Showing papers in "Journal of Histochemistry and Cytochemistry in 1953"
TL;DR: The general histochemical procedure of Linderstrøm-Lang and Holter has been altered to permit direct histological control and the use of smaller samples, and is not limited in applicability to the nervous system.
Abstract: The general histochemical procedure of Linderstrom-Lang and Holter has been altered to permit direct histological control and the use of smaller samples. Frozen sections are dried at –30°C. From the dry sections are cut out identified regions as small as 100 x 100 x 20µ (0.2γ wet weight). The chemical analysis of these fragments may be based on either their protein content, their dry weight or their volume. The weight and volume measurements are described and validated. The revised technique is not limited in applicability to the nervous system.
680 citations
TL;DR: A characteristic pattern of organization was found with the help of the electron microscope in sectioned animal mitochondria irrespective of the species providing the specimen and of the cell type examined.
Abstract: A characteristic pattern of organization was found with the help of the electron microscope in sectioned animal mitochondria irrespective of the species providing the specimen and of the cell type examined.Each mitochondrion was found to possess:1) A limiting membrane.2) A mitochondrial matrix that appears structureless at present levels of resolution.3) A system of internal ridges (cristae mitochondriales) that protrude from the inside surface of the membrane towards the interior of the organelles. In many mitochondria the cristae are perpendicular to the long axis of the organelles and occur in series within which they lie parallel to one another at more or less regular intervals.In favorable electron micrographs the mitochondrial membrane appears to be double and the cristae appear to be folds of a second, internal mitochondrial membrane.
591 citations
TL;DR: In many respects the nuclear and cytoplasmic detail revealed surpasses that which can be achieved by light microscopy and offers great promise for research in problems of cytophysiology and pathology.
Abstract: Technical methods for preparation of thin sections suitable for electron microscopy, while exacting, have been developed to a point of useful application. A series of electron micrographs from such...
183 citations
TL;DR: The very small mitochondria differ from the larger ones in the levels of activities of all enzymes studied, with the exception of adenosine-5’-phosphatase; the most striking differences were found in the cases of acid phosphatase and uricase.
Abstract: By separating the cytoplasmic granules of 0.88 M sucrose homogenates of rat liver into eight fractions it has been possible to demonstrate marked chemical and enzymatic heterogeneity among the isolated microsomes and less pronounced heterogeneity among the isolated mitochondria. The more readily sedimented microsomes are rich in ribose nucleic acid and show high esterase, adenosine-5’-phosphatase, acid phosphatase and uricase activities, while the less readily sedimented microsomes, although rich in ribose nucleic acid esterase and adenosine-5’-phosphatase have low levels of acid phosphatase and uricase activities. The very small mitochondria differ from the larger ones in the levels of activities of all enzymes studied, with the exception of adenosine-5’-phosphatase; the most striking differences were found in the cases of acid phosphatase and uricase.A centrifugation schedule is given to isolate a "nuclear fraction," a mitochondrial fraction, a mixed fraction of smallest mitochondria and microsomes (of ...
151 citations
TL;DR: Evidence and experimental results indicate that glucose-6-phosphatase is an enzyme distinct from nonspecific acid and alkaline phosphatases and that this enzyme can be so demonstrated histochemically.
Abstract: Glucose-6-phosphatase has been demonstrated in frozen sections of mouse liver and kidney. In both organs, the enzyme is present only within parenchymal elements and is absent from stromal tissues. The enzyme is more abundant in the peripheral third of the hepatic lobule than in the inner two-thirds. Within the hepatic cell the enzyme is concentrated about the nuclear membrane. Studies were made of the distribution of the enzyme in the liver in relation to the diurnal cycle but no significant changes in enzyme distribution were observed. In the kidney the enzyme is found in the proximal convoluted tubule, concentrated at the basal pole of the cells, and in portions of the renal glomerulus. The evidence and experimental results are described which indicate that glucose-6-phosphatase is an enzyme distinct from nonspecific acid and alkaline phosphatases and that this enzyme can be so demonstrated histochemically. The presence of this enzyme in the liver and kidney is correlated with the ability of these organ...
