bluuebunny/crossref_metadata_2025 huggingface dataset:
ENZYME LEVELS OF INDIVIDUAL NEURONS IN RELATION TO LIPOFUSCIN CONTENT
Authors: HIRSCH HILDE E.
Published: April 2011

By quantitative histochemical methods, activities of (α-naphthyl) acid phosphatase and β-galactosidase, and of malate and lactate dehydrogenase, were measured in individual human anterior horn nerve cell bodies selected for high and low lipofuscin content. No significant differences were found. However, when single neurons were subdivided into pigmented and unpigmented portions, the two lysosomal enzymes were considerably more active, and malate and lactate dehydrogenase less active, in the lipofuscin-rich part of the cell. These results are consistent with the view that lipofuscin granules are derived from lysosomes.
MICROSPECTROPHOTOMETRY OF NEWLY SYNTHESIZED STARCH IN SITU
Authors: WHEELER GEORGE E.
Published: April 2011

Many of the cells in stem sections of several Commelinaceae species synthesized much new starch when incubated in buffered 1% glucose 1-phosphate solution. The starch appeared in the cytoplasm rather than in the plastids. Although the starch I2-KI color was uniform within any one cell, there was considerable variation from cell to cell, even in the same section. The colors with I2-KI ranged from blue, through purples to magenta and mahogany. Tests with α-amylase and with β-amylase showed the starch to be amylose. Microspectrophotometrically determined extinction curves, based on the new starch in situ, varied with the visualized color. As expected, starch which stained blue with I2-KI had an absorption maximum in the orange-red wavelengths above 600 mµ; increasingly red I2-KI colors were characterized by shifts of the absorption maximum further into the shorter wavelengths. The course of new starch digestion by α-amylase and by β-amylase was followed visually and with the microspectrophotometer. Similarities and differences between these spectral curves and those published for in vitro studies are pointed out. The difficulties met with in using the microspectrophotometric method are discussed.
PERCHLORIC ACID EXTRACTION OF DEOXYRIBONUCLEIC ACID FROM THIN SECTIONS OF EPON-ARALDITE-EMBEDDED MATERIAL
Authors: DOUGLAS WILLIAM H. J.
Published: April 2011

A method is presented for the extraction of deoxyribonucleic acid from thin sections of tissue fixed in glutaraldehyde and osmium tetroxide and embedded in water-insoluble epoxy resin. Incubation of thin sections in 20% perchloric acid at 60°C for 8 hr produced electron-transparent bands in the polytene chromosomes from Drosophila salivary glands. Chromosome bands were also rendered electron-transparent when hydroxypropylmethacrylate-embedded thin sections were digested with deoxyribonuclease or extracted with perchloric acid. Nucleolar ribonucleic acid was unaffected by the perchloric acid extraction.
THE SPECIFICITY OF THE PYROANTIMONATE TECHNIQUE TO DEMONSTRATE SODIUM
Authors: TORACK RICHARD M. , LAVALLE MARIA
Published: April 2011

The use of an aqueous solution of pyroantimonate as a specific histochemical marker for intracellular sodium is not supported by this study since, at physiologic levels, potassium as well as sodium appears to be precipitated by such a pyroantimonate solution. Furthermore, a low pH and the addition of OsO4 as a fixative compound appear to favor the production of nonspecific electron-dense precipitates with this technique in vitro and in vivo. The addition of phosphate anion to a solution of pyroantimonate seems to utilize the sites at which many cations bind to pyroantimonate, thus elevating the concentration of cation needed to produce a precipitate. Artificial elevation of sodium above physiologic levels results in a precipitate with phosphate-buffered pyroantimonate and probably represents a valid demonstration of sodium since no other cation is present in sufficient concentration to produce an electron-dense deposit with this medium.
HUMAN AMYLOID PROTEIN: CHEMICAL VARIABILITY AND HOMOGENEITY
Authors: HARADA M. , ISERSKY C. , CUATRECASAS P. , PAGE D. , BLADEN H. A. , EANES E. D. , KEISER H. R. , GLENNER G. G.
Published: April 2011

The morphology of the fibril of amyloid derived from different individuals is similar, but occasionally significant differences are noted. All human amyloid filaments have a "β-pleated sheet" conformation as revealed by x-ray diffraction, and those examined after orientation show a "cross-β" pattern. All amyloid fibril concentrates studied so far can be fractionated to obtain the major amyloid protein component(s) by sequential gel filtration with 5 M guanidine-HCl in 1 N acetic acid on Sepharose 4B and Sephadex G-100 or G-75 columns with the removal of over 28% of proteins representing minor constituents. The major amyloid protein(s) obtained from the spleen and/or liver of six patients is found to contain tryptophan, to be deficient in hydroxylysine and hydroxyproline and usually at least one commonly occurring amino acid and to have a high content of dicarboxylic acid and short chain amino acids and unreactive (blocked) NH2-terminal groups or aspartic acid-asparagine (Asx). However, the amyloid protein(s) from each individual differs from that of the others in molecular weight, in amino acid composition and in the presence or absence of specific tryptic peptides. Amyloid protein(s) from the liver and spleen of the same individual is identical. No chemical characteristics distinguish amyloid proteins derived from cases classified clinically as "primary" from those classified as "secondary." There is a striking chemical similarity between amyloid proteins and the NH2-terminal variable fragment of the light and heavy chain of immumoglobulin proteins.
SMALL CYTOPLASMIC BODIES IN THE LOOP OF HENLE AND DISTAL CONVOLUTED TUBULE THAT RESEMBLE PEROXISOMES
Authors: CHANG CHUNG-HO , SCHILLER BERNICE , GOLDFISCHER SIDNEY
Published: April 2011

