Mycoplasma and host interaction: in vitro gene expression modulation in Mycoplasma synoviae and infected chicken chondrocytes

The complex interplay between Mycoplasma synoviae and chicken chondrocytes (CCH), which come into direct contact during infectious synovitis, has been examined at the level of gene expression. Our previous studies demonstrated a significant influence of M. synoviae on the level of CCH gene expression. Here, we show for the first time that in vitro co-cultivation of M. synoviae and CCH also induces upregulation of gene expression in this mycoplasma. We observed significantly increased expression of genes important for M. synoviae pathogenicity, including cysteine protease cysP, neuraminidase nanH, haemagglutinin vlhA, and the putative nuclease MS53_0284. Moreover, the pattern of gene expression was dependent on the infection environment. In CCH, significant changes in the expression of genes encoding catabolic enzymes of the cartilage extracellular matrix (cathepsins B, K and L, aggrecanase ADAM10, and matrix metalloproteinase MMP2) were demonstrated. Infection of CCH with M. synoviae also elevated the expression of the gene encoding peptidyl arginine deiminase, type III (PADI3), which is responsible for the post-translational citrullination of proteins

Production, Characterization and Use of Monoclonal Antibodies Recognizing IgY Epitopes Shared by Chicken, Turkey, Pheasant, Peafowl and Sparrow

Chicken antibodies are not only a part of immune defense but are more and more popular commercial products in form of chicken polyclonal, monoclonal or recombinant antibodies. We produced and characterized mouse monoclonal antibodies (mAbs) that recognize epitopes located on heavy or light chain of chicken immunoglobulin Y (chIgY) shared also by some other Phasianidae birds. The use of mAbs 1F5 and 2F10 that recognize heavy chain on chIgY common epitopes was demonstrated on immunoglobulins of turkey, pheasant and peafowl. Chicken IgY light chain specific mAb 3E10 revealed the presence of common epitopes on immunoglobulins of turkey, pheasant and sparrow. Monoclonal antibody clone 1F5/3G2 was used to prepare horseradish peroxidase (HRP) conjugate and immunoadsorbent column. Conjugated mAbs were demonstrated to be excellent secondary antibodies for diagnostics of certain infections in different avian species. Since they do not react with mammalian immunoglobulins using our mAbs as secondary antibodies in human serodiagnostics would minimize background staining that appears when using mouse detection system. In dot immunobinding assay (DIBA) and immunoblot assay they recognized specific IgY antibodies against Mycoplasma synoviae, Mycoplasma gallisepticum and Newcastle disease virus in sera of infected or vaccinated birds. Immunoadsorption as a method for removal of IgY from samples in which Mycoplasma synoviae specific IgY was predominant immunoglobulin class enabled more exact demonstration of specific IgA and IgM antibodies. Herein we are presenting effective mAbs useful in diagnostics of avian and mammalian infections as well as in final steps of detection and purification of chicken antibodies and their subunits produced in vivo or in vitro as polyclonal, monoclonal or recombinant antibodies

