Structural changes of eIF4E upon binding to the mRNA 5' monomethylguanosine and trimethylguanosine Cap

Recognition of the 5' cap by the eukaryotic initiation factor 4E (eIF4E) is the rate-limiting step in the ribosome recruitment to mRNAs. The regular cap consists of 7-monomethylguanosine (MMG) linked by a 5'-5' triphosphate bridge to the first transcribed nucleoside, while some primitive eukaryotes possess a N (2), N (2),7-trimethylguanosine (TMG) cap structure as a result of trans splicing. Mammalian eIF4E is highly specific to the MMG form of the cap in terms of association constants and thermodynamic driving force. We have investigated conformational changes of eIF4E induced by interaction with two cap analogues, 7-methyl-GTP and N (2), N (2),7-trimethyl-GTP. Hydrogen-deuterium exchange and electrospray mass spectrometry were applied to probe local dynamics of murine eIF4E in the apo and cap-bound forms. The data show that the cap binding induces long-range conformational changes in the protein, not only in the cap-binding pocket but also in a distant region of the 4E-BP/eIF4G binding site. Formation of the complex with 7-methyl-GTP makes the eIF4E structure more compact, while binding of N (2), N (2),7-trimethyl-GTP leads to higher solvent accessibility of the protein backbone in comparison with the apo form. The results suggest that the additional double methylation at the N (2)-amino group of the cap causes sterical effects upon binding to mammalian eIF4E which influence the overall solution dynamics of the protein, thus precluding formation of a tight complex.     

Alternations in phenolic acids content in developing rye grains in normal environment and during enforced dehydration

A series of high pressure liquid chroamtography analyses revealed the presence of five phenolic acids in rye caryopses (vanillic, caffeic, p-coumaric, ferulic and sinapic), three of which (p-coumaric, ferulic and sinapic) were found in the free phenolic fraction. Ferulic acid was predominant, both among free acids and total phenolic acids (i.e. free, liberated from soluble esters and glycosides). The highest content of the free phenolic acids in rye caryopses was observed at the beginning of development, when on 22 DAF it was estimated at 11.55 μg·g⁻¹ DW. During dehydratation the total level of free phenolic acids in rye caryopses decreased in all investigated samples. Although total phenolic acids contents in all samples of unripe rye caryopses always decreased after dehydration, in rye sample collected in full ripeness (57 DAF), the amount of these compounds increased after the enforced dehydration. It should be added that in ester-bound-soluble phenolic acids fraction (the largest part in the total phenolic acids fraction), irrespective of the total amount decrease, much increase of sinapic acid content in this fraction was observed after dehydratation treatment in all investigated samples of caryopses of various ripeness. During the development and ripening of rye caryopses, a gradual increase in the precocious germination ability of the grain was observed. The enforced dehydration stimulated the process of precocious germination of developing and ripening rye caryopses. A possible role of phenolics in preventing precocious germination and acclimation to dehydration of developing and ripening rye grains is discussed.     

Trophic state of a lowland reservoir during 10 years after restoration

The restored Maltañski Reservoir was studied from its filling with water in 1990 till 2000. Total phosphorus, chlorophyll-a, and Secchi depth, as well as the Carlson's trophic state index (TSI) values based on those three parameters showed characteristic patterns of changes among seasons and years. Within each year, the lowest trophic state was usually observed in winter and the highest in summer. Because of the high loads of phosphorus received by the reservoir, this element did not limit primary production. TSI values calculated on the basis of total phosphorus were always markedly higher than calculated on chlorophyll-a and Secchi depth (similar to each other). The trophic upsurge phase lasted only a few months after the filling of the reservoir in 1990. Similar symptoms were observed after its refilling in the spring of 1993. The trophic depression phase lasted until the end of 1995. After that time a significant correlation between phosphorus concentration in the reservoir and in river waters flowing into the reservoir was observed. The successive phases of reservoir ageing, determined on the basis of phosphorus concentration, were not accompanied by changes in chlorophyll-a content. The influence of the top-down mechanism (biomanipulation effect) resulted in relatively low values of chlorophyll-a after the filling of the reservoir with water in 1990 and in 1993. As early as 1992 chlorophyll-a values reached a very high level and stayed at that level until the end of the study in 2000 (except for the short decline in 1993).

