bluuebunny/crossref_metadata_2025 huggingface dataset:
THE EFFECT OF FAT INGESTION ON THE ESTERASE ISOZYMES OF INTESTINE, INTESTINAL LYMPH, AND SERUM
Authors: LEWIS ADAM A. M. , HUNTER ROBERT L.
Published: April 2011

The effect of various meals on the α-naphthyl butyrate isozymes of the serum and intestinal lymph has been studied in male rats using the starch gel zymogram technique. A marked increase in esterase activity, largely confined to two molecular fractions, occurred in the intestinal lymph, and later in the serum, following a meal of corn oil. The fat meal has no effect on the esterase activity of bile, although the presence of bile was essential for increased esterase activity to occur in the intestinal lymph. The source of the isozymes most affected by ingested fat is considered to be the small intestine, particularly the duodenum.
A SIMPLE METHOD FOR THE DETERMINATION OF WATER CONTENT AND PROTEIN CONCENTRATION IN SUBCELLULAR ELEMENTS
Authors: GLAS U. , BAHR G. F.
Published: April 2011

An accurate method for the determination of the average water content of subcellular particles from their suspensions is presented. The particle is freed from water by spinning it from its suspension medium into oil of high density. In the oil it is broken up, and the water is distilled off for its gravimetric determination in an absorption tube. The dry particle remnants are further centrifuged back into a watery medium in which the nitrogen content is determined A. protein content of 26.3% for the average rat liver mitochondrion is found.
NORMAL HUMAN NEUTROPHIL LEUKOCYTES AS A REFERENCE SYSTEM FOR THE MICROSPECTROPHOTOMETRICALLY QUANTITATED PERIODIC ACID-SCHIFF REACTION
Authors: GAHRTON GÖSTA
Published: April 2011

Normal human neutrophil leukocytes are suggested as references for a microspectrophotometrically quantitated PAS reaction in biological material. Analysis of variance of values for the total extinction at 546 mµ in PAS-stained, single, normal neutrophils proved these cells to be suitable references. Methanol-fixed, smeared neutrophils could be stored for several months without changing their amount of PAS reactive material. A person with a mean for the amount of PAS reactive material in neutrophils that lay close to the average in a normal material of 20 persons was chosen as the original reference subject. New reference series of neutrophils were related to the original one, thus obtaining a continuous reference system. The results are based upon microspectrophotometric measurements of about 3500 individual neutrophils.
AN APPROACH TO THE QUANTITATIVE HISTOCHEMISTRY OF THE A-CELLS IN THE PANCREATIC ISLETS
Authors: BROLIN SVEN E. , OHLSSON ARNE , BORGLUND ERIK
Published: April 2011

Our present knowledge of the quantitative histochemistry of the pancreas is mainly related to the enzymatic properties of the exocrine parenchyma and the B-cells. Appropriate analyses of A-cells require their isolations, which can be accomplished in animals with different islet cells located in separate groups. In fresh frozen sections of the pancreas from ducks groups of A- and B-cells were recognized by darkfield microscopy combined with ordinary microscopic examination after fixation and staining. Each adjacent section was lyophilized and separate samples of acini, A-and B-cells were dissected by free hand. Using quantitative microchemical techniques malic dehydrogenase, lactic dehydrogenase and glutamic-oxaloacetic transaminase activities were measured. The enzymatic distribution found between samples of B-cells and acini was in accordance with earlier findings in mammals, as B-cells showed higher activities except for lactic dehydrogenase. The A-cells differed from the B-cells by lower activities of malic dehydrogenase and glutamic-oxaloacetic transaminase and from the acini by lower activities of lactic dehydrogenase.
QUANTITATION OF TISSUE-BOUND RENAL AMINOPEPTIDASE BY A MICRODENSITOMETRIC TECHNIQUE
Authors: FELGENHAUER K. , GLENNER G. G.
Published: April 2011

Relationships between biochemical and histochemical assay systems can be evaluated by appropriate techniques. With rat kidney as the test object the enzyme localized in the tissue section hydrolyzing l-leucyl-β-naphthylamide (LNA) was found to be identical with a purified particle-bound, cobalt-activated aminopolypeptidase previously characterized biochemically. A soluble sulfhydryl-dependent aminopeptidase, present in the rat kidney and also known to hydrolyze LNA, was not demonstrated in the histochemical system, although the diazonium salt employed incidentally caused the insolubilization in the tissue section of a significant proportion of previously extractable, particle-bound enzyme. Linearity between the enzymic activity present and the measurable enzymic activity was demonstrated to exist at the substrate concentration, pH level, section thickness and diazonium salt concentrations used in the assay procedure, but did not exist when these factors were varied beyond certain well defined limits. In addition, the effect of diazonium salt on both the inhibition and insolubilization of enzyme in the tissue section could be quantitated. Based on this evaluation of the enzymes hydrolyzing LNA in the rat kidney, a microdensitometric assay method employing a constant flow incubating chamber was developed to characterize and quantitate the LNA-hydrolyzing enzyme in the proximal tubuli. This study defines many of the parameters necessary for the future investigation by quantitative and qualitative methods of histochemical systems capable of providing resolutions of a higher order of magnitude and the retention of a greater proportion of extractable enzyme within the tissue section.
DISTRIBUTION PATTERNS OF OXIDATIVE ENZYMES IN EXPERIMENTALLY INDUCED TUMORS, REGENERATION AND INFLAMMATION OF MOUSE SKIN
Authors: HASHIMOTO TAKENORI , BURSTONE M. S.
Published: April 2011

