bluuebunny/crossref_metadata_2025_split huggingface dataset:
THE SPECIFICITY OF THE HISTOCHEMICAL METHOD FOR ADENOSINE TRIPHOSPHATAS
Authors: PADYKULA HELEN A. , HERMAN EDITH 
Published: April 2011

Three categories of phosphatase activity toward adenosine triphosphate at pH 9.4 have been isolated histochemically: (1) phosphatase which requires -SH groups; (2) phosphatase which is inhibited by -SH groups; and (3) phosphatase which is relatively indifferent to -SH groups. This enzymatic activity was demonstrated in thin (5 µ), unfixed, frozen sections. True adenosine triphosphatase has been separated histochemically by virtue of its -SH dependence. It was demonstrated in cardiac and skeletal muscle fibers and in the mitochondria of the kidney tubules. The staining of these structures can be prevented by treatment with an -SH inhibitor, such as p-chloromercuribenzoate or salyrganic acid. Also their stainability can be restored by reversing the inhibition of the enzymes with -SH compounds, such as BAL or cysteine. There was no reaction of these particular structures when adenosine diphosphate or adenosine-5-phosphate was used, suggesting that under the conditions of this histochemical test only the terminal phosphate of adenosine triphosphate is removed. Phosphatase activity of the smooth muscle of the intestinal muscularis toward adenosine triphosphate differed from that of striated muscle in that it was less sensitive to p-chloromercuribenzoate. Sulfhydryl compounds, such as BAL or cysteine, enhanced the staining of the above mentioned sites of true adenosine triphosphatase activity. On the other hand, these -SH compounds are powerful inhibitors of alkaline phosphatase activity, especially toward adenosine-5-phosphate. When adenosine triphosphate was the substrate, BAL prevented the staining of the brush borders of the kidney tubules and intestinal epithelium, thus suggesting the participation of alkaline phosphatase in the dephosphorylation of adenosine triphosphate. Furthermore, the staining of these brush borders was unaffected by the -SH inhibitors. The strong phosphatase activity of endothelium and vascular smooth muscle toward adenosine triphosphate was seemingly indifferent to -SH groups, since the staining of these structures was not markedly influenced by -SH inhibitors or compounds. The nature of this staining has not been elucidated. Comparisons of the localization of phosphatase activity toward adenosine triphosphate, -diphosphate or -5-phosphate were made in the ventricle, tongue, kidney, and duodenum. Differences in the localization of phosphatase activity toward adenosine triphosphate and adenosine-5-phosphate were also made in liver, lung, spleen, uterus, and aorta.
SIGNIFICANCE OF COUPLING RATE IN HISTOCHEMICAL AZO DYE METHODS FOR ENZYMES
Authors: DEFENDI V. , PEARSE A. G. E. 
Published: April 2011

1) In order to estimate the significance of the coupling rate in histochemical localization of enzymes a series of naphthols has been tested, using a streaming method with photoelectric recording. The naphthols were those used in histochemical methods (as esters) and the diazonium salt employed (Fast Blue B salt) was chosen as being the one most often used in histochemistry. 2) It was observed that the usual straight line relationship between the bimolecular velocity constant of coupling and pH (Conant and Peterson, 1930) became invalid at certain "critical points." For β-naphthol the "critical point" lay between pH 7.4 and 7.0 and for α-naphthol between pH 5.3 and 5.0. 3) Parallel histochemical studies indicated that the "critical point" and therefore the coupling rate, was likely to be important in the case of the more soluble naphthols, but of less importance with the relatively insoluble ones, in producing accurate histochemical localization of enzyme.
THE VALUE OF SOME TETRAZOLIUM SALTS AS HISTOCHEMICAL REAGENTS FOR SULPHYDRYL GROUPS
Authors: FINDLAY G. H. 
Published: April 2011

A study was made of the reaction between sulphydryl compounds and tetrazolium salts, to estimate the value of the latter in the histochemistry of sulphydryl groups. The chemical reaction was found to be precarious, the relative concentrations of the two reactants making a great difference to its rate and efficiency. Hydrogen-ion concentration, oxygenation of the solutions, the presence of wetting agents and of other reducing compounds influenced the color development in a complicated manner. Mol-equivalents of various sulphydryl-containing compounds reacted differently towards tetrazolium salts. Histochemical quantitation was consequently difficult, apart from the fact that the reaction was relatively inefficient. Nevertheless, the histochemical results were considered to be comparable with those obtained with dihydroxydinaphythyldisulphide ("DDD"), although the overlap was not complete. Chemical and histochemical results were also at variance. For histochemical purposes the method was most suited to demonstrate disulphide and sulphydryl groups together. Control sections were made in which disulphides alone were stained, and in which both sulphydryl and disulphide groups were blocked. For showing sulphydryl groups alone the reaction was least satisfactory.
A STRICTER INTERPRETATION OF THE FERRIC FERRICYANIDE REACTION WITH PARTICULAR REFERENCE TO THE DEMONSTRATION OF PROTEIN-BOUND SULPHYDRYL AND DI-SULPHIDE GROUPS
Authors: ADAMS C. W. M. 
Published: April 2011