     

X-ray fluorescence determination of the loss of chosen electrolytes in the urine of children with a congenital cyanotic heart defect and after heart transplantation

In this paper results of the analysis of urine samples of healthy children and children with a congenital cyanotic heart defect and after heart transplantation are presented. The analysis of urine samples was carried out by X-ray fluorescence spectrometry with wavelength dispersion and using CRM urine Seronorm as a reference. It was found that for patients with a congenital cyanotic heart defect the loss of electrolytes like Na, Cl and K was increased. Moreover, urine samples of children from areas of different degree of environmental pollution were analysed. We observed (as expected) higher concentrations of heavy metals in the urine of children from ecological polluted areas.     

Cumulative consumption of the lake macrophyte Elodea by abundant generalist invertebrate herbivores

Previous investigations have shown that macrophyte biomass can be substantially reduced by invertebrate herbivores but have not provided evidence for the links between the magnitude of the observed damage and the densities of herbivores. The results of this study support the hypothesis that the abundant occurrence of the epiphytic generalist herbivores may result in their cumulative consumption which, in turn, can be regarded as the mechanism responsible for often observed relatively high level of herbivory on freshwater macrophytes. The percentage of Elodea sp. biomass consumed by invertebrates was estimated for six European lakes, based on analysis of gut contents, daily rations and the density of epiphytic herbivores. Although the daily ration of these invertebrates when feeding upon Elodea averaged only 14.6% of their dry mass, their biomass was relatively high (from 0.163 to 1.161 g DW per 100 g DW plant). The estimated percentage of Elodea biomass consumed during one summer month by epiphytic invertebrates ranged from 0.5 to 5.9%. These values, after extrapolating to the whole growing season would mean that the biomass of Elodea lost to herbivory was between about 2 and 23%, an estimate which are within the range of consumption reported by other authors.     

Aeromonas spp.-mediated cell-contact cytotoxicity is associated with the presence of type III secretion system

In the study we examined the production of cytotonic and cytotoxic toxins and the presence of a type III secretion system (TTSS) in 64 Aeromonas spp. strains isolated from fecal specimens of patients with gastroenteritis. We observed that contact of the bacteria with host epithelial cells is a prerequisite for their cytotoxicity at 3 h incubation. Cell-contact cytotoxic activity of the strains was strongly associated with the presence of the TTSS. Culture supernatants of the strains induced low cytotoxicity effects at the same time of incubation. Cell-free supernatants of 61 (95%) isolates expressed cytotoxic activity which caused the destruction of HEp-2 cells at 24 h. Moreover, 44% strains were cytotonic towards CHO cells and 46% of strains invaded epithelial cells.     

Ornithine transcarbamylase (OTC) deficiency in a female patient with a de novo deletion of the paternal X chromosome

Summary A girl with ornithine transcarbamylase (OTC) deficiency was investigated for molecular and cytogenetic abnormalities that might explain this phenotype. Analysis with polymorphic DNA markers indicated that the patient did not inherit paternal alleles of the OTC locus, but that she did inherit the proximal locus DXS7 and the long arm of chromosome X. High-resolution cytogenetic analysis of the patient indicated a deletion of Xp11.4-p21, whereas both parents had normal karytoypes. Since the mother might be heterozygous according to biochemical tests, a second mutation within the maternal OTC gene cannot be excluded.     