151 citations
TL;DR: α-Chloroacyl esters of α-naphthol and of naphthalol AS are hydrolyzed by mast cells and myeloid elements at a much faster rate than the corresponding regular acyl esters.
Abstract: α-Chloroacyl esters of α-naphthol and of naphthol AS are hydrolyzed by mast cells and myeloid elements at a much faster rate than the corresponding regular acyl esters. The enzyme responsible for t...
127 citations
TL;DR: The acquisition of acid phosphatase by monocytes during their transformation to macrophages appeared to be related to their enhanced phagocytic activity.
Abstract: Cultures of leukocytes from chicken blood grown in fluid medium in roller flasks were stained for lipids, carbohydrates, phosphatase, esterase, and succinic dehydrogenase in order to study the cytochemical changes associated with the transformation of monocytes to macrophages, epithelioid cells and multinucleate giant cells.The great variability in cell form observed by previous investigators in such cultures was confirmed. The cells were usually round but not uncommonly were stellate or fusiform, sometimes bearing a striking similarity to fibroblasts, at other times resembling sheets of mesothelium. The factors determining their size and shape were obscure, but low pH seemed to favor the formation of giant cells.Monocytes of chicken blood were ordinarily devoid of stainable lipid, carbohydrate and enzymes. Macrophages and epithelioid cells which developed from them were negative for esterase, succinic dehydrogenase and alkaline phosphatase but gave a strong reaction for acid phosphatase, localized in the...
121 citations
TL;DR: In conclusion, it appears that the reactivities of aldehyde-fuchsin and the Schiff reagent are similar in that they both show an affinity for alde Hyde groups, but different in that aldehyd-fucsin seems to possess an affinity to strong sulfur acids which the SchiffReagent does not.
Abstract: A comparison is made of the relative affinities of aldehyde-fuchsin and the Schiff reagent for various tissue elements in different states of oxidation.Glycogen, gastric mucus, kidney brush border, intestinal striated border, reticular tissue, gastric parietal cell canaliculi, and Paneth cell granules react with neither aldehyde-fuchsin nor the Schiff reagent without periodic acid or stronger oxidation.Beta cell granules of the islets of Langerhans, keratin, mast cells, elastic fibers, hyaline cartilage, goblet cells, thyroid colloid, argentaffine cell granules, and the acrosomes of spermatozoa are stained by aldehyde-fuchsin without oxidation. None of these tissue elements exhibit a clearly positive reaction with the Schiff reagent under similar conditions. However, elastic fibers, hyaline cartilage, and acrosomes do display a slight coloration.Since the tissue elements which do not stain with aldehyde-fuchsin without previous oxidation act in a similar manner toward both aldehyde-fuchsin and the Schiff ...
114 citations
TL;DR: It is shown that protein may modify the reaction between dye and polysaccharide by competing with the dye for the stainable groups and, since the competition shows a pH dependence, to an alteration of the “pH signature.”
Abstract: The basic dyes, particularly those which exhibit metachromasia, have been used extensively in histochemical studies of the connective tissue mucopolysaccharides. The specificity of the reaction depends upon the combination of dye with the acid groups of the polysaccharides. It is frequently assumed that this reaction occurs quantitatively. Thus in addition to the use of basic dyes to demonstrate the presence of mucopolysaccharides in tissues, the amount of dye bound under standard conditions has been considered to be a measure of the quantity of mucopolysaccharide present (1, 2). Furthermore the dye binding at different hydrogen ion concentrations has been interpreted as an indication of the dissociation characteristics of the acid groups of the stainable material and used to determine the so-called “pH-signature” of particular substances (3). It is the purpose of this paper to show that protein may modify the reaction between dye and polysaccharide by competing with the dye for the stainable groups. This may lead to a masking of the polysaccharide and, since the competition shows a pH dependence, to an alteration of the “pH signature.”
102 citations
TL;DR: The histochemical localization of esterases using α-naphthyl acetate and naphthol AS acetate as substrates is described for the five mammalian species, providing further evidence for the existence of an "esterase spectrum" consisting of a group of more or less specialized enzymes acting on certain specific esters of carboxylic acids.
