bluuebunny/crossref_metadata_2025 huggingface dataset:
LOCALIZATION OF NICOTINAMIDE ADENINE DINUCLEOTIDE PHOSPHATE-DEPENDENT DEHYDROGENASES IN CATECHOLAMINE-CONTAINING NEURONS OF RAT BRAIN STUDIES ON THE NUCLEUS LOCUS CERULEUS
Authors: KAUFFMAN F. C. , BLOOM F. E. , SIMS K. L. , PICKEL V. M.
Published: April 2011

Histochemical and cytochemical methods have been used to localize glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, isocitrate (nicotinamide adenine dinucleotide phosphate (NADP)) dehydrogenase and malic enzyme in the nucleus locus ceruleus and other catecholamine-containing neuronal groups of the rat brain stem. The four NADP-dependent dehydrogenases were studied by both quantitative and qualitative histochemical techniques using adjacent tissue sections. Both types of analyses were done on neuronal nuclei known to contain catecholamines in high concentration, the nucleus locus ceruleus and the superior cervical sympathetic ganglion; other known catecholamine-containing nuclei were surveyed by the cytologic technique only. There was intense staining of cell bodies and neuropil of the nucleus locus ceruleus after all four of the NADP dehydrogenase histochemical reactions. In contrast, little staining was observed in the adjacent vestibular nuclei or mesencephalic root nucleus of the trigeminal with the exception of appreciable glucose 6-phosphate dehydrogenase activity present in neuropil elements. Quantitative microchemical determinations in the nucleus locus ceruleus corroborate the histochemical results which indicated high NADP dehydrogenase activities particularly for 6-phosphogluconate dehydrogenase. This same pattern of high NADP enzyme activity as determined by combined cytochemical and quantitative chemical techniques was also observed in the superior cervical ganglion and cytochemically in other catecholamine-containing nuclei of the brain stem. Our findings suggest that a high capacity to generate or utilize nicotinamide adenine dinucleotide phosphate (reduced) may be characteristic of those neurons which either receive adrenergic terminals or synthesize catecholamines.
DIETHYLENE GLYCOL DISTEARATE AS AN EMBEDDING MEDIUM FOR HIGH RESOLUTION LIGHT MICROSCOPY
Authors: TALEPOROS PLATO
Published: April 2011

Owing to its physical properties, diethylene glycol distearate is especially suited for obtaining thin sections of high quality with little or no compression. Tissues can be cut as thin as 1 µm on a steel knife without cooling, but a water trough is essential. The wax is translucent, so that specimens can be oriented for a desired plane of sectioning. The wax is also of sufficient hardness to serve as an embedding medium for osmium-fixed material and can readily be removed from such tissue sections for rapid phase contrast studies. A general method is described for embedding and sectioning with diethylene glycol distearate wax and examples of freeze-substituted and osmium-fixed material are illustrated. It is concluded that this synthetic wax can offer a convenient and simple means for obtaining thin sections for most aspects of high resolution light microscopy.
PREPARATION AND USE OF SOLUBLE FERRITIN-ANTIFERRITIN COMPLEXES AS A SPECIFIC MARKER FOR IMMUNOELECTRON MICROSCOPY
Authors: MARUCCI AMERICO A. , DOUGHERTY ROBERT M. , DISTEFANO HENRY S.
Published: April 2011

Soluble immune complexes of ferritin and chicken antiferritin have been prepared. This has been used with unlabeled chicken antibody to avian leukosis virus and rabbit antichicken globulin to demonstrate the presence of specific viral antigens in tissue culture by electron microscopy.
CYTOCHEMICAL DEMONSTRATION OF HISTONASE ACTIVITY IN HUMAN LEUKOCYTES
Authors: KASS LAWRENCE , SCHNITZER BERTRAM
Published: April 2011

A cytochemical technique was devised for demonstrating a lysosomal enzyme capable of hydrolyzing histone. The test for histonase involved incubation of peripheral blood leukocytes in a mixture of calf thymus histone (substrate) and Fe(NH4)2(SO4)2·6H2O. Combination of this iron salt occurred with products of histone cleavage (arginine, lysine, glycine). The iron-amino acid complex was made visible in the cytoplasm of the cell by the Prussian blue stain. Histonase enzyme was found to have a narrow substrate specificity and to react only with histone and not with a variety of biologic and synthetic substrates for protease activity. The role of histonase in the transfer of information from nucleus to cytoplasm is discussed.
NUCLEOSIDE DIPHOSPHATASE IN THE ONION ROOT TIP I. EFFECTS OF FIXATION AND LEAD ON ENZYME ACTIVITY
Authors: GOFF C. W. , KLOHS W. D.
Published: April 2011

The terminal 0.2—0.5 mm of 3-day onion roots grown from bulbs were excised and fixed for 1½ hr in various concentrations of cacodylate-buffered glutaraldehyde, formaldehyde or combinations of these. Alternatively, the fixative concentration was held constant while fixation time was varied. Control roots were run in buffer lacking fixative. The roots were then homogenized and in most cases aliquots of the entire homogenate were used to assay for total nucleoside diphosphatase (NDPase). Parallel assays were usually run after treating the homogenate with deoxycholate. Both glutaraldehyde and formaldehyde inhibited NDPase activity, the extent of this inhibition depending upon fixation time, fixative concentration and the particular aldehyde used, although with both fixatives inhibition did not increase beyond a certain level even with further increase in fixation time or fixative concentration. With glutaraldehyde and glutaraldehyde-containing fixatives, this level was normally about 74% inhibition while with formaldehyde it was about the level of non-activated NDPase activity and inhibition could be detected only after deoxycholate treatment. Lead did not appear to inhibit the "fixed" NDPase.
THE HISTOCHEMICAL INTERPRETATION OF THE COMPLEX RESULTS OF METHYLATION UPON GASTROINTESTINAL TRACT MUCINS, WITH SPECIAL REFERENCE TO THE PERIODIC ACID-SCHIFF REACTIVITY
Authors: REID P. E. , CULLING C. F. A. , DUNN W. L.
Published: April 2011

