Diapause of copepods as an element for stabilizing the parasite system of some fish helminths

The analysis of infection dynamics of copepods, which are the intermediate hosts of three helminth species of freshwater fishes (Triaenophorus crassus, Proteocephalus exiguus and P. percae), has shown that diapause in the life cycle of the copepods is favourable for preserving the infection in the waterbody until physiological prerequisites for successful infection of the final host are acquired. The peculiarity of a copepod's life cycle may determine the strategy of a parasite in its preimaginal phase as a waiting stage, and the duration of the residence of helminth larvae in copepods that have an obligatory diapause, is one of the elements the provide stability in parasite systems.     

High-efficiency gene transfer into cultured embryonic motoneurons using recombinant lentiviruses

Primary neurons are a common tool for investigating gene function for survival and morphological and functional differentiation. Gene transfer techniques play an important role in this context. However, the efficacy of conventional gene transfer techniques, in particular for primary motoneurons is low so that it is not possible to distinguish whether the observed effects are representative for all neurons or only for the small subpopulation that expresses the transfected cDNA. In order to develop techniques that allow high gene transfer rates, we have optimized lentiviral-based gene transfer for cultured motoneurons by using a replication-defective viral vector system. These techniques result in transduction efficacies higher than 50%, as judged by EGFP expression under the control of SFFV or CMV promoters. Under the same conditions, survival and morphology of the cultured motoneurons was not altered, at least not when virus titers did not exceed a multiplicity of infection of 100. Under the same cell culture conditions, electroporation resulted in less than 5% transfected motoneurons and reduced survival. Therefore we consider this lentivirus-based gene transfer protocol as a suitable tool to study the effects of gene transfer on motoneuron survival, differentiation and function.     

Trimethine cyanine dyes as fluorescent probes for amyloid fibrils: The effect of N,N′-substituents

The effect of various N,N′-substituents in the molecule of benzothiazole trimethine cyanine dye on its ability to sense the amyloid aggregates of protein was studied. The dyes are low fluorescent when free and in the presence of monomeric proteins, but their emission intensity sharply increases in complexes with aggregated insulin and lysozyme, with the fluorescence quantum yield reaching up to 0.42.     

The production of succinic acid by yeast Yarrowia lipolytica through a two-step process

The production of α-ketoglutaric acid by yeast Yarrowia lipolytica VKMY-2412 from ethanol and its subsequent chemical conversion to succinic acid (SA) were investigated. A highly effective and environmentally friendly process of α-ketoglutaric acid production was developed using a special pH-controlling strategy, in which the titration of the culture broth with KOH in the acid-formation phase was minimal, that allowed accumulation of only low amounts of inorganic wastes in the course of SA recovery. The culture broth filtrate containing α-ketoglutaric acid (88.7 g l^?1) was directly employed for SA production; the amount of SA produced comprised 71.7 g l^?1 with the yield of 70 % from ethanol consumed. SA was isolated from the culture broth filtrate in a crystalline form with the purity of 100 %. The yield of isolated SA was as high as 72 % of its amount in the culture broth filtrate. The antimicrobial and nematocidic effects of SA of microbial origin on pathogenic organisms that cause human and plant diseases were revealed for the first time.     

The Relation of Different-Scale Membrane Processes Under Nitric Oxide Influence

Rhythmic activity, synaptic transmission and propagation of excitation depend on the processes that occur at different cellular levels. Modulation of neuron activity results from the changes of plasma membrane and organelles properties. We aimed to study the relationship between changes of the ion channels activity, membrane microviscosity and amount of bound Ca²⁺ under the influence of nitric oxide (NO). We also investigated the effect of NO on the refractive index of neurons. We have shown that NO activates voltage-dependent K(K_V)-channels and leads to the decrease in the amount of membrane-bound and stored Ca²⁺. NO causes a decrease in the microviscosity of the membrane of cytosomes and altered refractive index of neurones. The latter can result from the modification of plasma membrane physico-chemical properties and structural changes in the cytoplasm (vesicles movement, reorganisation of cytoskeleton, etc.). It is suggested that long-term modifications of the physico-chemical properties of membranes and reorganisation in the cytoplasm affect neuron activity and signal transmission.     