The effect of industrial pollution on copper,lead, zinc and cadmium concentration in xylem rings of resistant (Populus marilandica) and sensitive (P. balsamifera) species of poplar

Summary The patterns of radial distribution of copper, lead, zinc and cadmium in two species of poplar, growing under polluted conditions near a copper smelter, were investigated. Populus marilandica is very resistant to such pollution, but P. balsamifera is sensitive. Soils were examined for concentrations of these metals, and only a surface layer of soil was polluted with copper and lead. P. balsamifera absorbed much more cadmium than P. marilandica under the same conditions. P. marilandica shows a pattern of copper radial distribution with significantly increased concentration of the meral in the layers in the middle of the trunk compared to P. balsamifera. Zinc and cadmium have a pattern with increased metal concentration from the bark towards the pith of the tree. In the case of copper and lead the pattern indicates increased concentrations of these metals in the rings adjacent to the pith while the pattern in other parts of the tree remains unconfirmed due to poor precision. Poplar xylem seems to accumulate zinc and cadmium while the content of lead in xylem is much lower than in the adjacent soil.This paper is dedicated to the memory of Professor Stefan Bialobok, Institute of Dendrology, Polish Academy of Sciences in Kornik     

Endogenously generated DNA nucleobase modifications source,and significance as possible biomarkers of malignant transformation risk,and role in anticancer therapy

The DNA of all living cells undergoes continuous structural and chemical alteration, which may be derived from exogenous sources, or endogenous, metabolic pathways, such as cellular respiration, replication and DNA demethylation. It has been estimated that approximately 70,000 DNA lesions may be generated per day in a single cell, and this has been linked to a wide variety of diseases, including cancer. However, it is puzzling why potentially mutagenic DNA modifications, occurring at a similar level in different organs/tissue, may lead to organ/tissue specific cancers, or indeed non-malignant disease – what is the basis for this differential response? We suggest that it is perhaps the precise location of damage, within the genome, that is a key factor. Finally, we draw attention to the requirement for reliable methods for identification and quantification of DNA adducts/modifications, and stress the need for these assays to be fully validated. Once these prerequisites are satisfied, measurement of DNA modifications may be helpful as a clinical parameter for treatment monitoring, risk group identification and development of prevention strategies.     

Do Antarctic populations represent local or widespread phylogenetic and ecological lineages? Complicated fate of bipolar moss concepts with <Emphasis Type="Italic">Drepanocladus longifolius</Emphasis> as a case study

Bipolar moss species, extreme examples of large and disjunctive geographical ranges, belong to the most important components of the vegetation cover in Antarctica. Their appropriate taxonomic interpretations are essential for fine-scale biogeographical considerations but our knowledge on their phylogenetic affinities still remains limited. Here, we address the history of the Antarctic populations of Drepanocladus longifolius, a moss species widely considered a bipolar taxon. First, based on a worldwide sampling, we verify its assumed bipolar status. Then, we ask whether its current Antarctic populations have persisted throughout at least the last major glaciation in situ or, alternatively, are they a recent, postglacial element. Phylogenetic analyses together with investigation of morphological characters were used to infer relationships among accessions representing the whole distribution range of the taxon. Morphological and phylogenetic data strictly segregated specimens from the Southern and the Northern Hemisphere, with an exception of Australian populations that were included in the latter group. The Antarctic, sub-Antarctic and South American populations belong to a single clade and are treated as D. longifolius s. str., which consequently is not a bipolar taxon. All Northern Hemisphere populations and the ones from Australia represent D. capillifolius, a lately neglected taxon, which thus displays a true bipolar distribution pattern but not comprising the Antarctic and South American populations. These results provide a sound example of key importance of a detailed taxonomical and phylogenetic treatment as a basis for biogeographical studies. In D. longifolius s. str., a homogeneity in all non-coding nuclear and plastid regions was observed throughout the whole range. It contrasted with the presence of one missense mutation in the rps4 gene in all examined Antarctic populations, which correlated with their distinct ecological situation. While identity of non-coding sequences in D. longifolius populations indicates a postglacial recruitment of extant Antarctic populations from extra-regional locations rather than a long-term in situ survival, the mutation in the rps4 gene may be hypothesised to reflect an adaptive response of plants to the Antarctic environment.