bluuebunny/crossref_metadata_2025_split huggingface dataset:
HISTOCHEMICAL STUDIES OF SUCCINIC DEHYDROGENASE OF THE BRAIN OF MICE, RATS, GUINEA PIGS AND RABBITS
Authors: SHIMIZU NOBUO , MORIKAWA NAGAAKI 
Published: April 2011

Succinic dehydrogenase activity of the brain was histochemically investigated in mice, rats, guinea pigs and rabbits. Fresh frozen sections cut at 20 µ were stained with the Seligman-Rutenburg method as modified by Rosa and Velardo. The pattern of enzyme distribution is remarkably similar but the intensity of staining differs somewhat in the animal species, being highest in mice and rats, intermediate in rabbits and lowest in guinea pigs. The enzyme activity is found exclusively in the grey matter and ocurs both in the perikaryon and neuropil. The activity in the grey matter exhibits the characteristic local variations which are ascribed to the varying participation of the nerve cell bodies and surrounding neuropils in the reaction. Nerve cell bodies contain variable amounts of dye particles of different sizes, and the neuropil possesses variable numbers of coarse granules. An intense reaction is given in the caudate nucleus, putamen, anterior nucleus of the thalamus, interpeduncular nucleus and cerebellar cortex. An intermediate activity occurs in the neocortex, rhinencephalon, optic tectum, most of the cranial nerve nuclei, superior and inferior olivary nuclei, and the Goll and Burdach nuclei. A low activity is present in the paraventricular structures, such as supraoptic crest, subfornical and subcommissural bodies, and area postrema, as well as the paraventricular and supraoptic nuclei of the hypothalamus, and dorsal nuclei of the vagus nerve.
STUDIES ON TISSUE FIXATION: THE PENETRATION OF TRICHLORACETIC ACID SOLUTIONS INTO RAT TISSUES
Authors: CAFRUNY E. J. 
Published: April 2011

A method for measuring the rate of penetration of trichloracetic acid solutions into tissues was developed and the effects of a surface active agent, temperature, and concentration of fixative on penetration rates were evaluated. Penetration distances into rat liver, kidney, and cerebral cortex were determined and expressed as a function of time. It was found the penetration distances vary directly with temperature and fixative concentration. Tergitol, a wetting agent, did not increase penetration rates into kidney or liver tissue and in high concentration actually decreased penetration into kidney. Penetration distances were found to be proportional to the square root of the time of exposure of liver to 10% trichloracetic acid solutions but this proportionality did not hold for kidney and cerebral cortex.
ULTRACENTRIFUGAL STUDIES ON THE CYTOPLASMIC COMPONENTS
Authors: TAKANAMI MITURU 
Published: April 2011

Ultracentrifugal analysis, electron microscopical observation and chemical analysis on the cytoplasmic homogenate of rabbit liver were performed in order to determine the constitution and diversity of the cytoplasmic components. The result of experiments showed that the cytoplasmic components comprised 1) a heterogeneous granular component, sedimented at a rate more than 250s, 2) two closely spaced components whose sedimentation rates were 51s and 64s respectively and 3) three smaller components with rates of 4s, 6s and 12s with regard to the sedimentation analysis. The first heterogeneous granular component showed continuous distribution of size and shape from 50-60 mµ to several µ in diameter and contained three chemical materials, ribonucleic acid, lipids and protein components. The next two closely spaced components, corresponding with considerable numbers of homogeneous micro-particles of about 20 mµ electron microscopically, represented a ribonucleoprotein component in which the RNA:protein ratio was about 1:1.7 and a metalloprotein containing about 20% iron respectively. Accordingly, I must emphasize the need to discriminate these macromolecular components from the heterogenous granular component, including the so-called large granule and small granule fractions, for biochemical Studies on the cell components, and that the cell-fractionation techniques should be improved according to present ultracentrifugal data.
INTRACELLULAR LOCALISATION OF DEHYDROGENASE SYSTEMS USING MONOTETRAZOLIUM SALTS AND METAL CHELATION OF THEIR FORMAZANS
Authors: PEARSE A. G. E. 
Published: April 2011

