bluuebunny/crossref_metadata_2025_split huggingface dataset:
TETRAZOLIUM STUDIES ON THE RETINA: II. SUBSTRATE DEPENDENT PATTERNS
Authors: COGAN DAVID G. , KUWABARA TOICHIRO 
Published: April 2011

Unsectioned retinas were incubated in a medium consisting of buffer, blue tetrazolium and one of the following substrates: lactate, lactate-DPN, diphosphopyridine nucleotide (DPN), DPNH, and succinate. Control retinas incubated without substrate showed only an occasional reduction of tetrazolium and this was so slight as to be inconsequential. With lactate as the sole substrate no appreciable reduction of tetrazolium occurred at neutral pH but when DPN was also added a constant pattern of precipitation occurred that was qualitatively the same in all species. This pattern, associated with lactate-DPN substrate, was characterized by precipitation of tetrazolium in ganglion cells, Müller's fibers, and other glial cells. It was particularly obvious in human retinas and most particularly in pathologic retinas which were accompanied by gliosis. Also characteristic was the absence of activity in the ellipsoids of the rods and cones. The lactate-DPN activity was somewhat enhanced by incubation anaerobically or with cyanide. It was completely inhibited by p-chloromercuribenzoate or by borate. It was not affected by prior blinding of the animal with iodoacetate, and section of the optic nerve affected it only by abolishing the activity in the ganglion cells. When DPNH was used as a substrate the reduction of tetrazolium followed the same pattern. The inference is that lactate contributes the electron to DPNH and that tetrazolium reduction is effected through diaphorase. On the other hand lactate activity occurs at an alkaline pH (8.8-9.2) without the addition of DPN. In contrast to the lactate-DPN system, succinate substrates showed predominant activity in the ellipsoids of the rods and cones and in inner portion of the retina unassociated with glial cells or ganglion cells. It was approximately equal in all species. The succinate activity was greatly decreased in pathologic retinas associated with gliosis. It was inhibited by cyanide and malonate but not by borate. Section of the optic nerve caused no appreciable alteration but prior blinding of the animal with iodoacetic acid showed a loss of activity from the rods and cones and a significant increase in activity in the inner retinal layers. It is concluded that tetrazolium precipitation in the retina with lactate-DPN as a substrate is most marked in the glial cells and ganglion cells whereas that with succinate as a substrate is most marked in the nerve fibers and in the ellipsoids of the rods and cones.
THE ACTIVITY OF SUCCINIC DEHYDROGENASE IN THE EXPERIMENTAL EPENDYMOMA OF C3H MICE
Authors: OGAWA KAZUO , ZIMMERMAN H. M. 
Published: April 2011

1. The activity of succinic dehydrogenase in the transplantable ependymoma induced with methylcholanthrene in C3H mice was studied by both biochemical (QO2) and histochemical (using nitroneotetrazolium chloride) methods under aerobic conditions. The mitochondria were identified by Mallory's method in fixed tissue. 2. Succinic dehydrogenase activity in the ependymoma was much lower than in normal cerebrum. 3. In the normal cerebrum as well as in ependymoma there was close parallelism between the number, distribution and localization of mitochondria and succinic dehydrogenase activity. 4. In the normal cerebrum, neurons revealed the highest succinic dehydrogenase activity, followed by choroidal epithelium and ependymal cells. Formazan granules were scattered throughout the neuropil. Oligodendrocytes, particularly satellites, showed moderate activity. Occasionally very low activity was present in astroglia. 5. Ependymoma cells contained a smaller number of formazan granules than average normal cerebrum, and the mitochondrial dots, rods and filaments in the ependymoma cells were smaller, finer and fewer. In addition, ependymoma cells in mitosis appeared to have lower succinic dehydrogenase activity and to be poorer in mitochondria than cells in the interphase. 6. It is postulated that the low enzymatic activity observed in ependymoma homogenates is related mainly to the scarcity of mitochondria and in part to the high incidence of mitotic cells.
ENZYMES CATALYZING SEQUENTIAL REACTIONS IN MOUSE BRAIN AND LIVER SUPERNATANT FRACTIONS: I. DIFFERENTIAL USE OF JANUS GREEN B AND PHENAZINE METHOSULFATE
Authors: DU BUY HERMAN G. , SHOWACRE JANE 
Published: April 2011

