bluuebunny/crossref_metadata_2025 huggingface dataset:
A SIMPLIFIED METHOD FOR THE HISTOCHEMICAL LOCALIZATION OF CARDIAC CATECHOLAMINE-CONTAINING NERVE FIBERS
Authors: LATIES A. M. , LUND R. , JACOBOWITZ D.
Published: April 2011

An improved, simplified method for the demonstration of the adrenergic innervation to the heart based on perfusion fixation is described. After perfusion with phosphate-buffered paraformaldehyde, specimens of heart tissue are frozen on Dry Ice, cut in the cryostat at –10°C and exposed to formaldehyde gas at 80°C for 30 min. The perfusion solution limits diffusion and makes it possible to show catecholamine fluorescence in cryostat sections of the heart of a quality heretofore only possible in freeze-dried tissue. The elimination of the need for freeze-drying allows studies of the effects of drugs or other procedures to be carried out in a single day with relatively simple equipment. Fixation by the perfusion solution used has been shown to be compatible with modern neurohistologic methods and to be of good quality for studies of ultrastructure. When perfused tissues are freeze-dried, catecholamine fluorescence of high quality, fully comparable to that possible with nonperfused tissue, is found. Freeze-drying of perfused heart eliminates the necessity and value of exposure to paraformaldehyde gas.
THE DEMONSTRATION OF ACID HYDROLASE ACTIVITIES IN THE INCLUSION BODIES OF TYPE II ALVEOLAR CELLS AND OTHER LYSOSOMES IN THE RABBIT LUNG
Authors: GOLDFISCHER SIDNEY , KIKKAWA YUTAKA , HOFFMAN LEE
Published: April 2011

Type II alveolar epithelial cells show high levels of acid phosphatase, aryl sulfatase B and β-glucuronidase activities. In formalin- and glutaraldehyde-fixed rabbit lung acid phosphatase and aryl sulfatase B activities were demonstrated in the cytoplasmic inclusion bodies, supporting their identification as lysosomes. Type II cells differ from alveolar macrophages in their levels of hydrolase activity and the fine structure of their active sites.
A METHOD FOR DIFFERENTIATING DOPAMINE FROM NORADRENALINE IN TISSUE SECTIONS BY MICROSPECTROFLUOROMETRY
Authors: BJÖRKLUND ANDERS , EHINGER BERNDT , FALCK BENGT
Published: April 2011

Fluorophores induced from noradrenaline and dopamine in tissue sections by treatment with formaldehyde can be separated by their different behavior upon HCl treatment. The noradrenaline fluorophore converts to a fully aromatic isoquinoline while the dopamine fluorophore remains in the form of a nonquinoidal dihydroisoquinoline. The two fluorophores have readly distinguishable excitation spectra. The procedure has been tested both in model systems and in tissue sections. The microspectrofluorometer used is described.
NUCLEOSIDE PHOSPHATASE ACTIVITIES AND SUBMICROSCOPIC LOCALIZATION OF ADENOSINE TRIPHOSPHATASE ACTIVITY IN GUSTATORY EPITHELIUM
Authors: IWAYAMA TAKASHI
Published: April 2011

Phosphatase activities were histochemically examined in the gustatory region, using adenosine triphosphate (ATP), inosine triphosphate, ADP, AMP and Na-β-glycerophosphate as substrate, and p-chloromercuribenzoate as the inhibitor for ATPase. With ATP as substrate, the reaction product was observed on taste hairs, the boundary of taste bud cells, subgemmal nerve fibers and capillary walls. The ATPase activity on the boundary of taste bud cells was differentiated from the other reactive sites and was proved to be substrate-specific and sulfhydryl-dependent. With an electron microscope, the reaction product of ATPase was localized mainly in the space between the membranes of taste bud cells and on the outer leaflets. The reaction product was also observed on intragemmal nerve fibers. The activity was present on the surface of the nerve fiber facing the dark cell, whereas it was almost absent on the site in contact with the light cell. It is suggested that dark cells are supporting elements and light ones are neuroepithelial elements.
THE PARTICIPATION OF NUCLEOTIDE IN THE FORMATION OF PHOSPHATASE REACTION PRODUCT: A CHEMICAL AND ELECTRON MICROSCOPE AUTORADIOGRAPHIC STUDY
Authors: ROSENTHAL ALAN S. , MOSES HAROLD L. , GANOTE CHARLES E. , TICE LOIS
Published: April 2011

Data are presented which show that the reaction product formed in adenosine triphosphatase cytochemical procedures is not simple lead phosphate. Both by chemical and electron microscope autoradiographic techniques, reaction product was found to contain nucleotide as well as lead and phosphate. Nearly equimolar quantities of nucleotide and phosphate are present in tissue incubated in a standard Wachstein-Meisel medium. Electron microscope autoradiographs demonstrate that nucleotide is uniformly associated with adenosine triphosphatase reaction product, regardless of its cellular distribution. In vitro studies of the interaction of nucleotide with lead and phosphate are also presented. The relationship of these observations to the validity of lead-salt cytochemical procedures is discussed.
MOUSE ERYTHROCYTE ESTERASES AND AN INTERACTION WITH HEPARIN AS SHOWN BY STARCH GEL ELECTROPHORESIS
Authors: TEMPLETON McCORMICK
Published: April 2011

