bluuebunny/crossref_metadata_2025_split huggingface dataset:
`SPREADING FACTOR', USED BEFORE FIXATION, AS AN AID TO CHOLINESTERASE LOCALISATION
Authors: WILLIAMS TERRY 
Published: April 2011

There is acknowledged difficulty in securing adequate detail in cholinesterase localisation, especially in sites of low activity. In addition, using the method of Koelle and Friedenwald, modifled by Coupland and Holmes, patchy or incomplete staining was often observed in a variety of tissues. Such preparations were unsuitable for histochemical assessment or for photography. It seemed likely that these shortcomings might be due, in part at least, to uneven fixation or uneven penetration of the fixative. In an attempt to correct or reduce these technical defects, tissues were infiltrated with testis hyaluronidase in isotonic saline, immediately after excision, and afterwards fixed and processed. It was found that hyaluronidase did not bring about diffusion of cholinesterase, as was originally feared. In a controlled series, using a number of tissues, hyaluronidase reduced the number of substandard preparations from 47 % to 6%. Improvement in reliability of the technique was achieved with some tissues, but not with others. Tissues or organs which with the ordinary method showed greatest patchiness seemed to benefit most from the hyaluronidase. It was suggested that speedy fixation with formol-saline can be facilitated with hyaluronidase and, contrasted with freeze-drying, the method has no major drawbacks.
A SPECIFIC STAIN FOR IRON USING 4,7-DIPHENYL-1,10-PHENANTHROLINE (BATHOPHENANTHROLINE)
Authors: HUKILL PETER B. , PUTT FREDERICK A. 
Published: April 2011

1. A new histochemical method for staining tissue iron is described, employing the chelating agent 4,7 - diphenyl - 1,10 - phenanthroline (Bathophenanthroline). 2. The method appears to be more sensitive than the classical Prussian blue method, and stains generally the same chemical forms of iron as does the latter. It is specific for iron. 3. The method has the theoretical advantage for histochemical purposes that, unlike the Prussian blue and Turnbull blue methods, it does not make use of an iron containing reagent. There is no possibility of a false positive reaction due to the reagent reacting with itself after oxidation by air or oxidizing agents in tissue. 4. It is recommended for investigative purposes and as a routine histologic stain where small amounts of iron are to be detected.
MORPHOLOGIC APPEARANCE AND CYTOCHEMICAL PROPERTIES OF MALIGNANT CELLS FOLLOWING RAPID DEHYDRATION
Authors: ORR HENRY C. , BELKIN MORRIS , HARDY WALTER G. , MERLER EZIO 
Published: April 2011

Cells of strains HeLa, D164, D189, D227 and P388 were grown on coverslips. The first four strains are of human cell origin; P388 is derived from a mouse leukemia. These coverslips were placed over phosphorus pentoxide in a desiccator which was rapidly exhausted, and permitted to remain so for an hour. Such dehydrated cells, examined with transmitted light and the phase contrast microscope, revealed considerable intact structural detail. The cell surface, nuclear membrane and cytoplasmic granules appeared well preserved. Adding a little distilled water to the dehydrated cells produced an improvement in visibility of many cellular components so that now such "restored" cells under transmitted light and phase contrast microscopy strikingly resembled living cells. When cytochemical tests were made for certain intracellular constituents, fat stained strongly with oil red O as did polysaccharide by the periodic acid-Schiff reaction. Giemsa also stained the cells well. Poor staining was obtained with hematoxylin and cosin, and with fast green, indicating adverse protein changes. In dehydrated cells, acid and alkaline phosphatase showed considerable activity, whereas cytochrome oxidase and succinic dehydrogenase appeared completely inactivated. The advantages of dehydration as a fixing procedure are (1) simplicity, (2) rapidity, (3) avoidance of damage due to chemical fixation, and (4) it permits storage for an indefinite period prior to further treatment.
THE CONNECTIVE TISSUE HISTOCHEMISTRY OF NORMAL AND SOME PATHOLOGICAL SKIN
Authors: SAMS W. MITCHELL , SMITH J. GRAHAM , DAVIDSON EUGENE A. 
Published: April 2011