(1) Janus green B and phenazine methosulfate were used to determine the presence of single enzymes, or enzyme systems, which catalyze sequential reactions in brain and liver supernatants. (2) Spectrophotometric studies on the reduction of Janus green B by supernatants demonstrated the absence of functional flavoprotein in brain supernatants and their presence in liver supernatants, corroborating results obtained by manometry. (3) Brain supernatants contained at least 0.5 microgram of total flavin per ml equivalent; liver supernatant contained about 7 micrograms per ml equivalent. Therefore, the inability of brain supernatant to react with Janus green B was due not to the absence of riboflavin or flavin-adenine dinucleotide, but to the absence of a functional flavoprotein. (4) The absence of a functional flavoprotein did not eliminate the activity of the glycolytic enzyme system in brain supernatants as measured by substrate-dependent lactate accumulation. (5) A flavin inhibitor, acriflavin, did not significantly block glycolysis in brain or liver supernatants, either in the presence or absence of Janus green B. (6) There was no evidence for the presence of a soluble flavin inhibitor in brain supernatants. (7) Reduced diphosphopyridine nucleotide, used as substrate, was oxidized endogenously at a slow rate by both supernatants. This rate was increased by Janus green B in liver but not in brain supernatants. (8) In the presence of pyruvate and Janus green B, both supernatants rapidly oxidized reduced diphosphopyridine nucleotide, but the dye remained oxidized. This indicates that the flavindye-oxygen electron transport chain was inoperative in the presence of the lactic dehydrogenase-pyruvate system of both supernatants. (9) Where the absence of functional flavoprotein prevented the use of Janus green B for the demonstration of enzyme systems, phenazine methosulfate was used. Oxygen uptake in the presence of this dye was a measure of reduced diphosphopyridine nucleotide formed. Thus, it was a measure of the activity of enzymes or enzyme systems, of which diphosphopyridine nucleotide-requiring dehydrogenases are components. (10) By supplying phenazine methosulfate as terminal electron transport agent, both supernatants were shown to contain enzyme systems which metabolized fructose diphosphate, lactate, and malate. (11) The results are discussed with reference to the thesis that the enzyme systems occurring in supernatants represent fractions of those retained by the corresponding mitochondria.
OBSERVATIONS ON THE DIAZOTIZATION-COUPLING REACTION FOR THE HISTOCHEMICAL DEMONSTRATION OF TYROSINE: METAL CHELATION AND FORMAZAN VARIANTS
Authors: GLENNER G. G. , LILLIE R. D. 
Published: April 2011

1. With the finding of the photosensitivity of protein-bound diazotyrosin, a reproducible and sensitive reaction for tyrosine and certain other phenolic compounds in tissue sections based on diazotization and coupling is presented. 2. The specificity of the diazotization-coupling reaction for protein-bound tyrosine among the amino acids is established and the histochemical localization of tyrosine-containing protein in numerous tissues is described. 3. A modification producing a formazan product is presented, which, however, is too insensitive for exacting histochemical use. A modification producing o,o’-dihydroxyazo compounds, which give excellent histochemical localization and are capable of metal chelation, is described, and the possible use of the uranyl chelate for the demonstration of tyrosine by electron microscopy is suggested.
ELASTOLYTIC EFFECT OF PEPSIN
Authors: FISHER EDWIN R. , ROSENTHAL T. B. , LANSING ALBERT I. 
Published: April 2011

Pepsin exhibits an elastolytic effect on elastic fibers of human skin and aorta in tissue sections fixed in alcohol or formalin which precedes its well recognized collagenolytic action in such preparations. Elastin obtained from ligamentum nuchae is similarly solubilized by pepsin in vitro. Its action has been estimated to be approximately one-eighth that of several elastase preparations tested. The susceptibility of elastic fibers and elastin to peptic digestion suggests that they possess the aromatic acid residues tyrosine and phenylalanine in combination with glutamic acid. Carboxyl groups do not appear essential for this peptic activity since prolonged methylation failed to inhibit the elastolytic effect of pepsin on elastic fibers in tissue sections.
HISTOCHEMICAL DEMONSTRATION OF UNSATURATED LIPIDS BY BROMINE SILVER METHOD
Authors: MUKHERJI MRITYUNJOY , DEB CHANDICHARAN , SEN PARIMAL BIKAS 
Published: April 2011

A method of demonstration of unsaturated lipids in tissue sections has been described. Formol fixed frozen sections are exposed to bromine vapour. It is then allowed to react with an alcoholic or ammoniacal solution of silver nitrate, the silver salt being finally reduced to metallic state by reducing agent. Evidences have been put forward to prove the superiority of bromine over iodine treatment. If properly controlled, the method appears to be quite specific.
ESTERASE ACTIVITY IN LEUKOCYTES DEMONSTRATED BY THE USE OF NAPHTHOL AS-D CHLOROACETATE SUBSTRATE
Authors: MOLONEY WILLIAM C. , MCPHERSON KENNETH , FLIEGELMAN LILA 
Published: April 2011

