bluuebunny/crossref_metadata_2025_split huggingface dataset:
HISTOCHEMICAL STUDY OF THE PIGMENT FOUND IN THE EPITHELIUM OF THE KIDNEY OF THE PHILODRY AS SCHOTTII (SCHLEGEL)
Authors: DA SILVA SASSO WILSON , DE CASTRO NYLCEO MARQUES , PACHECO IRINEO PONTES 
Published: April 2011

The authors studied histochemically the pigment observed within the cells of the proximal convolutions of the kidneys of the ophidian ( Philodryas schottii), known in Brazil as "parelheira". The results demonstrate the existence of two pigments: an argentaffin, found in greater quantity and a chromolipoid of the lipofuscin type, in smaller amounts.
THE EFFECT OF PHOSPHOMOLYBDIC ACID ON THE STAINABILITY OF CONNECTIVE TISSUES BY VARIOUS DYES
Authors: PUCHTLER HOLDE , ISLER H. 
Published: April 2011

The mechanism of the selective staining of connective tissue obtained in trichrome methods (e.g. Masson, Heidenhain, Goldner) by "mordanting" with phosphomolybdic acid was investigated in sections of rat intestine. Information on the distribution of phosphomolybdic acid was obtained by treating sections with this acid and then with stannous chloride. The molybdenum blue thus obtained was located in those very structures which were stained selectively in trichrome methods. The chemical characteristics of the dyes taken up after phosphomolybdic acid treatment were investigated by comparing sections stained with acid, basic and amphoteric dyes. In the absence of phosphomolybdic acid, only dyes with acidic groups yielded a staining of connective tissue, but this staining was not selective. In sections pretreated with phosphomolybdic acid, only dyes with basic groups stained connective tissue, and only dyes with strong basic groups yielded a selective staining of these structures. Basic and amphoteric dyes applied after phosphomolybdic acid had the same distribution as this acid. It is concluded that the polyvalent phosphomolybdic acid is forming a bridge between the substrate and the dye, holding the dye and probably also the substrate by their cationic groups.
PHYSICAL AND CHEMICAL PROTECTION OF CELL CONSTITUENTS AND THE PRECISE LOCALIZATION OF ENZYMES
Authors: SCARPELLI D. G. , PEARSE A. G. E. 
Published: April 2011

1. Physical and chemical protection of cytoplasmic constituents in tissue sections from the time of tissue removal until completion of the histochemical reaction resulted in a more precise intracellular localization of various enzymes. 2. In many instances sites of enzymatic activity were localized on rod-shaped and ovoid bodies resembling mitochondria and lysosomes. 3. Thermal, osmotic and chemical injury of cytoplasmic structures produced extensive intracellular and extracellular diffusion of enzyme and reaction product. 4. Once thermal damage had occurred mitochondrial swelling and enzyme diffusion appeared despite osmotic and chemical protection.
THE SITE OF A DIFFUSIBLE NUCLEOTIDE REVEALED BY A COMPARISON OF CHEMICALLY FIXED TISSUE WITH FROZEN AND DRIED TISSUE
Authors: BELL L. G. E. 
Published: April 2011

1. Using unfixed frozen and dried tissue as a standard it is shown that acid alcohol fixation extracts a diffusible nucleotide from the vicinity of the chromosomes. 2. It is shown that the denuded area contains protein and that the end groups are not available for reaction with protein reagents.
EVALUATION OF INDOPHENOL DERIVATIVES AS REAGENTS FOR DEMONSTRATING CYTOCHROME OXIDASE IN TISSUE SECTIONS
Authors: CRAWFORD DAVID T. , NACHLAS MARVIN M. 
Published: April 2011

Seven redox dyes were investigated as possible histochemical indicators of cytochrome oxidase activity. All failed to meet one prime requisite, namely, that the oxidized product remain at the site of enzymatic action. The "color fading" phenomenon was studied. This effect was one in which sections and homogenates became colored as a result of oxidase activity and then faded during further incubation. Evidence was provided that this reaction occurred as a result of endogenous dehydrogenase activity, mainly succinic dehydrogenase. The decoloration in solution could be prevented by N-ethyl maleimide, without inhibiting cytochrome oxidase.
HISTOCHEMICAL DEMONSTRATION OF ACID PHOSPHATASE ACTIVITY IN OSTEOCLASTS
Authors: BURSTONE M. S. 
Published: April 2011

