bluuebunny/crossref_metadata_2025 huggingface dataset:
LOCALIZATION BY PEROXIDASE-LABELED ANTIBODIES OF BOVINE CHYMOTRYPSINOGEN
Authors: SCHIFF ROBERT , KRIEG RICHARD J. , HUNTER ROBERT L.
Published: April 2011

The localization of bovine chymotrypsinogen in the pancreas was investigated by horseradish peroxidase-labeled antibodies. Chymotrysinogen was not only found in the zymogen granules of the apical portion of the acinar cells of some acini but also in the basal portion and surrounding the nuclei in other acini. Not all acinar cells exhibited the same degree of staining in a given acinus. This indicated asynchronous proenzyme production. Cytoplasmic staining was found but may have been an artifact. Gel diffusion showed a single precipitin line of identity between trypsinogen, trypsin, chymotrypsinogen, and α-, β- and γ-chymotrypsin. Because of the cross-reaction it is presumed that the antibody was recognizing both trypsinogen and chymotrypsinogen.
IMMUNOFLUORESCENT LOCALIZATION OF ORNITHINE AMINOTRANSFERASE IN RAT LIVER
Authors: BRENNAN PATRICIA C. , PERAINO C. , FRY R. J. M. , SWICK R. W.
Published: April 2011

Ornithine aminotransferase (l-ornithine:2-oxoacid aminotransferase) (EC 2.6.1.13) was localized in the livers of rats fed diets differing in protein content with an indirect fluorescent antibody technique. The ornithine aminotransferase specific fluorescence in livers from rats fed a 60% protein diet was distributed throughout the cytoplasm of hepatic cells, and numerous fluorescent cells were observed throughout the lobules. Stained hepatic cells of rats fed a standard laboratory diet (24% protein) appeared to be less numerous and the fluorescence less intense than in rats fed the 60% diet. Livers from rats fed the 0% protein diet showed no specific fluorescence. The identification of ornithine aminotransferase in situ with the fluorescent antibody technique is compatible with measurements of the activity of enzyme extracted from the livers of rats fed similar diets.
β-HEXOSAMINIDASES IN THE NERVOUS SYSTEM: THE QUANTITATIVE HISTOCHEMISTRY OF β-GLUCOSAMINIDASE AND β-GALACTOSAMINIDASE IN THE CEREBELLAR CORTEX AND SUBJACENT WHITE MATTER
Authors: SHUTER ELI R. , ROBINS ELI , FREEMAN MARY LOU , JUNGALWALA FIROZE B.
Published: April 2011

A modification of an existing fluorimetric method for the assay of β-glucosaminidase and β-galactosaminidase, based upon the hydrolysis of the corresponding 4-methylumbelliferyl glycosaminides, is described. The method is simple and convenient; its sensitivity is 3 x 10–8 M for the assay of β-glucosaminidase and 1 x 10–8 M for β-galactosaminidase. The results obtained on brain homogenates and dissected sections of cerebellar cortex and subjacent white matter of rat, rabbit and monkey were found to be similar to those obtained with the standard colorimetric procedures using p-nitrophenyl substrates. In each species, in each layer of cerebellar cortex and subjacent white matter, there were only minimal differences in the ratios of β-glucosaminidase to β-galactosaminidase activity, suggesting that a single enzyme possesses both activities. β-Glucosaminidase and β-galactosaminidase in the cerebellum were most active in the cellular granular layer, suggesting an association of β-hexosaminidase activity with neuronal cell bodies as has been found with other lysomal enzymes.
SOME NEW ASPECTS OF EFFICIENCY OF ELECTRON MICROSCOPIC AUTORADIOGRAPHY WITH TRITIUM
Authors: VRENSEN G. F. J. M. , DE GROOT D.
Published: April 2011

Quantitative aspects of submicroscopic autoradiography, using the flat substrate technique, were studied. Using Kodak D19b as developer efficiency values of about 37 for the Ilford LA emulsion and about 17 for the Gevaert-Agfa Scientia Nuc 3.07 emulsion were obtained with H3-methacrylate sections of gray interference colors. Theoretical and experimental data revealed that the high standard deviations of efficiency values obtained are caused mainly by variations in section thickness within ribbons of sections, which on account of identical interference colors were considered to be equally thick. Measurements of efficiency on sections with thickness variations between 400 and 2500 Å revealed that self-absorption is an important factor in tritium autoradiography at the electron microscopic level.
ACID PHOSPHATASE ISOENZYME IN HUMAN LEUKOCYTES IN NORMAL AND PATHOLOGIC CONDITIONS
Authors: LI C. Y. , YAM L. T. , LAM K. W.
Published: April 2011

Acid phosphatase in human leukocytes was examined in a large number of patients with a variety of hematologic diseases. In chronic lymphocytic leukemia the total leukocyte acid phosphatase activity was markedly decreased. This was due to the drastic increase of enzyme-poor leukemic lymphocytes and the concomitant decrease of enzyme-rich monocytes and neutrophils. Further examination by disc gel electrophoresis revealed that the leukocytes in this disease contained only one of the five acid phosphatase isoenzymes present in a normal leukocyte preparation. Total acid phosphatase activity was not significantly altered in other hematologic disorders, yet different ratios of the isoenzymes shown by disc gel electrophoresis were observed in Hodgkin's disease, chronic granulocytic leukemia, acute granulocytic leukemia, infectious mononucleosis and leukemic reticuloendotheliosis.
PEROXIDASE ACTIVITY IN PEROXISOMES (MICROBODIES) OF ACATALASEMIC MICE
Authors: GOLDFISCHER SIDNEY , ESSNER EDWARD
Published: April 2011

