bluuebunny/crossref_metadata_2025 huggingface dataset:
A COMPARATIVR STUDY OF ENZYMATIC STAINING REACTIONS IN THE RAT KIDNEY WITH NECROBIOSIS INDUCED BY ISCHEMIA AND NEPHROTOXIC AGENTS (MERCUHYDRIN AND DL-SERINE)
Authors: WACHSTEIN M. , MEISEL E.
Published: April 2011

Enzymatic staining reactions for the demonstration of succinic dehydrogenase, DPN diaphorase, esterase, glucuronidase, nonspecific alkaline and acid phosphatase, phosphatase at pH 7.2, glucose-6-phosphatase, 5-nucleotidase and adenosine triphosphatase were applied to rat kidneys in which necrobiosis was induced by either the ligation of renal vessels or the administration of the nephrotoxic substances dl-serine and Mercuhydrin. Frozen sections of fresh and formalin-fixed tissues were used throughout. The distribution patterns of these enzymes found in the normal rat kidney were consistent and in agreement with previous observations. The staining pattern of adenosine triphosphatase at pH 7.2 was distinetly different from that of 5-nucleotidase and nonspecific phosphatase.

Following occlusion of the blood supply, necrobiotic changes developed relatively slowly. An observation apparently so far not recorded in the literature, is the occurrence of protruded proximal convoluted tubules in the lumen of Bowman's capsules. Nephrotoxic drugs lead to a rapidly developing coagulation necrosis in proximal convoluted tubules within a few hours. Without exception, diminution of enzymatic activity occurred later in ischemic as compared to toxic necrosis.

Definite differences in the histochemically demonstrable enzymes were noticed in cells undergoing necrobiosis. Non-specific alkaline and acid phosphatase and phosphatase at pH 7.2, esterase and 5-nucleotidase remained at least partially active in cells made necrotic by all procedures. The oxidative enzymes succinic dehydrogenase and DPN diaphorase were more completely inactivated by toxic as compared to ischemic necrosis. The reverse was true for adenosine triphosphatase, glucuronidase and glucose-6-phosphatase. Of all enzymes tested, succinic dehydrogenase was the most sensitive indicator of nephrotoxic cell damage, since distinct inactivation occurred with both dl-serine and Mercuhydrin within 30 to 60 minutes.
THE DISTRIBUTION OF CHOLINESTERASE IN THE SUBMAXILLARY AND SUBLINGUAL SALIVARY GLANDS OF THE RAT
Authors: SNELL R. S. , GARRETT J. R.
Published: April 2011

1. The distribution of cholinesterase in the submaxillary and sublingual salivary glands has been studied histochemically in 24 mature rats.

2. A small amount of true cholinesterase was found in the nuclei and cytoplasm of the duct cells but was absent from the acinar cells.

3. True cholinesterase was present in the nerve trunks accompanying the intra and interlobular ducts of both glands but the concentration was less in the nerves supplying the sublingual gland.

4. True cholinesterase was also found to be present in the form of a fine network around the acini of both glands but was much less evident around the acini of the sublingual gland.

5. The significance of this cholinesterase network in relation to the mode of termination of the nerve fibres to the different secretory cells of the glands is discussed.
REDUCING SITES IN PACTERIAL CELLS OF THE ANAEROBIC GENUS CLOSTRIDIUM AND THEIR DIFFERENTIATION FROM OTHER CELL STRUCTURES
Authors: DAVIS JOHN C. , MUDD STUART
Published: April 2011

Granular sites of oxidation-reduction are readily demonstrable in the cells of bacteria of the genus Clostridium, using either intravital redox indicators or phospholipid stains. These granules in Clostridium do not give the reactions for cytochrome oxidase. In other respects their tinetorial reactions and distribution resemble those of the oxidation-reduction sites of other bacteria, which have been defined as bacterial mitochondria (2). The reducing granules are differentiated from cell-walls and intracellular septa, nuclei, iogen granules and spores. During the process of sporulation reducing granules are demonstrable within the cytoplasm of the spore bearing cell, and in particular adjacent to the outer border of the spore. With maturation of the spore and disappearance of most or all the cytoplasm of the mother-cell, reducing granules could no longer be demonstrated.
METAL REDUCTION REACTIONS OF The MELANINS: SILVER AND FERRIC FERRICYANIDE REDUCTION BY VARIOUS REAGENTS IN VITRO
Authors: LILLIE R. D. , HENSON JACQUELINE P. GRECO , BURTNER HELEN C. JACQUIER
Published: April 2011

Test tube reactions of 163 reagents with silver solutions are recorded in tabular form. The silver solutions used were silver nitrate at pH 4 and at pH 6, ammoniacal silver nitrate, all at about 25°C., and Gomori's methenamine silver at 60°C. Ferric ferricyanide reduction is also included in the tables for comparison.

Of the substances tested 30 quite promptly reacted with acid silver nitrate as well as the other reagents, 28 gave delayed reactions with acid, prompt with alkaline silver, 24 gave variable acid, and delayed alkaline silver reactions, 27 gave no reaction with acid silver and delayed reactions with alkaline, 16 reacted slowly with hot methenamine silver and not or scarcely at all with cold ammoniacal silver. The remaining 38 reactants did not blacken with silver solutions.

Also tabulated are the pigment formation reactions of 13 polyphenols, indoles and sulfur amino acids with chromic acid, potassium bichromate, potassium chromate, periodic acid, sodium iodate and potassium chlorate.
TRICHOXANTHIN, THE YELLOW GRANULAR PIGMENT OF GUINEA PIG HAIR FOLLICLES AND HAIRS
Authors: LILLIE R. D.
Published: April 2011

Trichoxanthin is a granular yellow pigment occurring in hair matrix and hair medulla of yellow and red guinea pig skin. It is insoluble in organic and dilute mineral acids, water and organic solvents, irregularly soluble in dilute alkali and promptly removed by methylation though not by acidified higher aliphatic alcohols.