Abstract: The histochemical localization of esterases using α-naphthyl acetate and naphthol AS acetate as substrates is described for the five mammalian species: man, rabbit, rat, cat, and mouse. The localiz...
94 citations
TL;DR: The histochemical localization of the enzyme was improved so as to enable the use of thin frozen sections (10 µ) by the addition of the following ions: bicarbonate, calcium, magnesium and aluminum.
Abstract: The distribution of succinic dehydrogenase activity was compared in the organs of six mammals. The histochemical localization of the enzyme was improved so as to enable the use of thin frozen sections (10 µ) by the addition of the following ions:bicarbonate, calcium, magnesium and aluminum. The distribution of enzymatic activity by this technic in the organs of the rat was compared with the earlier method.
TL;DR: In the course of this study, it was noted that the vast majority of the structures which have an affinity for aldehyde fuchsin are also chromotropic (metachromatic) and/or periodic acid-Schiff (PAS) positive after salivary digestion.
Abstract: The purple dye aldehyde fuchsin is obtained by adding paraldehyde to an acidified alcoholic solution of basic fuchsin (Gomori, i950b). In his original report Gomori recommended it primarily as an elastic tissue stain, but also mentioned its affinity for certain types of mucus, the chief cells of the fundic glands, and the cytoplasmic granules of some of the basophils of the anterior pituitary (the beta cells of Romeis), the beta cells of the islets of Langerhans and the mast cells. This remarkable selectivity of the dye induced us to investigate its staining properties more extensively. It was hoped that a survey of the structures which stain with aldehyde fuchsin might lay the groundwork for the understanding of the possible chemical basis of the method. In the course of this study it was noted that the vast majority of the structures which have an affinity for aldehyde fuchsin are also chromotropic (metachromatic) and/or periodic acid-Schiff (PAS) positive after salivary digestion. Consequently, in this report, the staining of a wide variety of mostly mammalian tissues with aldehyde fuchsin, toluidin blue and PAS will be compared. The effects of oxidation of the sections prior to staining with aldehyde fuchsin wifi also be discussed.
TL;DR: The sudanophilia of leucocytes depends on an as yet unexplained chemical combination of the dyes with cytoplasmic constituents which is probably not in the form of preexisting granules, rather than on lipoid staining, as in the Ciaccio method.
Abstract: It seems improbable that leucocyte oxidase is a fatty acid peroxide, as suggested by Sehrt, on these grounds:1. Positive reactions for aldehyde and for ethylene groups are not obtained. Hence the p...
TL;DR: The intestines, particularly the small intestine, were a possible excretory route for zinc and dithizone in aqueous-acetone solution alone demonstrated non-specific staining in stomach, prostate, pancreas, and erythrocytes and fat of dog, man, rabbit, and rat.
Abstract: A histochemical technique specific for zinc was achieved by means of a dithizone complex-forming solution. With this solution, zinc in prostate and stomach sections could be clearly differentiated from other metallic constituents. Dithizone in aqueous-acetone solution alone demonstrated non-specific staining in stomach, prostate, pancreas, and erythrocytes and fat of dog, man, rabbit, and rat. The staining of metals as contrasted with fat concentration of the dye was shown by loss of stain in tissues previously chelated with ethylenediaminetetracetic acid. Staining of phagocytized zinc oxide in macrophages indicated penetration of the cellular membranes by dithizone. Zinc acetate given intravenously to rabbits showed a marked concentration of zinc in the epithelial cells of small and large intestine. This experiment indicated that the intestines, particularly the small intestine, were a possible excretory route for zinc.
TL;DR: The ability to recover large fractions of the total enzyme activity in isolated mitochondria and to demonstrate concentration of this activity in mitochondria indicated to us that the findings were accurate from the cytochemical standpoint.