1) A method is proposed for the demonstrations of dehydrogenases and diaphorases using the principle of simultaneous chelation of an enzymically produced monoformazan by means of metallic ions. 2) Using 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyl tetrazolium and M/60 cobalt a black metal-formazan complex is produced. Its nature is such that fine intracellular localisation becomes possible. 3) Chelation occurs under histochemical conditions with cobalt, copper, nickel and uranium and it is suggested that the electron opacity so induced may facilitate localisations of dehydrogenases on the electron microscope level.
TETRAZOLIUM REDUCTION AS A MEASURE OF MONOAMINE OXIDASE ACTIVITY IN VITRO
Authors: WEISSBACH HERBERT , REDFIELD BETTY G. , GLENNER GEORGE G. , MITOMA CHOZO 
Published: April 2011

1 . Data has been presented showing that tetrazolium salts can be used for the histochemical localization of monoamine oxidase, although there are limitations to its application to in vitro studies. These limitations have been discussed. 2. It appears that an indole carbonyl compound is necessary for 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyl tetrazolium chloride reduction. A diaphorasetype system may be required to transfer the electrons from the indole carbonyl intermediate to INT.
REACTION OF TISSUES WITH SCHIFF'S REAGENT AFTER TREATMENT WITH ANHYDROUS CHROMYL CHLORIDE
Authors: MENDE THOMAS J. , CHAMBERS EDWARD L. 
Published: April 2011

Oxidation with chromyl chloride in an anhydrous solvent followed by exposure to Schiff's reagent can be used for the demonstration of glycogen and mucins in tissue sections. The intensity of the reaction is weaker than in the periodic acid-Schiff technique and other substances besides glycogen and mucins generally fail to react. Acetylation and deacetylation abolish and restore the reaction respectively. On prolonged exposure to chromyl chloride at room temperature a typical Feulgen nucleal reaction is obtained. Treatment with the following inorganic acid chlorides in carbon tetrachloride also results in Feulgen staining: sulfuryl chloride thionyl chloride and phosphoryl chloride. Current views concerning the mechanism of aldehyde group liberation in the Feulgen reaction are discussed in connection with these results.
IMPROVEMENT IN THE HISTOCHEMICAL LOCALIZATION OF SUCCINIC DEHYDROGENASE BY USE OF NITRONEOTETRAZOLIUM CHLORIDE
Authors: PEARSON BJARNE 
Published: April 2011

A new method for the histochemical localization of succinic dehydrogenase activity has been described in which the resultant formazan is free of crystal formation. The resultant formazan is extremely fine and granular permitting accurate and precise localization of the enzyme. The tetrazolium salt used was 2,2'-diphenyl-5,5' - di - ( m - nitrophenyl) - 3,3' - (4,4' - biphenylene) ditetrazolium chloride. Minimal granular artifacts which appear may be eliminated by dissolving the tetrazolium salt in N,N-dimethylformamide and rinsing the slide in 80% alcohol following incubation. This has a tendency to dissolve fat and prevent aggregation of granular artifacts which may appear subsequently upon storage of slides.
THE NILE BLUE REACTION OF PEPTIC GLAND ZYMOGEN GRANULES: THE EFFECT OF METHYLATION AND ALKALI DEMETHYLATION
Authors: LILLIE R. D. 
Published: April 2011

The zymogen granules of the chief cells of the peptic glands are colored dark blue when the sulfuric acid Nile blue technic for the demonstration of lipofuscin is applied. Weaker blue colors appear in smooth muscle, soft keratin and certain cytoplasms. After methylation the same technic colors these structures in varying shades of pink to red. Decomposition of the methyl esters by KOH restores the blue staining. Extraction with acetone directly after staining with sulfuric acid Nile blue prevents both the blue and the pink to red colorations. The strong Nile blue staining of peptic gland zymogen granules may correlate with the large excess of free anionic residues over cationic in pepsin, and the considerable excess of aspartic and glutamic acids over arginine, histidine and lysines in the same protein. Use of alcoholic potassium hydroxide affords a convenient histochemical procedure for conversion of esters to fatty acids, provided the acids are not soluble in the alkali.
A NEW CARBOWAX METHOD FOR ROUTINELY PERFORMING LIPID, HEMATOXYLIN AND EOSIN AND ELASTIC STAINING TECHNIQUES ON ADJACENT FREEZE-DRIED OR FORMALIN-FIXED SECTIONS
Authors: ZUGIBE FREDERICK T. , KOPACZYK KRYSTYNA C. , CAPE WILLIAM E. , LAST JULES H. 
Published: April 2011