Esterases from the blood of young adult male C-57 brown mice were separated by starch gel electrophoresis. Enzymatic hydrolysis of the substrates α-naphthyl acetate, naphthyl-AS acetate, α-naphthyl butyrate and acetylthiocholine was studied. Three different esterases were found in erythrocytes. One band hydrolyzed a-naphthyl butyrate but was only slightly active with acetate esters. The fastest migrating bands hydrolyzed acetate faster than butyrate esters, but would hydrolyze both substrates. A third esterase represented in the zymogram by a pair of bands (called "acetate" bands) would hydrolyze only acetate esters. When heparin, or plasma containing heparin, was inserted in a gel so as to overlap an erythrocyte sample, only the acetate bands were altered. When hepanin was inserted cathodal to an erythrocyte sample, the acetate bands are selectively affected by the heparin in such a way as to cause the acetate band to migrate more rapidly toward the anode; other esterase bands were not similarly affected. Acetate bands were not inhibited by physostygmine (1 mM) but their activities diminished with increased concentrations of buffer.
THE CYTOCHEMISTRY AND ULTRASTRUCTURE OF POLYPEPTIDE HORMONE-PRODUCING CELLS OF THE APUD SERIES AND THE EMBRYOLOGIC, PHYSIOLOGIC AND PATHOLOGIC IMPLICATIONS OF THE CONCEPT
Authors: PEARSE A. G. E.
Published: April 2011

A group of apparently unrelated endocrine cells, some in endocrine glands, others in nonendocrine tissues, share a number of cytochemical and ultrastructural characteristics. These characteristics, from the initial letters of four of which the term APUD is derived, indicate the possession of a common metabolic pattern and common synthetic, storage and secretion mechanisms. It is postulated that the various characteristics reflect the production and storage of hormone precursor protein which has a predominantly random coil conformation. Several explanations are possible to account for the APUD characteristics but if the cells, in fact, share a common ancestor the only possible candidate is the neural crest cell.
HISTOCHEMICAL DUALITY OF RABBIT INTRAFUSAL FIBERS
Authors: SPIRO ALFRED J. , BEILIN REBECCA L.
Published: April 2011

Using the adenosine triphosphatase reaction two histochemically distinct types of intrafusal fibers were demonstrated in rabbit muscle spindles despite morphologic homogeneity of these fibers. This finding, reflecting metabolic differences between these fiber types, is offered as an explanation for the previously unexplained heterogeneity of their physiologic responses.
Medical and pharmacological direct costs of a 9-week smoking cessation programme
Authors: Wasserfallen Jean-Blaise , Digon Patricia , Cornuz Jacques
Published: March 2011

Background: Medical and pharmacological direct costs of cigarette smoking cessation programmes are not covered by health insurance in several countries despite documented cost-effectiveness.

Design: prospective cost identification study of a 9-week programme in Switzerland.

Methods: A total of 481 smokers were followed-up for 9 weeks. Socio-demographic characteristics, number of outpatient visits, dosage and frequency of use of nicotine replacement therapy (NRT) as well as date of relapse were prospectively collected. Individual cost of care until relapse or programme end as well as cost per week of follow-up were computed. Comparisons were carried out between the groups with or without relapse at the end of the programme.

Results: Of the 209 men and 272 women included, 347 patients (72%) finished the programme. Among them, 240 patients (70%) succeeded in quitting and 107 patients (30%) relapsed. As compared with the group relapsing by the end of the programme, the group succeeding in quitting was more often living in a couple (68% vs. 55%, p = 0.029). Their mean weekly costs of visits were higher (CHF 81.2 ± 6.1 vs. 78.4 ± 7.6, p = 0.001), while their mean weekly costs for NRT were similar (CHF 24.2 ± 12.6 vs. 25.4 ± 15.9, p = 0.711). Mean total costs per week were similar (CHF 105.4 ± 15.4 vs. 103.8 ± 19.4, p = 0.252). More intensive NRT at week 4 increased the probability not to relapse at the end of the programme.

Conclusions: Over 9 weeks, medical and pharmacological costs of stopping smoking are low. Good medical and social support as well as adequate NRT seem to play a role in successful quitting.
Gender differences in the extent and timing of the reflected arterial waveform in the presence of coronary artery disease
Authors: Crilly Michael A , Williams David J , Clark Hazel J , Hillis Graham S , Lee Amanda J
Published: April 2011

Background: The non-invasive assessment of arterial dysfunction may improve cardiovascular (CV) risk assessment. We studied the relationship of the reflected wave transit time (RWT) and augmentation index (AIX), both derived from pulse wave analysis (PWA), in the presence/absence of coronary artery disease (CAD), and compared PWA with the ankle-brachial index (ABI).

Methods: A trained research nurse measured PWA (radial applanation tonometry, SphygmoCor device) and ABI (hand-held Doppler) in a consecutive series of fasted patients resting supine prior to elective coronary angiography. Measurements were undertaken blind to other clinical information. Mean differences in RWT, AIX, and ABI in the presence of CAD were adjusted for age, height, mean BP, fasting cholesterol, ever smoked, and treated hypertension using multiple linear regression.

Results: We recruited 125 patients (49 women) with a mean age of 65 years, total cholesterol 4.4 mmol/l, BP 136/78, current smokers 22%, and previous myocardial infraction 30%. A statistically significant interaction between sex and CAD was present for both RWT ( p = 0.003) and AIX ( p = 0.03). No interaction was demonstrated for ABI ( p = 0.21). Mean differences for men and women in the presence/absence of CAD were: RWT −10.1 vs. +5.2 milliseconds; AIX +1.2 vs. −5.4; ABI −0.02 vs. −0.10. Male and female area under receiver operating characteristic curves for CAD detection differed for RWT (0.33 vs. 0.67) and AIX (0.62 vs. 0.36), but were similar for ABI (0.40 vs. 0.34).