It reduces ferric ferricyanide and ammoniacal silver promptly, acid silver nitrate slowly. Indole reactions, azo-coupling reactions and Gibbs's phenol reaction are negative.

It is strongly basophil at pH 1 and binds ferrous ions, in these characters resembling melanin.

It is more promptly bleached by HNO3, CrO3, KMnO4 and CH3CO3H than is cutaneous melanin of the same species and its reduction reactions are more rapidly affected by oxidants.

Reduction of acid silver nitrate is not appreciably accelerated by prior hydrosulfite reduction, in distinction from melanin.

On the basis of the modifications of the metal reduction reactions by oxidants and reductants, the possibility of a p-quinhydrone structure is suggested.
DISTRIBUTION OF PHOSPHAMIDASE ACTIVITY IN THE MALE ALBINO RAT
Authors: MEYER J. , WEINMANN J. P.
Published: April 2011

Phosphamidase activity was studied in organs and tissues of the rat, using the modification of Gomori's histochemical technique suggested by Meyer and Weinmann. Enzyme activity was allowed to go on for periods varying between 1 and 3.5 hrs. Earliest appearance, rate of increase, reproducibility and cellular distribution of staining were observed. Sites of high activity (visible precipitation in 1 to 1.5 hrs. of enzyme action) were found in epithelia and nerve tissue. High phosphamidase activity in epithelia occurred in the cells specialized for active transfer and other types of physico-chemical work, in exocrine and endocrine glands synthesizing nonprotein compounds and in tissues undergoing histodifferentiation in the adult animal. The sites of high enzyme activity in nerve tissue were found to be neuropil and nerve processes of the central nervous system and the cytoplasm of autonomic ganglion cells and nerve processes of the peripheral nervous system.

Phosphamidase activity was found absent in 3.5 hr. periods of enzyme action from epitheial cells specialized for passive transfer, from unspecialized cells and from many glands synthesizing proteins, further from the cell bodies of the cells in the central nervous system and from muscle of all types. Comparison of distributions shows phosphamidase activity demonstrably distinct from acid and alkaline phosphomonoesterase for a number of organs. Comparison of the distributions shows the sites of high phosphamidase activity to be in those tissues which do not respond to insulin stimulation; conversely, the tissues in which transfer of simple sugars is mediated by insulin do not show phosphamidase activity. A role in energy metabolism complementing the action of insulin is tentatively suggested for phosphamidase.
THE FEULGEN-DEOXYRIBONUCLEIC ACID ABSORPTION CURVE IN NORMAL AND TUMOROUS TISSUES
Authors: KASTEN FREDERICK H.
Published: April 2011

Absorption curve analysis of the Feulgen-deoxyribonucleic acid dye complex in situ fail to reveal any curve differences in nuclei of normal and tumorous tissues of the human, hamster, and mouse.
RADIOAUTOGRAPHIC DEMONSTRATION OF NORADRENALINE AND ADRENALINE IN THE ADRENAL MEDULLA WITH I131-LABELLED IODATE AND IODATE-FORMALIN
Authors: ERÄNKÖ OLAVI
Published: April 2011

Adrenals of the hamster, the mouse and the rat were studied.

Radioactive iodate solution was prepared by mixing carrier-free I131 with a saturated solution of potassium iodate. In adrenals immersed in this solution for 3-96 hours and subsequently fixed in 10 per cent formalin, a typical brown reaction was seen in some, presumably noradrenaline-containing, cells of the medulla. Radioautographs prepared of the same sections indicated that the distribution of the brown color and that of the radioactivity were identical. This suggests that the brown color produced by treatment with iodate is due to an iodine-containing compound. The cortex exhibited only a weak radioactivity and no color.

Adrenals immersed in a mixture of 1 volume of commercial formalin and 9 volumes of the radioactive iodate solution for 12-96 hours showed a brown color in all medullary cells. The medulla was demonstrated by radioautographs to contain large quantities of iodine. However, the cortex was also strongly radioactive in spite of previous treatment with an alcohol-ether mixture.

Some observations on the chromaffin reaction were also reported.
CYTOCHEMICAL DEMONSTRATION OF SUCCINIC DEHYDROGENASE BY THE USE OF A NEW p-NITROPHENYL SUBSTITUTED DITETRAZOLE
Authors: NACHLAS MARVIN M. , TSOU KWAN-CHUNG , DE SOUZA EUSTACE , CHENG CHAO-SHING , SELIGMAN ARNOLD M.
Published: April 2011

1. A new p-nitrophenyl substituted ditetrazole, 2,2'-di-p-nitrophenyl-5,5'-diphenyl-3,3'-(3,3'-dimethoxy-4,4'-biphenylene) ditetrazolium chloride (Nitro-BT), is introduced, which possesses most of the properties of an ideal reagent for histochemical purposes. It permits the cytochemical visualization of the sites of enzymic activity in tissue sections from 2-8 µ thick, under aerobic conditions, after a relatively short incubation period.

2. A comparison was made of the various tetrazoles in common use with several new p-nitro substituted analogues of these agents.

3. A survey was presented of the enzyme distribution in the major tissues of the rat and dog. Evidence for the reliability of cytochemical localization is provided by illustrative material in selected tissues.

