bluuebunny/crossref_metadata_2025_split huggingface dataset:
Signal Amplification in the Detection of Single-copy DNA and RNA by Enzyme-catalyzed Deposition (CARD) of the Novel Fluorescent Reporter Substrate Cy3.29-Tyramide
Authors: Schmidt Brigitte F. , Chao Jean , Zhu Zhengrong , DeBiasio Robin L. , Fisher Gregory 
Published: April 2011

We demonstrate that the CAtalyzed Reporter Deposition method (CARD), utilizing the novel fluorescent reporter Cy3.29-tyramide, is successful in the Fluorescent In Situ Hybridization (FISH) detection of RNA and single-copy DNA. Histone 4 expression is detected in RNA extracts of S-phase, synchronized HeLa cells by dot-blot analysis. Gene expression of histone 4 in HeLa cells is demonstrated by FISH via CARD, utilizing oligonucleotide probes. Fluorescence intensity measurements on CARD-amplified histone 4 RNA detection showed (a) a 25-fold amplification of the signal brightness by biotinylated oligonucleotide probes and (b) a sixfold amplification of the signal brightness by horseradish peroxidase (HRP)-labeled histone 4 probes vs the directly stained control. The sensitivity of the CARD method is demonstrated by the FISH detection of single-copy DNA on human corneal fibroblast and HeLa S3 interphase nuclei. Chromosomal localization of the single copy DNA is demonstrated on HeLa S3 metaphase chromosome spreads. (J Histochem Cytochem 45:365–373, 1997)
Loss of Heterozygosity of Tumor Suppressor Genes (p16, Rb, E‐cadherin, p53) in Hypopharynx Squamous Cell Carcinoma
Authors: Lee Sang‐Hyuk , Lee Nam‐Hoon , Jin Sung‐Min , Rho Young‐Soo , Jo Sung‐Jin 
Published: April 2011

ObjectiveMicrosatellite alterations, especially those that cause loss of heterozygosity (LOH), have recently been postulated as a novel mechanism of carcinogenesis and a useful prognostic factor in many kinds of malignant tumors. However, few studies have focused on a specific site, hypopharynx. The aim of this study was to evaluate the relationship between LOH and hypopharyngeal squamous cell carcinoma (HPSCC).Study DesignLaboratory‐based study.SettingIntegrated health care system.Subjects and MethodsMatched normal and cancerous tissues from 30 patients with HPSCC were examined for LOH in 4 tumor suppressor genes (TSGs) (p16, Rb, E‐cadherin, and p53) at loci 9p21, 13q21, 6q22, and 17p13, respectively, using microsatellite markers amplified by polymerase chain reaction. The results for each loci were compared with clinicopathological features.ResultsAmong the 30 cases, 26 (86.7%) exhibited LOH, with the most common alteration being LOH at p53 (52.6%). Significantly higher rates of LOH detection were seen in Rb, p53, and the LOH‐high group (cases where 2 or more loci with LOH were found) in cases of lymph node metastasis. Compared with stage I and II carcinoma, tumors of stages III and IV had significantly higher frequencies of LOH in Rb, p53, and the LOH‐high group. However, the presence of LOH was not significantly correlated with survival.ConclusionThese results suggest that LOH in TSGs such as Rb and p53 may contribute to the development and progression of HPSCC. The presence of LOH in the primary tumor may also be predictive of lymph node metastasis.
Third Windows as a Cause of Failure in Hearing Gain after Exploratory Tympanotomy
Authors: Kim Sang Cheol , Lee Won‐Sang , Kim Minbum , Jeon Ju Hyun , Kim Jinna , Choi Jae Young 
Published: April 2011

ObjectiveExploratory tympanotomy (ET) can be performed selectively on patients who have air‐bone gaps (ABGs) without any apparent external or middle ear abnormalities for diagnosis and treatment of their conductive hearing loss. However, surgeons cannot always find middle ear problems. It has recently been reported that pathologic third windows in inner ears can cause ABGs. Therefore, the authors conducted this study to evaluate the inner ear anatomical problems of patients who had undergone ET.Study DesignCase series with chart review.SettingTertiary center.Subjects and MethodsThe authors included 51 consecutive patients with ABGs ≥20 dB who had undergone ET at Yonsei University Severance Hospital between 2005 and 2009. In the preoperative evaluations, all 51 patients showed normal drums on otoscopy, and no abnormal middle ear problems that could cause ABGs were detected on temporal bone computed tomography (TBCT). The authors retrospectively reevaluated the hearing results and the causes of their conductive hearing loss, including the inner ear problems, through a review of their TBCT.ResultsSix of the 51 patients were found to have inner ear abnormalities with possible pathologic third windows. Four had enlarged vestibular aqueducts (EVAs), and 2 had superior semicircular canal dehiscence (SSCD). All patients with pathologic third windows failed to improve their hearing.ConclusionAbout 12% of the patients had inner ear anomalies, which could possibly cause ABGs. Inner ear conductive components might lead to frustrating results. Therefore, surgeons must take inner ear conductive components into account before attempting ET in patients with inner ear anomalies.
THE ACTION OF FORMALDEHYDE ON RAT BRAIN LIPIDS
Authors: DEIERKAUF F. A. , HESLINGA F. J. M. 
Published: April 2011