Conclusion: The timing and extent of arterial wave reflections in the presence of CAD may differ in men and women.
DEOXYRIBONUCLEIC ACID CYTOPHOTOMETRY OF STAINED HUMAN LEUKOCYTES II. THE MECHANICAL SCANNER OF CYDAC, THE THEORY OF SCANNING PHOTOMETRY AND THE MAGNITUDE OF RESIDUAL ERRORS
Authors: MAYALL BRIAN H. , MENDELSOHN MORTIMER L.
Published: April 2011

The mechanical scanner of CYDAC is a dual beam cytophotometer with a Nipkow scanning disc, analogue photometric circuits, digital conversion and accumulation of the photometric values and digital control of the scanning procedure. A measuring sequence involves object and clear field scans, and the final readout is virtually independent of fluctuations and heterogeneities in illumination and of variations in the size of the scanning apertures. Optical sources of error are analyzed both theoretically and experimentally as they apply to scanning cytophotometers. The effects of conical and polychromatic illumination, stray light and heterogeneous distribution of chromophore lead to negative averaging errors that are a function of optical density. The role of scan aperture size and optical resolution on distributional error is discussed and calculated in terms of three possible models. Non-absorptive phenomena, such as scattering and diffraction, lead to positive errors that are complementary, in a qualitative sense, to distributional error. In attempting to isolate these different errors experimentally, we show that measurements of the relative deoxyribonucleic acid stain content of human leukocytes are independent, within wide limits, of condenser and objective numerical aperture and of changes in absorptivity secondary to changes in wavelength. Where an adequate theory exists, these experimental results agree well with the theoretical predictions and, in any case, they indicate that for stained leukocytes the net effect of all optical errors must be less than 2%. Random measuring errors include both photon shot noise and errors associated with the geometry of the scanning disc. These errors contribute to an error of 1.2% among replicate measurements of leukocytes made under standard conditions. The previously reported differences in stain content among leukocyte types and among individual cells within types cannot be ascribed to either optical or instrumentation errors. Thus, the measured differences must reflect actual differences either in the amount or in the absorptivity of the stain.
A HISTOCHEMICAL PROCEDURE FOR THE LOCALIZATION OF CHOLINE ACETYLTRANSFERASE ACTIVITY
Authors: BURT ALVIN M.
Published: April 2011

A histochemical procedure has been developed for the demonstration of choline acetyltransferase activity. This procedure is based upon the formation of an insoluble lead mercaptide from coenzyme A (CoASH), a product of the enzymatic reaction. Optimal conditions for the precipitation of the lead mercaptide are defined. The localization of the reaction product in the rat spinal cord is restricted to the neuronal element. Heavy deposition of reaction product in the perikaryon was noted for the anterior motor neurons; some of the other neurons were negative. Boutons terminaux were visible on the surface of all neurons studied. A combination of quantitative radiochemical data, enzyme solubilization, copper inhibition and the histologic distribution of the reaction product is discussed in relation to the specificity of the procedure for the differential localization of choline acetyltransferase and nonspecific acetyl-CoA hydrolase activities.
PROTEIN UPTAKE BY SYNOVIAL CELLS I. ULTRASTRUCTURAL STUDY OF THE FATE OF INTRAARTICULARLY INJECTED PEROXIDASES
Authors: SHANNON SARAJAYNE LIMPERT , GRAHAM RICHARD C.
Published: April 2011

The uptake of proteins in rabbit synovial membrane was studied with the peroxidase tracer technique. The morphologic characteristics of the synovial cells formed a continuum from those of type "A" to those of type "B." Cells in which type A characteristics predominated took up large amounts of protein, both by a process of bulk incorporation and by pinocytosis. Pinocytosis of smaller amounts of protein occurred in cells with characteristics of type B. Bulk incorporation in cells with type A characteristics increased with increasing molecular size of the protein tracer. Differential uptake of proteins by synovial cells may be important in determining the protein composition of synovial fluid.
ESTERASES IN DEVELOPING AND ADULT RAT LUNG II. AN ELECTROPHORETIC ANALYSIS
Authors: O'HARE KAREN HITCHCOCK , NEWMAN JOYCE K. , VATTER ALBERT E. , REISS OSCAR K.
Published: April 2011

The esterases of developing rat lung have been investigated by means of disc gel electrophoresis of lung preparations. The enzymes were analyzed by a method that uses thiol esters as substrates in order to correlate these findings with earlier biochemical and cytochemical studies. The esters, p-nitrophenylthiol acetate and p-nitrophenylthiol butyrate, were hydrolyzed by the enzymes in the presence of copper ions. The resultant copper mercaptide was stained with dithiooxamide and analyzed visually and with a densitometer. Lung preparations from prenatal (days 18 through term), postnatal (through the 4th week) and adult animals were examined for the pattern of esterases and for their sensitivity to such inhibitors as O,O-diethyl O- p-nitrophenyl phosphate (E-600) and diisopropyl phosphorofluoridate (DFP) and to temperature inactivation. The zones of esterase activity increased in number and amount during development except for two bands which decreased markedly during late prenatal development. The timing of these changes correlates with various developmental events in the rat lung.
QUANTITATIVE HISTOLOGIC DISTRIBUTION OF ADENOSINE-3',5'-MONOPHOSPHATE IN THE RAT ADRENAL AND EFFECT OF ADRENOCORTICOTROPIN
Authors: ORENBERG ELAINE K. , GLICK DAVID
Published: April 2011