A detailed histochemical study of the connective tissue of the skin has been presented. The stains used included hematoxylin and cosin, Mowry's colloidal iron and alcian blue for acid mucopolysaccharides, bromphenol blue for proteins, synthetic orcein and Verhoeff's for elastic tissue, and a combined Mowry's colloidal iron-orcein stain. Blocking procedures employed included methylation, demethylation, acetylation, deamination, and oxidation. Enzyme extractions were performed with testicular hyaluronidase. Representative diseases of the following three types were studied: 1) those due to or influenced by the effects of solar radiation; 2) those associated with abnormal elastic fibers without particular relationship to the amount of actinic radiation; and, 3) those associated with large numbers of fibroblasts. In chronically sun-damaged skin such as actinic elastosis, actinic keratosis, and basal cell epithelioma, markedly increased amounts of acid mucopolysaccharides were found in the areas of basophilic and orceinophilic material. Polymorphic light eruption and discoid and disseminate lupus erythematosus were found to have numerous acid mucopolysaccharide strands throughout the upper one-half of the dermis. Elastosis perforans serpiginosa displayed increased acid mucopolysaccharides associated with its orceinophilic fibers. Approximately 75% of the acid mucopolysaccharide in sun-damaged skin and elastosis perforans serpiginosa was removed by testicular hyaluronidase. In pseudoxanthoma elasticum each curled elastic fiber was found in the center of a pool of acid mucopolysaccharide, as though, when it retracted, it brought its mucopolysaccharide sheath with it. This acid mucopolysaccharide, in contrast to that found in sun-damaged skin and elastosis perforans serpiginosa, is only slightly labile to testicular hyaluronidase. Acid mucopolysaccharides were also found in large quantity wherever there were young fibroblasts as in dermatofibromas, neurofibromas, keloids, and fibroepithelial polyps, and is believed by the authors to represent de novo production by these cells rather than "mucinous degeneration." Hyaluronidase removed approximately 75% of the acid mucopolysaccharides in all conditions associated with young fibroblasts. The usefulness of the combined Mowry-Haleorcein stain was stressed. From the evidence presented, doubt was cast on the presently held concept that colloidal iron binding is by dissociated acid groups alone; iron may also be chelated by the intermolecular hydrogen bond of acid mucopolysaccharides. Oxidation greatly increased the intensity of Mowry-Hale staining: bromine probably by direct bromination and formation of cysteic acid and permanganate probably by formation of carboxyl groups.
ADENOSINE TRIPHOSPHATASE: A NEUROHISTOCHEMICAL STUDY
Authors: NAIDOO D. 
Published: April 2011

The location of adenosine triphosphatase in the brain has been studied in rapidly frozen-dried cerebral tissues of the Wistar rat. It is found that adenosine triphosphatase is an almost exclusively nuclear enzyme. Two tissue fractions of the cerebrum were separated, so that one sample was made up of vascular elements, and the other of neural elements. The two fractions were then studied for their adenosine triphosphatase activity, and compared with the histochemical findings. The two tissue fractions were found not to differ in the absence of bivalent cations. When Ca++ were added to the cerebral vascular suspension, ATPase activity was increased approximately 15 times, and only 3 times in the presence of Mg++. Conversely, the addition of Mg++ increased the ATPase activity of the neural fraction 200%; whereas, Ca++ was responsible for a 60% increase. This fact was detectable microscopically when Ca++ was found to intensify vascular nuclear staining, and Mg++ to increase the neuronal and glial nuclear staining. The results, histochemical and biochemical, are mutually confirmatory.
POLYSACCHARIDES OF CONNECTIVE TISSUE
Authors: DORFMAN ALBERT 
Published: April 2011

An attempt has been made to review the current status of knowledge of the chemistry and metabolism of the acid mucopolysaccharides of connective tissues. It has been pointed out that complex polysaccharides play a prominent role in the physiology of cell surface as well as the intracellular matrix. In mammalian tissues, connective tissues contain a variety of acid mucopolysaccharides with varying structure and biological activity. It is probable that their localization is correlated with their functional capacities. Rapidly progressing understanding of biosynthetic mechanisms indicates the pathways of biosynthesis of some members of this class of compounds although many important questions remain unsolved. It is apparent that the biosynthesis of these compounds is under control of a number of factors, most strikingly the endocrine agents.
THE INFLUENCE OF TECHNIQUE AND HORMONES ON HISTOCHEMICAL DEMONSTRATION OF ENZYMES IN INTESTINAL EPITHELIUM
Authors: LIU HSI-YEN , BAKER BURTON L. 
Published: April 2011

