DNA methylation of Quercus robur L. plumules following cryo-pretreatment and cryopreservation

Pedunculate oak (Quercus robur) is an ecologically and economically important forest tree species which produces seeds that are classified as recalcitrant. Thus, cryopreservation of seed meristems is a method for long-term preservation of this germplasm in gene banks. During cryopreservation, many factors, such as desiccation, cryoprotection and cooling/rewarming, can induce stress in the frozen meristems. In this study, in vitro survival and the global DNA methylation level of plumules after cryoprotection, desiccation and cryostorage was evaluated. Results indicated that both desiccation and storage in liquid nitrogen have negligible influence on DNA methylation status of Q. robur plumules. These findings support the cryopreservation of plumules as an appropriate method for conservation of Q. robur germplasm.     

Embryonic and adult isoforms of XLAP2 form microdomains associated with chromatin and the nuclear envelope

Laminin-associated polypeptide 2 (LAP2) proteins are alternatively spliced products of a single gene; they belong to the LEM domain family and, in mammals, locate to the nuclear envelope (NE) and nuclear lamina. Isoforms lacking the transmembrane domain also locate to the nucleoplasm. We used new specific antibodies against the N-terminal domain of Xenopus LAP2 to perform immunoprecipitation, identification and localization studies during Xenopus development. By immunoprecipitation and mass spectrometry (LC/MS/MS), we identified the embryonic isoform XLAP2??, which was downregulated during development similarly to XLAP2??. Embryonic isoforms XLAP2?? and XLAP2?? were located in close association with chromatin up to the blastula stage. Later in development, both embryonic isoforms and the adult isoform XLAP2?? were localized in a similar way at the NE. All isoforms colocalized with lamin B2/B3 during development, whereas XLAP2?? was colocalized with lamin B2 and apparently with the F/G repeat nucleoporins throughout the cell cycle in adult tissues and culture cells. XLAP2?? was localized in clusters on chromatin, both at the NE and inside the nucleus. Embryonic isoforms were also localized in clusters at the NE of oocytes. Our results suggest that XLAP2 isoforms participate in the maintenance and anchoring of chromatin domains to the NE and in the formation of lamin B microdomains.     

Magnesium,Calcium, and Trace Elements Excretion in 24-h Urine

Urine is a clinical specimen often used in medical diagnostics for monitoring of elements concentrations and kidneys function. We determined the contents of magnesium (Mg), calcium (Ca), zinc (Zn), copper (Cu), iron (Fe), lead (Pb), and cadmium (Cd) in 74 samples of 24-h urine (from 46 women and 28 men). The measurements were realized by the atomic absorption spectrometry (AAS) with atomization in the flame (FAAS) and in the graphite furnace (GFAAS). The received results were the subject of statistical analysis including the sex and age of volunteers. Moreover, correlations between the elements and the relationships between age and amounts of excreted elements with urine were tested. We found the statistically significant higher content of Zn in men’s urine than in women^’s one. Moreover, both adult women and men (>18 years) excreted much more Ca in urine in comparison to young subjects. Only in case of Pb the significant positive correlation between its amount in 24-h urine of all donors and age was stated. The correlation analysis has shown the significant positive relationships between Ca–Mg, Ca–Fe, Mg–Fe, Cu–Fe, Cu–Cd, Fe–Cd, and Pb–Cd in total samples of urine. Basing on our results, we concluded that the gender and age of donors may impact on the elemental status of 24-h urine.     

Lymphatic vessel invasion detected by the endothelial lymphatic marker D2-40 (podoplanin) is predictive of regional lymph node status and an independent prognostic factor in patients with resected esophageal cancer

