bluuebunny/crossref_metadata_2025_split huggingface dataset:
CYTOCHEMICAL LOCALIZATION OF NUCLEIC ACIDS BY AN ACRIFLAVINE-PHOSPHOTUNGSTATE COMPLEX FOR FLUORESCENCE MICROSCOPY AND ELECTRON MICROSCOPY
Authors: CHAN-CURTIS VICTORIA , BELT W. DUANE , LADOULIS CHARLES T. 
Published: April 2011

An acriflavine-phosphotungstate complex (A-PTA) was synthesized from a reaction between acriflavine and phosphotungstic acid, with the former in excess. The purified product is fluorescent and electron-dense, and was studied for its use as a cytochemical reagent for fluorescence microscopy and electron microscopy. A selectivity for nucleic acids and sulfated cerebroside esters that excludes sulfated glycoaminoglycans is proposed. The reaction for fluorescence microscopy is rapid, sensitive and reproducible. The electron-dense A-PTA complex localized nucleic acids for electron microscopy. Known sites of DNA and RNA in cells of pancreatic, cerebellar and testicular tissue have been demonstrated with the reaction. The reaction is administered as a single step, interposed in the electron microscopic preparative procedure, and can be used in either an alcoholic or an aqueous system. The alcoholic A-PTA reaction produced adequate contrast on short duration (2 hr); the aqueous block reaction was found most satisfactory when prolonged to 48 hr. Uranyl acetate, phosphomolybdic acid and phosphotungstic acid can be used as supplementary stains in addition to the A-PTA reaction. Under these conditions, DNP and RNP sites were demonstrated. The A-PTA complex is a prototype of the coupled dye-metal histochemical reagents, and its selectivity for nucleic acids represents a new approach in cytochemistry. Its reactions can be investigated by fluorescence microscopy and electron microscopy, and in solution. Other dye-metal reagents can probably be formulated on a similar design as detection systems for lipids, polysaccharide or specific groups in proteins.
CONTROLLED CHROMATION
Authors: ELFTMAN HERBERT 
Published: April 2011

Controlled chromation represents another step in the development of dichromate oxidation as a cytochemical technique, to which previous contributions have been made by Weigert, Smith, Dietrich, Ciaccio and Baker. Recognition of the critical importance of controlling pH, in addition to the regulation of temperature, time, and concentration, allows voluntary adjustment of these variables to fit each particular problem. Control of chromation is equally important whether it is exercised on fresh tissues, frozen sections, or tissues previously subjected to the action of fixatives or solvents. By using graded intensities of chromation, differential visualization of a series of cell constituents may be achieved. Judicious combination of controlled chromation with the use of other reagents, including solvents, offers the possibility of developing analytical methods for the study of a great variety of lipid and non-lipid tissues components. The relative ease with which phospholipids, such as those of the Golgi apparatus, are chromated allows their routine visualization to be accomplished after chromation at pH 3.5 for 18 hours at 56°C. with 2.5% potassium dichromate, followed by paraffin embedding. Alternate sections stained with Sudan black B and with hematoxylin can be ready for study within twenty four hours of the collection of the tissue. Chromated lipids are stained by both Sudan black B and by hematoxylin, non-chromated lipids only by Sudan black B, and non-lipid chromated substances only by hematoxylin. Other dyes may be used in place of Sudan black B and hematoxylin when advantageous. The staining of chromated tissue with hematoxylin can be controlled by the addition of any one of several reagents to the staining solution, so as to restrain the formation of hematoxylin lakes by minor concentrations of chromium. Differentiation is consequently accomplished during staining and the entire process can be completed on paraffin sections at 56°C. in two hours.
ATTEMPTS TO DEMONSTRATE ACETYLCHOLINESTERASE ACTIVITY IN BLOOD AND BONE-MARROW CELLS BY A MODIFIED THIOCHOLINE TECHNIQUE
Authors: ZAJICEK J. , SYLVEN B. , DATTA N. 
Published: April 2011