The histochemical use of methylation has complex results; particularly in respect of the periodic acid-Schiff reaction, these are analyzed and discussed. Methods are described which allow the separate study of the following effects: (a) the removal of the KOH/periodic acid-Schiff effect; (b) removal of sialic acid from a potential vicinal diol; and (c) the removal of O-sulfate ester from a potential vicinal diol. The use of the Smith degradation technique, in addition to the above, also allows inferences to be drawn in respect of the structure of the mucins (glycoproteins) being investigated.
ELECTRON MICROSCOPIC LOCALIZATION OF PHYTOCHROME IN PLANTS USING AN INDIRECT ANTIBODY-LABELING METHOD
Authors: COLEMAN RICHARD A. , PRATT LEE H.
Published: April 2011

The use of polyethylene glycol as a preembedding medium for immunocytochemistry at the electron microscope level has been adapted to the localization of phytochrome in etiolated oat (Avena sativa L., cv. Garry) seedlings. Phytochrome was indirectly labeled in 3-µm sections with rabbit antiperoxidase-peroxidase complex using sheep antirabbit serum and a specific rabbit antiphytochrome serum. Following localization the 3-µm sections were reembedded for ultrathin sectioning. In the absence of information regarding the subcellular distribution of phytochrome, it was necessary to develop a control which would demonstrate that all organelles and areas of the tissue being localized were penetrated by all reagents. Such a control is described. In those cells which contained phytochrome it was found to be generally distributed throughout the cytoplasm and to be associated with both amyloplasts and mitochondria. No activity was observed in nuclei.
PEROXIDASE-LABELED ANTIBODY A NEW METHOD OF CONJUGATION
Authors: NAKANE PAUL K. , KAWAOI AKIRA
Published: April 2011

A new method of conjugating horseradish peroxidase with proteins was developed. The carbohydrate moiety of fluorodinitrobenzene-blocked peroxidase was oxidized with sodium periodate to form aldehyde groups. The peroxidase-aldehyde was then bound to free amino groups of proteins unidirectionally at high efficiencies. Peroxidase-labeled immunoglobulin retained its immunologic as well as enzymatic activities.
DETERMINATION OF THE POLARIZATION OPTICAL PROPERTIES OF THE AMYLOID-CONGO RED COMPLEX BY PHASE MODULATION MICROSPECTROPHOTOMETRY
Authors: TAYLOR DOUGLASS L. , ALLEN ROBERT D. , BENDITT EARL P.
Published: April 2011

The polarization properties responsible for the classical "green birefringence" of the amyloid-Congo red complex have been determined by a new optical method, phase modulation microspectrophotometry. This method now makes possible the measurement of one optical property at a time (birefringence, optical rotation, linear dichroism and circular dichroism throughout the visible spectrum) in complex specimens in which visible contrast in polarized light is the result of a mixture of polarization effects. The green birefringence is explained by a combination of optical effects, the strongest of which are dispersion of birefringence and linear dichroism superimposed on the smaller effects of circular dichroism and optical rotatory dispersion. The interaction of the planar dye molecules with the amyloid protein induces an extrinsic Cotton effect.
QUANTITATIVE COMPARISONS OF AMINE FLUORESCENCE IN CORTICAL NORADRENALINE TERMINALS USING SMEAR PREPARATIONS
Authors: AGNATI LUIGI F. , FUXE KJELL
Published: April 2011

A method is introduced by which quantitative estimations of changes in amine fluorescence of cortical noradrenaline (NA) terminals can be made using smear preparations (Olson and Ungerstedt, 1970). The method is based on counts performed on photographs using different transmittance gratings. With use of this method it is possible to make comparisons based not only on the number of NA varicosities, but also on the fluorescence intensity of the sampled fields. The method is highly reliable, since the data are in good agreement with biochemical findings as illustrated in experiments using tyrosine-hydroxylase inhibition and 6-hydroxydopamine-induced lesions of the NA pathway to the cortex cerebri.
A PROCEDURE FOR THE USE OF ATOMIC ABSORPTION SPECTROPHOTOMETRY IN QUANTITATIVE HISTOCHEMISTRY
Authors: ROSENQUIST THOMAS H. , ROSENQUIST JUDITH W.
Published: April 2011

In this quantitative histochemical study, tissues were sectioned, then stained with procedures that conferred a metallic precipitate. Sections were photographed, removed from the slides, air-dried and weighed and then dissolved in acid and the metal content analyzed by atomic absorption spectrophotometry. The stains used in this study were the Alcian Blue (copper phthalocyanin) critical electrolyte concentration procedure on sections of the heart and duodenum from young male rats, colloidal iron on sections of the kidney from adolescent and senescent rats, silver impregnation on sections of the kidney of adolescent and senescent rats and an adenosine triphosphatase lead precipitation reaction on sections of the liver, kidney and heart from adolescent and senescent mice. The objective data obtained by the use of atomic absorption spectrophotometry, reported as micrograms of metal per milligram of dry weight of tissue, allowed statistical analysis of any differences in stain intensity of metallic dyes or metal-precipitating histochemical reactions. Attributes of atomic absorption spectrophotometry which make it desirable for use in this application include accuracy, ease of operation, low cost and ready availability.
EVALUATION OF CORRELATIONAL INFORMATION IN DIGITIZED CELL IMAGES
Authors: JETER WAYBURN S. , OLSON GEORGE B. , BARTELS PETER H. , BAHR GUNTER F. , TAYLOR JOHN , WIED GEORGE L.
Published: April 2011