High acid phosphatase activity was observed in osteoclasts of several species using a reproducible azo-dye technique. High activity of two distinct enzymes, acid and alkaline phosphatase, are associated with osteoclasts and osteoblasts respectivey. Althouth frozen-dried tissues are recommended for definitive studies, the enzyme techniques used give satisfactory results with cold acetone-fixed tissues. The most accurate localization of acid phosphatase in osteoclasts in controlled comparative studies is obtained with double-embedded frozen-dried undecalcified tissues in conjunction with naphthol AS-phosphates.
SAPONIFICATION AS A MEANS OF SELECTIVELY REVERSING THE METHYLATION BLOCKADE OF TISSUE BASOPHILIA
Authors: SPICER S. S. , LILLIE R. D. 
Published: April 2011

Saponification reverses the methylation blockade of cartilage and mast cell basophilia demonstrated by azure A above pH 3-4, but fails to do so at pH levels below this range. Saponification of oxidized hair cortex restores the methylation blocked basophilia demonstrable at pH 2 or higher. Mouse amyloid is characterized by a weak basophilia which is relatively resistant to methylation. The results are interpreted as indicating that methylation hydrolyzes the sulfate esters and esterifies other acid groupings in tissue sections.
A METHOD FOR THE CYTOCHEMICAL DEMONSTRATION OF SUCCINIC DEHYDROGENASE IN HUMAN LEUCOCYTES
Authors: DE SOUZA E. J. , KOTHARE S. N. 
Published: April 2011

1. A method for demonstrating succinic dehydrogenase in human leucocytes using a p-nitrophenyl substituted ditetrazole is described. 2. In some of the leucocytes the formazan appeared as fine blue discrete granules or rods.
EFFECTS OF ALLOXAN AND REDUCED FOOD INTAKE ON LIVER DEOXYRIBOSE NUCLEIC ACID OF THE RAT
Authors: DI STEFANO HENRY S. , DIERMEIER HAROLD F. 
Published: April 2011

Effects of alloxan, reduced food intake, starvation and biopsy procedure on the deoxyribose nucleic acid (DNA) content of rat liver tetraploid nuclei have been investigated. It was found that: (1) Alloxan treatment resulted in an increased amount of DNA. (2) Reduced food intake of treated animals Played no part in this increase. (3) Complete starvation for a period of six days did not effect the amount of DNA in these nuclei. (4) An increased amount of DNA was found in tetraploid nuclei of rat liver 13 days after removal of a small biopsy sample. The increased DNA in tetraploid liver nuclei of alloxan diabetic rats may be explained on the basis of regeneration of liver tissue due to some submicroscopic liver damage caused by the drug.
A SCHIFF REAGENT OF CALIBRATED SENSITIVITY
Authors: ELFTMAN HERBERT 
Published: April 2011

The sensitivity of Schiff reagent for the detection of low concentrations of aldehyde increases progressively as the concentration of sulfurous acid in the reagent is lowered. Calibration of sensitivity is readily accomplished by titrating the Schiff reagent against the iodine of Lugol's solution. Stability of Schiff's solution depends on decreasing the evaporation of sulfur dioxide and of retarding the change of sulfite to sulfate. For practical purposes these ends can be achieved by storage in the refrigerator of a solution with greater sulfurous acid content and acidity than the solution used for staining. Evacuation of the excess sulfur dioxide or mixing in proper proportions with used reagent allows the rapid production of a solution of the desired sensitivity. If chemical preservatives are required, 0.5% hydroquinone gives very good results. Spectrophotometric measurement of the colored solutions produced by adding aldehyde to Schiff reagent indicates that at least two distinct compounds may result. The one that is usually mentioned as characteristic of a positive response of aldehyde to the Schiff reagent has a violet color. When the concentration of aldehyde is so low that only a Schiff solution of low sulfurous acid titer will react with it, a red color is produced. Spectrophotometric analysis shows this compound to be different from the original basic fuchsin and validates its use as an endpoint in the reaction of aldehyde with Schiff's reagent.
CHOLINESTERASES IN THE MAMMALIAN TONGUE
Authors: ELLIS RICHARD A. 
Published: April 2011

Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) have been demonstrated histochemically in the tongues of five Primates and the rabbit. Sensory nerve endings in the tongue, including the subepithelial nerve net, Meissner's corpuscles and the end bulbs of Kraus, contain both AChE and BChE. The intraepithelial nerve endings and the taste buds in the foliate, vallate and fungiform papillae react for AChE only. The postganglionic parasympathetic nerve fibers innervating both the pure mucous and the pure serous glands of the tongue have abundant AChE. In addition to the AChE and BChE activity in their motor end plates, the skeletal muscle fibers of the tongue also have a concentration of cholinesterases at their point of insertion on the lingual fascia which is designated as the cholinesterasic apparatus. The Winkelmann and Schmit silver technique for nerve axoplasm is found to be unreliable for visualizing autonomic nerves.
ENZYMATIC HISTOCHEMISTRY OF ETHIONINE INDUCED LIVER CIRRHOSIS AND HEPATOMA
Authors: WACHSTEIN MAX , MEISEL ELIZABETH 
Published: April 2011

