Einfluß von Melatonin auf die Differenzierung der Osteoklasten mittels quantitativer Imageanalyse

Melatonin, das vor allem in der Zirbeldrüse synthetisiert wird, hat einen tageszeitlichen Rhythmus und wird hauptsächlich in der Nacht freigesetzt. Das Hormon steuert verschiedene physiologische und pathophysiologische Prozesse und unterstützt das Skelettwachstum und hemmt die Knochenresorption durch verminderte Expression von RANKL. Die derzeitigen Auswertesystemen verursachen verschiedene Probleme, die eine automatische Bestimmung der Osteoklasten nicht ermöglichen. Zurzeit werden die Osteoklasten mit TRAP Färbung sichtbar gemacht. Nachteil dieser Methode ist es, dass die gefärbten Zellen für weitere Analyse nicht verwendet werden können. Außerdem ist diese Färbung sehr fehleranfällig und zeitintensiv. Die Multinuklearität der Osteoklasten ist ein anderes Problem für die automatische Quantifizierung. Daher soll ein neues automatisches Imageanalysesystem etabliert werden um die Osteoklasten zu erkennen und um die Wirkungen von Arzneistoffe und Hormone auf diese Zellen zu untersuchen. Diese Arbeit analysierte die Effekte von Melatonin auf die Differenzierung der Osteoklasten. Die murinen Knochenmarkszellen und RAW 264.7 Zellen wurden mit verschiedenen Konzentrationen von Melatonin gemeinsam mit RANKL und M-CSF kultiviert. Bei 1µM haben sich Netzwerke gebildet, die mit dem automatischen Auswertesystem nicht bestimmt werden können. Bei Konzentrationen von 10 nM und 100 nM sind weniger Osteoklasten entstanden. Bei RAW 264.7 Zellen konnten mehr und größere Osteoklasten gezüchtet werden. Um die Osteoklasten von anderen Zellen zu unterscheiden, wurden spezifische Marker gefunden. Die verwendeten Antikörper waren gegen den Calcitoninrezeptor, α-Tubulin und Makrophagenantigen gerichtet. Jedoch hat der Antiköper gegen das Makrophagenantigen F4/80 nicht funktioniert. Im Ganzen kann man sagen, dass für die Quantifizierung von Osteoklasten ein automatisches Imageanalysesystem etabliert wurde, mit dem man die Wirkungen von Melatonin auf die Osteoklastogenese untersuchen konnte. Mithilfe dieses Systems sollen weitere Stoffe und Hormone, die das Osteoklastenwachstum beeinflußen, untersucht werden.Melatonin, which is synthesized primarily in the pineal gland, is released mainly at night. This hormone controls various physiological and pathophysiological processes and supports skeletal growth and inhibits bone resorption by decreased expression of RANKL. The current automatic image analysis systems cause is based on the cell recognition by single nuclei and, therefore, does not allow an automatic determination of multinucleated osteoclasts. Currently, osteoclasts are visualized by TRAP staining and their number is determined by manual counting. The disadvantage of this method is that the manual counting is time consuming and error-prone. Furthermore, stained cells can not be used for further analysis for expressed proteins. Therefore a new automatic image anlaysis system was generated, which recognizes osteoclasts based on cell membrane and cytoplasmic markers as well as on the number of nuclei. It should be suitable for the investigation of the effects of drugs and hormones on these cells. To evaluate this programm und to analyze the effects of melatonin on the differentiation of osteoclasts, the new image anlysis syste was tested. To distinguish the osteoclasts from other cells, specific markers were found. Antibodies were used directed against the calcitoninreceptor, α-tubulin and macrophage antigen. To test the system and evaluate the effect of melaton in on osteoclast foramtion, murine bone marrow cells and RAW 264.7 cells were cultured with various concentrations of melatonin together with RANKL and M-CSF. At concentrations of 10 nM and 100 nM Melatonin, the formation of osteoclasts was impaired. At 1 µM melatonin, networks of osteoclasts were formed, but these networks could not be determined with the automatic evaluation system. Cultivation of RAW 264.7 cells let to the formation of large multinucleated osteoclast-like cells, which could be counted be the automated image analysis system. Taken together, the data show that the automatic image analysis system for the quantification of osteoclasts is suitable to study the effects of hormones like melatonin on the formation of osteoclasts. Application of this system will allow ti investigate the effect of various substances and hormones on osteoclastogenesis and to determine their effect on the growth of multinucleated cells