A homolog of ScRAD5 is involved in DNA repair and homologous recombination in Arabidopsis

Rad5 is the key component in the Rad5-dependent error-free branch of postreplication repair in yeast (Saccharomyces cerevisiae). Rad5 is a member of the Snf2 ATPase/helicase family, possessing as a characteristic feature, a RING-finger domain embedded in the Snf2-helicase domain and a HIRAN domain. Yeast mutants are sensitive to DNA-damaging agents and reveal differences in homologous recombination. By sequence comparisons we were able to identify two homologs (AtRAD5a and AtRAD5b) in the Arabidopsis thaliana genome, sharing about 30% identity and 45% similarity to yeast Rad5. AtRad5a and AtRad5b have the same kind of domain organization with a higher degree of similarity to each other than to ScRad5. Surprisingly, both genes differ in function: whereas two independent mutants of Atrad5a are hypersensitive to the cross-linking agents mitomycin C and cis-platin and to a lesser extent to the methylating agent, methyl methane sulfonate, the Atrad5b mutants did not exhibit any sensitivity to all DNA-damaging agents tested. An Atrad5a/Atrad5b double mutant resembles the sensitivity phenotype of the Atrad5a single mutants. Moreover, in contrast to Atrad5b, the two Atrad5a mutants are deficient in homologous recombination after treatment with the double-strand break-inducing agent bleomycin. Our results suggest that the RAD5-dependent error-free branch of postreplication repair is conserved between yeast and plants, and that AtRad5a might be functionally homologous to ScRad5.     

BayMeth: improved DNA methylation quantification for affinity capture sequencing data using a flexible Bayesian approach

Affinity capture of DNA methylation combined with high-throughput sequencing strikes a good balance between the high cost of whole genome bisulfite sequencing and the low coverage of methylation arrays. We present BayMeth, an empirical Bayes approach that uses a fully methylated control sample to transform observed read counts into regional methylation levels. In our model, inefficient capture can readily be distinguished from low methylation levels. BayMeth improves on existing methods, allows explicit modeling of copy number variation, and offers computationally efficient analytical mean and variance estimators. BayMeth is available in the Repitools Bioconductor package.     

Near-to-perfect homeostasis: examples of universal aging rule which germline evades

Aging is considered to be a progressive decline in an organism's functioning over time and is almost universal throughout the living world. Currently, many different aging mechanisms have been reported at all levels of biological organization, with a variety of biochemical, metabolic, and genetic pathways involved. Some of these mechanisms are common across species, and others work different, but each of them is constitutive. This review describes the common characteristics of the aging processes, which are consistent changes over time that involve either the accumulation or depletion of particular system components. These accumulations and depletions may result from imperfect homeostasis, which is the incomplete compensation of a particular biological process with another process evolved to compensate it. In accordance with disposable-soma theory, this imperfection in homeostasis may originate as a function of cell differentiation as early as in yeasts. It may result either from antagonistic pleiotropy mechanisms, or be simply negligible as a subject of natural selection if an adverse effect of the accumulation phenotypically manifests in organism's post-reproductive age. If this phenomenon holds true for many different functions it would lead to the occurrence of a wide variety of aging mechanisms, some of which are common among species, while others unique, because aging is the inherent property of most biological processes that have not yet evolved to be perfectly in balance. Examples of imperfect homeostasis mechanisms of aging, the ways in which germ line escapes from them, and the possibilities of anti-aging treatment are discussed in this review.     

Differences in the processing of DNA ends in Arabidopsis thaliana and tobacco: possible implications for genome evolution

Surprising species-specific differences in non-homologous end-joining (NHEJ) of genomic double-strand breaks (DSBs) have been reported for the two dicotyledonous plants Arabidopsis thaliana and Nicotiana tabacum. In Arabidopsis deletions were, on average, larger than in tobacco and not associated with insertions. To establish the molecular basis of the phenomenon we analysed the fate of free DNA ends in both plant species by biolistic transformation of leaf tissue with linearized plasmid molecules. Southern blotting indicated that, irrespective of the nature of the ends (blunt, 5 or 3 overhangs), linearized full-length DNA molecules were, on average, more stable in tobacco than in Arabidopsis. The relative expression of a -glucuronidase gene encoded by the plasmid was similar in both plant species when the break was distant from the marker gene. However, if a DSB was introduced between the promoter and the open reading frame of the marker, transient expression was halved in Arabidopsisas compared to tobacco. These results indicate that free DNA ends are more stable in tobacco than in Arabidopsis, either due to lower DNA exonuclease activity or due to a better protection of DNA break ends or both. Exonucleolytic degradation of DNA ends might be a driving force in the evolution of genome size as the Arabidopsis genome is more than twenty times smaller than the tobacco genome.