Peroxisomes (microbodies) of a strain of acatalasemic mice (Csb) are, paradoxically, more rapidly and intensely stained than those of the wild type (Csa) when incubated in a diaminobenzidine medium that demonstrates the peroxidatic activity of catalase. Biochemical studies have shown that the mutant catalase is unstable and is rapidly inactivated by heat with loss of catalatic activity. Our results indicate that the inactivated catalase of the acatalasemic mutant is an active peroxidase that oxidizes both diaminobenzidine and benzidine at neutral and alkaline pH and at 15 and 37°C and is resistant to the catalase inhibitor, 3 amino-1,2,4-triazole.
AUTORADIOGRAPHIC DETECTION OF ANTIGENS IN CELLS USING TRITIUM-LABELED ANTIBODIES
Authors: OSTROWSKI K. , BARNARD E. A. , SAWICKI W. , CHORZELSKI T. , LANGNER A. , MIKULSKI A.
Published: April 2011

Immunoglobulins specific for cellular antigens of several types were treated with 3H-acetic anhydride in conditions which were shown to introduce three to four 3H-acetyl groups per molecule, with very high retention of the antibody titers. Methods were established for application of these labeled globulins in an autoradiographic procedure. The results show that localization of cell-bound antigens can be demonstrated by this method, by silver grain concentrations at sites that correspond to those demonstrated by the same globulins labeled with a fluorescent marker. A control nonimmune globulin was reacted similarly and gave no labeling of the same tissues. The advantage of the isotopic method would be the relative quantitation that it is, in principle, capable of yielding.
HISTOCHEMICAL AND HISTOPHYSICAL INVESTIGATIONS ON THE ACETYLATION BLOCKADE OF CARBOXYLIC GROUPS OF POLYSACCHARIDES
Authors: MATERAZZI GIOVANNI , FERRETTI ELIGIO
Published: April 2011

Acetylation abolishes the metachromatic staining of certain mucins, specifically of those that stain metachromatically at pH 2.3-3.0 and in which the metachromasia is produced by oxidation with chromic acid. Mucopolysaccharides that stain metachromatically at pH 1.5 or after sulfation stain metachromatically even after acetylation. Neuraminidase treatment abolishes in some substrates (demilunes of the submandibular gland of Canis familiaris, mucous cells of the submandibular gland of Bos taurus and Ovis aries) the chromotropy and alcianophilia, whereas in others (mucous cells of the submandibular gland of Canis familiaris, demilunes of the submandibular gland of Felis catus) it causes only a weakening of these reactions, which disappear after successive acetylation. Infrared spectra of histologic sections show the appearance of a peak at 1730 cm–1 after acetylation. These studies indicate that acetylation blocks the carboxylic groups of sialic acids or of other mucins and neuraminidase-resistant sialic acid-containing substances. Infrared spectrophotometric analysis shows the presence of ester linkages upon acetylation, but does not exclude the possible formation of δ-lactones.
HIGH GLYCOGEN CONTENT OF RED AS OPPOSED TO WHITE SKELETAL MUSCLE FIBERS OF GUINEA PIGS
Authors: GILLESPIE C. A. , SIMPSON D. R. , EDGERTON V. R.
Published: April 2011

Histochemical examination of rabbit, rat, guinea pig and mouse limb skeletal muscles in our laboratory has revealed that most red muscle fibers in any given muscle stain more intensely for glycogen (periodic acid-Schiff's reaction) than white or intermediate fibers. Since most studies have concluded that white muscle has more glycogen than red muscle, this discrepancy was investigated. Eleven vastus lateralis muscles were resected from six adult male guinea pigs and cropped to yield "red," "moderate" and "white" regions. Cross-sections of each region were employed for histochemical fiber typing and the remaining pieces for biochemical glycogen analysis. These analyses have shown the red region to have significantly more glycogen (9.7 mg per g) and more red fibers (77%) than the white region (7.4 mg/g, 29%). Having essentially eliminated intermediate fibers by cropping, these findings reflect glycogen concentration in red and white muscle fibers. The existence of significant numbers of intermediate fibers usually found in red muscles may be responsible for the frequently exhibited higher glycogen content in white muscles.
MICROPROBE ANALYSIS OF PROTARGOL STAIN DEPOSITION IN TWO PROTOZOA
Authors: ZAGON IAN S. , VAVRA JIRI , STEELE IAN
Published: April 2011

Protargol-stained specimens of two protozoans, Carchesium polypinum (Ciliata, Peritrichida) and Gregarina rhyparobiae (Sporozoa, Eugregarinida), were examined by an electron microprobe analyzer in order to determine the elemental composition of the stain deposit. Limited penetration of the microprobe beam presumably revealed only the composition of the stain deposits close to the surface of the examined protozoa. The data on this first analysis of Protargol staining revealed that element detection was negligible in the unstained preparations, but the stained specimens of both protozoans revealed a mass concentration ratio of 0.4 Cu:1 Ag:6.1 Au. There was no significant difference in the element ratio between the two protozoan species belonging to different classes and stained with slightly different stain procedures. Some aspects of Protargol mechanisms are discussed along with an interpretation of data and an indication of the feasibility of further examinations.
UPTAKE OF 3H-NOREPINEPHRINE BY FLUORESCENT NERVES OF THE HEART
Authors: MASUOKA DAVID T. , PLACIDI GIAN FRANCO
Published: April 2011

The localization of uptake 5 minutes after the injection of a low dose of 3H-norepinephrine by adrenergic nerve fibers in the heart has been observed by combining autoradiography with the formaldehyde-induced histochemical fluorescence method for aromatic monoamines in the same tissue section. The distribution of 3H-label corresponds exactly to that of the fluorescent nerves. Furthermore, uptake studies in animals depleted of catecholamines by the tyrosine hydroxylase inhibitor (H 44/68), which does not block uptake, indicate that the dosage of 3H-norepinephrine used, under the conditions of the present experiment, was not in itself sufficient to produce the intense formaldehyde-induced fluorescence in nerves. It may be concluded that 3H-norepinephrine was taken up only by those nervous structures in the heart which show a specific fluorescence reaction due to the presence of endogenous catecholamine.
MURINE AMYLOID FIBRIL PROTEIN: ISOLATION, PURIFICATION AND CHARACTERIZATION
Authors: GLENNER G. G. , PAGE D. , ISERSKY C. , HARADA M. , CUATRECASAS P. , EANES E. D. , DELELLIS R. A. , BLADEN H. A. , KEISER H. R.
Published: April 2011