A number of enzymatic histochemical staining reactions were applied to the livers of rats that had been given dl-ethionine for periods up to 330 days. The ethionine lesion is characterized by early central hepatocellular necrosis, proliferation of oval cells, and degenerative and regenerative alterations in the liver cells leading to cirrhosis and areas of adenofibrosis. Finally, liver cell carcinoma develops. The zonal differences of various enzymatic staining reactions in the liver lobules characteristic for the normal liver disappear in regenerating liver lobules. Most enzymes are demonstrable in these regenerating cells, as for instance succinic dehydrogenase, nonspecific esterase, glucose-6-phosphatase, diphosphopyridine nucleotide diaphorase and others. Hepatic carcinoma cells likewise retain many of the staining reactions. There is, however, a complete lack of bile canalicular activity, which is best seen with the ATPase technique. Such a deficiency occurs also in some regenerating nodules without obvious evidence of malignancy. Thus, the lack of bile canalicular staining in cirrhotic liver lobules may indicate impending carcinomatous transformation. Oval "ductular" cells showed a lack of various enzymatic staining reactions, but did show moderate DPN diaphorase and nonspecific esterase activity. The origin of these cells from liver cells, although not proven, is definitely suggested. In areas of oval cell proliferation, there was marked increase in the intensity of sinusoidal staining with various phosphatase techniques. Fibrous tissue, particularly in areas of adenofibrosis, showed areas of marked adenosine triphosphatase activity. On the other hand, 5-nucleotidase activity was increased strongly in areas where reticulum fibers had been deposited. Increase in nonspecific esterase and acid phosphatase in Kupffer cells indicated apparently a marked stimulation of the reticuloendothelial system.
SOME OBSERVATIONS ON THE HISTOLOGY AND HISTOCHEMISTRY OF THE CHROMAFFIN CELLS PROBABLY STORING DOPAMINE
Authors: FALCK BENGT , HILLARP NILS-ÅKE , TORP ALF 
Published: April 2011

The presence of a new type of chromaffin cell in certain tissues in ruminants was reported in previous studies. The results obtained by indirect methods indicated that the cells store dopamine and that this amine is responsible for their chromaffinity. The concentrations of adrenaline, noradrenaline, 3,4-dihydroxyphenylalanine and 5-hydroxytryptamine were found to be too low to account for the chromaffin reaction. This paper is concerned with the general characteristics and the histochemical behaviour of the cells (for simplicity the cells are called dopamine cells). The dopamine cells contain cytoplasmic granules which may be selectively stained by a nuclear fast red staining method. By use of this method they are clearly distinguishable from other mammalian chromaffin cells. In all essential respects the dopamine cells resemble the catechol amine containing cells in the adrenal medulla. Their cytoplasmic granules show a direct chromaffin reaction with bichromate but no reaction after pretreatment with formalin. The formalin-fixed cells are argentaffin and strongly reduce ferricyanide, and retain their reducing capacity at least partly even after pretreatment with bichromate. The azo-coupling reaction with diazo-safranin is negative. The dopamine cells are clearly distinguished from the enterochromaffin cells by the chromaffin and azo-coupling reactions. The most significant result is that the dopamine and noradrenaline cells, after fixation in formalin or bichromate, have the same histochemical behavior. Since dopamine is chemically closely related to noradrenaline, this is what should be expected if the specific chromaffin cells in ruminant tissues contain dopamine. It is concluded that the results provide further support for the view that the specific chromaffin cells in ruminants store dopamine.
TETRAZOLIUM STUDIES ON THE RETINA: I. INTRODUCTION AND TECHNIQUE
Authors: KUWABARA TOICHIRO , COGAN DAVID G. 
Published: April 2011