Renal peroxisomes are believed to be restricted to the proximal convoluted tubules. However, we have found particles in the loop of Henle and distal convoluted tubule that resemble the peroxisomes of the proximal tubules in some respects. Although smaller in size (generally too small to be resolved in the light microscope), they have a finely granular homogenous matrix and are delimited by a single unit membrane. Whether these are microbodies (peroxisomes) or dense bodies (lysosomes) cannot be determined solely on the basis of their fine structure. These bodies do not show acid phosphatase activity, indicating that they are not lysosomes. When incubated in a diaminobenzidine medium that demonstrates the peroxidatic activity of the peroxisomal enzyme catalase, the small bodies are reactive, suggesting that they are peroxisomes. This activity is blocked by 3-amino-1,2,4-triazole, an inhibitor of catalase.
VARIATIONS IN THE LOCATION AND SIZE OF PYROANTIMONATE PRECIPITATES IN THE IMMATURE RAT CEREBRAL CORTEX
Authors: SUMI S. M.
Published: April 2011

The extracellular space in the developing rat cerebral cortex is quite large but varies considerably in extent depending on age, method of fixation and osmolarity of the fixative. Since Na+ is known to be extracellular in location, the potassium pyroantimonate technique of Komnick was utilized in the newborn rat in order to follow the movement of Na+ and water. However, the location and size of the pyroantimonate precipitates appeared to vary with the type of fixative solution and method of fixation. Thus, they were large and mainly intracellular with phosphate- or s-collidine-buffered glutaraldehyde after perfusion fixation, and mostly extracellular after immersion fixation. Following primary osmium tetroxide fixation, these particles were small and numerous with the aqueous solution, being diffusely present throughout the section, and large and fewer with the phosphate-buffered solution. Whether Na+ alone was being demonstrated could not be determined. Because of this marked variability it is concluded that definite statements as to the actual in vivo location of the Na+ in this preparation cannot be made.
AN IMMUNOFLUORESCENT STUDY OF ANTIGENIC STRUCTURES IN THE DEVELOPMENT OF HUMAN EPIDERMIS
Authors: DELPECH-STEWART ANNIE
Published: April 2011

The time of appearance of antigenic structures that react with circulating antibodies occurring in patients with bullous dermatosis was studied in embryonic skin by an immunofluorescent method. It was found that these structures are present in the 4th month of fetal life.
ON PHOSPHOTUNGSTIC ACID STAINING. II
Authors: QUINTARELLI G. , CIFONELLI J. A. , ZITO R.
Published: April 2011

Phosphotungstic acid (PTA) used at different concentrations in water was combined with various compounds (amino acids, proteins, acid glycosaminoglycans, glycoproteins and simple sugars). The results obtained indicated that PTA interacts with positively charged groups, that the sugar-hydroxyls do not take part in the interaction and that the type of binding involves electrostatic forces. To substantiate these findings further, heparin and heparin oligosaccharides were N-desulfated, combined with PTA and the precipitates were analyzed. It was found that the interaction had taken place only between the metal and the desulfated hexosamine amino groups and that when the N-desulfated product was acetylated no PTA binding occurred. When PTA was used at higher concentrations and in extremely acidic conditions, a precipitation of proteins, amino acids, acid glycosaminoglycans and neutral sugars ensued. This complex formation was interpreted as due to ionic interaction between the polyacid and the protonated hydroxyl groups.
THE EFFECT OF SAPONIFICATION UPON CERTAIN HISTOCHEMICAL REACTIONS OF THE EPITHELIAL MUCINS OF THE GASTROINTESTINAL TRACT
Authors: CULLING C. F. A. , REID P. E. , DUNN W. L.
Published: April 2011

An increase in periodic acid-Schiff reactivity after saponification has been demonstrated in the gastrointestinal tract mucins of man, rat, rabbit and guinea pig. Tins pH-dependent phenomenon is independent of fixation and processing since it occurred in cryostat sections of fresh and formalin-fixed tissue and in paraffin sections of tissues fixed in eight different fixatives. In man and rat this effect occurs at the level of the ileocecal valve and below; in guinea pig it is confined to the large intestine and rectum; while in rabbits it occurs in Brunner's glands, the ileocecal valve and below. In man the increase is found throughout the crypts whereas in rabbits the effect is seen mainly at the luminal end; in rats it is seen largely at the base of the crypts; and in guinea pigs it is variable. This effect, due to an increase in the number of 1:2 glycol groups, is accompanied in man, rat and guinea pig by an increase in basophilia due to the presence of sulfate and/or other highly acidic groups.
OBSERVATIONS ON PEROXISOMES IN BROWN ADIPOSE TISSUE OF THE RAT
Authors: AHLABO IRÉNE , BARNARD TUDOR
Published: April 2011

During cytochemical studies of brown adipose tissue from rat, cytoplasmic organelles that apparently show peroxidative activity have been observed. The majority of the organelles have a diameter of 0.1-0.8 µ and a finely granular homogeneous matrix and are delimited by a single unit membrane. No sign of a "crystalloid" was seen. In order to demonstrate the peroxidative activity of the peroxisomal enzyme catalase in the organelles, brown adipose tissue was incubated in a medium containing 3,3'-diaminobenzidine tetrahydrochloride, after prefixation in 3% glutaraldehyde. The activity was blocked by 3-amino-l,2,4-triazole (an inhibitor of catalase) but not by KCN. Omission of exogenous hydrogen peroxide did not inhibit the reaction in the organelles. It is concluded that rat brown adipose tissue contains peroxisomes and, since the abundance of these organelles varies according to the physiologic activity of the tissue, peroxisomes may have a role in the thermogenic metabolism of brown fat.
INHIBITION OF PEROXIDASE BY METHANOL AND BY METHANOL-NITROFERRICYANIDE FOR USE IN IMMUNOPEROXIDASE PROCEDURES
Authors: STRAUS WERNER
Published: April 2011