Murine amyloid has been produced by four different induction methods ( Mycobacterium butyricum, casein, casein plus Freund's adjuvant and endotoxin-induced mouse amyloidosis) in several strains and obtained from mice with "spontaneous" amyloidosis. The amyloid fibrils have been concentrated from spleen and liver. Electron microscopy of all of these preparations reveals the amyloid fibril to be 100 Å in width and composed of two parallel filaments, each measuring 35-40 Å in width and having the appearance of a twisted ribbon. X-ray diffraction of all preparations reveals a "backbone" spacing at 4.75 Å and a "side chain" spacing at 11 Å indicating a "β-pleated sheet" structure. Identification of the major protein component of amyloid fibril concentrates was made by combined use of sodium dodecyl sulfate polyacrylamide disc electrophoresis, labeling of the protein with 3H-tryptophan and Sepharose 4B gel filtration. Purification of the amyloid protein from spontaneous amyloidosis liver was accomplished by sequential gel filtration with 5 M guanidine in 1 N acetic acid on Sepharose 4B and Sephadex G-100 or G-75 columns. The material is a unique glycoprotein with a molecular weight of 7200, a high content of dicarboxylic and short chain amino acids, a significant amount of tryptophan and an unreactive NH2-terminal amino acid, tentatively identified as pyrrolid-2-one-5-carboxylic acid. There are no methionine, half-cystine, hydroxylysine or hydroxyproline residues. Murine amyloid protein, therefore, has striking similarities to many human amyloid protein preparations. It differs from the human proteins in the similarity of molecular weights of different preparations.
A RADIOAUTOGRAPHIC TECHNIQUE FOR WHOLE MOUNTS OF MUSCLE FIBERS
Authors: KOPRIWA BEATRIX M. , MOSS F. P.
Published: April 2011

A technique is described for the radioautography of whole mounts of muscle fibers, which are approximately 15µ in diameter and too small to be handled individually. The fixed muscle is digested with collagenase to separate the fibers; these are then stained with hematoxylin, thoroughly washed and suspended in radioautographic emulsion on microscope slides. Thus the nuclei, which are all at the surface of the fibers, are close to the surrounding emulsion. This ensures that all of the nuclei containing the radioisotope will be labeled in the radioautograph, irrespective of their position, above, below or at the side of the fibers.
LIMITATIONS OF THE PYROANTIMONATE TECHNIQUE FOR LOCALIZATION OF SODIUM IN ISOLATED CEREBRAL TISSUES
Authors: SUMI S. M. , SWANSON P. D.
Published: April 2011

Thin slices of guinea pig cerebral cortex incubated for 30-60 min with or without ouabain were analyzed for Na+, K+ and inulin contents. Other brain slices treated in a similar manner were fixed in potassium pyroantimonate-containing buffered glutaraldehyde solution and examined in the electron microscope. Slices incubated in the presence of ouabain contained considerably more Na+ in the non-inulin, presumably intracellular, space. However, precipitates of pyroantimonate which have been accepted by some to indicate the site of Na+ localization were almost entirely restricted to the extracellular space in both the control and experimental slices and appeared to reflect the method of fixation by immersion and not the movement of Na+. It is our conclusion that the lack of correlation between the chemical localization of Na+ and the pyroantimonate technique is due to the limitation of the latter method.
ADRENERGIC INNERVATION OF SPHINCTERIC AND NONSPHINCTERIC SMOOTH MUSCLE IN THE RAT INTESTINE
Authors: GILLESPIE J. S. , MAXWELL J. D.
Published: April 2011

The Falck and Hillarp fluorescence technique has been used to investigate the density of adrenergic innervation of smooth muscle at different levels in the alimentary canal of the rat. Special attention was given to potential sphincteric regions. The density of smooth muscle innervation was in general low. No special innervation was found in the region of the cardiac sphincter or at the ileocecal junction. A sharply circumscribed region of muscle with a high density of adrenergic innervation was observed in the pylorus just short of the pyloroduodenal junction and may represent a sphincteric region. In the terminal colon the anococcygeus muscle was found to form a powerful muscular loop around the colon. This muscle has a density of adrenergic innervation as high as that of the vas deferens. It can be easily isolated and forms an admirable in vitro preparation. It has not previously been described in the rat nor has its adrenergic innervation been described in any species.
AMINOACRIDINE DYES AND THE SECONDARY STRUCTURE OF DNA IN SITU
Authors: DIAZ MANUEL , HIERRO JOSE , DE DIAZ GRACIELA DEMICHELI
Published: April 2011

A new method is proposed to study the secondary structure of deoxyribonucleic acid (DNA) in situ in fixed chromatin. It is based on acriflavine staining and on differentiation with a nitrous acid solution of the fixed cytologic preparation. The presence of green fluorescence after this treatment is regarded as indicative of double stranded DNA. Experiments are described with DNA-acriflavine mixtures in solution, DNA-agar models and cytologic preparations submitted to different pretreatments. The feasibility and limitations of the method are discussed in the light of the results reported upon.
PHOSPHATASES OF EPIPHYSEAL CARTILAGE STUDIED BY ELECTRON MICROSCOPIC CYTOCHEMICAL METHODS
Authors: MATSUZAWA TATSUJI , ANDERSON H. CLARKE
Published: April 2011

