Identification and antifungal susceptibility of a large collection of yeast strains isolated in Tunisian hospitals

In this study, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used as a rapid method to identify yeasts isolated from patients in Tunisian hospitals. When identification could not be exstablished with this procedure, sequencing of the internal transcribed spacer with 5.8S ribosomal DNA (rDNA) (ITS1-5.8S-ITS2) and D1/D2 domain of large-subunit (LSU rDNA) were employed as a molecular approach for species differentiation. Candida albicans was the dominant species (43.37% of all cases), followed by C. glabrata (16.55%), C. parapsilosis (13.23%), C. tropicalis (11.34%), C. dubliniensis (4.96%), and other species more rarely encountered in human diseases such as C. krusei, C. metapsilosis, C. lusitaniae, C. kefyr, C. palmioleophila, C. guilliermondii, C. intermedia, C. orthopsilosis, and C. utilis. In addition, other yeast species were obtained including Saccharomyces cerevisiae, Debaryomyces hansenii (anamorph known as C. famata), Hanseniaspora opuntiae, Kodamaea ohmeri, Pichia caribbica (anamorph known as C. fermentati), Trichosporon spp. and finally a novel yeast species, C. tunisiensis. The in vitro antifungal activities of fluconazole and voriconazole were determined by the agar disk diffusion test and Etest, while the susceptibility to additional antifungal agents was determined with the Sensititre YeastOne system. Our results showed low incidence of azole resistance in C. albicans (0.54%), C. tropicalis (2.08%) and C. glabrata (4.28%). In addition, caspofungin was active against most isolates of the collection with the exception of two K. ohmeri isolates. This is the first report to describe caspofungin resistant isolates of this yeas

Diacerein retards cell growth of chondrosarcoma cells at the G2/M cell cycle checkpoint via cyclin B1/CDK1 and CDK2 downregulation

BACKGROUND: Chondrosarcoma is characterized for its lack of response to conventional cytotoxic chemotherapy, propensity for developing lung metastases, and low rates of survival. Research within the field of development and expansion of new treatment options for unresectable or metastatic diseases is of particular priority. Diacerein, a symptomatic slow acting drug in osteoarthritis (SYSADOA), implicates a therapeutic benefit for the treatment of chondrosarcoma by an antitumor activity. METHODS: After treatment with diacerein the growth behaviour of the cells was analyzed with the xCELLigence system and MTS assay. Cell cycle was examined using flow cytometric analysis, RT-PCR, and western blot analysis of specific checkpoint regulators. The status for phosophorylation of mitogen-activated protein kinases (MAPKs) was analyzed with a proteome profiler assay. In addition, the possible impact of diacerein on apoptosis was investigated using cleaved caspase 3 and Annexin V/PI flow cytometric analysis. RESULTS: Diacerein decreased the cell viability and the cell proliferation in two different chondrosarcoma cell lines in a dose dependent manner. Flow cytometric analysis showed a classical G2/M arrest. mRNA and protein analysis revealed that diacerein induced a down-regulation of the cyclin B1-CDK1 complex and a reduction in CDK2 expression. Furthermore, diacerein treatment increased the phosphorylation of p38α and p38β MAPKs, and Akt1, Akt2, and Akt 3 in SW-1353, whereas in Cal-78 the opposite effect has been demonstrated. These observations accordingly to our cell cycle flow cytometric analysis and protein expression data may explain the G2/M phase arrest. In addition, no apoptotic induction after diacerein treatment, neither in the Cal-78 nor in the SW-1353 cell line was observed. CONCLUSIONS: Our results demonstrate for the first time that the SYSADOA diacerein decreased the viability of human chondrosarcoma cells and induces G2/M cell cycle arrest by CDK1/cyclin B1 down-regulation

Alien Registration- Lohberger, Anton (Rockport, Knox County)

https://digitalmaine.com/alien_docs/13239/thumbnail.jp

MUG-Mel2, a novel highly pigmented and well characterized NRAS mutated human melanoma cell line

NRAS mutation in melanoma has been associated with aggressive tumor biology and poor prognosis. Although targeted therapy has been tested for NRAS mutated melanoma, response rates still appear much weaker, than in BRAF mutated melanoma. While plenty of cell lines exist, however, only few melanogenic cell lines retain their in vivo characteristics. In this work we present an intensively pigmented and well-characterized cell line derived from a highly aggressive NRAS mutated cutaneous melanoma, named MUG-Mel2. We present the clinical course, unique morphology, angiogenic properties, growth characteristics using in vivo experiments and 3D cell culture, and results of the exome gene sequencing of an intensively pigmented melanogenic cell line MUG-Mel2, derived from a cutaneous metastasis of an aggressive NRAS p. Q61R mutated melanoma. Amongst several genetic alterations, mutations in GRIN2A, CREBP, PIK3C2G, ATM, and ATR were present. These mutations, known to reinforce DNA repair problems in melanoma, might serve as potential treatment targets. The aggressive and fast growing behavior in animal models and the obtained phenotype in 3D culture reveal a perfect model for research in the field of NRAS mutated melanoma.Peer reviewe

Entwicklung und Optimierung nicht-kartesischer Techniken für die Magnetresonanzbildgebung

Die Magnetresonanzbildgebung von Proben mit kurzen effektiven Spin-Spin-Relaxationszeiten, z.B. der Lunge (T2* ca. 1 ms), erfordert kurze Echozeiten, die mittels Messsequenzen mit im k-Raum-Zentrum beginnenden Auslesetrajektorien erreicht werden. In dieser Arbeit wurden spiralförmige und radiale Ausleseschemata sowie zugehörige Fourier-Bildrekonstruktionsverfahren entwickelt und für die Bildgebung derartiger Proben an klinischen Tomographen bei einer Magnetfeldstärke von 1,5 T optimiert. Mit beiden Sequenztypen wurde eine auflösungsunabhängige minimale Echozeit von 0,5 ms erreicht. Typische Parameter einer schnellen spiralförmigen Abtastung resultieren in ca. 4- bis 8-fach größeren Auslesezeiten als bei der in dieser Hinsicht optimalen Radial-MRT, bei der Auslesezeiten von 1,3 bis 1,8 ms zu signifikant geringeren Signalverlusten und Blurring-Artefakten durch T2*-Relaxation führen. Beim Gridding-Bildrekonstruktionsalgorithmus liefert die Dichte-Präkompensation mit der besten der untersuchten Wichtungsfunktionen eine um 11% schmalere Halbwertsbreite (FWHM) als die Postkompensation, die dafür ein um 67% günstigeres Signal-Artefaktverhältnis bietet. In der in-vivo Anwendung konnte mittels der Radial-MRT erstmalig mit einer Gradientenechotechnik bei einer Messdauer von etwa 1 s/Schicht eine Darstellung feiner Gefäße der menschlichen Lunge mit einer effektiven Auflösung von unter 2 mm erreicht werden (nominelle Auflösung 1,4 mm). Im Lungenparenchym wurde T2* mit 0,6 bis 1,7 ms bestimmt. Durch die radiale parallele Bildgebung mit einem Reduktionsfaktor 4 bis 6 ist bei vergleichbarer Messzeit mit konventionellen Techniken eine deutliche Auflösungsverbesserung erzielbar