Spatial Econometric Analysis of R&D Spillovers in Turkey

Kalayci, Elif/0000-0001-6658-0595WOS: 000372509600004Research and development (R&D) activities are of great significance in the long term development of firms and R&D expenditures have been studied by economists including Romer. This paper sets out to investigate the effects of R&D spillovers at provincial level with specific reference to Turkey. To serve this purpose, data published by Turkish Statistical Institute have been employed. The paper covers the period from 2003 to 2007. The number of data points is 342 subsequent to the aggregation of data at the firm level. Of the data aggregated, it has been found out that 162 of them are zero. While running the spatial econometric estimations, inverse distance and neighboring matrices were employed as the weighting matrix. The determinants of R&D activities employed in the study are size, represented by the number of employees; qualified labor, represented by the number of R&D employees; technology transfer represented by expenses for licensing; foreign ownership, Pavitt sector dummies, location dummies by the sea, border or airport. The results of the analyses suggest that when both weighting matrices are used, spatial lag and error yielded significant results. There is R&D knowledge spillovers at provincial level in Turkey, shown by the spatial spillover effects in nearly one third of the total effects

The diagnostic value of image guided percutaneous fine needle aspiration biopsy in equivocal mediastinal masses

Background and aims: The aim of this study was to assess the diagnostic value of image guided percutaneous fine needle aspiration (FNA) biopsy in equivocal mediastinal masses

SCLEROSING HEMANGIOMA OF THE LUNG CLINICOPATHOLOGIC AND IMMUNOHISTOCHEMICAL FEATURES

The aim of this study was to demonstrate the immunohistochemical profile of sclerosing hemangioma of the lung(SHL) as well as to analyse its clinicopathological features. Immunohistochemical analysis could be peroformed in 3 cases for antibodies against TTF-1, surfactant protein-B, NSE, Chromogranin A, synaptophsin, CD31, CD34, estrogen and progesteron receptors, cytokeratin 7 EMA, p53 and Ki-57. Positive immunoreactivity for EMA in both cells of SHL supports the epithelial origin of this tunor. Also, immunopositivity for TTF-1 (in both cells), cytokeratin 7 (in surface cells) and surfactant protein B (in surface cells) leads to pneumoctytic differentiation of tumor cells. Positivity for progersterone receptor antibody, although it was detected to be local, may be associated with the feamale predominancy of SHL. No evidence of a malignant phenotype was found by Ki-67 and p53 similar with literature. The results of our study were discussed and compared with literature

Association Between Cytomorphological and Clinicopathological Features of Nonsmall Cell Lung Carcinoma

The most important prognostic factor for lung carcinoma which is one of the most common cancer types, is the stage of the tumor. In this study, we assessed the prognostic importance of cytomorphological features of lung carcinoma in transthoracic fine needle aspiration biopsies with their clinicopathologic correlation

Plasma levels of interleukin-10 and nitric oxide in response to two different desflurane anesthesia flow rates

OBJECTIVE: This study investigated interleukin-10 and nitric oxide plasma levels following surgery to determine whether there is a correlation between these two variables and if different desflurane anesthesia flow rates influence nitric oxide and interleukin-10 concentrations in circulation. MATERIALS AND METHODS: Forty patients between 18 and 70 years and ASA I-II physical status who were scheduled to undergo thyroidectomy were enrolled in the study. INTERVENTIONS: Patients were allocated into two groups to receive two different desflurane anesthesia flow rates: high flow (Group HF) and low flow (Group LF). MEASUREMENTS: Blood samples were drawn at the beginning (t 0) and end (t 1) of the operation and after 24 h (t 2). Plasma interleukin-10 and nitric oxide levels were measured using an enzyme-linked-immunosorbent assay and a Griess reagents kit, respectively. Hemodynamic and respiratory parameters were assessed. RESULTS: There was no statistically significant difference between the two groups with regard to interleukin-10 levels at the times of measurement. Interleukin-10 levels were increased equally in both groups at times t 1 and t 2 compared with preoperative concentrations. For both groups, nitric oxide circulating concentrations were significantly reduced at times t 1 and t 2 compared with preoperative concentrations. However, the nitric oxide value was lower for Group HF compared to Group LF at t 2. No correlation was found between the IL-10 and nitric oxide levels. CONCLUSION: Clinical usage of two different flow anesthesia forms with desflurane may increase interleukin-10 levels both in Group HF and Group LF; nitric oxide levels circulating concentrations were significantly reduced at times t 1 and t 2 compared with preoperative concentrations; however, at 24 h postoperatively they were higher in Group LF compared to Group HF. No correlation was detected between interleukin-10 and nitric oxide levels