The presence and distribution of alkaline phosphatase, adenosine triphosphatase (ATPase) and acid phosphatase activities in the epiphyseal plates of young mice were studied by electron microscopic cytochemical methods. Alkaline phosphatase and ATPase activities were associated with the plasma membranes of chondrocytes and with the investing membranes of matrix vesicles. These vesicles contain the earliest recognized deposits of hydroxyapatite and may promote calcification through an active process. Alkaline phosphatase and ATPase activities were greatest in the hypertrophic zone of the epiphysis, which is an area of beginning calcification. Acid phosphatase activity was demonstrable in the cytoplasm of chondrocytes in association with dense bodies which were larger than matrix vesicles and were devoid of alkaline phosphatase and ATPase activities. These cytoplasmic lysosome-like bodies were slightly more numerous in the hypertrophic zone but disappeared in the underlying zone of chondrocyte degeneration and matrix calcification. Our observations do not support a previous suggestion by others that matrix vesicles represent lysosomes. The presence of ATPase and alkaline phosphatase is compatible with an enzymatic calcium- and/or phosphate-concentrating mechanism in the matrix vesicles.
ESTERASES IN DEVELOPING AND ADULT RAT LUNG I. BIOCHEMICAL AND ELECTRON MICROSCOPIC OBSERVATIONS
Authors: O'HARE KAREN HITCHCOCK , REISS OSCAR K. , VATTER ALBERT E.
Published: April 2011

Reaction rates, kinetics, inhibition and cellular sites of nonspecific esterases in developing rat lung were studied using biochemical and cytochemical techniques. The rates of hydrolysis of p-nitrophenylthiol acetate (PNTA) and p-nitrophenylthiol butyrate (PNTB) in homogenates were correlated with ultrastructural localization of the esteratic activity. The effects of a number of esterase inhibitors such as diisopropyl phosphorofluoridate were examined in both systems. Throughout lung development there is an increase in the rates of hydrolysis of thiol esters by lung homogenates. Fetal PNTA hydrolases are most abundant in the mesoderm and are widely distributed on membranes ( i.e., Golgi complexes, nuclear envelopes, lipid droplets). The major sites of endodermal PNTA hydrolysis are lamellar bodies and their proposed precursors. PNTB hydrolases are more limited in distribution. Regardless of substrate used, adult esteratic activity is most prominent in lamellar bodies of type II epithelial cells. The possible relationship of esterase activity, lamellar bodies and surfactant is discussed.
VISUALIZATION IN FREEZE-DRIED TISSUE SECTIONS OF STRUCTURES TO BE STUDIED BY QUANTITATIVE HISTOCHEMISTRY
Authors: HELLMAN BO , LERNMARK ÅKE
Published: April 2011

The limiting factor for quantitative histochemistry employing freeze-dried tissue sections is the ability to identify morphologic structures. Satisfactory staining of freeze-dried tissue specimens intended for further microanalysis was achieved by using isopentane solutions of free dye bases. The practical details of this rapid histologic control are outlined and its usefulness is illustrated by measurements of fructose 1,6-diphosphate and insulin in microdissected specimens of pancreatic β cells.
AN OSMIOPHILIC DISTYRYL DITETRAZOLIUM SALT (DS-NBT) FOR ULTRASTRUCTURAL DEHYDROGENASE ACTIVITY
Authors: SELIGMAN ARNOLD M. , NIR IZHAK , PLAPINGER ROBERT E.
Published: April 2011

The preparation of a new osmiophilic ditetrazolium salt (DS-NBT) containing two styryl groups, but retaining the essential structural features of nitro-BT, is given. DS-NBT may be used to demonstrate succinic dehydrogenase activity (SDH) faster than with nitro-BT and fully as rapidly as with TC-NBT. DS-NBT is more stable than TC-NBT and it is more reliably and more readily osmicated than TC-NBT. The ultrastructural sites of SDH and NADH2 diaphorase activity are similar to those shown for cytochrome oxidase activity with diaminobenzidine in rat heart. The sites of lactic dehydrogenase activity are found in the sarcoplasmic reticulum and mitochondria of rat skeletal muscle fixed in formaldehyde for 3 hr. Tissue penetration of the doubly charged ditetrazolium salt (DS-NBT) was facilitated by 40-sec sonication and by 5% dimethyl sulfoxide. Illustrative examples and controls are included.
HIGH RESOLUTION AUTORADIOGRAPHY WITH STRIPPING FILM
Authors: WILLIAMSON JOSEPH R. , VAN DEN BOSCH HANK
Published: April 2011

A simple method is described for the preparation and use of stripping film of high quality for electron microscopic applications of autoradiography. The advantages of the method are: ( a) uniform monolayers of emulsion are easily prepared with an inexpensive mechanical device and applied to thin sections on conventional grids; ( b) both sides of grids can be coated with emulsion, permitting a potential doubling of sensitivity with, theoretically, no loss of resolution; and ( c) large numbers of grids can be processed simultaneously, assuring identical conditions of development.
LOCALIZATION AND ONSET OF AMYLASE ACTIVITY IN MOUSE SALIVARY GLANDS DETERMINED BY A SUBSTRATE FILM METHOD
Authors: SMITH RICHARD J. , FROMMER JACK , SCHIFF ROBERT
Published: April 2011

A starch substrate film technique was used to determine the localization and onset of amylase activity in the major salivary glands of mice. The earliest activity occurred in the parotid gland of males and females at 8 days of age. The intensity of the reaction rapidly increased so that by day 16 the entire parotid gland was amylase-positive. The submandibular gland exhibited the sexual dimorphism that has been characterized for the adult gland. Amylase activity was observed in the male at 24 days of age, shortly after the first morphologic differences between the submandibular glands of males and females became apparent. The female showed positive results at 36 days of age. Amylase activity in the submandibular glands was localized to the convoluted tubules. As the gland developed, the amylase activity increased in correlation with the increasing concentration of these tubules. The sublingual gland was always amylase-negative.
HISTOCHEMICAL COMPARISON OF FROG AND RAT MAST CELLS
Authors: CHIU HERBERT , LAGUNOFF DAVID
Published: April 2011