(1) Two distinct pigments (Prussian blue and Prussian green) are found in tissues after the ferric ferricyanide reaction; they have different chemical properties. (2) Only strong reduction causes rapid formation of Prussian blue from ferric ferricyanide solutions. Conversely Prussian green may be formed by weak reduction or possibly by absorption of traces of this pigment present in ferric ferricyanide solutions. Brown or yellow pigments in tissues may appear green in the presence of low concentrations of Prussian blue. Thus no strict interpretation of green colours can be made. (3) Sulphydryl groups, phenols, indoles, pyrroles, aromatic amines, uric acid and ascorbic acid reduced ferricyanide to Prussian blue. Certain oils, aliphatic amines, aldehydes and some inorganic reagents caused the production of Prussian green. (4) Up to 5 minutes immersion in the ferric ferricyanide solution is recommended for the demonstration of sulphydryl groups in trichloracetic-alcohol fixed tissues. (5) A technique is given for the demonstration of protein-bound disulphides, involving previous thioglycollate reduction followed by the demonstration of the formed sulphydryls by 1½ minutes immersion in the ferricyanide solution. This technique demonstrates protein containing at least 4 per cent of cystine (equivalent to 1 per cent of sulphur).
STUDIES ON THE CONNECTIVE TISSUE OF THE COCK COMB: I. HISTOCHEMICAL OBSERVATIONS ON THE GROUND SUBSTANCE
Authors: SZIRMAI J. A. 
Published: April 2011

1. The ground substance of the mucoid layer of the cock comb has been found to show a strongly metachromatic reaction with toluidine blue, but the PAS reaction was negative. The fibrils, which do not stain metachromatically, show a significant PAS positivity. The metachromatic ground substance is very easily soluble in water, while the PAS positive substance of the fibrils is more resistant to extraction. The metachromasia of the ground substance disappears below pH 3.0. No metachromasia could be observed on sections previously treated with gelatin solution. In the capon comb the PAS positivity is restricted to the thick collagen bundles and no metachromatic ground substance is present. 2. The uptake of toluidine blue by the comb tissue was determined quantitatively. The values obtained with sections of the cock comb are about 15 µg/mg. wet tissue, decreasing to about 3 µg/mg. after extraction with dist. water. The capon comb bound about 2-3 µg toluidine blue per mg. wet tissue. 3. Hexosamine determinations on slices of the whole cock comb gave dry weight values of about 4%, while the mucoid layer alone yielded 1½ to 3 times higher values. After extraction with dist. water, the cock comb tissue loses about 75% of the total hexosamine. In the capon comb, a hexosamine content of about 0.5% was found; this value was not influenced by previous dist. water extraction. 4. The water content of the defeated cock comb was found to be about 89% of the dry weight; that of the isolated mucoid layer was even higher (90 and 94%). The water content of the capon comb is about 75%.
THE MECHANISM OF THE MARCHI TYPE OF METHODS FOR VISUALIZING DEGENERATING MYELIN. MODEL EXPERIMENTS WITH PURE COMPOUNDS
Authors: WOLMAN M. 
Published: April 2011

The chemical changes occurring in myelin lipids do not explain the reason for the specificity of Marchi's stain for degenerating myelin. Experiments were made with a number of pure compounds smeared on glass slides with the aim of finding the type of compound which might be responsible for the Marchi staining. Among the compounds tested heparin was the only one which fulfilled the requirements. This finding suggests a working hypothesis for further work on the mechanism of the early changes during demyelinization.
HISTOCHEMICAL DEMONSTRATION OF AMINOPEPTIDASE
Authors: BURSTONE M. S. , FOLK J. E. 
Published: April 2011