4. The new tetrazole can be employed for the diphosphopyridine nucleotide and triphosphopyridine nucleotide diaphorase systems as well as for the succinic dehydrogenase system by substituting Nitro-BT for blue tetrazolium (BT) in the incubation media.
PRESENCE OF CARBOHYDRATES WITH FREE 1,2-GLYCOL GROUPS IN SITES STAINED BY THE PERIODIC ACID-SCHIFF TECHNIQUE
Authors: LEBLOND C. P. , GLEGG R. E. , EIDINGER D.
Published: April 2011

The mechanism of periodic acid (PA) -Schiff staining was investigated by analysis of materials isolated from connective tissue derivatives and secretory products which in sections stain to a variable extent with the PA-Schiff technique. The methods devised for the isolation of these two classes of materials involved extraction with alkali and saline respectively, followed by precipitation with ethanol.

Chemical determinations reveal the presence of hexose, hexosamine, methylpentose, nitrogen, and perhaps sialic acid in these materials (Table 3). The hexose content parallels the intensity of PA-Schiff staining (Tables 1 and 2). The monosaccharide components detected by paper chromatography are galactose and fucose, mannose in most cases and occasionally glucose and rhamnose, but not uronic acid . The individual monosaccharide components of each material differ in a quantitative as well as qualitative manner which could be reduplicated on repeated extractions, a fact considered to be presumptive evidence of the homogeneity of the isolated substances.

After treatment of the materials with periodic acid, all glucose and fucose residues and most galactose and mannose residues are eliminated; and, therefore, these monosaccharides must be present in such a form that their 1,2-glycol groups are free for oxidation with periodic acid.

Since all PA-Schiff reactive materials under investigation contain bound galactose and fucose, as well as hexosamine and probably sialic acid, and are associated with protein, they make up a distinct class of substances, referred to as carbohydrate-protein complexes. It is proposed that, once glycogen has been removed from sections with amylase, the PA-Schiff technique is specific for the carbohydrate-protein complexes thus defined.
A CHEMICALLY SPECIFIC TECHNIQUE FOR THE INTRACELLULAR LOCALIZATION OF INORGANIC PHOSPHATE
Authors: TANDLER C. J.
Published: April 2011

The chemical specificity of the lead acetate reaction (17) has been improved by the use of ammonium acetate and other variants in the cytochemical technique. It has been possible to establish that inorganic phosphate is a normal component of the nucleolus.

Whether it is present only in the nucleoli, as suggested by the cytological specificity of the test, or whether it represents only a fraction of the total inorganic phosphate of the cell, remains to be proved.
PROBLEMS OF ENZYMATIC LOCALIZATION BY CHEMICAL REACTIONS APPLIED TO TISSUE SECTIONS
Authors: NACHLAS MARVIN M. , YOUNG ANITA C. , SELIGMAN ARNOLD M.
Published: April 2011

1. During the preparation of the tissues for histochemical studies, attention must be directed toward the conservation of enzymatic activity and the maintenance of enzyme localization. No matter which technique is employed, the histological manipulation can result in either false positive or false negative enzyme localization.

2. The special characteristics of the chemical reactions used for the demonstration of enzymes in tissue sections have been listed, and an analysis of the desired properties of each constituent has been made.

3. When the tissue sections are introduced into the incubation medium new problems arise which are related to the properties of both the enzyme being studied and the chemical reagents being used.

4. Theoretical arguments have been presented to indicate that the discrete deposition of the dye within the cell does not necessarily represent a more accurate localization for all enzymes, than does the widespread dispersion of the dye throughout the cells.

5. Although we have some understanding of the ideal properties of the substrate, the split product, and the final pigments, there are many situations where these cannot be attained. Nevertheless, by the selection of the proper combination of suboptimal conditions, an accurate localization of many enzymes can be achieved.
HISTOCHEMICAL OBSERVATIONS ON THE FETAL OVARY AND TESTIS OF THE HORSE
Authors: DAVIES J. , DEMPSEY E. W. , WISLOCKI G. B.
Published: April 2011

A fetal horse ovary at the 6th month of gestation and a fetal ovary and testis at the 9th month have been studied histochemically with special reference to the cytoplasmic lipids of the interstitial cells. At least two lipid fractions appeared to be present, one which was soluble in acetone and was mainly responsible for the positive "plasmal" reaction, and another, insoluble in acetone, which was responsible for the sudanophilic, Ashbel-Seligman and periodic acid-Schiff positive material remaining after acetone extraction. The interstitial cell lipids in the older ovary and testis were also autofluorescent both before and after acetone extraction. No birefringent material was noted in any of the specimens.

The tentative interpretation of the findings as indicative of possible sites of ketosteroid hormone synthesis or storage is discussed briefly and related to the problem of the hormonal interrelationships of mother and fetus in the horse.
THE HISTOCHEMICAL DEMONSTRATION OF MONOAMINE OXIDASE ACTIVITY BY TETRAZOLIUM SALTS
Authors: GLENNER G. G. , BURTNER HELEN J. , BROWN G. W.
Published: April 2011

1. By the use of tryptamine as substrate in the presence of tetrazolium salts in a solution buffered to pH 7.6 histochemical localization of monoamine oxidase activity is achieved during a 45 minute incubation of tissue sections at 37°C.

2. Inhibition experiments implicate the postulated acetaldehyde product of monoamine oxidase action on tryptamine and serotonin in the reduction of the tetrazoles. It was not determined whether this reduction is nonenzymatic or mediated through a flavoprotein enzyme system.

