bluuebunny/crossref_metadata_2025 huggingface dataset:
CHOLINESTERASES AND NONSPECIFIC ESTERASES OF DEVELOPING AND ADULT (NORMAL AND ATROPHIC) RAT GASTROCNEMIUS 1. CHEMICAL ASSAY AND ELECTROPHORESIS
Authors: BARRON KEVIN D. , ORDINARIO A. T. , BERNSOHN J. , HESS A. R. , HEDRICK M. T.
Published: April 2011

Cholinesterases and nonspecific esterases of normal and atrophic (denervated and tenotomized) adult rat gastrocnemius were assayed, the latter 5-45 days after operation. Acetylcholinesterase activity per whole muscle was about 25% of normal 5 days after denervation and, in permanently denervated animals, showed little change thereafter. Tenotomy caused a transient 27% fall. Butyrylcholinesterase was unaffected. Nonspecific esterase activity of denervated gastrocnemius (substrate, α-naphthyl acetate) was normal 30 days after reversible sciatic injury and 50% of normal 45 days after irreversible denervation. Wet weight and total protein fell in approximate parallel. Results of assay and electrophoresis (zymograms) showed that muscle esterase was mainly of B type, although A and C type acetylesterases accounted for about 10% of the free or soluble fraction. An increase in the free esterase activity of the homogenate and an absolute increase in A type esterase accompanied denervation and disuse. Bound esterase was 80% released by Triton X-100 and was entirely B type. Zymograms of neonatal muscle were distinguished by the prominence of A-esterases and the presence of an acetylcholinesterase which was not encountered in adults. Over-all, despite some points of similarity, zymograms of neonatal and atrophic adult muscles were distinctly different.
SOME METHODS OF ELECTRON MICROSCOPIC VISUALIZATION OF LIPOPROTEINS IN PLASMA AND CHYLE
Authors: JONES ALBERT L. , PRICE J. MICHAEL
Published: April 2011

Using isolated rat chylomicrons and human hyperlipemic plasma as experimental systems, we have described various methods of preparing lipoproteins for electron microscopic observation. Of these methods three have been found to be particularly useful and are described in detail. They are: (1) long osmium fixation followed by ultracentrifugation of the fixed particles to form a pellet for subsequent embedding and thin sectioning, (2) brief osmium fixation followed by shadow casting of the particles and (3) osmium vapor fixation and direct electron microscopic visualization of the lipoproteins. Shadow casting appears to be the most rapid and reliable means of evaluating the particle size distribution of human plasma lipoprotein. Because of its rapidity, this procedure may be of diagnostic value in clinical research laboratories.
CONDITIONS INFLUENCING THE INTENSITY OF THE FEULGEN REACTION
Authors: DEITCH ARLINE D. , WAGNER DIETER , RICHART RALPH M.
Published: April 2011

Hydrolysis in 5 N hydrochloric acid at room temperature is preferable to the conventional Feulgen hydrolysis in 1 N HCl at 60°C. The intensities achieved are less critically dependent on time and temperature and the maximum Feulgen values obtained may be significantly higher than after the latter procedure. However, the shape of the 5 N hydrolysis curve is dependent on the fixative and preparative procedures used and it may vary from experiment to experiment. The small lymphocyte, which has previously been found by several workers to bind about 10% less Feulgen dye than other diploid cells, was found to have a similar lower Feulgen intensity after 5 N HCl hydrolysis at room temperature. Isolating leukocyte nuclei or extracting their acid-soluble nuclear proteins failed to cause a significant elevation in the amount of Feulgen dye found.
HISTOLOGIC AND IMMUNOFLUORESCENT STUDIES ON THE SITE OF ORIGIN OF GLUCAGON IN MAMMALIAN PANCREAS
Authors: OKADA NORIO , TAKAKI RYOSABURO , KITAGAWA MASAYASU
Published: April 2011

The present study was designed to investigate the site of origin of glucagon in the adult rabbit pancreas. Tissue fragments, fixed with Carnoy's solution and embedded in paraffin, were sectioned 4 µ in thickness. In a section stained with Gomori's chromium hematoxylin-phloxine, several islets were marked and photographed for comparison with immunofluorescent staining. The same section was then stained by an indirect immunofluorescent method using antiglucagon serum to identify glucagon-containing cells after transfer through xylene, decreasing concentrations of alcohol and buffered saline in succession. The islets previously marked were again observed and photographed under the fluorescent microscope. The specificity of the fluorescent staining was confirmed by the block test, absorption test and others. Good agreement between histologic and immunofluorescent staining of identical sections indicated the α cell origin of glucagon in the rabbit pancreas.
STUDIES ON CROSS-REACTIVITY OF ANTIRIBONUCLEASE ANTISERA WITH HETEROLOGOUS MAMMALIAN RIBONUCLEASES
Authors: MORIKAWA SHIGERU , YAMAMURA MASAO , HARADA TAKAYUKI , HAMASHIMA YOSHIHIRO
Published: April 2011

The cross-reactivity of antibovine alkaline ribonuclease (RNase) rabbit antiserum or antibovine acid RNase rabbit antiserum with heterologous alkaline or acid RNase was studied immunochemically and immunohistochemically. Immunochemical cross-reactivity was determined quantitatively by the inhibitory effect of antiserum on enzyme activity of appropriate RNase from various sources, human, rat, etc. Antibovine alkaline RNase antiserum strongly inhibited the alkaline RNase activity of human, rabbit, rat, mouse or guinea pig liver. On the other hand, antibovine acid RNase antiserum inhibited the acid RNase activity of human, rat or mouse spleen only moderately. Sections from human, rabbit, rat, mouse or guinea pig pancreas and spleen were stained with either fluorescein-labeled antialkaline RNase or antiacid RNase antibody. Fluorescent staining results confirmed the results of the inhibitory effect, and the patterns were essentially identical with those observed in bovine pancreas or spleen, except in the islets of Langerhans in the rabbit pancreas, in which some substances in the cytoplasm of the α cells were observed to react with either rabbit γ-globulin or guinea pig γ-globulin; this reaction was diminished by the procedure of paraffin embedding.
A NINHYDRIN HISTOCHEMICAL TEST SPECIFIC FOR HAIR KERATIN
Authors: HOLMES ELIZABETH J.
Published: April 2011

