bluuebunny/crossref_metadata_2025_split huggingface dataset:
LOCALIZATION AND IDENTIFICATION OF TESTICULAR ESTERASES IN THE RAT
Authors: NIEMI MIKKO , HÄRKÖNEN MATTI , KOKKO AULIKKI 
Published: April 2011

Both the Leydig and Sertoli cells of the rat testis have been shown to be rich in non-specific esterase, but lacking in cholinesterase and "true" lipase. No esterolytic activity has been observed in other elements of testicular tissue. The enzyme activity of the Sertoli cells is mainly localized to the long cytoplasmic processes of these cells, where the sperm heads are embedded. The distribution of esterase activity inside the seminiferous tubules undergoes cyclic variations corresponding to those seen in the morphology of the Sertoli cells. An essentially similar distribution of esterase activity was noticed when α-naphthyl acetate, naphthol-AS acetate, o-indoxyl acetate or Tweens 20 to 60 were used as substrates. Indoxyl acetate was, however, hydrolysed even in an acid medium and this activity was, furthermore, resistant to an organophosphorus inhibitor (E600). It is therefore concluded that the testicular Leydig and Sertoli cells have both A-type and B-type esterase activity, of which the former behaves much like an intracellular peptidase. Demonstration of the activity of electrophoretically separated enzyme proteins have further shown that the histochemically demonstrated non-specific esterase activity in the testis is composed of about eight different enzyme proteins, whose behavior towards inhibitors corresponds to that of tissue sections.
IMPROVED TECHNIQUES FOR THE ENZYMATIC EXTRACTION OF NUCLEIC ACIDS FROM TISSUE SECTIONS
Authors: AMANO MINORU 
Published: April 2011

Since the removal of only one of the two nucleic acids, ribonucleic and deoxyribonucleic, is often necessary in biological work, the optimum conditions for the specific removal of either nucleic acid by the corresponding enzyme were investigated. The specific removal of nucleic acids by enzyme treatment was tested by toluidine blue staining and by quantitative study of the radioautographic reactions produced by the labelled deoxyribonucleic acid and ribonucleic acid appearing in tissues of a cytidine-H3 injected mouse. For the removal of ribonucleic acid by ribonuclease, the tissues must be fixed in freshly prepared Carnoy's solution for 24 hours, as a shorter fixation permits the loss of ribonucleic acid from control tissue sections. Ribonuclease extraction (1 mg crystallized salt-free Worthington ribonuclease per 1 ml of distilled water) is carried out for 4 hours at 40°C. For the removal of deoxyribonucleic acid by deoxyribonuclease, it was also necessary to use 24 hour fixation in freshly prepared Carnoy. A high concentration of magnesium was found to be necessary in the incubation solution (0.05 mg crystallized Worthington deoxyribonuclease per 1 ml of 0.2 M Mg-containing Gomori's Tris maleic acid buffer solution). The incubation was carried out for 24 hours at 37°C. The toluidine blue stained basophilic material and the radioautographic reactions in the tissues of the cytidine-H3 injected mouse were lost following a double treatment with ribonuclease and deoxyribonuclease or by hot trichloracetic acid extraction. Therefore all the reactions appearing over the histological tissue sections are due to the tritium incorporated into deoxyribonucleic acid and ribonucleic acid. The basophilic material from cytoplasm and nucleolus was removed specifically and completely by ribonuclease treatment. The deoxyribonuclease treatment removed nuclear basophilia exclusively. However, controls of ribonuclease or deoxyribonuclease treatment showed no loss of basophilia, except on occasion in the pancreas. The radioautographic reactions found in the mouse tissues 24 hours after cytidine-H3 injection were exclusively localized over the nuclear basophilia of some nuclei after ribonuclease treatment, but were largely over the cytoplasmic basophilia after deoxyribonuclease treatment. Quantitative radioautographic study showed that either enzyme treatment removed its specific substrate completely and exclusively without loss in control preparations.
IMPROVEMENTS IN THE COATING TECHNIQUE OF RADIOAUTOGRAPHY
Authors: KOPRIWA BEATRIX MARKUS , LEBLOND C. P. 
Published: April 2011