The influence of formaldehyde on rat brain lipid was investigated by comparing the analysis of lipids from fresh rat brain with the analysis from rat brains fixed for 110 days in a formaldehyde solution. A quantitative analysis of the lipids using column and paper chromatography is described. Sphingomyclin, cerebrosides and sulphatides were not attacked. Only a slight diminution of phosphatidylserine was indicated. Phosphoinositides remain fairly constant. Of lecithin about 25% was converted to lyso-lecithin. The phosphatidylethanolamine and lyso-phosphatidylethanolamine fractions suffered a marked diminution and the reaction with formaldehyde was noticeable in a change of chromatographic behaviour. The fraction containing cholesterol, cholesterol esters, glycerides, and fatty acids increased, the increase being correlated with the decrease of other fractions by the formation of lyso compounds. After storing tissues for months in formaldehyde solutions a reliable analysis of the lipid content is still possible.
THE REACTION OF OSMIUM TETROXIDE WITH LECITHIN: APPLICATION OF STAINING PROCEDURES
Authors: RIEMERSMA JELLE C. , BOOIJ HEIN L. 
Published: April 2011

To obtain additional insight into the chemical reactions occuring furing fixation of tissue lipids by osmium tetroxide, the reaction between osmium tetroxide and lecithin has been studied. Procedures have been developed whereby quantitative information may be obtained about the nature of this reaction. One series of experiments was designed to arrive at the stoichiometry of the reaction and its time-course. To this end air-dried lecithin spots on cover glasses or on filter paper were exposed to oxmium tetroxide vapor. The weight increase of the lecithin spots was determined, and related to time of exposure to osmium tetroxide and to the quantity of lecithin present. Under the conditions chosen the binding of osmium tetroxide rapidly reaches a limiting value; thus the reaction appears to proceed to a definite end-point. The number of double bonds in the lecithin molecule, as determined by gas chromatography, corresponds with the number of moles of osmium tetroxide bound per mole of lecithin. This supplements earlier spectrophotometric findings, and confirms part of the reaction scheme proposed by Criegee for the interaction between osmium tetroxide and unsaturated compounds generally. The chemical properties of lecithin, and of other polar lipids, are determined not only by the presence of unsaturated linkages but also by the presence of an ampho-ionic group. Earlier research has demonstrated that this group is responsible for the staining properties of lecithin with anionic dyes, for instance, Brilliant Scarlet 3 R (Ponceau 4R or 4RS, Colour Index No. 16255). In this paper evidence is presented to show that the stainability of lecithin spots by Brilliant Searlet 3 R, under specified conditions, decreases markedly with increasing time of exposure to osmium tetroxide. It appears likely that as a result of the reaction with osmium tetroxide the lecithin polar group has changed properties. After studying the staining properties of lecithin towards an anionic dye, in relation to time of exposure to osmium tetroxide, its staining capacity for a representative cationic dye was similarly studied. Evidence is presented demonstrating an increased binding capacity of exposed lecithin spots to Brilliant Green (Colour Index no 42040). After considering a few possible interpretations we suggest that the simplest hypothesis to account for this phenomenon is a partial removal of the positive charge of lecithin, whereby the negatively charged groups are freed for interaction with positive dye molecules.
OXIDATIVE AND HYDROLYTIC ENZYMES OF HOMOZYGOUS DYSTROPHIC AND HETEROZYGOUS MUSCLE OF THE HOUSE MOUSE
Authors: FENNELL RICHARD A. , WEST WILLIAM T. 
Published: April 2011