The cytochemical reaction for peroxidase is partially inhibited by methanol. The addition of a small amount of sodium nitroferricyanide to the absolute methanol causes further inhibition. The activity of peroxidase is completely suppressed by methanol containing 1% sodium nitroferricyanide and l% acetic acid. The antibody to horseradish peroxidase is much less affected by these inhibitors than horseradish peroxidase itself. By using appropriate combinations of these inhibitors and by varying the time or temperature of treatment, the activity of injected horseradish peroxidase, or of endogenous peroxidase in leukocytes, erythrocytes or normoblasts, can be much decreased or suppressed while the specific ability of the antibody to adsorb horseradish peroxidase can be preserved to a considerable extent. KCN and NaN3 inhibit peroxidase activity only moderately under conditions required for immunoperoxidase procedures, i.e., after removal of the inhibitors.
A CENTRIFUGATION TECHNIQUE FOR CYTOCHEMICAL PREPARATIONS
Authors: LEEMANN U. , WEISS S. , SCHMUTZ E.
Published: April 2011

A centrifugation technique for microscopical preparations of cells or cell particles either unfixed or previously fixed in suspension has been developed. It allows more consistent results of cytofluorometric and interferometric measurements than the conventional smear technique.
THE FINE STRUCTURAL LOCALIZATION OF ACETYLCHOLINESTERASE ACTIVITY IN THE RETINA AND OPTIC NERVE OF RABBITS
Authors: REALE E. , LUCIANO L. , SPITZNAS M.
Published: April 2011

In the rabbit retina acetylcholinesterase activity is localized in the perinuclear cisterna, in the cisternae of the rough surfaced endoplasmic reticulum and in the Golgi apparatus of ganglion cells and amacrine cells. The histochemical reaction is positive also in the rough surfaced endoplasmic reticulum of some horizontal cells. The highest activity is seen in the internal plexiform layer; because of artifacts caused by the diffusion of the enzyme, a clear demonstration of relation of the positivity to one or the other regular components of this layer, however, is not possible. Myelinated fibers which exhibit acetylcholinesterase activity and are most probably efferent are found in the internal plexiform layer. In the retinal nerve fiber layer and in the optic nerve only a few fibers show a positive reaction.
LOCALIZATION OF ACETYLCHOLINESTERASE VIA PRODUCTION OF OSMIOPHILIC POLYMERS: NEW BENZENEDIAZONIUM SALTS WITH THIOLACETATE FUNCTIONS
Authors: MEDNICK MORTON L. , PETRALI JOHN P. , THOMAS NORMAN C. , STERNBERGER LUDWIG A. , PLAPINGER ROBERT E. , DAVIS DONALD A. , WASSERKRUG HANNAH L. , SELIGMAN ARNOLD M.
Published: April 2011

By incorporating an enzyme-susceptible thiolester group and a diazonium group into the same molecule, it has been possible to obtain an osmiophilic polymer upon enzymatic hydrolysis of the thiolester, via diazothioether formation linking the units. The thiolacetates with diazonium groups are specific substrates for cholinesterase because of the strong positive charge on the diazonium group. Following osmication, sites of acetylcholinesterase in motor end plate are notable as black deposits in light microscopy and opaque deposits in electron microscopy. This is the first cytochemical demonstration of a hydrolytic enzyme using a single agent as both substrate and capture reagent to form a polymer. The insolubility of the polymer in both water and lipid before osmication provides precision of localization needed for the high resolution of electron microscopy. However, the effectiveness of the new principle for electron microscopy depends as well upon the rate of the capture reaction (polymerization), which in turn depends upon the pK a of the thiol and pH and temperature at which the cytochemical reaction is conducted.
DEMONSTRATION OF ACID PHOSPHATASE ACTIVITY USING 1-ACETYL-3-INDOLYL PHOSPHATE AS SUBSTRATE
Authors: HAYASHI MASANDO
Published: April 2011

A simultaneous coupling azo-indoxyl method for the cytochemical demonstration of acid phosphatase activity using p-toluidine salt of 1-acetyl-3-indolyl phosphate is described. A satisfactory staining for the enzyme activity was obtained following incubation of formol-calcium-fixed frozen sections for 30 min at 25°C in a medium containing 1 mM each of the substrate and hexazonium pararosanilin and adjusted to pH 4.5-5.0 with acetate buffer. The distribution of acid phosphatase activity demonstrated by this method was identical with that obtained either by Gomori's technique using β-glycerophosphate as substrate or by the Barka and Anderson's naphthol AS-BI phosphate-hexazonium pararosanilin method in several tissues of male rats so far examined. However, the adrenal enzyme activity was most prominent in the medulla with 1-acetyl-3-indolyl phosphate and β-glycerophosphate but it was more marked in the cortex with naphthol AS-BI phosphate. An advantage of using 1-acetyl-3-indolyl phosphate as substrate is that the same compound can be used for comparing azo-indoxyl and lead-salt methods. Effects of phospholipase C and Triton X-100 on staining for acid phosphatase were tested by pretreating fixed rat liver and kidney sections with these agents and incubating them in the medium containing 1-acetyl-3-indolyl phosphate and either hexazonium pararosanilin or lead ions as a coupler. The pretreatment did not change discrete lysosomal staining, as seen in untreated controls, using pararosanilin as a coupler, but greatly modified the staining using lead ions. The results indicate that the preciseness of staining for acid phosphatase with lead-salt method is highly dependent on some lipid material which attracts lead in tissues and that appropriately devised azo dye or azo-indoxyl methods demonstrate enzyme sites more accurately than lead-salt method.
ALTERATION OF HISTONES FROM MOUSE EPIDERMAL CELLS AFTER INCUBATION WITH ELASTASE AND HYALURONIDASE
Authors: SEGAL ALVIN , SCHROEDER MARGARET , VAN DUUREN BENJAMIN L.
Published: April 2011