A histochemical method for demonstration of aminopeptidase in tissue sections using the substrates l-leucyl β-naphthylamide and dl-alanyl β-naphthylamide is presented. The method may also be quantitated on tissue sections. The successful histochemical demonstration of the enzyme with both substrates is dependent upon the use of frozen-dried tissues.
HISTOCHEMICAL LOCALIZATION OF SPECIFIC OXIDATIVE ENZYMES: II. LOCALIZATION OF DIPHOSPHOPYRIDINE NUCLEOTIDE AND TRIPHOSPHOPYRIDINE NUCLEOTIDE DIAPHORASES AND THE SUCCINDEHYDROGENASE SYSTEM IN THE KIDNEY
Authors: STERNBERG WILLIAM H. , FARBER EMMANUEL , DUNLAP CHARLES E. , LOUVIERE CONCETTA M. 
Published: April 2011

The histochemical distribution of the succinic dehydrogenase system, DPN diaphorase and TPN diaphorase has been studied in the rat kidney. The staining pattern for each enzyme shows a highly reproducible and distinctly different zonal distribution in the cortex and medulla and a corresponding regional distribution along the course of individual nephrons. With these methods, virtually all the different segments of the nephron can be sharply delineated and identified. Collecting ducts in the papilla, unstained with the succinate method, are well stained with DPN and TPN diaphorase. TPN diaphorase gives good staining of the thin limbs of Henle which are so difficult to demonstrate by other methods. With the succinate system and DPN diaphorase, the thin limbs of Henle are unstained, but the thick ascending limbs stain heavily. This accounts for the sharp demarcation in staining seems between the outer medullary zone and the papilla. It is interesting that, with the succinate system, the collecting ducts show an abrupt cessation of staining at the same level. Vascular endothelium and glomeruli show delicate staining with both diaphorases but not with succinate. Similarly, smooth muscle and the epithelium of the renal pelvis are stained by the diaphorase methods, but poorly, if at all, with succinate. A distinct inner non-glomerular zone of the cortex is present in the rat and mouse kidney and is composed principally of coiled terminal portions of proximal convoluted tubules. In these rodents, the terminal segment of the proximal convoluted tubule is not straight and no distinct medullary rays are present. The staining of the inner cortical zone is distinctively different by all three methods from the staining of the outer zone which contains the first portion of of the proximal convoluted tubule. This furnishes additional evidence for a functional division of labor in the proximal convoluted tubule. In the papilla, fusiform cells of unknown function are present in parallel formations, oriented at right angles to the direction of the tubules. These are well demonstrated in the rat kidney by TPN diaphorase and are found to be even more abundant in a desert rodent, the kangaroo rat. Should these cells prove to have a contractile function, they might play a role in the regulation of urinary volume. Studies of the chemical anatomy of the kidney using these enzyme methods offers further evidence that the nephron is divisible into more than the conventional number of functional segments.
HISTOCHEMICAL LOCALIZATION OF SPECIFIC OXIDATIVE ENZYMES III. EVALUATION STUDIES OF TETRAZOLIUM STAINING METHODS FOR DIPHOSPHOPYRIDINE NUCLEOTIDE DIAPHORASE, TRIPHOSPHOPYRIDINE NUCLEOTIDE DIAPHORASE AND THE SUCCINDEHYDROGENASE SYSTEM
Authors: FARBER EMMANUEL , STERNBERG WILLIAM H. , DUNLAP CHARLES E. , LOUVIERE CONCETTA M. 
Published: April 2011

Histochemical stains are methods for the visual demonstration of the biochemical properties of organized tissues. They differ from "ordinary" tissue stains only in so far as they lend themselves to chemical interpretation and are reliable only to the extent that they accurately reflect the presence and localization of specific chemical substrates or activities. In this paper we have described the results of a number of experiments designed to test the reliability and usefulness of tetrazolium salts as histochemical indicators of specific oxidative enzyme activities. Control studies dealing with substrate specificity and inhibitors gave further evidence of the specificity of tetrazolium staining methods for the succinic dehydrogenase system. Studies on the possibility of diffusion and nonspecific deposition of reduced blue tetrazolium and neotetrazolium indicated that the blue reduction products (diformazans) of each do not deposit nonspecifically from the medium and once deposited are firmly fixed in the tissues. The red reduction products (monoformazans) may deposit nonspecifically from the medium. Hence their deposition and localization must be interpreted with caution. The red monoformazans of these two salts can readily be converted to blue diformazans in tissue sections by non-enzymatic reducing agents. Blue tetrazolium in the unreduced form is reversibly bound in the cytoplasm of all cells of the rat kidney but not in the nuclei. In the staining process it is probable that tetrazolium, bound in the cytoplasm, is reduced to formazan irrespective of the continued presence of free tetrazolium in the medium. Since no binding of tetrazolium is demonstrable in any of the cell nuclei, the failure of nuclei to stain does not necessarily indicate the absence of enzyme. It was found that two other hydrogen acceptors, methylene blue and pyocyanin, are also bound by the cytoplasm of cells and when either of these compounds is added to tetrazolium staining media, tissue staining is greatly accelerated and enhanced. Relative enzyme activities determined quantitatively in tissue taken from different anatomical regions of the rat kidney correspond fairly well with the intensity of histochemical staining. Tetrazolium methods are now available for demonstrating the succinic dehydrogenase system, diphosphopyridine nucleotide diaphorase and triphosphopyridine nucleotide diaphorase. These methods appear to satisfy most of the criteria which we have adopted for the specificity and reliability of histochemical stains. They yield preparations which are quite satisfactory for histochemical purposes. However, it has not as yet been demonstrated that the formazan deposits accurately indicate the intracellular localization of enzyme activity and hence cytochemical interpretations are open to question.
CARBOHYDRATE AND CARBOHYDRATE METABOLITE UTILIZATION BY ENZYME SYSTEMS OF MOUSE BRAIN AND LIVER MITOCHONDRIA
Authors: DU BUY H. G. , HESSELBACH M. L. 
Published: April 2011