The reaction of ninhvdrin with α-amino groups of amino acids and proteins results in deamination of the amino acid or protein to form a nitrogen-containing blue dye complex. This molecule is small and highly diffusible, a property which was utilized in the development of a ninhvdrin histochemical test specific for hair keratin. Using a silicone rubber cement. which is resistant to boiling solutions, for the attachment of tissue sections to slides, the sections could be boiled in a ninhydrin solution. The ninhydrin reaction product is lost to the surrounding medium in the case of all tissues except hair keratin. Hair keratin is stained blue. No other keratin. including epidermal keratin or inner root sheath keratin. stains by this method. Although the exact chemistry of the reaction is unknown. evidence is cited for the probability that only a certain molecular arrangement of cystine in a polypeptide chain found almost exclusively in hair keratin can form an attachment site for the dye molecules.
THE LOCALIZATION OF ARYL SULFATASE ACTIVITY IN THYROID FOLLICLE CELLS
Authors: SELJELID ROLF , HELMINEN HEIKKI J.
Published: April 2011

The localization of aryl sulfatase activity in thyroid follicle cells was studied with cytochemical techniques both on the light and the electron microscopic level. Twenty male rats received thyroxine to suppress the thyroid activity. To 10 of these animals, thyrotropic hormone (TSH) was given 15-140 min before sacrifice. In both experimental groups, there was dense precipitation indicating aryl sulfatase activity in the cytosomes. In the TSH-stimulated group colloid droplets were observed. These did not show aryl sulfatase activity, not even after prolonged incubation. The number of enzyme-containing bodies in the cytoplasm seemed to decrease after TSH-stimulation, probably because of fusion of colloid droplets with preexisting cytosomes. The lack of enzyme activity in colloid droplets is discussed and some possible explanations are offered.
p-(ACETOXYMERCURIC) ANILINE DIAZOTATE, A REAGENT FOR VISUALIZING THE NAPHTHOL AS-BI PRODUCT OF ACID HYDROLASE ACTION AT THE LEVEL OF THE LIGHT AND ELECTRON MICROSCOPE
Authors: SMITH R. E. , FISHMAN WILLIAM H.
Published: April 2011

Diazotized acetoxymercuric aniline in a postcoupling procedure for β-glucuronidase and acid phosphatase using naphthol AS-BI β-d-glucosiduronic acid and naphthol AS-BI-phosphoric acid respectively as substrates has yielded excellent 2-µ sections from Araldite-embedded specimens for light microscopy and correlating electron micrographs for both enzymes. The reaction of the red pigment product with thiocarbohydrazide is believed to establish a mercury-sulfur linkage which is apparently responsible for the pigment's surviving ethanol dehydration and epoxy embedding. Osmication of the tissue before dehydration appears to increase electron density of the product apparently by either chelation to the pigment product, by reduction of osmium tetroxide or by its decomposition with mercuric sulfide. The two acid hydrolases have been demonstrated within the cisternae of the Golgi and the endoplasmic reticulum and within the nuclear envelope of parenchymal cells of mouse and rat liver. Intracytoplasmic lipid inclusions in the livers of both C57 and C3H starved mice have shown a deposition of reaction product for acid phosphatase, whereas only C57 starved mice have shown a deposition of reaction product for β-glucuronidase. Lipid droplets in the rat preputial gland were sites high for β-glucuronidase, as has previously been demonstrated by two dissimilar techniques. Particularly helpful in evaluating the results of electron microscopy has been the use of physiologic controls as, for example, by comparison of findings made with the "high" tissue β-glucuronidase strain, C57, versus those of the "low" tissue β-glucuronidase strain, C3H. Localization of electron-dense product in lysosomes was clear, but not all of this can be attributed unequivocally to enzyme-produced density. The limitations of the technique and the question of nonspecific uptake of diazotized mercuric aniline are discussed. The findings for both hydrolases support the concept of the dual localization of acid hydrolases in lysosomes and in endoplasmic reticulum.
IMMUNOHISTOCHEMICAL LOCALIZATION OF CATALASE IN MAMMALIAN TISSUES
Authors: MORIKAWA SHIGERU , HARADA TAKAYUKI
Published: April 2011

The distribution of catalase was investigated in bovine tissues using fluorescent antibody techniques. The immunochemical properties of liver catalase were also examined. Two distinct components of liver catalase in immune system were found. Both of them possessed common antigenicity with erythrocyte catalase. No cross-reactivity was observed by immunodiffusion between catalase and the other hemoproteins such as lactoperoxidase, cytochrome c and hemoglobins. Bovine liver, pancreas, kidney, spleen and peripheral blood were examined. Catalase was located mainly in the cytoplasm of hepatic cells, acinar cells of the pancreas, epithelia of proximal tubuli, splenic cells scattered in the red pulp and some leukocytes. It was not found in any nucleus. Intracorpuscular catalase could be revealed in the erythrocytes treated with surface-active agents but not in frozen sections.
STUDIES ON DEOXYRIBONUCLEIC ACID IN LEUKOCYTES AND RELATED CELLS OF MAMMALS VI. THE FEULGEN-DEOXYRIBONUCLEIC ACID CONTENT OF RABBIT LEUKOCYTES AFTER HYPOTONIC TREATMENT
Authors: GARCIA ALFREDO MARIANO
Published: April 2011