The retina has physical dimensions that make it especially favorable for incubation studies in its unsectioned state. With blue tetrazolium anti suitable substrates, dehydrogenase activity is demonstrated in various, but characteristic, forms and places. A red color that depends on enzymatic activity is found in the reticular layers, nerve fiber layer, and lipid portions of the retina. This appears to represent a solubilization of reduced and partially reduced formazan in the lipid portions of the retina. The blue precipitate is found in mitochondria-like granules within certain cells, notably in the ellipsoids of the rods and cones, glial cells, Müller's cells, and to some extent in ganglion cells. It is also found as amorphous and rod-like crystalline deposits in the inner layers of the retina and as a fine dust-like deposit in the nerve fiber layer. Metabolic activity in the retina as reflected by tetrazolium precipitation persists for hours after death and is present about equally in peripheral and central regions of the retina. The reduction brought about by the utilization of specific substrates representing intermediates in the major metabolic pathways will be the subject of subsequent papers of this series.
ENZYMES CATALYZING SEQUENTIAL REACTIONS IN MOUSE BRAIN AND LIVER SUPERNATANT FRACTIONS: II. THE USE OF PHENAZINE METHOSULFATE
Authors: SHOWACRE JANE , DU BUY HERMAN G. 
Published: April 2011

1 . Spectrophotometric and manometric studies of pertinent non-enzymic solutions showed that reduced diphosphopyridine nucleotide is rapidly oxidized by phenazine methosulfate. The reduced phenazine methosulfate (PMS) is gradually oxidized by molecular oxygen. H2O2 is formed. A portion of this H2O2 is spontaneously split into H2O and O2 2. In mouse brain and liver supernatants, with PMS as dye and reduced diphosphopyridine nucleotide (DPNH) as substrate, the same reactions occur. In addition, the formed H2O2 is split by catalase demonstrated to be present in the supernatants used. Th oxygen taken up was a direct measure of the DPNH oxidized. 3. Pyruvate was the main competing electron acceptor studied. When it was added to, or formed by the supernatants, the oxygen uptake during DPNH-oxidation through PMS was reduced, depending on the relative concentration of the electron transferring systems present. In that case the amount of oxygen consumed, when added to the amount of lactate accumulated, was a direct measure of the DPNH oxidized. 4. When oxidized diphosphopyridine nucleotide (DPN) and an appropriate substrate were added to supernatants, the reoxidation by PMS of the DPNH formed by the DPN-requiring dehydrogenase components of a given enzyme group was measured. The oxygen uptake was a measure of the amount of DPNH formed, and thus a measure of the enzymic breakdown of the substrate by the enzyme system under study. 5. When the metabolism of a substrate by the enzyme system under study included the formation of pyruvate, the PMS-dependent oxygen uptake, added to the lactate accumulated, was a measure of the activity of the enzyme system. 6. Homogenized brain tissue yielded a supernatant with fragmented enzyme groups lacking functional flavoprotein. By contrast, as shown previously, this tissue yielded a mitochondrial fraction which retained all of the enzynes necessary for an integrated glucose-metabolizing enzyme system. 7. Homogenized liver tissue yielded a supernatant with flavoprotein linked enzyme systems less fragmented than those found in brain supernatants. At the same time it yields a mitochondrial fraction with an incomplete glycolytic enzyme system. 8. The data support the thesis proposed previously, that the enzymes found in the supernatant fraction of brain and liver cells represent mainly enzymes washed from the mitochondria, and therefore complementary to those retained by these organelles.
DISTRIBUTION OF PHOSPHORYLASE IN THE UTERI OF CYCLIC AND HORMONE TREATED RATS
Authors: BO WALTER J. 
Published: April 2011

Following the technic of Takeuchi and Kuriaki, phosphorylase activity was identified in the uterus of cyclic and hormone treated rats. In the proestrous uterus phosphorylase activity was more pronounced in the outer longitudinal muscle layer than in the inner circular layer. In the diestrous as well as in the castrate uterus, enzymatic activity was observed in the outer longitudinal muscle layer, and only scattered amounts in the inner circular layer. Following estrogen treatment phosphoryhase activity was very pronounced in both layers, particularly in the outer longitudinal layer. In castrate progesterone-treated rats enzymatic activity was observed in the outer longitudinal layer. In rats that received estrogen and progesterone concurrently, phosphorylase activity was heavy in the outer longitudinal muscle layer and weak in the inner circular muscle layer. In general, phosphorylase activity in the uterus of the rat corresponds to the distribution of glycogen in the rat uterus. However, enzymatic activity was observed in the uterus when glycogen was not detected by using the periodic acid-Schiff technic. The distribution of glycogen in the rat uterus is discussed.
A CORRELATIVE STUDY OF THE HISTOCHEMICAL PROPERTIES OF RODENT ACID MUCOPOLYSACCHARIDES
Authors: SPICER S. S. 
Published: April 2011