The influence of technical variations and of hypophysectomy and adrenalectomy on the demonstration of enzymes in the jejunal epithelium was studied. The application to fresh frozen sections of procedures for cytochrome c oxidase, succinic dehydrogenase, diphosphopyridine nucleotide diaphorase and lactic dehydrogenase failed to reveal a precise mitochondrial localization, basing the conclusion on comparison with mitochondria as demonstrated by staining of Regaud-fixed mucosa with the Altmann-Masson procedure. Fixation in cold formalin prior to freezing and sectioning permitted a more satisfactory localization of succinic dehydrogenase in mitochondria. Hypophysectomy or adrenalectomy resulted in a reduction in number, size, staining and orderly arrangement of mitochondria in the villous epithelium as revealed in cytological preparations and those fixed and stained for succinic dehydrogenase. Alkaline phosphatase activity was also depressed. The intensification of cytoplasmic staining for lactic dehydrogenase, diphosphopyridine nucleotide diaphorase and succinic dehydrogenase in fresh frozen tissue probably represents concentration of formazan by reduction in cell volume and not increased enzyme activities.
ELECTRO- AND IMMUNOELECTROPHORETIC PATTERNS IN THE LARVAL SALIVARY SECRETION OF CHIRONOMUS THUMMI
Authors: KATO K.-I. , PERKOWSKA E. , SIRLIN J. L. 
Published: April 2011

The salivary secretion of chironomid larvae was studied electro- and immunoelectrophoretically. The secretion could be resolved into 14 nigrosin-positive (protein) bands and 10 metachromatic (mucopolysaccharide) bands in the pherograms. Much less stainability was found in some of these bands in the pherograms of the gland cell components. The polysaccharide bands roughly corresponded with an equal number of protein bands at the same positions in the pherogram. An antiserum against a cell extract of whole larvae (excluding the salivary glands) yielded 6 groups of precipitation crescents on the pherogram of the cell extract. The same antiserum which had been previously absorbed with the salivary secretion yields a reduced number of crescents, specially on the anodic side of the pherogram.
DEHYDROGENASES IN DEVELOPING TEETH OF RATS
Authors: FULLMER HAROLD M. 
Published: April 2011

Odontogenesis was studied in unfixed cryostatcut sections of 5-day old rats with histochemical methods for the demonstration of succinic, malic, isocitric, glutamic, lactic, β-hydroxybutyrate, α-glycerophosphate, glucose-6-phosphate and 6-phosphogluconate dehydrogenases and DPN and TPN diaphorases. Except for weak glucose-6-phosphate and 6-phosphogluconate dehydrogenases, odontogenic tissues (particularly ameloblasts, odontoblasts and stratum intermedium cells) exhibited moderate activity indicating a broad metabolic behaviour of these cells. Cells of the stratum intermedium and the stellate reticulum display an enzymatic activity parallel to ameloblasts which suggests that they participate actively in amelogenesis. Tetranitro-BT and Nitro BT appear to give definitive localization of dehydrogenase activity.
VARIATIONS IN THE ELECTROPHORETICALLY SEPARATED ACID PHOSPHATASES OF TETRAHYMENA
Authors: ALLEN SALLY LYMAN , MISCH MARGARET SEGUR , MORRISON BARBARA M. 
Published: April 2011

The acid phosphatases of variety 1 of Tetrahymena pyriformis can be separated into 17 zones by electrophoresis in starch gels of pH 7.5. All of these acid phosphatases have an optimal pH of about 5.0 and are inhibited by 10 mM sodium fluoride on d-tartaric acid. With one exception, all hydrolyze sodium α-naphthyl acid phosphate. Differences between the acid phosphatases are observed in their ability to hydrolyze other substrates using either the coupling technique or a modification of the Gomori-lead method. Inhibition of 2 of the acid phosphatases occurs in the presence of 1 mM of Mn++ or Zn++; 0.1 mM of p-chloromercurobenzoic acid inhibits these and others. Variations between the acid phosphatases were observed under different growth conditions and in their distribution in various cell fractions. A major variation in the acid phosphatases that is under genetic control occurs in extracts of different genotypes. The results suggest that the electrophoretically separated acid phosphatases are a family of enzymes that vary in their degree of relationship. Some are different enzymes. Others are more closely related and represent variations of a single enzyme; i.e., mutant forms produced by different alleles, hybrid forms produced by interaction of alleles, and isozymes produced by a single gene. Most of the acid phosphatases are probably associated with lysosomes, although one appears to be associated with microsomes. This acid phosphatase is much more active in synthetically grown cells than in cells grown in proteose-peptone or in bacterized medium. It does not hydrolyze sodium β-glycerophosphate.
HISTOCHEMICAL EVIDENCE OF PROTEOLYSIS IN BONE: THE INFLUENCE OF PARATHORMONE
Authors: BÉLANGER LEONARD F. , MIGICOVSKY B. B. 
Published: April 2011

