Sphingosine kinase 1 and 2 regulate the capacity of mesangial cells to resist apoptotic stimuli in an opposing manner

Abstract Sphingosine kinases (SKs) are key enzymes regulating the production of sphingosine-1-phosphate (S1P), which determines important cell responses including cell growth and death. Here we show that renal mesangial cells isolated from wild-type, SK-1(-/-), and SK-2(-/-) mice show a differential response to apoptotic stimuli. Wild-type mesangial cells responded to staurosporine with increased DNA fragmentation and caspase-3 processing, which was enhanced in SK-1(-/-) cells. In contrast, SK-2(-/-) cells were highly resistant to staurosporine-induced apoptosis. Furthermore, the basal phosphorylation and activity of the anti-apoptotic protein kinase B (PKB) and of its substrate Bad were decreased in SK-1(-/-) but not in SK-2(-/-) cells. Upon staurosporine treatment, phosphorylation of PKB and Bad decreased in wild-type and SK-1(-/-) cells, but remained high in SK-2(-/-) cells. In addition, the anti-apoptotic Bcl-X(L) was significantly upregulated in SK-2(-/-) cells, which may further contribute to the protective state of these cells. In summary, our data show that SK-1 and SK-2 have opposite effects on the capacity of mesangial cells to resist apoptotic stimuli. This is due to differential modulation of the PKB/Bad pathway and of Bcl-X(L) expression. Thus, subtype-selective targeting of SKs will be critical when considering these enzymes as therapeutic targets for the treatment of inflammation or cancer.

Effects of eccentric cycle ergometry in alpine skiers

Eccentric cycling, where the goal is to resist the pedals, which are driven by a motor, increases muscle strength and size in untrained subjects. We hypothesized that it could also be beneficial for athletes, particularly in alpine skiing, which involves predominantly eccentric contractions at longer muscle lengths. We investigated the effects of replacing part of regular weight training with eccentric cycling in junior male alpine skiers using a matched-pair design. Control subjects ( N=7) executed 1-h weight sessions 3 times per week, which included 4-5 sets of 4 leg exercises. The eccentric group ( N=8) performed only 3 sets, followed by continuous sessions on the eccentric ergometer for the remaining 20 min. After 6 weeks, lean thigh mass increased significantly only in the eccentric group. There was a groupxtime effect on squat-jump height favouring the eccentric group, which also experienced a 6.5% improvement in countermovement-jump height. The ability to finely modulate muscle force during variable eccentric cycling improved 50% (p=0.004) only in the eccentric group. Although eccentric cycling did not significantly enhance isometric leg strength, we believe it is beneficial for alpine skiers because it provides an efficient means for hypertrophy while closely mimicking the type of muscle actions encountered while skiing.

Sperm of colourful males are better protected against oxidative stress

Sperm cells are highly vulnerable to free radicals, and sperm quality and male fertility are critically affected by oxidative stress. Recently, sexual ornaments, particularly carotenoid-based colourful traits, have been proposed to depend on a male's capacity to resist oxidative stress, and thus to signal sperm quality. We conducted an experimental test of this hypothesis on great tits Parus major, in which adults are sexually dichromatic in carotenoid-based breast plumage. We report the first evidence that ornaments and sperm quality may be linked through oxidative stress. When experimentally subjected to oxidative stress resulting from increased workload, less colourful males suffered a greater reduction in sperm motility and swimming ability, and increased levels of sperm lipid peroxidation compared to more colourful males. Moreover, the level of sperm lipid peroxidation was negatively correlated with sperm quality. Finally, carotenoid supplementation increased sperm quality of less colourful males, suggesting that pale males are deficient in carotenoid antioxidants.

Last stop expulsion - The minority question and forced migration in East-Central Europe: 1918-49

This article deals with the European minorities in the period between the two world wars and with their final expulsion from nation-states at the end of World War II. First, the tensions which arose between the organised minorities and the successor states of the Habsburg Monarchy are accounted for primarily by the argument that the various minorities located within the successor states had already undergone a comprehensive processes of nationalisation within the Habsburg Empire. Therefore they were able to resist assimilation by the political elites of the new titular nations (Czechs, Poles, Rumanians, Serbs). A second topic is that of the use made of the minorities issue by Adolf Hitler to help achieve his expansionist aims. The minorities issue was central to the international destabilisation of interwar Europe. Finally, the mass expulsion of minorities (above all, Germans) after the end of the war is explained by strategic considerations on the part of the Allied powers as well as involving the nation-state regimes. It is argued, against a commonly held view, that German atrocities during the period of occupation had little to do with the decision to expel most ethnic Germans from their territories of settlement in Poland, Czechoslovakia and Yugoslavia. The article shows that it is necessary to treat national minorities in the first half of the twentieth century as a single phenomenon which shares similar features across the various nation-states of East-Central Europe.