3. Localization of the formazan to autonomic ganglia and fibers, chief cells of the gastric fundus, mucosal epithelium of the duodenum and certain specific zones of the renal tubules by means of the tryptamine-tetrazolium technique was confirmed as indicating monoamine oxidase activity by identical localization using the hydrazone precipitation technique of Koelle and Valk.
AZO-COUPLING REACTIONS OF PROTEIN-BOUND AMINO GROUPS IN PLANT TISSUE BY OXIDATIVE DEAMINATION METHOD
Authors: SURREY KENNETH
Published: April 2011

Fresh freehand bean cotyledon sections were killed and fixed in chilled 70% alcohol, hydrolysed with 1% NaOH solution, and washed in a mixture of chloroform-methanol. Suspended the sections in 5% hydrazine hydrochloride solution in pH 6-7 phosphate buffer and passed CO2 slowly to block free aldehyde groups. Washed and deaminated oxidatively with 1% chloramine-T in pH 7.5 phosphate buffer. The protein-bound aldehydes obtained by the above procedure were demonstrated with (a) Schiff's reagent or malachite green (b) arylhydrazines and (c) arylamines. Among arylhydrazines (1) azobenzenephenylhydrazine sulfonic acid, (2) 2,4-dinitrophenylhydrazine and (3) 3-hydroxy-2-naphthoic acid hydrazide were used. Compound (1) gave a purple dye with protein-bound aldehydes. A hydrazone formed after the condensation of the reagent (2) with protein-bound aldehyde, was reduced, diazotized and coupled with H-acid to produce a red-purple dye. After condensation the reagent (3) was coupled with the stable diazotate of 5-nitro-o-anisidine to produce a bright red dye. Among arylamines, H-, Chicago-, J-, and gamma-acids were used. These acids after having condensed their —NH2 groups with aldehydes were coupled with the stable diazotate of 5-nitro-o-anisidine to produce red dyes.
HISTOCHEMICAL METHODS FOR CALCIUM
Authors: McGEE-RUSSELL S. M.
Published: April 2011

An account is given of an investigation of histochemical methods for calcium using standard test tissues (McGee-Russell, 1957) known to contain calcium salts, as test objects upon which to assess the success, reproducibility, and mutual consistency of methods. The methods for the revelation of calcium in the literature fall into two groups: metal substitution techniques, and dye-lake reactions. Published methods have been tested and many have been found of little use for cytological investigations. A set of six metal substitution techniques, including a new copper dithioxamide method, has been established as giving useful results. The empirical basis of the staining reactions for calcium with dyestuffs has been investigated, and sensitive and specific techniques using anthraquinone derivatives are outlined. The significance of pH in dye-lake reactions is emphasized, and it is pointed out that previous results with anthraquinone dyestuffs have been achieved by an empirical adjustment of pH in the staining solution. The necessity of making a distinction between the formation of a dye-lake precipitate as a true histochemical positive, and staining effects which may occur upon tissue components in the neighborhood of calcium deposits, through the interaction of the dye, the dye-lake, the calcium salt, and the matrix, is stressed.

An oxazin dyestuff, gallamine blue, recently introduced for the staining of calcium deposits, was found to be inconsistent in its results, and capable of giving misleading information. Its general use is not recommended. The possibility of studying calcium deposits by means of the presence of contaminating metals is considered. The reagent sodium rhodizonate was found to give very fine localization of known calcium deposits in a number of tissues.
HISTOLOGICAL LOCALIZATION OF GLYCOGEN IN THE URINARY TRACT AND LUNG
Authors: SPICER S. S.
Published: April 2011

Glycogen in normal rabbit kidney is confined to the collecting tubule epithelium. The amount present increases from infrequent particles in the cortex to heavy deposits at the papilla. Only Bouin's fluid adequately preserves this glycogen, but that in the epithelium and smooth muscle of the renal pelvis and ureter and the bronchiolar epithelium of rabbits may be demonstrated after a variety of fixatives. Monkey kidney reveals glycogen also in the distal straight tubules. Rats and hamsters show little glycogen in renal tubules and mice and dogs none. 24 hours fasting had no effect on the bronchiolar or the collecting tubule glycogen of rabbits but alloxan hypoglycemia depleted the latter completely. Rabbits with alloxan hyperglycemia showed abnormal deposits in the epithelium of the distal straight and convoluted tubules. From available histochemical knowledge, the enzyme structure of the collecting duct is compatible with glycogen accumulation in that part of the duct where energy requirements are low. Glycogen deposition in the proximal and distal tubules in diabetes appears incompatible with the enzyme structure normally present in these segments.
THE LOCALIZATION OF ADENOSINE TRIPHOSPHATASE IN LIVER: IN SITU STAINING AND CELL FRACTIONATION STUDIES
Authors: NOVIKOFF ALEX B. , HAUSMAN DAVID H. , PODBER ESTELLE
Published: April 2011

1. In the Padykula-Herman staining medium, only 20% of the adenosine triphosphatase (ATP-ase) activity of rat liver homogenate is manifested, because of inhibition by the high level of calcium.

2. In the Wachstein-Meisel staining medium, only 15% of the ATP-ase activity is manifested, because of the high lead to ATP ratio.

3. The distribution patterns of the residual activity among the isolated subcellular fractions show it to be ATP-ase rather than non-specific phosphatase.

4. Cold formol-calcium fixation (33½-23 hours) reduces the adenosine triphosphatase activity of pieces of liver by 85-90%. The reduction is somewhat less when suspensions (homogenates) are fixed. The loss in activity due to fixation is much greater for the mitochondrial than for the nuclear fraction.

5. We have found the unmodified Wachstein-Meisel technique to give more reliable and consistent staining than any other method tried.

6. Chemical data show the mitochondria to be the major site of ATP-ase activity in rat liver; staining results are not inconsistent with this finding.

7. Staining results show the ATP-ase activity of the nuclear fraction to reside in the bile canaliculi and mitochondria present in the fraction.

8. There is no evidence that the nuclei of rat liver possess adenosine triphosphatase; 5'-nucleotidase activity is indicated with the calcium staining procedure at alkaline pH, but not with the lead procedure at pH 7.2.