Abstract: Almost 20 years have passed since the isolation of mitochondria from guinea pig liver was announced by Bensley and Hoerr (6) in 1934. This important discovery remained largely unexploited until the researches of Claude on the particulate components of cytoplasm were begun some 5-6 years later. Although Claude at first mistook the mitochondria for secretion granules (17), his methods for the isolation of these cell organelles, which were the first to be described in the literature (17, 19, 20), laid the foundations for the large amount of research on mitochondria that has been carried out in the last few years. The recognition of the key role played by the mitochondrion in the metabolism of the cell is largely the result of the localisation of a number of important enzymatic functions in isolated mitochondria (cf. 97). These enzymatic activities have not merely been detected in isolated mitochondria but, in many cases, have been shown to account for 50 per cent or more of the total cellular activity. The ability to recover large fractions of the total enzyme activity in isolated mitochondria and to demonstrate concentration of this activity in mitochondria indicated to us that the findings were accurate from the cytochemical standpoint. On the other hand, we have hesitated to attach cytochemical significance to the presence of small percentages of the total tissue enzyme activity in mitochondna or in any other subcellular structure, since such results could very readily be the result of adsorption, contamination, or other factors which could not be satisfactorily proved or disproved. Even in those cases in which it has been possible to demonstrate almost exclusive localisation of a cellular function in mitochondria, the possibility of artifacts has always been considered and we have sought and welcomed the appearance of independent methods that would either support or deny the results obtained with isolated mitochondria. Although some such evidence has been reported in support of the localisation of certain enzymes in mitochondria (3, 11), confirmation of the results of the cell fractionation technic has not kept pace with new results obtained with this method. It is hoped that this situation will be corrected in the future.
TL;DR: A systematic study of the enzyme systems which are capable of reacting with Janus green B and its reduction products indicates that this dye is reduced by the lactic dehydrogenase enzyme system and suggest that JanusGreen B staining is dependent upon the enzyme activities of the cell.
Abstract: In 1900, Michaelis used Janus green B as a supravital stain (20). He observed that filamentous structures within the cytoplasm of the cell stained with this dye and he recognized that these structures were similar to the elementary filaments described by Altmann in 1890 (1). Michaeis had specified that the diethyl derivative of Janus green was necessary for supravital staining. However, a number of subsequent investigators were unsuccessful in using this dye as a supravital stain, presumably because they did not use the diethyl derivative (10). Bensley reintroduced the use of Janus green B as a supravital stain (4). He was careful to use the diethyl derivative and he showed that the structures which stained with Janus green B were identical with the structures which Benda called mitochondria (3). The Janus green B staining reaction is oxygen dependent (5, 6), for although selective staining of mitochondria appears under partial anaerobic conditions, these structures become decolorized when all of the oxygen is removed. The Janus green B staining reaction is reversibly inhibited by cyanide (18, 19); when cyanide is added to supravitally stained mitochondria they become colorless whereas when the cyanide is subsequently removed by washing, the mitochondna regain their blue color. In vitro studies with Janus green B have indicated that this dye is reduced by the lactic dehydrogenase enzyme system (2). All of these observations suggest that Janus green B staining is dependent upon the enzyme activities of the cell. In order to clarify the enzymatic mechanism involved in the Janus green B staining reaction, we have undertaken a systematic study of the enzyme systems which are capable of reacting with Janus green B and its reduction products. Several preliminary notes have been published (7, 14, 15) and three completed papers are in press (16, 8, 9). Further studies on the reduction of Janus green B by the isolated cell components and on the binding of Janus green B to proteins and to isolated cell components are in progress and will be published soon (17).
TL;DR: Ferric ferricyanide mixtures to Prussian blue and most of the other substances tested reacted feebly or slowly, and probably need not be considered in the histochemical test, provided the reaction time is kept down to 10 to 15 minutes.
Abstract: Ascorbic, oxalic and uric acids, phenols, indols, pyrrols, aryl amines, hydrazines, thiols and inorganic sulfides and peroxide promptly reduce ferric ferricyanide mixtures to Prussian blue. Most of the other substances tested reacted feebly or slowly, and probably need not be considered in the histochemical test, provided the reaction time is kept down to 10 to 15 minutes.
TL;DR: The life history of mitochondria remained obscure however because of the cytologists’ inability to distinguish between these bodies and the cytoplasmic particles of similar size which are destined to develop into the various types of plastids characteristic of the plant cell.