The discovery of markers to lymphatic endothelial cells and the development of novel antibodies to these markers have brought increasing attention to the lymphatics and progress in the understanding of lymphangiogenesis and cancer metastasis. In this study, we investigate the presence of lymphatic vessel invasion (LVI) detected by D2-40 immunohistochemical staining in resected esophageal cancer and correlated with clinicopathologic data and patient survival. Sixty nine patients, who had a primary resection of esophageal cancer, were analyzed by univariate and multivariate logistic regression, and univariate and multivariate survival analysis. The total rate of LVI was 72% (50/69). Positive LVI was significantly correlated with lymph node metastasis (p < 0.001), tumor size (p < 0.001), histological grading (p = 0.017), tumor depth (p = 0.001), and stage (p < 0.001). Multivariate logistic analysis identified LVI (p = 0.036) as a predictor of regional lymph node metastasis. On univariate survival analysis, patients with LVI had a significantly shorter disease-free survival, cancer-specific survival and overall survival. Multivariate analysis proved that LVI diagnosed by D2-40 is an independent prognostic factor of both disease-free survival (p = 0.04) and overall survival (p = 0.032) in resected esophageal cancer. These results show that LVI assessment identifies patients at high risk for regional lymph node metastasis and that LVI is an independent prognostic factor in patients with esophageal cancer.     

Systemic changes in photosynthesis and reactive oxygen species homeostasis in shoots of Arabidopsis thaliana infected with the beet cyst nematode Heterodera schachtii

Photosynthetic efficiency and redox homeostasis are important for plant physiological processes during regular development as well as defence responses. The second‐stage juveniles of Heterodera schachtii induce syncytial feeding sites in host roots. To ascertain whether the development of syncytia alters photosynthesis and the metabolism of reactive oxygen species (ROS), chlorophyll a fluorescence measurements and antioxidant responses were studied in Arabidopsis thaliana shoots on the day of inoculation and at 3, 7 and 15 days post‐inoculation (dpi). Nematode parasitism caused an accumulation of superoxide and hydrogen peroxide molecules in the shoots of infected plants at 3 dpi, probably as a result of the observed down‐regulation of antioxidant enzymes. These changes were accompanied by an increase in RNA and lipid oxidation markers. The activities of antioxidant enzymes were found to be enhanced on infection at 7 and 15 dpi, and the content of anthocyanins was elevated from 3 dpi. The fluorescence parameter R_fd, defining plant vitality and the photosynthetic capacity of leaves, decreased by 11% only at 7 dpi, and non‐photochemical quenching (NPQ), indicating the effectiveness of photoprotection mechanisms, was about 16% lower at 3 and 7 dpi. As a result of infection, the ultrastructure of chloroplasts was changed (large starch grains and plastoglobules), and more numerous and larger peroxisomes were observed in the mesophyll cells of leaves. We postulate that the joint action of antioxidant enzymes/molecules and photochemical mechanisms leading to the maintenance of photosynthetic efficiency promotes the fine‐tuning of the infected plants to oxidative stress induced by parasitic cyst nematodes.     

Impact of environmental diversity of hunting complexes in the Lublin region on ontogenetic quality indicators in roe deer (<Emphasis Type="Italic">Capreolus capreolus</Emphasis>)

Populations of game are not confined to single ecosystems but function within higher-order units, e.g. ecological landscape. The basis for the establishment of the hunting complexes was the assumption that the existing game hunting grounds, i.e. the basic units implementing game management, are too small and do not cover the natural areas inhabited by game populations. Roe deer are flexible species and easily adapt to various site conditions, so they inhabit many different habitats, from large forest complexes, through small in-field tree stands and shrubs, to treeless grounds and field monocultures. The aim of the study was to determine a possible impact of environmental conditions prevailing in the hunting complexes of the Regional Directorate of State Forests (RDLP in Lublin) on the ontogenetic quality of roe deer. The study was conducted on 518 European roe deer (Capreolus capreolus) aged from 4 to 7 years (379 bucks and 139 does) harvested within hunting seasons 2010/2011–2013/2014. The results have shown that animals originating from areas with greater forest cover and denser stands are characterised by lower values of the mean ontogenetic quality parameters (carcase weight, kidney fat index, chest girth, weight of antlers) in comparison with animals from typical agricultural areas with fragmented forest complexes. These results indicate that, even in the case of such a eurytopic species as the roe deer, the ontogenetic quality differs between individual hunting complexes. The study has proved that strategies for hunting management of the roe deer should take into account the impact of the landscape structure, which provides a rationale behind creation of hunting complexes.     