1. Human myeloid cells in the peripheral blood and bone marrow contain a very strong esterase activity which can be demonstrated histochemically by the use of naphthol AS-D chloroacetate as substrate. 2. As first pointed out by Gomori, this esterase activity is found not only in segmented but also immature myeloid cells. However, stem cells, cosinophilic and basophilic granulocytes, lymphocytes, monocytes, plasma cells, megakaryocytes, adult red cells and red cell precursors do not stain with this method. 3. Histocytes and mast cells are strongly positive for this esterase activity. 4. No differences in esterase activity have been noted thus far between neutrophilic leukocytes in normal individuals and in patients with chronic myclogenous leukemia or other diseases. 5. The leukocyte esterase activity is abolished by various well known inhibiting agents. Activity in young myeloid cells in non-leukemic bone marrow smears and in the peripheral blood of patients with myeloid leukemia is inhibited less than that in adult segmented neutrophils. 6. The functions of this esterase in leukocytes are unknown, but further studies of this enzyme seem warranted.
OSMIUM TETROXIDE AND THE MARCHI METHOD: REACTIONS WITH POLAR AND NON-POLAR LIPIDS, PROTEIN AND POLYSACCHARIDE
Authors: ADAMS C. W. M. 
Published: April 2011

1. The reactions of tissues with OsO4 and the Swank-Davenport KClO3-OsO4 (Marchi) reagent have been studied by blockade of the reducing groups of unsaturated lipid, protein and polysaccharide. Only blockade of the —CH=CH— bond prevented osmiophilia after brief staining (18 hr.). 2. Tissue-protein is neither stained nor demonstrably impregnated after brief treatment with OsO4. 3. Chromatographic separation of degenerating myelin revealed that the Marchi-substance is eluted in the esterified cholesterol fraction. 4. Unsaturated lipid but not protein or polysaccharide are stained by brief treatment with OsO4. The Marchi reaction distinguishes hydrophilic from hydrophobic unsaturated lipid.
HISTOLOGY AND CYTOCHEMISTRY OF HUMAN SKIN. XX. THE DISTRIBUTION OF MONOAMINE OXIDASE
Authors: YASUDA KENJIRO , MONTAGNA WILLIAM 
Published: April 2011

1. Using the method of Glenner et al. (8), we have demonstrate abundant monoamine oxidase in human skin. 2. In the epidermis, the reaction occurs in the cells of the malpighian layer. In all hair follicles, regardless of their states of activity, the wall of the pilosebaceous canal is intensely reactive. In growing follicles, the cells of the outer root sheath contain moderate enzyme activity particularly concentrated in the cells at the periphery, in the region above the bulb; small reactive granules are sparsely distributed in the cells of the dermal papilla. In quiescent follicles, there is moderate activity in the cells of the epidermal sac; the hair germ and the dermal papilla have a scant reaction. In the sebaceous glands, the duct and the undifferentiated cells at the periphery of the acini have a very strong reaction. Both apocrine and eccrine glands show strong enzyme activity. 3. Many cutaneous nerves contain monoamine oxidase. Reactive nerves form plexuses just beneath the epidermis, at the level of the sebaceous gland, and midway along the depth of the hair follicle; reactive nerves are also found around both eccrine and apocrine glands. Eccrine glands are surrounded by a richer bed of nerves than the apocrine glands are. The nerves to the arteries also contain the enzyme. 4. The localization of monoamine oxidase in human skin resembles that of succinic dehydrogenase; the cutaneous nerve fibers and the myoepithelial cells, which contain no succinic dehydrogenase, do, however, contain monoamine oxidase. 5. Many cutaneous nerve fibers, sensory and motor, seem to be reactive for both monoamine oxidase and cholinesterase, We do not know what this means in terms of physiology.
THE DISTRIBUTION OF CHOLINESTERASE IN THE CUTANEOUS RECEPTOR ORGANS, ESPECIALLY TOUCH CORPUSCLES OF THE HUMAN FINGER
Authors: CAUNA NIKOLAJS 
Published: April 2011