The processing of digitized cell images by computer may yield information not only about a set of cell image properties but also about their mutual dependencies. Observation of the covariance matrix of image properties of cellular material showing a response to chemotherapeutic treatment, ionizing radiation or antigenic challenge or following a developmental trend may permit a quantitative description of small trendal charges The covariance between certain cell image properties may show statistically significant changes before the mean values of the image properties are affected. Methods of reducing the dimensionality of the representation in an efficient manner are described.
SENSITIVITY IN ELECTRON MICROSCOPE AUTORADIOGRAPHY FOR 125I
Authors: FERTUCK HELEN C. , SALPETER MIRIAM M.
Published: April 2011

Sensitivity in electron microscope autoradiography was determined for 125I. Values are given using Ilford L4 and Kodak NTE emulsions combined with different developers. The extent of self-absorption as a function of section thickness and heavy metal staining and the effect of radiation dose (dose dependence) were assessed. It was found that the over-all efficiency for 125I was better than that for tritium and that, as is the case with tritium, there is a distinct "dose dependence" especially when Microdol X is the developer. Self-absorption studies indicate that self-absorption is increased by about 15% when the specimen thickness is increased from 300 to 1000 Å. An increase of under 15% is also introduced by heavy metal staining of sections in this thickness range.
OBSERVATIONS IMPLICATING AN EXTRACELLULAR ENZYMIC MECHANISM OF CONTROL OF BONE MORPHOGENESIS
Authors: URIST MARSHALL R. , IWATA HISASHI , BOYD STUART D. , CECCOTTI PETER L. , OKADA MARLYS , UYENO SATSUKI , KIRKPATRICK SUE , KING EDWARD
Published: April 2011

Data on physicochemical conditions leading to loss of the bone morphogenetic property of bone matrix in neutral buffer solutions support the concept of an enzymic control mechanism better than a chemical blocking reaction or denaturation. The loss is associated with release of 35S-labeled constituents and not prevented by ε-amino caproic acid, an inhibitor of cathepsins. The loss is also associated with release of 35S-cysteine-labeled protein; about 60% of the yield is sustained by the addition of only 3 mmoles/liter of iodoacetic acid. A latent period of about 12 hr, decreased by extraction of bone matrix with CaCl2, is characterized by release of protein polysaccharide and other noncollagenous proteins. Release of sialic acid from the bone matrix by neuraminidase at pH 7.4 has no effect upon bone yield. At 2°C, Tris-HCl buffer or ethylenediaminetetraacetic acid extracts noncollagenous proteins without loss of bone yield; at 37°C, pH 7.4, these solutions also activate endogenous enzymes and reduce bone yield. The component of bone matrix responsible for reduction in bone yield is separable from bone matrix by extraction with phosphate buffer, by catheptic digestion of bone matrix in acidic buffer solutions, by sequential chemical extraction of noncollagenous proteins with cold slightly acidic salt solutions or by extraction-denaturation with chloroform-methanol. Detergents neither extinguish nor denature the morphogenetic property but some solubilize or extract degradative enzymes; hexodecyl trimethyl ammonium bromide, at pH 5.0, is positively charged and extracts hydrophobic proteins, including part of the bone morphogenetic property. A special selection of sulfhydryl chemical inhibitors remarkably different from the selection inhibiting known enzymes preserves the bone morphogenetic property of bone matrix; p-chloromercuribenzoate preservation is reversible by chemical reactions with cysteine. Reduction in bone yield in phosphate buffer is not attributable to a chemical block because chloroform-methanol extraction of the agent does not restore bone yield and is not attributable to denaturation because bone yield sustained by p-chloromercuribenzoate is lost by chemical reactions with cysteine. An hypothetical insoluble bone morphogenetic protein (BMP) firmly bound to collagen is degraded by a soluble neutral proteinase (BMPase). Digestion of the hypothetical BMP occurs without loss of the 640-A electron micrographic image of bone collagen, resembles tryptic digestion and is more selective as well as physiologic in action.
ELECTRON PROBE MICROANALYSIS OF SKELETAL MUSCLE
Authors: YAROM R. , CHANDLER J. A.
Published: April 2011

X-ray microanalysis using the analytical electron microscope (EMMA 4) was performed on resting frog sartorius muscles fixed by the pyroantimonate technique. Calcium was the main element of interest analyzed. Pretreatment was with ethyleneglycoltetraacetate to chelate calcium and with Tris chloride to replace sodium. Osmium-fixed controls were also analyzed. Both crystal-diffracting and energy-dispersive analyzers were employed, the former for accurate quantitation of calcium content and the latter for qualitative assessment of other elements present. Significant concentrations of calcium occurred in the dense parts of the nucleus, the triads, the thin filaments and the sarcolemma. The amount of calcium was greatly decreased by ethyleneglycoltetraacetate pretreatment and slightly increased by sodium removal. After osmium fixation alone, little calcium was detected with no localization.
SEX AND STOCK DIFFERENCES IN THE HISTOCHEMICAL MYOFIBRILLAR ADENOSINE TRIPHOSPHATASE REACTION IN THE SOLEUS MUSCLE OF THE MOUSE
Authors: VAUGHAN HILARY S. , AZIZ-ULLAH . , GOLDSPINK G. , NOWELL N. W.
Published: April 2011

