxyribonuclease (DNase) and consists of both types of chromosomal RNP, perichromosomal RNP and intranuclear amorphous parachromatin. The second group is digested by dilute RNase and by DNase, and consists of the amorphous parachromatin that diffuses into the mitotic spindle and the nucleolini. The third group is digested by dilute RNase, but not by DNase and consists of both types of diffuse cytoplasmic and the granular cytoplasmic RNP. The pars amorpha of the nucleolus constitutes the fourth group which is resistant to digestion by dilute RNase and DNase. 2. Biochemical and other studies indicate that the effects of digestion by the enzymes is not due to contamination of RNase by DNase or vice versa. 3. Staining of chromatin by TBM (method C) is usually due to dye-binding by RNA and DNA and the intensity of staining is reduced by treatment with RNase. When the effective concentration of dye is low only RNA appears to be stained; digestion with RNase then leads to an increase in the intensity of staining. It is suggested that RNA may sterically mask the phosphoryl groups of DNA. A method for selectively staining RNA in the chromatin and chromosomes is described. 4. The observations suggest that the chromosomal, perichromosomal RNP and possibly the nucleolini and intranuclear parachromatin are present as a DNA-RNA hybrid. Current knowledge of the nature and enzyme susceptibility of informational RNA and its DNA hybrid suggest that the major component of the hybrid stained by TBM is informational RNA and that this type of RNA is present in the amorphous parachromatin of the mitotic spindle.
QUATERNARY AMMONIUM COMPOUNDS IN CONNECTIVE TISSUE HISTOCHEMISTRY: I. SELECTIVE UNBLOCKING
Authors: KELLY JOHN W. , BLOOM GUNNAR D. , SCOTT JOHN E. 
Published: April 2011

Certain features of an accepted biochemical system for fractionating biological polyanions were adapted for more precise identification of acid mucopolysaccharides in histological preparations. Tissue studies were based on two major steps: (1) complete blocking of all polyanions by a quaternary ammonium detergent and (2) selective unblocking by increasing concentrations of an electrolyte. Critical stages were visualized by suitable stains for acid mucopolysaccharides. Results with tissues and certain polyanion models showed that both acid mucopolysaccharide and nucleic acid structures behaved histochemically, under proper conditions, as predicted from the biochemical reactions of such polyanions as hyaluronic acid, chondroitin sulfate, heparin, deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). Chemical fixation involving formaldehyde seriously altered the expected positions of nucleic acid structures in the unblocking sequence; this was corrected by use of cetylpyridinium chloride in the fixing solution. Considering the biochemical background of these methods and several different histochemical applications already accomplished, numerous possibilities are open for extending the fundamental and practical uses of quaternary ammonium compounds in microscopic studies.
A QUANTITATIVE STUDY OF THE FEULGEN REACTION WITH THE AID OF HISTOCHEMICAL MODEL SYSTEMS
Authors: HARDONK M. J. , VAN DUIJN P. 
Published: April 2011

This paper reports some quantitative results with three histochemical model systems. The model systems are DNA-cellulose, deoxyadenylic acid-cellulose and deoxyguanylic acid-cellulose, and DNP-bovine albumin-polyacrylamide. By measuring the UV-absorption of the nucleotide-cellulose after various hydrolysis times, the rates of splitting-off of the purines by HCl were determined. Adenine is found to be split off twice as fast as guanine. The phosphorus/pararosaniline ratio is found to differ considerably from the theoretical value calculated on the assumption that every free aldehyde group reacts with pararosaniline. The differences in the values found for the phosphorus/pararosaniline ratio of the DNA-cellulose and the nucleotide-cellulose can be related to the differences in the absorption spectra of these models, which points to different degrees of substitution at the amino groups of pararosaniline. Addition of protein to DNP in a polyacrylamide film does not alter the phosphorus/pararosaniline ratio.
THE EFFECT OF MENADIONE AND PHENAZINE METHOSULFATE ON THE TETRAZOLIUM REDUCTION SYSTEM UNDER HISTOCHEMICAL CONDITIONS
Authors: HASHIMOTO T. , KALUZA J. S. , BURSTONE M. S. 
Published: April 2011

