Search

Purpose: Ghrelin is an orexigenic peptide hormone secreted in times of stress and hunger. It is deeply involved in the regulation of metabolism and energy homeostasis, promoting energy intake and inhibiting energy expenditure on a metabolic level. In this regard, it has in many ways antagonistic effect on the thyroid hormones, which increase metabolism and thus energy expenditure. While there is reasonable evidence of a negative association between ghrelin and hormones of the hypothalamic-pituitary-thyroid (HPT-) axis from studies in patients with thyroid dysfunction and small intervention studies, large-scale studies in healthy subjects are lacking. Therefore, we studied the relationship between total ghrelin serum levels and serum levels of the thyroid hormones in a large sample of euthyroid subjects. Methods: Total ghrelin, thyroid-stimulating hormone (TSH), free thyroxine (fT4) and free triiodothyronine (fT3) were determined after an overnight fast in 1666 subjects participating in a population-based cross-sectional study (‘LIFE’) including 10,000 adults. 1012 subjects were included in this analysis. Multiple linear regression analyses were performed. Results: FT3 was negatively associated with serum ghrelin; total sample: β= − 0.0001, p< 0.001; men: β= − 0.0002, p= 0.013; women: β= − 0.0001, p= 0.010, adjusted for age, BMI, alcohol consumption, serum levels of TSH and fT4 and smoking status. No associations were found between ghrelin serum levels and serum levels of fT4 or TSH. Conclusion: This is to date the largest study investigating the relationship between total serum ghrelin and thyroid hormones. The results point to a complex interaction and should initiate further research.

Four fuchsin analogues (Pararosaniline, Rosaniline, Magenta II and New Fuchsin) usually found in Basic Fuchsin have been applied as chemically pure dyes to the Feulgen-technique. Total nuclear absorption and wavelength of the absorption maximum were measured by microspectrophotometry in Feulgen stained cytological and plastic embedded histological liver samples, and in lymphocyte nuclei in human peripheral blood smears; absorption spectra of Feulgen stained DNA-polyacrylamide films were determined by spectrophotometry. The grey value distribution of tetraploid liver cell nuclei was calculated with an image analyzer. The staining characteristics of the pure dyes were compared to commercial fuchsin samples from various suppliers. Reverse phase thin layer chromatography was used for characterization and qualitative separation of commercial batches.

The aim of this study was to investigate the staining characteristics of Victoria Blue B in alcohol solutions. Cytological specimens (liver and spleen tissue imprints, blood smears) were stained with methanol solutions of commercially available Victoria Blue B-Cl and with pure Victoria Blue B-BF4. The dye concentration, staining time, and protone concentration of the dye solution were varied. The dye solutions were characterized using spectrophotometry and thin-layer chromatography. Cytophotometry and image analysis were used to quantitate the staining pattern of cell nuclei. Feulgen-stained slides were used as controls. Victoria Blue B-BF4 gave excellent nuclear staining exhibiting a quantitative dye-substrate relationship, whereas commercial dyes resulted in lower staining intensity and less distinct nuclear texture. Dye concentration and staining time were, over wide ranges, not of critical importance for the quality of the staining. Under certain staining conditions, only cell nuclei were stained, with the background remaining completely unstained. We presume that, in alcohol solutions, Victoria Blue dye binds as a neutral dye molecule in conjunction with its anion. Victoria Blue B-BF4 staining provides a simple and reproducible staining technique for cytology which is suitable for use in automated cell-pattern recognition.

A method is described for the purification of the dye azure B in quantities sufficient for biological staining experiments on a larger scale. The method is based on the use of column chromatography. Two columns are employed. In column A with silica gel as adsorbent the azure B fraction is isolated from a suitable substrate ('technical' azure B gained by a modification of Bernthsen's synthesis of methylene blue, or plychrome methylene blue) using an acetate-formate mixture as eluent. In column B, on an Amberlite polyineric adsorbent (XAD-2) the acetate-formate anions are exchanged for chloride. Regeneration of both columns is possible: KMnO4, Na2S2O4 and water are run through column A, 5% NaOH, methanol and water through column B. Purification of azure B on economic terms is thus attained. The opinion is expressed that this method is also applicable to the purification of other cationic dyes.