Histochemical procedures were used to compare the constituents of frog and rat mast cells. On the basis of staining with acidified toluidine blue and the Alcian Blue-safranin combination, a substance characterized as heparin in rat cells was identified in frog mast cells. In contrast to their presence in rat cells, no histamine, serotonin or catecholamines could be observed in frog mast cells with the o-phthalaldehyde and paraformaldehyde reactions. A search for activities of two proteolytic enzymes, chymotrypsin-like and trypsin-like esterases, in frog mast cells was unsuccessful. β-Glucuronidase, N-acetyl-β-d-glucosaminidase and nonspecific esterase activities were demonstrated in both frog and rat mast cells; acid phosphatase activity was detected in frog but not rat mast cells.
HISTOCHEMICAL LOCALIZATION OF BRAIN SUCCINIC SEMIALDEHYDE DEHYDROGENASE-A γ-AMINOBUTYRIC ACID DEGRADATIVE ENZYME
Authors: SIMS K. L. , WEITSEN H. A. , BLOOM F. E.
Published: April 2011

Succinic semialdehyde dehydrogenase is the final enzyme in the degradative pathway of γ-aminobutyric acid, a postulated inhibitory synaptic transmitter. This study reports a specific, sensitive technique for the localization of succinic semialdehyde dehydrogenase in rat brain. The technique utilizes the direct reduction of nitroblue tetrazolium by reduced pyridine nucleotide at an alkaline pH, a mechanism of formazan production substantiated by biochemical studies. The succinic semialdehyde dehydrogenase histochemical reaction is completely inhibited by p-hydroxybenzaldehyde and m-hydroxybenzaldehyde, compounds which have no effect on succinic acid dehydrogenase and reduced diphosphopyridine nucleotide diaphorase. Conversely, succinic acid dehydrogenase and reduced diphosphopyridine nucleotide diaphorase are inhibited by malonate and nordihydroguaiaretic acid, respectively, substances which have no effect on succinic semialdehyde dehydrogenase. The distribution and intensity of staining for succinic semialdehyde dehydrogenase are different from that seen following the succinic acid dehydrogenase and reduced diphosphopyridine nucleotide diaphorase reactions and can be localized to groups of neurons and their cell processes, e.g., the Purkinje cell dendritic tree, and in specific regions of neuropil.
LACTIC ACID DEHYDROGENASE ISOENZYMES OF RAT SOLEUS MUSCLE FIBERS AS DEMONSTRATED BY HISTOCHEMICAL STAINING AND ELECTROPHORESIS
Authors: MCMILLAN PAUL J. , WITTUM ROGER L.
Published: April 2011

A histochemical stain for heart type lactic dehydrogenase (LDH) has been correlated with the electrophoretic isoenzyme pattern. The rat soleus was chosen for this study since a minority of its fibers contain LDH which is inhibited by urea in the histochemical system. This minority group has attracted attention because they stain intensely for both phosphorylase and succinic dehydrogenase. Electrophoresis of LDH from fibers dissected from freeze-dried sections revealed that the bulk of the soleus fibers contain primarily isoenzymes I and II with a trace amount of III and practically none of IV and V. The isoenzyme pattern obtained from the minority group of fibers stained most intensely for isoenzyme III. Isoenzyme II gave a moderate reaction while isoenzymes I and IV reacted weakly and isoenzyme V was not detectable. Taken with other observations these results indicate that in the histochemical system containing 4 molal urea a strong reaction in rat tissues is evidence of a preponderance of isoenzymes I and perhaps II.
SUBPOPULATIONS OF BLOOD LYMPHOCYTES DEMONSTRATED BY QUANTITATIVE CYTOCHEMISTRY
Authors: GIACOMELLI FILIBERTO , WIENER JOSEPH , POMERANZ JOANNA V. , LOUD ALDEN V. , KRUSKAL JOSEPH B.
Published: April 2011

The cell coats of glutaraldehyde-fixed blood lymphocytes from guinea pigs have been stained with Thorotrast and examined by electron microscopy. The lymphocytes have been classified into three groups with respect to the concentration and distribution of colloidal particles over their surfaces. The average number of particles per micron length of cell surface membrane, the particle density, has then been measured for each class of cells. These values are distributed into two clusters separated by a gap. A new statistical test for bimodality has been devised to evaluate the significance of such a gap in terms of its "dip intensity." The results of this analysis demonstrate the existence of at least two distinct populations of blood lymphocytes, indistinguishable by other morphologic criteria.
COMPARISON BETWEEN 3,3'-DIAMINOBENZIDINE AND AUTO-OXIDIZED 3,3'-DIAMINOBENZIDINE IN THE CYTOCHEMICAL DEMONSTRATION OF OXIDATIVE ENZYMES
Authors: HIRAI KEI-ICHI
Published: April 2011

The interactions between auto-oxidized 3,3'-diaminobenzidine (DAB oxide) and mitochondria and peroxisomes were studied electron microscopically. Mitochondria were visualized in heart and liver thoroughly fixed with glutaraldehyde or formaldehyde after incubation in auto-oxidized DAB medium. Dense deposits were seen along the inner membranes of the mitochondria probably due to coupling of DAB oxide with cytochrome c and possibly with cytochrome a, but not with b type cytochromes. This reaction in formol-fixed tissues was stable to heat for 20 min at 80°C. Dense deposits of conjugated catalase of hepatic peroxisomes occurred in the matrices, and the reaction was sensitive to 3-amino-1,2,4-triazole. Heating for 30 min at 120°C of formol-fixed liver had no effect on this reaction while enzyme activities of peroxisome catalase were destroyed. With incubation in fresh DAB medium for 3 hr no staining of cytoplasmic organelles was visualized. These reactions of DAB oxide were nonenzymatic and resulted from physical bindings. The possible chemical states of DAB oxide are discussed. Some free radicals were detected in the precipitates, but no information has been obtained from aqueous solutions. The auto-oxidation of DAB is mainly a photo-oxidation.
LOCALIZATION OF SPECIFIC PHOSPHATASE ACTIVITIES IN CANINE CORONARY BLOOD VESSELS AND HEART MUSCLE
Authors: BORGERS M. , SCHAPER J. , SCHAPER W.
Published: April 2011

