Application of DNA marker systems to test for genetic imprints of habitat fragmentation in Juniperus communis L. on different spatial and temporal scales-Integration of scientific knowledge into conservation measures

The formerly continuous and widespread juniper (Juniperus communis L.) populations are currently divided into small and fragmented relics in numerous European countries. Additionally, many of these populations suffer from an absence of natural regeneration and consist predominantly of senescent individuals. In order to maintain juniper as a valuable element of the cultural landscape in Europe, I considered a restoration management to be indispensable. The goal of the present thesis is based on this consideration. Using different DNA marker systems, I firstly tested various juniper populations on different spatial and temporal scales for potential imprints of habitat fragmentation. Afterwards my intention was to evaluate the analysed populations on the regional scale in terms of nature conservation and to develop a scientifically based conservation management plan, which should focus on planting activities. In a Europe-wide study I used an AFLP marker approach to reconstruct aspects of the biogeographic history of juniper and to detect potential distinct genetic lineages. Those lineages are supposed to delineate geographic regions within which plant material can be interchanged. The genetic analysis revealed no distinct genetic lineages. Along with other scientific findings about juniper the results point to a glacial persistence of juniper in Central Europe. I suppose that during the last glacial period, this species managed to survive in several small and suitable habitats, which were probably diffusely scattered and permanently changing. Moreover, I hypothesise that recurrent fragmentation and founder events since the last glacial maximum (LGM) up until today are highly likely to have occurred in this species. On a regional scale, i.e. in the Rhenish Uplands (RU) (West Germany), I used nSSR markers to gain insights into the genetic structure and variation of eight relict juniper populations. Such knowledge is necessary for planting activities in order to prevent negative effects in the respective populations. At the same time, I tested these populations for genetic imprints of the recent habitat fragmentation. The investigated nSSR loci in juniper were characterised within the scope of this thesis. A detailed validation of the newly developed nSSR markers is presented. In addition, I performed a case study by investigating the genetic diversity and differentiation of different pollen clouds, which have become reproductively effective in the filial generations (embryos). For this purpose a specific computer software was developed. Next, a palynological study was conducted to determine physical pollen flow distances of juniper pollen grains. In terms of plantings I assume that such data is relevant for the spatial organisation of already existing juniper individuals and the respective plant material. Considerably high levels of genetic diversity and an absence of recent genetic bottlenecks in all populations as well as an absence of an isolation- by-distance effect led me to the assumption that the current habitat fragmentation has not yet affected the genetic diversity in the investigated juniper populations. Instead, I postulate that the genetic diversity and differentiation have been ‘frozen’ since the recent fragmentation started. The genetic diversity of the filial generation is not reduced in comparison to the adult generation, although the palynological study points to locally restricted pollen flow distances. After defining a ‘Leitbild’ for viable juniper populations based on widely accepted population ecological and genetic theories, I used the genetic results to evaluate the analysed populations of the RU in terms of nature conservation. The reasons why this evaluation was not satisfactory are discussed in detail. Further on, I commented critically on the evaluation criteria of the ‘Leitbild’ and their respective quality demands with regard to the life-history traits of juniper and its biogeographic history as presented within this thesis. Based on the presented genetic results and on the apparent absence of natural regeneration in all populations it remains uncertain whether the current habitat fragmentation will affect the genetic diversity and structure of the eight populations deleteriously in the future. However, if juniper will not start with natural regeneration again, this will certainly lead to an extinction of the respective populations because without substitution, senescent individuals will gradually die off. Thus, in the distant future juniper will probably become extinct in areas where it does not regenerate naturally. Therefore, I developed a sustainable, demographically and genetically substantiated restoration management plan as a final outcome of this thesis. It is based on the genetic analysis presented here and on expert knowledge, and it includes guidelines and recommendations concerning the collection of plant material, its treatment in the greenhouse and plantings in the field

Molecular mechanism of topoisomerase poisoning by the peptide antibiotic albicidin

The peptide antibiotic albicidin is a DNA topoisomerase inhibitor with low-nanomolar bactericidal activity towards fluoroquinolone-resistant Gram-negative pathogens. However, its mode of action is poorly understood. We determined a 2.6 Å resolution cryoelectron microscopy structure of a ternary complex between Escherichia coli topoisomerase DNA gyrase, a 217 bp double-stranded DNA fragment and albicidin. Albicidin employs a dual binding mechanism where one end of the molecule obstructs the crucial gyrase dimer interface, while the other intercalates between the fragments of cleaved DNA substrate. Thus, albicidin efficiently locks DNA gyrase, preventing it from religating DNA and completing its catalytic cycle. Two additional structures of this trapped state were determined using synthetic albicidin analogues that demonstrate improved solubility, and activity against a range of gyrase variants and E. coli topoisomerase IV. The extraordinary promiscuity of the DNA-intercalating region of albicidins and their excellent performance against fluoroquinolone-resistant bacteria holds great promise for the development of last-resort antibiotics