Peripheral blood of rabbit was subjected to the following hypotonic treatments in addition to controls: (1) 3:2 blood or plasma-distilled water; (2) 0.2 M sucrose; (3) 0.95% sodium citrate; (4) Ohnuki's medium ( Nature (London) 208: 916. 1965). The cells were smeared, fixed in methanol-acetic-formalin and stained by means of the Feulgen reaction. Cytophotometry was carried out in the Barr and Stroud integrating microdensitometer. The results show that the cells subjected to hypotonic media increase their chromatic load, the mean values being 7-12% higher than the controls. These results reinforce previous suggestions that any change in the physical state of deoxyribonucleoprotein molecules brings concomitant changes in the reactivity of their dye-binding sites and, moreover, that this increase is correlated with (1) diffusion of the deoxyribonucleoprotein complex and (2) increase of the mean nuclear size. Therefore, under the conditions used, the more diffuse the nuclear content, the higher the apparent deoxyribonucleoprotein value.
HISTOCHEMICAL AZO COUPLING REACTIONS OF THE PIGMENTS OF OBSTRUCTIVE ICTERUS AND OF HEMATOIDIN I. DIAZONIUM SALTS USED
Authors: LILLIE R. D. , PIZZOLATO P.
Published: April 2011

The azo coupling reaction readily demonstrates bile pigment and hematoidin in routine paraffin sections of human postmortem and surgical material fixed in formol, alcohol, chloroform + methanol, etc. A number of freshly diazotized amines as well as several commercial stable diazonium salts have been used successfully both in acid (1 N acetic acid) and slightly alkaline (pH 7.5-8) media. Alkaline coupling yields colors with some diazotates which have been perfectly stable for 14-18 months. Acid azo coupling almost completely eliminates background staining, but the staining results appear to be somewhat less stable. However, moderately good retention of stain for 6 months has been observed. Safranin O yields dark red to purple bile casts and granules and hematoidin globules and crystals, methylene violet (C.I. 50205) gives deep violet and p-nitroaniline (fast red GG) gives deep red colors of good stability. Fast black K, fast black B, fast blue B, fast garnet GBC and fast red B have given satisfactory red colors, but stable diazotates must be reasonably fresh with preferably under 1 year of shelf storage. Ethylanthranilate, anthranilic acid and even aniline have yielded deep red stains of bile casts and hematoidin, but the azo colors are increasingly unstable in the order given, the last fading completely in 18 hr. More consistent results are obtained with ethylanthranilate when a modified Claus diazotization is used. None of the diazonium salts tested discriminated between bile casts and hematoidin. The periodic acid-Schiff glycol reaction colors bile casts red but fails to color hematoidin globules. It is thought that this reaction demonstrates the presence or absence of glucuronic acid, although of course it is not specific for that substance. For the study of hematoidin several of the red azo coupling reactions were successfully combined with a preceding Prussian blue reaction for hemosiderin. Combination of the argentaffin reaction with the Prussian blue reaction does not prove practicable in either sequence. Sulfation enhances the basophilia of bile casts, but to a lesser degree than alkaline azo coupling with sulfanilic acid.
A FLUORESCENT METHOD FOR THE DETECTION AND LOCALIZATION OF ZINC IN HUMAN GRANULOCYTES
Authors: SMITH GARY L. , JENKINS ROBERT A. , GOUGH JAMES F.
Published: April 2011

A new cytochemical method is presented for the fluorescent demonstration of zinc in human granulocytes using 8-hydroxyquinoline. This compound is known to form fluorescent complexes with magnesium, calcium and zinc. Specificity for zinc is accomplished by adjusting the staining solution to pH 8.0 with Michaelis universal buffer. The technique is simple, rapid and much more specific for zinc than the dithizone stain.
PHYSICAL AND CHEMICAL PROPERTIES OF AMYLOID FIBERS II. ISOLATION OF A UNIQUE PROTEIN CONSTITUTING THE MAJOR COMPONENT FROM HUMAN SPLENIC AMYLOID FIBRIL CONCENTRATES
Authors: GLENNER G. G. , CUATRECASAS P. , ISERSKY C. , BLADEN H. A. , EANES E. D.
Published: April 2011

A major protein component (B protein) obtained from concentrated fibrils of each of two human amyloidotic spleens has been isolated by a series of fractionation methods using extraction in buffered 8 M urea, complete denaturation in buffered 6 M guanidine and column chromatography on Sepharose 4 B (Pharmacia). The major protein obtained by column chromatography was found to be also the major protein component on sodium dodecyl sulfate polyacrylamide disc electrophoresis. This protein was shown to have a molecular weight of 21,000-22,500. Immunologic identity was demonstrated for the B protein derived from the amyloid fibril concentrates of each spleen. No evidence of antibody cross-reaction to native or denatured components of normal human spleen or serum was discernible. It would appear, therefore, that the B protein derived from the two splenic amyloid fibril preparations, in addition to being the major protein component of the fibril, is not found in significant quantity in normal tissue or serum.
ELECTRON PROBE ANALYSIS OF CATIONIC SPECIES IN PYROANTIMONATE PRECIPITATES IN EPON-EMBEDDED TISSUE
Authors: LANE BERNARD P. , MARTIN EUGENE
Published: April 2011

Electron microscopy of Epon-embedded mouse vas deferens eipthelium treated with buffered potassium pyroantimonate-osmium tetroxide solution revealed precipitates in the lamina propria and along the apical plasma membrane. Electron microprobe elemental analysis of adjacent sections demonstrated that the deposits contained sodium and antimony. Other cations noted to precipitate pyroantimonate in vitro were not present in large amounts compared to controls, and were randomly distributed.
LEAD-ADENOSINE TRIPHOSPHATE COMPLEXES IN ADENOSINE TRIPHOSPHATASE HISTOCHEMISTRY
Authors: TICE LOIS W.
Published: April 2011