The localization of nonspecific alkaline phosphatase (AIPase), acid phosphatase (ACPase), nucleoside phosphatases (ATP-, ADP- and AMP-splitting enzymes), thiamine pyrophosphatase (TPPase) and glucose 6-phosphatase (G-6-Pase) activities was studied in coronary blood vessels and myocardium of the canine left ventricle. Except for AIPase and 5'-AMPase, the subcellular distribution of enzyme activities was essentially the same in arterial and venous vessels of different sizes, although enzyme activity varied in degree. Plasma membranes and pinocytic vesicles of intimal, medial and adventitial cells were reactive toward ATP, ADP and TPP. 5'-AMPase activity was seen at the cell membrane of adventitial or perivascular mesenchymal cells only. AIPase was confined to the endothelium of resistance vessels. Lysosomes and some Golgi complex showed moderate ACPase activity. Some Golgi cisternae and vesicles stained strongly with TPP as substrate. The endoplasmic reticulum and the nuclear envelope were constantly positive for G-6-Pase in every cell constituting the vascular wall. In the myocardial cell, ATPase activity was confined to the lateral elements of diads and triads, the subsarcolemmal cisternae and the mitochondrial matrix. Activity toward TPP and ADP was only seen in the Golgi apparatus. The few lysosomes present in myocardial cells possessed ACPase activity. G-6-Pase activity was uniformly distributed in the sarcoplasmic reticulum, the nuclear membranes, the subsarcolemmal cisterns and lateral elements of diads and triads.
A Treatment Algorithm for Primary Patellar Dislocations
Authors: Jain Neel P. , Khan Najeeb , Fithian Donald C.
Published: April 2011

Context:

Primary patellar dislocation continues to be a common problem facing clinicians today. These injuries are associated with significant morbidity and a substantial recurrence rate. Myriad operative and nonoperative options have been described to treat these injuries, although the evidence-based literature is sparse.

Evidence Acquisition:

PubMed was searched from 1970-2010 to identify publications on patellar dislocations, including clinical presentation, natural history, radiographic workup, and treatment.

Results:

The initial workup of a patella dislocation includes a history, physical examination, and radiographs. If there is evidence of a displaced osteochondral fragment or hemarthrosis, then magnetic resonance imaging should be obtained. The treatment of first-time patella dislocators has been controversial, and no study has demonstrated a clear benefit to early operative intervention.

Conclusion:

First-time patellar dislocations should be treated conservatively unless there is evidence of a displaced osteochondral fragment.
HISTOCHEMISTRY OF THE ENTEROCHROMAFFIN SUBSTANCE
Authors: GOMORI G.
Published: April 2011

The histochemical reactions of resorcinol and of 5-hydroxytryptamine are very similar. It cannot be decided, on the basis of histochemical tests at present available, whether the reactions of human EC granules are due to a derivative of resorcinol or of 5-hydroxytryptamine or of some other, unknown, nucleus.
A SIMPLE NEW MICROTOME FOR ULTRATHIN SECTIONING
Authors: HODGE ALAN J. , HUXLEY HUGH E. , SPIRO DAVID
Published: April 2011

A simple easily-constructed microtome capable of cutting thin sections of the order of 300 Å in thickness for electron microscope study is described. Design criteria for such microtomes, as well as some of the problems in thin sectioning are discussed.
THE CHEMICAL BASIS OF THE PERIODIC ACID SCHIFF REACTION OF COLLAGEN FIBERS WITH REFERENCE TO PERIODATE CONSUMPTION BY COLLAGEN AND BY INSULIN
Authors: BANGLE RAYMOND , ALFORD WILLIAM C.
Published: April 2011

1. Oxidation of collagen fibers in tissue sections or of Standard Hide Powder (hide collagen) in the testtube by an acid solution of periodate (pH 1.9) resulted in a product which gave a positive (pink to red) Schiff reaction. Oxidation by an alkaline solution of periodate (pH 8.2) gave an equivocal (faint pink) reaction.

2. The reaction between the insoluble oxidation product in collagen and the Schiff reagent was blocked completely, both in tissue sections and in the testtube, by an aqueous solution of aniline chloride, phenylhydrazine, or dimedone at room temperature, but not by the aqueous solvent alone, indicating that free insoluble aldehyde groups were produced as the result of acid periodate oxidation.

3. The acid periodate Schiff reaction of collagen fibers was not due to the presence of lipid or glycogen and did not depend upon chemical fixation of the tissue.

4. Periodate cleavage oxidation of the ε-amino-δ-hydroxy grouping of hydroxylysine in collagen did not seem to play a significant role in the acid periodate Schiff reaction.

5. Periodate cleavage oxidation of the serine and threonine constituents of collagen probably does not occur.

6. The largest fraction of carbohydrate in the dry hide powder was of nonglucosamine polysaccharide, calculated as 0.55% (expressed as glycogen). The quantity of hexosamine and uronic acids in the dry, defatted hide powder was 0.047% and less than 0.039% respectively. By indirect evidence it is deduced that the nonglucosamine polysaccharide complex conjugated with collagen most likely was responsible for the acid periodate Schiff reaction.