The thiocholine method of Koelle (1951) was used in attempts to demonstrate acetylcholinesterase activity in artificial enzyme-gelatin mixtures and in suspensions and smears of isolated platelet, ghosts, erythrocytes, and megakaryocytes of different species. When the last processing step with ammonium sulphide was omitted, the following observations were made: After a short time of incubation, small granular or rod-like deposits were found at the presumed sites of AChE activity. These deposits apparently grew into large crystals during prolonged incubation. Such crystals were demonstrated at the surface of platelet ghosts and erythrocytes of species bearing AChE activity, and further in megakaryocytes of some species presumably bearing this activity. The crystals were provisionally assumed to be a Cu thiocholine salt representing sites of AChE activity. If an enzyme inhibitor was added or a substrate used which is not split by AChE no crystals appeared.
THE HISTOCHEMICAL DISTRIBUTION OF 5-NUCLEOTIDASE AND UNSPECIFIC ALKALINE PHOSPHATASE IN THE TESTICLE OF VARIOUS SPECIES AND IN TWO HUMAN SEMINOMAS
Authors: WACHSTEIN M. , MEISEL E. 
Published: April 2011

Sections of human adult testicles and of testicles from dog, rabbit, guinea pig, rat and mouse as well as of two seminomas were stained for nonspecific alkaline phosphatase and for 5-nucleotidase. The most outstanding feature in the testicles of all species examined is the presence of 5-nucleotidase predominantly located in the nuclei of the spermatogenic elements. In the two seminomas 5-nucleotidase was likewise found in the nuclei of the tumor cells. The validity of these findings is discussed and it is concluded that the predominantly nuclear localization reflects the true distribution of 5-nucleotidase in these cells. The seminomas are not only rich in 5-nucleotidase, but contain apparently considerably larger amounts of nonspecific alkaline phosphatase than do normal human spermatogenic cells. The seminomas, therefore, show a distinct staining pattern as regards 5-nucleotidase and alkaline phosphatase so far apparently not observed in other tumors.
THE EFFECT OF METHYLATION ON BASOPHILIA
Authors: FISHER EDWIN R. , LILLIE R. D. 
Published: April 2011

A mixture of 0.05 N HCl and methanol is capable of completely inhibiting thionin and azure A metachromasia of intestinal mucin, cartilage and mast cell granules after treatment for 2 hours at 58°C. or 48 hours at room temperature. Slight reduction is noted after 2 weeks at 3°C. Cytoplasmic basophilia is completely destroyed after 4 hours of methylation at 58°C. or 48 hours at room temperature. Pancreatic acinar cell cytoplasm appears more resistant, requiring 8 hours at 58°C. and longer than 9 days at room temperature. Methylation at 3°C. is ineffective in inhibiting this basophilia even after 2 weeks. Nuclear and nucleolar basophilia with aniline dyes (azure A and thionin) requires 24 hours of methylation at 58°C. or 14 days at room temperature before there is complete destruction. There is no effect produced by methylation at 3°C. for 2 weeks. Alum hematoxylin staining of these structures is unaffected by the methylation procedure, indicating the different nature of action of this dye complex from the aniline dyes used. Reversal of the methylation procedure or demethylation can be accomplished with 0.5% KMnO4 for ½ hour at room temperature provided the methylation has not been too prolonged. Other oxidants such as periodic, peracetic and chromic acids as well as aqueous HCl and alkali (borax solution) fail to reverse the blockade. The substitution of 0.1 N HCl for the usual 0.05 N HCl in the methylation procedure brings about effective blockade in approximately ½ the time required by the 0.05 H NCl methanol. Despite the removal of cytoplasmic and nuclear basophilia by this method sections treated concomitantly and stained with the Feulgen nucleal procedure demonstrate strong Feulgen reactions indicating selectivity for the acid portion of the nucleic acid involved. Methylation after fixation of tissue in a chromate containing fixative (Zenker acetic) requires more prolonged treatment to inhibit cytoplasmic basophilia than is required after Formalin or 95% alcohol fixation. Otherwise the reactions occur similarly after either of the three fixatives. The positive periodic acid-Schiff reaction of colonic mucin, mixed tumor stroma and cartilage is completely inhibited by prior methylation for 96 hours at 58°C. and the reaction of gastric and small intestine mucins is only moderately reduced at this period. At room temperature no alteration is observed after methylation for 21 days. The positive reaction of human mast cell granules is abolished after 72 hours of treatment at 58°C. and is only moderately affected after treatment for 21 days at room temperature.
HISTOCHEMICAL DEMONSTRATION OF ASCORBIC ACID IN THE ADRENAL GLAND WITH A CRITICAL STUDY OF THE METHOD
Authors: ERÄNKÖ OLAVI 
Published: April 2011