Chelation of lead by adenosine triphosphate (ATP) and its consequences for adenosine triphosphatase histochemistry were examined. The formation constant of lead-ATP chelates was found by two methods to be 4.6-4.7 x 104. The characteristics of enzyme inhibition by lead were consistent with the predicted effects of lead-ATP chelation. Inhibition was overcome by increasing ATP concentrations. With the adenosine triphosphatase from liver microsomes, which retained some activity in the presence of 4 mM Pb(NO3)2, substrate inhibition disappeared and increased MgCl2 was required for optimal activity. Increased solubility of lead phosphate was observed in the presence of increasing quantities of ATP in a manner predictable from lead-ATP chelation.
VALUE AND LIMITATIONS OF QUANTITATIVE CHEMICAL STUDIES IN INDIVIDUAL CELLS
Authors: GIACOBINI EZIO
Published: April 2011

Remarkable methodologic advances in recent years make it possible to study with high precision, sensitivity and accuracy the activities of most enzymes and the levels of most substrates and metabolites in individual cells. The limits and the value of quantitative chemical studies in individual nerve cells are critically evaluated and discussed.
ON THE NATURE OF THE HIGH IRON DIAMINE METHOD FOR SULFOMUCINS
Authors: GAD ADEL , SYLVÉN B.
Published: April 2011

Spectrophotometric and chromatographic studies on Spicer's high iron diamine reaction suggest that new red and blue oxidation products are formed, which become rapidly aggregated mainly to ferrous ions. Model experiments with pure substrates of known physical and chemical nature at pH 1.3-1.5 indicate that this reaction selectively demonstrates compounds carrying ester sulfate groups. Substrates with the highest charge density ( e.g., heparin and dextran sulfates) give dark purple or violet colors, whereas others with lower charge density give brownish reddish or purplish shades.
UPTAKE OF EXOGENOUS HORSERADISH PEROXIDASE BY COATED VESICLES IN MOUSE NEUROMUSCULAR JUNCTIONS
Authors: ZACKS SUMNER I. , SAITO ATUSHI
Published: April 2011

Following intramuscular injection of horseradish peroxidase into mouse skeletal muscle adjacent to areas of innervation, rapid uptake of the label was observed by coated but not synaptic vesicles. The tracer was also found in Schwann cells, external mesaxons and within peripheral myelin lamellae, but never in axoplasm of intramuscular nerves proximal to the injection site. The data suggest that the tracer is taken up by the coated vesicles and may be rapidly discharged into the synaptic clefts from which it is cleared by a combination of phagocytic activity and absorption via pinocytotic vesicles and intercellular clefts of adjacent capillaries or, more probably, enzyme activity is lost within the coated vesicles. The significance of these observations as related to neuromuscular function is discussed.
ULTRASTRUCTURAL LOCALIZATION OF INTRACELLULAR IMMUNE GLOBULINS IN PLASMA CELLS AND LYMPHOBLASTS BY ENZYME-LABELED ANTIBODIES
Authors: LEDUC ELIZABETH H. , SCOTT GEOFFREY B. , AVRAMEAS STRATIS
Published: April 2011

Techniques for localization of immune globulins by specific antibodies labeled with peroxidase or alkaline phosphatase were applied to spleens of rabbits hyperimmunized against various antigens. Best results were obtained with tissue blocks or thick sections after fixation in 1% formaldehyde (freshly prepared from paraformaldehyde) in cacodylate buffer with sucrose. The edges of blocks gave adequate penetration, comparable to thick sections. Ultrathin frozen sections which were reacted directly with labeled conjugate were less successful and glycol methacrylate sections failed to stain. Results were 3-fold: (1) immune globulins were demonstrated in ergastoplasmic cisternae, perinuclear space and Golgi apparatus of plasmocytes; (2) associated ribosomal staining varied somewhat with fixation and osmolarity of fixative; (3) lymphoblasts were shown to contain antibody. Generalized cytoplasmic staining was observed in the lymphoblasts, with no localization in the ergastoplasm and with no staining of control preparations. The significance of ribosomal and lymphoblast staining is discussed in relation to cell differentiation and the development of the immune response.
SOME CHARACTERISTICS OF THE COLD HYDROLYSIS TECHNIQUE FOR STAINING PLANT TISSUES BY THE FEULGEN REACTION
Authors: FOX DONALD P.
Published: April 2011

Hydrolysis of Vicia faba primary root meristems in 5 NHC1 at 20°C, followed by reaction with the Feulgen reagent, was investigated using microdensitometry. Some differences in response were observed with prophase nuclei fixed in different solutions, but in general this type of hydrolysis leads to a plateau for stainability. After plateau hydrolysis most dye binding is completed within 15 min. For 3:1 alcohol-acetic fixatives, fixation times between 1 day and 13 weeks do not affect plateau stainability but shorter (½ hr) and longer (21 weeks) times give slightly lower values for dye content. Alcohol-acetic fixatives for short periods of time give the least variable results. A comparison of 5 N HCl hydrolysis at 20°C and 30°C shows that at the higher temperature the plateau is brief but not different in dye binding. However, the peak values for 1 N HCl hydrolysis at 60°C are significantly lower by 20%.
OBSERVATIONS ON THE CHEMICAL REACTION BETWEEN BONE TISSUE AND CHLORAZOL FAST PINK BK
Authors: MÄKINEN KAUKO K.
Published: April 2011

