Peptidases of Trichobilharzia regenti (Schistosomatidae) and its molluscan host Radix peregra S. Lat. (Lymnaeidae): construction and screening of cDNA library from intramolluscan stages of the parasite

Trichobilharzia regenti is a neurotropic bird schistosome,causing cercarial dermatitis in humans. In this study, ZAP cDNA expression library from Radix peregra s. lat. hepatopancreases containing intramolluscan stages of T. regenti was constructed and screened using PCR with specific and degenerate primers, designed according to previously described serine and cysteine peptidases of other parasite species. Full-length sequences of cathepsins B1 and L, and two serine peptidases, named RpSP1 and RpSP2, were obtained.The protein-protein BLAST analysis and parallel control reactions with template from hepatopancreases of T. regenti non-infected snails revealed that only cathepsin B1 was of parasite origin. The remaining sequences were derived from the snail intermediate host, which implies that the initial source of parasite mRNA was contaminated by snail tissue.Regardless of this contamination, the cDNA library remains an excellent molecular tool for detection and identification of bioactive molecules in T. regenti cercariae.     

The impact of initial concentration of selected pharmaceuticals on their early stage of dissipation in soils

Journal of Soils and Sediments - Dissipation of pharmaceutical compounds entered into the natural environment is an important process minimizing the adverse effects on the living organisms. The aim...     

The use of RAPD technique for the identification and classification of Pisum sativum L. genotypes

Summary The genomic DNAs of 42 Pisum sativum genotypes, representing four wild and cultivated subspecies were used as templates in RAPD reactions. Amplification with eight decamer primers generated 149 polymorphic products. Genetic similarities of RAPD profiles were estimated via a coefficient of Jaccard and then the data were processed by cluster analysis (UPGMA). Each genotype was clearly identified and separated from the others. Our results show that RAPD technology is a rapid, precise and sensitive technique for identification of pea genotypes. However, the phylogenetic relationships within the Pisum sativum, which we tested by bootstrap analysis (Wagner parsimony), must be interpreted with caution.     

Evaluation of isoflavone aglycon and glycoside distribution in soy plants and soybeans by fast column high-performance liquid chromatography coupled with a diode-array detector

An ultrafast HPLC/UV-vis DAD method working at 254 nm was applied for the determination of isoflavone aglycons and glycosides (genistin, genistein, daidzein, daidzin, glycitin, glycitein, ononin, formononetin, sissotrin, and biochanin A) in roots, stems, leaves, and soy pods of soy plants and in soybeans of five varieties (Korada, Quito, Rita, OAC Erin, and OAC Vison). An Atlantis dC18 ultrafast RP chromatographic column (20 mm x 2.1 mm, 3 microm particle size) was applied for separation of the isoflavone aglycons and glycosides. A flow rate of the mobile phase (0.1% (v/v) acetic acid, pH 3.75-solvent A and methanol-solvent B) was 0.35 mL min(-1), and the column temperature was 36 degrees C. A linear gradient profile from 13 up to 22% B (v/v) from zero to 2.5 min, up to 30% B to 3.21 min, up to 35% B to 4 min, up to 40% B to 4.5 min, up to 50% B to 5.14 min, and followed by negative gradient up to 13% B to 7.71 min was used. The absolute limits of detection per sample injection (5 microL) were the highest for biochanin A (166.2 fmol) and the lowest for genistin (17.0 fmol), respectively. An accelerated solvent extraction (ASE) in combination with sonication was applied for isolation of biologically active compounds. A solid-phase extraction procedure was used to purify the extracts in the case of analysis of soy plants parts. The recoveries of 96-106% were obtained for the different concentrations of the isoflavone aglycons and glycosides and the different matrixes (overall RSDs 2-9%). The highest isoflavone concentrations were found in roots (12.5 microg g(-1) dry weight), while the amounts were about 3-1100 microg g(-1) fresh weight in different varieties of soybeans.     

Impacts of population growth, economic development, and technical change on global food production and consumption

Over the next decades mankind will demand more food from fewer land and water resources. This study quantifies the food production impacts of four alternative development scenarios from the Millennium Ecosystem Assessment and the Special Report on Emission Scenarios. Partially and jointly considered are land and water supply impacts from population growth, and technical change, as well as forest and agricultural commodity demand shifts from population growth and economic development. The income impacts on food demand are computed with dynamic elasticities. Simulations with a global, partial equilibrium model of the agricultural and forest sectors show that per capita food levels increase in all examined development scenarios with minor impacts on food prices. Global agricultural land increases by up to 14% between 2010 and 2030. Deforestation restrictions strongly impact the price of land and water resources but have little consequences for the global level of food production and food prices. While projected income changes have the highest partial impact on per capita food consumption levels, population growth leads to the highest increase in total food production. The impact of technical change is amplified or mitigated by adaptations of land management intensities.     