The silver nitrate technique for the histochemical demonstration of ascorbic acid has been applied to the adrenals of the rat. Whole glands, fresh-frozen and frozen-dried sections were treated with the silver reagent. Diffusibility of ascorbic acid in the adrenal tissue was studied in tissue disc preparations and with "section chromatography". The following conclusions were derived: 1) Concentrations of silver nitrate, acetic acid and alcohol in the reagent are not critical. 2) Light does not affect the results under the conditions of the present study. 3) Excess silver nitrate must be washed out before fixation in thiosulphate. 4) Ascorbic acid is in a highly diffusible form in the adrenal cortex. 5) The solvent penetrates adrenal tissue at a higher speed than silver nitrate. 6) The presence of silver granules in the zona glomerulosa and the medulla of glands immersed as a whole into the silver reagent is due to dislocation of ascorbic acid during the histochemical procedure. 7) Diffusion of ascorbic acid is reduced to the minimum by treating fresh-frozen or frozen-dried sections with the silver reagent. Silver granules are in such sections restricted to the fasciculata and reticularis. 8) Stimulation of the adrenal cortex with injections of adrenaline or ACTH cause a depletion of silver granules mainly in the lower parts of the fasciculata and the reticularis. 9) Histochemically demonstrable ascorbic acid is probably distributed evenly in the cell both in normal and in stimulated adrenals. This point requires further study.
SULPHATION TECHNIQUES IN HISTOCHEMISTRY WITH SPECIAL REFERENCE TO METACHROMASIA
Authors: KRAMER H. , WINDRUM G. M. 
Published: April 2011

1. Methods are described for the sulphation of tissue carbohydrates in histological sections. 2. Evidence is produced to show that the various procedures described result in the formation of sulphuric acid esters of the carbohydrates. 3. The histochemical applications of the metachromatic staining reaction can be greatly extended by the use of these techniques, as neutral polysaccharides, non-sulphated acid mucopolysaccharides, and glycoproteins, which are ordinarily orthochromatic, are rendered metachromatic.
CYTOLOGICAL OBSERVATIONS OF THE EFFECT OF ENZYMES ON ACID-TREATED SPORES OF BACILLUS CEREUS
Authors: KEIGLER NORMA M. , SMITH ANDREW G. 
Published: April 2011

Resting spores of Bacillus cereus were studied cytologically by means of a technique involving both acid and enzyme hydrolysis which was supplemented by selective staining and observation by both bright field and phase contrast microscopy. Enzymes were used singly and in two-enzyme sequences following acid hydrolysis. The results show the artifactitious peripheral structure to be digested by the proteases pepsin and trypsin, and the central structure to be relatively resistant to such action. The nucleases ribonuclease and desoxyribonuclease caused no dissolution of structures, but effected to different degrees a loss of basophilia in the peripheral structure. The former's lesser action led to the conclusion that some ribonucleic acid was associated with the spore's nuclear chromatin in a relatively acid resistant complex, thus, aligning the spore's nucleus with that of other plant and animal tissues in regard to the presence of a ribodesoxyribonucleoprotein complex. Other than effecting the phase contrast phenomenon of reversal of contrast, two-enzyme sequences failed to result in striking increases in degradation and dissolution of the central and the peripheral structures. Buffer and diluent effects on a substrate's subsequent susceptibility to enzyme action and to staining reactions were noted and were in accord with the finding-of other investigators. Osmium tetroxide fixation appeared to increase the subt strate's susceptibility to trypsin action. Magnesium sulfate was shown to inhibis pepsin activity. Potassium permanganate was shown to be an effective mordant for increasing the basophilia of the central structure.
THE DESOXYRIBOSE NUCLEIC ACID (DNA) CONTENT OF HUMAN MONOCYTES AND THEIR DERIVATIVES DURING GIANT CELL FORMATION IN VITRO
Authors: GOLDSTEIN MILTON N. 
Published: April 2011