Enzyme histochemical studies of normal and burned mouse skin as well as those treated with methylcholanthrene or 2,4-dinitro-1-chlorobenzene revealed that the irritant (or stimulus) carcinogenic or noncarcinogenic, initially results in a more distinct polarization2 of the localization of oxidative enzymes including glucose 6-phosphate dehydrogenase (predominant in the upper layers of the hypertrophic epidermis) and DPN-dependent dehydrogenases, succinic dehydrogenase and cytochrome oxidase in the lower layers. Subsequently, only the carcinogen disrupted the polarization pattern. Methylcholanthrene-induced squamous cell carcinomas were classified from a histochemical standpoint according to the following three types: (1) those well differentiated and having a distinctly polarized pattern of oxidative enzymes; (2) those showing disruption of the polarization pattern and marked activity of oxidative enzymes; and (3) those that were undifferentiated and characterized by absence of a polarized pattern and weak activity of oxidative enzymes. Type 1 showed an enzyme distribution pattern resembling that of the inflammatory hypertrophic epidermis; type 2 was similar in enzymatic pattern to the advancing proximal portion of the regenerating epidermis; and type 3 was similar enzyme-histochemically to the undifferentiated epithelium of the early embryo skin. Thus, a great variation in the distribution pattern of enzyme activities appears during the carcinogenesis process.
LEAD ION AND PHOSPHATASE HISTOCHEMISTRY I. NONENZYMATIC HYDROLYSIS OF NUCLEOSIDE PHOSPHATES BY LEAD ION
Authors: ROSENTHAL ALAN S. , MOSES HAROLD L. , BEAVER DAVID L. , SCHUFFMAN SHIRLEY S.
Published: April 2011

A previously undescribed catalytic action of lead ion on the nonenzymatic hydrolysis of nucleoside phosphates has been demonstrated. Lead ion (3.6 mM) hydrolyzed adenosine triphosphate (ATP) at pH 7.2 and 37°C. The presence of magnesium and imidazole was stimulatory. The rate appeared to increase with temperature from 25-60°C. The reaction was inhibited by ethylenediaminetetraacetate. Other nucleoside phosphates were hydrolyzed, some less rapidly than ATP. Adenosine triphosphate in the presence of a rate-limiting amount of lead acted as an inhibitor at high concentrations. With a rate-limiting concentration of ATP (0.72 mM), increasing concentrations of lead ion above 0.36 mM catalyzed a linear increase in ATP hydrolysis. It is suggested that this reaction may be a source of artifact in the lead salt method for the histochemical localization of nucleoside phosphatases.
A CONGO RED STAINING METHOD FOR EPOXY-EMBEDDED AMYLOID
Authors: SHIRAHAMA TSURANOBU , COHEN ALAN S.
Published: April 2011

A Congo red staining method for osmium-fixed, epoxy-embedded tissues is described. The staining is carried out by floating thick sections (0.5-2.0 µ) upon a drop of 0.5-1.0% Congo red in 0.1 N sodium hydroxide at 45°C for 60-120 min. The amyloid, which stains a clear red, shows characteristic green birefringence when viewed in the polarization microscope. This staining method is useful for identification and mapping of amyloid substance in serial thick sections and for sampling in preparation for the electron microscopic studies of adjacent thin sections.
A MORPHOLOGIC AND CYTOCHEMICAL STUDY ON THE GREAT ALVEOLAR CELL
Authors: SOROKIN SERGEI P.
Published: April 2011

Lungs from marsupials, bats and rodents were studied by light and electron microscopy. In all three groups, the great alveolar cells exhibit similar morphologic and cytochemical characteristics. Cytoplasmic vacuoles seen in these cells by light microscopy correspond to cytosomes that are demonstrable in them by electron microscopy. Such cytosomes are osmiophilic, periodic acid-Schiff-positive and stainable with Sudan black after acetone extraction. After fixation in a mixture of aldehydes, followed by extraction in chloroform-methanol and postfixation in osmium tetroxide, cytosomes lose their osmiophilia. The cytoplasm of the great alveolar cell is notable for a loosely ordered granular endoplasmic reticulum, an extensive Golgi apparatus and numerous multivesicular bodies. Many forms transitional in appearance between multivesicular bodies and cytosomes are present. In these, osmiophilic matter occupies the intervesicular space. It is proposed that these bodies are the precursors of cytosomes. The cytosomes are interpreted to be products of the "lysosomal" system in this cell. Ultimately they are secreted onto the alveolar surface.
EVALUATION OF THE GLYCOGENOLYTIC EFFECT OF α-AMYLASE USING RADIOAUTOGRAPHY AND ELECTRON MICROSCOPY
Authors: COIMBRA A.
Published: April 2011

The effectiveness of crystalline α-amylase and saliva in hydrolyzing newly formed glycogen in liver and muscle was examined. Glycogen synthesis was induced by the administration of H3-glucose to fasting rats or by the incubation of tissue slices in a medium containing H3-glucose. Paraffin sections of Rossman-fixed tissues or small pieces of liver fixed in glutaraldehyde and subsequently postosmicated and embedded in Epon were then enzymatically digested. Grain counts were made in radioautographs of treated and untreated materials, and the amount of radioactivity removed by the digestion was used to assess the efficiency of the enzymes in hydrolyzing glycogen. Crystalline α-amylase hydrolyzed almost completely newly formed glycogen in liver and muscle. Saliva removed the glycogen that was synthesized in vivo, but it was less effective in hydrolyzing glycogen synthesized in vitro. Electron micrographs of digested liver cells confirmed the radioautographic findings on the effectiveness of the enzyme preparations.
THE NUMBER OF ACETYLCHOLINESTERASE MOLECULES IN THE RAT MEGAKARYOCYTE
Authors: DARŻYNKIEWICZ ZBIGNIEW , ROGERS ANDREW W. , BARNARD ERIC A.
Published: April 2011