Thymic peptides for treatment of cancer patients

Background Purified thymus extracts (pTE) and synthetic thymic peptides (sTP) are thought to enhance the immune system of cancer patients in order to fight the growth of tumour cells and to resist infections due to immunosuppression induced by the disease and antineoplastic therapy. Objectives To evaluate the effectiveness of pTE and sTP for the management of cancer. Search methods We searched CENTRAL (The Cochrane Library 2010, Issue 3), MEDLINE, EMBASE, AMED, BIOETHICSLINE, BIOSIS, CATLINE, CISCOM, HEALTHSTAR, HTA, SOMED and LILACS (to February 2010). Selection criteria Randomised trials of pTE or sTP in addition to chemotherapy or radiotherapy, or both, compared to the same regimen with placebo or no additional treatment in adult cancer patients. Data collection and analysis Two authors independently extracted data from published trials. We derived odds ratios (OR) from overall survival (OS) and disease-free survival (DFS) rates, tumour response (TR) rates, and rates of adverse effects (AE) related to antineoplastic treatments. We used a random-effects model for meta-analysis. Main results We identified 26 trials (2736 patients). Twenty trials investigated pTE (thymostimulin or thymosin fraction 5) and six trials investigated sTP (thymopentin or thymosin α1). Twenty-one trials reported results for OS, six for DFS, 14 for TR, nine for AE and 10 for safety of pTE and sTP. Addition of pTE conferred no benefit on OS (RR 1.00, 95% CI 0.79 to 1.25); DFS (RR 0.97, 95% CI 0.82 to 1.16); or TR (RR 1.07, 95% CI 0.92 to 1.25). Heterogeneity was moderate to high for all these outcomes. For thymosin α1 the pooled RR for OS was 1.21 (95% CI 0.94 to 1.56, P = 0.14), with low heterogeneity; and 3.37 (95% CI 0.66 to 17.30, P = 0.15) for DFS, with moderate heterogeneity. The pTE reduced the risk of severe infectious complications (RR 0.54, 95% CI 0.38 to 0.78, P = 0.0008; I² = 0%). The RR for severe neutropenia in patients treated with thymostimulin was 0.55 (95% CI 0.25 to 1.23, P = 0.15). Tolerability of pTE and sTP was good. Most of the trials had at least a moderate risk of bias. Authors' conclusions Overall, we found neither evidence that the addition of pTE to antineoplastic treatment reduced the risk of death or disease progression nor that it improved the rate of tumour responses to antineoplastic treatment. For thymosin α1, there was a trend for a reduced risk of dying and of improved DFS. There was preliminary evidence that pTE lowered the risk of severe infectious complications in patients undergoing chemotherapy or radiotherapy.

Comparative stability studies of poly(2-methyl-2-oxazoline) and poly(ethylene glycol) brush coatings

Non-fouling surfaces that resist non-specific adsorption of proteins, bacteria, and higher organisms are of particular interest in diverse applications ranging from marine coatings to diagnostic devices and biomedical implants. Poly(ethylene glycol) (PEG) is the most frequently used polymer to impart surfaces with such non-fouling properties. Nevertheless, limitations in PEG stability have stimulated research on alternative polymers that are potentially more stable than PEG. Among them, we previously investigated poly(2-methyl-2-oxazoline) (PMOXA), a peptidomimetic polymer, and found that PMOXA shows excellent anti-fouling properties. Here, we compare the stability of films self-assembled from graft copolymers exposing a dense brush layer of PEG and PMOXA side chains, respectively, in physiological and oxidative media. Before media exposure both film types prevented the adsorption of full serum proteins to below the detection limit of optical waveguide in situ measurements. Before and after media exposure for up to 2 weeks, the total film thickness, chemical composition, and total adsorbed mass of the films were quantified using variable angle spectroscopic ellipsometry (VASE), X-ray photoelectron spectroscopy (XPS), and optical waveguide lightmode spectroscopy (OWLS), respectively. We found (i) that PMOXA graft copolymer films were significantly more stable than PEG graft copolymer films and kept their protein-repellent properties under all investigated conditions and (ii) that film degradation was due to side chain degradation rather than due to copolymer desorption.

The genomic Echinococcus microsatellite EmsB sequences: from a molecular marker to the epidemiological tool

In the field of molecular and epidemiological parasitology, characterization of fast evolving genetic markers appears as an important challenge to consider the diversity and genetic structure of parasites. The study of respective populations can help us to understand their adaptive strategies to survive and perpetuate the species within different host populations, all trying to resist infection. In the past, the relative monomorphic features of Echinococcus multilocularis, the causative agent of alveolar echinococcosis and a severe human parasitic disease, did not stimulate studies dealing with the genetic variability of Echinococcus species or respective populations. A recently developed, characterized and validated original multilocus microsatellite, named EmsB, tandemly repeated in the genome, offered an additional opportunity for this line of investigation. We have compiled in this review new insights brought by this molecular tracker on the transmission activity of Echinococcus among different hosts and at different geographical scales.