Histochemical myofibrillar adenosine triphosphatase (ATPase) reaction has been studied in the soleus muscles of males, castrate males and females of two different stocks of mice. Males were shown to have a significantly lower percentage of fibers with an alkali-labile ATPase in the soleus than either castrate males or females. The castrate males and females did not differ significantly from each other. This suggests that androgens can influence fiber type ratios. The two stocks of mice differ significantly with respect to the proportion of fibers with an alkali-labile ATPase in the soleus muscle. This is interpreted as a manifestation of genetic differences between the stocks.
CYTOCHEMICAL DETECTION OF HYDROLASES IN FUNGUS CELLS III. ARYL SULFATASE
Authors: REISS JÜRGEN
Published: April 2011

In the cells of Aspergillus oryzae, Aspergillus nidulans, Neurospora crassa and Saccharomyces cerevisiae, aryl sulfatase can be demonstrated by incubation in a medium containing 6-bromo-2-naphthylsulfate as substrate and fast garnet GBC as coupling agent. Controls confirm the specificity of the reaction. Other incubation solutions (two Gomori media and a simultaneous coupling procedure with 8-hydroxyquinoline sulfate as substrate) gave negative results or reaction pictures equally to those in substrate-free control. The possible reasons for this are discussed. In the mycelial fungi the strongest enzyme activity is located in the most intensely metabolizing parts: tips of the hyphae and the differentiating parts of the conidiophores. The reaction granules in all four fungi are possibly identical with lysosomes.
GIANT LYSOSOMES IN CULTURED MONKEY KIDNEY CELLS
Authors: VLADUTIU GEORGIRENE D. , ROSE NOEL R.
Published: April 2011

Following staining with an esterase-specific rabbit antiserum, using the indirect immunofluorescent technique, a large body was observed in close apposition to the nucleus of cultured monkey kidney cells. It is present mainly during the lag and early logarithmic phases of growth. The available evidence suggests that the structure is an autophagic giant lysosome.
TISSUE FIXATION WITH DIIMIDOESTERS AS AN ALTERNATIVE TO ALDEHYDES I. COMPARISON OF CROSS-LINKING AND ULTRASTRUCTURE OBTAINED WITH DIMETHYLSUBERIMIDATE AND GLUTARALDEHYDE
Authors: HASSELL JOHN , HAND ARTHUR R.
Published: April 2011

Diimidoesters are bifunctional reagents which react with the ε-amino group of lysine, forming intermolecular cross-links with minimal alteration of the native properties of proteins. To determine the optimal conditions for tissue fixation with diimidoesters, a cross-linking assay was developed based on the assumption that protein aggregates formed by intermolecular cross-links would be water-insoluble. With this assay method, the proportion of insoluble protein in fresh liver was 16%, while optimal conditions for cross-linking with the diimidoester, dimethylsuberimidate (DMS) (16-20 mg/ml, 0.02 M Ca++ and 0.15 M Tris-HCl, pH 9.5), rendered 92.1% insoluble and permitted only 3.3% of the protein to diffuse into the fixative solution. The smaller diimidoesters, dimethyladipimate and dimethylmalonimidate, rendered 74 and 16% of the protein insoluble, respectively. Fixation with various aldehydes produced insoluble fractions varying from 74% to over 90%. Ultrastructurally, hepatocytes of DMS-fixed liver and glutaraldehyde-fixed liver were similar except that with the former fixative the Golgi saccules and smooth endoplasmic reticulum were dilated and mitochondrial matrices exhibited an increased electron density. Furthermore, the appearance of glutaraldehyde- and DMS-fixed liver was more readily correlated with the degree of cross-linking than with the pH of the fixative solution per se. The results of this study indicate that DMS is a potentially useful fixative for light and electron microscopy.
ULTRAFINE STRUCTURE OF GLYCOGEN PARTICLES SYNTHESIZED UNDER HISTOCHEMICAL CONDITIONS
Authors: TAKEUCHI TADAO
Published: April 2011

The polyglucose particles histochemically synthesized by enzyme activities, phosphorylase and branching glycosyltransferase were clearly demonstrated as a macromolecular spheroidal branching body suggesting the chemical structure proposed by French (2), although the synthesized polyglucose was strikingly larger.
LOCALIZATION WITHIN CLONED RAT PITUITARY TUMOR CELLS OF MATERIAL THAT BINDS ANTIGROWTH HORMONE ANTIBODY
Authors: MASUR SANDRA KAZAHN , HOLTZMAN ERIC , BANCROFT F. CARTER
Published: April 2011

Cultured rat pituitary cells known to synthesize growth hormone were fixed and exposed to antigrowth hormone antibody conjugated to horseradish peroxidase. The cells were then incubated to demonstrate peroxidase activity. Reaction product was present within the endoplasmic reticulum, including the nuclear envelope. Study of several types of control preparations strongly suggest that the antibody-binding material in the endoplasmic reticulum is growth hormone.
AUTOMATIC RECOGNITION OF IRRADIATED CHROMOSOMES
Authors: AGGARWAL RAJ K. , FU K. S.
Published: April 2011

There are essentially three types of aberrations of irradiated chromosomes in a mitotic cell; namely, fragments, rings and dicentrics, which are resulted from breakages and exchanges of chromatids. The occurrence of any of these aberrations is considered to be a very useful indicator of radiation damage at cell level. The large volume of tests required for monitoring health-related radiation damage necessitates an automated system for recognition of these aberrant chromosomes. Such a system, using a cutoff level independent skeletonization approach, has been developed and is discussed in this paper. The test results on 15 abnormal cells are also presented.
NUCLEAR IMAGES OF CELLS IN DIFFERENT FUNCTIONAL STATES
Authors: KIEFER G. , KIEFER R. , MOORE G. W. , SALM R. , SANDRITTER W.
Published: April 2011