A method is described for the separation of azure A from commezcial samples of polychrome methylene blue. Up to 300 mg of the pure dye may be isolated in this way. The method is based on chromatography using columns 90 cm high, 7 cm in diameter, loaded with 3 g of polychrome methylene blue. The absorbent is silica gel, the eluent a mixture of acetic and formic acid.Poor solubility of the dye acetate in water necessitates dissociation of the acetate by alkalinization and subsequent conversion of the dye to the chloride with diluted Ha. Demethylation that occasionally occurs during this procedure is negligible. Pure azure A does not spontaneously demethylate under ordinary conditions.

Mouse fibroblasts were grown in a ‘deficient’ medium consisting solely of Hank's salt solution. Treatment of cells after 48 hours in deficient medium with the cationic dye, mepacrine, resulted in an altered formation of lysosomes. Cells grown in full medium form dense, multi-vesiculated lysosomes in response to mepacrine. Cells grown in deficient medium form large electron lucent vacuoles containing peripherally located dense globules. The significance of the observations in relation to the deficiency of the medium is discussed.

Summary The Romanowsky-Giemsa staining (RG staining) has been studied by means of microspectrophotometry using various staining conditions. As cell material we employed in our model experiments mouse fibroblasts, LM cells. They show a distinct Romanowsky-Giemsa staining pattern. The RG staining was performed with the chemical pure dye stuffs azure B and eosin Y. In addition we stained the cells separately with azure B or eosin Y. Staining parameters were pH value, dye concentration, staining time etc. Besides normal LM cells we also studied cells after RNA or DNA digestion. The spectra of the various cell species were measured with a self constructed microspectrophotometer by photon counting technique. The optical ray pass and the diagramm of electronics are briefly discussed.

The need for the standardization of reagents and methods used in the histology laboratory is demonstrated. After definitions of dyes, stains, and chromogenic reagents, existing standards and standards organizations are discussed. This is followed by practical instructions on how to standardize dyes and stains through the preparation of reference materials and the development of chromatographic methods. An overview is presented of the problems concerned with standardization of the Romanowsky-Giemsa stain for cytological and histological application. Finally, the problem of how to convince routine dye and stain users of the need for standardization in their histology laboratories is discussed.

The stability of Azure B and Eosin Y in stock solutions of the individual compounds as well as in mixtures of the two dyes was studied. The purpose of the study of these two essential constituents of the Romanowsky Giemsa stain, commonly used in cytology and histology, was to select a stable mixture as a definitive stock solution. Two specific high performance liquid chromatographic methods were used to monitor qualitative and quantitative changes in solutions. Several parameters influencing the stability of Azure B were examined e.g. the type of counter ion the presence of Eosin Y and the type of solvent used. The second part focused on the stability of Eosin Y in mixtures with different cationic dyes submitted to high temperatures. In conclusion, an Azure B SCN-Eosin Y acid mixture in dimethylsulphoxide (concentrations 0.75% and 0.12%, respectively) was selected as being the most appropriate composition of a stock solution for the Romanowsky Giemsa stain.

The chances of Romanowsky-Giemsa (RG) staining becorning a reliable and useful histochemical procedure are reviewed, based on the now proven fact that RG staining requires two dyes only, namely, cationic Azure B and anionic Eosin Y. These two dyes differe from otherwise similar dye combinations in that they give, on distinct biological substrates, one additional colour, purple, which cannot be obtained by the use of either dye alone. The purple colour characterizes the Romanowsky-Giemsa effect (RGE), which is the essential feature of RG staining. Consideration is given to the physico-chemical and morphological implications of RGE. Of primary importance is the nature of the biological substrates where RGE occurs, and also of those where it has never been observed. The way substrates react to RG stains largely depends on the kind of pretreatment they have received; for instance, alcoholic fixation preserves RGE but formaldehyde may inhibit it. Physico-chemical factors are considered which, by altering either the biological substrates or the composition of the staining solutions, may modify the RG staining pattern. This review also serves as an introduction for a series of experimental papers that will follow and which are intended to consolidate the basis of RG staining, a method which holds much promise as a useful histochemical tool.