A megakaryocyte cell series from rat bone marrow has been examined by the isotopic di-isopropyl fluorophosphate (DFP) method for esterases. After complete reaction with 32P-DFP, the numbers of DFP-reacted molecules in individual cells have been determined by β track auto-radiography. Previous work has shown the percentage of organophosphate-sensitive sites in these cells which can be taken as active centers of acetylcholinesterase (AChase). Combining these data, the absolute numbers of organophosphate-sensitive esterase molecules and AChase molecules per cell were determined. Histograms show a narrow spread of values within each of four size classes from megakaryoblast to fully mature megakaryocyte, but, with means increasing 4-fold through this series, approximately in proportion to cell volume. A rat megakaryoblast has 2 x 106 AChase molecules, and a megakaryocyte (of 48-µ diameter) has 7.6 x 106 molecules. The apparent turnover number of the enzyme for intracellular reaction with substrate is calculated and compared with turnover numbers available for other AChases.
A SEMIAUTOMATIC INSTRUMENT FOR THE RADIOAUTOGRAPHIC COATING TECHNIQUE
Authors: KOPRIWA BEATRIX MARKUS
Published: April 2011

By means of a mechanical coating instrument a fast, simple method to coat specimens with liquid nuclear track emulsion has been devised for quantitative light and electron microscopic radioautography. In both cases, the section is mounted on a glass slide. After the vertically held slide has been immersed in the melted emulsion, the instrument withdraws it at a slow, constant speed. As a result, the specimen is coated with a thin, uniform emulsion layer composed of homogeneously distributed silver bromide crystals. The thickness of the emulsion coat may be standardized by selection of an optimal combination of emulsion dilution, temperature and withdrawal speed.
A REVIEW OF EARLY CONCEPTS OF AMYLOID IN CONTEXT WITH CONTEMPORARY CHEMICAL LITERATURE FROM 1839 TO 1859
Authors: PUCHTLER HOLDE , SWEAT FAYE
Published: April 2011

Early concepts of amyloid are reviewed in context with chemical literature from 1839-1859. Cellulose was first described in 1839. Treatment with sulfuric acid converted cellulose into a substance which was colored blue by iodine. This cellulose derivative was called amyloid. Virchow applied the chemical tests for starch and cellulose to human tissues. The substance in waxy degeneration reacted like immature cellulose. Virchow did not coin the term amyloid for this substance but suggested it as a compromise. Virchow restricted the term amyloid to the carbohydrate moiety and explicitly excluded the protein fraction which he had described earlier.

Friedreich and Kekulé (1859) studied an evidently impure sample of amyloid and concluded that it did not contain polysaccharides. Since then the protein moiety has been studied extensively, but the carbohydrate fraction has been largely neglected. It appears possible that investigations of the carbohydrates at sites of amyloid formation would aid the understanding of the pathogenesis of amyloidosis2.
A NEW ION ASSOCIATION-FRACTIONATION TECHNIQUE FOR IDENTIFYING INDIVIDUAL MUCOPOLYSACCHARIDES AND OTHER POLYANIONS IN TISSUE SECTIONS
Authors: ZUGIBE FREDERICK T. , FINK MARILYN L.
Published: April 2011

A new histochemical ions association-fractionation technique has been developed for identifying individual mucopolysaccharides and other biological polyanions in tissue sections. This method is based on the differential precipitation of individual polyanions with quaternary ammonium salts in solutions of specific ionic concentration and the subsequent formation of red ion association complexes with the bound quaternary moieties. Hyaluronic acid, the chondroitin sulfates, heparitin sulfate, heparin and RNA were readily identified. Further differentiation was effected by incorporating acetone-dried powder extracts from Flarobacterium heparinum which has been adapted to chondroitin sulfate and heparin.
HUMAN PITUITARY LUTEINIZING HORMONE: AN IMMUNOHISTOCHEMICAL STUDY
Authors: MIDGLEY A. REES
Published: April 2011

Luteinizing hormone (LH) was localized immunohistochemically to cells in the pars anterior and pars tuberalis of the human pituitary gland. Upon restaining with a variety of histochemical procedures, LH-containing cells were found to be aldehyde fuchsin and periodic acid-Schiff (PAS) positive; however, the intensity of histochemical staining failed to correlate with that of fluorescence. Only a few of the PAS- or aldehyde fuchsin-positive cells contained LH. In contrast to the above, with the performic acid-alcian blue-PAS-organge G sequence of Adams, all cells containing LH were turquoise colored, alcian blue-positive (S1 mucoid cells) and vice versa. This study was performed with antiserum specific for human chorionic gonadotropin which was shown to cross react in immunodiffusion analysis with purified human pituitary luteinizing hormone and to be capable of neutralizing its biological activity.
LYSOZYME IN BONE MARROW AND PERIPHERAL BLOOD CELLS
Authors: BRIGGS ROBERT S. , PERILLIE PASQUALE E. , FINCH STUART C.
Published: April 2011

By means of an indirect histochemical technique, the intracellular lysozyme of the formed elements of the peripheral blood and bone marrow was estimated. Evidence is presented that monocytes, as well as mature neutrophils and their precursors extending back to the progranulocyte, contain significant amounts of this enzyme. A rare mature eosinophil demonstrated a trace of lysozyme activity. There was no evidence of lysozyme activity in basophils, erythrocytes, megakaryocytes, platelets, plasma cells, tissue mast cells or bone marrow reticuloendothelial cells.
A NEW HISTOCHEMICAL METHOD FOR ACID PHOSPHATASE BY THE USE OF 5-IODOINDOXYL PHOSPHATE
Authors: EVANS G. W. , WHINNEY CECILIA L. , TSOU K. C.
Published: April 2011