Fruit yogurt: any erosive potential regarding teeth?

OBJECTIVE: The capability of drinks and foods to resist pH changes brought about by salivary buffering may play an important role in the dental erosion process in children. The aim of the present study was to test fruit yogurt, a popular snack for children, and the degrees of saturation (pK-pl) with respect to hydroxyapatite and fluorapatite to determine their erosive potential. METHOD AND MATERIALS: A variety of fruit yogurt was tested. To test the pH, 8 readings were taken with a pH electrode for each yogurt. Calcium content was detected by atomic absorption spectrophotometer, phosphorus by the inductively coupled plasma method, and fluoride content by ion chromatography. The degrees of saturation of hydroxyapatite and fluorapatite were calculated by use of a computer program. Statistical analysis was performed using 2-tailed analysis of variance (P < .05) and a post hoc test (Tukey) to determine differences between groups. RESULTS: The pH of each fruit concentrate was significantly different, except for banana yogurt. Except for the phosphorus content of raspberry yogurt, the calcium and phosphorus content for each fruit concentrate were significantly different. Fluoride levels were the same for all yogurts tested, and the degrees of saturation of hydroxyapatite and fluorapatite was positive, indicating supersaturation. CONCLUSION: It could be stated that fruit yogurt has no erosive potential.

Erosive and buffering capacities of yogurt

OBJECTIVE: The capability of drinks and foods to resist pH changes brought about by salivary buffering may play an important role in the erosion of dental enamel. The aim of the present study was to measure the initial pH of several types of yogurt and to test the degrees of saturation (pK-pl) with respect to hydroxyapatite and fluorapatite to determine the buffering capacity and related erosive potential of yogurt. METHOD AND MATERIALS: Twenty-five milliliters of 7 types of freshly opened yogurt was titrated with 1 mol/L of sodium hydroxide, added in 0.5 mL increments, until the pH reached 10, to assess the total titratable acidity, a measure of the drink's own buffering capacity. The degrees of saturation (pK-pl) with respect to hydroxyapatite and fluorapatite were also calculated, using a computer program developed for this purpose. For statistical analysis, samples were compared using Kruskal-Wallis test. RESULTS: The buffering capacities can be ordered as follows: fruit yogurt >low-fat yogurt >bioyogurt >butter yogurt >natural yogurt >light fruit yogurt >light yogurt. The results suggest that, in vitro, fruit yogurt has the greatest buffering capacity. CONCLUSION: It can be stated that it is not possible to induce erosion on enamel with any type of yogurt.

Characterization of a stem cell population in lung cancer A549 cells

We isolated a stem cell subpopulation from human lung cancer A549 cells using FACS/Hoechst 33342. This side population (SP), which comprised 24% of the total cell population, totally disappeared after treatment with the selective ABCG 2 inhibitor fumitremorgin C. In a repopulation study, isolated SP and non-SP cells were each able to generate a heterogeneous population of SP and non-SP cells, but this repopulation occurred more rapidly in SP cells than non-SP. An MTT assay and cell cycle distribution analysis reveal a similar profile between SP and non-SP groups. However, in the presence of doxorubicin (DOX) and methotrexate (MTX), SP cells showed significantly lower Annexin V staining when compared to non-SP cells. Taken together, these results demonstrate that SP cells have an active regeneration capacity and high anti-apoptotic activity compared with non-SP cells. Furthermore, our GeneChip data revealed a heightened mRNA expression of ABCG2 and ABCC2 in SP cells. Overall these data explain why the SP of A549 has a unique ability to resist DOX and MTX treatments. Therefore, we suggest that the expression of the ABCG2 transporter plays an important role in the multidrug resistance phenotype of A549 SP cells.

Chondrocyte mechanotransduction: effects of compression on deformation of intracellular organelles and relevance to cellular biosynthesis