To gain further insight into the supramolecular structure and behavior of chromatin, a set of parameters has been developed for the description of cell nuclear images. These images are obtained from Feulgen-stained nuclei in semithin sections. Data are extracted by a scanning microphotometer, printed on paper tape and processed by an electronic computer. The essential principles for the selection of descriptors were that: (a) they should correspond to characteristics that are employed in classical morphology and (b) they should be subject to interpretation in categories of biochemistry or molecular biology insofar as possible. Some of these parameters deal with the size and shape of the nucleus and its deoxyribonucleic acid content or staining behavior. Efforts were made to determine the relative amount of chromatin in the condensed state, because it may be assumed that the degree of condensation correlates with the activity of the chromatin. To demonstrate the power of the method, results from methylthiouracil-activated thyroid nuclei are presented. In stimulated nuclei the diffuse chromatin expands and fills the increasing nuclear volume, while the condensed chromatin retains its degree of condensation, and, probably, does not change in amount.
LIGHT-SCATTERING PATTERNS OF ISOLATED OLIGODENDROGLIA
Authors: MEYER R. A. , HAASE S. F. , PODUSLO S. E. , MCKHANN G. M.
Published: April 2011

A technique for determining the large angle light-scattering patterns of biologic cells has been developed and applied to the measurement of isolated calf oligodendroglia. Preliminary results indicate that light-scattering information can be used to discriminate between oligodendroglia, subcellular debris and red blood cells and also suggest that oligodendroglia can be distinguished from neurons.
AUTOMATED ANALYSIS OF DEOXYRIBONUCLEIC ACID, PROTEIN AND NUCLEAR TO CYTOPLASMIC RELATIONSHIPS IN TUMOR CELLS AND GYNECOLOGIC SPECIMENS
Authors: STEINKAMP J. A. , CRISSMAN H. A.
Published: April 2011

Quantitative two-color fluorescence staining techniques, coupled with flow system multiparameter cell analysis and sorting instrumentation, have been used for rapid, simultaneous determination of deoxyribonucleic acid, protein, nuclear (N) and cytoplasmic (C) diameters and N:C ratios in mammalian tumor cells and human gynecologic specimens. Cells stained in suspension for deoxyribonucleic acid and total protein content, respectively, with propidium iodide (red fluorescence) and fluorescein isothiocyanate (green fluorescence), enter a flow chamber and intersect an argon laser beam which excites cellular fluorescence. Optical sensors measure both red and green fluorescence plus the time duration of each fluorescence signal which is proportional to nuclear and cytoplasmic diameters, respectively. The resulting signals are processed and displayed as frequency distribution histograms using a multichannel pulse height analyzer. Cells are also sorted based on N:C ratios. Illustrative examples of preliminary two-color fluorescence analysis and sorting of mouse squamous tumor cells and human exfoliative vaginal cells are presented.
OPTIMIZATION OF CANCER DETECTION
Authors: MOORE DAN H.
Published: April 2011

A statistical model is developed that describes the population of women who are given a cytologic screening test for cervical cancer. The model is used to determine false positive and false negative rates as a function of (a) the proportion of "positive" cells in women free from cancer and in those with cancer, (b) the number of cells examined and (c) the minimal number of positive cells for a diagnosis of cancer. The model allows estimation of the minimal number of cells that must be examined in order to reduce both the false positive and the false negative rates below some predetermined levels. An expected cost equation is derived which combines the costs of examining each cell with the costs for false positives and false negatives. It is shown how cancer detection can be optimized through the use of this cost equation. The method determines both the maximal permissible cost for examining each cell and the optimal number of cells to examine in order to reduce the over-all expected cost below some predetermined level.
THE APPLICATION OF ARTIFICIAL INTELLIGENCE TECHNIQUES TO BIOLOGIC CELL IDENTIFICATION
Authors: SHAPIRO BRUCE , LEMKIN PETER , LIPKIN LEWIS
Published: April 2011

An interactive system for measuring and classifying cells is discussed. Techniques developed in several artificial intelligence systems are used to provide for a computer-controlled microscope and peripheral hardware a language and control mechanisms to permit the interactive description of cell models. In particular, the system will initially be applied to the analysis of bone marrow smears. An example is given to illustrate the descriptive modeling capability.
THE HISTOCHEMICAL DEMONSTRATION OF O-ACYLATED SIALIC ACID IN GASTROINTESTINAL MUCINS THEIR ASSOCIATION WITH THE POTASSIUM HYDROXIDE-PERIODIC ACID-SCHIFF EFFECT
Authors: CULLING C. F. A. , REID P. E. , CLAY M. G. , DUNN W. L.
Published: April 2011

O-Acylated sialic acids have been demonstrated histochemically in the epithelial mucins of the lower end of the gastrointestinal tract. The potassium hydroxide-periodic acid-Schiff effect has been shown to be due, in most areas, to the presence of these O-acyl esters on the polyhydroxy side chains of sialic acid. The esters have been shown to hinder the removal of the sialic acids by either neuraminidase or acid hydrolysis unless tissues were pretreated with alcoholic potassium hydroxide. It is suggested that substitution at the C4 position is responsible for their neuraminidase resistance.
Determination of the glycogen content in single neutrophil leukocytes using a micromodel of leukocyte glycogen.
Authors: Gahrton G , Olsson I , Dahlqvist A
Published: April 2011