1. Cutaneous receptor organs of the human digital skin have been investigated for cholinesterase activity by a modification of the histochemical technique of Koelle and Friedenwald, and the findings have been analyzed in relation to the structure of the receptors. 2. It was found that in Meissner's corpuscles the nerves and their sheaths were negative, but the plasma membranes of the laminar cells and the intercellular substance gave strong positive reactions with either acetyl or butyryl substrates. 3. In Merkel's corpuscles, nerve endings and the associated Merkel's cells were negative, but the preterminal medullated nerve fibres gave positive reactions with the butyryl substrate. 4. In hairs, the terminal nerve fibres surrounding the epithelial sheaths gave positive reactions with both substrates. 5. Free nerve endings were negative but the cells related to the endings gave a positive reaction with both substrates. 6. Capillaries and sebaceous glands were negative, but adventitia of larger blood vessels gave positive reactions with the butyryl substrate. 7. The significance of the findings is discussed and a suggestion is made that both acetylcholine and its enzyme may constitute active agents in the synaptic transmission of a nervous impulse in Meissner's corpuscle.
CORRELATION OF HISTOCHEMICAL, AUTORADIOGRAPHIC, AND MICRORADIOGRAPHIC DEMONSTRATIONS OF TISSUE CALCIFICATION
Authors: JOHNSTON MURIEL E. , DURBIN PATRICIA W. , ASLING C. WILLET 
Published: April 2011

1. The sensitivity and reliability of three representative histochemical tests for calcium (von Kóssa, ferrocyanide, and alizarin) have been compared by applying them to vertebral ossification centers in the rat fetus. 2. The histochemical results were controlled by concurrent study of calcium45 autoradiographs and of soft x-ray microradiographs of the same or adjacent sections. 3. Microradiographs and von Kóssa-positive reactions produced essentially identical representations of mineralized bone and cartilage. 4. The von Kóssa reaction coincided with high grain density regions of calcium45 deposition in stripping-film autoradiographs but did not reflect the sparser densities. 5. The ferrocyanide reaction showed gradients of staining density corresponding in many sites with the varying degrees of radiocalcium density. Faint ferrocyanide reactions extended beyond the limits of microradiographic mineral opacity. 6. The resolution obtained with contact autoradiographs and alizarin dye-lake histological preparations was inadequate for study at high magnification.
AN AUTORADIOGRAPHIC STUDY OF THE DISTRIBUTION OF ZINC-65 IN RAT TISSUES
Authors: MILLAR M. J. , VINCENT N. R. , MAWSON C. A. 
Published: April 2011

A new method of preparing rats for 65Zn autoradiography has been described. By ensuring a low excretion rate of 65Zn, autoradiographs with good cellular detail were obtained. The method used for mounting stripping film prevented contact of water with the tissue except during floating-out of the sections. Autoradiographs of the lateral lobes of the dorsolateral prostate, epididymis, testis, ileum, pancreas and coat hair follicles are illustrated. 65Zn was concentrated at the base and lumen surfaces of the acinar cells in the lateral lobes of the dorsolateral prostate, in the tail pieces of testicular spermatozoa, in epididymal spermatozoa, in the Paneth cells of the ileum, in the pancreatic islets of Langerhans and in the shaft of the hair follicles. 65Zn distribution in liver, submaxillary glands, esophagus, pituitary, adrenals and kidney is also described.
A CYTOCHEMICAL ANALYSIS OF THE LACTIC DEHYDROGENASE-DIPHOSPHOPYRIDINE NUCLEOTIDE-DIAPHORASE SYSTEM IN THE EPIDIDYMIS OF THE MOUSE
Authors: ALLEN JOHN M. , SLATER JUDITH J. 
Published: April 2011

The properties of diphosphopyridine nucleotide diaphorase and the lactic dehydrogenase-diphosphopyridine nucleotide diaphorase system in the epididymis of the mouse have been investigated by cytochemical and quantitative methods. In normal mice diphosphopyridine nucleotide diaphorase activity was homogeneously distributed within the cytoplasm of the epithelial cells of the epididymal canal. A gradient of activity existed along the duct, with the lowest levels of activity appearing in the head segments and the highest levels in the body and tail segments. The lactic dehydrogenase-diphosphopyridine nucleotide diaphorase system differed in its cytological distribution from diphosphopyridine nucleotide diaphorase. In the cells of the body portion of the epididymal canal this system showed high levels of activity in the cell apex with low levels of activity prevailing in the general cytoplasm. In other areas the system showed low levels of activity homogeneously distributed throughout the cytoplasm. Cytochemical studies indicated that the activity of diphosphopyridine nucleotide diaphorase was depressed by orchidectomy and restored by testosterone propionate administration. The activity of this enzyme was unaffected by section of the vasa efferentia (vasectomy). The activity of the lactic dehydrogenase-diphosphopyridine nucleotide diaphorase system was depressed by vasectomy and orchidectomy. Testosterone propionate restored activity of tissues from castrated animals to levels prevailing in vasectomized animals but did not re-establish the distribution pattern observed in normal animals. Quantitative determinations confirmed and extended cytochemical observations.
ACROLEIN-SCHIFF, A NEW STAINING METHOD FOR PROTEINS
Authors: VAN DUIJN P. 
Published: April 2011