Histochemical procedures were used for the identification of oxidative and hydrolytic enzymes of homozygous dystrophic (genotype, dydy) and heterozygous (genotype, Dydy) muscles of the house mouse. In adult heterozygous muscle and muscle of a Swiss albino mouse there was usually a reciprocal relationship between phosphorylase and lactate dehydrogenase, i.e., weak lactate dehydrogenase reactions in the majority of fibers of the vastus lateralis and a stronger phosphorylase reaction. Weak to strong lactate dehydrogenase reactions were identified in fibers of the rectus femoris. In heterozygous muscle of young mice 20 days of age, a reciprocal relationship between lactate dehydrogenase and phosphorylase was not observed. Reciprocity between phosphorylase and lactate dehydrogenase was not always evident in fibers of homozygous muscles, e.g., groups of homozygous fibers frequently exhibited both strong phosphorylase and lactate dehydrogenase reactions whereas other fibers reacted weakly for phosphorylase or not at all and strongly for lactate dehydrogenase. Small fibers reacted either weakly or strongly for dehydrogenases. One of the striking features which homozygous fibers usually showed was uniformly strong dehydrogenase reactions (e.g. lactate, glutamate, succinate) and diphosphopyridine nucleotide linked diaphorase reactions. Strong glucose-6-phosphate dehydrogenase reactions were frequently identified in homozygous dystrophic fibers whereas weak reactions or no reactions at all were seems in heterozygous fibers. Atrophic fibers of homozygous dystrophic muscles exhibited intense indoxyl acetate esterase and acid phosphatase reactions. Weak adenosine triphosphatase reactions were seen in fibers of both heterozygous and homozygous dystrophic muscle, and strong reactions in connective tissues and blood vessels. Interfibrillar connective tissue of homozygous muscle was abundant and exhibited intense acid phosphatase reactions, whereas connective tissues of heterozygous muscle were less concentrated and exhibited weak acid phosphatase reactions.
THE ROLE OF IRON IN THE MECHANISM OF EXPERIMENTAL CALCIFICATION
Authors: GABBIANI G. , TUCHWEBER B. 
Published: April 2011

Calcification of different organs in the rat, induced by challenge with various iron compounds after sensitization with dihydrotachysterol, was studied histochemically. Iron is demonstrable in the target organs before calcification but apparently, in the described experimental conditions, it can attract calcium only when it is extracellular. The possible relationship between ferrification and calcium deposition is briefly discussed.
ELECTRON MICROSCOPIC CYTOCHEMISTRY OF MELANOSOMES AND MITOCHONDRIA
Authors: MISHIMA YUTAKA 
Published: April 2011

The electron histochemical properties characteristic of melanosomes as distinguished from mitochondria are presented. Our modified ammoniated silver nitrate reaction for electron microscopy reveals an essential chemical property of melanosomes which is also possessed by melanin granules, but is not possessed by any mitochondria. This property is the reduction of ammoniated silver nitrate to the metallic state. Furthermore, melanosomes, and not mitochondria, are the only organelles which acquire the appearance of typically mature melanin granules after the electron microscopic dopa reaction.
A METHOD FOR THE CYTOCHEMICAL DEMONSTRATION OF SODIUM-ACTIVATED ADENOSINE TRIPHOSPHATASE
Authors: MCCLURKIN lOLA T. 
Published: April 2011

(1) In the histochemical method for adenosine triphosphatase, when magnesium and ATP are in equivalent concentrations, and sodium is present in the medium, staining occurs in the tissues. When sodium is omitted from this medium some structures—previously stained in the presence of sodium—fail to stain. (2) Ouabain inhibits staining when magnesium and ATP are equivalent and sodium is present in the medium. (3) The effects of calcium, potassium and sodium on staining with the proposed medium parallel the effects of these ions on activity of sodium-activated ATPase in vitro. (4) When magnesium ions and ATP are not equivalent in the substrate, sodium and ouabain fail to affect staining. (5) Certain tissue structures—such as the axonic nerve ending—which have been shown to contain sodium-activated ATPase in vitro, stain in the presence of sodium when magnesium and ATP are equivalent, but fail to stain when sodium is omitted from this medium.
INDOPHENOL OXIDASE AND PEROXIDASE ACTIVITIES IN MAMMALIAN ERYTHROCYTES
Authors: SIEGEL ISRAEL 
Published: April 2011