9. The possible significance of high ATP-ase activity in the bile canaliculi and portal vessels is considered briefly.
A CYTOLOGICAL BASIS FOR THE BIOCHEMICAL STUDY OF BRONCHIAL EPITHELIUM
Authors: SPENCER BRIAN
Published: April 2011

A technique is described for the removal of epithelium from autopsy specimens of human bronchus and trachea so as to give a cell suspension in 0.25 M sucrose containing 0.01 M ethylenediamine tetraacetate, pH 7.4, which is suitable for biochemical study. The technique ensures that the suspension contains the whole of the epithelium from the treated area and that it is uncontaminated with mucus or connective tissue. Total and differential cell counts enable a correlation to be made between biochemical assays and the cytological make-up of the tissue.
HISTOCHEMICAL DEMONSTRATION OF AN ACETYLCHOLINESTERASE IN THE OVA OF SCHISTOSOMA MANSONI
Authors: PEPLER W. J.
Published: April 2011

1. An acetylcholinesterase has been demonstrated in the central nervous system of the miracidium of Schistosoma mansoni by using acetylthiocholine and O-acetyl-5-bromoindoxyl as substrates in association with inhibitors.

2. No histochemical evidence of any nonspecific esterases, lipases or of pseudocholinesterases could be found.
QUANTITATIVE APPRECIATION BY COMPARATIVE AUTORADIOGRAPHY OF THE SYNTHESIS AND MAINTENANCE OF SOME ORGANIC CONSTITUENTS OF THE EPIPHYSEAL PLATE OF GROWING RATS, IN RELATION WITH MAGNESIUM DEFICIENCY
Authors: BELANGER LEONARD F.
Published: April 2011

A simple and reliable method for quantitative assessment of autoradiographs by comparative histophotometry has been described. Its application to the study of tissues tagged in vivo with S35 methionine or S35 sulfate, and after Ca45 in vitro uptake, has revealed regional differences in the synthesis of proteins and sulfated mucopolysaccharides, as well as in the residual amount of cation binder present in cartilage of normal growing rats. It has also revealed differences in the behavior of all three materials between normal and magnesium deficient animals.
THE REDUCTION OF TETRAZOLIUM SALTS BY ASCITES TUMOR CELLS IN THE PRESENCE OF GLUCOSE
Authors: BARKA T. , DALLNER G.
Published: April 2011

The Ehrlich ascites tumor cells reduce tetrazolium salts intensely in the presence of glucose. The analysis of this glucose staining indicates the role of an electron-carrier in the glycolysis, probably of glycerolaldehyde-dehydrogenase. The qualitative and quantitative aspects of glucose-staining were investigated and discussed.
METACHROMASIA INHIBITING COMPONENTS IN AMYLOID
Authors: LARSEN BØRGE
Published: April 2011

I): The degree of inhibitory effect of serum proteins and a periodic acid Schiff-positive amyloid fraction on metachromasia resulting from a metachromatic amyloid fraction was studied and compared to the metachromatic potency of chondroitin sulfate exposed to the same type of inhibition. The metachromatic properties of chondroitin sulfate were close to those of the metachromatic amyloid fraction.

II): In comparing the metachromasia obtained with toluidine blue and methyl violet, it was noted that on incubation with the periodic acid Schiff-positive amyloid fraction, amyloid metachromosia after methyl violet staining was inhibited less than after toluidine blue staining.

III): It is emphasized that a competition between dye and proteins occurs in metachromatic staining reactions.

IV): Inhibition of amyloid metachromasia in histological tissue sections could not be demonstrated with the techniques used.
HISTOCHEMICAL STUDY OF THE PIGMENT FOUND IN THE EPITHELIUM OF THE KIDNEY OF THE PHILODRY AS SCHOTTII (SCHLEGEL)
Authors: DA SILVA SASSO WILSON , DE CASTRO NYLCEO MARQUES , PACHECO IRINEO PONTES
Published: April 2011

The authors studied histochemically the pigment observed within the cells of the proximal convolutions of the kidneys of the ophidian ( Philodryas schottii), known in Brazil as "parelheira".

The results demonstrate the existence of two pigments: an argentaffin, found in greater quantity and a chromolipoid of the lipofuscin type, in smaller amounts.
PRELIMINARY STUDIES ON THE HISTOLOGICAL DEMONSTRATION OF DESOXYRIBONUCLEASE II BY ADAPTATION OF THE GOMORI ACID PHOSPHATASE METHOD
Authors: ARONSON J. , HEMPELMANN L. H. , OKADA S.
Published: April 2011

An attempt has been made to adapt the acid phosphatase method of Gomori to the demonstration of desoxyribonuclease II activity. The procedure as used is subject to the disadvantages of the original method. The enzymatic activity may be demonstrable only when desoxyribonuclease and nucleotidase activity occur in the same types of cells. Although the staining occurred mainly within discrete cells, the intracellular localization apparent under these conditions of study may well be artifact. In the spleen, the stainning occurs mainly in the red pulp and in a ring of cells, possibly macrophages, surrounding the lymphatic nodules. Hepatic and Kupffer cells stain as do cells in the tubules of the kidney cortex and in the intestinal mucosa.
THE EFFECT OF PHOSPHOMOLYBDIC ACID ON THE STAINABILITY OF CONNECTIVE TISSUES BY VARIOUS DYES
Authors: PUCHTLER HOLDE , ISLER H.
Published: April 2011

The mechanism of the selective staining of connective tissue obtained in trichrome methods (e.g. Masson, Heidenhain, Goldner) by "mordanting" with phosphomolybdic acid was investigated in sections of rat intestine.