TONE KRALJ AND HIS CHURCH PAINTINGS IN THE SLOVENIAN LITTORAL AREA

V diplomski nalogi želim predstaviti Toneta Kralj in njegove cerkvene poslikave na slovenskem Primorskem. Tone Kralj je od začetka dvajsetih do poznih šestdesetih let na Primorskem ustvaril gigantski likovni opus, s katerim je skušal utrjevati slovensko nacionalno zavest na območju, ki je po letih 1918 pripadalo Italiji. Poslikal je več kot štirideset cerkva. Doslej še ni bil narejen sistematičen popis vseh Kraljevih poslikav na Primorskem, česar se loteva pričujoča diplomska naloga. Za okvir topografskega popisa si jemlje slovenske državne meje, v interpretaciji pa skuša naloga odpreti vprašanje stilnega razmerja obravnavanih del. Na začetku naloge na kratko predstavim Kraljevo umetniško pot, začetek cerkvenega slikarstva in takratno zgodovinsko dogajanje. V nadaljevanju obravnavana dela razdelim na tri poglavja glede na slikarjev stilni razvoj. Namen naloge je narediti sistematičen popis poslikav na slovenskem Primorskem in jih interpretirati glede na ikonografska izhodišča.Tone Kralj and his church paintings in the Slovenian Littoral are presented in the diploma. From the early 20s to the late 60s Tone Kralj created a gigantic atristic opus in the Littoral region with which he tried to strengthen the Slovenian national consciousness in the area which has belonged to Italy from 1918. He has painted more than fourty churches. Till now no systematic list of his paintings in the Littoral region has been made and is thus undertaken in this diploma. Slovenian national borders represent a topographical frame and the question of stylistic proportion of the discussed works is dealt with in the interpretation. At the beginning of this paper Karlj\u27s atrictic path, the beginning of church painting and the hystorical action at that time are presented. Further, I devide the discussed works in three chapters according to the painter\u27s stylistic development. The aim of the paper is to create a systematic list of paintings in Slovenian Littoral and to interpret them accodring to their iconographic origin

The Impact of HRM Practices on the Performance of Municipalities. The Case of Slovenia

On the basis of existing research, the paper presents a model for analyzing the impact of the use of human resource management practices on organizational performance. The model allows the assessment of these impacts after analyzing the use of human resource management practices and the achieved performance. The model includes a survey questionnaire on human resource management practices, a principal component analysis of the results of the questionnaire, a DEA model to determine the performance and a linear regression model of the impact of human resource management practices on the performance of Slovenian municipalities.The results of the research confirm a negative (focus on quality and job security) and positive (supervisory practices) impact of the use of certain human resource management practices in Slovenian municipalities on their organizational performance. The findings indicate the need for balancing between focus on quality, employee security, and supervisory practices for better individual and organizational performance in Slovenian local administration.</p

Proteolytically activated CRAC effectors through designed intramolecular inhibition

Highly regulated intracellular calcium entry affects numerous cellular physiological events. External regulation of intracellular calcium signaling presents a great opportunity for the artificial regulation of cellular activity. Calcium entry can be mediated by STIM proteins interacting with Orai calcium channelstherefore, the STIM1−Orai1 pair has become a tool for artificially modulating calcium entry. We report on an innovative genetically engineered protease-activated Orai activator called PACE. CAD self-dimerization and activation were inhibited with a coiled-coil forming peptide pair linked to CAD via a protease cleavage site. PACE generated sustained calcium entry after its activation with a reconstituted split protease. We also generated PACE, whose transcriptional activation of NFAT was triggered by PPV or TEV protease. Using PACE, we successfully activated the native NFAT signaling pathway and the production of cytokines in a T-cell line. PACE represents a useful tool for generating sustained calcium entry to initiate calcium-dependent protein translation. PACE provides a promising template for the construction of links between various protease activation pathways and calcium signaling

Vanadate from air pollutant inhibits hrs-dependent endosome fusion and augments responsiveness to toll-like receptors.

There is a well-established association between exposure to air pollutants and pulmonary injuries. For example, metals found in ROFA (residual oil fly ash) increase susceptibility of mice as well as humans to microbial infections. In our research, we have found that vanadate substantially increased the response of several Toll-like receptors (TLRs) to stimulation with their ligands. Although vanadate caused generation of reactive oxygen species (ROS), the addition of ROS scavenger N-acetyl cysteine (NAC) had no effect on augmented lipopolysaccharide (LPS) stimulation. We further showed that vanadate inhibits endosome fusion. This effect was determined by measuring the size of endosomes, NF-κB activity and TLR4 degradation in Hrs (hepatocyte growth factor-regulated tyrosine kinase substrate) overexpressed cells. Moreover, we identified the role of Hrs phosphorylation in these processes. Based on our findings, we can conclude that vanadate potentiates TLR4 activity by increasing Hrs phosphorylation status, reducing the size of Hrs/TLR4-positive endosomes and impacting TLR4 degradation, thus contributing to the detrimental effects of air pollutants on human health