Adaptations of both the enzymatic radioisotope displacement and the luminescence methods for measurement of adenosine-3',5'-monophosphate in picomole amounts in sub-milligram samples (e.g., fresh-frozen microtome sections of tissue or 10 µl homogenate) were made and applied to establishing the quantitative histologic distribution of adenosine-3',5'-monophosphate in the adrenals of untreated and adrenocorticotropic hormone-treated rats. A sharp peak in concentration of 24 pmoles/mg wet weight (1.3 pmoles/µg protein-nitrogen) was observed in the reticular zone of the untreated rats and the values fell off rapidly in the medulla to a minimum of less than one-third the peak value, and in the fasciculata and glomerulosa to less than one-half. The adrenocorticotropic hormone injections caused a second peak to appear in the outer fasciculata with a height comparable to that in the reticularis, but the concentrations in all zones were elevated. These two peaks were about 30 pmoles/mg wet weight (2 pmoles/µg protein-nitrogen), and the concentrations also fell to a minimum in the medulla, about one-third the peak values, and to relatively low values in inner fasciculata and glomerulosa, almost one-half the peak value.
THE HISTOCHEMICAL DEMONSTRATION OF NUCLEASE ACTIVITY WITH FILMS OF SOLUBLE RIBONUCLEIC ACID AND POLYADENYLIC ACID
Authors: DAOUST R. , MORAIS R.
Published: April 2011

Films of soluble ribonucleic acid (sRNA) and polyadenylic acid (poly-A) were used to investigate the distribution of nuclease activity in normal rat tissues. The reactions obtained with films of sRNA were similar to those previously observed with standard RNA and both substrates apparently reveal the same group of nucleases. On the other hand, the distribution of enzyme activity shown by films of poly-A differed markedly from that observed with RNA films, and it appears that films of poly-A demonstrate a different group of nucleases.
LOCALIZATION OF CARBONIC ANHYDRASE ACTIVITY IN TURTLE AND TOAD URINARY BLADDER MUCOSA
Authors: ROSEN SEYMOUR
Published: April 2011

Carbonic anhydrase activity was demonstrated in a distinct population of mucosal cells of whole stretched bladder of turtle and toad. The apical portions of the cells containing the enzyme were discretely stained as noncontiguous polygonal forms and allowed an estimation of the luminal surface representation of this epithelium. The slight luminal surface (0.8% or less) occupied by cells with carbonic anhydrase activity in the toad bladder contrasted with a greater representation in turtle bladder (6.4-12.2%) and correlated with the relative capacity of these tissues to acidify. After completion of the enzyme staining reaction the tissues of the turtle bladder were directly processed for electron microscopic observations without osmium tetroxide postfixation. The deposition of the electron-opaque cobalt sulfide compound was selective and was found only in cells with luminal representation and small apical granules, the recently described "third cell type"; cobalt sulfide was mainly present in the cytoplasm, an observation consonant with the known localization of carbonic anhydrase primarily in supernatant fractions.
CYTOCHEMICAL REACTIVITY OF POLLEN WALLS
Authors: SOUTHWORTH DARLENE
Published: April 2011

Cytochemical staining reactions in pollen grain walls of three species of Compositae, Ambrosia trifida, Artemisia pycnocephala and Gerbera jamesonii, were examined. The intine gives positive reactions for protein and for insoluble polysaccharides including pectic acid, callose and hemicellulose. The exine gives positive reactions for protein and for extractable lipids. In addition the structural material of the exine, sporopollenin, gives positive reactions for aliphatic double bonds and for closely packed anionic sites. Sporopollenin gives negative results to common cytochemical tests for lignin but reacts the same as lignin to metachromatic dyes. Some staining differences between exine-1 and exine-2 were noted.
PROCION YELLOW AND CATECHOLAMINE DERIVATIVES CHEMICAL RELATIONSHIPS
Authors: SIVITZ M. , KALLEN R. G. , LATIES A. M.
Published: April 2011

The use of Procion Yellow (M-4RS, ICI America, Inc.) as a fluorochrome in conjunction with catecholamine-derived isoquinoline fluorescence is examined by chemical techniques. From a series of chemical studies it has been found that: (a) Procion Yellow does not interfere with the formation of isoquinoline derivatives of catecholamine; (b) commercially available Procion Yellow is a composite of four species, all of whose emission-excitation maxima are distinguishable from that of catecholamine-derived isoquinohines; and (c) the emission-excitation maxima of Procion Yellow and of the catecholamine-derived isoquinolines are distinguishable one from another in solution. It is suggested that Procion Yellow can be a valuable fluorochrome to use in conjunction with the histofluorometric method. The data support the possibility that single adrenergic neuronal cells after being tagged with Procion Yellow can subsequently be shown to contain biogenic amines.
A NEW METHOD FOR THE INVESTIGATION OF THE KINETICS OF THE CAPTURE REACTION IN PHOSPHATASE CYTOCHEMISTRY II. THEORETICAL AND EXPERIMENTAL STUDY OF PHOSPHATE DIFFUSION FROM THIN POLYACRYLAMIDE FILMS
Authors: CORNELISSE C. J. , VAN DUIJN P.
Published: April 2011

Polyacrylamide films containing [32P] inorganic phosphate are proposed as models for the investigation of diffusion of phosphate from enzymatic sites. In connection with the use of such films for the study of metal salt capture reactions in phosphatase cytochemistry, the diffusion characteristics of the system were investigated both theoretically and experimentally. Diffusion in a theoretical model was simulated with aid of a computer program based on Fick's law. For films with a thickness above a certain value it proved possible to represent the progressive decrease in concentration by a hypothetical surface layer with zero concentration, the thickness being proportional to the square root of the product of the diffusion coefficient and time. The experimentally measured decrease in concentration showed similar characteristics. A slight deviation from the theoretical model indicated a retarding effect of an unstirred layer on the diffusion process. The apparent diffusion coefficient for inorganic phosphate with a pH of 5.0 at 25°C was estimated at 3.2 x 10–6 cm2 sec–1, but in the presence of 14% sucrose (w/v) this value was reduced to 2.3 x 10–6 cm2 sec–1. In films containing 6.7% w/v cross-linked bovine serum albumin Dapp was found to be 3.1 x 10–6 cm2 sec–1. This method offers a new approach to the accurate measurement of diffusion coefficients of small molecules in macromolecular matrices and is suitable for the study of diffusion problems in cytochemistry.
UNIFORM DISTRIBUTION AND CONCENTRATION OF AMINOPEPTIDASE IN PROXIMAL TUBULES OF PIG KIDNEY
Authors: WACHSMUTH E. D. , WOODHAMS ROSEMARY
Published: April 2011