A cytochemical method based on the reaction of acrolein with tissue compounds followed by staining with the Schiff reagent is described. The reaction is completely blocked by acetylation. Filter paper spot tests of carbohydrates, nucleic acids and proteins revealed a high specificity for proteins. All proteins except the argininerich protamines were positive in the concentrations tested. Carbohydrates, as well as ribonucleic acid, were negative; deoxyribonucleic acid reacted weakly, probably as a result of a contamination. Tests with compounds of low molecular weight revealed that amino acids were positive, nucleic acid derivatives negative. Arguments are given that the double bond of acrolein reacts with SH, with aliphatic NH2 and NH groups, and with the imidazole group. The use of this method as a cytochemical stain in combination with Feulgen, thionine-SO2 staining is indicated, especially for chromosome studies. The method also seems applicable to paper and agar electrophoresis.
THE SECTION FREEZE-SUBSTITUTION TECHNIQUE: I. METHOD
Authors: CHANG JEFFREY P. , HORI SAMUEL H. 
Published: April 2011

A new technique of freeze-substitution, called "Section Freeze-Substitution," is described. Thin pieces of tissue are quick-frozen as for freeze-drying, and sectioned in a cryostat. The free sections are kept in dry ice-temperature absolute acetone overnight, then picked up with cover glasses, coated with celloidin if necessary, and air dried. The sections adherent to the cover glasses can be stained or incubated in substrate solution directly at room temperature (23-27°C). Finished slides can be made front fresh tissues [See figure in the PDF file] within 24 hours. With few exceptions, both oxidative and hydrolytic enzymes, soluble isotopes, and other chemical substances can be precisely demonstrated in a single substitution process. The cytology of the frozen-substituted tissue is excellent. No other histochemical tissue processing method preserves so much of a particular enzyme. The procedure is simple and the results are highly reproducible.
ROLE OF ESTERATIC INHIBITION ON LOCALIZATION OF ESTERASE AND THE SIMULTANEOUS CYTOCHEMICAL DEMONSTRATION OF INHIBITOR SENSITIVE AND RESISTANT ENZYME SPECIES
Authors: SHNITKA THEODOR K. , SELIGMAN ARNOLD M. 
Published: April 2011

Evidence is presented to show that the selective demonstration of perinuclear and peribiliary cytoplasmic droplets in rat kidney and liver by Holt with halogen-substituted indoxyl acetates is largely due to inhibition of a sensitive esterase in the cytoplasm in excess of the inhibition of a resistant esterase in the droplets. The inhibition comes from the incorporation of ferro-ferricyanide in the substrate medium to serve as an oxidation catalyst for the capture reaction. Analogous results were obtained by using other substrates such as α-naphthyl acetate and naphthol-AS acetate and other inhibitors such as formalin fixation, di-isopropyl fluorophosphate, NaF and arsanilate. A product of this study was the development of a method for the simultaneous cytochemical demonstration of inhibitor-sensitive and inhibitor-resistant species of esterase, by taking advantage of red and blue azo dyes that can be formed from naphthol-AS. The distribution of these two species of esterase is described in some detail in rat kidney and liver and the findings are discussed in relation to biochemical data from cell fractionation studies.
OBSERVATIONS CONCERNING THE PRESENCE OF ISOCITRIC DEHYDROGENASE IN MITOCHONDRIA
Authors: SHEPHERD JAMES A. 
Published: April 2011

The intracellular distribution of rat and human liver isocitric dehydrogenase is essentially the same as that of mouse and rabbit liver. This enzyme is largely cytoplasmic but washing does not remove all of the activity from the mitochondria. A method is outlined using permeability considerations as a test of whether an enzyme is within the mitochondria or adsorbed thereon. This method, when applied to washed liver mitochondria, indicates that some isocitric dehydrogenase activity is within the liver mitochondria of rat and rabbit. The results were not conclusive with human liver.