After incorporation into a polyacrylamide matrix, the biopolymers DNA, RNA, heparin, hyaluronic acid, collagen and the synthetic polymers poly(U) and poly(A, U) were stained with the pure thiazine dyes, Methylene Blue, the Azures and Thionin alone and combined with Eosin Y. Satisfactory spectrophotometric agreement was obtained between the staining reactions of the biopolymers in the artificial matrix and those in their natural surroundings. This was especially true with respect to the specificity of the Azure B-Eosin Y dye-pair, which is based on the generation, on suitable substrates, of a purple colour, the Romanowsky-Giemsa effect (RGE), with an absorbance maximum near 550 nm. In the model experiments, DNA, heparin, hyaluronic acid and collagen were found to be RGE-positive and poly(U), poly(A, U) and RNA RGE-negative. A theory of RGE is proposed which complies with the new and earlier observations: after saturation of available anionic binding sites and aggregate formation by Azure B, electron donor acceptor complexes are formed between Eosin Y and Azure B via hydrogen-bridge formation of the aminosubstituent proton of Azure B and between Eosin Y and the biopolymer surface. Charge-transfer complex formation may also account for the qualitative identity of Azure B-Eosin Y and Azure A-Eosin Y spectra of substrates, which are coloured purple. Quantitatively, Azure A-Eosin Y is less efficient in giving RGE. The generation of RGE is time-dependent. Equilibrium staining is attained after about 120 h. The implications of the results for the biological application of Romanowsky-Giemsa staining are discussed briefly.

With respect to their ability to produce Heinz bodies (HB) in vitro and in vivo in human and mice erythrocytes, a selection of well known cationic vital stains was examined. The stains were compared with phenylhydrazine. After oxazin and phenothiazine dyes, HB developed later than after phenylhydrazine, were detectable for longer and showed a different structure.

Es wurde die Sekundärfluorescenz der Erythrocyten nach AO-Fluorochromierung und zugleich im Phasenkontrast die Wirkung dieser Acridinorange-Fluorochromierung auf die Erythrocyten untersucht.

Es wurde über die Acridinorange-Vitalfluorochromierung des Mäuseasciteskarzinoms unter besonderer Berücksichtigung der intraplasmatischen Speicherung des Farbstoffs in granulärer Form berichtet.

Ausbreitung auf Oberflächen (Etalement), amöboide Bewegungen und Pinocytose als weitgehend spezifische Leistungen der Phagocyten werden untersucht, wobei gemeinsame Voraussetzungen, Dynamik, Endstadien, Verschiedenheiten und gegenseitige Beziehungen dieser differenzierten Zelleistungen besonders berücksichtigt werden.

The fine structure of the substantia granulo-filamentosa of reticulocytes was studied after the use of various dyes. The cytoplasm of unstained reticulocytes contains mitochondria, vacuoles with varying contents, ferritin particles, and numerous polyribosomes. The effects of certain dyes on the nature, distribution, and ultrastructure of the substantia granulofilamentosa are characteristic, and within limits specific for the compound used. Neutral red, 9-aminoacridine, and toluylen blue cause the coarsest precipitates accompanied by a depletion of polyribosomes. The precipitates following the use of brilliant cresyl blue, acridine orange, toluidine blue, and crystal violet are more delicate. Pseudoisocyanin has only slight effects on cytoplasmic detail. Without doubt the essential elements forming the substantia granulofilamentosa are polyribosomes. The stimuli causing their aggregation are unknown; certain theoretical aspects of the problem are discussed.

Zusammenfassung Durch Applikation von 3,4-Benzpyren auf die Mäusehaut wurden in bekannter Weise Papillome und schließlich Carcinome erzeugt. Während die Plattenepithelzellen der Haut nach Vitalfärbung mit Acridinorange das Phänomen der granulären Farbstoffspeicherung nicht erkennen ließen, war dies in Zellen der Fall, die aus Benzpyren behandelten Hautbereichen stammten, welche makroskopisch und histologisch als Carcinome zu bezeichnen waren. Auch morphologisch imponierten diese Zellen als Tumorzellen. Der Befund wird kurz diskutiert.