Chromatin was isolated from whole mouse skin, mouse epidermal cells and mouse liver by standard procedures used for isolation of chromatin from other mammalian tissues. Chromatin from whole mouse skin or from mouse epidermal cells had not been isolated or characterized earlier. For the preparation of chromatin from mouse epidermal cells, the latter was separated from dermis by incubation for 30 min at 37°C in a solution containing the enzymes elastase and hyaluronidase. The relative proportions of the chromatin components, the T m and the ultraviolet absorption spectrum were all similar to that of chromatin from whole mouse skin which was not treated with enzymes and to other mammalian chromatin preparations. Electrophoresis of the histones from epidermal chromatin in polyacrylamide gels revealed the absence of histones F1, F3 and F2a2 and the appearance of a new band. Histones isolated from chromatin prepared from the whole mouse skin had a gel electrophoresis pattern virtually identical with histones isolated from mouse liver chromatin and to reported histone patterns from other mammalian tissues. The alterations in mouse epidermal histones are similar to reported changes in histones from calf thymus nucleohistone previously subjected to incubation at various temperatures. The enzymatic incubation technique can therefore not be used as a method of isolating unaltered mouse epidermal chromatin. The findings illustrate that very subtle chemical alterations can be induced by usual methods of tissue preparation and that these changes can only be detected by highly sensitive analytical techniques.
DEGRADATION OF GLUCOSE 6-PHOSPHATE DEHYDROGENASE INTO ENZYMATICALLY ACTIVE MOLECULAR FORMS BY TRYPSIN
Authors: HORI SAMUEL H. , NODA SETSUKO
Published: April 2011

One of the hepatic glucose 6-phosphate dehydrogenase isozymes of rats, designated "C," has been degraded by tryptic digestion into two enzymatically active forms of different electrophoretic mobility. The modified forms exhibited the same substrate specificity and mobility as the isozymes previously designated E and E'. The results of the present study emphasized that zymograms of tissues rich in proteinases should be interpreted with caution.
NUCLEOSIDE DIPHOSPHATASE AND THIAMINE PYROPHOSPHATASE ACTIVITIES IN THE ENDOPLASMIC RETICULUM AND GOLGI APPARATUS
Authors: GOLDFISCHER SIDNEY , SCHILLER BERNICE , ESSNER EDWARD
Published: April 2011

The effects of pH, fixatives and divalent ions on nucleoside diphosphatase (NDPase) and thiamine pyrophosphatase (TPPase) activities in the endoplasmic reticulum (ER) and Golgi apparatus (GA) were examined in adult and neonatal hepatocytes and other cell types in the rat. In liver cells TPPase and NDPase both have a similar localization in the rough ER, nuclear envelope and smooth ER but differ in their pH optima; TPPase is most active at pH 8, NDPase at pH 7. TPPase in the GA, unlike its counterpart in the ER, is most active at neutral pH. High levels of NDPase activity are present in the GA of neurons, epididymis and other cells, but not in hepatocytes. TPPase in the ER, but not the GA, is stimulated by the addition of adenosine triphosphate to the medium. These observations show that different conditions are required to demonstrate ER and GA diphosphatase activities. Whether separate enzymes or multiple configurations of a single protein are responsible for these activities cannot be determined by staining procedures.
GLUCOSE OXIDASE AS AN ANTIGEN MARKER FOR LIGHT AND ELECTRON MICROSCOPIC STUDIES
Authors: KUHLMANN W. D. , AVRAMEAS S.
Published: April 2011

A new staining procedure for glucose oxidase at both the light and the electron microscopic level has been developed. The procedure was equally effective for the detection of galactose oxidase. It appears that the method can be applied to all oxidoreductases which catalyze the production of hydrogen peroxide. Aspergillus niger glucose oxidase and Dacytlium dendroides galactose oxidase were used to immunize rabbits and mice in order to trace the distribution of specific antibodies in the immunocytes of the popliteal lymph nodes.
STUDIES IN MYOCLONUS EPILEPSY III. THE EFFECTS OF AMYLOLYTIC ENZYMES ON THE ULTRASTRUCTURE OF LAFORA BODIES
Authors: NIKAIDO TORU , AUSTIN JAMES , STUKENBROK HANS
Published: April 2011

Sections containing Lafora bodies were exposed to the action of amylolytic enzymes. These enzymes split the glucose linkages of glucose polymers (polyglucosans). The following ultrastructural changes were observed: ( a) the fibrils of Lafora bodies were greatly reduced in number; ( b) the staining intensity of the fibrils was reduced; ( c) the amorphous densities were no longer seen. The evidence suggests that the fibrils and amorphous densities of Lafora bodies are both polyglucosan in nature. Lafora bodies resemble corpora amylacea in this respect.
CYTOCHEMICAL MEASUREMENTS OF β-GLUCURONIDASE ACTIVITY IN NORMAL AND INFLAMED HUMAN GINGIVA
Authors: GOGGINS JOHN F. , BILLUPS LLOYD C.
Published: April 2011

Sections of normal and inflamed human gingivae were stained for β-glucuronidase and N-acetylhexosaminidase activities. A marked degree of staining was seen in macrophages and polymorphonuclear leukocytes, whereas fibroblasts, endothelial cells and round cell infiltrates were stained to a lesser extent. In order to quantify the activity in normal and inflamed gingival connective tissues, comparable samples were dissected from adjacent frozen-dried sections of gingival biopsies and assayed for β-glucuronidase activity and deoxyribonucleic acid (DNA) content utilizing microanalytical procedures. Chronically inflamed areas had approximately 4 times the amount of β-glucuronidase activity as normal based on dry weight. However, when the results were expressed in terms of DNA content, there was no statistically significant difference between the normal and inflamed connective tissues. The increase in activity of this enzyme with inflammation in gingiva, therefore, appears to be associated with an increased cellularity rather than caused by an elevation in the average enzyme activity per cell.
THE SIGNIFICANCE OF MUCOPOLYSACCHARIDES IN AMYLOID
Authors: PRAS M. , NEVO Z. , ROTMAN J. , SCHUBERT M. , MATALON R.
Published: April 2011

