Selection of suitable reference genes for gene expression studies in myxosporean (Myxozoa, Cnidaria) parasites

Myxozoans (Cnidaria: Myxozoa) are an extremely diversified group of endoparasites some of which are causative agents of serious diseases in fish. New methods involving gene expression studies have emerged over the last years to better understand and control myxozoan diseases. Quantitative RT-PCR is the most extensively used approach for gene expression studies. However, the accuracy of the results depends on the normalization of the data to reference genes. We studied the expression of eight commonly used reference genes, adenosylhomocysteinase (AHC1), beta actin (ACTB), eukaryotic translation elongation factor 2 (EF2), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), hypoxanthine-guanine phosphoribosyltransferase 1 (HPRT1), DNA-directed RNA polymerase II (RPB2), 18S ribosomal RNA (18S), 28S ribosomal RNA (28S) across different developmental stages of three myxozoan species, Sphaerospora molnari, Myxobolus cerebralis and Ceratonova shasta, representing the three major myxozoan linages from the largest class Myxosporea. The stable reference genes were identified using four algorithms: geNorm, NormFinder, Bestkeeper and ΔCq method. Additionally, we analyzed transcriptomic data from S. molnari proliferative and spore-forming stages to compare the relative amount of expressed transcripts with the most stable reference genes suggested by RT-qPCR. Our results revealed that GAPDH and EF2 are the most uniformly expressed genes across the different developmental stages of the studied myxozoan species.Fil: Kosakyan, Anush. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República ChecaFil: Alama Bermejo, Gema. Universidad Nacional del Comahue. Centro de Investigación Aplicada y Transferencia Tecnológica en Recursos Marinos "Almirante Storni". - Provincia de Río Negro. Ministerio de Agricultura, Ganadería y Pesca. Centro de Investigación Aplicada y Transferencia Tecnológica en Recursos Marinos "Almirante Storni". Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Centro Nacional Patagónico. Centro de Investigación Aplicada y Transferencia Tecnológica en Recursos Marinos "Almirante Storni"; Argentina. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República Checa. State University of Oregon; Estados UnidosFil: Bartošová-Sojková, Pavla. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República ChecaFil: Born-Torrijos, Ana. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República ChecaFil: Šíma, Radek. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República ChecaFil: Nenarokova, Anna. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República Checa. University of South Bohemia; República ChecaFil: Eszterbauer, Edit. Hungarian Academy Of Sciences; HungríaFil: Bartholomew, Jerri. State University of Oregon; Estados UnidosFil: Holzer, Astrid Sybylle. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República Chec

Software-Defined Optics in Last Mile for Research and Education in the Czech Republic

The Software Defined Networking (SDN) gained recognition due to its improvements at the packet switching and routing layers. The benefits of programmable, remotely controllable networking devices can be applied to the optical layer of contemporary networks as well. This work introduces CESNET's Czech Light~® family of devices and their role in enabling the SDN approach within the CESNET's production network. As a case study, an upgrade of the Cheb node in CESNET's network from a hard-spliced add-drop multiplexers to SDN-capable Reconfigurable Optical Add-Drop Multiplexers (ROADM) is presented. The upgrade improved operational capabilities of the network, including remote channel equalization, and the possibility to deploy new channels or lambdas without physical intervention. The deployment of ROADM also improved the optical properties of the network

The Ixodes ricinus salivary gland proteome during feeding and B. Afzelii infection: New avenues for an anti-tick vaccine

Introduction Borrelia burgdorferi sensu lato, the causative agents of Lyme borreliosis, are transmitted by Ixodes ticks. Tick saliva proteins are instrumental for survival of both the vector and spirochete and have been investigated as targets for vaccine targeting the vector. In Europe, the main vector for Lyme borreliosis is Ixodes ricinus, which predominantly transmits Borrelia afzelii. We here investigated the differential production of I. ricinus tick saliva proteins in response to feeding and B. afzelii infection. Method Label-free Quantitative Proteomics and Progenesis QI software was used to identify, compare, and select tick salivary gland proteins differentially produced during tick feeding and in response to B. afzelii infection. Tick saliva proteins were selected for validation, recombinantly expressed and used in both mouse and guinea pig vaccination and tick-challenge studies. Results We identified 870 I. ricinus proteins from which 68 were overrepresented upon 24-hours of feeding and B. afzelii infection. Selected tick proteins were successfully validated by confirming their expression at the RNA and native protein level in independent tick pools. When used in a recombinant vaccine formulation, these tick proteins significantly reduced the post-engorgement weights of I. ricinus nymphs in two experimental animal models. Despite the reduced ability of ticks to feed on vaccinated animals, we observed efficient transmission of B. afzelii to the murine host. Conclusion Using quantitative proteomics, we identified differential protein production in I. ricinus salivary glands in response to B. afzelii infection and different feeding conditions. These results provide novel insights into the process of I. ricinus feeding and B. afzelii transmission and revealed novel candidates for an anti-tick vaccine

anti-tick vaccines to prevent tick-borne diseases in Europe

Ixodes ricinus transmits bacterial, protozoal and viral pathogens, causing disease and forming an increasing health concern in Europe. ANTIDotE is an European Commission funded consortium of seven institutes, which aims to identify and characterize tick proteins involved in feeding and pathogen transmission. The knowledge gained will be used to develop and evaluate anti- tick vaccines that may prevent multiple human tick-borne diseases. Strategies encompassing anti-tick vaccines to prevent transmission of pathogens to humans, animals or wildlife will be developed with relevant stakeholders with the ultimate aim of reducing the incidence of tick-borne diseases in humans

