BACKGROUND There is great interest in circulating microRNAs (miRNAs) as disease biomarkers. Translating promising miRNAs into validated clinical tests requires the characterization of many preanalytical and analytical parameters. METHODS miRNAs were extracted from serum and plasma samples of healthy volunteers, and miRNAs known to be present in serum and plasma (miR-15b, miR-16, miR-24, and miR-122) were amplified by reverse-transcription quantitative PCR. Stability and the effects of hemolysis were determined. Assay variation and its components, including the effect of adding control miRNA, were assessed by nested ANOVA. RESULTS miRNA concentrations were higher in plasma than in serum. Processing of plasma to remove subcellular/cellular components reduced miRNA concentrations to those of serum. The miRNAs analyzed were stable refrigerated or frozen for up to 72 h and were stable at room temperature for 24 h. Hemolysis increased the apparent concentration of 3 of the miRNAs. The total variability of replicate miRNA concentrations was <2.0-fold, with most of the variability attributable to the extraction process and interassay imprecision. Normalizing results to those of spiked exogenous control miRNAs did not improve this variability. CONCLUSIONS Detailed validation of the preanalytical steps affecting miRNA detection and quantification is critical when considering the use of individual miRNAs as clinical biomarkers. Unless these causes of imprecision are considered and mitigated, only miRNAs that are extremely up- or downregulated will be suitable as clinical biomarkers.

The immunosuppressive properties of morbilliviruses including measles and canine distemper virus (CDV) are well known, but the host cells supporting infection are poorly characterized. To identify these cells, a recombinant CDV expressing green fluorescent protein was produced by reverse genetics based on a wild-type strain lethal for ferrets. This recombinant virus fully retained virulence and blazed three lymphocyte-based pathways through the immune system of its host: first, it infected rapidly and massively circulating B and T cells; second, it took over and damaged secondary lymphatic organs including spleen, lymph nodes, and gut-associated and mucosal lymphoid tissues; third, it infected most thymocytes. In contrast, replication in epithelial cells was initially not detectable, but substantial before host death. Thus, CDV initially infects lymphocytes and massively replicates therein, thereby causing immunosuppression and preparing systemic invasion and host escape.virus ͉ measles ͉ immunosuppression

Fish introductions, particularly in areas of high biological diversity and endemism, represent a major threat for biodiversity. In the Balkan Peninsula, 60 fish species have been introduced to date, of which 36 have become naturalized in inland waters. Since the Balkans are one of the world's 35 biodiversity hot spots, this large presence of alien fish species poses a serious threat for the stability of freshwater ecosystems and the survival of the native ichthyofauna and of aquatic biodiversity in general. The motivation for the introductions, and the historical timeline, varies among the Balkan states. Despite recent attempts to implement and align legislation aimed at preventing the introduction of potentially invasive species, and the implementation of rigorous controls of introductions and increased protection of open waters, the majority of current introductions remain intentional, primarily via aquaculture. This review article provides a historical overview of freshwater fish introductions, the motivation behind them and the current distribution of alien freshwater fishes in the Balkans. The ecological implications and future perspectives concerning alien fish species in the region are also discussed.

Seed germination is first critical and the most sensitive stage in the life cycle of plants compromise the seedlings establishment. Salt and drought tolerance testing in initial stages of plant development is of vital importance, because the seed with more rapid germination under salt or water deficit conditions may be expected to achieve a rapid seedling establishment, resulting in higher yields. The aim of this study was to determine whether the pea seed germination and seedling growth were inhibited by the salt toxicity and osmotic effect during the seedling development, and also identification of the sensitive seedling growth parameters in response to those stresses. Based on the obtained results, pea has been presented to be more tolerant to salt than water stress during germination and early embryo growth. Investigated cultivars showed greater susceptibility to both abiotic stresses when it comes growth parameters compared to seed germination.

SUMMARyTomatoes are parasitized by a number of pathogens, including Fusarium oxysporum f. sp. lycopersici, causal agent of fusarium wilt. Fresh vegetable fruits can be contaminated with various fungi that produce mycotoxins, which is an important issue for human health. The objective of this paper was to isolate, determine, and identify causal organisms of tomato wilt and fruit rot, based on the pathogens morphological and molecular characteristics. Samples of diseased plants showing symptoms of tomato wilt were collected from different localities in the production region of Vojvodina. Fruits with symptoms of fusarium rot were collected from storage and warehouses. The isolation and morphological determination of the fungus were performed on PDA and Czapek's nutrient media. Isolates from diseased plants growing in field, designated as TFW1-TFW12 and seven isolates from diseased tomato fruits (TFM1-TFM7) were chosen for further investigation. For identification of the fungal isolates, the polymerase chain reaction (PCR) was also used. The EF1/EF2 primer pair was used for molecular identification of Fusarium sp. Nine analyzed samples were found to contain DNA fragments 700 bp in size.