Amyloid was extracted from six amyloid-laden organs of four different diseases: familial Mediterranean fever (FMF), Hodgkin's disease, rheumatoid arthritis and tuberculosis. In the case of FMF amyloid extracts of three different organs of the same patient were prepared. Chemical studies have indicated the identity of the mucopolysaccharide composition in the amyloid preparations, the amyloid-laden organs and normal organs. The differences were limited to the amounts of the mucopolysaccharide components. The metachromatic properties of an amyloid preparations were not directly related to the level of mucopolysaccharides present. The protein of the amyloid fibers was found to differ chemically from the core protein of chondromucoprotein.
ULTRASTRUCTURAL LOCALIZATION OF CONTRACTILE PROTEIN (THROMBOSTHENIN) IN HUMAN PLATELETS USING AN UNLABELED ANTIBODY-PEROXIDASE STAINING TECHNIQUE
Authors: BOOYSE FRANCOIS M. , ZSCHOCKE DOROTHEA , RAFELSON MAX E. , STERNBERGER LUDWIG A.
Published: April 2011

Soluble horseradish peroxidase-antihorseradish peroxidase (rabbit) complex was used to localize the actomyosin-like contractile protein, thrombosthenin, in both intact human platelets (Epon-embedded) and ultrathin sections (methacrylate-embedded). Antibody staining of ultrathin sections showed the presence of membrane-associated and cytoplasmic thrombosthenin. Antibody staining of intact cells showed that membrane-associated thrombosthenin was localized, at least in part, in the exterior, "fluffy" coat of the platelet. Even after prolonged fixation and/or incubation with the various antisera, we were unable to demonstrate antibody penetration of intact, fixed platelets. Brief treatment with Pronase removed the surface-localized proteinaceous material and completely abolished all antibody staining.
THE CASELLA REACTION AND THE PERMANGANATE-INDUCED BASOPHILIA OF HUMAN ELASTICA RELATED FUNCTIONS OF AROMATIC RESIDUES
Authors: COOPER JOHN H.
Published: April 2011

Casella procedures using dilute neutral permanganate solutions for 5-15 min produce a homogeneous staining of human elastica more suggestive of protein than of carbohydrate determinants. Observed blockage of this reaction by preoxidative acetylation or dinitrophenylation, but not by bromination or nitrosation, indicates tyrosine residues as likely determinants, as does the resemblance of the Casella staining pattern to that yielded by diazotization coupling procedures. By analogy with the known lability of free phenol to dilute neutral permanganate, it is postulated that the alkylphenolic tyrosine residue would thereby be oxidized to alkylcarboxylic acid via an alkylaldehyde intermediate stage. Hence it is significant that prolongation of permanganate treatment leads to progressive diminution of Schiff reactivity with concurrent development of increasing basophilia of carboxylic acid type. In confirmation, the precipitated floccules of a synthetic polytyrosyl peptide (mol wt 60,000) embedded in gelatin were found to give negative direct and periodic acid-Schiff reactions, but a strongly positive Casella reaction and marked permanganate-induced basophilia.
Effect of Therapeutic Modalities on Patients With Patellofemoral Pain Syndrome
Authors: Lake David A. , Wofford Nancy H.
Published: April 2011

Context:

Patellofemoral pain syndrome (PFPS) is a common orthopaedic condition for which operative and nonoperative treatments have been used. Therapeutic modalities have been recommended for the treatment of patients with PFPS—including cold, ultrasound, phonophoresis, iontophoresis, neuromuscular electrical stimulation, electrical stimulation for pain control, electromyographic biofeedback, and laser.

Objective:

To determine the effectiveness of therapeutic modalities for the treatment of patients with PFPS.

Data Sources:

In May and August 2010, Medline was searched using the following databases: PubMed, CINAHL, Web of Science Citation Index, Science Direct, ProQuest Nursing & Allied Health, and Your Journals@OVID.

Study Selection:

Selected studies were randomized controlled trials that used a therapeutic modality to treat patients with PFPS. The review included articles with all outcome measures relevant for the PFPS patient: knee extension and flexion strength (isokinetic and isometric), patellofemoral pain assessment during activities of daily life, functional tests (eg, squats), Kujala patellofemoral score, and electromyographic recording from knee flexors and extensors and quadriceps femoris cross-sectional areas.

Data Extraction:

Authors conducted independent quality appraisals of studies using the PEDro Scale and a system designed for analysis of studies on interventions for patellofemoral pain.

Results:

Twelve studies met criteria: 1 on the effects of cold and ultrasound together, ice alone, iontophoresis, and phonophoresis; 3, neuromuscular electrical stimulation; 4, electromyographic biofeedback; 3, electrical stimulation for control of pain; and 1, laser.

Discussion:

Most studies were of low to moderate quality. Some reported that therapeutic modalities, when combined with other treatments, may be of some benefit for pain management or other symptoms. There was no consistent evidence of any beneficial effect when a therapeutic modality was used alone. Studies did not consistently provide added benefit to conventional physical therapy in the treatment of PFPS.

Conclusions:

None of the therapeutic modalities reviewed has sound scientific justification for the treatment of PFPS when used alone.
Patellofemoral Disorders
Authors: Parikh Shital , Noyes Frank R.
Published: April 2011

Background:

The contribution of lower limb rotational malalignment to patellofemoral pain and instability has been well recognized. The purpose of the present study is to review the role of computed tomography (CT) and magnetic resonance imaging (MRI) in assessment of abnormal rotational alignment of lower limb.

Evidence Acquisition:

An analysis of all available literature in the English language through 2010 was performed to provide data on a comparison between MRI and CT—specifically, the techniques and normative values used to determine abnormal lower limb alignment.