1. Both mouse brain and mouse liver preparations were found to utilize substrates characteristic of the following carbohydrate enzyme systems : the glycogen, glucose, fructose, mannose, fructose-1-phosphate (5), oxidative pentose phosphate (4), and tricarboxylic acid system. 2. Each of these enzyme systems was found on isolated brain mitochondria and a number on isolated liver mitochondria. 3. The implications of these findings in regard to the lability, normal enzyme system complement and functions of mitochondria are discussed.
ATTEMPTS AT HISTOCHEMICAL DEMONSTRATION OF ETHYLENE GROUPS, WITH PARTICULAR REFERENCE TO STAINING PROPERTIES OF HAIR CORTEX
Authors: LANDING BENJAMIN H. , HALL HAZEL E. 
Published: April 2011

(1) The results of comparative staining of hair cortex, posterior pituitary, and several tissue lipids, with leukofuchsin, aldehyde-fuchsin, and toluidine blue, after a variety of pretreatments, are presented. (2) The results confirm those of Lillie and Bangle, that induced Schiff-positivity of hair cortex is not due to oxidation products of cystine, while the metachromatic basophilia induced in hair cortex by appropriate reagents is so explained. However, whether the reactive group responsible for the Schiff-positivity is a part of keratin or of some other molecular entity has not been established. (3) Preliminary results with four additional reagents capable of producing histochemically demonstrable products from ethylenic groups are presented. Selenium dioxide appears to have little promise for this purpose, and osmic acid potassium chlorate appears to oxidize both lipids and cystine. Iodoform peracetic acid has provided evidence of the occurrence of unsaturated entities, perhaps lipids, in hair cortex. Phosphorus pentachloride may have value as a reagent for demonstration of certain lipids by means of induced basophilia, but the mechansim of phosphate radical binding may well be esterification rather than substitution into ethylenic groups. (4) Aldehyde-fuchsin appears to have particular affinity for certain reactive groups, of unknown nature but possibly neither carbonyl nor acid radicals, which are produced in sections by treatment with sulfuric acid permanganate. (5) Phosphorus pentachloride, like phosphoryl chloride, leads to leukofuchsin-positivity of certain tissue entities. No evidence that this reaction is not due to carbonyl groups was obtained, but phosphorylating agents are of interest with regard to the chemical nature of reactive groups responsible for certain Schiff reactions.
STABILITY OF THE FEULGEN-DEOXYRIBONUCLEIC ACID ABSORPTION CURVE IN SITU WITH VARIATION IN NUCLEAR PROTEIN CONTENT AND OTHER FACTORS
Authors: KASTEN FREDERICK H. 
Published: April 2011

1. The influence of certain variables upon the shape of the Feulgen absorption curve of mouse liver nuclei has been studied in situ using cytophotometric equipment. The accuracy and reproduciblity of the cytophotometer was ascertained by various procedures. 2. The absorption curve usually is represented by a primary peak at about 585 mµ and a slight secondary peak at about 550 mµ. Reduction or removal of nuclear protein by extraction or enzymatic procedures does not alter the shape of the curve. 3. Variations in hydrolysis times do not alter the shape of the curve obtained with nuclei fixed in Zenker's fluid. There are some differences indicated around the secondary peak of formalin-fixed tissue following different hydrolysis times. 4. Comparison of absorption curves of nuclei fixed in different fixatives shows negligible differences. 5. There are no differences in the absorption curves of nuclei isolated in one per cent citric acid and in nuclei isolated in 0.88 M sucrose.
A HISTOCHEMICAL INVESTIGATION OF NEUROSECRETORY SUBSTANCE IN THE RAT
Authors: HOWE A. , PEARSE A. G. E. 
Published: April 2011