Staining with Chlorazol Fast Pink BK (other trade names: Chlorantine Fast Red 2B, Solantine Pink 4BL, etc.) of human fetal parietal bones and skin collagen purified from the same fetuses was studied. In addition, the substantivity of the dye for achilles tendon collagen and for several other commercial substances was compared. The stained and homogenized bone tissue or collagen was treated with various chemicals, proteolytic enzymes and heat, and attempts were made to detect, using various chromatographic methods (gel filtration and thin layer chromatography), the nature of bonds between the stain and the bone components. The behavior of dye bone and dye collagen compounds was compared with that of the compounds between the dye and model substances, e.g., dextrans with varying degrees of cross-linking (Sephadex), cellulose, elastin, etc. The more tightly cross-linked dextrans adsorbed the dye more strongly than the less tightly cross-linked ones. The adsorption of the dye to purified collagen resembled in many respects its adsorption to bone. Chlorazol Fast Pink BK is bound to bone, collagen and the other substances tested chiefly through the formation of heat-labile and weak chemical bonds, preferably through hydrogen bonds or through van der Waals' attraction. The latter type of binding may be involved particularly in the reaction of the dye to cellulose. The adsorption of the dye to bone, collagen and other substances is favored by the elongated form of the reacting molecules. Under the conditions employed, there is also formation of salt type linkages between dye and collagen.
LOCALIZATION OF PEROXIDASE ACTIVITY IN MICROBODIES OF FETAL MOUSE LIVER
Authors: ESSNER EDWARD
Published: April 2011

The peroxidase activity of microbodies in fetal mouse liver was studied by light and electron microscopy. Two types of microbodies were present; a small population of bodies that lacked a nucleoid, predominant on the 16th day of gestation, and a larger population of nucleoid-bearing microbodies, predominant on the 19th day, in association with the rough endoplasmic reticulum from which they probably originate. Both types of bodies were visualized when incubated for peroxidase activity but were negative (19th day) for acid phosphatase activity. The findings suggest that the anucleoid- and nucleoid-bearing organelles together constitute the microbody population of the fetal liver.
ELECTRON MICROSCOPIC DEMONSTRATION OF ADENOSINE TRIPHOSPHATE PHOSPHOHYDROLASE ACTIVITY IN HERRING GULL SALT GLANDS
Authors: ABEL JOHN H.
Published: April 2011

Sites of adenosine triphosphate phosphohydrolase activity were demonstrated in herring gull salt glands with a cytochemical lead technique, utilizing fixed and unfixed tissue sections and a modified Wachstein and Meisel incubation media. The principal modifications in the procedure as recommended by Wachstein and Meisel included shortening the time of fixation and decreasing lead concentration while increasing the substrate concentration of the incubation media. Several different types of controls were run. In fixed tissues tested with an unmodified media, reaction product was deposited in capillary endothelial cells, tubular lumens and intercellular channels of central canal cells and nucleoli. Evidence is presented that none of these reactive sites contain a true adenosine triphosphate phosphohydrolase. With a modified procedure adenosine triphosphate-specific reaction product was deposited within the matrix of mitochondria, along the inner surface of the plasma membrane that lines the base of the principal salt-secreting cells and on the centriolar tubules. The possible validity of these reaction product localizations is discussed.
CHANGES IN ENZYME ACTIVITIES OF VARIOUS MUSCLE FIBER TYPES IN RAT INDUCED BY DIFFERENT EXERCISES
Authors: KOWALSKI K. , GORDON E. E. , MARTINEZ A. , ADAMEK J.
Published: April 2011

Mutability of enzyme activities (phosphorylase, succinic dehydrogenase, cytochrome oxidase) of red and white fibers was studied in rat quadriceps subjected to normal physiologic chronic exercises. A rise in phosphorylase activity was found in weight lifting and to a greater extent in running rats when muscle was taken as a whole, but both exercises resulted in equal increments for succinic dehydrogenase and cytochrome oxydase activities. Intraregional comparisons, however, revealed the greatest relative rise of succinic dehydrogenase activity in those fibers that were regarded as predominantly anaerobic in type. This effect was seen only with running and not with weight lifting. Statistically unproved but frequently observed after running in some of the preparations was a rise of phosphorylase activity in red fibers, although to a lesser degree than in white. Thus, in contrast to the dichotomy apparent in electrophysiologic events in nerve and its dependent muscle, metabolic demands may alter what is regarded normally as fixed fiber enzyme patterns. Whole muscle cannot be studied as a biochemical entity because of diverse regional responses to the same stimuli. Endurance exercise (running) and brief, forceful exercise (weight lifting) produced quantitatively different regional changes in succinic dehydrogenase and probably in phosphorylase activities.
PillCam COLON capsule endoscopy: recent advances and new insights
Authors: Adler Samuel N. , Metzger Yoav C.
Published: April 2011

Capsule endoscopy for the colon was introduced to allow an alternative screening method to the invasive classic colonoscopy. The results of initial studies have shown inferior detection rates of colonic polyps by the colon capsule. In this paper we aim to review and summarize the recent advances in wireless capsule endoscopy of the colon. Publications regarding the use of colon capsule with new technology, as well as personal experience, were reviewed. Since the introduction of the first generation of the colon capsule, many improvements have been made to create a better capsule endoscope. These include a wider angle of view, a faster adaptable frame rate and a new data recorder which is able to control the capsule activity during the transit through the bowel. Recent studies show these improvements had a direct effect on the colon capsule performance, leading to a better diagnostic yield. Recent advancements in the technology of the wireless colon capsule endoscope offer the option to screen patients for colonic polyps noninvasively. Colon capsule endoscopy may become relevant for assessment of extent of inflammatory bowel disease.
VOLUME AND DENSITY MEASUREMENTS IN SUBCELLULAR PORTIONS OF SINGLE NERVE CELL BODIES
Authors: LEHRER GERARD M. , KATZMAN ROBERT , WILSON CLARENCE E.
Published: April 2011