Results:

CT and MRI are highly accurate in defining abnormal alignment of the lower limb. Determination of axis of femoral anteversion in proximal femur has been the subject of debate in the literature. The determination of distal femoral condylar axis, proximal tibial axis and distal tibial axis are less controversial.

Conclusions:

CT and MRI are both used for assessing the rotational abnormalities of the femur and tibia during evaluation for patellofemoral disorders. MRI has an advantage over CT because femoral anteversion measurements are more accurate and ionizing radiation is avoided. A standardized protocol defining the level and axes for measurement of femoral and tibial alignment indices should be used to maintain consistency in measurements.
Prevalence of Hyponatremia, Renal Dysfunction, and Other Electrolyte Abnormalities Among Runners Before and After Completing a Marathon or Half Marathon
Authors: Mohseni Michael , Silvers Scott , McNeil Rebecca , Diehl Nancy , Vadeboncoeur Tyler , Taylor Walt , Shapiro Shane , Roth Jennifer , Mahoney Sherry
Published: April 2011

Background:

Prior reports on metabolic derangements observed in distance running frequently have small sample sizes, lack prerace laboratory measures, and report sodium as the sole measure.

Hypothesis:

Metabolic abnormalities—hyponatremia, hypokalemia, renal dysfunction, hemoconcentration—are frequent after completing a full or half marathon. Clinically significant changes occur in these laboratory values after race completion.

Study Design:

Observational, cross-sectional study.

Methods:

Consenting marathon and half marathon racers completed a survey as well as finger stick blood sampling on race day of the National Marathon to Fight Breast Cancer (Jacksonville, Florida, February 2008). Parallel blood measures were obtained before and after race completion (prerace, n = 161; postrace, n = 195).

Results:

The prevalence of prerace and postrace hyponatremia was 8 of 161 (5.0%) and 16 of 195 (8.2%), respectively. Hypokalemia was not present prerace but was present in 1 runner postrace (1 of 195). Renal dysfunction occurred prerace in 14 of 161 (8.7%) and postrace in 83 of 195 (42.6%). Among those with postrace renal dysfunction, 45.8% (38 of 83) were classified as moderate or severe. Hemoconcentration was present in 2 of 161 (1.2%) prerace and 6 of 195 (3.1%) postrace. The mean changes in laboratory values were (postrace minus prerace): sodium, 1.6 mmol/L; potassium, −0.2 mmol/L; blood urea nitrogen, 2.8 mg/dL; creatinine, 0.2 mg/dL; and hemoglobin, 0.3 g/dL for 149 pairs (except blood urea nitrogen, n = 147 pairs). Changes were significant for all comparisons ( P < 0.01) except potassium ( P = 0.08) and hemoglobin ( P = 0.01).

Conclusions:

Metabolic abnormalities are common among endurance racers, and they may be present prerace, including hyponatremia. The clinical significance of these findings is unknown.

Clinical relevance:

It is unclear which runners are at risk for developing clinically important metabolic derangements. Participating in prolonged endurance exercise appears to be safe in the majority of racers.
METACHROMATIC BASOPHILIA OF KERATIN AFTER OXIDATION-CLEAVAGE OF DISULFIDE BONDS
Authors: LILLIE RALPH D. , BANGLE RAYMOND , FISHER EDWIN R.
Published: April 2011

Oxidation of keratinous structures by peracetic acid, bromine in carbon tetrachloride, or neutral potassium permanganate produced an insoluble, strongly acid substance capable of taking up basic dyes even from pH 1.2 solution. Thus, hair shafts, nails, and, to a much less extent, stratum corneum of the epidermis were rendered basophilic and metachromatic. The oxidation product strongly resisted methylation, thereby differing from the basophil substance of cell nuclei, of mast cell granules, and of cartilage. Furthermore, the oxidation product was not blocked by aniline chloride or phenylhydrazine, indicating that aldehydes, if formed, were not responsible for the basophilia by the formation of Schiff's bases.

Periodic acid (1% aqueous solution for 2 hours at 60°C.) also rendered keratinous structures basophilic but to a less intensity than with the other oxidizing agents. Chromic acid was ineffective under the conditions used and, moreover, largely destroyed nuclear basophilia.

The various oxidizing agents that induced metachromatic basophilia in keratin have in common the property of cleaving the disulfide bonds of cystine, yielding cysteic acid. In the case of oxidized keratin, it appears that the resultant cysteic acid residues remain bound in peptide chains but still possess free sulfonic acid radicals. Since sulfonic acids are strong acids, such groups probably account for the observed metachromatic basophilia in oxidized keratinous structures.
STUDIES IN HISTOCHEMISTRY XXX. A STUDY BY PHASE CONTRAST MICROSCOPY OF CYTOLOGICAL EFFECTS OF FREEZING-DRYING PROCEDURES ON CULTURED FIBROBRASTS AND GUINEA PIG TISSUES
Authors: BLOOM DAVID , SWIGART RICHARD H. , SCHERER WILLIAM F. , GLICK DAVID
Published: April 2011

Observations made by phase contrast microscopy have been reported for the effects of formalin fixation, freezing-thawing, and freezing-drying on tissue culture fibroblasts. The fibroblasts appeared somewhat shrunken after formalin fixation, and the cellular structures appeared to be more clearly delineated. A granular cytoplasm and "brilliant" nuclei were observed. Freezing-thawing resulted in minimal change, but paraffin embedding and deparaffinization had a marked effect on the intracellular morphology. Similar studies were made on frozen-dried tissues with and without paraffin infiltration. Fresh frozen-dried sections prepared at –40° in a cryostat had structures which appeared to be more distinct than those from sections of paraffin embedded tissues prepared at –40° in the Glick-Malmstrom apparatus; however, the former frequently contained distorted areas.
STUDIES IN HISTOCHEMISTRY XXXI. A METHOD FOR THE DETERMINATION OF PROTEIN IN MILLIMICROGRAM QUANTITIES
Authors: NAYYAR SOM N. , GLICK DAVID
Published: April 2011