OBJECTIVE: The effects of mechanical deformation of intact cartilage tissue on chondrocyte biosynthesis in situ have been well documented, but the mechanotransduction pathways that regulate such phenomena have not been elucidated completely. The goal of this study was to examine the effects of tissue deformation on the morphology of a range of intracellular organelles which play a major role in cell biosynthesis and metabolism. DESIGN: Using chemical fixation, high pressure freezing, and electron microscopy, we imaged chondrocytes within mechanically compressed cartilage explants at high magnification and quantitatively and qualitatively assessed changes in organelle volume and shape caused by graded levels of loading. RESULTS: Compression of the tissue caused a concomitant reduction in the volume of the extracellular matrix (ECM), chondrocyte, nucleus, rough endoplasmic reticulum, and mitochondria. Interestingly, however, the Golgi apparatus was able to resist loss of intraorganelle water and retain a portion of its volume relative to the remainder of the cell. These combined results suggest that a balance between intracellular mechanical and osmotic gradients govern the changes in shape and volume of the organelles as the tissue is compressed. CONCLUSIONS: Our results lead to the interpretive hypothesis that organelle volume changes appear to be driven mainly by osmotic interactions while shape changes are mediated by structural factors, such as cytoskeletal interactions that may be linked to extracellular matrix deformations. The observed volume and shape changes of the chondrocyte organelles and the differential behavior between organelles during tissue compression provide evidence for an important mechanotransduction pathway linking translational and post-translational events (e.g., elongation and sulfation of glycosaminoglycans (GAGs) in the Golgi) to cell deformation.

Immunotoxic effects of environmental toxicants in fish - how to assess them?

Numerous environmental chemicals, both long-known toxicants such as persistent organic pollutants as well as emerging contaminants such as pharmaceuticals, are known to modulate immune parameters of wildlife species, what can have adverse consequences for the fitness of individuals including their capability to resist pathogen infections. Despite frequent field observations of impaired immunocompetence and increased disease incidence in contaminant-exposed wildlife populations, the potential relevance of immunotoxic effects for the ecological impact of chemicals is rarely considered in ecotoxicological risk assessment. A limiting factor in the assessment of immunotoxic effects might be the complexity of the immune system what makes it difficult (1) to select appropriate exposure and effect parameters out of the many immune parameters which could be measured, and (2) to evaluate the significance of the selected parameters for the overall fitness and immunocompetence of the organism. Here, we present - on the example of teleost fishes - a brief discussion of how to assess chemical impact on the immune system using parameters at different levels of complexity and integration: immune mediators, humoral immune effectors, cellular immune defenses, macroscopical and microscopical responses of lymphoid tissues and organs, and host resistance to pathogens. Importantly, adverse effects of chemicals on immunocompetence may be detectable only after immune system activation, e.g., after pathogen challenge, but not in the resting immune system of non-infected fish. Current limitations to further development and implementation of immunotoxicity assays and parameters in ecotoxicological risk assessment are not primarily due to technological constraints, but are related from insufficient knowledge of (1) possible modes of action in the immune system, (2) the importance of intra- and inter-species immune system variability for the response against chemical stressors, and (3) deficits in conceptual and mechanistic assessment of combination effects of chemicals and pathogens.

Vibrations, Posture, and the Stabilization of Gaze: An Experimental Study on Impedance Control

Vibrations, Posture, and the Stabilization of Gaze: An Experimental Study on Impedance Control R. KREDEL, A. GRIMM & E.-J. HOSSNER University of Bern, Switzerland Introduction Franklin and Wolpert (2011) identify impedance control, i.e., the competence to resist changes in position, velocity or acceleration caused by environmental disturbances, as one of five computational mechanisms which allow for skilled and fluent sen-sorimotor behavior. Accordingly, impedance control is of particular interest in situa-tions in which the motor task exhibits unpredictable components as it is the case in downhill biking or downhill skiing. In an experimental study, the question is asked whether impedance control, beyond its benefits for motor control, also helps to stabi-lize gaze what, in turn, may be essential for maintaining other control mechanisms (e.g., the internal modeling of future states) in an optimal range. Method In a 3x2x4 within-subject ANOVA design, 72 participants conducted three tests on visual acuity and contrast (Landolt / Grating and Vernier) in two different postures (standing vs. squat) on a platform vibrating at four different frequencies (ZEPTOR; 0 Hz, 4 Hz, 8 Hz, 12 Hz; no random noise; constant amplitude) in a counterbalanced or-der with 1-minute breaks in-between. In addition, perceived exertion (Borg) was rated by participants after each condition. Results For Landolt and Grating, significant main effects for posture and frequency are re-vealed, representing lower acuity/contrast thresholds for standing and for higher fre-quencies in general, as well as a significant interaction (p < .05), standing for in-creasing posture differences with increasing frequencies. Overall, performance could be maintained at the 0 Hz/standing level up to a frequency of 8 Hz, if bending of the knees was allowed. The fact that this result is not only due to exertion is proved by the Borg ratings showing significant main effects only, i.e., higher exertion scores for standing and for higher frequencies, but no significant interaction (p > .40). The same pattern, although not significant, is revealed for the Vernier test. Discussion Apparently, postures improving impedance control not only turn out to help to resist disturbances but also assist in stabilizing gaze in spite of these perturbations. Con-sequently, studying the interaction of these control mechanisms in complex unpre-dictable environments seems to be a fruitful field of research for the future. References Franklin, D. W., & Wolpert, D. M. (2011). Computational mechanisms of sensorimotor control. Neuron, 72, 425-442.