Variation in fish growth characteristics along a river course

Variation in the growth patterns of roach, Rutilus rutilus (L.), pike, Esox lucius L., and chub, Leuciscus cephalus (L.) was examined along the upper Warta River, where human impact (mostly pollution) has influenced the longitudinal zonation on the fish assemblage. Significant differences were found in the exponent of weight-length relationships for roach and chub populations occupying different zones of the river, but no such variation was observed in pike. Moreover, pike growth was isometric, whereas roach and chub grew allometrically, with regression coefficients (slope) above 3. Although the length-at-age data were similar for each zone, the von Bertalanffy parameters (L _inf, K and t ₀) suggest that there may exist some inter-zone variation in the overall growth patterns of these species. All the species grew better in the zone where the index of relative abundance (relating dominance of a particular species to its maximum abundance in river system) achieved its highest value. The results suggest that a relative abundance index expressed in this way can be a good index of habitat quality.     

Immunohistochemical characteristics of nerve fibres supplying the porcine vas deferens. A colocalisation study

 Double-labelling immunofluorescence was used to investigate the coexistence of the catecholamine-synthesising enzymes, tyrosine hydroxylase and dopamine-β-hydroxylase and several neuropeptides including neuropeptide Y, vasoactive intestinal polypeptide, Leu⁵-enkephalin, somatostatin, calcitonin gene-related peptide and substance P in nerve fibres supplying the vas deferens in juvenile and adult pigs. The study has revealed three major populations of nerve terminals innervating the organ: (1) noradrenergic fibres; (2) non-noradrenergic (putative cholinergic) fibres containing vasoactive intestinal polypeptide, neuropeptide Y and somatostatin, supplying almost exclusively the lamina propria; and (3) non-noradrenergic, presumably sensory fibres, containing calcitonin gene-related peptide and substance P. The population of noradrenergic nerves can be divided into three subpopulations: a somatostatin-containing, a Leu⁵-enkephalin-containing and a subpopulation immunonegative to the peptides investigated, in descending order of magnitude. Coexistence patterns of the substances existing within nerve fibres supplying the vas deferens blood vessels are clearly different from those found in nerve fibres innervating the organ wall. The majority of the noradrenergic fibres associated with blood vessels contain neuropeptide Y only, while non-noradrenergic perivascular nerves contain predominantly vasoactive intestinal polypeptide. The possibility of different sources of origin of the particular nerve fibre subpopulations supplying the porcine vas deferens and its blood vessels is discussed. Accepted: 23 October 1996     

Developmental response of carp,Cyprinus carpio,larvae fed different foods or starved

Synopsis The developmental interval, body length, dry and wet weight were determined in carp larvae during the first 2 weeks of exogenous feeding. Six feeding groups were reared at 26°C; four groups received different artificial diets, whereas zooplankton-fed larvae and unfed larvae served as controls. It was found that larval developmental rate can be modified by the type of food. The diet affects the relationship between developmental step and weight. The effect of diet on the relationship between development and length becomes manifest at older steps (after longer feeding the same diet). A threshold point, after which satisfactory results of feeding artificial diets to carp larvae can be expected, depends not only on body size but also on the degree of development.     

On the origin of surface imposed anisotropic growth of salicylic and acetylsalicylic acids crystals during droplet evaporation

In this paper droplet evaporative crystallization of salicylic acid (SA) and acetylsalicylic acid (ASA) crystals on different surfaces, such as glass, polyvinyl alcohol (PVA), and paraffin was studied. The obtained crystals were analyzed using powder X-ray diffraction (PXRD) technique. In order to better understand the effect of the surface on evaporative crystallization, crystals deposited on glass were scraped off. Moreover, evaporative crystallization of a large volume of solution was performed. As we found, paraffin which is non-polar surface promotes formation of crystals morphologically similar to those obtained via bulk evaporative crystallization. On the other hand, when crystallization is carried out on the polar surfaces (glass and PVA), there is a significant orientation effect. This phenomenon is manifested by the reduction of the number of peaks in PXRD spectrum recorded for deposited on the surface crystals. Noteworthy, reduction of PXRD signals is not observed for powder samples obtained after scraping crystals off the glass. In order to explain the mechanism of carboxylic crystals growth on the polar surfaces, quantum-chemical computations were performed. It has been found that crystal faces of the strongest orientation effect can be characterized by the highest surface densities of intermolecular interactions energy (IIE). In case of SA and ASA crystals formed on the polar surfaces the most dominant faces are characterized by the highest adhesive and cohesive properties. This suggests that the selection rules of the orientation effect comes directly from surface IIE densities.