A simplified, improved method is described for the determination of volumes of microscopic tissue fragments. Such a method is imperative if measurements of concentrations of small, diffusible molecules are to be performed. A model system has been developed which establishes that the method is accurate within approximately 3%. The method has been applied to the determination of the dry mass per unit volume of fragments of cytoplasm and nuclei of puffer fish supramedullary neurons and of adjacent neuropil. The nuclei of these cells are large and watery and, as expected, their density is only 0.137 kg/liter. The cytoplasmic density is a uniform 0.224 kg/liter. That of adjacent neuropil is 0.195 kg/liter. A novel method is also described which greatly facilitates the handling and transportation of frozen-dried tissue fragments.
REDUCED PYRIDINE NUCLEOTIDE ABSORPTION IN INTRACELLULAR MITOCHONDRIA OF CULTURED LIVER CELLS
Authors: RITTER CARL , THORELL BO
Published: April 2011

In the 310-360 mµ range, reduced pyridine nucleotide in mitochondria which are inside human liver tissue culture cells shows an absorption maximum between 335 and 330 mµ. Thus, between one-third and two-thirds of the reduced pyridine nucleotide is protein-bound. Accompanying the changes at 330 and 335 mµ, there is a decrease in absorption at 265 mµ. The decrease in 265 mµ absorption which accompanies pyridine nucleotide reduction is more than would be expected based on the increase in absorption seen at 330-340 mµ. The extra decrease in 265 mµ absorption may have its origin in adenine nucleotide loss or decreased mitochondrial light scattering. A consideration of changes in 265 mµ absorption in cultured ascites tumor cells produced by glucose and by Dicumarol has led to the proposal that the main factor responsible for the large decrease in 265 mµ absorption is adenine nucleotide loss.
CLASSIFICATIONS OF ANTERIOR PITUITARY CELL TYPES WITH IMMUNOENZYME HISTOCHEMISTRY
Authors: NAKANE PAUL K.
Published: April 2011

Peroxidase-labeled antibody method was used to localize the six hormones of the anterior pituitary gland of male rats both at the light and electron microscopic levels. Growth hormone (GH), adenocorticotropic hormone (ACTH), prolactin and thyrotropic hormone (TSH) were found in separate cells. Follicle-stimulating hormone (FSH) and luteinizing hormone (LH) were frequently found in the same cell. TSH cells were scarce and were located at the periphery of the gland. The anterior-ventral portion of the gland contained few or no GH cells, ACTH cells, prolactin cells and TSH cells, but was filled with gonadotropic cells. In an area near the intermediate lobe, GH cells, ACTH cells and TSH cells were not found. GH cells and prolactin cells may be identified in electron micrographs without the aid of immunocytochemistry; however, ACTH cells and TSH cells may not be distinguished by their ultrastructural characteristics alone. Gonadotropic cells may be identified but their hormone content cannot be determined. The positive identification of these latter four cell types requires immunocytochemical methods.
ELECTRON MICROSCOPIC DEMONSTRATION OF ENERGY PRODUCTION AND COUPLED RESPIRATION OF IN SITU MITOCHONDRIA
Authors: KERPEL-FRONIUS S. , HAJÓS F.
Published: April 2011

Respiratory-linked accumulation of Sr2+ was applied to visualize mitochondrial energy production in unfixed slices of liver, kidney and heart muscle. The reaction was dependent on the function of the respiratory chain and energy coupling as was shown by the inhibitory effect of KCN and 2,4-dinitrophenol. The electron-dense granular or needle-like reaction product was localized in the inner compartment of the swollen mitochondria. Sr2+ translocation was also utilized to detect the coupled oxidation of different respiratory substrates. The overwhelming majority of mitochondria showed, more or less, activity when malate was oxidized. In the kidney and heart muscle a few mitochondria, however, did not show succinate-supported Sr2+ accumulation. This points to the possible differences in substrate utilization of mitochondria localized in the same cells. In the liver all mitochondria exhibited marked reaction with both substrates.
HISTOCHEMICAL DIFFERENTIATION OF TWO FORMS OF GLYCOGEN SYNTHETASE
Authors: SMITH ALLEN A.
Published: April 2011

Glycogen synthetase exists in two forms; one form is dependent on glucose 6-phosphate while the other is not. Variations in the levels of activities of the two forms of the enzyme were observed histochemically in active and inactive eccrine sweat glands from eight rhesus monkeys. The clear cells of active sweat glands showed less glucose 6-phosphate-independent glycogen synthetase activity than the clear cells of inactive sweat glands.
UPTAKE AND STORAGE OF CATECHOLAMINES IN MUCOSAL MAST CELLS OF THE RAT
Authors: ENERBÄCK LENNART , HÄGGENDAL JAN
Published: April 2011

The catecholamine-forming mast cells in rat intestinal mucosa have been studied by means of a histochemical fluorescence method and quantitative fluorometric analysis. Following the administration of l-DOPA, an accumulation of dopamine occurred in these cells which could be prevented by pretreating the animals with a DOPA decarboxylase inhibitor. Evidence is presented which indicates a decarboxylation of DOPA by an intracellularly located enzyme. d-DOPA was less effective in producing catecholamine fluorescence in the mucosal mast cells, thus indicating a stereo-specific uptake or decarboxylation of DOPA. No evidence for an enzymatic degradation of dopamine in the mast cells was obtained, as judged by the effect of monoamine oxidase inhibitors or a catechol-O-methyl transferase inhibitor on the fluorescence intensity. The weak, naturally occurring fluorescence in the mucosal mast cells may be due to dopamine but the properties of the fluorescence are also compatible with a low concentration of 5-hydroxytryptamine.
DISCONTINUOUS ELECTROPHORESIS OF PEPSIN AND PEPSINOGEN IN THIN SHEETS OF POLYACRYLAMIDE GEL
Authors: CUNNINGHAM LEW , RASCH ELLEN M. , LEWIS ANN L. , HEITSCH RICHARD
Published: April 2011