The influence of phenazine methosulfate and menadione on the activity of DPNH- and TPNH-tetrazolium reductase, succinic, lactic, and triphosphopyridine nucleotide-linked isocitric dehydrogenase were studied with reference to applied histochemistry. Fresh frozen sections of various tissues were employed in conjunction with nitro blue tetrazolium as the hydrogen acceptor. Phenazine methosulfate was found to produce a variable effect upon staining, enhancing it or suppressing it according to incubation conditions. Suppression was most pronounced with higher concentrations. Menadione had no suppressive influence and was useful in increasing tetrazolium reduction. Its usefulness is, however, limited unless a soluble factor, possibly DT-(DPNH-TPNH)-diaphorase, can be prevented from diffusing into the medium, and "nothing dehydrogenase" is selectively inactivated.
A HISTOCHEMICAL STUDY OF SUBSTRATE SPECIFICITY FOR THE STEROID 3β-ol DEHYDROGENASE AND ISOMERASE SYSTEMS IN HUMAN OVARY AND TESTIS
Authors: GOLDBERG B. , JONES G. E. S. , BORKOWF H. I. 
Published: April 2011

Either the enzyme which oxidizes 3β-ol groups in C-19 and C-21 steroids is properly designated a steroid-3β-hydroxy dehydrogenase or there is a multiplicity of such dehydrogenases. Although DHA is a satisfactory substrate for the human ovary, adrenal, and placenta, it is not generally so for the human testis. Pregnenolone is only at times a moderately satisfactory substrate in the ovary and is not satisfactory for the testis. In the present histochemical study two Δ4,3β-ol steroids have been reported as satisfactory substrates for human endocrine tissues as well as liver and intestine. These new substrates make it possible to demonstrate activity in human testicular tissues and at probable sites of androgen production. If these tenets hold, they indicate that it is more satisfactory to use unphysiologic substrates for the histochemical demonstration of enzymes as this will obviate the interplay of homeostatic mechanisms which must constantly be at work to preserve the cellular equilibrium. Although the data might suggest that the human testis, in contrast to the ovary, is deficient in isomerizing capacity and that this step precedes oxidation of the 3β-ol group, the biochemical data do not support this theory. A more probable explanation is that either DHA, its reaction product 5-androstenedione, or both, inhibit the human testicular enzyme system, while progesterone inhibits that of the corpus luteum; thus a difference between the dehydrogenases of these two human tissues appears to exist.
CHANGES IN HYDROLASE ENZYMORPHOLOGY OF RAT UTERUS AND VAGINA AFTER ESTROGENS; NAPHTHOL AS SUBSTRATES
Authors: WATANABE KEIICHI , FISHMAN WILLIAM H. 
Published: April 2011

The early enzymorphologic changes of rat uterus and vagina on castration and following estrogen administration (estradiol-17β, estrone and estriol) were examined with regard to alkaline phosphatase, esterase, β-glucuronidase and acid phosphatase using naphthol AS compounds as substrates. Castration caused a marked decrease of enzyme activities except for alkaline phosphatase of capillary endothelium and for nonspecific esterase. All activities, compared to castrated controls, were markedly increased by estrogen administration except for the alkaline phosphatase of capillary endothelia. Estriol and estrone were more potent than estradiol-17β in the early stage of estrogen treatment (4-hour interval). The response of alkaline phosphatase of striated border of lumenal uterine epithelium to estrogen treatment is unique in that the other hydrolases were absent from this site. The estrogen sensitivity of the hydrolases of connective tissue cells of rat uterine stroma is clearly evident for the first time. Of interest also is a comparison of β-glucuronidase and acid phosphatase in which naphthol AS-BI is the product of enzyme hydrolysis in each case. Although the intensity of the respective staining reactions correlated well with each other in relation to the events of castration and estrogen administration, there were clear differences evident. Finally, the new findings demonstrate the value of the improved techniques as well as resolve some ambiguities of earlier findings obtained with diverse methods.
ANALYTICAL CYTOLOGY OF MUCIN PRODUCTION. V. A NEW MUCINOUS TISSUE CULTURE LINE
Authors: ROSAN ROBERT C. , KERRIGAN JUDITH A. 
Published: April 2011

1. Morphologic and cytologic detail is presented on a hitherto unreported tissue culture line. 2. The cultures, which are derived from a biologically stabilized transplantable homologous mucinous tumor of hamster, display many of the features of the donor tumor. They elaborate mucinous materials, some of which accept the periodic acid Schiff stain and others of which take the Alcian blue stain. 3. The fluorescent periodic acid Schiff reaction identifies structures not generally considered to be Schiff positive following classical oxidation and/or blockade procedures. 4. In these cells, the nucleolus appears to have a definite envelope. It appears to complex with an acridine dye at pH 4 and gives reactions for aldehyde and ethylene groups.
A HISTOCHEMICAL ANALYSIS OF DEHYDROGENASE VARIETY IN THE DIFFERENT TYPES OF MUSCULAR TISSUES
Authors: DICULESCO I. , ONICESCO D. , MISCHIU L. 
Published: April 2011

