Visualization of a Ran-GTP gradient in interphase and mitotic Xenopus egg extracts

The small guanosine triphosphatase Ran is loaded with guanosine triphosphate (GTP) by the chromatin-bound guanine nucleotide exchange factor RCC1 and releases import cargoes in the nucleus during interphase. In mitosis, Ran-GTP promotes spindle assembly around chromosomes by locally discharging cargoes that regulate microtubule dynamics and organization. We used fluorescence resonance energy transfer-based biosensors to visualize gradients of Ran-GTP and liberated cargoes around chromosomes in mitotic Xenopus egg extracts. Both gradients were required to assemble and maintain spindle structure. During interphase, Ran-GTP was highly enriched in the nucleoplasm, and a steep concentration difference between nuclear and cytoplasmic Ran-GTP was established, providing evidence for a Ran-GTP gradient surrounding chromosomes throughout the cell cycle.     

Insect hormone activity of p-(1,5-dimethylhexyl)benzoic acid derivatives in Dysdercus species

Derivatives of p-(1,5-dimethylhexyl)benzoic acid are juvenile hormone analogs with selective action on the hemipteran insects of the family Pyrrhocoridae. Their juvenile hormone activity is constant on five species of Dysdercus; it is about ten times lower on Pyrrhocoris, and no activity has been detected on hemipterans of some other families. Absence of profound species-specific variations in the activity suggests that the most active compounds of this type can be used as selective pesticides against all species of Dysdercus.     

New reflected-light microscope for viewing unstained brain and ganglion cells

We have designed and constructed a new type of reflected-light microscope to form images including only light reflected near the plane of the object. This selectivity of image formation is based on a mechanical flying-spot technique. Objects difficult or impossible to see with earlier microscopes, such as unstained cells and cell processes in brains of living salamanders and in excised dorsal root ganglia of frogs, have been observed routinely with this microscope.     

Non-invasive measurement of fillet composition of four freshwater fish species by computer tomography

Thecrude fat and crude protein content of fillets was predicted by X-ray computertomography (CT) and determined by chemical analyses. Between 20 to 40 serialscans were taken of 48 freshwater fish and samples from the ground fillet withskin were prepared for chemical analyses. Fish species investigated were asfollows: common carp (n = 18), grass carp (n = 10), silver carp (n = 10) andpike-perch (n = 10), all collected directly from fishponds and processedimmediately. The variables of the prediction equations were taken from thedensity values of the Hounsfield scale, on a range between –90 and+150, by summing the frequencies within each interval of 10 values. Withprincipal component analysis fillet fat and crude protein content could beestimated with R² = 0.88 and R² = 0.87 accuracy,respectively. Three dimensional"volumetric" estimation of fat tissue was also carried out on the basis of fatindices developed at our lab. The fillet composition of the Cyprinid speciescould be evaluated well with the applied methods, but not that of pike-perchwhich had an extremely low fillet and body fat content.     

The content of total mercury and methylmercury in common carp from selected Czech ponds

The aim of the present study was to investigate concentrations of total mercury (THg) and methylmercury (MeHG) in the common carp from Czech ponds of Rožmberk, Spolsky, Nezmar, and Velky Bědny. Seven common carps (Cyprinus caprio) from each of the ponds were caught. Muscle tissue, the liver, and the soft and hard roe were used for the tests. Total mercury and methylmercury were found in all the muscle tissue samples examined. Detection limits for total mercury and methylmercury determination methods were 0.001 mg kg^–1 and 0.013 mg kg⁻¹ respectively. Methylmercury levels in the liver and gonads were below the method’s limits of detection. THg and MeHg concentrations in muscle tissues were 0.018–0.063 mg kg^–1 w.w. and 0.019–0.063 mg kg^–1 w.w. respectively. MeHg made up 90–100% of THg in muscle tissues. Total mercury and methylmercury concentrations were significantly (P < 0.05) higher in fish from the Spolsky pond than from the Nezmar pond. MeHg/THg ratios were significantly (P < 0.05) higher in fish from the Rožmberk, Spolsky and Nezmar ponds compared with fish from the Velky Bědny pond.     