Electrophoretic separation of discrete protein bands from solutions of crystalline pepsin or pepsinogen was accomplished in a system incorporating pH discontinuity and using standard, glass microslides to support thin (0.25-mm) sheets of 15% polyacrylamide gel. Adherence of gel films to a glass support during fixation and staining with crystal violet not only prevented distortion of gels as a result of swelling, but also provided preparations that could be readily scanned with a microspectrophotometer. A number of discrete peptide bands were identified from different samples of purified pepsin. Among the several heterogeneous components found in commercially available samples of crystalline pepsinogen, there was a characteristic impurity, the mobility of which was directly comparable to that of pepsin run under the same conditions of electrophoresis. Specific details of methodology are presented.
ULTRASTRUCTURAL LOCALIZATION OF CHORIONIC GONADOTROPIN IN HUMAN TERM PLACENTA
Authors: DRESKIN R. B. , SPICER S. S. , GREENE W. B.
Published: April 2011

Human chorionic gonadotropin (HCG) was localized in human placenta at the ultrastructural level by an immunocytochemical method which linked the antigen HCG to a peroxidase label through the antigen-antibody reactions of an immunoglobulin-peroxidase bridge. Material on the maternal surface of the apical plasma membrane and in the cisternae of rough endoplasmic reticulum of the syncytiotrophoblast displayed strong immunostaining for HCG and the outer surface of the basal plasma membrane appeared equivocally immunoreactive. Lamellae and vesicles of the Golgi complex of the syncytium lacked electron-opaque reaction product indicative of immunostaining for HCG. Deposits of peroxidase reaction product in the large variety of cytoplasmic dense granule varied from negligible to abundant in immunostained and in control specimens alike and, accordingly, the large granules appeared to contain intrinsic peroxidase but not HCG. The question of the immunoreactivity of the small variety of cytoplasmic dense granule was not conclusively settled, but the occurrence of reactive and unreactive small granules in both immunostained and control specimens weighed against the presence of HCG in these rather infrequent granules. The cytotrophoblast lacked evidence of immunoreactivity.
CYTOTOXIC EFFECTS OF PUROMYCIN ON THE GOLGI APPARATUS OF PANCREATIC ACINAR CELLS, HEPATOCYTES AND AMELOBLASTS
Authors: WEINSTOCK ALFRED
Published: April 2011

A single injection of puromycin was given to rats in a dose sufficient to produce almost total inhibition of protein synthesis in pancreas and liver. By 10 min after injection, Golgi saccules in pancreatic acinar cells, hepatocytes and ameloblasts appeared grossly and irregularly distended, and almost devoid of content. In pancreas the condensing vacuoles near the inner face of the Golgi were often altered, and those normally present in ameloblasts were lacking. Between 2 and 3 hr after injection, protein synthesis had started anew. At this time, granules without limiting membranes appeared within cisternae of the rough endoplasmic reticulum in acinar cells and ameloblasts. These intracisternal granules are believed to consist of newly synthesized secretory protein which could not be transported through the disrupted Golgi apparatus to be packaged into secretory granules. Indeed, by 3 hr postinjection the secretory granules which normally abound in the apical processes of ameloblasts were sparce or absent. Thus, while biochemical evidence indicates that puromycin blocks protein synthesis on the ribosomes, the use of this antibiotic in vivo results in alterations in the Golgi apparatus and interruption of the packaging of protein into secretory granules.
MICROBODY FORMATION IN REGENERATING RAT LIVER
Authors: RIGATUSO JOSEPH L. , LEGG PETER G. , WOOD RICHARD L.
Published: April 2011

The formation of new microbodies induced by partial hepatectomy was studied in the liver of male rats using conventional microscopy and the peroxidase reaction for demonstrating catalase. In the early phases of the regenerative period, small smooth walled vesicles were identified adjacent to mature microbodies and these contained material similar both in appearance and in endogenous peroxidase activity to the neighboring microbody. Membrane continuity between the two organelles was identified clearly. It is suggested that the formation of new microbodies from pre-existing microbodies may be an important general method of microbody proliferation.
STUDIES OF ACID PHOSPHATASE ISOENZYMES IN HUMAN LEUKOCYTES
Authors: LI C. Y. , YAM L. T. , LAM K. W.
Published: April 2011

Seven acid phosphatase isoenzymes, namely 0, 1, 2, 3, 3b, 4 and 5, in human leukocytes have been separated both by acrylamide gel electrophoresis and by ion exchange chromatography. The study of these isoenzymes in preparations of various cell types indicates that at least some of the isoenzymes are cell type-specific, i.e., isoenzyme o in Gaucher cells, 1, 2 and 4 in neutrophils, 1 and 4 in monocytes, 3 in lymphocytes and platelets, 3b in primitive blasts and 5 in reticulum cells of leukemic reticuloendotheliosis. Isoenzyme 5 is different from others in its resistance to l(+)-tartaric acid treatment. The persistent presence of abnormally high isoenzyme 5 in the reticulum cells of leukemic reticuloendotheliosis may be important for the differential diagnosis of this disease. Possible relationships between these isoenzymes and subcellular organelles are discussed.
AN ULTRASTRUCTURAL HISTOCHEMICAL STUDY OF BROWN ADIPOSE TISSUE FROM NEONATAL RATS
Authors: SUTER ELSI R. , STÄUBLI WILLY
Published: April 2011

Mitochondrial matrix inclusions in brown fat from neonatal rats appear to consist of lipid. They are absent after lipid extraction of isolated mitochondria. The inclusions are preferentially bleached when reduced osmium is oxidized with diluted H2O2 or periodic acid. In glutaraldehyde-fixed tissue the inclusions appear in negative image. They are invisible after "preferential staining for ribonucleic acid." Inclusions are unaffected when protein is extracted by digestion with Pronase. No cytochrome oxidase activity appears to be associated with the inclusions. The activity of the inner membrane seems to be rather low in mitochondria from neonatal rats. In brown fat from 3-week-old rats, which has the largest mitochondria with maximally crowded cristae, the activity appears greatly increased, perhaps beyond the increase in inner membrane. The postnatal changes in mitochondrial structure, among which the disappearance of inclusions is the most striking, are probably mediated by cyclic adenosine monophosphate. The nucleotide has no direct effect on isolated mitochondria, and its mechanism of action remains to be elucidated.
IMMUNOFLUORESCENT LOCALIZATION OF VITAMIN D-DEPENDENT CALCIUM-BINDING PROTEIN
Authors: TAYLOR ALAN N. , WASSERMAN ROBERT H.
Published: April 2011

