Molecular characterization of atypical Chlamydia and evidence of their dissemination in different European and Asian chicken flocks by specific real-time PCR

Chlamydia psittaci is a zoonotic pathogen associated primarily with avian chlamydiosis. New chlamydial agents with suspected zoonotic potential were recently detected from domestic poultry in Germany and France indicating that the spectrum of Chlamydiaceae encountered in birds is not confined to a single chlamydial species. For further characterization, a specific real-time PCR targeting the conserved 16S rRNA gene was developed and validated for a specific detection of these atypical Chlamydiaceae. In order to address the epidemiological importance of the new chlamydial agents and their distribution, Chlamydiaceae-positive chicken samples collected from flocks from five different countries were examined. The results confirmed that C.psittaci is not the predominant chlamydial species among chickens examined and suggested that the new chlamydial agents could putatively be widespread in poultry flocks (France, Greece, Croatia, Slovenia and China at least) justifying their systematic investigation when poultry samples are submitted to laboratories for avian chlamydiosis diagnosis. Besides, 16S rRNA-based dendrogram, including sequences from both isolates of the new chlamydial agents or positive samples as well as representative sequences from species belonging to the order Chlamydiales, showed the new chlamydial agents to form a distinct line of descent separated from those of other chlamydial species, but clearly grouped within the family Chlamydiaceae. Finally, the phylogenetic tree inferred from the multi-locus sequence typing based on four housekeeping fragments (gatA, gidA, enoA and hflX) and the ompA-based dendrogram showed an almost identical topology of the new chlamydial agents with that recovered by 16S rRNA-based dendrogram. Interestingly, partial ompA gene sequences displayed considerable diversity among isolates.     

Densitometric analysis of dental implant placement between flapless technique and the two-stage technique--a pilot study

Flapless technique is a surgical approach of implant placement without raising a mucoperiosteal flap. Such approach has many advantages: shorter surgical treatment, minimal bleeding, postoperative discomfort for the patient is reduced; possibility of immediate loading of the inserted implant, faster procedure of implant placement and by that less time is needed for the complete implant-prosthetic restoration. Purpose of this pilot study was radiographic assessment of flapless technique and determination of its clinical values in comparison with two-stage dental implant technique through computerized densitometric analysis. The sample consisted of 10 patients with missing teeth in the premolar region in the upper jaw. An implant was placed in that position. In the first group of 5 patients the implants were inserted with the flapless technique, and in the other group of 5 patients implant insertion was done with a two-stage technique. All inserted implants were loaded with metal-ceramic crowns 3 months after placement. The patients were followed for 18 months through clinical follow-ups and radiovisiographical (RVG) images made after 3, 12 and 18 months. After comparing the average densities, the results showed similar decrease of density in both groups, conventional two-stage technique showed 3.24 and flapless technique 1.23. It can be concluded that flapless technique in everyday clinical usage has the same result as the two-stage dental implant technique.     

Explaining variability in the production of seed and allergenic pollen by invasive <Emphasis Type="Italic">Ambrosia artemisiifolia</Emphasis> across Europe

To better manage invasive populations, it is vital to understand the environmental drivers underlying spatial variation in demographic performance of invasive individuals and populations. The invasive common ragweed, Ambrosia artemisiifolia, has severe adverse effects on agriculture and human health, due to its vast production of seeds and allergenic pollen. Here, we identify the scale and nature of environmental factors driving individual performance of A. artemisiifolia, and assess their relative importance. We studied 39 populations across the European continent, covering different climatic and habitat conditions. We found that plant size is the most important determinant in variation of per-capita seed and pollen production. Using plant volume as a measure of individual performance, we found that the local environment (i.e. the site) is far more influential for plant volume (explaining 25% of all spatial variation) than geographic position (regional level; 8%) or the neighbouring vegetation (at the plot level; 4%). An overall model including environmental factors at all scales performed better (27%), including the weather (bigger plants in warm and wet conditions), soil type (smaller plants on soils with more sand), and highlighting the negative effects of altitude, neighbouring vegetation and bare soil. Pollen and seed densities varied more than 200-fold between sites, with highest estimates in Croatia, Romania and Hungary. Pollen densities were highest on arable fields, while highest seed densities were found along infrastructure, both significantly higher than on ruderal sites. We discuss implications of these findings for the spatial scale of management interventions against A. artemisiifolia.     