5-Iodoindoxyl phosphate has been found to be a useful indigogenic substrate in the histochemical demonstration of acid phosphatase activity. Its superiority to other indoxyl phosphates is apparently due to a rapid oxidation of 5-iodoindoxyl to 5,5'-diiodoindigo in the acid pH range. A redox system of ferri-ferrocyanide enhances the oxidation and improves the localization. This method can be applied to calcium-formol-fixed tissues or to fresh frozen sections, although fixed tissues yield better results. The method is not recommended for the demonstration of enzyme activity in lipid-rich tissues because of the complexing property of lipids with 5,5'-diiodoindigo that results in crystal formation. The distribution of acid phosphatase activity with this method is generally similar to that obtained using azo dye methods.
HYPERTROPHY OF THE AGRANULAR ENDOPLASMIC RETICULUM IN HAMSTER LIVER INDUCED BY PHENOBARBITAL (WITH A REVIEW ON THE FUNCTIONS OF THIS ORGANELLE IN LIVER)
Authors: JONES A. L. , FAWCETT D. W.
Published: April 2011

The electron microscopic appearance of normal hamster liver is illustrated, and attention is drawn to certain unusual features of hepatocyte fine structure that have not hitherto been described. Examination of liver cells from animals receiving repeated intraperitoneal injections of phenobarbital, reveals a spectacular hypertrophy of the agranular reticulum and a depletion of particulate glycogen. The increase in smooth endoplasmic reticulum cannot be explained by loss of ribosomes from the granular reticulum and is regarded as an absolute gain of new membrane. Numerous examples of continuity between granular and agranular reticulum are observed, and it is suggested that protein synthesis associated with the granular reticulum contributes to the formation of the new smooth-surfaced membranes. In discussing the morphological alterations of the liver induced by phenobarbital, numerous biochemical changes reported in the literature have been reviewed in an effort to gain a clearer view of the over-all significance of the agranular reticulum in liver cell function. The evidence points to an important role in hydroxylation and elimination of lipid-soluble drugs, and in cholesterol and steroid hormone metabolism. There are indications that it also plays some role in carbohydrate metabolism, but the exact nature of its participation cannot yet be defined.
LOCALIZATION OF ACID PHOSPHATASE ACTIVITY AND SECRETION MECHANISM IN RABBIT PANCREATIC B-CELLS
Authors: LAZARUS SYDNEY S. , VOLK BRUNO W. , BARDEN HERBERT
Published: April 2011

Utilizing formaldehyde- or glutaraldehyde-fixed tissue and Gomori's lead method it was found by optical microscopy that rabbit pancreatic islet cell acid phosphatase activity is present in discrete, mostly perinuclear foci and that this distribution differs from that of the aldehyde fuchsin-positive secretory granules which are densely packed at the capillary pole of the cell. Electron microscopically lead reaction product was noted in dense bodies, as well as in structures thought to be Golgi vacuoles and vesicles, it was also present in the innermost of the Golgi cisternae, and at the periphery of adjacent single membrane-limited bodies whose origin can be traced from the proximal cisternae. These latter bodies in routinely prepared, osmium-fixed material show finely granular content, which is in contrast to the electron-dense, central body seen in secretory granules that appear to originate from endoplasmic reticulum. B-cell cytoplasm contained additional numerous, single membrane-limited vacuoles with pale content. These are thought also to represent secretion vacuoles but with insulin secretory product in a different physical or chemical state. The lack of acid phosphatase activity in B-cell secretion vacuoles, the dissimilarities in fine structure between the content of secretory elements and that of the Golgi-derived granular body, together with previous evidence that alteration in B-cell functional state does not result in altered number or distribution of acid phosphatase active elements in B-cell cytoplasm, indicate a lack of relationship between acid phosphatase and secretory granule formation or release in pancreatic B-cells. It is also hypothesized that the secretory vacuole with central dense granule may be a storage form while the pale vacuole is the one which liberates its content to the intercellular space.
ENZYMORPHOLOGIC DEMONSTRATION OF GLUCOSE 6-PHOSPHATE-DEPENDENT GLYCOGEN SYNTHETASE IN MOUSE LIVER
Authors: SIE HSIEN-GIEH , SAWYER DEBORAH , FISHMAN WILLIAM H.
Published: April 2011

Mouse liver possesses G-6-P-dependent glycogen synthetase and not the G-6-P-independent variety. This fact explains the higher requirement (450mg per 25 ml) for G-6-P in the incubation mixture for staining mouse liver glycogen synthetase than that present in the Takeuchi and Glenner substrate mixture. The higher G-6-P content, as well as other considerations, necessitated the establishment of new conditions for the staining reaction for glycogen synthetase, i.e., pH 9.4, UDPG concentration 200 mg per 25 ml, etc. The presensce of NaF (4.8 x 10–2 M) prevents thermal inactivation of glycogen synthetase at 37°, which is the temperature now recommended for the staining reaction. Postfixation and dehydration makes the sections more readily interpretable and prolongs markedly the life of the iodine staining reaction. The new procedure is reproducible and consistent in serial sections of the same animal and among different animals of like glycogen content. The enzyme appears to be located on membranes as well as in granules. Some similarities between the characteristics of the dependent and independent forms of the enzyme, as we and others have observed them by both histochemical and biochemical techniques, are discussed.
MODIFIED PROCEDURE FOR THE HISTOCHEMICAL LOCALIZATION OF RIBONUCLEASE ACTIVITY BY THE SUBSTRATE FILM METHOD
Authors: DAOUST ROGER
Published: April 2011