7. A large fraction of the periodate consumed by hide collagen could not be accounted for by oxidation of the carbohydrate content, by oxidation of vicinal hydroxy amino groups in the protein, or by nonspecific adsorption of –IO4. Evidence based upon the experiments with purified insulin indicates that periodate may be consumed by protein without the production of any aldehydes or free ammonia. This is suggested as the fate of periodate not otherwise accounted for in oxidized hide collagen.
STUDIES IN HISTOCHEMISTRY: XXVIII. THE QUANTITATIVE HISTOLOGICAL DISTRIBUTION OF ASCORBIC ACID IN THE ADRENAL GLAND OF THE RAT AND MONKEY
Authors: BAHN ROBERT C. , GLICK DAVID
Published: April 2011

The quantitative histological distribution of ascorbic acid in the adrenal gland of the normal M. rhesus and M. cynomolgus monkey and the albino rat was studied. Although the concentration of ascorbic acid is 4-5 times greater in the gland of the rat than it is in that of the monkey, the same qualitative distribution of ascorbic acid was found in both. The highest concentrations of ascorbic acid were demonstrated in the outer fascicular and in the reticular zones. Only very small concentrations of dehydroascorbic and diketogulonic acids were found in any of the histological regions in the adrenal of the rat.
HISTOCHEMICAL DEMONSTRATION OF ACID PHOSPHATASE BY AN IMPROVED AZO-DYE METHOD
Authors: BURTON J. F.
Published: April 2011

Histochemical demonstration of acid phosphatase by an azo-coupling procedure has been investigated and conditions established whereby acceptable histological localisations without evidence of diffusion artefact may be secured. These include catalysis of the coupling reaction and retarding of the rate of enzymic liberation of free naphthol. More rapid coupling is achieved in a medium of pH 5.5-pH 6.0 by the use of a high concentration of diazonium salt, by incubation at 37°C. and by the presence of a quinoline derivative, percain. Liberation of free naphthol occurs relatively slowly at the non-optimal pH range employed and is further restricted by lowering of substrate concentration; in this manner local overloading of the coupling system at sites of high enzyme activity is avoided, with consequent avoidance of diffusion of free naphthol prior to coupling. The dye produced is of a highly contrasting color and its insolubility in ethanol and xylol enables the mounting of tissues in synthetic media. The procedure is applicable to both Formalin and acetone fixed tissues, either frozen sections or paraffin ribbons, adequate pigment deposition being achieved after incubation periods of 15-120 minutes.
THE ROLE OF MICROSOMES IN THE INCORPORATION OF AMINO ACIDS INTO PROTEINS
Authors: KELLER ELIZABETH B. , ZAMECNIK PAUL C. , LOFTFIELD ROBERT B.
Published: April 2011

These studies are an extension of previous findings of a relationship between ribose nucleic acid and protein synthesis (Brachet, Caspersson, Borsook, Keller, Siekevitz, Mirsky). In vivo experiments have been described which show that 70 per cent or more of the initial incorporation of leucine-C14 or valine-C14 in the liver occurs in the protein of the microsome fraction, which contains most of the cytoplasmic ribose nucleic acid. A straight line rate of incorporation of valine-C14 into microsome protein has been found during the first few minutes after intravenous injection of the labeled amino acid. The uptake of leucine or valine into microsome protein in these experiments is approximately 2 per cent per hour.

Experiments on a cell-free system have also been reported, showing that the microsome fraction and the "soluble fraction" of the liver cell are both essential for incorporation. This incorporation occurs almost entirely into the protein of the microsome fraction.
HISTOCHEMICAL OBSERVATIONS ON RODS AND CONES IN RETINAS OF VERTEBRATES
Authors: SIDMAN RICHARD L. , WISLOCKI GEORGE B.
Published: April 2011

Retinas containing varied proportions of rods and cones, including those of frog, Amphiuma, turtle, chick, kitten and monkey, have been studied by histochemical means.

In all visual cells ribonucleic acid occurs exclusively in the myoids. These structures also sometimes contain glycogen. The paraboloids, whenever present, contain glycogen and nothing else. The ellipsoids are colored by mitochondrial stains and by sudan black B, and are rich in a mildly acidophilic protein. The outer segments of the rods and cones contain an acidophilic protein and lipids which are characterized by sudanophilia, the Ashbel-Seligman carbonyl reaction, and autofluorescence. The outer segments of the rods also contain a complex carbohydrate and react strongly for protein-bound sulfhydryl groups. The carbohydrate and the sudanophilic lipid are present in the outer segments of immature photoreceptor cells before the onset of vision and are considered to be structural components; the sulfhydryl reaction, on the other hand, occurs only in outer segments of mature rods and may represent opsin. Retinene, the carotenoid component of the visual pigments, is a fatty aldehyde which we have not identified histochemically. Inconstant, faint metachromasia appears between the outer segments of the photoreceptor cells and the processes of the pigmented epithelial cells. The basal cytoplasm of the pigmented epithelium contains acidophilic protein, sulfhydryl groups, ribonucleic acid, lipids demonstrable by their sudanophilia, carbonyl staining, and autofluorescence, and stains for the enzymes acetylcholinesterase and lipase.

The ellipsoids and outer segments of the visual cells, and the basal cytoplasm of the pigmented epithelium show similar histochemical reactions, whereas the myoids stain quite differently. Differences in histochemical properties of rods and cones in certain species are described.
TRANSPORT ADENOSINE TRIPHOSPHATASE CYTOCHEMISTRY II. CYTOCHEMICAL LOCALIZATION OF OUABAIN-SENSITIVE, POTASSIUM-DEPENDENT PHOSPHATASE ACTIVITY IN THE SECRETORY EPITHELIUM OF THE AVIAN SALT GLAND
Authors: ERNST STEPHEN A.
Published: April 2011

A cytochemical procedure is described for the ultrastructural localization of K-dependent, ouabain-sensitive nitrophenyl phosphatase activity in avian salt gland. Cryostat sections (50 µ) of paraformaldehyde-fixed tissue were incubated in a kinetically defined medium containing: 5 mM p-nitrophenyl phosphate, 10 mM MgCl2, 10 mM KCl, 100 mM Tris-HCl buffer (pH 8.5 or 9.0) and 20 mM SrCl2 to precipitate hydrolyzed phosphate. After incubation at room temperature, the sections were treated with Pb(NO3)2 to convert SrPi to PbPi precipitates for visualization in the electron microscope. Reaction product was localized on the cytoplasmic side of the secretory cell lateral and basal plasma membranes. Little, if any, reaction product was associated with the apical surfaces of the secretory cells or with endothelial surfaces of capillaries. Appropriate control experiments indicated that deposition of reaction product was dependent on Mg and K and was sensitive to ouabain. Furthermore, nonenzymatic hydrolysis of nitrophenyl phosphate did not occur in the medium, and deposition of artifactually produced precipitates did not resemble deposition of enzymatically produced precipitates. The relationship of this localization to transport adenosine triphosphatase cytochemistry is discussed, and the physiologic implications of the localization for tracing the route of active Na transport in the salt gland are considered.
CYTOCHEMICAL LOCALIZATION OF ADENYL CYCLASE ACTIVITY IN RAT ISLETS OF LANGERHANS
Authors: HOWELL S. L. , WHITFIELD MARGARET
Published: April 2011