The spectrophotometric determination of protein in millimicrogram quantities was described which is suitable for analysis of microtome sections of tissue and other samples of histochemical interest. The macro procedure of Greif was adapted to the micro scale and a modification was employed in which protein was quantitatively precipitated with an excess of bromsulfalein at a low pH and the excess dye was photometrically determined after addition of alkali. Kjeldahl nitrogen analysis of the protein was used for standardization and the results were expressed in terms of protein-nitrogen. With samples and equipment described, a standard error of 1.4% corresponding to 0.9 mµg. of protein-nitrogen was observed. With smaller pipettes and the same cuvettes the volumes employed could be rather easily reduced to about one half with a doubling of the sensitivity of the method. Further increase in sensitivity could be achieved without inconvenience by the use of cuvettes of smaller diameter, e.g. 0.7-0.8 mm., and pipettes of correspondingly reduced capacities.
THE DEGRADATION BY RIBONUCLEASE OF SUBSTRATES OTHER THAN RIBONUCLEIC ACID
Authors: MCDONALD MARGARET R. , KAUFMANN BERWIND P.
Published: April 2011

These experiments support the premise which is the basis for cytochemical procedures that employ ribonuclease for the identification of ribonucleic acid, namely, that the substrate specifically degraded by ribonuclease is ribonucleic acid. The reduction in Feulgen stainability of chromatin effected by ribonuclease has been shown to be due to an intracellular deoxyribonuclease, which hydrolyzes intracellular deoxyribonucleic acid only after ribonucleic acid has been removed. The degradation of apurinic acid by ribonuclease can be duplicated by other proteins.
RIBONUCLEASE AND RIBONUCLEIC ACID SYNTHESIS
Authors: HOKIN LOWELL E. , HOKIN MABEL R.
Published: April 2011

The specific activities of the ribonucleotides in various fractions of complete ribonuclease digests of P32-labelled RNA have been compared. The specific activity of either of the purine nucleotides is essentially the same in the dialysate and in the core. The specific activities of the pyrimidine nucleotides of the dialysate are higher than the specific activities of the corresponding nucleotides in the core. The specific activities of the pyrimidine mononucleotides are higher than those of the corresponding pyrimidine nucleotides bound as polynucleotides; this can account for the higher specific activities of the pyrimidine nucleotides in the dialysate as compared to the core. The specific activities of the pyrimidine nucleotides bound in polynucleotides appear to be the same in core and dialysate. These data show that pyrimidine nucleotides which are esterified through the 5' hydroxyl of their ribose to the secondary phosphoryl group of an adjacent pyrimidine 3' nucleotide are metabolically more labile than pyrimidine nucleotides esterified in a similar manner to adjacent purine 3' nucleotides. Since ribonuclease attacks the former linkage, but not the latter, the data presented here suggest that ribonuclease may play a role in the turnover of RNA.
FASTER HISTOCHEMICAL REACTION FOR ATPASE IN PRESENCE OF CHLORIDE SALTS, WITH STUDIES OF THE MECHANISM OF PRECIPITATION
Authors: BERG GEORGE G. , LYON DOROTHEA , CAMPBELL MARILYN
Published: April 2011

A very rapid localization of nucleoside triphosphate phosphohydrolase activity is given by incubation mixtures containing 2 M chlorides of monovalent cations. An optimal procedure is to fix overnight in cold formaldehyde-calcium, store in gum-sucrose, freeze-section and incubate 4 min or less at 30°C in a solution of 3 mM Na2 adenosine triphosphate (ATP), 2 M KCl, 3 mM MgCl2, and 1 mM Pb(NO3)2 in 24 mM Tris buffer adjusted to pH 7.2 with HCl. In rat intestine, for example, a strong, selective stain was developed in myenteric plexus, blood vessels, lamina propria mucosae and smooth muscle, while paired preparations without 2 M KCl were pale or unstained. The reaction was specific to nucleoside triphosphates and required Mg++. Acceleration by 2 M chloride was also found in the adenosine triphosphate phosphohydrolase reaction given by a single protein fraction (thin disc) in acrylamide gel electropherograms. The histochemical reaction for ATP phosphohydrolase produced residues of lead in formaldehyde-fixed sections, with two-thirds of lead captured as a precipitate of products of enzyme reaction and one-third bound directly to tissue. Only the enzyme-generated precipitate gave an anatomically and chemically selective and time-dependent pattern of staining, while direct binding gave a diffuse and time-independent background stain enhanced by omitting the substrate. Less than 1% of precipitate originated from nonenzymatic breakdown of ATP. Conditions of precipitation were studied. Reaction products were captured by lead as mixed crystals of nucleotides and phosphate. Addition of chloride salts decreased the concentration of lead required to saturate the reaction mixture, and this effect probably accounted for the enhanced histochemical reaction.
ANALYSIS OF FLUORESCENCE EXCITATION PEAK RATIOS FOR THE CELLULAR IDENTIFICATION OF NORADRENALINE, DOPAMINE OR THEIR MIXTURES
Authors: BJÖRKLUND ANDERS , EHINGER BERNDT , FALCK BENGT
Published: April 2011