Proteolytic activity has been revealed at the level of the large osteocytes of young chick and rat bone by digestion of gelatin in a photographic emulsion substrate. The reaction was enhanced by pretreatment of the animals with parathyroid extract. The reaction was inhibited by addition to the incubating medium of sodium fluoride or potassium ferrocyanide. It was also inhibited by fixation in ethanol-formaldehyde acetic acid. The present results seem to confirm and explain further the resorptive potential of the mature osteocytes.
HISTOCHEMICAL AND QUANTITATIVE CHANGES IN GLYCOGEN AND PHOSPHORYLASE DURING DISUSE ATROPHY OF THE PIGEON PECTORALIS
Authors: VALLYATHAN N. V. , CHERIAN K. M. , GEORGE J. C. 
Published: April 2011

Histochemical and quantitative biochemical changes in glycogen content and phosphorylase activity of pigeon breast muscle were studied during development of disuse atrophy from one day to sixty days. Of the two types of muscle fibers present in the pigeon breast muscle the large white glycogen-loaded fibers, which are well adapted for an anaerobic metabolism, were found to lose their glycogen store and phosphorylase activity during the early days of atrophy. On the other hand, the smaller, red, fat-loaded fibers, which are well adapted for an aerobic metabolism, were found to acquire more glycogen and phosphorylase activity during the same period. The staining reaction of these fibers for phosphorylase thus, as with biochemically different branching enzymes, presented a mosaic. However, a few small fibers showed no increase in glycogen content or phosphorylase activity. The histochemical observations also revealed that the majority of the large white fibers were reduced in size, filled with fat, and separated by connective tissue during the later stages of atrophy. Fibers of this sort disappeared or were completely engulfed by connective tissue. Very few small fibers changed in this way. In certain regions an entire fasciculus containing both types of fiber was found to be replaced by fat and connective tissue, leaving a few narrow red fibers intact. There was no phosphorylase activity in the connective tissue. The quantitative studies showed a decrease in glycogen and phosphorylase levels in the muscle early in disuse. After 60 days of atrophy the phosphorylase level recovered somewhat but the glycogen level remained low.
CYTOCHEMICAL DEMONSTRATION OF ACID PHOSPHATASE IN HEMATOPOIETIC CELLS IN HEALTH AND IN VARIOUS HEMATOLOGICAL DISORDERS USING AZO DYE TECHNIQUES
Authors: KAPLOW L. S. , BURSTONE M. S. 
Published: April 2011

Acid phosphatase activity in the formed elements of blood and bone marrow has been demonstrated using unfixed smears and azo dye coupling techniques with naphthol AS-MX or AS-TR phosphates as substrates. A similar staining pattern was also obtained using a new substrate, hydroxycarbazole phosphate. When this compound was used, hydrolysis was more rapid and good staining was obtained using smears fixed at room temperature (25°C) in 60% acetone buffered to pH 4.2-4.5 with citrate. Such preparations showed good cytological detail; however, the azo dye is less chromogenic than that formed with the substituted naphthol phosphates and nonspecific staining due to free hydroxycarbazole was observed. The use of ethylene diamine tetraacetic acid (EDTA) as an anticoagulant did not interfere with the reaction. Moderate reduction in staining intensity was observed using smears stored at room temperature longer than 24 hours prior to incubation. Dye precipitation was observed in most cells and was strongest in bone marrow osteoclasts and phagocytic reticulum cells. Immature myeloid cells showed moderately strong activity and this was particularly evident in peripheral blood smears of some patients with untreated chronic myelogenous leukemia. Increased staining was also observed in the plasma cells of 2 patients with untreated multiple myeloma. No consistent deviation from the normal was observed in the remaining disorders studied.
ELECTROPHORETIC EVIDENCE FOR ISOZYMES OF ARYLAMIDASE ("AMINOPEPTIDASE") IN THE RAT. I. RENAL, SERUM AND URINARY ENZYMES SPLITTING dl-ALANYL-2-NAPHTHYLAMIDE AND l-LEUCYL-2-NAPHTHYLAMIDE
Authors: MONIS BENITO 
Published: April 2011