The substrate film method for localizing RNAse activity has been the object of further studies, and various steps of the original procedure have been modified. The modifications concern mainly the type of RNA used as substrate and the preparation of gelatin-RNA films. Moreover, a technique proposed by other authors for exposing films to tissue sections was found to present interesting advantages and has been adopted. The modified procedure proved more convenient than the original method, and a higher resolution was achieved in the present work. This report describes the revised procedure for localizing RNAse activity in tissue sections, exposes the advantages of the various modifications and presents the results obtained with different rat tissues, namely, the testis, intestine, brain, kidney, pancreas and ovary.
THF EARLY STAGES OF ABSORPTION OF INJECTED HORSERADISH PEROXIDASE IN THE PROXIMAL TUBULES OF MOUSE KIDNEY: ULTRASTRUCTURAL CYTOCHEMISTRY BY A NEW TECHNIQUE
Authors: GRAHAM RICHARD C. , KARNOVSKY MORRIS J.
Published: April 2011

The early stages of absorption of intravenously injected horseradish peroxidase in proximal tubules of mouse kidney were studied with a new ultrastructural cytochemical technique. In animals killed as early as 90 sec after injection, reaction product was found on the brushborder membranes and in the apical tubular invaginations. From the latter structures it was transported to the apical vacuoles, in which it was progressively concentrated to form protein absorption droplets. The method, which employs 3,3'-diaminobenzidine as oxidizable substrate, gives sharp localization and is sensitive. This system is advantageous in studying the early stages of renal tubular protein absorption, since small amounts of protein on membranes and in tubules and vesicles can be detected easily. The method also appears promising for studying protein transport in a variety of other cells and tissues.
PHOSPHOTUNGSTIC ACID-HEMATOXYLIN. REACTIVITY IN VITRO
Authors: TERNER JACOB Y.
Published: April 2011

In vitro, phosphotungstic acid-hematoxylin undergoes a bathochromic change in its absorption spectrum comparable to the characteristic bathochromic metachromasia observed in stained tissues when it is reacted with appropriate excesses of organic basic groups such as amines, guanido and imidazole groups. The qualitative characteristic of the reaction is a shift from the PTA-H lake's usual absorption maximum of 560 mµ to 620 mµ. Quantitatively, a maximal reaction is obtained at an organic base: PTA-H ratio of 8:1; the quality of the base does not effect the reaction so that the eight bases could be all amines, four amines and four imidazoles, etc. Akylated amines and histone extracts form blue, bathochromic precipitates with PTA-H. These data delineate the anionic character of this lake and the nature of its metachromatic properties.
MICROSPECTROPHOTOMETRIC ANALYSIS OF BASIC PROTEIN RICH SITES STAINED WITH BIEBRICH SCARLET
Authors: DOUGLAS STEVEN D. , SPICER SAMUEL S. , BARTELS PETER H.
Published: April 2011

Microspectrophotometric analyses on histologic sections stained with the acid dye Biebrich scarlet at pH 9.6 and above reveal absorption curves with maxima at 505 mµ for pink-red orthochromatic sites. Such sites of strongly basic protein include particularly spermatozoa, eosinophil and heterophil granules, Paneth cell granules, elastic fibers and seminal vesicle fluid. In contrast, absorption curves of sites with a metachromatic red-orange staining have a maximium of 485 mµ. The structures containing a strongly basic metachromatic protein invariably are the sites of a polyanion with metachromatic staining properties. Such sites include nuclei, mast cell granules, ovarian follicular fluid and various mucous acini and goblets. The absorption spectra of orthochromatic and metachromatic sites in tissue sections correspond with the spectra of Biebrich scarlet in solution alone and with polyanions, respectively. Similarly, histologic structuures containing acid mucosaccharides when stained with the basic dye safranin O at low pH displayed absorption maxima at 530 mµ pink-red orthochromatic sites and at 500 mµ for red-orange metachromatic sites.
QUANTITATIVE ANALYSIS OF THE ESTERASES IN RAT MEGAKARYOCYTES BY A RADIO-ISOTOPE CYTOCHEMICAL TECHNIQUE
Authors: DARŻYNKIEWICZ Z. , ROGERS A. W. , BARNARD E. A.
Published: April 2011

Megakaryocytes from bone marrow of immature rats have been treated with 3H-DFP, and the grain density over them measured in autoradiographs. The uptake of labeled DFP is reduced by one-third by eserine and by the AChase inhibitor 284C51, and by reactivation with pyridine-2-aldoxime. It is concluded that one-third of the DFP taken up by megakaryocytes represents binding to true AChase. The other esterases present include no significant amount of pseudocholinesterase.
HISTOCHEMICAL DETECTION OF TRIGLYCERIDE ESTERS WITH SPECIFIC LIPASES AND A CALCIUM-LEAD SULPHIDE TECHNIQUE
Authors: ADAMS C. W. M. , ABDULLA Y. H. , BAYLISS O. B. , WELLER R. O.
Published: April 2011

A histochemical method for triglyceride esters is described that depends on hydrolysis of triglycerides to fatty acids by pancreatic lipase, precipitation of the released fatty acid as calcium soap and formation of lead sulphide by the conventional Gomori technique. The specificity of the lipase for triglyceride was investigated by chromatography and activation-inhibition studies. The enzyme preparation used hydrolyzed triglycerides and waxes but did not split cholesterol esters or phosphoglycerides. Positive histochemical reactions were obtained with this pancreatic lipase method in medium sized and fine lipid droplets in frozen sections. Positive reactions were observed in rat brown fat; in various adipose tissues in man, rabbit and rat; in the spermatogonia of rat testis; in human epidermis and sebaceous glands; in solitary unidentified adrenocortical cells in man and rat; in human fatty liver and in fine droplets in senile human myocardium. A slight variable reaction was obtained in human atherosclerotic plaques. The reaction of brown fat was also investigated with the electron microscope. No triglyceride was demonstrated in unfixed tissue sectons when an active preparation of lipoprotein lipase was used in place of pancreatic lipase. This failure was probably due to diffusion of lipoprotein from unfixed sections. Lipoprotein lipase is inactive against serum lipoproteins fixed with formaldehyde or glutaraldehyde.
THE ENZYMATIC HYDROLYSIS OF AMINO ACID β-NAPHTHYLAMIDES II. PARTIAL PURIFICATION AND PROPERTIES OF A PARTICLE-BOUND COBALT-ACTIVATED RAT KIDNEY AMINOPEPTIDASE
Authors: FELGENHAUER K. , GLENNER G. G.
Published: April 2011