1. A freeze-drying method is presented for the simultaneous processing of a large number of tissue specimens. 2. An infiltration medium consisting of carbowax "1000" and a blocking mixture consisting of carbowax "1000" and carbowax "4000" were developed and resulted in ribbons which could be cut as fine as 2 µ. 3. Diffusion currents encountered during flotation were obviated by the use of a General Flotation Solution consisting of potassium dichromate, glycerin and gelatin for routine sections and a Special Flotation Solution consisting of carbowax and glycerin for certain fragile tissues. 4. The application of routine stains to adjacent carbowax sections included a hematoxylin and cosin technique with good cosin differentiation and sharp hematoxylin staining, an elastic-collagen technique demonstrating distinct elastic and collagen fibers and a lipid technique with minimal fat loss. 5. This carbowax method allows for correlative, multiple staining and ultramicrochemical analysis on adjacent tissue sections.
A SEMI-MICRO METHOD FOR THE DETERMINATION OF CRYSTAL VIOLET SORBED TO BIOLOGICAL MATERIAL
Authors: FISCHER ROLAND , ZALESCHUK JERRY 
Published: April 2011

A semi-micro method is reported to measure the degree of Gram positive behaviour, i.e. the amount of crystal violet sorbed by various proteinous materials. Part of the role supposedly played by "loosely bound" iodine as well as that by covalently bound iodine in the crystal violet uptake of intact wool (Gram negative) and alkali-degraded wool (Gram positive) has been determined. The influence of reversing the consecutive steps of staining and iodination on dye-uptake has been measured for wools. The iodine uptake prior to and after staining, as well as the dye-uptake prior to iodination, have been determined for the two types of wool as well as for Escherichia coli and Leuconostoc mesenteroides respectively. The dye-uptake of wools and of bacteria was also measured after treatment with 1-fluor-2,4-dinitrobenzene. The identical composition in non-sulphur containing amino acids of intact wool and alkalidegraded wool—prior to and after treatment with 1-fluor-2,4-dinitrobenzene—has been investigated by subjecting the wools to hydrolysis and one- as well as two-dimensional paper chromatography. Data are reported as to the crystal violet uptake prior to and after iodination of muscle and nerve-fibres, nail, ovalbumin, β-glucuronidase and pepsin. The theoretical implications of these and other data are discussed and attention is drawn to the manyfold aspects of the problem of Gram staining behaviour of proteins.
CHARACTERIZATION OF SCHIFF-POSITIVE SUBSTANCES IN ELASTIC FIBERS
Authors: LABELLA FRANK S. 
Published: April 2011

Schiff-positive substances in clastic fibers have been separated on the basis of differences in reactivity with the reagent. Alkali-treated ligamentum nuchae powder contains a cerebroside which is removed by chloroform or pyridine. Also present are a plasmalogen-like compound which depends upon its unsaturated component for its reactivity, and a substance whose Schiff positivity is negated by prior treatment with iodine; both reactants are resistant to extraction techniques. Mercuric chloride or enzymatic hydrolysis releases an unsaturated fatty material, probably the aldehydic component of the clastin-bound plasmalogen-like compound. Elastin powder which has been treated with phenylhydrazine, iodine and bromine still gives a positive reaction with fuchsin-sulfurous acid. Rat aortic clastica, like ligamentum nuchac, contains a non-extractable plasmalogen like compound whose reactivity with Schiff's reagent depends Upon unsaturation in the molecule. Staining intensity is reduced by prior chloroform extraction, suggesting a glycolipid as was demonstrated for ligamentum nuchae.
ON THE EXPERIMENTAL DISCHARGE OF THE GASTRIC AND INTESTINAL ARGENTAFFIN CELLS
Authors: LILLIE R. D. , WEISSBACH H. , GLENNER G. G. 
Published: April 2011