The kinetics of the periodic acid oxidation as part of the periodic acid-Schiff reaction was studied by combined microinterferometry and microspectrophotometry in micromodel systems of liver glycogen and leukocyte glycogen as well as in neutrophil leukocytes. The initial formation of Schiff-positive chromogens was more rapid in neutrophil leukocytes than in liver or leukocyte glycogen. The chromogen formation was, however, practically complete within 60 min in both neutrophil leukocytes and leukocyte glycogen, but this did not appear to be the case in liver glycogen. Differences in the rate of chromogen formation may depend on various factors such as differences in the source and treatment of the glycogen. The complete periodic acid-Schiff reaction appears to be a measure of the glycogen amount in neutrophil leukocytes and the microdroplet system of leukocyte glycogen is considered to be an appropriate model for the estimation of the glycogen amount in single neutrophil leukocytes. A mean value of 13.3 10-12 g glycogen per normal human neutrophil was found.
Bismuth localization within nuclear inclusions by x-ray microanalysis. Effects of accelerating voltage.
Authors: Fowler B A , Goyer R A
Published: April 2011

This report details the localization of bismuth by energy-dispersive x-ray microanalysis within characteristic nuclear inclusion bodies of renal tubular lining cells of rats following excessive exposure to this element. Peak to background ratios and analytical detection sensitivities for bismuth were found to vary for 04, 60 or 80 keV electron accelerating voltages. Optimum peak to background ratios were observed at 40 keV due to lower background generation, while greater detection sensitivities were recorded at 80 keV due to enhanced generation of bismuth characteristic x-rays.
The coating of mouse myocardial cells. A cytochemical electron microscopical study.
Authors: Gros D , Challice C E
Published: April 2011

The coating of mouse myocardial cells has been investigated with a variety of cytochemical methods. The coating of the surface membrane gives a positive reaction with ruthenium red, colloidal thorium, phosphotungstic acid (PTA) at low pH, silver methenamine after periodic oxidation (PA-silver technique) and with silver proteinate after periodic oxidation and thiocarbohydrazide treatment (PA-TCH-silver technique). The coating of the T system gives almost similar results. The nexuses do not react with PTA nor with the PA-silver and PA-TCH-silver techniques, but they are strongly stained with ruthenium red which reveals periodic structures in their gaps. The specificities of the colloidal thorium technique and PAT staining have been tested by chemical treatments (methylation, acetylation, saponification), enzymatic digestions (pronase, trypsin, hyaluronidase, neuraminidase) and carbohydrate extractions (with 0.1 N NaOH and 0.05 M H2SO4). These cytochemical data indicate, considering the specificity of the reactions, that the coating of the membrane surface and the T system contains polyanionic groups. A part of them, at least, would belong to a carbohydrate-containing material (glycoproteins), whereas at the level of nexuses the sugar residues would probably be absent.
N-acetyl-beta-glucosaminidase activity in normal and malignant leukocytes.
Authors: Reed C E , Bennett J M
Published: April 2011

An improved cytochemical method demonstrating N-acetyl-beta-glucosaminidase in peripheral blood and bone marrow leukocytes is described. A significant elevation in enzyme activity in circulating monocytes from patients with solid tumor malignancies was observed. In a large series of cases of acute nonlymphocytic leukemia, elevated levels were found in the vast majority of those leukemias that had a predominant monocytic component identified either morphologically or by standard cytochemical methods. This reaction would appear to be useful as a monocyte marker.
Ultrastructural visualization of virus-induced surface antigens. Comparative studies of three immunohistochemical techniques.
Authors: Morris R E , Fritz R B
Published: April 2011

Three immunohistochemical techniques used for ultrastructural localization of cell surface antigens were compared with respect to ease of reagent preparation and qualitative aspects of cell surface staining. The techniques compared were: (a) ferritin-conjugated antibody, (b) "hybrid" antibody and (c) a modification of the "bridging" technique which employs soluble immune complexes. Used in conjuction with strain MC29 avian oncornavirus-infected chick embryo cells and chicken antiviral immune serum, the results of all three techniques were comparable. In terms of ease of preparation, the "bridging" technique employing soluble complexes was found to be the most satisfactory.
Light microscopic localization of cytochemical reactions in epoxy-embedded material for electron microscopy.
Authors: Russo J , Wells P
Published: April 2011

Tissues from mice were fixed in 1.5% glutaraldehyde, treated for the ultrastructural localization of alkaline phosphatase or Mg++-dependent adenosine triphosphatase, post-fixed in osmium tetroxide, dehydrated and embedded in plastic for electron microscopy. The sites of reaction were visualized in 1-mu plastic sections counterstained with toluidine blue, using a phase contrast microscope. The data show a close correlation between the sites of reaction observed with the phase contrast microscope and the sites studied with the electron microscope. The use of this technique for the study of these phosphatases in normal and pathologic tissues is recommended in order to achieve a high degree of accuracy in selecting a portion of the tissue sample for electron microscopy and to obtain greater resolution in the localization of these enzymes with the light microscope.
Intracellular localization and size of glycogen particles in glycogen synthesized under histochemical conditions.
Authors: Takeuchi T , Sasaki M , Miyayama H , Oyumi M , Miyajima H
Published: April 2011