The 1958 coating technique of Messier and Leblond for radioautography—the most recent modification of the liquid emulsion method—was investigated in the hope of eliminating artefacts and discovering the optimum conditions for reliable quantitative radioautography. From a review of the properties of the various bulk emulsions available, it is concluded that NTB2 bulk emulsion is the most suitable. The proper handling of the emulsion (transportation, storage and testing operations) is described, as well as the details of an improved coating techniqe. Comparison is made of this method with the other most widely used, the strippingfilm technique. The main features of the improved coating technique are as follows: 1. Various fixatives may be used for radioautography. Zenker and presumably other mercury-containing fixatives are unreliable, but can be used after complete removal of mercury salts. 2. Dipping the sections into celloidin prevents the infiltration of the tissue by emulsion, but is otherwise unnecessary and may be omitted for routine work. 3. Emulsion coating is done by dipping sections in a bubble-free emulsion at 40°C. 4. Drying of the coated preparations is best accomplished by keeping them vertical over moistened tissue paper at 28°C and 80% relative humidity. 5. Exposure is done in a dry atmosphere at 4°C. 6. Processing is carried out at 17-18°C, as even a slight rise in temperature increases the background fog and decreases the efficiency of the emulsion. The optimal duration of development is 2 minutes for D-72 (Dektol), 6 minutes for D-l70 (Dolmi) and 3 minutes for D-178. For fixation, best results are obtained using acid fixer with hardener (or 24% hypo) for 3 minutes. 7. Mounting is done by successive immersions in an alcohol-cedar oil mixture, a xylene-balsam mixture, and balsam. Malinol may replace balsam. The improved coating technique arising from these observations was tested for its quantitative response, measured by grain counts. The suitability of the technique for quantitative radioautography is demonstrated by the fact that the reaction intensity is proportional to exposure up to at least 80 days with hematoxylin-cosin stained sections, and 1 year with unstained sections. The background increases slowly with time, but the increase is more rapid on slides with C14-labelled sections than on those labelled with H3. In conclusion, the improved coating technique using the NTB2 emulsion is now quite satisfactory. The next major improvement would be for the industry to provide emulsions with a standardized sensitivity and a constant, low level of fog. Nevertheless, in its present state, it is a convenient method permitting the preparation of large numbers of high quality radioautographs and is well suited to the grain counting required for quantitative radioautography.
HISTOCHEMISTRY AND ELECTRON MICROSCOPY OF MITOCHONDRIAL ADENOSINETRIPHOSPHATASE
Authors: LAZARUS SYDNEY S. , BARDEN HERBERT 
Published: April 2011

A study of the requirements for histochemical demonstration of mitochondrial adenosinetriphosphatase demonstrated that fixation of cryostat sections for 15 minutes in cold 0.05 M manganese chloride in 3% formol saturated with CaCO3 (pH 6.9) gave best results. By this means the enzyme could be demonstrated with both the calcium and lead incubation media. The adenosinetriphosphatase activity was demonstrated ultramicroscopically and was found to be compartmentalized in the intereristal matrix. The histochemically demonstrated enzyme showed responses to inhibitors and activators characteristic of adenosinetriphosphatase of damaged mitochondria obtained by ultracentrifugation.
A METHOD FOR THE HISTOCHEMICAL DEMONSTRATION OF DISACCHARIDASE ACTIVITIES: APPLICATION TO INVERTASE AND TREHALASE IN SOME ANIMAL TISSUES
Authors: DAHLQVIST ARNE , BRUN ARNE 
Published: April 2011

1) A histochemical method for the demonstration and localization of disaccharidase activity using various disaccharides as the substrate is described. Frozen sections are incubated in a solution containing substrate plus a coupled oxidation-reduction system of glucose oxidase, phenazine methosulfate and a tetrazolium salt. Where disccharidase activity is present, the substrate is hydrolyzed with the liberation of glucose, and a deep blue insoluble formazan is formed. 2) This method has been used for the demonstration of invertase and trehalase in rat tissue sections. The small intestine, colon, stomach and kidney were investigated. 3) All the epithelial cells of the small intestinal mucosa, in the villi, crypts and glands of Brunner, contain invertase and trehalase. These enzymes appear to be localized to small rounded granules which are scattered throughout the cytoplasm of the cells. The activity is especially high in the perinuclear part of the epithelial cells, but no concentration of the granules is to be seen in "brush border" region. 4) The stomach and the kidney also contain weak invertase activity which is present in granules localized to small distinct cell groups. No trehalase activity could be demonstrated in these organs. The colon contains essentially no invertase or trehalase.
ON THE BINDING OF CONGO RED BY AMYLOID
Authors: PUCHTLER HOLDE , SWEAT FAYE , LEVINE M. 
Published: April 2011