Treatment with HCl induces changes in the excitation spectra of the formaldehyde condensation products of dopamine and noradrenaline that permit the distinction of these two catecholamines on the cellular level. Using histochemical models and tissue sections, these spectral shifts and changes in the ratio of the two main excitation peaks at about 320 and 370 nm have been measured. Acidification of the dopamine fluorophore rapidly leads to the conversion of the quinoidal dihydroisoquinoline fluorophore into its tautomeric nonquinoidal form. The process probably goes to completion within a few seconds following treatment with HCl. The fluorophore remains stable during prolonged acid treatment. The conversion of the noradrenaline fluorophore into its nonquinoidal state is equally rapid; however, the subsequent dehydration to the fully aromatic form is a slower process, and it is apparently not complete even after several minutes, when the rising over-all tissue fluorescence makes measurement increasingly difficult. The results, however, indicate that this characteristic fluorophore is formed from the noradrenaline fluorophore in a high degree also in tissue sections. In the present report, the various states of the catecholamine fluorophores are characterized in more detail, and the evaluation of excitation spectra for the cellular identification of dopamine and noradrenaline, alone or in mixtures, is discussed. A detailed description of the practical performance of the microspectrofluorometric differentiation procedure is given.
MICROBODIES IN LEYDIG CELL TUMORS OF RAT TESTIS
Authors: REDDY JANARDAN , SVOBODA DONALD
Published: April 2011

Spontaneous Leydig cell tumors (interstitial cell tumors) of the testis arising in aged rats have been studied by cytochemical procedures for identification of microbodies. Large numbers of microbodies with diameters ranging from 0.1 to 0.4 µ were present in tumor cells. The size of the tumor did not appear to have any effect on the microbody number in neoplastic Leydig cells. Examination of large numbers of microbodies in tumor cells revealed continuities between adjacent microbodies. The microbodies in Leydig cell tumors lacked a typical crystalline core or nucleoid, but some of the elongated forms contained cylindrical or tubular structures with periodicity. The nature of these inclusions remains obscure because urate oxidase activity was not detected in these tumors, by either biochemical or histochemical methods. In view of the presence of numerous microbodies, it is suggested that these tumors serve as a rich source of microbodies for studying their functional significance in Leydig cells of the testis.
THE IN VITRO INTERCONVERSION BY MERCAPTANS OF TYPE I GLUCOSE 6-PHOSPHATE DEHYDROGENASE ISOZYMES OF RAT
Authors: HORI SAMUEL H. , YONEZAWA SATOSHI
Published: April 2011

The rat glucose 6-phosphate dehydrogenase isozymes of cell sap origin designated D, F1, F2 and F3 were found to be interconvertible with each other in the presence or absence of mercaptans. The D and F3 enzymes were the oxidized, the F1 was the reduced and the F2 was the intermediate form. Molecular weight estimation by the polyacrylamide gel electrophoresis and Sephadex G-200 methods demonstrated that the D enzyme had twice the molecular weight of the F enzymes. The mitochondrial enzyme which moved as fast as the F3 enzyme of cell sap origin was not modified with mercaptans. Together with previous data, the present results caution in the interpretation of zymograms.
DEMONSTRATION OF HISTONES IN PROERYTHROBLASTS IN PERNICIOUS ANEMIA AND THE DI GUGLIELMO SYNDROME
Authors: KASS LAWRENCE
Published: April 2011

Cytochemical studies on histones of proerythroblasts from patients with pernicious anemia and the Di Guglielmo syndrome were performed. Reduced amounts of histones were found in the proerythroblasts of pernicious anemia using the alkaline fast green stain. Ammoniacal silver staining indicated predominantly lysine-rich histone in the nuclei of pernicious anemia proerythroblasts. Using the same stain, the nuclei of Di Guglielmo proerythroblasts demonstrated a "speckled" pattern, indicating discrete deposits of arginine-rich histone. The relationships which may exist between these findings and the pathogenesis of megaloblastic and megaloblastoid erythropoiesis are discussed.
THE RELATION OF THE PROPERTIES OF CONGO RED-STAINED AMYLOID FIBRILS TO THE β-CONFORMATION
Authors: GLENNER G. G. , PAGE D. L. , EANES E. D.
Published: April 2011

Amyloid fibrils and the aminoterminal variable fragment of some Bence Jones proteins are known to have Congo red affinity and, after staining, to exhibit dichroism, polarization birefringence and a green polarization color. X-ray crystallographic analysis demonstrates the presence in these preparations of a 4.72-4.76Å d-spacing indicating an antiparallel chain β-pleated sheet structure as the major conformation. Only the β-form of poly- l-lysine has all of these properties. It is suggested that the tinctorial and polarization optical properties of amyloid deposits stained with Congo red may be at least in part dependent upon the presence of the β-structure as the major protein conformation of amyloid fibrils.
DISTRIBUTION OF MUCOSUBSTANCES IN THE DEVELOPING RAT HEART
Authors: MARKWALD ROGER R. , SMITH WILLIAM N. ADAMS
Published: April 2011

Mucosubstances (MS) were examined in 10½-14½-day embryonic rat hearts utilizing nonaqueous fixatives or formaldehyde vapor-fixed frozen sections hydrated in concentrated solutions of cetylpyridinium chloride. Ribonuclease-resistant, polyanionic sites were limited to the extracellular cardiac jelly, endocardium and fibroblastic cells (cushion tissue) associated with the endocardium. The cardiac jelly and endocardium of day 10½ embryos principally contained a hyaluronic acid-like carboxylated mucosubstance whose alcianophilia at pH 2.5 was abolished by hyaluronidase but was resistant to NaOH extraction and neuraminidase and trypsin digestion. A critical electrolyte concentration of 0.2 M MgCl2 abolished alcianophilia. On days 13½-14½ carboxylated MS were restricted to cushion tissue and partially resisted mild methylation. Sulfated MS were limited to primitive endocardial cells which gave origin to cushion tissue. Dye deposits of aldehyde fuchsin, high iron diamine or Alcian Blue (pH 1.0) were localized on cell surfaces and such staining was prevented by strong (60°C) methylation. Hyaluronidase sensitivity of sulfated MS decreased with gestation. The critical electrolyte concentration varied from 0.5-0.7 M MgCl2 on days 11½-12½ to 0.8-0.9 M MgCl2 after day 12½. The sulfated MS of endocardial cells were preceded by a transitory accumulation of diastase-resistant, periodic acid-Schiff-positive material. Possible roles of MS in normal and abnormal cardiac septation processes are discussed.