The V-antigen of Yersinia forms a distinct structure at the tip of injectisome needles

Many pathogenic bacteria use injectisomes to deliver effector proteins into host cells through type III secretion. Injectisomes consist of a basal body embedded in the bacterial membranes and a needle. In Yersinia, translocation of effectors requires the YopB and YopD proteins, which form a pore in the target cell membrane, and the LcrV protein, which assists the assembly of the pore. Here we report that LcrV forms a distinct structure at the tip of the needle, the tip complex. This unique localization of LcrV may explain its crucial role in the translocation process and its efficacy as the main protective antigen against plague.     

The needle length of bacterial injectisomes is determined by a molecular ruler

Size determination represents a fundamental requirement for multicomponent biological structures. Some pathogenic bacteria possess a weapon derived from the flagellum. Like the flagellum, this type-III secretion apparatus, called the injectisome, has a transmembrane basal body, but the external component is a needle-like structure instead of a hook and a filament. Here, we provide evidence that the length of this needle is determined by the size of a protein, YscP, acting as a molecular ruler.     

Transgenic RNAi reveals essential function for CTCF in H19 gene imprinting

The imprinted regulation of H19 and Insulin-like growth factor 2 expression involves binding of the vertebrate insulator protein, CCCTC binding factor (CTCF), to the maternally hypomethylated differentially methylated domain (DMD). How this hypomethylated state is maintained during oogenesis and the role of CTCF, if any, in this process are not understood. With the use of a transgenic RNA interference (RNAi)-based approach to generate oocytes with reduced amounts of CTCF protein, we found increased methylation of the H19 DMD and decreased developmental competence of CTCF-deficient oocytes. Our results suggest that CTCF protects the H19 DMD from de novo methylation during oocyte growth and is required for normal preimplantation development.     

The effects of chronic intermittent noise exposure on broiler chicken performance

Effects of different noise levels (70 or 80 dB) that broilers were exposed to during the entire fattening period and also the effect of the timing of the first exposure to intermittent noise in the course of fattening (day 1 vs. day 7) were monitored. After 7 days of exposure to intermittent noise, experimental chickens already showed a significant decrease in live body weight in comparison with the control group. The difference between the group exposed to intermittent noise at 70 dB and the group exposed to 80 dB levels was not statistically significant, although the mean live body weight of broilers in the latter was lower during the entire fattening period. The chickens exposed to intermittent noise from day 1 of age showed lower mean live weight throughout the fattening period compared to chickens exposed to the same level of intermittent noise only from day 7 of age, although at the end of fattening the difference was statistically significant only in chickens exposed to the higher level of intermittent noise (80 dB).     

Effect of terbutryn at environmental concentrations on early life stages of common carp (Cyprinus carpio L.)

The aim of this study was to assess the toxicity of terbutryn to early developmental stages of common carp (Cyprinus carpio). Based on accumulated mortality in the experimental groups, lethal concentrations of terbutryn were estimated at 36 day LC50 = 3.06 mg l⁻¹ terbutryn. Based on inhibition of growth in the experimental groups, lowest observed-effect concentration (LOEC) = 0.005 mg l⁻¹ terbutryn; and no observed-effect concentration (NOEC) = 0.0007 mg l⁻¹ terbutryn. Fulton’s condition factors were significantly lower in fish exposed to 2 mg l⁻¹ compared with controls. By day 30, fish exposed to 0.00002 mg l⁻¹ - real environmental concentration in Czech rivers - 0.02, 0.2, and 2 mg l⁻¹ terbutryn showed significantly lower mass and total length compared with controls. No significant negative effects on hatching or embryo viability were demonstrated at the concentrations tested, but significant differences in early ontogeny among groups were noted. Fish from the two highest tested concentrations (0.2 and 2 mg l⁻¹) showed a dose-related delay in development compared with the controls. At concentrations of 0.02, 0.2, and 2 mg l⁻¹ damage to caudal kidney tubules when compared to control fish was found.