Effects of homocysteine and its related compounds on oxygen consumption of the rat heart tissue homogenate: the role of different gasotransmitters

Accumulating evidence indicates that microRNAs are implicated in tumor initiation and progression through negatively regulating oncogenes or tumor suppressor genes. In the present study, we report that the expression of miR-200a was significantly lower in renal cell carcinoma (RCC) specimens and RCC cell lines. Restoration of miR-200a suppressed cell growth, arrested cell cycle progression, and promoted cell apoptosis in RCC cell lines. We next used qRT-PCR array technology to identify Sirtuin 1 (SIRT1) as one of the downregulated proteins during miR-200a overexpression in 786-O cells. Following a further assay by luciferase reporter system, SIRT1 was validated as a direct target of miR-200a. Moreover, siRNA-mediated knockdown of SIRT1 could partially phenocopy the effects of miR-200a overexpression. In contrast, overexpression of truncated SIRT1 (without an endogenous 3′-UTR) could rescue the effect of miR-200a overexpression on 786-O cells, which suggested that SIRT1 3′-UTR is targeted by miR-200a specifically. These observations provide further evidence for a critical tumor-suppressive role of the miR-200a in RCC in addition to identifying a novel regulatory mechanism, which may contribute to SIRT1 upregulation in RCC.     

Naturally occurring compounds in differentiation based therapy of cancer

Differentiation of cancer cells entails the reversion of phenotype from malignant to the original. The conversion to cell type characteristic for another tissue is named transdifferentiation. Differentiation/transdifferentiation of malignant cells in high grade tumor mass could serve as a nonaggressive approach that potentially limits tumor progression and augments chemosensitivity. While this therapeutic strategy is already being used for treatment of hematological cancers, its feasibility for solid malignancies is still debated. We will presently discuss the natural compounds that show these properties, with focus on anthraquinones from Aloe vera, Senna, Rheum sp. and hop derived prenylflavonoids.     

Voltage Gated Potassium Channel Kv1.3 Is Upregulated on Activated Astrocytes in Experimental Autoimmune Encephalomyelitis

Kv1.3 is a voltage gated potassium channel that has been implicated in pathophysiology of multiple sclerosis (MS). In the present study we investigated temporal and cellular expression pattern of this channel in the lumbar part of spinal cords of animals with experimental autoimmune encephalomyelitis (EAE), animal model of MS. EAE was actively induced in female Dark Agouti rats. Expression of Kv1.3 was analyzed at different time points of disease progression, at the onset, peak and end of EAE. We here show that Kv1.3 increased by several folds at the peak of EAE at both gene and protein level. Double immunofluorescence analyses demonstrated localization of Kv1.3 on activated microglia, macrophages, and reactive astrocytes around inflammatory lesions. In vitro experiments showed that pharmacological block of Kv1.3 in activated astrocytes suppresses the expression of proinflammatory mediators, suggesting a role of this channel in inflammation. Our results support the hypothesis that Kv1.3 may be a therapeutic target of interest for MS and add astrocytes to the list of cells whose activation would be suppressed by inhibiting Kv1.3 in inflammatory conditions.     

Ontogenetic plasticity of anatomical and ecophysiological traits and their correlations in Iris pumila plants grown in contrasting light conditions

To better understand what directs and limits the evolution of phenotype, constraints in the realization of the optimal phenotype need to be addressed. That includes estimations of variability of adaptively important traits as well as their correlation structures, but also evaluation of how they are affected by relevant environmental conditions and development phases. The aims of this study were to analyze phenotypic plasticity, genetic variability and correlation structures of important Iris pumila leaf traits in different light environments and ontogenetic phases, and estimate its evolutionary potential. Stomatal density, specific leaf area, total chlorophyll concentration and chlorophyll a/b ratio were analyzed on I. pumila full‐sib families in the seedling phase and on the same plants after 3 years of growth in contrasting light conditions typical for ontogenetic stage in question. There was a significant phenotypic plasticity in both ontogenetic stages, but significant genetic variability was detected only for chlorophyll concentrations. Correlations of the same trait between different stages were weak due to changes in environmental conditions and difference in ontogenetic reaction norms of different genotypes. Ontogenetic variability of correlation structures was detected, where correlations and integration were higher in seedlings compared with adult plants 3 years later. Correlations were affected by environmental conditions, with integration being higher in the lower light conditions, but correlations between phases being stronger in the higher light treatment. These findings demonstrated that the analyzed traits can be selected and can mostly evolve independently in different environments and ontogenetic stages, with low genetic variability as a potentially main constraint.