In order to determine the relationship and characteristics of the soluble and tissue-bound (so-called "lyo" and "desmo") components of enzymes in histochemical and biochemical systems, a biochemical investigation of the hydrolysis of l-leucyl β-naphthylamide in rat kidney was undertaken. By ultracentrifugation, autolysis, salt and solvent fractionation procedures and DEAE-cellulose column chromatography primary soluble and tissuue-bound enzymes were separated and partially purified. The soluble enzyme was found to be inhibited by p-chloromercuribenzoate and reactivated by cysteine. The tissue-bound enzyme was found to be inhibited by o-phenanthroline and reactivated by Co++, and hydrolyzed peptides and amino acid β-naphthylamides at rates differing markedly from those of the soluble enzyme and commercial leucine aminopeptidase. Disc electrophoresis of the tissue-bound enzyme preparation demonstrated two protein bands, both revealing enzymic activity with almost identical hydrolytic characteristics, i.e., isozymes. This evidence established the identity of the tissue-bound Co++-activated enzyme with that found histochemically in the brush border of the proximal tubuli, and indicated its functional capacity as that of an aminopolypeptidase. It was also concluded that in the system investigated the "lyo" and "desmo" components represented, in reality, distinct enzymes with unique characteristics.
WATER-SOLUBLE PLASTIC MOUNTING MEDIA
Authors: MCCURDY L. E. , BURSTONE M. S.
Published: April 2011

Several new water-soluble mounting media derived from polyvinyl alcohol and acrylic resins have been reported. These are of use in the preservation of various azo dye products, fats and histological stains. They may also offer an approach to new embedding techniques.
COMPARISON OF THE GOLD-THIOCHOLINE AND GOLD-THIOLACETIC ACID METHODS FOR THE HISTOCHEMICAL LOCALIZATION OF ACETYLCHOLINESTERASE AND CHOLINESTERASES
Authors: KOELLE GEORGE B. , GROMADZKI CORNELIA G.
Published: April 2011

The thiocholine (ThCh) and thiolacetic acid (ThAc) methods for the histochemical localization of acetylcholinesterase (AChE) and cholinesterases (ChE's) were modified to permit the substitution of aurous gold for copper and lead, respectively, as the capturing agent. The chief advantage of the modifications is that the precipitates formed (AuThCh-phosphate and AuS) have fine, colloidal dimensions and high electron density, thus improving their potential usefulness as electron microscopic methods. By means of selective inhibitors of AChE (10–5 M BW 284), of ChE's ( 10–7 M DFP or 3.10–8 M Nu-683), and of both enzymes (10–5 M DFP or eserine), it was shown that the gold thiocholine (AuThCh) method retains the high specificity of the original ThCh method, whereas the gold thiolacetic acid (AuThAc) method, like the lead ThAc procedure, is considerably less specific. However, the AuThAc method appears to permit finer localization than the AuTuCh technique.
HISTOCHEMICAL IDENTIFICATION OF NORADRENALINE IN FLUORESCENCE MICROSCOPY BY BOROHYDRIDE-PERIODIC ACID SEQUENCE
Authors: MUKHERJI MRITYUNJOY , RAY ANJAN KUMAR , SEN PARIMAL BIKAS
Published: April 2011

The catecholamines—dopa, dopamine, adrenaline and noradrenaline—are known to produce fluorescing 3,4-dihydroisoquinoline derivatives on condensation with formaldehyde. These dihydro products could be converted to 1,2,3,4-tetrahydroisoquinoline derivatives by borohydride reduction and in turn again to dihydro forms by re-exposing to formaldehyde vapor. Periodic acid was found to cleave the —C—C—bond at 3—4 position of tetrahydroisoquinoline derivative of noradrenaline only. This prevented the cyclitization of heterocyclic ring of tetrahydro form to give fluorescing dihydro product. This reaction can be utilized for the localization of noradrenaline in histochemical fluorescent preparations. Noradrenaline is prevented from regaining its fluorescence on re-exposure to formaldehyde by imposing periodic acid oxidation after borohydride bleaching.
HISTOCHEMICAL LOCALIZATION OF CATECHOLAMINES IN THE URINARY BLADDER OF THE TOAD (BUFO MARINUS)
Authors: MCLEAN J. R. , BURNSTOCK G.
Published: April 2011

Fluorescent histochemical localization of monoamines in whole mounts of the bladder of the toad ( Bufo marinus) revealed bundles of fluorescent nerves containing predominantly adrenaline. These formed a wide meshed net work about arteries but not veins. A small proportion of muscle bundles was supplied by fine varicose fluorescent nerves. Loading with adrenaline or noradrenaline increased the intensity of the fluorescence in the nerves, but produced no appreciable increase in the number of muscle bundles supplied by fluorescent nerves. Large, brilliantly fluorescent cells containing a primary monoamine were located along nerve bundles. Chronic reserpine treatment abolished the fluorescence in nerves but not in these cells. Ganglion cells showed no specific fluorescence, nor did fluorescent terminals impinge on them, but many ganglion cells contained autofluorescent granules. The significance of the presence and localization of catecholamines in the toad bladder is discussed in relation to the pharmacology of the organ and to histochemical investigations of the innervation in other vertebrate bladders.
HISTOCHEMICAL AND BIOCHEMICAL ANALYSIS OF THE NONSPECIFIC ESTERASES OF THE SMALL INTESTINE OF THE RAT
Authors: FRIEDMAN BRUCE , STRACHAN DONALD S. , DEWEY MAYNARD M.
Published: April 2011