Reserpine discharges the azo positive reducing substance from both gastric and intestinal basal granular cells in the guinea pig. Regeneration was slower than in previous reports and apparently more rapid in the duodenum than in the stomach or ileum. The numbers of cells demonstrated in the normal duodenum and ileum respectively, as well as in the duodenum and ileum of the 7 day post reserpine animal, do not correspond particularly closely with the 5-hydroxytryptamine assays. Physostigmine and ephedrine failed to discharge the reactive material from basal granular cells of the guinea pig stomach and intestine. Argyrophil cells persist throughout the reserpine discharge period, during which argentaffin, azo positive basal granular cells are not demonstrable.
THE DISTRIBUTION OF ESTERASES IN MOUSE TISSUES
Authors: MARKERT CLEMENT L. , HUNTER ROBERT L. 
Published: April 2011

The esterases contained in 32 mouse tissues and organs have been analyzed using the zymogram technique. More than ten different esterases have been resolved and partly characterized by their substrate and inhibition specificities. The esterases compose a family of distinct enzymes with overlapping specificities, and the total activity in any tissue is the sum of the activities of the particular esterases in that tissue. Characteristic similarities and differences among the esterase patterns from different tissues were noted and discussed. The G band is apparently a cholinesterase as shown by its sensitivity to inhibition by eserine at 10–4 M. The small effect of age, sex, genotype, and nutrition upon the esterase pattern is contrasted with the striking changes occurring during the differentiation of organs. The greatest change not associated with differentiation was observed in blood after 5 days of starvation. Fractionation of liver homogenates by centrifugation showed esterase to be concentrated primarily in the microsome fraction. Measurements were made of the quantitative distribution of esterase activity among the electrophoretically resolved bands from each of the centrifugal fractions of liver homogenates. The pattern of esterases was very similar in each of the centrifugal fractions.
NEW HISTOCHEMICAL TECHNIQUES FOR THE DEMONSTRATION OF TISSUE OXIDASE (CYTOCHROME OXIDASE)
Authors: BURSTONE M. S. 
Published: April 2011

A group of complex naphthols, as well as open chain and cyclic methylene compounds, were found to be useful new reagents for the demonstration of tissue oxidase in conjunction with N-phenyl-p-phenylenediamine. The methylene compounds constitute a new class of histochemical reagents. N-alkyl amines (e.g. N,N-dimethyl arylamines) are toxic to the enzyme and are to be avoided. Complex amines are inhibitory and are likewise not recommended. As with the original "Nadi" technique, the reaction was inhibited by cyanide, azide, and sulfide. The distribution of staining in tissues was essentially identical with all reagents. Although the reaction was augmented by cytochrome c, this expensive reagent is not neeeded with the present techniques. The indoaniline and azamethine dyes produced are highly insoluble and result in permanent or relatively permanent preparations. Most of the dyes are capable of complexing with mercury, molybdenum, cobalt, nickel, cadmium, lead, uranium, and iron, and thus may have potential applications in electron microscopy. Among the naphthols 1-hydroxy-2-naphthoic acid (IV), 1-hydroxy-2-acetonaphthone (V), and a derivative of 1-hydroxy-2-naphthanilide (IX) are very useful couplers. Useful methylene compounds are exemplified by 1-phenyl-3(m-nitrobenzamidopyrazolone) (XII), naphthol AS-LG (XVIII), and naphthol AS-L3G (XX). Use of methylene compounds results in highly stable incubating solutions. All substrates are commercially available and inexpensive. Unfixed frozen sections mounded on glass slides were used. Considerable caution must be exercised in attempts to localize oxidative enzyme activity at the cytologiocal level in unfixed frozen sections. This is due to the disruption of mitochondria and other cytological constituents which may occur during freezing and thawing. An alternative technique was evolved. This consisted of incubating fresh tissue slices, freezedrying them, followed by embedding in paraffin. Sections were then cut, dewaxed, and mounted. Preliminary results indicate that tissue oxidase may also be demonstrated using a mixture of amines (e.g., N-phenyl-p-phenylenediamine and 2-methoxy-N-phenylenediamine) which are capable of forming complex azine dyes. The possibility of demonstrating other enzymes of the oxidase dehydrogenase complex warrants further investigation.
STUDIES OF SULPHUR, PHOSPHORUS AND SOME METALS IN SKIN FOLLICLES
Authors: RYDER M. L. 
Published: April 2011