During the investigation of the histochemical synthesis of glycogen particles from glucose 1-phosphate by the phosphorylase-branching glycosyltransferase system in various tissue cells, it was observed that focal synthesis localized in a certain area of the cytoplasm occurred in some cells. This differed from the usual synthesis in which particles of similar size were synthesized within the cytoplasm. Otherwise, cytoplasmic particles of various size were also synthesized in other cells under the same histochemical condition. The possible significance of the presence of these patterns in glycogen synthesis is discussed.
Abnormal mast cell granules in the beige (Chédiak-Higashi syndrome) mouse.
Authors: Chi E Y , Lagunoff D
Published: April 2011

Mast cells of beige (C57BL/6J) (bg-j/bg-j) mice were examined histochemically and ultrastructurally. Mast cell granules in the beige mice were markedly enlarged and irregular in shape. Granule contents stained uniformly with acidified toluidine blue, but with ruthenium red and Alcian Blue-safranin, two components were evident. The rims of the abnormal granules stained with ruthenium red and with Alcian Blue; the centers of the granules were clear with ruthenium red and stained with safranin. Mast cell granules thus represent another abnormal organelle in the Chédiak-Higashi syndrome.
Adenylate deaminase. A method for its localization in skeletal muscle.
Authors: Nakatsu K
Published: April 2011

A method for light microscopic localization of adenylate deaminase in sections of frozen rat quadriceps muscle is described. The method depends on the hydrolysis of 6-chloropurine ribonucleotide (the 6-chloroanalogue of adenylate) and the trapping of Cl minus by Ag plus. The resulting AgCl precipitate was made visible by exposing the sections to light. After this treatment black deposits about 1 mu in diameter were seen in muscle cells. These observations indicate that adenylate deaminase of rat quadriceps muscle is located at discrete sites within the muscle cells.
A comparative study of horseradish peroxidase conjugates prepared with a one-step and a two-step method.
Authors: Boorsma D M , Kalsbeek G L
Published: April 2011

In this study we compared horseradish peroxidase (HRP)-labeled rabbit antihuman immunoglobulin G (IgG) conjugates, prepared by a one-step and a two-step method. Glutaraldehyde was used as a cross-linking agent. Two methods were used for removing unconjugated HRP: Sephadex G-200 gel chromatography and ammonium sulfate precipitation. The conjugates were characterized immunologically, immunochemically and enzymatically. The immunohistoenzymic properties of the conjugates were tested on unfixed cryostat sections of the skin of patients with chronic discoid lupus erythematosus. The influence of the presence of unconjugated HRP and unconjugated IgG was studied. Optimal results were obtained with conjugates prepared by a two-step method. Removing unconjugated HRPimproved the immunohistoenzymic properties of the conjugates. Conjugated and unconjugated IgG could be separated by Sephadex G-200 gel chromatography.
Comparative ultrahistochemistry of the adepidermal granules of Salmo irideus, Lebistis reticulatus and Hynobiuo tokyoensis. Enzyme digestive experiment for the epoxy-embedded sections;.
Authors: Tachibana T
Published: April 2011

A comparative ultrahistochemical investigation of the adepidermal granules of Salmo irideus, Lebistis reticulatus and Hynobius tokyoensis was carried out using enzyme digestion methods on epoxy-embedded sections. The granules of S. irideus larvae were decomposed by periodic acid, and digested by lipase without periodic acid pretreatmenetection of the granules. The secondary postosmificated granules were digested by lipase as in S; irideus, but complete decomposition by periodic acid was not observed in this experiment; Both periodic acid and lipase changed the shape of the adepidermal granules of H. tokyoensis to suggest partial digestion, but it appeared that these granules show rather stronger resistance to periodic acid and lipase than those of S. irideus. The granules of H. tokyoensis were completely digested when the sections were treated sequentially with phospholipase C, neuraminidase and lipase.
Evaluation of silicon and germanium retention in rat tissues and diatoms during cell and organelle preparation for electron probe microanalysis.
Authors: Mehard C W , Volcani B E
Published: April 2011

Chemical, radiochemical and x-ray microanalysis assays were used to define parameters of silicon (Si) retention during preparation og biologic samples (rat liver, spleen, kidney, lung, diatoms and cell organelles) for x-ray microanalysis, Due to its longer half-life 68-Fe was used in some cases to trace SI. Leaching of Si from cells and organelles by the aqueous preparation media was overcome by use of the freeze-substitution process. Cells were treated with 30% glycerol hypertonic sucrose medium to reduce ice damage. Embedment in Spurr's low viscosity epoxy resin medium caused no apparent Si loss. A semiquantitative evaluation showed 0.5 x 10-8 to 0.3 x 10-17 g detectable Si in isolated rat liver mitochondria in thin sections, which is within the instrument's range of detection. This study indicateds that the presence of Si in the mitochondria is not the rsult of contamination.
Centrifugal cytology. III. The utilization of centrifugal cytology for the preparation of fixed stained dispersions of cells separated by bovine serum albumin bouyant density centrifugation.
Authors: Dunlap L A , Warters R L , Leif R C
Published: April 2011

This paper describes the modification of Centrifugal Cytology for the preparation of permanent, fixed, stained dispensions for both light and scanning electron microscopy of cells which have been isolated on bovine serum albumin (BSA) boyant density gradients. The principal problem with BSA gradient fractions is that the albumin which is present even after dilution is precipitated by the glutaraldehyde fixative. This problem has been solved by the layering of an intermediate D2O solution under the BSA and subsequent removal of the BSA solution and the underlaying with D2O containing glutaraldehyde. A special layering machine facilitates and expedites these operations. This technique has also been applied to BSA-seperated guinea pig and chicken bone marrow cells, as well as Ehrlich ascites tumor cells, hen and human blood cells. The number of celll present in each area of the slide is maintained at a constant value by utlizing a table of dilution factors. This table was generated by a computer program which calculates the concentration of cells present in the rractions and divides it by the number of celll desired.
Variability of the iron, copper and mercury contents of individual red blood cells.
Authors: Colvin J R , Sowden L C , Male R S
Published: April 2011