Multivariate analysis of relationship between potato (Solanum tuberosum L.) yield,amount of applied elements,their concentrations in tubers and uptake in a long-term fertilizer experiment

The aim of this study was to investigate how potato yield, the concentrations of elements (N, P, K, Ca, Mg, S, As, Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn) in tubers and their uptake are affected by mineral N, P and K fertilizers, straw and pig slurry application.     

Prevalence, antibiotic resistance, virulence traits and genetic lineages of Staphylococcus aureus in healthy sheep in Tunisia

Nasal swabs of 163 healthy sheep were obtained from two farms and one abattoir in Tunisia during 2010. Samples were inoculated in Baird Parker agar and ORSAB medium for Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) recovery, respectively. MRSA was detected in 5 of these 163 samples (3%) in ORSAB medium, and one isolate per sample was further studied. MRSA isolates were mecA-positive, typed as ST153-CC80-t044-agrIII, and contained blaZ, ant(6)-Ia, aph(3')-IIIa, erm(C), tet(K), and fusB genes encoding penicillin, streptomycin, kanamycin, erythromycin, tetracycline and fusidic acid resistance, respectively. These MRSA isolates showed indistinguishable or closely related PFGE-patterns and harboured the lukF/lukS gene encoding the Panton-Valentine leukocidin and the luk-ED, hla, hld, and hlg(v) genes. Methicillin-susceptible S. aureus (MSSA) were recovered in 68 of the 163 samples (41.7%) and one isolate per sample was characterized. Most of MSSA (82.4%) showed susceptibility to the tested antibiotics with exceptions: penicillin (6%, carrying blaZ gene), tetracycline (19%, carrying tet(K) gene) and fusidic acid (9%). The following toxin-genes were identified among MSSA: tst (53 isolates), luk-M (52), luk-ED, hla, hlb, hld and hlg(v) (67), hlg (1), sec (49), sel (52), and the egc-cluster-like sen-sem-sei-seo-seg (1). Ten spa-types (two of them new ones) and nine sequence types (six new ones) were detected among the 73 S. aureus isolates, and they were ascribed to agr types I and III. All MRSA and MSSA isolates were able to coagulate bovine plasma and MRSA harboured the immune-evasion-gene-cluster type E. Conclusions. Nares of healthy sheep could be a reservoir of PVL-positive community-associated-MRSA and also of TSST-positive S. aureus isolates, with potential implications in public health.     

Effect of drought on yield variability of key crops in Czech Republic

The relationship between seasonal agricultural drought and detrended yields (within a period from 1961 to 2000) of selected crops was assessed in the conditions of the Czech Republic, which are to some extent representative of a wider area of Central Europe. Impact of water stress was analyzed using time series of yields for 8 crops (spring barley, winter wheat, grain maize, potato, winter rape, oats, winter rye and hay from permanent meadows) for 77 districts in the Czech Republic (average district area is 1025 km²). Relative version of Palmer’s Z-index (rZ-index or rZ-i) was used as a tool for quantification of agricultural drought. The monthly values of the rZ-index for each individual district were calculated as the spatial average (only for the grids of arable land). The study showed that severe droughts (e.g., in 1981 and 2000) are linked with significant reduction in yields of the main cereals and majority of other crops through the most drought prone regions. We found a statistically significant correlation (p ≤ 0.05) between the sum of the rZ-index for the main growing period of each crop and the yield departures of spring barley within 81% (winter wheat in 57%, maize in 48%, potato in 89%, oats in 79%, winter rye in 52%, rape in 39%, hay in 79%) of the analyzed districts. This study also defined the crop-specific thresholds under which a soil moisture deficit (expressed in terms of rZ-index) leads to severe impact at the district level. This can be expressed as the sum of the monthly rZ-index during the period of high crop sensitivity to drought; for spring barley it is ?5, winter wheat ?5, maize ?9, rape ?12, winter rye ?10, oat ?4, potato ?6 and for hay ?3. The length of the sensitive period is also crop-specific and includes the months that are important for the yield formation. The results show that yields of spring barley (and spring crops in general) are significantly more affected by seasonal water stress than yields of winter crops and hay from permanent meadows. The study proved that a severe drought spell during the sensitive period of vegetative season does have a quantifiable negative effect, even within more humid regions. These results demonstrate that, at least in some areas of the CR (and probably most of Central Europe), drought is one of the key causes of interannual yield variability.