With α-naphthyl butyrate and α-naphthyl acetate as substrates nonspecific esterase activity was demonstrated in both the crypt epithelium and epithelium covering the villus of the small intestine of the rat. This distribution of enzymatic activity does not correspond to the distribution of the so-called "absorptive epithelium." Starch gel electrophoretic analysis of intestinal homogenates showed a spectrum of enzymatically active fractions which hydrolyzed both substrates. The number of enzymic fractions and their electrophoretic mobilities were the same with both substrates, but the butyrate ester was hydrolyzed 2.5 times more rapidly than the acetate ester. These observations are discussed in relation to the rapid differentiation which occurs as cells migrate from the intestinal crypts onto the surface of the villus.
AN IMPROVED METHOD OF FIXATION FOR FORMALIN-SENSITIVE ENZYMES WITH SPECIAL REFERENCE TO MYOSIN ADENOSINE TRIPHOSPHATASE
Authors: HAYASHI MASANDO , FREIMAN DAVID G.
Published: April 2011

Studies have been made on the effect of fixatives containing substrate and other additives on the histochemical staining reaction for myosin adenosine triphosphatase activity in the leg muscle, and α-glycerophosphate and succinate dehydrogenase activities in the kidney of the rat. Myosin adenosine triphosphatase was well preserved in the presence of adenosine triphosphate in fixative prepared from methanol-free formaldehyde and buffered to pH 7.2 with cacodylate. Addition of sucrose was found to increase the enzyme preservation. Similar protection of the enzyme activity was afforded by other polyphosphates, particularly sodium pyrophosphate and, to less degree, thiamine pyrophosphate, adenosine diphosphate and uridine triphosphate. α-Glycerophosphate dehydrogenase was also preserved to greater degree by a similar fixative containing α-glycerophosphate than by substrate-free fixative. Succinate dehydrogenase was not significantly preserved in succinate-containing fixative. On the other hand, sucrose in the fixative increased the preservation of succinate dehydrogenase but had no appreciable effect on α-glycerophosphate dehydrogenase.
COMPARISON OF RED AND WHITE VOLUNTARY SKELETAL MUSCLES OF SEVERAL SPECIES OF PRIMATES
Authors: BEATTY C. H. , BASINGER G. M. , DULLY C. C. , BOCEK R. M.
Published: April 2011

There was a direct correlation between the qualitative histochemical classification by staining intensity for succinic dehydrogenase and the quantitative measurements of succinic dehydrogenase activity for the quadratus femoris (red), soleus (red), sartorius (predominantly red) and the superficial portion of the brachioradialis (predominantly white) muscles of the rhesus monkey. The relative succinic dehydrogenase activities were quadratus femoris > soleus > sartorius > brachioradialis, the quadratus femoris having 7 times more enzyme activity than the brachioradialis. The sartorius of male rhesus monkeys had a higher enzyme activity than that of the female. Muscle samples were stained with sirius red and graded for amounts of connective tissue as follows: soleus < sartorius < brachioradialis. These histochemical results were verified by chemical analyses. The soleus, sartorius and brachioradialis from 10 other species of primates had the same relative succinic dehydrogenase activities and histochemical staining patterns as the rhesus.
STUDIES IN HISTOCHEMISTRY LXXXVI. QUANTITATIVE HISTOLOGIC DISTRIBUTION IN THE RAT ADRENAL OF ENZYMES OXIDIZING PYRIDINE NUCLEOTIDES AND EFFECTS OF ACTH
Authors: SCHOR NORBERTO A. , GLICK DAVID
Published: April 2011

For purposes of this study, micro-adaptations of established fluorometric and spectrophotometric methods were developed for quantitative measurement in microtome sections of tissue of NADPH- and NADH-dependent dichlorophenolindophenol reductases (diaphorases) and quinone reductases, and also of NADPH and NADH oxidases and NADPH-NAD transhydrogenase. Correlations of the enzyme activities with the histology of the rat adrenal gland were made, both in glands from animals injected with ACTH and from control animals given saline injections. The functional significance of the results was considered in relation to electron transport and hormone biosynthesis in the organ.
ULTRASTRUCTURAL LOCALIZATION OF ALKALINE PHOSPHATASE ACTIVITY IN THE ABSORBING CELLS OF THE DUODENUM OF MOUSE
Authors: HUGON J. , BORGERS M.
Published: April 2011

A direct lead method for the visualization of alkaline phosphatase activity at pH 9 was used in a study on the absorptive cells of the duodenum of the mouse. Lead phosphate precipitate was observed in several structures after a short incubation of glutaraldehyde-fixed frozen sections. On microvilli and in vacuoles below the terminal web, the reaction was intense and persisted even after osmic acid fixation. In the Golgi zone, in the smooth reticulum cisternae and in the dense bodies, the reaction was prominent. In these sites, the deposits were not observed after osmic acid fixation or after short incubation in media containing substrates other than β-glycerophosphate or α-naphthylphosphate.
Midwives with mobiles: A dialectical perspective on gender arising from technology introduction in rural Indonesia
Authors: Chib Arul , Chen Vivian Hsueh-Hua
Published: April 2011