Redescription of Milnesium alpigenum Ehrenberg, 1853 (Tardigrada: Apochela) and a description of Milnesium inceptum sp. nov., a tardigrade laboratory model

Intra- and interspecific variability, being at the very core of alpha taxonomy, has been a long-standing topic of debate among tardigrade taxonomists. Early studies tended to assume that tardigrades exhibit wide intraspecific variation. However, with more careful morphological studies, especially those incorporating molecular tools that allow for an independent verification of species identifications based on phenotypic traits, we now recognise that ranges of tardigrade intraspecific variability are narrower, and that differences between species may be more subtle than previously assumed. The taxonomic history of the genus Milnesium, and more specifically that of the nominal species, M. tardigradum described by Doyère in 1840, is a good illustration of the evolution of views on intraspecific variability in tardigrades. The assumption of wide intraspecific variability in claw morphology led Marcus (1928) to synonymise two species with different claw configurations, M. alpigenum and M. quadrifidum, with M. tardigradum. Currently claw configuration is recognised as one of the key diagnostic traits in the genus Milnesium, and the two species suppressed by Marcus have recently been suggested to be valid. In this study, we clarify the taxonomic status of M. alpigenum, a species that for nearly a century was considered invalid. We redescribe M. alpigenum, using a population collected from the locus typicus, by the means of integrative taxonomy, i.e. including light microscopy, scanning electron microscopy, ontogenetic observations, and genetic barcoding. Moreover, the redescription of M. alpigenum allowed us to verify the uncertain taxonomic status of two popular laboratory models that were originally considered to be M. tardigradum; though one was recently reidentified as M. cf. alpigenum. Our analysis showed that both laboratory strains, despite being morphologically and morphometrically nearly identical to M. alpigenum, in fact represent a new species, M. inceptum sp. nov. The two species, being disnguishable only by statistical morphometry and/or DNA sequences, are the first example of pseudocryptic species in tardigrades

Przydatność biotestów ekotoksykologicznych do oceny stanu biologicznego wód na przykładzie zbiornika zaporowego w Goczałkowicach

Implementation of ecotoxicological biotests for monitoring of water quality in water reservoirs is one of the aims of the project “Integrated system supporting management and protection of water reservoir (ZiZOZap)”. The project is executed within The Programme Innovative Economy under National Strategic Reference Framework as a part of introducing Water Framework Directive. The model object of the Project is Goczalkowice Water Reservoir. The main purposes of the Project are maintaining a good condition of the Reservoir for future and prevent processes of its aging and degradation. The final product of the Project will be an informational system based on the results of environmental monitoring, numerical models of reservoir and relative management scenarios. The system will allow permanent assessment of current reservoir condition as well as prediction of its short time and long time alterations. In consequence, these may lead to rationale decisions referring to reservoir management. Since July 2010 a battery of standardized commercial biotests is applied for water quality monitoring in Goczalkowice Reservoir. The obtained results are referred to the standards set by governmental and EC ordinances. We assume the biotests would be useful in constructing detailed scenarios of the reservoir management.

Współczesne wyzwania teorii i praktyki bezpieczeństwa

Z wprowadzenia: "Nauki o bezpieczeństwie mają niespełna sześcioletnią historię, kiedy to 8 sierpnia 2011 roku minister Nauki i Szkolnictwa Wyższego ustanowiła je swoim rozporządzeniem jako nawą dyscyplinę naukową w dziedzinie nauk społecznych nowo utworzonego obszaru wiedzy np. nauki społeczne. Samo zaś bezpieczeństwo należy do jednej z najbardziej elementarnych potrzeb człowieka. Jest podstawowym prawem i jego potrzebą, sprowadza się ono do eliminowania sytuacji zagrażających człowiekowi i jego dobrom. Poczucie bezpieczeństwa jest jedną z dwóch najistotniejszych potrzeb człowieka warunkującą dalszy jego rozwój i funkcjonowanie w społeczeństwie. Bezpieczeństwo ma umożliwić prawidłowy rozwój społeczny przy przestrzeganiu zasad współżycia społecznego."(...)Publikacja powstała w ramach projektu badawczego Krakowskiej Akademii im. Andrzeja Frycza Modrzewskiego nr WNoB/DS/3/2015-KO

