Daunorubicin induces cell death via activation of apoptotic signalling pathway and inactivation of survival pathway in muscle-derived stem cells

Daunorubicin (as well as other anthracyclines) is known to be toxic to heart cells and other cells in organism thus limiting its applicability in human cancer therapy. To investigate possible mechanisms of daunorubicin cytotoxicity, we used stem cell lines derived from adult rabbit skeletal muscle. Recently, we have shown that daunorubicin induces apoptotic cell death in our cell model system and distinctly influences the activity of MAP kinases. Here, we demonstrate that two widely accepted antagonistic signalling pathways namely proapoptotic JNK and prosurvival PI3K/AKT participate in apoptosis. Using the Western blot method, we observed the activation of JNK and phosphorylation of its direct target c-Jun along with inactivation of AKT and its direct target GSK in the course of programmed cell death. By means of small-molecule kinase inhibitors and transfection of cells with the genes of the components of these pathways, c-Jun and AKT, we confirm that JNK signalling pathway is proapoptotic, whereas AKT is antiapoptotic in daunorubicin-induced muscle cells. These findings could contribute to new approaches which will result in less toxicity and fewer side effects that are currently associated with the use of daunorubicin in cancer therapies.     

Different Antibiotic Resistance and Sporulation Properties within Multiclonal Clostridium difficile PCR Ribotypes 078, 126, and 033 in a Single Calf Farm

Clostridium difficile strains were sampled periodically from 50 animals at a single veal calf farm over a period of 6 months. At arrival, 10% of animals were C. difficile positive, and the peak incidence was determined to occur at the age of 18 days (16%). The prevalence then decreased, and at slaughter, C. difficile could not be isolated. Six different PCR ribotypes were detected, and strains within a single PCR ribotype could be differentiated further by pulsed-field gel electrophoresis (PFGE). The PCR ribotype diversity was high up to the animal age of 18 days, but at later sampling points, PCR ribotype 078 and the highly related PCR ribotype 126 predominated. Resistance to tetracycline, doxycycline, and erythromycin was detected, while all strains were susceptible to amoxicillin and metronidazole. Multiple variations of the resistance gene tet(M) were present at the same sampling point, and these changed over time. We have shown that PCR ribotypes often associated with cattle (ribotypes 078, 126, and 033) were not clonal but differed in PFGE type, sporulation properties, antibiotic sensitivities, and tetracycline resistance determinants, suggesting that multiple strains of the same PCR ribotype infected the calves and that calves were likely to be infected prior to arrival at the farm. Importantly, strains isolated at later time points were more likely to be resistant to tetracycline and erythromycin and showed higher early sporulation efficiencies in vitro, suggesting that these two properties converge to promote the persistence of C. difficile in the environment or in hosts.     

Integrating ongoing biodiversity monitoring: potential benefits and methods

Halting the loss of biodiversity comes along with the need to quantify biodiversity composition and dynamics at large spatial and temporal scales. Highly standardized, international monitoring networks would be ideal, but they do not exist yet. If we are to assess changes in biodiversity now, combining output available from ongoing monitoring initiatives is the only option. However, integration of biodiversity information across schemes is still very poorly developed. In this paper, we outline practical issues to be considered when planning to combine existing monitoring information. First, we provide an overview of avenues for integration along the four dimensions that characterize a monitoring design: sample size, biological coverage, spatial coverage and temporal coverage. We also emphasize that complementarity in monitoring targets across schemes enables to describe complex processes of biodiversity dynamics, e.g. through relating species traits to the impacts of environmental changes. Second, we review some methods to overcome differences in designs among monitoring schemes, such as site selection, post-stratification and measurement error. Finally, we point out some commonly used statistical methods that are at hand for combining data or parameter estimates. We especially emphasize the possible levels of data integration (raw data, parameter estimates, or effect size estimates), and the largely under-exploited potential of meta-analysis methods and weighted analyses. This contribution aims to bolster the practice and use of integration of ongoing monitoring initiatives for biodiversity assessment.     

Habitat monitoring in Europe: a description of current practices

Monitoring of biodiversity at the level of habitats is becoming increasingly common. Here we describe current practices in habitat monitoring based on 150 schemes in Europe. Most schemes were initiated after 1990 in response to EU nature directives or habitat management/restoration actions, with funding mostly from European or national sources. Schemes usually monitor both the spatial distribution and the quality of the habitats, and they frequently collect data on environmental parameters and potential causes of changes. Many schemes are local or regional rather than national or international in scope, and sampling effort varies greatly across spatial and temporal scales. Experimental design is used in half of the schemes, however, data are rarely analysed by advanced statistics. Most schemes require two months or less per year in manpower and are typically run by professionals rather than by volunteers. Estimated salaries plus equipment costs average 650,000 Euro per year per scheme, and add up to 80 million Euros annually. Costs are particularly high for schemes based on European or international law and for schemes funded by European or national sources. Costs are also high in schemes in which sampling sites are selected subjectively rather than based on sampling theory, and in schemes that do not use field mapping or remote sensing to document spatial variation in habitats. Our survey demonstrates promising developments in European habitat monitoring but also underlines the need for better spatial coverage, documentation of spatial variaton, improved sampling design and advanced data analysis. Such improvements are essential if we are to judge progress towards the 2010 biodiversity targets.     