Cytochrome oxidase and peroxidase activities in mammalian red cells are suggested by G- and M-Nadi reactions in rabbit, mouse, and human erythrocytes. Peroxidase activity was present in both the hemoglobin and the nonhemoglobin constituents of the red cells. Removal of oxygen stopped the G-Nadi reaction but not the M-Nadi reaction. Potassium cyanide and sodium azide at a concentration of 10–3-10–4 M partially inhibited G- and M-Nadi reactions. Microscopic examination of reacting cells revealed accumulation of reaction products at the cell periphery. This was not evident when the cells were incubated in a suspension of indophenol blue. Mammalian erythrocytes reduced the tetrazolium salt nitro-BT. Reduction was augmented by removal of oxygen and by cytochrome oxidase inhibitors. Accumulation of formazan at the surface of reacting cells was evident upon microscopic examination. The Nadi reactions were similar whether Ehrlich ascites tumor cells or stroma of hemolyzed erythrocytes were used. Reactions of stroma or whole erythrocytes with nitro-BT were essentially similar. Fixation or heating slowed or abolished all the reactions described.
Medical and pharmacological direct costs of a 9-week smoking cessation programme
Authors: Wasserfallen Jean-Blaise , Digon Patricia , Cornuz Jacques 
Published: March 2011

Background: Medical and pharmacological direct costs of cigarette smoking cessation programmes are not covered by health insurance in several countries despite documented cost-effectiveness. Design: prospective cost identification study of a 9-week programme in Switzerland. Methods: A total of 481 smokers were followed-up for 9 weeks. Socio-demographic characteristics, number of outpatient visits, dosage and frequency of use of nicotine replacement therapy (NRT) as well as date of relapse were prospectively collected. Individual cost of care until relapse or programme end as well as cost per week of follow-up were computed. Comparisons were carried out between the groups with or without relapse at the end of the programme. Results: Of the 209 men and 272 women included, 347 patients (72%) finished the programme. Among them, 240 patients (70%) succeeded in quitting and 107 patients (30%) relapsed. As compared with the group relapsing by the end of the programme, the group succeeding in quitting was more often living in a couple (68% vs. 55%, p = 0.029). Their mean weekly costs of visits were higher (CHF 81.2 ± 6.1 vs. 78.4 ± 7.6, p = 0.001), while their mean weekly costs for NRT were similar (CHF 24.2 ± 12.6 vs. 25.4 ± 15.9, p = 0.711). Mean total costs per week were similar (CHF 105.4 ± 15.4 vs. 103.8 ± 19.4, p = 0.252). More intensive NRT at week 4 increased the probability not to relapse at the end of the programme. Conclusions: Over 9 weeks, medical and pharmacological costs of stopping smoking are low. Good medical and social support as well as adequate NRT seem to play a role in successful quitting.
Gender differences in the extent and timing of the reflected arterial waveform in the presence of coronary artery disease
Authors: Crilly Michael A , Williams David J , Clark Hazel J , Hillis Graham S , Lee Amanda J 
Published: April 2011

Background: The non-invasive assessment of arterial dysfunction may improve cardiovascular (CV) risk assessment. We studied the relationship of the reflected wave transit time (RWT) and augmentation index (AIX), both derived from pulse wave analysis (PWA), in the presence/absence of coronary artery disease (CAD), and compared PWA with the ankle-brachial index (ABI). Methods: A trained research nurse measured PWA (radial applanation tonometry, SphygmoCor device) and ABI (hand-held Doppler) in a consecutive series of fasted patients resting supine prior to elective coronary angiography. Measurements were undertaken blind to other clinical information. Mean differences in RWT, AIX, and ABI in the presence of CAD were adjusted for age, height, mean BP, fasting cholesterol, ever smoked, and treated hypertension using multiple linear regression. Results: We recruited 125 patients (49 women) with a mean age of 65 years, total cholesterol 4.4 mmol/l, BP 136/78, current smokers 22%, and previous myocardial infraction 30%. A statistically significant interaction between sex and CAD was present for both RWT ( p = 0.003) and AIX ( p = 0.03). No interaction was demonstrated for ABI ( p = 0.21). Mean differences for men and women in the presence/absence of CAD were: RWT −10.1 vs. +5.2 milliseconds; AIX +1.2 vs. −5.4; ABI −0.02 vs. −0.10. Male and female area under receiver operating characteristic curves for CAD detection differed for RWT (0.33 vs. 0.67) and AIX (0.62 vs. 0.36), but were similar for ABI (0.40 vs. 0.34). Conclusion: The timing and extent of arterial wave reflections in the presence of CAD may differ in men and women.
COMPARATIVE HISTOCHEMICAL AND QUANTITATIVE STUDY OF ADRENAL AND KIDNEY TISSUE BY TETRAZOLIUM TECHNIQUE
Authors: CASCARANO J. , ZWEIFACH B. W. 
Published: April 2011