Bilateral nephrectomy elicited no changes of serum blood levels of arylamidase assayed with two chromogenic substrates (l-leucyl-2-naphthylamide and dl-alanyl-2-naphthylamide) two days after the operation. Since rat urine contains similar enzymes it was postulated that the urinary enzyme is of renal origin and distinct from serum arylamidase. For this purpose, electrophoretic studies were undertaken. Starch block (for quantitative determinations) and starch gel for zymograms of arylamidase were used. It was shown that both procedures demonstrated the identity of urinary and renal arylamidase, which was distinct from the serum enzyme. The renal and urinary enzyme showed two distinct isozymes: one remaining at the origin and the other migrating somewhat less than the isozyme in serum. It is postulated that the isozyme remaining at the origin corresponds to a membrane-bound form, whereas the electrophoretically-mobile isozyme is found in the cellular supernatant fraction. The arylamidase from serum was represented by a single enzyme which migrated farthest towards the anode. Histochemical procedures for the demonstration of arylamidase activity in tissues at the light microscopic level permit the localization of enzyme(s) that can hydrolyze synthetic chromogenic naphthylamides containing l-leucyl and dl-alanyl groups (1). Rat kidneys have a high concentration of histochemically demonstrable arylamidase in the proximal convoluted tubules (2). Blood levels of arylamidase in the normal adult rat vary within a narrow range (3). The observation of arylamidase activity in rat urine raised several questions that led to the studies which are the basis of the present report. The relationship of serum and urinary arylamidase was the starting point of this investigation. It was speculated that if the urinary enzyme had its origin in the serum, bilateral nephrectomy should alter blood levels of the enzyme. If no such changes occurred, this would suggest that the urinary enzyme was released from the proximal convoluted tubule. In fact, total renal ablation led to no significant variation of serum arylamidase activity. It was therefore postulated that urinary arylamidase originated in the kidneys and that both are distinct from serum arylamidase. To test this hypothesis, zone electrophoretic studies were undertaken using two different supporting media. It will be shown that distinct molecular forms or isozymes of arylamidase can be separated from rat tissues and fluids by differential electrophoretic mobility. The electrophoretic identity of urinary and renal arylamidase, both of which are distinct from the serum arylamidase, is demonstrated in this study.
ON THE BINDING OF DIRECT COTTON DYES BY AMYLOID
Authors: PUCHTLER HOLDE , SWEAT FAYE , KUHNS JAMES G. 
Published: April 2011

Application of 30 direct cotton dyes in aqueous salt solution to amyloid in human tissues yielded different staining patterns. Only dyes with good wool reserve without reactive metal atoms, i.e., dyes which stain cellulose but not proteins or polyamides, colored amyloid selectively without differentiation. Direct dyes for union fabrics, i.e., dyes which color cellulose and protein fibers, stained all tissue structures. The effects of deamination and acetylation on the binding of direct dyes with good wool reserve indicate interaction of these dyes with hydroxyl groups of amyloid. The increase in intensity of staining upon addition of NaCl to the dye solution suggests a nonionic type linkage. According to chemical data on the direct dyeing of cellulose, direct dyes are adsorbed by hydrogen bonding between hydroxyl groups of the polysaccharide chains and suitable groups in the dye. It seems probable that a similar mechanism is involved in the binding of these dyes by amyloid. The selective staining of amyloid with direct dyes with good wool reserve, which do not color proteins, suggests that the polysaccharide moiety of amyloid may play a major role in the binding of direct dyes.
THE STAINING OF TRIPHOSPHATASES BY THE CHELATE REMOVAL METHOD
Authors: BERG GEORGE G. 
Published: April 2011

In the histochemical staining of enzymes by the chelate removal method, the substrate is a chelate, and the enzyme unmasks bound ions which are precipitated by suitable capturing ions in the incubating mixture. The new method allows the staining of phosphatases which act on substrates that are insoluble in the Gomori mixtures. Three inorganic polyphosphatases were stained in the first application of the chelate-removal principle: neutral triphosphatase, alkaline triphosphatase, and alkaline pyrophosphatase. The incubating media had lead-polyphosphate chelates as substrates, with "tris" or barbital buffers. Orthophosphate was added to some solutions as an auxiliary capturing anion. Alkaline triphosphatase as well as alkaline pyrophosphatase was demonstrated in the columnar epithelium of duodenal mucosa of mouse and rat, alkaline triphosphatase in the convoluted tubules of kidney, and neutral triphosphatase in bile canaliculi of liver. The properties of the three enzymes were examined, and the relation of the stains to biochemically defined enzymes of the inorganic polyphosphatase group was discussed. The chelate removal method was also used for the histochemical demonstration of adenosine tri- and diphosphatase. Evidence for this mechanism of ATP-ase staining is presented as an addendum.