Measuring the Quality of Life of Visually Impaired Children: First Stage Psychometric Evaluation of the Novel VQoL_CYP Instrument

Purpose

To report piloting and initial validation of the VQoL_CYP, a novel age-appropriate vision-related quality of life (VQoL) instrument for self-reporting by children with visual impairment (VI).

Methods

Participants were a random patient sample of children with VI aged 10–15 years. 69 patients, drawn from patient databases at Great Ormond Street Hospital and Moorfields Eye Hospital, United Kingdom, participated in piloting of the draft 47-item VQoL instrument, which enabled preliminary item reduction. Subsequent administration of the instrument, alongside functional vision (FV) and generic health-related quality of life (HRQoL) self-report measures, to 101 children with VI comprising a nationally representative sample enabled further item reduction and evaluation of psychometric properties using Rasch analysis. Construct validity was assessed through Pearson correlation coefficients.

Results

Item reduction through piloting (8 items removed for skewness and individual item response pattern) and validation (1 item removed for skewness and 3 for misfit in Rasch) produced a 35-item scale, with fit values within acceptable limits, no notable differential item functioning, good measurement precision, ordered response categories and acceptable targeting in Rasch. The VQoL_CYP showed good construct validity, correlating strongly with HRQoL scores, moderately with FV scores but not with acuity.

Conclusions

Robust child-appropriate self-report VQoL measures for children with VI are necessary for understanding the broader impacts of living with a visual disability, distinguishing these from limited functioning per se. Future planned use in larger patient samples will allow further psychometric development of the VQoL_CYP as an adjunct to objective outcomes assessment.     

A review and a framework for the integration of biodiversity monitoring at the habitat level

The monitoring of biodiversity at the level of habitats is becoming widespread in Europe and elsewhere as countries establish national habitat monitoring systems and various organisations initiate regional and local schemes. Parallel to this growth, it is increasingly important to address biodiversity changes on large spatial (e.g. continental) and temporal (e.g. decade-long) scales, which requires the integration of currently ongoing monitoring efforts. Here we review habitat monitoring and develop a framework for integrating data or activities across habitat monitoring schemes. We first identify three basic properties of monitoring activities: spatial aspect (explicitly spatial vs. non-spatial), documentation of spatial variation (field mapping vs. remote sensing) and coverage of habitats (all habitats or specific habitats in an area), and six classes of monitoring schemes based on these properties. Then we explore tasks essential for integrating schemes both within and across the major classes. Finally, we evaluate the need and potential for integration of currently existing schemes by drawing on data collected on European habitat monitoring in the EuMon project. Our results suggest a dire need for integration if we are to measure biodiversity changes across large spatial and temporal scales regarding the 2010 target and beyond. We also make recommendations for an integrated pan-European habitat monitoring scheme. Such a scheme should be based on remote sensing to record changes in land cover and habitat types over large scales, with complementary field mapping using unified methodology to provide ground truthing and to monitor small-scale changes, at least in habitat types of conservation importance.     

Phylogeny of the cave shrimp Troglocaris: Evidence of a young connection between Balkans and Caucasus

The remarkably discontinuous distribution of the cave shrimp genus Troglocaris in South France, West Balkans, and West Caucasus has long been considered a biogeographic enigma. To solve it, its phylogeny was reconstructed by analyzing sequences from two mitochondrial (cytochrome oxidase I and 16S rRNA) and one nuclear gene (28S rRNA) using maximum likelihood, parsimony and Bayesian inference. The genus was found to be polyphyletic because the French taxon T. inermis had no direct common ancestry with other Troglocaris taxa but was sister to the epigean freshwater atyid Dugastella valentina. All other Troglocaris species constituted a well-supported monophylum, the second cave shrimp genus Spelaeocaris nested within. The monophylum had a well-defined structure: (1) a clade restricted to the Dinaric area of the Western Balkans containing the type species T. anophthalmus along with some unnamed species, and (2) a geographically mixed clade split between the Caucasian T. kutaissiana species complex on one, and T. hercegovinensis, S. pretneri, plus an unnamed taxon on the other side. It was surprising to find the dichotomy between the Caucasian and one of the West-Balkan lineages so low in the phylogenetic hierarchy of the genus. Taking into account molecular rates of other decapods, we tentatively dated this split at 6-11 Myr. This time is in agreement with the brackish and freshwater phase of the Paratethys thus allowing for a freshwater common ancestor of Caucasian and Dinaric cave shrimps. This would weaken the marine relicts hypothesis that has often been invoked to explain the distribution of freshwater cave species with close marine relatives.     

Model‐based optimization and pO2 control of fed‐batch Escherichia coli and Saccharomyces cerevisiae cultivation processes

A model‐based approach for optimization and cascade control of dissolved oxygen partial pressure (pO₂) and maximization of biomass in fed‐batch cultivations is presented. The procedure is based on the off‐line model‐based optimization of the optimal feeding rate profiles and the subsequent automatic pO₂ control using a proposed cascade control technique. During the model‐based optimization of the process, feeding rate profiles are optimized with respect to the imposed technological constraints (initial and maximal cultivation volume, cultivation time, feeding rate range, maximal oxygen transfer rate and pO₂ level). The cascade pO₂ control is implemented using activation of cascades for agitation, oxygen enrichment, and correction of the preoptimized feeding rate profiles. The proposed approach is investigated in two typical fed‐batch processes with Escherichia coli and Saccharomyces cerevisiae. The obtained results show that it was possible to achieve sufficiently high biomass levels with respect to the given technological constraints and to improve controllability of the investigated processes.