A study was made of the influence of experimental variables, including electrolyte balance, pH, tonicity, length of incubation and dimensions of tissue slice, on the in vitro reduction of tetrazolium. The most discriminating approach included examination of both the histochemical and quantitative expressions of the deposition of formazan. Some of the limitations of neotetrazolium (NT) and 2,3,5 triphenyl tetrazolium chloride (TTC) are discussed, NT being less suitable for quantitative estimates of reduction values.
IDENTIFICATION OF CYTOPLASMIC BASOPHILIA (RIBONUCLEIC ACID) BY FLUORESCENCE MICROSCOPY
Authors: VON BERTALANFFY LUDWIG , BICKIS IVAR 
Published: April 2011

1. The fluorochrome, acridin orange (AO), allows identification of basophilic cytoplasmic inclusions in the supravital state. 2. The fluorescence picture of hepatic cells after treatment with AO shows the hyaloplasm faint gray-green; flake-like or granular structures in the cytoplasm, bright red; the nuclei, with chromatin structures, green; the central part of the nucleoli red, their periphery green. 3. The red-fluorescent cytoplasmic inclusions correspond to those observed with the toluidine-blue technique, and shown, by the RNase test, to consist mainly of RNA. The same is demonstrated by the changes of AO fluorescence after application of RNase to fresh or fixed tissues. 4. The fluorescence picture given by other cell elements and tissues in the rat liver is described. 5. Changes in the AO fluorescence of liver tissue with varying pH, dye concentration, staining time, action of salts, prolonged U.V. illumination, fixation, as well as the fluorescence of some biologically important substances (RNA, DNA, heparin, albumin, glycogen), treated with AO, are indicated. 6. The advantages and limitations of the method, and the theories of AO staining are discussed. 7. As compared to conventional histochemical methods, the applicability to living or supravital tissues of the fluorochrome technique decreases errors due to fixation, dehydration, embedding and related procedures, and the low concentrations applied in fluorochrome staining further safeguard against histochemical artifacts.
STUDIES IN HISTOCHEMISTRY: XLIII. THE DESIGN AND USE OF IMPROVED APPARATUS FOR THE PREPARATION AND FREEZING-DRYING OF FRESH-FROZEN SECTIONS OF TISSUE
Authors: GRUNBAUM BENJAMIN W. , GEARY JOHN R. , GLICK DAVID 
Published: April 2011

The design and use of three pieces of apparatus which facilitate and speed sampling of fresh frozen tissue, sectioning, and subsequent handling of sections for freezing-drying or other procedures were described. These are: A tissue borer that produces cylinders of tissue of an accurately determined diameter with minimum distortion, and provides a mounting support for microtoming that permits even thin samples to be sectioned without the contamination of a supporting medium. An alignment device that is used to set the borer with its tissue in the most suitable position in the head of the microtome. A section holder for freeze-drying and handling cut sections conveniently without damage to them. Tests have shown that the use of this equipment mechanizes the procedure to the point where subjective factors are largely reduced and accurate sectioning with little variability can be achieved with relative ease.
A COMPARISON BETWEEN THE HISTOCHEMICAL DEMONSTRATION OF NON-SPECIFIC ESTERASE ACTIVITY BY 5-BROMOINDOXYL ACETATE, α-NAPHTHYL ACETATE AND NAPHTHOL AS ACETATE
Authors: PEARSON BJARNE , DEFENDI VITTORIO 
Published: April 2011

5-Bromoindoxyl acetate was used as a substrate for the histochemical demonstration of esterases. The pH optimum for this salt as used in this study was approximately 5. This differed from the optimum when α-naphthyl and naphthol AS acetates were used, which showed an optimum as determined by color reaction of between 7.3 and 8.4. This latter may be an artifact due to non-enzymatic hydrolysis. Naphthol binding may also vitiate the results. If these hydrolytic artifacts could be eliminated, the true optimum may be of a lower order. No hydrolysis or diffusion artifacts occur in any pH range with the 5-bromoindoxyl acetate. This may occur with the unsubstituted indoxyl acetate. The esterase reactions in various tissues following the use of 5-bromoindoxyl acetate was similar to the α-naphthyl and naphthol AS acetates with the exception of the large intestine which was negative with the 5-bromoindoxyl acetate. The reaction with 5-bromoindoxyl acetate was enhanced by taurocholate in the pancreas and inhibited in the kidney, liver and intestine. This seems to indicate that 5-bromoindoxyl acetate is hydrolyzed by both lipases and esterases from the data of this study. Because of the fine and uniform granularity of the resulting 5,5' dibromoindigo crystals, this substrate should be valuable in the study of esterases.