Congo red in alkaline 80% ethyl alcohol solution saturated with NaCl stains amyloid selectively. The distribution of the dye is very similar to that observed in serial sections stained with Highman's Congo red method. In sections stained with Congo red in aqueous or alcoholic solutions without differentiation all tissue structures are intensely colored, but only amyloid is dichroic. The rapid removal of dye by alkaline solutions from structures other than amyloid suggests ionic or salt-type bonds between Congo red and these structures. In contrast, the binding of Congo red by amyloid from alkaline alcoholic solutions and the increase in intensity of staining upon addition of NaCl indicate a non-ionic type linkage between amyloid and dye. The effects of deamination, acetylation and various oxidation procedures on the binding of Congo red indicate interaction of the dye with hydroxyl groups of amyloid. According to data on the direct dyeing of cellulose in textile dyeing, Congo red is adsorbed by hydrogen bonding between hydroxyl groups of the polysaccharide chains and the amino groups of the dye. It seems probable that a similar mechanism is involved in the binding of Congo red by amyloid. It is suggested that the selective staining of amyloid with Congo red is due to steric configurations of amyloid or one of its components which favor non-ionic binding of the eye.
STUDIES IN THE HISTOCHEMISTRY OF PLASMALOGENS: I. THE EFFECT OF FORMALIN AND ACROLEIN FIXATION ON THE PLASMALOGENS OF ADRENAL AND BRAIN
Authors: NORTON WILLIAM T. , GELFAND MARTIN , BROTZ MIRIAM 
Published: April 2011

Formalin containing fixatives were found to cause a rapid drop in extractable plasmalogens of steer adrenal cortex and medulla and rat brain to 50-60% of the initial value in 4-6 hours. This rapid destructive reaction was followed by a slow decrease so that for several days the plasmalogen levels remained relatively constant. An acrolein containing fixative, although not evaluated with adrenal, showed similar behavior with rat brain. Control experiments show the fixative aldehyde is solely responsible for this plasmalogen loss. This decrease is not paralleled by a similar decrease in extractable total lipid or total phospholipid. It is believed that this decrease represents an actual destruction of the enol-ether bond by chemical reaction with the fixative aldehyde. Although the nature of this reaction is unknown, model experiments show cephalin plasmalogen to be much more labile than choline plasmalogens to formaldehyde, although all the cephalin plasmalogens are not destroyed during the fast reaction in tissue. These experiments also show that steer adrenal cortex and medulla have the same plasmalogen content and that the plasmalogen in either part of the gland responds the same to fixation, even though plasmal staining would indicate that the medulla is richer in plasmalogen. After a period of fixation sufficient to render the plasmal reaction negative in both adrenal cortex and medulla and in rat brain there is still at least 35-50% of the initial plasmalogen present. The intensity of the plasmal stain appears to parallel the concentration of the fixative labile plasmalogen. In these particular tissues there was no suggestion of an increase in free aldehyde over plasmalogen until the tenth day of fixation.
THE CYTOCHEMICAL DEMONSTRATION OF SUCCINIC DEHYDROGENASE IN MOUSE LEUKOCYTES
Authors: MORRISON JOHN H. , KRONHEIM SUZANNE 
Published: April 2011

The specificity of Nitro BT as an electron acceptor for the intracellular localizations of succinic dehydrogenase in mammalian blood cells was investigated. Imprints of mouse spleen were fixed either by immersion in acetone or formol calcium under varying conditions of time and temperature or by freezing in cold isopentane. The incubation mixture used for the detection of enzyme activity contained succinate, Nitro BT, amytal, and phosphate buffer (pH 7.4). By comparison of the intensity and site of formazan production in imprints prepared under varying conditions of fixation, it was found that specific intracellular localization of succinic dehydrogenase occurred only in cells fixed in formol calcium prior to, or after incubation with Nitro BT. Using this fixation method, succinic dehydrogenase was detected in the hemocytoblast, lymphocyte, myelocyte, and megakaryocyte. In all these cell types the enzyme was confined exclusively to the mitochondria. Only slight activity was observed in occasional metamyelocytes and none in any of the mature segmented forms. Imprint cells fixed in formol calcium failed to exhibit endogenous enzyme activity when succinate was omitted from the incubation medium (control series). It was also shown that reduction of Nitro BT was inhibited completely in the presence of 6 mM malonic acid (inhibition series) and that this inhibition was reversible in the presence of excess succinate (reversal series). The results of this study are discussed in terms of their possible significance in the differentiation process leading to formation of the mature granulocyte. On the basis of our findings and other information in the literature concerning modifications in fine structure of the mitochondria during blood cell maturation, it was suggested that cytogenesis of the granulocyte is associated with a progressive loss in oxidative enzyme activity and that oxidation reactions in the mature granulocyte may be mediated, for the most part, by glycolysis.
AN IMPROVED APPARATUS FOR QUANTITATIVE CYTOCHEMICAL MODEL STUDIES AND ITS USE IN AN EXPERIMENTAL TEST OF THE TWO-WAVELENGTH METHOD
Authors: VAN DUIJN P. , DEN TONKELAAR E. M. , HARDONK M. J. 
Published: April 2011