Information on the distribution of phosphomolybdic acid was obtained by treating sections with this acid and then with stannous chloride. The molybdenum blue thus obtained was located in those very structures which were stained selectively in trichrome methods.

The chemical characteristics of the dyes taken up after phosphomolybdic acid treatment were investigated by comparing sections stained with acid, basic and amphoteric dyes. In the absence of phosphomolybdic acid, only dyes with acidic groups yielded a staining of connective tissue, but this staining was not selective. In sections pretreated with phosphomolybdic acid, only dyes with basic groups stained connective tissue, and only dyes with strong basic groups yielded a selective staining of these structures. Basic and amphoteric dyes applied after phosphomolybdic acid had the same distribution as this acid.

It is concluded that the polyvalent phosphomolybdic acid is forming a bridge between the substrate and the dye, holding the dye and probably also the substrate by their cationic groups.
EFFECTS OF FIXATION IN LIPOID HISTOCHEMISTRY
Authors: ELFTMAN HERBERT
Published: April 2011

A variety of methods of lipoid fixation are needed to satisfy the requirements of different investigations. Of greatest current usefulness are the methods which involve dichromate oxidation of the tissue. Useful as the study of frozen sections of such tissue has been in the past a wider horizon of application has been opened by controlled chromation which allows lipoid histochemistry to be studied in paraffin sections as a routine procedure.
STUDIES IN HISTOCHEMISTRY. II. QUANTITATIVE HISTOCHEMICAL ESTIMATION OF ACTIVITIES OF ALKALINE AND ACID PHOSPHOMONOESTERASES
Authors: TAFT E. B. , SCOTT J. F.
Published: April 2011

Methods have been described for the quantitative histochemical estimation of alkaline and acid phosphomonoesterase activities of sections of fresh frozen tissue. Both methods employ sodium β-glycerophosphate as substrate but are not limited to this particular substrate. The methods are essentially microbiochemical and permit the accurate estimation of enzyme activity in sections adjacent to those prepared for histological examination or for the histochemical localization of sites of activity by one of the conventional qualitative technics. The activities of these enzymes in sections of normal fasted rat liver 3-4 mm. in diameter and 20 microns thick are: 5.6 ± 0.7 mµg. phosphorus/µg. protein/30 minutes at pH 9.4 and 37°C. and 29.6 ± 4.6 mµg. phosphorus/µg. protein/30 minutes at pH 5.0 and 37°C.
IMPROVED TECHNIQUE FOR THE HISTOCHEMICAL DEMONSTRATION OF SUCCINIC DEHYDROGENASE IN BRAIN TISSUE
Authors: FRIEDE REINHARD
Published: April 2011

A tissue cutting technique for the histochemical demonstration of succinic dehydrogenase activity in brain tissue is described. The demonstration of succinic dehydrogenase in fine and delicate structures depends on a tissue temperature of –3 to –2°C. during cutting. Artefacts such as coarse granules, loss of activity and diffusion of stain are prevented by minimizing of traumatic effects due to this improved technique.
STAINING PROPERTIES OF INDICATOR DYES IN NONAQUEOUS SOLUTION, IN RELATION TO SURFACE ACID AND BASIC STRENGTH OF TISSUE PROTEINS, AND TO METACHROMASIA
Authors: LANDING BENJAMIN H. , WILSON HAZEL E.
Published: April 2011

The staining properties of 69 color indicator dyes and 31 fluorescent indicators in isooctane (2,2,4-trimethylpentane) solution were studied. Thirty one of 56 indicators of pK below approximately 6 stained test tissue sections, 25 with the basic color, 2 with the acid color, and 4 with colors difficult to characterize. Conversely, only 10 of 44 indicators with pK's above 6 stained tissue sections, 5 with the acid color, 3 with the basic color, and 2 with unassignable color. This behavior pattern is independent of the chemical nature of the indicator dye, and appears to reflect a general property of the form of combination between tissue proteins and indicator dyes under the conditions used. The findings suggest that this combination has the nature of a salt-link, that staining by most indicator dyes in a nonpolar solvent is related to acceptance of hydrogen ions (protons) by the tissue proteins, that the different tissue proteins have relatively uniform proton-accepting power under the conditions used, and that they behave in general like relatively weak bases. The ability of tissue proteins to determine the staining color of certain dyes provides a possible explanation for some instances of metachromasia.
PHYSICAL AND CHEMICAL PROTECTION OF CELL CONSTITUENTS AND THE PRECISE LOCALIZATION OF ENZYMES
Authors: SCARPELLI D. G. , PEARSE A. G. E.
Published: April 2011

1. Physical and chemical protection of cytoplasmic constituents in tissue sections from the time of tissue removal until completion of the histochemical reaction resulted in a more precise intracellular localization of various enzymes.

2. In many instances sites of enzymatic activity were localized on rod-shaped and ovoid bodies resembling mitochondria and lysosomes.

3. Thermal, osmotic and chemical injury of cytoplasmic structures produced extensive intracellular and extracellular diffusion of enzyme and reaction product.

4. Once thermal damage had occurred mitochondrial swelling and enzyme diffusion appeared despite osmotic and chemical protection.
THE SITE OF A DIFFUSIBLE NUCLEOTIDE REVEALED BY A COMPARISON OF CHEMICALLY FIXED TISSUE WITH FROZEN AND DRIED TISSUE
Authors: BELL L. G. E.
Published: April 2011

1. Using unfixed frozen and dried tissue as a standard it is shown that acid alcohol fixation extracts a diffusible nucleotide from the vicinity of the chromosomes.