Abstract: existence of mitochondria in plant cells was first described by Meves (1904) who observed them in the pollen nurse cells of the Nymphaeceae. The presence of mitochondria in plant cells was subsequently confirmed by a number of investigators. The life history of mitochondria remained obscure however because of the cytologists’ inability to distinguish between these bodies and the cytoplasmic particles of similar size which are destined to develop into the various types of plastids characteristic of the plant cell. Sorokin first (1938, 1941) achieved such a differentiation when she found that it is possible to differentiate mitochondria from morphologically similar forms of plastids by vital staining of the mitochondria with the dye Janus green B.
TL;DR: Evidence is presented that the abolition of the staining of a polysaccharide in tissue sections can be produced by means other than that of specific hydrolysis.
Abstract: The limitations of the staining reactions for polysaccharide constituents of tissues makes necessary the use of other techniques. Enzymatic digestion of stainable material in tissue sections is now frequently used as a means of identification (1 , 2, 3, 4, 6). The use of such methods involves the following assumption: Absence of staining following application of an enzyme preparation containing demonstrable polysaccharide-splitting activity indicates the presence of specific enzyme substrate. If pure enzyme preparations with all activities specifically known were available it would be possible under appropriate conditions to make this inference. In the absence of such enzymes only crude or partially purified preparations can be used for purposes of defining these tissue constituents. It is the purpose of this paper to present evidence that the abolition of the staining of a polysaccharide in tissue sections can be produced by means other than that of specific hydrolysis. Consequently the results following the use of such preparations must be interpreted with considerable caution.
TL;DR: The properties of these cytoplasmic granules in bacteria are summarized, in so far as they have been worked out, and thus to arrive at a definition of the mitochondria of bacteria.
Abstract: Recent evidence from the biochemical, the genetic and the morphologic study of bacteria, in that chronological order, has indicated essential similarities of the bacterial cell to the cells of higher organisms. Recognition in bacteria of a large category of cytoplasmic granules as possessing characteristics which strongly suggest that they are the functional equivalents of the mitochondria of anirnaE and plant cells, has been a significant step. It is the purpose of this communication to summarize the properties of these cytoplasmic granules in bacteria, in so far as they have been worked out, and thus to arrive at a definition of the mitochondria of bacteria. Morphology. In electron micrographs under conditions in which the surround-
TL;DR: Various supravital mitochondrial indicators have been used on three strains of S. typhosa, and the mitochondria were differentiated from nuclei and cell wall, and showed no structural relationship to terminal flagellar spheres.
Abstract: Evidence for the existence of mitochondria in bacteria has been presented in papers by Mudd, Wititerscheid, DeLamater and Henderson (1951) ; Mudid!, Brodie, Winterscheid, Hartman, Beutner atid McLean (1951) ; atsd Witsterscheid and Mudd (1953). The present work is concerned with the further study of bacterial mitochondria by means of cytological examination alsdl comparison of one tionmotile arid two motile strains of Salmonella typhosa with the aid of mitochotidrial, lipid, nuclear, fiagellar and! cell wall staitis. No straiti differences in mitochotidria were observed with these orgatsisms. Therefore, all descriptions may he considered applicable to each of the three strains used! utsless otherwise indicated.
TL;DR: Ca phosphate, when precipitated by the addition of phosphate to solutions similar in composition to the histochemical substrate mixture, does not show any experimentally demonstrable tendency to supersaturation.
Abstract: Ca phosphate, when precipitated by the addition of phosphate to solutions similar in composition to the histochemical substrate mixture, does not show any experimentally demonstrable tendency to supersaturation. The factors which may contribute to false localizations of enzymatic activity its the Ca-CoS method for alkaline phosphatase are not well understood and certainly not amenable to mathematical analysis for the time being. At present, diffusion artifacts do not appear to be an important source of error, provided incubation time is not unduly extended.
TL;DR: The positive coupling with diazo-safranin speaks against a catechol or hydroquinone structure and supports the thesis of a resorcinol structure.