1. The desoxyribose nucleic acid (DNA) content of individual cell nuclei in monocytes and their derivatives, multinucleated giant cells, has been determined by microspectrophotometry. These measurements were performed on Feulgen stained material. 2. Although most of the measurements of individual nuclei in cultures incubated from 4 to 138 hours were close to diploid value, there were variations in some nuclei well above and below this value. 3. The data indicated that DNA was being synthesized during cultivation. 4. Amitosis occurred in some of the nuclei of the multinucleated giant cells after these cells had formed by cell fusion.
ELECTRON MICROSCOPY OF BASOPHILIC COMPONENTS OF CYTOPLASM
Authors: PORTER KEITH R. 
Published: April 2011

Electron microscopy of thinly spread cells grown in vitro has defined, as a component of the cytoplasm, a complex reticulum of strands and vesicles which has come to be called the endoplasmic reticulum. The component is limited by a membrane similar in thickness (ca. 80 Å) to the plasma membrane of the cell. It separates the content of the strands and vesicles from the general matrix of the cytoplasm and gives to the whole component the character of a finely divided vacuolar system. The elements of the system are frequently large enough to be resolved by light microscopy and can be shown to have an affinity for basic dyes. The system has therefore been identified as a basophilic component of the cytoplasm (25). With the techniques now available for thin sectioning the same system of vesicles and interconnecting canaliculi can be demonstrated in cells fixed in situ and studies of such material have defined the system as a universally occurring component of the cytoplasm. In its structure it varies greatly from one cell type to another and even within a single kind of cell under different physiological states. In one form it is common for it to be made up of extraordinarily thin, flattened vesicles referred to as cisternae. In certain types of cells, particularly acinar cells of exocrine glands, these vesicles are organized in parallel arrays and such organizations have been shown to coincide with the distribution of the basophilic component.. Thus in thin sections as in cultured cells the reticulum is identified with the basophilic material. In very recent electron microscopy of thin sections, Palade (16) has observed that a dense granular component of the cytoplasmic matrix is usually adjacent to the membrane surfaces of the reticulum. In rapidly growing cells where the basophilia is diffuse and where elements of the reticulum may not be prominent, granules of this latter character (100-300 Å diameter) are abundant and scattered throughout the cytoplasmic matrix. This suggests therefore that in the other instances where the intensely basophilic component and the endoplasmic reticulum coincide, the latter structure owes its basophile properties to the quantities of the granular material which associate preferentially with its membrane surfaces. This conclusion finds support in studies of cell fractions (1, 23) which have defined small cytoplasmic particles that are extraordinarily rich in ribonucleic acid. It is also supported by the observation that parts of the nucleolus consist of closely packed granules of similar size and density. It appears then that the basophilic component when it exists as a distinct and localized entity of the cytoplasm, consists of a concentration of vesicular and canalicular elements of the endoplasmic reticulum with small basophilic granules associated. Where the basophilia is diffuse it seems to be an expression of the diffuse distribution of the small granules.
THE EFFECTS OF RIBONUCLEASE ON THE RIBONUCLEIC ACID AND ENZYME ACTIVITIES OF MICROSOMES ISOLATED FROM RAT LIVER HOMOGENATES
Authors: NOVIKOFF ALEX B. , RYAN JEAN , PODBER ESTELLE 
Published: April 2011

1. The RNA of isolated rat liver microsomes is readily accessible to ribonuclease digestion. About 85% of the RNA-ribose is removed by the nuclease; data in the literature suggest that the rest remains in smaller polynucleotides rather than RNA per se. 2. Digestion of microsomes with ribonuclease dissolved in glycyl-glycine buffer, pH 7.0, has no effect upon the activities of the enzymes studied: glucose-6-phosphatase, esterase, triacetic acid lactonase, adenosine-5'-phosphatase, acid phosphatase, adenosine triphosphatase, and succinoxidase. 3. Digestion with ribonuclease dissolved in water has no effect upon the acid phosphatase and adenosine-5'-phosphatase activities, while esterase, adenosine triphosphatase and succinoxidase activities are somewhat lower. Glucose-6-phosphatase activity is markedly reduced, as it is by incubation of microsomes with either water or buffer alone.
HISTOCHEMICAL AZO COUPLING REACTIONS A CATECHOLAMINE IN ENTEROCHROMAFFIN CELLS IN PLACE OF OR IN ADDITION TO 5-HYDROXYTRYPTAMINE
Authors: LILLIE R. D. , PIZZOLATO P. , VACCA L. L. , CATALANO R. A. , DONALDSON P. T. 
Published: April 2011