The relative iron, copper and mercury contents of individual, isolated erythrocytes from eight people were determined by analytical electron microscopy. The variation in iron content between erythrocytes of the same sample is more than six times, for copper content more than tem times and for mercury more than five times. Similar variations were observed for 1-day-old chick defintive erythrocytes and for 4-day-old chick embryo primitive erythrocytes. The range of variation does not depend greatly, it at all, on the age of the erythroyctes or the tissue of origin. There is little or no correlation between the variation of iron contnet and that of copper. The cause of the wide variation of metallic ion contnt among erythroyctes is not yet known.
Inhibition studies of alkaline phosphatase in hard tissue-forming cells.
Authors: Linde A , Magnusson B C
Published: April 2011

The effects of the alkaline phosphatase inhibitors levamisole and R 8231 on p-nitro-phenylphosphatase, inorganic pyrophosphatase and adenosine triphosphatase (ATPase) activities in dentingenically active odontoblasts were studied. The p-nitrophenylphosphatase and inorganic pyrophosphatase activities were inhibited, while 40% of the ATP-splitting enzyme activity remained under the assay condition used. This finding, togeather with earlier studies, indicates that at least two different phosphatase are active at alkaline pH in hard tissue-forming cells; on nonspecific alkaline phosphatase and one specific ATPase. The ATPase activity is uninfluenced by ouabain and ruthenium red and is activated by Ca-2+ ions.
Histochemical fiber typing and staining intensity in cat and rat muscles.
Authors: Schmalbruch H , Kamieniecka Z
Published: April 2011

In the gastrocnemius muscle of cat and rat, staining for oxidative enzymes differentiated three fiber types (A,B,C) and staining for adenosine triphosphate at pH 9.4 differentiated two fiber types (I, II) with a reliability of 90% and 98%, respectively. In cat 96% and in rat 90% of the fibers were typed identically after staining for nicotinamide adenine dinucleotidelinked lactic dehydrogenase (LDH) and succinic dehydrogenase (SDH). When differentiated by staining for LDH, A and B fibers were of type I. IN RAT, 80-90% OF ALL FIBERS WERE OF TYPE 22, COMPPRISING A, B and C fibers. Type I fibers stained for LDH intensely as did C fibers of type II, but stained intermediately for SDH. The degree of staining was measured by photometry. When fibers were stained for LDH, histograms of density showed three peaks corresponding to A, B and C fibers in cat, but only two peaks corresponding to A and C fibers in rat, In cat and rat, the densities of A, B and C fibers belonged to different populations. In soleus muscle of cat and rat stained for LDH, menadione-linked alpha-glycerophosphate dehydrogenase and adenosine triphosphatase at pH 9.4, the degree of staining differed from thatin any type of fiber in gastrocnemius muscle
The efficiency of 14C detection in autoradiographs of semithin plastic sections.
Authors: Housley T L , Fisher D B
Published: April 2011

Sections of uniformly labeled 14C-methylmethacrylate were used to determine the efficiency of 14C detection by Ilford L4 emulsion layers, 0.22 mu in thickness, in light microscope autoradiographs. When efficiency was expressed as a function of radiation dose (i.e., the number of decays/mu2) there was a strong dependency on radiation dose up to about 6 decays/mu2, after which detection efficiency was fairly constant. For most emulsion batches, the detection efficiency was about 5%, but it ranged from 8 to 3%.
Coherent optical processing of cervical cytologic samples.
Authors: Kopp R E , Lisa J , Mendelsohn J , Pernick B , Stone H , Wohlers R
Published: April 2011

Earlier research using a very limited data base gave encouraging results for the automated screening of exfoliated cytologic samples using coherent optical processing techniques to examine individual isolated cells. A more thorough investigation involving a larger data base has confirmed our initial results. This investigation was performed using a specially designed Fourier spectrum analyzer and a solid state optical detector array. An analysis was made to determine the performance of a screening system using such a cell-by-cell discriminating device. This analysis indicated that less than 20,000 cells would have to be examined to obtain a system performance level of 1% false negative and 10% false positive error rates with a 1% probability of occurrence of malignant cells in a malignant sample. This performance figure was inferred from measured statistical performance characteristics of a laboratory cell-by-cell screening device using optically generated Fourier transfrom techniques for cell discrimination. The performance of the system was shown to be much more sensitive to cell-by-cell false error rates than false negative error rates. It was also found that the majority of false positive errors were due to misclassifying parabasal cells as malignant. By eliminating parabasal cells, which comprised more than 25% of our normal cell data base, the number of cells needed to be screened dropped by an order of magnitude. It was also shown that there is an inverse quadratic relationship between the percentage of malignant cells in a malignant sample and the number of cells that must be screened to achieve any desired system performance.
Automatic detection and localization of sister chromatid exchanges.
Authors: Zack G W , Spriet J A , Latt S A , Granlund G H , Young I T
Published: April 2011

Sister chromatids of human metaphase chromsomes from cells which have replicated twice in medium containing 5-bromodeoxyuridine exhibit unequal fluorescence when stained with the dye 33258 Hoechst. Sister chromatid exchanges occurring in these chromosomes are apparent as interchanges of brightly and dully fluorescing chromatids. A technique for detecting such exchanges by computer analysis of chromsome images has been developed and found to campare favorably with manual methods. The exchanges have been localized in the context of quinacrine banding patterns.