Abstract: Freshly diazotized safranin O colors enterochromaffin cell granules black. Background staining can be kept down to a fairly light red. Fairly stable acid aqueous safranin solution and normal sodium nitrite solution can be kept on hand and mixed ex tempore to prepare the diazo solution. A diazotization time of 15 minutes at 3°C is adequate, and the coupling (staining) time of 5 minutes should not he exceeded. Both the specific and the background stains are acid fast. A 1:40 dilution in M/10 Na2HPO4 produces the desired grade of alkalinity and a proper dilution for best. contrast.In vitro alkaline coupling reactions with diazo-safranin produced colored precipitates with phenol, cresols, naphthols, resorcinol, hydroquinone, pyrogallol and tyrosine. Definite colors were produced on gelatin paper impregnated with resorcinol and the naphthols, less definite with phenol and the cresols. Gelatin paper models gave positive reactions with diazotized α-naphthylamine for catechol, pyrogallol and phloroglucinol as wel...
TL;DR: The alkaline tetrazolium reaction is considered to form the basis of a useful method both for research into the process of keratinisation and also for the routine diagnosis of abnormal tumour keratins.
Abstract: 1) Hydrolysis of combined cystine at pH 12.8 gives rise to reaction products capable of converting soluble, relatively colourless, tetrazolium salts into insoluble formazan dyes.2) Both normal and abnormal keratins are demonstrated by the reaction.3) Cystine-free structures which also reduce tetrazolium salts at pH 12.8 are present in tissue sections. These are either lipid-containing or reducing sugar-containing, but distinction between these two groups and the cystine-containing group of structures is histochemically possible.4) The alkaline tetrazolium reaction is considered to form the basis of a useful method both for research into the process of keratinisation and also for the routine diagnosis of abnormal tumour keratins.
TL;DR: In order to demonstrate potassium in tissues it was necessary to develop a reliable staining technique and it became apparent that failures in the past were due to the rapid shift of potassium across cell membranes.
Abstract: synthesis of glycogen from glucose is greater in media high in potassium (2, 7). The shift of glucose accompanied by potassium from the blood into the liver in treated diabetics has been well documented (8). In contrast, in vitro studies using skeletal muscle and heart have suggested that glycogen synthesis is enhanced by media low in cation content (19, 20). The indispensability of potassium in the trans-phorylation of phosphopyruvate with adenosine tnphosphate has been shown (1) and confirmed (16). Pathologic deposits of glycogen accumulate around the borders of myocardial infarcts (21), particularly along the suhendocardium.1 During the past two years we have attempted to correlate the distribution of glycogen and potassium in these areas adjacent to myocardial infarcts in view of the possibility that zonal alterations of potassium and glycogen might be a physioanatomic basis for the occurrence of dardia( arrythmias in coronary artery disease (myocardial fibrosis, infarction). The important role of potassium in iteuro-muscular conduction and the location of specialized conduction pathways, rich in glycogen, in the immediate suhendocardium suggested that redistribution of these substances under the stress of anoxia might result in the development of aberrant conduction pathways or retrogressive changes in preexisting routes. In order to demonstrate potassium in tissues it was necessary to develop a reliable staining technique. Accordingly, previously reported methods were reviewed and critically evaluated. It became apparent that failures in the past were due to the rapid shift of potassium across cell membranes. Certain modifications were devised to minimize this shift.
TL;DR: It is shown that the same dye complex was formed in acid-fast cells as in the test-tube when the absorption spectra were determined for materials in tissue stained with crystal violet.