(1) The chemical nature of `neurosecretory substance' has been examined by a variety of histochemical methods in the rat. (2) With the methods used it appears that this material is protein or polypeptide in character; the presence of lipid or carbohydrate is excluded, and it does not contain ribose nucleic acid. (3) These findings are discussed in relation to the theory of neurosecretion and the source of posterior lobe hormones.
THE DIFFERENCE IN PERMEABILITY OF CARTILAGE TO CATIONIC AND ANIONIC DYES
Authors: KANTOR THOMAS G. , SCHUBERT MANWELL 
Published: April 2011

It has been shown that fresh cartilage cubes are readily permeable to all of thirteen cationic dyes and one neutral dye tried. They are impermeable to thirteen of fourteen anionic dyes tried. This selective permeability of fresh cartilage to dyes can be altered by treatment which destroys the fixed negative charge of the cartilage. After this treatment, the cartilage cubes are readily permeable to anionic dyes. It is suggested that this membrane selectivity of cartilage may have physiological meaning.
THE ALKALINE PHOSPHOMONOESTERASE ACTIVITY OF CELL NUCLEI: ACTIVATION BY 0.16M MAGNESIUM
Authors: FEIGIN IRWIN , WOLF ABNER 
Published: April 2011

I. The presence of 0.16M magnesium chloride in the incubation mixture for the Gomori method for localizing alkaline phosphatase activity, markedly increases the enzyme activity in the nuclei of the guinea pig endometrium, ovary and adrenal cortex, rendering them visible after 5 minutes' incubation. The activity in cytoplasmic and extracellular sites is decreased. The staining of these same nuclei could be demonstrated by the Loveless and Danielli method of visualizing the phenolic component of a complex phosphate ester instead of precipitating the phosphate ions with calcium salts. The activity described may be specific for the tissues mentioned and serves as an instance in which nuclear staining in phosphatase preparations is clearly valid. II. The effect of magnesium and cyanide ions and other factors on the enzyme activity noted histochemically in nuclei and biochemically in nuclear cell fractions, was reviewed. The data support the validity of some earlier histochemical observations describing a nuclear phosphomonoesterase activity in many organs which differs from that in cytoplasmic and extracellular sites. They also suggest that the phosphatase activity in cell fractions of those biochemical preparations prepared in aqueous media are altered by diffusion and adsorption phenomena.
THE SPECIFICITY OF THE PERIODIC ACID-SCHIFF TECHNIQUE STUDIED BY A QUANTITATIVE TEST-TUBE METHOD
Authors: HOOGHWINKEL G. J. M. , SMITS G. 
Published: April 2011

A method is described for the quantitative estimation of periodic acid-Schiff positive substances in solutions of tissue components. It was found that hyaluronic acid and chondroitinsulphuric acid are periodic acid-Schiff negative. Any coloration observed could be ascribed to contaminating polysaccharides probably not or not exclusively composed of hexosamine and hexuronic acid. Unlike the hexosamine groups, the hexuronic acid groups in hyaluronic acid and chondroitinsulphuric acid are attacked by periodic acid without formation, however, of aldehyde groups. The chemistry of this process is discussed.
CYTOCHEMISTRY OF PINEAL LIPIDS IN RAT AND MAN
Authors: QUAY W. B. , ABRAMSON CONNIE 
Published: April 2011

Lipid cytochemistry studied in 130 rat pineals demonstrates two common types of parenchymal cells which may be involved in an endocrine secretory activity. The first cell type is characterized by its content of abundant lipid droplets, composed primarily of ethanol-soluble carbonyl lipids coated with phospholipid. The second cell type is distinguished by its content of a phospholipid cytoplasmic matrix containing few droplets on vacuoles. This cell type appears to be identical with the cell staining with orange G or phloxine in a modified chrome alum hematoxylin technique described previously. Parenchymal cells containing lipid occur in seven human pineals of diverse ages; and in the older pineals interlobular cells containing lipid occur as well. The lipid in human parenchymal cells differs from that in the type 1 parenchymal cells of the rat by, (1) forming smaller droplets, (2) containing a greater amount of neutral lipid and less carbonyl lipid, and (3) being associated with pigment fomation in adults and older people.