Appreciable azo coupling of enterochromaffin cells (EC) and of adrenal medulla (AM) is restricted to the alkaline range, fading out around pH 6, negative at 3-5. Low pH (3.0) azo coupling of rat and mouse mast cells (MC) with diazosafranin correlates with their known content of 5-hydroxytryptamine (5-HT) and with the pH 3-9 range of deep red azo coupling color reaction of p-nitrodiazobenzene with 5-hydroxytryptophan and 5-HT. The MC reaction is covalent azo bonding and not cationic dye salt bonding. The MC reaction is not influenced by prior oxidation by HIO4, FeCl3, K2Cr2O7 or I2/CH3OH exposures which prevent azo coupling of EC and of AM. The oxidation blockades of the azo coupling of EC and AM are reversed by Na2S2O5, N2S2O4 or Na2S2O3 reductions. These reductions after oxidation do not affect the MC diazosafranin reaction. In vitro the azo coupling of 5-HT is only slightly retarded but not weakened by HIO4 oxidation and then slightly enhanced by Na2S2O3, while noradrenaline gives red before oxidation, light yellow after, and after Na2S2O3, again deep red, the control α-naphthol giving deep red with p-nitrodiazobenzine at all three phases, before HIO4, between HIO4 and after Na2S2O3. Indole reactions for EC have been reported on glutaraldehyde (G), acetaldehyde and acrolein fixed but not formaldehyde (F) fixed guinea pig duodenum by Solcia and Sampietro (63), Geyer (27) (G), Barter and Pearse (5, 6) and Lillie and Greco-Henson (44). Repetition of the Lillie and Greco-Henson test of combining in the same preparation a blue indole reaction (postcoupled benzylidene) and a red azo coupling reaction with p-nitrodiazobenzene in both sequences disclosed numerous red EC and blue Paneth cells and eosinophil leukocytes, alike on F and G tissue. Direct observation during the second reaction and color photography of the same field after each of the two reactions showed that no red stained cell was replaced by a blue or purple one, and that no blue stained cell altered its color to purple or red. Moreover, azo positive cells were seen in the epithelium of the tips of the villi and in the sides of the gastric glands of the guinea pig pylorus and fundus. Indole positive cells did not occur in villus tip or pyloric gland epithelium and were restricted to fundus chief cells. It is concluded that the azo reactive substance in EC is a catechol and not 5-HT. The fluorescence studies have indicated the presence of 5-HT in EC, but the quantity present is insufficient for the acid azo coupling reaction shown by rat and mouse MC with diazosafranin.
AN EVALUATION OF HISTOCHEMICAL METHODS FOR PROTEIN GROUPS
Authors: BURSTONE M. S. 
Published: April 2011

The dinitrofluorobenzene, the ninhydrin-Schiff, alloxan-Schiff, and chloramine-Schiff methods are useful for the demonstration of certain protein groups in tissues. The coupled tetrazonium and nitrobenzoylation methods present certain difficulties in interpretation. However, the latter may be of value as a collagen or elastic tissue stain.
FLAME PHOTOMETRIC DETERMINATION OF POTASSIUM IN MICROGRAM QUANTITIES OF TISSUE AND THE DISTRIBUTION OF POTASSIUM AND LIPID IN THE ADRENAL OF THE MONKEY AND GUINEA PIG (STUDIES IN HISTOCHEMISTRY XXXII)
Authors: GLICK DAVID , SWIGART RICHARD H. , NAYYAR SOM N. , STECKLEIN HELEN R. 
Published: April 2011

A flame photometric method for the determination of potassium in microgram quantities of tissue was described. A simple water extraction of the potassium from microtome sections of adrenal tissue was shown to be preferable to either digestion or dry-ashing in the preparation of the sample for analysis. The quantitative histological distributions of total dry matter, total lipid, and potassium in the adrenal glands of M. rhesus and cynomolgus monkeys and guinea pigs were determined. No significant strain or species differences were observed among the data collected. The total lipid in the fascicular zone reached maximum concentrations of 55-70% of the total dry-weight. The potassium concentration remained within the approximate range of 1.4-1.7% of the fat-free dry-weight throughout all of the regions of the adrenals of the animals used.