Radioactive sulphur could be detected rat and mouse follicles sooner after injection than in the lamb, and this may be associated with the faster growth rate of hair in the rat and mouse. It was not possible to detect copper in follicles, although zinc was detected and found to be concentrated in the unkeratinised part of the fibre. Potassium was found in the outer sheath and in the unkeratinised part of the fibre, as well as in the stratum corneum. Labelled phosphorus was detected mainly in the bulb, with smaller amounts in the unkeratinised part of the fibre.
THE DISTRIBUTION OF DEOXYRIBONUCLEASE IN NORMAL, CIRRHOTIC AND NEOPLASTIC RAT LIVERS
Authors: DAOUST R. , CANTERO A. 
Published: April 2011

The distribution of deoxyribonuclease in normal, cirrhotic and neoplastic rat livers was investigated histochemically using the gelatine-DNA film method. The bile duct cells and connective tissue elements which are present in abnormal amounts in cirrhotic liver contain little DNAase activity compared with parenchymal tissue. The distribution pattern of DNAase is relatively uniform in normal liver parenchyma but becomes heterogeneous in the parenchyma of cirrhotic and neoplastic tissues. Groups of parenchymal cells in cirrhotic liver and the hepatoma cells in general appear devoid of DNAase activity. The necrotic areas of tissues, on the other hand, show intense enzyme activity.
ALDEHYDE-FUCHSIN FOR PITUITARY CYTOCHEMISTRY
Authors: ELFTMAN HERBERT 
Published: April 2011

The component of the aldehyde-fuchsin stain which is useful for pituitary staining requires more delicate manipulation than the elastic tissue component. Manufacture of the stain for pituitary purposes may he shortened to 26 hours by subjecting a 0.5% basic fuchsin solution in 70% ethanol, 0.75% paraldehyde and 1.25% HCl to incubation at 37°C. The pH of this solution precludes background staining, allowing aldehyde-fuchsin to be followed by the periodic acid-Schiff routine. Aldehyde fuchsin may be rejuvenated by adding basic fuchsin in alcoholic solution. Oxidation of pituitary tissue allows aldehyde-fuchsin to stain not only thyrotrophs but gonadotrophs as well and accentuates the staining of the hypothalamic hormones and the Golgi bodies. Since some fixatives, such as dichromates, oxidize tissue, assay of the significance of aldehyde-fuchsin staining involves awareness of the influence of the particular fixative employed. Spectrophotometric analysis of young and old aldehyde-fuchsin stains supports the assumption that staining of the pituitary thyrotrophs may involve other constituents of the stain complex than those relied on for elastic tissue visualization.
SPECIFICITY OF LABELLED THYMIDINE AS A DEOXYRIBONUCLEIC ACID PRECURSOR IN RADIOAUTOGRAPHY
Authors: AMANO M. , MESSIER B. , LEBLOND C. P. 
Published: April 2011

A simple method has been devised for the extraction of deoxyribonucleic acid from histological sections by means of the enzyme DNAse without significant loss of ribonucleic acid. Removal of deoxyribonucleic acid by this method completely eliminated the radioautographic reactions observed over nuclei after injection of thymidine-H3. It is concluded that labelled thymidine is an adequate tracer of newly formed deoxyribonucleic acid in radioautographs.
LIMITATIONS IN THE HISTOCHEMICAL DEMONSTRATION OF PYRIDINE NUCLEOTIDE-LINKED DEHYDROGENASES ("NOTHING DEHYDROGENASE")
Authors: ZIMMERMANN H. , PEARSE A. G. E. 
Published: April 2011

1. The ability of SH groups in well-dialyzed protein fractions to reduce exogenous pyridine nucleotides has been described in the biochemical literature. 2. This effect has been shown to be the cause of the positive histochemical result obtained when fresh tissue sections are incubated at alkaline pH levels with oxidized pyridine nucleotides (DPN or TPN) and tetrazolium salts, in the absence of any substrate. 3. The effect is pH-dependent, commencing at about pH 7.0 and rising to a maximum between pH 8.0 and 9.0. It is abolished by low concentrations of sulphydryl inhibitors or by heating the tissues. 4. The specific diaphorases are unaffected by sulphydryl inhibitors and these are presumed to transfer hydrogen and electrons from the reduced pyridine nucleotides (DPNH, TPNH) to the tetrazolium salt. 5. The development of histochemical methods for specific pyridine nucleotide-linked dehydrogenases is hindered by the activity described, to which Racker's convenient term "nothing dehydrogenase" has been applied.