Abstract: Recent work from this laboratory (Berg, 1) has shown that the acid-fastness seen in mycobacteria can reside in two different properties of the cells. First, there was an acid-fastness dependent upon cell structure and shown by intact but not by crushed tubercie bacilli. Secondly, in the same cells there was an acid-fastness not altered by structural changes but rather dependent upon chemical properties. It was lost when and only when mycolic acid was removed from the cells. Further, purified mycolic acid showed the same type of acid-fastness and to the same degree. Lepra bacilli and sperm showed only the second type of acid-fastness and again it was also found in the mycolic-acid-like lipids extracted from these cells (Berg, 1, 2). Further work (Berg, 3) described the unique reaction that took place between these lipids and one dye which was capable of giving the acid-fast reaction, crystal violet. Among other properties, the dye-mycolic acid complex exhibited intense absorption of light at 3500 A while such absorption was not shown by the dye alone, the acid alone, or any other type of acid tested in combination with the dye. In this paper, it is shown that the same dye complex was formed in acid-fast cells as in the test-tube when the absorption spectra were determined for materials in tissue stained with crystal violet. In addition, a number of other reactions of the acid-fast material in the cells are described. The separation of the acid-fastness of mycobacteria from that shown by keratohyaline granules, Russell bodies and ceroid gives at least this first type of acid-fastness the character of a histochemical test.
TL;DR: Prolonged exposure to postassium bichromate solution creates weak diffuse aldehyde reactions in tissues, and also tends to weaken the reactivity of collagen to the periodic acid Schiff procedure.
Abstract: The use of alcoholic solutions of periodic acid does not greatly impair the periodic acid Schiff reaction of collagenThe Hotchkiss acidulated thiosulfate "reducing rinse" is essentially a sulfurou
TL;DR: The results of treatment of the sections with halogens, oxidants and reductants suggest that the aldehydic substances which react with the Schiff reagent are all derived from unsaturated lipids but that, in addition, ketonic substances are formed or unmasked which are reactive to the hydrazide alone.
Abstract: Fresh-frozen sections of mouse ovary, briefly fixed in formalin, fail to stain with fuchsin-sulfurous acid (the Schiff reagent) or the Ashbel-Seligman carbonyl reagents. When the sections are treated for 10 minutes with mercuric chloride and then stained, the cytoplasm of the cells acquires a faint coloration—the plasmal reaction. When the sections are washed in water or dilute alcohol for 3 or 24 hours, however, lipid droplets become increasingly stainable. The results of treatment of the sections with halogens, oxidants and reductants suggest that the aldehydic substances which react with the Schiff reagent are all derived from unsaturated lipids but that, in addition, ketonic substances are formed or unmasked which are reactive to the hydrazide alone. Whether or not the ketonic lipids are steroidal has not been demonstrated.
TL;DR: It is demonstrated that the interpretation of the histochemical dopa reaction, discovered by Bloch, correlated with mammalian melanogenesis in vivo has been greatly influenced by the analogy of the tryosinase-tyrosin reaction in vitro with the same process.
Abstract: 1. It is demonstrated that the interpretation of the histochemical dopa reaction, discovered by Bloch, correlated with mammalian melanogenesis in vivo has been greatly influenced by the analogy of the tryosinase-tyrosin reaction in vitro with the same process.2. Repeating Bloch's histochemical pretreatment experiments on fixed tissue proved that the extreme lability found by Bloch must be attributed to detachment of the factor from the section or insufficient control of pH of the pretreatment solution.3. Sufficient lability remains to be in line with an enzymatic nature, though classifying the factor in the realm of copper containing enzymes must be considered premature for lack of comparable data in the extract enzymological field.
TL;DR: The phosphamidase content in normal and pathologic tissues of 37 biopsies from the oral cavity was examined using the histochemical method of Gomori, finding high concentrations in the epithelia of epidermoid carcinoma, leukoplakia, papilloma, pleomorphic salivary gland tumor and in the proliferating connective tissue of a giant cell tumor.
Abstract: The phosphamidase content in normal and pathologic tissues of 37 biopsies from the oral cavity was examined using the histochemical method of Gomori.High concentrations of phosphamidase were found in the epithelia of epidermoid carcinoma, leukoplakia, papilloma, pleomorphic salivary gland tumor, in the proliferating connective tissue of a giant cell tumor and in the epithelium adjacent to a fibroma and in the epithelium of blastomycosis. Low concentrations of phosphamidase were found in a few cases of inflammation of the mucous membrane associated with hyperkeratosis.The phosphamidase reaction was negative for normal epithelium and connective tissue of the oral mucous membrane, the lining of the maxillary sinus and mucous glands.The present limitations of the technique are discussed and some precautions against erroneous conclusions presented.