The construction of an improved apparatus for quantitative studies on cytochemically-stained membranes is described. The membrane-holder of this apparatus was constructed in such a way that the area of the film whose extinction is measured coincides exactly with the area punched out for weight determination and chemical analyses. In addition, provisions were made to assure an uniform response of the photomultiplier to light rays traversing different parts of the field. Using this system, an experimental test of the so called two-wavelength method was carried out. The spectra of Feulgen-stained deoxyribonucleic acid cellulose films or periodic acid Schiff-stained cellulose films were measured. The stained membranes were punched out, after which the resulting discs were cut, folded, and brought back to their original position in the holder between two unstained cellulose membranes. In this way the number of chromophores in the measured area was kept constant although their macroscopic distribution had changed considerably. From the spectra of the folded stained discs the extinction of the same material in unfolded form could be calculated by applying the two-wavelength method of Patau and Ornstein. The two-wavelength method was found to correct well, but some residual error remained even in this macrosystem, due to the relatively strong effect of small measuring errors on the end results.
DISTRIBUTION OF PROTEIN-BOUND SULFHYDRYL GROUPS IN YEAST CELLS
Authors: POMERANZ Y. 
Published: April 2011

Young and old cultures of 12 yeasts have been stained by the Barrnett-Seligman histochemical procedure for localization of sulfhydryl groups. Thiol groups have been found distributed in the cytoplasm, but not in the cell wall. The intense staining of sulfhydryl groups by the specific stain was clearly showns in the slender thread of protoplasm between bud and mother cell. The intensity of staining in the proliferating cell was higher than in the resting cell; variations in thiol content between various yeast species were recorded. No decrease in thiol content of yeast spores, as compared with vegetative cells, has been found in Schizosaccharomyces octosporus, whereas the staining of yeast spores in Saccharomyces and Hansenula was found to be negligible. Implications of these findings are discussed.
TWO-EMULSION RADIOAUTOGRAPHY
Authors: BASERGA RENATO , NEMEROFF KEN 
Published: April 2011

Two-emulsion radioautography can distinguish between beta particles emitted from C14 atoms and those originating in H3 atoms. It uses two layers of sensitized emulsion separated by an inert layer of celloidin: the C14 beta particles, because of their longer range, are recorded in the second emulsion, but the beta particles from tritium are arrested by the first emulsion. Experiments have shown that the combination of two NTB emulsions separated by a celloidin layer is the most satisfactory. The total thickness of the radioautograph is 19 µ. The first emulsion plus the celloidin layer measure together 12 µ in thickness and decrease by 38% the flux of C14 beta particles reaching the second emulsion. Staining of the specimen with Mayer's hematoxylin and eosin and celloidin coating follow the exposure and processing of the first emulsion. The second emulsion is applied after the celloidin coating and processed after a suitable second exposure. By using a tritiated precursor of DNA and a C14-labeled precursor of RNA or proteins, or viceversa, two-emulsion radioautography can be applied to investigate two distinct metabolic processes occurring at the same time in the same cell.
HISTOCHEMICALLY SELECTIVE ACIDOPHILIA OF BASIC NUCLEOPROTEINS IN CHROMATIN AND NUCLEOLI AT ALKALINE pH
Authors: SPICER S. S. 
Published: April 2011