The staining properties of several rodent mucopolysaccharides with alcian blue and with azure A at controlled pH have been surveyed and compared with available radioautographic data of S35O4 incorporation in mucins. The effects of treatment with acid methanol and of sulfation on the staining have been determined. One sulfated, metachromatic and strongly acid type of mucin stains weakly if at all with alcian blue, remaining unstained by alcian blue in the combined alcian blue periodic acid Schiff or alcian blue-azure A (or safranin) procedures, but loses azurophilia while gaining alcian blue reactivity after mild (4 hr 37°) treatment with acid methanol. Some (probably) sulfated mucins stain fully with both dyes and others, presumably also sulfated, color only partially with azure A at pH 1.5, but maximally with alcian blue. The latter two types of substances partially or completely resist supression of either staining reaction by the "mild methylation". A fourth category stains meta- or orthochromatically at pH 2.5 to 3, or not at all, with azure A, and colors strongly with alcian blue. Some of the latter mucins are sulfated and remain unaltered after mild treatment with acid methanol, whereas others appear to be nonsulfated and lose basophilia reversibly with such treatment. It is suggested that alcian blue staining and metachromasia or orthochromasia with azure A above pH 2.5 which is blocked after 4 hours in acid methanol at 37°C and restored by brief saponification, may be attributed to nonsulfated mucopolysaccharides containing strongly acidic carboxyls. A four hour acidic methylation at 69° eliminates nearly all the basophilia of acid mucins, and saponification restores alcian blue staining to some, but not azurophilia below pH 3.0. Sulfation of histologic sections renders many neutral and all acid mucins fully chromotropic toward azure A at pH 1.5, but obstructs their alcian blue staining. Exposure to acid methanol after sulfation restores reactivity toward alcian blue and eliminates most artificial azurophilia; but the neutral mucins in Brunner's glands and mucous neck cells of rats and mice remain metachromatic and develop alcian blue affinity. Strong treatment with acid methanol increases the periodic acid Schiff staining of several subfated mucopolysaccharides of the rabbit. Staining with azure A or safranin after alcian blue differentiates alcian blue-negative and -positive sulfated polysaccharides. Failure of sulfate-free acid mucins to stain with alcian blue above the pH of their dissociation and resistance of the dye complex with these mucins to dissolution by acid or alkali is interpreted as evidence against the alcian blue-substrate complex involving a salt linkage.
THE LOCALIZATION OF RIBONUCLEASE ACTIVITY IN TISSUE SECTIONS
Authors: DAOUST R. , AMANO HARUKO 
Published: April 2011

The substrate film method was applied to localize ribonuclease activity in tissue sections and the distribution of the enzyme in the small intestine, large intestine, pancreas, thyroid, liver and skin of the rat was described. The resolution of the substrate film method has been improved in the present work by using a technique of film preparation which permitted us to obtain thinner films of substrate.
STUDIES ON THE HISTOCHEMICAL DIFFERENTIATION OF ENZYMES HYDROLYZING ADENOSINE TRIPHOSPHATE
Authors: FREIMAN DAVID G. , KAPLAN NANCY 
Published: April 2011

At least 3 enzymes or groups of enzymes present in dog kidney and jejunum are capable of splitting adenosine triphosphate at pH 9.4. In addition to nonspecific alkaline phosphatase and what appears to be a true adenosine triphosphatase, an enzyme behaving like a less specific organic polyphosphatase is present in small blood vessels and smooth muscle. These enzymes can be readily differentiated by various inhibitors such as cysteine, ethylenediamine tetraacetic acid, 80% ethanol, pretreatment in acetate buffer at pH 4.0 with or without added chloride or nitrate ion, or by incubation at 4°C. Differential effects can also be obtained by pretreatment with lead or zinc ion at pH 6.0. Of the substrates tested only inosine triphosphate gives results comparable with adenosine triphosphate. All other substrates, including various nucleotides and thiamine pyrophosphate, produce patterns similar to that obtained with adenosine diphosphate, or with adenosine triphosphate after pretreatment with buffer at pH 4.0. These observations suggest that an enzyme less specific than adenosine triphosphatase is being demonstrated. The similar patterns obtained following incubation in lead-adenosine triphosphate substrate at pH 7.2 also indicate that true adenosine triphosphatase cannot be shown by this method. 5'-Nucleotidase plays little role in these reactions under the experimental conditions used. Enzymes behaving in similar fashion have been noted in some human tissues. Of particular interest is a broad inner zone of the circular muscle coat in the small intestine of the dog which is strongly positive for adenosine triphosphatase and polyphosphatase, but negative for 5'-nucleotidase. A similar, but much narrower zone has been noted in the ileum and colon of man.