Correlation of morphologic and moleculargenetic features of selected tumors

We present a series of 16 salivary gland tumors with histomorphological and immunohistochemical features reminiscent of secretory carcinoma of the breast. This is a hitherto undescribed and distinctive salivary gland neoplasm, with features resembling both salivary acinic cell carcinoma and low grade cystadenocarcinoma, as well as displaying strong similarities to breast secretory carcinoma. Microscopically, the tumors have a lobulated growth pattern and are composed of microcystic and glandular spaces with abundant eosinophilic homogenous or bubbly secretory material positive for PAS, mucicarmine, MUC1, MUC4 and mammaglobin. The neoplasms also show strong vimentin, S-100 protein, and STAT5a positivity. For this tumor we propose a designation mammary analogue secretory carcinoma of salivary glands (MASC). The 16 patients comprised 9 men and 7 women, with a mean age of 46 years (range 21-75). Thirteen cases occurred in the parotid gland, and one each in the minor salivary glands of the buccal mucosa, upper lip, and palate. The mean size of the tumors was 2.1 cm (range 0.7 to 5.5 cm). The duration of symptoms was recorded in 11 cases and ranged from 2 months to 30 years. Clinical follow-up was available in 13 cases, and ranged..

Quality of life in Roma Children in the Age between 11 and 15 Old Years in České Budějovice.

This work deals with the research quality of life of Roma aged 10-15 years in the České Budějovice. The theoretical section explains the concept of quality of life, it's development and the evaluation. It also describes the history of the Roma ethnic group from the beginning of the Roma migration from Europe to the modern history. This study further describes the Roma culture, the differences between female and male roles in their particle culture, analyzes of the Roma minority and their role in the European Union. The practical part of this study is focused on studying the physical, the mental, the social and the emotional status of the Roma respondents in the age group 10-15 years. This questionnaire survey confirmed the results of the National report on the health of the Roma population in the age group 10-15

Correlation of morphologic and moleculargenetic features of selected tumors

We present a series of 16 salivary gland tumors with histomorphological and immunohistochemical features reminiscent of secretory carcinoma of the breast. This is a hitherto undescribed and distinctive salivary gland neoplasm, with features resembling both salivary acinic cell carcinoma and low grade cystadenocarcinoma, as well as displaying strong similarities to breast secretory carcinoma. Microscopically, the tumors have a lobulated growth pattern and are composed of microcystic and glandular spaces with abundant eosinophilic homogenous or bubbly secretory material positive for PAS, mucicarmine, MUC1, MUC4 and mammaglobin. The neoplasms also show strong vimentin, S-100 protein, and STAT5a positivity. For this tumor we propose a designation mammary analogue secretory carcinoma of salivary glands (MASC). The 16 patients comprised 9 men and 7 women, with a mean age of 46 years (range 21-75). Thirteen cases occurred in the parotid gland, and one each in the minor salivary glands of the buccal mucosa, upper lip, and palate. The mean size of the tumors was 2.1 cm (range 0.7 to 5.5 cm). The duration of symptoms was recorded in 11 cases and ranged from 2 months to 30 years. Clinical follow-up was available in 13 cases, and ranged...1 Univerzita Karlova v Praze, Lékařská fakulta v Plzni Šiklův patologicko anatomický ústav Korelace morfologických a molekulárně genetických vlastností vybraných nádorů Radek Šíma Autoreferát doktorské dizertační práce Plzeň 2010 2 Dizertační práce byla zpracována v rámci kombinované formy doktorského studijního programu na Šiklově patologicko-anatomickém ústavu LF UK v Plzni v letech 2006-2010. Uchazeč: Radek Šíma Šiklův patologicko anatomický ústav LF UK v Plzni Školitel: Prof. MUDr. Alena Skálová, CSc. Šiklův patologicko anatomický ústav LF UK v Plzni Oponenti: Doc. MUDr. Ludmila Boudová, PhD. Šiklův patologicko anatomický ústav LF UK v Plzni Prof. MUDr. Jiří Ehrmann, PhD. Ústav histologie a embryologie LF UP v Olomouci Stanovisko k dizertační práci vypracoval vedoucí školícího pracoviště: Prof. MUDr. Michal Michal, Šiklův patologicko anatomický ústav LF UK v Plzni Autoreferát byl rozeslán dne: Obhajoba se koná na Šiklově patologicko-anatomickém ústavu, Dr. E. Beneše 13, Plzeň, před komisí pro obhajoby doktorských dizertačních prací v oboru patologie dne 27.4.2010 ve 14:00. S dizertační prací je možné se...Šiklův ústav patologieLékařská fakulta v PlzniFaculty of Medicine in Pilse

FFF 3D printing in electronic applications: Dielectric and thermal properties of selected polymers

The present study is a focused and comprehensive analysis of the dielectric and thermal properties of twenty-four 3D printed polymers suitable for fused filament fabrication (FFF) in electronic applications. The selected polymers include various thermoplastic elastomers, such as thermoplastics based on polycarbonate (PC), polyethylene terephthalate glycol (PETG), and acrylonitrile butadiene styrene (ABS-T). Their overall thermal behavior, including oxidation stability, glass transition, and melting temperature, was explored using simultaneous thermal analysis (STA) and differential scanning calorimetry (DSC). Considering their intended usage in electronic applications, the dielectric strength (Ep) and surface/volume resistivity (rs/rv) were comprehensively tested according to IEC 60243-1 and IEC 62631-3, respectively. The values of the dielectric constant and loss factor were also determined by broadband dielectric spectroscopy (BDS). While, on the one hand, exceptional dielectric properties were observed for some thermoplastic elastomers, the materials based on PCs, on the other hand, stood out from the others due to their high oxidation stability and above average dielectric properties. The low-cost materials based on PETG or ABS T did not achieve thermal properties similar to those of the other tested polymers; nevertheless, considering the very reasonable price of these polymers, the obtained dielectric properties are promising for undemanding electronic applications