The concentration of aminopeptidase in cross-sections in proximal tubules of pig kidney was investigated with a specific fluorescent antibody technique and a specific histochemical technique. The concentration was found to be independent of the diameter of the tubule and of the depth of the cortex. Four-micron segments of proximal tubules contained approximately l04 molecules of aminopeptidase/µ2, which is adequate for a rapid hydrolysis of peptides, even in very small concentrations, such as angiotensin II at 10 5 of the physiologic serum concentration. The findings suggest that with regard to this enzyme all sections of the proximal tubule function equivalently.
THE EFFECT OF DIGESTION WITH STREPTOMYCES HYALURONIDASE UPON CERTAIN HISTOCHEMICAL REACTIONS OF HYALURONIC ACID-CONTAINING TISSUES
Authors: YAMADA KAZUYORI
Published: April 2011

The effect of digestion with Streptomyces hyaluronidase upon certain histochemical reactions of hyaluronic acid-containing tissues has been studied in a series of human, mammalian and avian specimens. These histochemical reactions are those for the demonstration of neutral and sulfated and nonsulfated acid mucosaccharides such as periodic acid-Schiff, Alcian Blue (pH 1.0), azure A (pH 1.5), high iron diamine, aldehyde fuchsin, Alcian Blue (pH 2.5), Alcian Blue (pH 2.5)-periodic acid Schiff, azure A (pH 4.5), low iron diamine and colloidal iron. In addition, the effect of digestion with testicular hyaluronidase upon the same reactions of the same tissues was observed for comparison. Digestion with Streptomyces hyaluronidase diminishes the staining reactions due to hyaluronic acid but fails to affect those for neutral and sulfated acid mucosaccharides. These results indicate that digestion with Streptomyces hyaluronidase is a method of choice for the identification of hyaluronic acid in mucosaccharide histochemistry.
THE CYTOCHEMICAL APPLICATION OF NEW POTENT INHIBITORS OF ALKALINE PHOSPHATASES
Authors: BORGERS M.
Published: April 2011

The inhibitory properties of levamisole and its analogue R 8231 on alkaline phosphatases of various rat tissues were investigated cytochemically. Very low concentrations, respectively, 0.5 and 0.1 mM, sufficed to achieve complete inhibition in most tissues except intestine. The inhibition was reversible as for other known uncompetetive inhibitors but was found to be substrate-independent. The influence of fixation and incubation procedures was negligable. Other phosphatases as acid phosphatase, adenosine triphosphatase, thiamine pyrophosphatase, glucose 6-phosphatase and 5'-nucleotidase remained unchanged in the presence of the inhibitors.
CYTOCHEMICAL LOCALIZATION OF BRAIN NICOTINAMIDE ADENINE DINUCLEOTIDE PHOSPHATE (OXIDIZED)-DEPENDENT DEHYDROGENASES QUALITATIVE AND QUANTITATIVE DISTRIBUTIONS
Authors: SIMS K. L. , KAUFFMAN F. C. , JOHNSON E. C. , PICKEL V. M.
Published: April 2011

This study compares the histochemical and microchemical localizations of nicotinamide adenine dinucleotide phosphate (reduced) and nicotinamide adenine dinucleotide (reduced) diaphorases and four nicotinamide adenine dinucleotide phosphate (oxidized)-dependent enzymes (glucose 6-phosphate, 6-phosphogluconate, malate and isocitrate dehydrogenases) in areas of rat metencephalon and spinal cord. For the four nicotinamide adenine dinucleotide phosphate (NADP) enzymes, the pattern of localization following use of a modified tetrazolium procedure was compared with quantitative data obtained by microdissection from the same areas in adjacent sections. Optimal experimental conditions for reaction pH, temperature, substrate, cofactor and divalent cation concentrations were used for both the quantitative analysis following microdissection and the histochemical tetrazolium procedure. Consecutive sections were also examined for isocitrate dehydrogenase (nicotinamide adenine dinucleotide (oxidized)) and nicotinamide adenine dinucleotide (reduced) diaphorase activities in addition to seriatim thionine reference sections. Our results indicate that, within the central nervous system, certain characteristic qualitative differences exist in the distribution of the nicotinamide adenine dinucleotide phosphate (oxidized)- and nicotinamide adenine dinucleotide (oxidized)-dependent dehydrogenase enzymes. Nicotinamide adenine, dinucleotide enzymes are visualized predominantly in neuronal cell bodies or neuropil consisting primarily of neuronal processes; in adjacent sections, NADP enzyme activities are visualized almost exclusively in glial elements with two important exceptions. The first is the cerebellar molecular layer where the results from both micro- and histochemical techniques indicate high levels of the NADP enzymes relative to other dehydrogenases and high activity relative to the levels of these NADP enzymes in other nervous system areas. The second exception occurs in those neuronal groups known to contain high levels of catecholamines; these data are the subject of a companion report.
STAINING OF BASIC NUCLEOPROTEINS BY AN EXTRACT OF THE BLACK RASPBERRY
Authors: KASS LAWRENCE
Published: April 2011