2. It is shown that the denuded area contains protein and that the end groups are not available for reaction with protein reagents.
EVALUATION OF INDOPHENOL DERIVATIVES AS REAGENTS FOR DEMONSTRATING CYTOCHROME OXIDASE IN TISSUE SECTIONS
Authors: CRAWFORD DAVID T. , NACHLAS MARVIN M.
Published: April 2011

Seven redox dyes were investigated as possible histochemical indicators of cytochrome oxidase activity. All failed to meet one prime requisite, namely, that the oxidized product remain at the site of enzymatic action.

The "color fading" phenomenon was studied. This effect was one in which sections and homogenates became colored as a result of oxidase activity and then faded during further incubation. Evidence was provided that this reaction occurred as a result of endogenous dehydrogenase activity, mainly succinic dehydrogenase. The decoloration in solution could be prevented by N-ethyl maleimide, without inhibiting cytochrome oxidase.
QUANTITATIVE EVALUATION OF THE ALKALINE GLYCEROPHOSPHATASE ACTIVITY OF THE LIVER DURING DEVELOPMENT AND OF THE MEDIUM USED FOR ITS CULTURE IN VITRO
Authors: ROSSI F. , BONSIGNORE A. , REALE E. , VIVORI E. , LUZZATTO L.
Published: April 2011

We have quantitatively determined the variations of the liver alkaline glycerophosphatase rate during and after ontogenesis. The glycerophosphatase rate of plasma, of embryonic extract and of the medium employed for the culture of chick organ primordia were also determined. This investigation shows an initial diminution of the enzymic rate from the 4th to the 6th incubation day and an increase after hatching. We have observed that the glycerophosphatase activity of the medium is higher than that of the sum of single activities of plasma and embryonic extract. This fact emphasizes the importance of the composition of the medium in research on alkaline glycerophosphatase activity in vitro.
HISTOCHEMICAL DEMONSTRATION OF ACID PHOSPHATASE ACTIVITY IN OSTEOCLASTS
Authors: BURSTONE M. S.
Published: April 2011

High acid phosphatase activity was observed in osteoclasts of several species using a reproducible azo-dye technique.

High activity of two distinct enzymes, acid and alkaline phosphatase, are associated with osteoclasts and osteoblasts respectivey.

Althouth frozen-dried tissues are recommended for definitive studies, the enzyme techniques used give satisfactory results with cold acetone-fixed tissues. The most accurate localization of acid phosphatase in osteoclasts in controlled comparative studies is obtained with double-embedded frozen-dried undecalcified tissues in conjunction with naphthol AS-phosphates.
A QUANTITATIVE HISTOCHEMICAL METHOD FOR THE MEASUREMENT OF DEHYDROPEPTIDASE I
Authors: ALLEN NORMAN
Published: April 2011

A method is described for the quantitative histochemical determination of dehydropeptidase I based upon the spectrophotometric procedure of Carter and Greenstein (1946). The method will measure enzyme activity in most tissues with samples of 30 to 50 µg dry weight. In a very active tissue such as kidney only 10 µg are required. Absolute ethanol has been suitable for precipitating protein and reducing the blank. Multiple determinations on the same tissue sample show good agreement, with coefficients of variation of less than 8. Optima for incubation time and tissue concentration are described.
SAPONIFICATION AS A MEANS OF SELECTIVELY REVERSING THE METHYLATION BLOCKADE OF TISSUE BASOPHILIA
Authors: SPICER S. S. , LILLIE R. D.
Published: April 2011

Saponification reverses the methylation blockade of cartilage and mast cell basophilia demonstrated by azure A above pH 3-4, but fails to do so at pH levels below this range. Saponification of oxidized hair cortex restores the methylation blocked basophilia demonstrable at pH 2 or higher. Mouse amyloid is characterized by a weak basophilia which is relatively resistant to methylation.

The results are interpreted as indicating that methylation hydrolyzes the sulfate esters and esterifies other acid groupings in tissue sections.
A METHOD FOR THE CYTOCHEMICAL DEMONSTRATION OF SUCCINIC DEHYDROGENASE IN HUMAN LEUCOCYTES
Authors: DE SOUZA E. J. , KOTHARE S. N.
Published: April 2011

1. A method for demonstrating succinic dehydrogenase in human leucocytes using a p-nitrophenyl substituted ditetrazole is described.

2. In some of the leucocytes the formazan appeared as fine blue discrete granules or rods.
EFFECTS OF ALLOXAN AND REDUCED FOOD INTAKE ON LIVER DEOXYRIBOSE NUCLEIC ACID OF THE RAT
Authors: DI STEFANO HENRY S. , DIERMEIER HAROLD F.
Published: April 2011

Effects of alloxan, reduced food intake, starvation and biopsy procedure on the deoxyribose nucleic acid (DNA) content of rat liver tetraploid nuclei have been investigated. It was found that: (1) Alloxan treatment resulted in an increased amount of DNA. (2) Reduced food intake of treated animals Played no part in this increase. (3) Complete starvation for a period of six days did not effect the amount of DNA in these nuclei. (4) An increased amount of DNA was found in tetraploid nuclei of rat liver 13 days after removal of a small biopsy sample.

The increased DNA in tetraploid liver nuclei of alloxan diabetic rats may be explained on the basis of regeneration of liver tissue due to some submicroscopic liver damage caused by the drug.
HISTOCHEMICAL INVESTIGATIONS OF A SILVER METHOD FOR AXONS
Authors: WINKELMANN R. K. , SCHMIT ROBERT W.
Published: April 2011

Silver ion is not specific either in location or in action in the present silver method. Studies with radioactive silver, with the dark-field microscope, and with simple reducing solutions demonstrate a general distribution of silver ion in the impregnated tissue. Copper and mercury may be substituted for silver in the impregnating solution. A water-soluble aldehyde is necessary as a fixative. The substrate for staining appears to require the presence of protein-bound amine groups. The product-forming solution does not require the presence of a reducing agent. Sulfite ion is not necessary. All controllable sources of base that were utilized produced the histologic results. The product appears to be silver oxide in terms of the present solubility studies.
A SCHIFF REAGENT OF CALIBRATED SENSITIVITY
Authors: ELFTMAN HERBERT
Published: April 2011

The sensitivity of Schiff reagent for the detection of low concentrations of aldehyde increases progressively as the concentration of sulfurous acid in the reagent is lowered. Calibration of sensitivity is readily accomplished by titrating the Schiff reagent against the iodine of Lugol's solution.