Pregnancy, delivery and puerperium in a patient with lysinuric protein intolerance – a case report

The paper presents the course of pregnancy, delivery and early postpartum period in a 23-year-old woman with lysinuric protein intolerance (LPI). The pregnancy was uneventful and resulted in a caesarean birth to a healthy baby at 37 weeks gestation. Nevertheless, the course of pregnancy in women with LPI is associated with a significantly increased risk of serious complications, including acute hyperammonemia, preeclampsia and postpartum bleeding, as well as fetus intrauterine growth retardation. In many cases, intensive metabolic monitoring and a proper diet with protein limitation and appropriate amino acids supplementation may significantly reduce the risk for both the mother and the newborn

Effects of ATP7A overexpression in mice on copper transport and metabolism in lactation and gestation

Placentae and mammary epithelial cells are unusual in robustly expressing two copper "pumps", ATP7A and B, raising the question of their individual roles in these tissues in pregnancy and lactation. Confocal microscopic evidence locates ATP7A to the fetal side of syncytiotrophoblasts, suggesting a role in pumping Cu towards the fetus; and to the basolateral (blood) side of lactating mammary epithelial cells, suggesting a role in recycling Cu to the blood. We tested these concepts in wild-type C57BL6 mice and their transgenic counterparts that expressed hATP7A at levels 10-20× those of endogenous mAtp7a. In lactation, overexpression of ATP7A reduced the Cu concentrations of the mammary gland and milk ~50%. Rates of transfer of tracer (64)Cu to the suckling pups were similarly reduced over 30-48 h, as was the total Cu in 10-day -old pups. During the early and middle periods of gestation, the transgenic litters had higher Cu concentrations than the wild-type, placental Cu showing the reverse trend; but this difference was lost by the first postnatal day. The transgenic mice expressed ATP7A in some hepatocytes, so we investigated the possibility that metalation of ceruloplasmin (Cp) might be enhanced. Rates of (64)Cu incorporation into Cp, oxidase activity, and ratios of holo to apoceruloplasmin were unchanged. We conclude that in the lactating mammary gland, the role of ATP7A is to return Cu to the blood, while in the placenta it mediates Cu delivery to the fetus and is the rate-limiting step for fetal Cu nutrition during most of gestation in mice

Clinical complexity and impact of the ABC (Atrial fibrillation Better Care) pathway in patients with atrial fibrillation: a report from the ESC-EHRA EURObservational Research Programme in AF General Long-Term Registry

Background: Clinical complexity is increasingly prevalent among patients with atrial fibrillation (AF). The ‘Atrial fibrillation Better Care’ (ABC) pathway approach has been proposed to streamline a more holistic and integrated approach to AF care; however, there are limited data on its usefulness among clinically complex patients. We aim to determine the impact of ABC pathway in a contemporary cohort of clinically complex AF patients. Methods: From the ESC-EHRA EORP-AF General Long-Term Registry, we analysed clinically complex AF patients, defined as the presence of frailty, multimorbidity and/or polypharmacy. A K-medoids cluster analysis was performed to identify different groups of clinical complexity. The impact of an ABC-adherent approach on major outcomes was analysed through Cox-regression analyses and delay of event (DoE) analyses. Results: Among 9966 AF patients included, 8289 (83.1%) were clinically complex. Adherence to the ABC pathway in the clinically complex group reduced the risk of all-cause death (adjusted HR [aHR]: 0.72, 95%CI 0.58–0.91), major adverse cardiovascular events (MACEs; aHR: 0.68, 95%CI 0.52–0.87) and composite outcome (aHR: 0.70, 95%CI: 0.58–0.85). Adherence to the ABC pathway was associated with a significant reduction in the risk of death (aHR: 0.74, 95%CI 0.56–0.98) and composite outcome (aHR: 0.76, 95%CI 0.60–0.96) also in the high-complexity cluster; similar trends were observed for MACEs. In DoE analyses, an ABC-adherent approach resulted in significant gains in event-free survival for all the outcomes investigated in clinically complex patients. Based on absolute risk reduction at 1 year of follow-up, the number needed to treat for ABC pathway adherence was 24 for all-cause death, 31 for MACEs and 20 for the composite outcome. Conclusions: An ABC-adherent approach reduces the risk of major outcomes in clinically complex AF patients. Ensuring adherence to the ABC pathway is essential to improve clinical outcomes among clinically complex AF patients