The vitamin D-induced calcium-binding protein (CaBP) present in chick intestinal mucosa was purified and used for the preparation of an antiserum in rabbits. After adsorption the antiserum was judged to be specific for CaBP by immunoelectrophoresis in three different systems. In addition the antiserum was capable of inhibiting the calcium-binding activity of CaBP as measured by equilibrium dialysis. Using the antiserum in micro-Ouchterlony double diffusion tests the previously reported tissue distribution of CaBP in vitamin D-replete chicks was confirmed, i.e., all segments of small intestine and kidney. In addition, the protein was detectable in homogenates of colon from vitamin D-treated chicks, but not in blood plasma, liver, pancreas or bone. The sensitivity of the double diffusion reaction indicated that if CaBP were present in bone it would be at a concentration less than 10 µg/g bone cells (dry weight). Fluoresceinisothiocyanate-conjugated sheep-anti-rabbit γ-globulins were used in the indirect fluorescent antibody localization of CaBP in 4-6-µ cryostat-prepared sections of chick intestinal tissue. Specific fluorescence, indicative of the presence of CaBP, was noted in the periodic acid-Schiff-positive goblet cells and in association with the surface coat-microvillar region of all intestinal epithelial cells in sections from normal and vitamin D-treated rachitic chicks. Immunologically blocked controls and sections from rachitic chicks demonstrated nonspecific fluorescence only.
LOCATION OF ANTIBODY TO HORSERADISH PEROXIDASE IN POPLITEAL LYMPH NODES OF RABBITS DURING THE PRIMARY AND EARLY SECONDARY RESPONSE
Authors: STRAUS WERNER
Published: April 2011

After a primary injection of horseradish peroxidase into the footpads of rabbits, the specific antibody reaction in popliteal lymph nodes was first seen in plasma cells of medullary cords and, later, in lymphoblasts of germinal centers in the cortex. During the late primary or early secondary response, the reaction also became positive in reticular cells and lymphocytes. Specific antibodies were also present in the intercellular spaces between lymphocytes (reticulum) in the lymphoid follicles and in globules distributed throughout the lymph node. Two weeks following a primary injection of the antigen, many plasma cells containing the specific antibody were located in proximity of macrophages and reticular cells. The phagolysosomes of many macrophages and reticular cells became antibody-positive at about the same time. Double staining procedures were developed by which the antigen and antibody and the antigen or antibody and acid phosphatase (lysosomes) could be visualized in the same tissue section.
HISTOCHEMISTRY OF HISTAMINE: MICROSPECTROFLUOROMETRIC CHARACTERIZATION OF THE FLUOROPHORES INDUCED BY o-PHTHALDIALDEHYDE
Authors: HÅKANSON R. , JUHLIN L. , OWMAN C. , SPORRONG B.
Published: April 2011

o-Phthaldialdehyde (OPT) has been used as a reagent for the histochemical demonstration of histamine by fluorescence microscopy. OPT induces a blue fluorescence in gastric epithelial cells of the rat and, under the same conditions, a yellow fluorescence in mast cells. Microspectrofluorometric analysis of the histamine-OPT fluorophore in model systems (silica gel thin layer or dried albumin droplets) revealed a concentration-dependent shift of the emission maximum from blue (at low histamine concentrations) to yellow (at high concentrations). The characteristics of the fluorophores, which were very ultraviolet-labile, were not overtly affected by pH, but the red shift was more pronounced upon hydration. The blue fluorescence was maximally excited by the 365 mµ line of the mercury lamp; the excitation maximum for the yellow fluorescence was approximately 405 mµ. This strongly suggests that at least two different fluorophores are formed as a consequence of the condensation between histamine and OPT. This was confirmed by thin layer chromatography of the OPT-histamine fluorophore and by the finding that ultraviolet exposure converts the yellow fluorophore into a blue one. It is suggested that the blue and yellow OPT-induced fluorescence in cells reflect markedly different concentrations of histamine. The present observations may offer a new possibility for the histochemical quantitation of cellular histamine based on the concentration-dependent shift of the emission and/or excitation maxima.
ELECTROPHORETIC SEPARATION OF ESTERASES OF PULMONARY ALVEOLAR CELLS
Authors: SCHIFF ROBERT , BRUNSTETTER MARY ANN , HUNTER ROBERT L. , CROSS CARROLL E.
Published: April 2011

Starch gel electrophoresis and vertical flat bed electrophoresis in polyacrylamide gels were used to separate butyrylesterases of pulmonary alveolar macrophages (PAMs) from RF/Al(+) and RF/Al(–) mice. PAM esterases from (+) mice showed five bands in starch and five or six in acrylamide whereas four and three or four bands, respectively, were found in extracts from (–) animals. The additional band or bands present only in positive samples corresponded to the Es-2 prealbumin serum esterase found only in the RF/Al (+) animals. This isozyme was more sensitive to diisopropylfluorophosphate than any other PAM esterase. The serum counterpart was sensitive to the same degree. None of the macrophage esterases were inhibited by eserine sulfate. Spectrophotometric analysis of serum esterase activity indicated a statistically significant increase in female mice from both sublines as compared to their respective males, but did not show a difference between the two sublines. The usefulness of the esterase marker in studies of PAM origin and pathologic conditions is discussed.