Methanolic extracts of the black raspberry (Rubrus occidentalis) were found to stain nuclei of normal human peripheral blood and bone marrow cells. Using filter paper spot tests, staining of both purified deoxyribonucleic acid and histone occurred at pH 4.1, but at pH 9.3 only histone was stained. Histone staining diminished above pH 9.3 and disappeared at pH 10.0. Using formalin-fixed cover slips of peripheral blood and bone marrow, methanol-raspberry staining at pH 9.3 demonstrated orange-brown staining nuclei. This study suggests that the anthocyanin group of naturally occurring dyes is capable of staining basic nucleoproteins at alkaline pH.
DISTRIBUTION OF PEROXIDASE ACTIVITY IN TETRAHYMENA PYRIFORMIS MITOCHONDRIA
Authors: HIRAI KEI-ICHI
Published: April 2011

Distribution of peroxidase activity was investigated electron cytochemically by the diaminobenzidine (DAB) method in the fixed cells of Tetrahymena pyriformis strain W. Peroxidase with predominantly high activity was located in the mitochondria as shown by electron-opaque deposits of reacted DAB, but not in other cytoplasmic organelles. DAB products were deposited on mitochondrial membranes: outer membranes, inner membranes and cristate membranes. Ultrastructural sites of peroxidase activity were clearly identified on the outer sides of mitochondrial cristate membrane structures facing the intracristal spaces. Significantly, electron-dense products of DAB were also visualized on the mitochondrial inner membranes and the outer membranes. However, finer locations of peroxidase in these membrane structures were less clear. The enzyme was inactivated by heating for 10 min at 50°C or by treatment with methanol-nitroferricyanide, an inhibitor of peroxidase. Inhibition by 1 mM potassium cyanide was fairly intensive but not complete. Ten millimolar 3-amino-l,2,4-triazole, a catalase inhibitor, and 10 mM sodium azide (from neutral pH through alkaline pH) had no effects. Sodium azide, however, suppressed all peroxidase activity at pH 5. These enzymatic characteristics are similar to those of cytochrome peroxidases in the mitochondria of baker's yeast and in the cells of Thiobacillus novellus or of Pseudomonas fluorescens.
AUTORADIOGRAPHY USING DRY MOUNTED FREEZE-DRIED SECTIONS FOR LOCALIZATION OF CARCINOGENS IN THE LUNG
Authors: KENNEDY ANN R. , LITTLE JOHN B.
Published: April 2011

A freeze-dry autoradiographic technique is presented as a model for the microscopic localization of carcinogens in the lung following intratracheal injection or inhalation exposure. This totally nonliquid histologic technique minimizes carcinogen diffusion or translocation which could occur during standard tissue processing.
COMPUTER-CONTROLLED MULTIPARAMETER ANALYSIS AND SORTING OF CELLS AND PARTICLES
Authors: ARNDT-JOVIN DONNA J. , JOVIN THOMAS M.
Published: April 2011

A new high speed instrument for the multiparameter analysis and separation of cells and subcellular particles under computer control has been constructed. It incorporates several new features, the combination of which is believed to make the system uniquely capable of attacking a wide variety of biologic questions. In this paper we present the biologic objectives as well as the salient new features of the instrument.
ULTRASTRUCTURAL LOCALIZATION OF CHOLINESTERASE ACTIVITY IN THE DEVELOPING RAT RETINA
Authors: SPIRA ARTHUR W.
Published: April 2011

Retinae of rats from the 16th day of gestation to 10 weeks postnatal age were treated for the ultrastructural localization of cholinesterases according to the method of Lewis and Shute. The use of selective inhibitors served to differentiate between acetylcholinesterase and nonspecific cholinesterase activities. Nonspecific cholinesterase activity was marked in the rough endoplasmic reticulum of pigmented epithelium but only during the 1st 2 postnatal weeks. Acetylcholinesterase activity was prominent (a) in the rough endoplasmic reticulum, nuclear envelope and Golgi apparatus of ganglion cells in fetal and mature retinae; (b) transiently, between processes in the outer plexiform layer and in the perikarya of some horizontal cells; and (c) between processes in the inner plexiform layer coincident with the appearance of synapses, as well as in the mature retina. These localizations are suggestive of an association between cholinesterases and early stages of photoreceptor segment formation and consistent with a function in plexiform layer maturation and synaptic transmission in the inner plexiform layer.
Policies for Self-reliant Development
Authors: Abrol Dinesh
Published: March 2011

This article carries out an investigation into the experience of engagement with the idea of national self-reliance in the developing world during the period of last fifty years with a special focus on India's experience. It compares the experiences of implementation of the strategies of import substitution, export promotion and global integration with a view to understand their contribution to the achievement of the goals of development for a country like India during the period of last two decades. It brings out that even during the post-liberalisation period in those sectors where the policy-makers deliberately chose to delay the implementation of external liberalisation and selectively provided state protection, technological achievements have been better. However, it argues that in the post-WTO world there is the need to engineer a shift away from the exclusive reliance on either import substitution or export promotion. The strategy of development should also focus on those pathways of sustainable development that have the aim to upgrade directly the local capabilities of peasants and artisans and their access to local resources and local markets to achieve self-reliance for the people.
Histochemical localization of trehalase activity in dorsal flight muscle of the flesh fly Sarcophaga bullata with light and electron microscopy.
Authors: Chang P L , Morrison P E
Published: April 2011