Histochemically, we examined several types of dehydrogenases in the myoblasts from "in vitro" cultures of the chick embryos, in the gastroenemius of frog, in the pigeon pectoralis, in the heart of several animal species and in the myometrium of the rabbit in different functional states (non-gestatory, gestatory and postpartum In the myoblasts were active: succinic, isocitric, malic dehydrogenases, di- and triphosphopyridine nucleotide diaphorases, glutamic, α-glycerophosphate and lactic dehydrogenases and lactic acid, the two dehydrogenases of the pentose cycle and β-hydroxybutyric dehydrogenase. The cardiac muscular tissues and red striated muscle show high and very high reactions for the enzymes of the Krebs cycle and the diaphorases, and negative reactions for the rest of the studied enzymes. The white striated muscular tissue presents all the reactions, except those of the enzymes of the hexose shunt. This type of tissue is marked by a high enzyme activity of the anaerobic glycolysis. The gestatory and non-gestatory myometrium shows all the enzymatic reactions except lactic dehydrogenase. In the myometrium of the postpartum period one notes an enzyme decrease of the Krebs cycle, of the pentose cycle and of the lipidic metabolism and an intensification of the anaerobic glycolysis and the glucose dehydrogenase. Alcohol dehydrogenase was negative in all tissues studied with the exception of one weak reaction in the myometrium during the first postpartum hours.
HISTOCHEMICAL STUDY OF OXIDATIVE ENZYMES IN VERTEBRATE MUSCLES
Authors: OGATA TAKURO , MORI MASAHIKO 
Published: April 2011

A histochemical study was carried out on oxidative enzymes, succinic dehydrogenase, diphosphopyridine nucleotide (DPN)-dependent dehydrogenases (lactic, malic, α-glycerophosphate, glutamic, and β-hydroxybutyric dehydrogenases), DPNH-diaphorase, triphosphopyridine nucleotide (TPN)-dependent dehydrogenases (isocitrate, and glucose-6-phosphate dehydrogenase) and TPNH-diaphorase on various kinds of vertebrate muscles. The results obtained were as follows: 1. Most of the mammalian striated muscle fibers fell into three types following a histochemical demonstration of oxidative enzyme reaction: Small red muscle fibers showed higher activities of oxidative enzymes, while large white fibers lower activities and the third type of fibers, "medium fiber" which were intermediate in size showed activities between red and white muscle fibers. 2. The intrafusal fibers of mammalian muscle spindles are classified into three types by oxidative enzyme reactions, i.e., red intrafusal fibers, medium intrafusal fibers and white intrafusal fibers. 3. Regarding birds, chicken muscle was composed of three types of fibers, while pigeon and lovebird muscles displayed two types of fibers. 4. Reptile and amphibian muscle fibers were classed as small red fibers showing higher activities, medium fibers moderate activities and white fibers lower activities. 5. Fish red muscle was composed only of red fibers, and the white muscle of white fibers. The transitional part of red and white muscles, revealed three types of fibers. 6. Generally, succinic dehydrogenase, lactic dehydrogenase, malic dehydrogenase, α-glycerophosphate dehydrogenase showed a higher activity, DPNH-diaphorase, TPNH-diaphorase a moderate activity, glutamic dehydrogenase, isocitrate dehydrogenase a lower activity and β-hydroxybutyric dehydrogenase and glucose-6-phosphate dehydrogenase a trace or nonactivity in most vertebrate striated muscles.
HISTOCHEMICAL STUDY OF OXIDATIVE ENZYMES IN INVERTEBRATE MUSCLES
Authors: OGATA TAKURO , MORI MASAHIKO 
Published: April 2011

A histochemical study of oxidative enzymes was carried out on various kinds of invertebrate muscles and the results obtained were as follows: 1. Insect flight muscles showed the presence of the most intense activity of succinic dehydrogenase and high activity of α-glycerophosphate and glutamic dehydrogenases. 2. Regarding crustacean striated muscles, prawn muscle was distinguished into types of fibers by oxidative enzyme reactions. 3. In molluscan muscles, the squid and octopus muscle fibers were distinguished into two types by their oxidative enzyme activities. 4. The earthworm body wall muscle was made of only one type of fibers, whose oxidative enzyme activities were low.