Zusammenfassung Nach Exposition von Retikulozyten gegenüber kationischen Farbstoffen verschiedener chemischer Struktur werden morphologische Unterschiede in der Art der Präzipitation der Sustantia granulofilamentosa nachweisbar. So sind nach Einwirkung von bestimment Acridinderivaten zahlreiche schollig-kompakte, nach Anwendung von Thiazinverbindungen eher granuläre Niederschläge nachzuweisen. Triphenylmethanfarbstoffe sind zur lichtmikroskopischen Darstellung der Substantia granulofilamentosa in Froscherythrozyten nur bedingt geeignet. — Auch Hinweise auf quantitative Wirksamkeitsdifferenzen liegen vor. — Die Ausfällung der Substantia granulofilamentosa gelang mit einzelnen kationischen Substanzen ohne Farbstoffcharater (Alkaloide).

Zusammenfassung Unter Anwendung verschiedener Vitalfarbstoffe wird gezeigt, daß Phagozyten um Fremdsubstanzen bereits 10–50 Minuten nach ihrer Aufnahme in das Zytoplasma eine Hülle aus basophiler Substanz bilden. Es wird vermutet, daß saure Mukopolysaccharide die Träger der Basophilie sind. Die Beziehungen dieser vitalgefärbten, durch Phagozytose bzw. Pinozytose entstandenen Strukturen zu Vakuolen, die unter dem Einfluß basischer Farbstoffe selbst entstehen (Krinom), werden diskutiert.

In a comparative study cells grown in monolayer have been vitally stained with commercial dye samples and dyes purified by thin layer chromatography and column chromatography. The dyes used were derivatives of amino-substituted phenothiazine (thionine to methylene blue).

Fibroblasts grown in monolayer were subjected to vital staining by mepacrine (Atebrine®), neutral red and toludine blue under comparable conditions. These dyes induce the formation of autophagic vacuoles (autolysosomes) in the cytoplasm. The preservation of these structures, which are considered to be newly formed in the dimension of the light microscope, by seven different fixatives has been examined.

Es wird über das Verhalten von lebenden Blut- und Exsudatzellen nach Immersion in 2%–40% isotonischen Lösungen von ?-Globulin und Albumin berichtet.

This is the first of three studies on the relationship between Substantia granulofilamentosa and crinoma. Using the blood of phenylhydrazine-poisened guinea pigs, 57 cationic dyes of various chemical structures and selected basic substances of nondye character, are tested for their suitability to precipitate the substantia granulofilamentosa. Attention is paid to the various types of precipitates in order to find out whether they show individual morphologic features. Out of 57 dyes, 36—most of which are derivatives of heterocyclic anthracene — are suitable for the demonstration of the substantia granulofilamentosa. The group of dyes, which for the time ever have been used successfully for the demonstration of reticulocytes, includes: methylene green, toluylene blue, pseudo-isocyanine, neutral violet, and amethyst violet.

The personal dosemeter system of CERN based on the film badge must be able to evaluate the irradiation risks in the radiation environment around high energy accelerators. Over the past few years a critical re-appraisal of the detectors used in the light of the advent of new operational quantities in radiation protection has been made. Of special interest in this context is the comparison between the methods used at present for the evaluation of personal doses from hard photon irradiation and the quantities proposed on the one hand by ICRU and on the other hand by a group of experts in Switzerland. In the latter case the detector is exposed on a plexiglass block, a phantom which is preferred to a sphere for the calibration of dosemeters of a certain size. In addition, dose values expressed in this new quantity turn out to be numerically rather close to personal doses received in radiation fields of induced radioactivity and presently evaluated at CERN.

From following the development of personnel doses at CERN over the past six years it has become evident that work in areas of induced radioactivity is the principal cause of exposure. The results of photon dose measurements free-in-air and around a phantom are presented and discussed in the light of new quantities in individual monitoring. The importance of these results, with respect to the practical situation, is discussed and the problems of phantom size is mentioned. Finally, the results of dose measurements in the phantom are presented, since such information is important in cases where it becomes necessary to transform personnel doses into personal doses.