Trehalase activity in flight muscle of the flesh fly Sacrophaga bullata is detected histochemically at light- and electron-microscopic levels by using diaminobenzidine, glucose oxidase and peroxidase in the incubation medium. The association of trehalase activity with the inner mitochondrial membrane is confirmed. Biochemical assay shows that about 50% of the initial total trehalase activity is lost from the tissue during the histochemical processing and about 50% remains for histochemical detection.
Simultaneous differential staining of nucleic acids, proteins, conjugated proteins and polar lipids by a cationic carbocyanine dye.
Authors: Green M R
Published: April 2011

The multiple interactions of the cationic carbocyanine dye, 1-ethyl-naptho-[1,2d]thiazolin-2-ylidene)-2-methylpropenyl]naptho[1,2d]thiazolium bromide, 'Stains-described. Many of these substances could be distinguished from one another on the basis of color in conjunction with chemical and enzymatic digestions. Further studies with this dye have shown that under certain conditions polar lipids as well may be distinguished from these substances. Stains-all has a highly sensitive metachromatic reaction for the presence of polar lipids. It is possible to detect the lipids in large part because they are green and contrast with a red- or pink-stained protein background and with the blue-purple of nuclei or cartilage. Where other green substances occur as in sialoglycoproteins of mucous or membranes, the lipids can be distinguished because they are extracted by chloroform-methanol (2:1) or pyridine.
Performance of the LARC classifier in clinical laboratories.
Authors: Cotter D A , Sage B H
Published: April 2011

As part of the installation procedure of the LARC leukocyte differential classifier in a clinical laboratory, a 100-slide protocol is carried out to establish the performance of the classifier in the laboratory. The detailed make-up of this protocol and its relationship to key performance parameters for the leukocyte differential are described in detail. Data from the first ten of these protocols are presented which establish the (a) normal ranges, (b) reproducibility, (c) accuracy, (d) false-positive/false-negative rates for the detection of left shifts and (e) false-positive/false-negative rates for the detection of bloods with abnormal cells.
Assessment of monocyte esterase activity by flow cytophotometry.
Authors: Kaplow L S , Dauber H , Lerner E
Published: April 2011

An azo dye supravital method has been devised for selectively staining human monocytes in suspension for nonspecific esterase activity. Stained cells can be identified and rapidly enumerated by presenting the suspension of stained cells to the Cytograf, a flow-through cell discriminating cytophotometer. The intensity of stain is proportional to the intracellular esterase activity. By analysis of the oscilloscope display, it has been possible to obtain relative data concerning the degree of activity of monocyte nonspecific esterase activity. These observations suggest a unique approach to the measurement of intracellular enzyme activity in selected cells in a mixed population.
Gynecologic specimen analysis by multiangle light scattering in a flow system.
Authors: Salzman G C , Crowell J M , Hansen K M , Ingram M , Mullaney P F
Published: April 2011

A flow-system instrument is described in which the laser light scattered by a mammalian cell is sampled simultaneously at up to 32 angles between 0 degrees and 21 degrees from the laser beam axis as the cell passes through the beam. The scatter pattern for each cell is stored by a computer for later analysis. Various data-processing techniques are discussed. Results of preliminary application of the instrument to the analysis of normal and abnormal gynecologic specimens are presented.
A rapid, simple and sensitive method for the demonstration of central catecholamine-containing neurons and axons by glyoxylic acid-induced fluorescence. II. A detailed description of methodology.
Authors: Bloom F E , Battensberg E L
Published: April 2011

The glyoxylic acid-induced fluorescence method for localization of brain catecholamine neurons has been modified. Fluorescence is developed rapidly in cryostat sections of brains fixed by perfusion with 0.5% depolymerized paraformaldehyde and 2.0% glyoxylic acid. Since neither freeze drying nor vibratome sectioning is required, total processing time can be less than 1 hr. Both perikarya and fine varicose axons of norepinephrine- and dopamine-containing neurons can be seen throughout the neuroaxis. The modified technique retains good cytologic integrity and may provide a useful alternative for methods combining histochemical approaches.
Biological strandard for electron microprobe analysis of intracellular sodium concentration.
Authors: Sottiurai V S , Malvin R L , Allard L F , Bigelow W C
Published: April 2011

A method is described for the determination of the intracellular concentration of sodium in individual cells using the electron microprobe analyzer. This method gives an accuracy equal to that obtained by using flame photometry on tissues with large cell populations. Intracellular sodium was precipitated in the cell by a fixative containing pyroantimonate. Cartilaginous needles from shark fins which were equilibrated in saline solutions of differing concentrations were used as biological standards.
The use of ruthenium and p-phenylenediamine to stain cartilage simultaneously for light and electron microscopy.
Authors: Shepard N , Mitchell N
Published: April 2011

p-Phenylenediamine was incorporated during the dehydration of cartilage blocks fixed with ruthenium red, glutaraldehyde and osmium. The addition of p-phenylenediamine permitted the visualization by light microscopy of cellular detail not obtained with ruthenium red alone. Ruthenium red is a widely used method for retaining and localizing proteoglycan for electron microscopy but its inability to produce sufficient density within the cells for light microscopy prevented simultaneous visualization by these two modes. The addition of p-phenylenediamine staining overcame this shorcoming without interfering with the ultrastructural detail and both stains can also be used before the making of radioautographs.
Cytochemical demonstration of 5-formyl tetrahydrofolate cyclodehydrase and 5,10-methenyl tetrahydrofolate cyclohydrolase activity.
Authors: Tzortzatou F
Published: April 2011