Chromatin and nucleoli of tissues fixed with formalin-free, mercurial or alcoholic solutions, have the unique property of staining metachromatically with the acid dye Biebrich scarlet in the restricted pH range 9.0-10.0. The nuclear structures lack affinity for fast green FCF under these conditions so that a mixture of Biebrich scarlet and fast green stains chromatin and nucleoli a characteristic orange-red at pH 9.5. A Feulgen-Biebrich scarlet sequence distinguishes magenta DNA in chromatin and orange-red basic protein in nucleoli. Although nuclei in tissues fixed with the buffered mercurial show strong affinity for the basic dye alcian blue, nuclei in formalin-fixed tissue do not. In formalin-fixed, trichloroacetic acid (TCA)-extracted tissues, chromatin stains with Biebrich scarlet or fast green but nucleoli do not. The acidophilia induced by TCA may in part depend on the fact that deoxyribonucleic acid (DNA) is removed by this treatment. However such tissues lose the acidophilia of the chromatin when treated again with formaldehyde but regain the acidophilia after re-extraction with TCA, indicating removal of formaldehyde condensates from amines by TCA. The affinity of chromatin and nucleoli for Biebrich scarlet at pH 9.5 is highly susceptible to destruction by relatively brief nitrosation, acetylation or exposure to formaldehyde. Extraction with a sulfuric acid-mercurial reagent also eliminates this staining; but other solvents used for isolation of histone do not, except that 1 N HCl has such an effect when employed on sections of tissue fixed with HgCl2. After mild treatment with TCA, exposure to pH 9.5 buffer extracts DNA whereas buffer at pH 6.0 has no such effect.
THE ACTION OF FORMALDEHYDE SOLUTIONS ON HUMAN BRAIN LIPIDS
Authors: HESLINGA F. J. M. , DEIERKAUF F. A. 
Published: April 2011

The composition of the lipids extracted from fresh normal human brain and from brains preserved up to 24 years in unbuffered formaldehyde solutions was analysed quantitatively using column and paper chromatography, followed by various determinations. The amounts of cholesterol, cerebrosides, sulphatides, phosphoinositides and sphingomyelin remain unaffected. Lecithin, phosphatidylethanolamine and phosphatidylserine are broken down to the corresponding lyso compounds, fatty acids, phosphatidic acid and probably lysophosphatidic acid. The lyso compounds are further broken down by the liberation of the second fatty acid. By this process the fatty acid content is markedly increased. The total lipid content decreases slightly by the formation of water soluble phosphoryl compounds. Consequently the phosphorus content of the total lipid extract suffers marked diminution. Histochemical reactions on lipids in formaldehyde stored tissues should preferably be performed in combination with an analysis of the lipid extract.
THE MECHANISM OF TISSUE STAINING BY FERRIC HYDROXYQUINOLINE METHODS FOR β-GLUCURONIDASE
Authors: JANIGAN D. T. , PEARSE A. G. E. 
Published: April 2011

1. The investigations reported in this paper show that the staining product of the Fishman-Baker and other ferric hydroxyquinoline methods for the demonstration of β-glucuronidase is entirely uninfluenced by the presence or absence of the enzyme. 2. Staining is due to the uptake of ferric iron from the complex equilibrium mixture of the incubating medium. 3. The uptake of iron is influenced by a number of factors but is essentially dependent on the nature of the tissue. 4. Changes in the localization of iron staining which occur in tissues in physiological or pathological states are likely to be due to significant though ill-understood differences in affinity for iron shown by intracellular constituents.
THE DISTRIBUTION OF CHOLINESTERASES IN THE SENSORY GANGLIA OF MAN AND OF SOME MAMMALS
Authors: CAUNA N. , NAIK N. T. 
Published: April 2011

The distribution of cholinesterases in the cranial and spinal sensory ganglia was investigated in the cat, guinea-pig, rat, mole and man using a modified Koelle's histochemical technique. It was found that the cytoplasm of the sensory nerve cells contained acetylcholinesterase in all species, but the concentration of the enzyme varied from cell to cell. Nerve fibers gave a negative acetylcholinesterase reaction in the cat and guinea-pig, and a weak to moderate positive reaction in the rat, mole and man. Non-specific cholinesterase was only found in the sensory ganglia of the cat and guinea-pig. In human sensory ganglia, it was found during the fetal life but not after birth. The sensory ganglia of the rat were found to be nearer to those of man than the ganglia of the cat or guinea-pig in respect of cholinesterase distribution. It is suggested that the variations in acetyl-cholinesterase activity among different nerve cells may signify the existence of several cell types in the sensory ganglia. The role of intracytoplasmic cholinesterase in the sensory nerve cells is discussed.