A cytochemical method has been used to investigate the localization of adenyl cyclase activity in A and B cells of isolated rat islets of Langerhans. Adenosine triphosphate was initially utilized as substrate, the pyrophosphate liberated being precipitated by lead ions at its site of production. The specificity of the method was increased by the use of adenylyl-imidodiphosphate as an alternative substrate; this adenosine triphosphate analogue was not hydrolyzed by adenosine triphosphatase but provided an effective substrate for adenyl cyclase. Adenyl cyclase activity, which was found to retain its glucagon and fluoride sensitivity in glutaraldehyde-fixed tissue, was found exclusively and almost uniformly in the plasma membranes of A and B cells. Storage granule membrane, incorporated into the plasma membrane during secretion of the granule content by exocytosis, appeared to be devoid of adenyl cyclase activity.
A CYTOCHEMICAL EVALUATION OF PYROANTIMONATE BINDING TO THE PLASMALEMMA OF BLOOD AND BONE MARROW CELLS AND ITS RELATIONSHIP TO CELLULAR MATURATION
Authors: ACKERMAN G. ADOLPH , CLARK MICHAEL A.
Published: April 2011

Normal human blood and bone marrow cells exposed to the pyroantimonate reaction exhibit a selective binding of pyroantimonate to sites associated with the plasmalemma of neutrophilic leukocytes, erythrocytes and certain of their precursors. Chemical distinctions exist between the plasmalemma of different types of blood and bone marrow cells, and major changes in the cationic binding ability of neutrophilic and erythrocytic cells can be correlated with different phases in their differentiation. Plasmalemmal pyroantimonate-osmium-reactive cation is bound with glycoprotein in the erythrocytic elements and to both phospholipid and glycoprotein moieties in neutrophilic cells. The cellular distribution, membrane localization and degree of membrane reactivity obtained with the pyroantimonate-osmium reaction is distinct from that obtained with Thorotrast and ruthenium red.
STUDIES ON MICROPEROXISOMES II. A CYTOCHEMICAL METHOD FOR LIGHT AND ELECTRON MICROSCOPY
Authors: NOVIKOFF ALEX B. , NOVIKOFF PHYLLIS M. , DAVIS CLEVELAND , QUINTANA NELSON
Published: April 2011

A modification of the Novikoff-Goldfischer alkaline 3,3'-diaminobenzidine medium for visualizing peroxisomes is described. It makes possible light microscopic as well as electron microscopic studies of a recently described class of peroxisomes, the microperoxisomes. Potassium cyanide (5 x 10–3 M) is included in the medium to inhibit mitochondrial staining, the pH is 9.7 and there is a high concentration of H2O2 (0.05%). Two cell types have been chosen to illustrate the advantages of the new procedure for demonstrating the microperoxisomes: the absorptive cells in the human jejunum and the distal tubule cells in the guinea pig kidney. Suggestive relations of microperoxisomes and lipid are described in the human jejunum. The microperoxisomes are strategically located between smooth endoplasmic reticulum that radiates toward the organelles and contains lipid droplets and "central domains" of highly specialized endoplasmic reticulum which do not show the lipid droplets. The microperoxisomes are also present at the periphery of large lipid-like drops. In the guinea pig kidney tubule there is a striking difference between the thick limb of Henle and distal tubule. The distal tubule has a population of cells with large numbers of microperoxisomes readily visible by light microscopy; these cells are not present in the thick limb of Henle. Other differences between the two are also described.
IDENTIFICATION OF PEROXISOMES IN THE RAT ADRENAL CORTEX
Authors: BEARD MARGARET E.
Published: April 2011

Organelles with the ultrastructure and cytochemical characteristics of peroxisomes (microbodies) have been identified in cells of the zona fasciculata and zona reticularis of the rat adrenal cortex. These peroxisomes appear as small, elliptical to spherical or branched structures, enclosed by a single membrane and composed of a moderately electron-dense matrix. They do not possess a nucleoid or core of the type found in peroxisomes of liver and kidney. These organelles show a strongly positive staining reaction with the diaminobenzidine technique for peroxidatic activity of catalase. This staining is inhibited by aminotriazole. In cytochemical preparations revealing acid phosphatase activity, lysosomes are strongly stained and peroxisomes are free of reaction product.
A HISTOCHEMICAL INVESTIGATION OF INTRAFUSAL FIBERS IN TORTOISE MUSCLE SPINDLES
Authors: CROWE ALAN , RAGAB ABDUL H.M.F.
Published: April 2011

Histochemical investigations upon intrafusal muscle fibers of spindles in the extensor digitorum brevis 1 muscle of the tortoise have been carried out. The localization of phosphorylase, succinic dehydrogenase and adenosine activities together with the demonstration of lipids by the propylene glycol-Sudan method all failed to produce results which could be used to categorize the intrafusal fibers into more than one type. From these results and from previous histologic investigations it is suggested that the tortoise muscle spindle contains just one kind of intrafusal muscle fiber.
THE IMMUNOCYTOCHEMICAL DEMONSTRATION OF CATHEPSIN D
Authors: POOLE A. R. , DINGLE J. T. , BARRETT A. J.
Published: April 2011

Cathepsin D has been demonstrated for the first time using an immunocytochemical approach and fluorescence microscopy. It has been localized in chick epithelium-like cells and in cells of synovia from joints of rabbits with an experimental arthritis. The enzyme is associated with lysosome-like organelles: it also has a diffuse cytoplasmic distribution in these cells.