The enzyme 5-formyl tetrahydrofolate cyclodehydrase plays an important role in the conversion of 5-formyl tetrahydrofolate to 5,10-methenyl tetrahydrofolate. A second enzyme, cyclohydrolase, converts 5,10-methenyl tetrahydrofolate to 10-formyl tetrahydrofolate. These folate derivatives play a significant part in the biosynthesis of purines. A method has been devised for the cytochemical demonstration of 5-formyl tetrahydrofolate cyclodehydrase and 5,10-methenyl tetrahydrofolate cyclohydrolase activity which uses 5-formyl tetrahydrofolate or 5,10-methenyl tetrahydrofolate as substrate respectively, blocking possible interferences by other enzymes, and allows the nonenzymatic reduction of nitro-blue tetrazolium by 5,10-methenyl tetrahydrofolate formed by the action of the cyclodehydrase on the substrate 5-formyl tetrahydrofolate, and by 10-formyl tetrahydrofolate formed by the action of cyclohydrolase on the substrate 5,10-methenyl tetrahydrofolate, thus revealing intracellular sites of enzyme activity. The methods appear to show only intracellular localization of the blue formazan deposits of reduced tetrazolium. The distribution of positivity in cells of human blood and bone marrow is described.
Indirect fluorescence of primary and secondary myofibers in developing porcine muscle.
Authors: Beermann D H , Cassens R G
Published: April 2011

Cytochemical differentiation of two populations of developing skeletal myofibers has been demonstrated in fetal muscle with metachromatic fluorescence of ribonucleic acid and deoxyribonucleic acid by staining fresh frozed cryostat sections of developing porcine skeletal muscle with acridine orange (CL. 46005). Evidence is presented that supports the hypothesis that first-formed myofibers (primary myofibers) serve as a structural framework upon which myoblasts proliferate, fuse in linear sequence and give rise to a second population (secondary myofibers) of myofibers.
Rapid multiparameter analysis of cell stimulation in mixed lymphocyte culture reactions.
Authors: Traganos F , Gorski A J , Darzynkiewicz Z , Sharpless T , Melamed M R
Published: April 2011

A flow-cytofluorometric method, based on the differential stability of deoxyribonucleic acid versus ribonucleic acid with the metachromatic dye, acridine orange, simultaneously measures the following parameters of stimulation in mixed lymphocyte cultures: (a) number of nonstimulated cells; (b) total number of stimulated lymphocytes; (c) number of stimulated lymphocytes in G1, S and G2 + M phases of the cell cycle; (d) number of macrophages; (e) number of dead cells. The progress of lymphocyte stimulation may also be measured by a parameter representing ribonucleic acid accumulation per cell. The method is rapid, avoids cell rinsing, fixation and centrifugation and is applicable to microcultures. Multiparameter analysis of cell stimulation which provides simultaneous measurements of lymphocyte proliferation and accumulation of ribonucleic acid per cell may prove to be a more sensitive assay of histocompatibility than tests based only on cell proliferation (tritiated thymidine incorporation).
Flow cytometric analysis of bromodeoxyuridine-substituted cells stained with 33258 Hoechst.
Authors: Latt S A , George Y S , Gray J W
Published: April 2011

This paper describes a flow-cytometric application of the quenching of fluorescence from 33258 Hoechst stained Chinese hamster ovary-line cells due to the incorporation of 5-bromo-deoxyuridine (BrdU) into the cellular deoxyribonucleic acid. Cells were grown for 24 hr in medium containing BrdU in concentrations ranging from 1 x 10(-8) to 1 x 10(-4) M. For each concentration we measured the average fluorescence as determined by flow cytometry, the extent of BrdU substitution and the effect of the BrdU on cell growth. We determined that a BrdU concentration of 1 x 10(-5) M resulted in sufficient substitution to quench the fluorescence from 33258 Hoechst by a factor of 4, allowing discrimination between cycling and noncycling cells. The extent of BrdU substitution after growth for 24 hr in this concentration of BrdU was 64%. These data indicate the feasibility of detecting deoxyribonucleic acid synthesis in whole cells using the 33258 Hoechst-BrdU methodology.
Estrogen receptor immunocytochemistry.
Authors: Kurzon R M , Sternberger L A
Published: April 2011

Estrogen receptor activity was preserved in fixed, paraffin-embedded tissue and demonstrated by binding of estrogen which, in turn, was detected immunocytochemically. Estrogen was added to rat endocervial epithelium to protect specifically receptors during fixation. The protective estrogen was apparently lost during embedding and had to be resupplied before staining. Estradiol-mediated immunocytochemical staining was inhibited by diethylstilbestrol and nafoxidine hydrochloride.
A histochemical method for the identification of 9-O-acyl sialic acid. An investigation of bovine submaxillary gland and intestinal mucins.
Authors: Reid P E , Culling C F , Dunn W L
Published: April 2011

Prolongation of the initial periodate oxidation step of the periodate-borohydride technique/potassium hydroxide/periodic acid-Schiff and periodic acid-Thionin Schiff/potassium hydroxide/periodic acid-Schiff sequences produced little or no change in the diagnostic staining for the potassium hydroxide/periodic acid-Schiff effect, exhibited by the colonic epithelial mucins of man and rat and the Brunner's gland mucin of rabbits. In contrast, there was a gradual, but clear decrease in the intensity of such staining of bovine submaxillary gland mucins. It was concluded that, in the intestinal mucins studied the potassium hydroxide/periodic acid-Schiff effect was due to sialic acids bearing O-acyl substitutents at positions C7 and/or C8 whereas in bovine submaxillary gland mucin the potassium hydroxide/periodic acid-Schiff effect is probably due, at least in part, to the presence of 9-O-acyl sialic acids. This investigation has led to the development of a technique which can be used to identify 9-O-acyl sialic acids.