Mobile phones were introduced to rural midwives in tsunami-affected Indonesia, allowing them to contact medical experts and communicate with patients. Ninety-two interviews were conducted with midwives, coordinators, doctors, and village representatives. This study applies a dialectical perspective to supplement the analytical frame of the ICT for healthcare development model (Chib et al., 2008), by addressing the multi-dimensionality of benefits and barriers. The theory of dialectical tension (Baxter and Montgomery, 1996) situates the conceptual discussion around the struggles between autonomy and subordination within gender roles, personal growth versus technological competency, and issues of economic and resource control in traditional hierarchies. We find that midwives engage in legitimization strategies, develop peer support, and focus on strategic issues to develop the capacity for agency and autonomy, despite socio-organizational barriers. Specific recommendations are offered, focusing on the resourcefulness and desire of women.
Students’ experiences of active engagement through cooperative learning activities in lectures
Authors: Cavanagh Michael
Published: April 2011

This article reports on students’ experiences of lectures which included many opportunities for active engagement through cooperative learning activities. At the end of a 13-week semester-long unit, 113 students completed a questionnaire which contained five open-ended questions focusing on the extent to which the students thought that the lecture activities helped them to learn and understand the course content and to maintain their interest and attention during the sessions. Results indicate that students valued the mix of traditional lecturing and cooperative learning tasks, particularly the variety of activities, the opportunities for small-group and whole-class discussions, the clear focus on one or two central ideas, and the authenticity of the tasks.
Using computer games for instruction: The student experience
Authors: Grimley Michael , Green Richard , Nilsen Trond , Thompson David , Tomes Russell
Published: April 2011

Computer games are fun, exciting and motivational when used as leisure pursuits. But do they have similar attributes when utilized for educational purposes? This article investigates whether learning by computer game can improve student experiences compared with a more formal lecture approach and whether computer games have potential for improving performance. Instruction was split between lectures and computer games, and student experiences were recorded using an Experience Sampling Method to capture real-time experience and feelings of flow. Results indicated that student experiences in the game mode showed increased alertness, increased feelings of being active, increased feelings of involvement and an increased perception of challenge. Flow characteristics revealed boredom during standard lectures but anxiety and flow during game modes. Finally, some evidence of improved attainment was evident. By using contemporary interactive approaches such as computer games, student learning experiences and attainment may be improved. Some practical issues of implementing games are also discussed.
Embedding academic writing instruction into subject teaching: A case study
Authors: Wingate Ursula , Andon Nick , Cogo Alessia
Published: April 2011

The benefits of embedding the teaching of writing into the curriculum have been advocated by educators and researchers. However, there is currently little evidence of embedded writing instruction in the UK’s higher education context. In this article, we present a case study in which we report the design, implementation and evaluation of an academic writing intervention with first-year undergraduate students in an applied linguistics programme. Our objectives were to try a combination of embedded instructional methods and provide an example that can be followed by lecturers across disciplines and institutions. Through the integration of in-class and online writing tasks and assessment feedback in a first-term module, we supported students’ writing development throughout the first term. We evaluated the effects of the intervention through the analysis of notes on classroom interaction, a student questionnaire and interviews, and a text analysis of students’ writing and the feedback comments over time. The evaluation findings provide insights into the feasibility and effectiveness of this approach. The embedded writing instruction was perceived as useful by both students and teachers. The assessment feedback, whilst being the most work-intensive method for the teachers, was valued most by the students and led to substantial improvements in the writing of some. These findings suggest that embedded writing instruction could be usefully applied in other higher education contexts.
Feminist participatory research in a social justice organization
Authors: Gouin Rachel R , Cocq Karen , McGavin Samantha
Published: April 2011

The authors of this article are staff members of a not-for-profit and non-hierarchical Canadian social justice organization. As members of a co-management team, they were mandated by their colleagues to lead research on the organization’s practice. In this article, they share their experience of conducting feminist participatory action research in an environment that functions by consensus. They provide an overview of the organization’s work and structure, then outline how the collective research process unfolded. The authors then discuss the synergy between the organizational structure and the research, that is, how staff members’ practice and commitment to process and consensus facilitated and strengthened the research. Finally, they share findings related to the importance of process for building alliances, learning, and collective action domestically and internationally.
ENZYMATIC DIGESTION IN THE CYTOCHEMICAL DEMONSTRATION OF GLYCOGEN
Authors: ROSA FORTUNATO , JOHNSON FRANK B.
Published: April 2011

Mounted ultrathin sections of rat fetal gastric mucosa and liver tissue, fixed in osmium tetroxide or glutaraldehyde and embedded in epoxy resin, are suitable for the cytochemical demonstration of glycogen by staining either with lead or by the periodic acid-thiosemicarbazide procedure. Enzymatic digestion with saliva or α-amylase can be performed on mounted sections of similarly treated tissues for the selective removal of glycogen prior to staining.
DIFFERENTIAL DEMONSTRATION OF MUSCLE AND HEART TYPE LACTIC DEHYDROGENASE OF RAT MUSCLE AND KIDNEY
Authors: MCMILLAN PAUL J.
Published: April 2011

A method for the histochemical demonstration of lactic dehydrogenase (LDH) is presented. This method utilizes a semipermeable membrane to minimize enzyme diffusion and high tetrazole concentration with a controlled reaction rate to minimize product diffusion. It was demonstrated that phenazine methosulfate could be used to make the reaction independent of endogenous diaphorase providing either a cytochrome oxidase inhibitor or strict anaerobiosis is employed. Differentiation of cells containing predominantly H-LDH from cells containing predominantly M-LDH was achieved by the use of 4 molal urea or pyruvate in the incubation medium. A preliminary report of the staining patterns observed in rat gastrocnemius and kidney indicates the usefulness of the technique. A new avenue to the study of the distribution of the LDH subunits is thus opened.