Stability of Schiff's solution depends on decreasing the evaporation of sulfur dioxide and of retarding the change of sulfite to sulfate. For practical purposes these ends can be achieved by storage in the refrigerator of a solution with greater sulfurous acid content and acidity than the solution used for staining. Evacuation of the excess sulfur dioxide or mixing in proper proportions with used reagent allows the rapid production of a solution of the desired sensitivity. If chemical preservatives are required, 0.5% hydroquinone gives very good results.

Spectrophotometric measurement of the colored solutions produced by adding aldehyde to Schiff reagent indicates that at least two distinct compounds may result. The one that is usually mentioned as characteristic of a positive response of aldehyde to the Schiff reagent has a violet color. When the concentration of aldehyde is so low that only a Schiff solution of low sulfurous acid titer will react with it, a red color is produced. Spectrophotometric analysis shows this compound to be different from the original basic fuchsin and validates its use as an endpoint in the reaction of aldehyde with Schiff's reagent.
CHOLINESTERASES IN THE MAMMALIAN TONGUE
Authors: ELLIS RICHARD A.
Published: April 2011

Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) have been demonstrated histochemically in the tongues of five Primates and the rabbit. Sensory nerve endings in the tongue, including the subepithelial nerve net, Meissner's corpuscles and the end bulbs of Kraus, contain both AChE and BChE. The intraepithelial nerve endings and the taste buds in the foliate, vallate and fungiform papillae react for AChE only. The postganglionic parasympathetic nerve fibers innervating both the pure mucous and the pure serous glands of the tongue have abundant AChE. In addition to the AChE and BChE activity in their motor end plates, the skeletal muscle fibers of the tongue also have a concentration of cholinesterases at their point of insertion on the lingual fascia which is designated as the cholinesterasic apparatus.

The Winkelmann and Schmit silver technique for nerve axoplasm is found to be unreliable for visualizing autonomic nerves.
ENZYMATIC HISTOCHEMISTRY OF ETHIONINE INDUCED LIVER CIRRHOSIS AND HEPATOMA
Authors: WACHSTEIN MAX , MEISEL ELIZABETH
Published: April 2011

A number of enzymatic histochemical staining reactions were applied to the livers of rats that had been given dl-ethionine for periods up to 330 days. The ethionine lesion is characterized by early central hepatocellular necrosis, proliferation of oval cells, and degenerative and regenerative alterations in the liver cells leading to cirrhosis and areas of adenofibrosis. Finally, liver cell carcinoma develops.

The zonal differences of various enzymatic staining reactions in the liver lobules characteristic for the normal liver disappear in regenerating liver lobules. Most enzymes are demonstrable in these regenerating cells, as for instance succinic dehydrogenase, nonspecific esterase, glucose-6-phosphatase, diphosphopyridine nucleotide diaphorase and others. Hepatic carcinoma cells likewise retain many of the staining reactions. There is, however, a complete lack of bile canalicular activity, which is best seen with the ATPase technique. Such a deficiency occurs also in some regenerating nodules without obvious evidence of malignancy. Thus, the lack of bile canalicular staining in cirrhotic liver lobules may indicate impending carcinomatous transformation.

Oval "ductular" cells showed a lack of various enzymatic staining reactions, but did show moderate DPN diaphorase and nonspecific esterase activity. The origin of these cells from liver cells, although not proven, is definitely suggested. In areas of oval cell proliferation, there was marked increase in the intensity of sinusoidal staining with various phosphatase techniques. Fibrous tissue, particularly in areas of adenofibrosis, showed areas of marked adenosine triphosphatase activity. On the other hand, 5-nucleotidase activity was increased strongly in areas where reticulum fibers had been deposited. Increase in nonspecific esterase and acid phosphatase in Kupffer cells indicated apparently a marked stimulation of the reticuloendothelial system.
RADIOAUTOGRAPHIC STUDIES ON NUCLEAR ACTIVITY IN THE LIVER
Authors: FICQ A.
Published: April 2011

The present paper deals with observations of nucleic acid and protein metabolism (uptake of radioactive precursors) in liver nuclei either in vivo or in isolation. In the latter case cytoplasmic fractions appear to enhance the metabolism. Metabolism is also increased following partial hepatectomy. The liver has further been envisaged as playing a role in immunological reactions and the fixation of antigens has been demonstrated both in Kupffer and parenchymal cells.
CYTOPLASMIC PARTICLES AND SERUM ALBUMIN SYNTHESIS
Authors: PETERS THEODORE
Published: April 2011

The formation of serum albumin in chicken liver has been shown to proceed via an intermediate stage in which newly formed albumin is bound to microsomes. This bound albumin is apparently formed within 2-3 minutes, but its release requires 15-20 minutes and is readily blocked by metabolic inhibitors. Comparison of the effectiveness of various agents in releasing this newly formed albumin suggests that the albumin is bound on or in the lipide-rich membrane of the microsome. After release by desoxycholate the material is indistinguishable from native serum albumin in its physical and chemical properties. The mechanisms of the synthesis of proteins in liver are discussed, and insofar as possible related to the results with serum albumin.