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Key Clinical Message A melanoma of the upper eyelid was resected in a gray warmblood gelding. A full functional eyelid could be obtained by completion a sliding skin graft with a free labial mucocutaneous graft transplantation to restore the mucocutaneous junction and to decrease the risk of postoperative trichiasis.

Abstract Background CD4+ T cells are of great importance in the pathogenesis of systemic lupus erythematosus (SLE), as an imbalance between CD4+ regulatory T cells (Tregs) and CD4+ responder T cells (Tresps) causes flares of active disease in SLE patients. In this study, we aimed to find the role of aberrant Treg/Tresp cell differentiation for maintaining Treg/Tresp cell balance and Treg functionality. Methods To determine differences in the differentiation of Tregs/Tresps we calculated the percentages of CD45RA+CD31+ recent thymic emigrant (RTE) Tregs/Tresps and CD45RA+CD31− mature naive (MN) Tregs/Tresps, as well as CD45RA−CD31+ and CD45RA−CD31− memory Tregs/Tresps (CD31+ and CD31− memory Tregs/Tresps) within the total Treg/Tresp pool of 78 SLE remission patients compared with 94 healthy controls of different ages. The proliferation capacity of each Treg/Tresp subset was determined by staining the cells with anti-Ki67 monoclonal antibodies. Differences in the autologous or allogeneic Treg function between SLE remission patients and healthy controls were determined using suppression assays. Results With age, we found an increased differentiation of RTE Tregs via CD31+ memory Tregs and of RTE Tresps via MN Tresps into CD31− memory Tregs/Tresp in healthy volunteers. This opposite differentiation of RTE Tregs and Tresps was associated with an age-dependent increase in the suppressive activity of both naive and memory Tregs. SLE patients showed similar age-dependent Treg cell differentiation. However, in these patients RTE Tresps differentiated increasingly via CD31+ memory Tresps, whereby CD31− memory Tresps arose that were much more difficult to inhibit for Tregs than those that emerged through differentiation via MN Tresps. Consequently, the increase in the suppressive activity of Tregs with age could not be maintained in SLE patients. Testing the Tregs of healthy volunteers and SLE patients with autologous and nonautologous Tresps revealed that the significantly decreased Treg function in SLE patients was not exclusively attributed to an age-dependent diminished sensitivity of the Tresps for Treg suppression. The immunosuppressive therapy reduced the accelerated age-dependent Tresp cell proliferation to normal levels, but simultaneously inhibited Treg cell proliferation below normal levels. Conclusions Our data reveal that the currently used immunosuppressive therapy has a favorable effect on the differentiation and proliferation of Tresps but has a rather unfavorable effect on the proliferation of Tregs. Newer substances with more specific effects on the immune system would be desirable.

Abstract Background Gluconobacter oxydans is a strictly aerobic Gram-negative acetic acid bacterium used industrially for oxidative biotransformations due to its exceptional type of catabolism. It incompletely oxidizes a wide variety of carbohydrates regio- and stereoselectively in the periplasm using membrane-bound dehydrogenases with accumulation of the products in the medium. As a consequence, only a small fraction of the carbon and energy source enters the cell, resulting in a low biomass yield. Additionally, central carbon metabolism is characterized by the absence of a functional glycolysis and absence of a functional tricarboxylic acid (TCA) cycle. Due to these features, G. oxydans is a highly interesting model organism. Here we analyzed global mRNA decay in G. oxydans to describe its characteristic features and to identify short-lived mRNAs representing potential bottlenecks in the metabolism for further growth improvement by metabolic engineering. Results Using DNA microarrays we estimated the mRNA half-lives in G. oxydans. Overall, the mRNA half-lives ranged mainly from 3 min to 25 min with a global mean of 5.7 min. The transcripts encoding GroES and GroEL required for proper protein folding ranked at the top among transcripts exhibiting both long half-lives and high abundance. The F-type H+-ATP synthase transcripts involved in energy metabolism ranked among the transcripts with the shortest mRNA half-lives. RNAseq analysis revealed low expression levels for genes of the incomplete TCA cycle and also the mRNA half-lives of several of those were short and below the global mean. The mRNA decay analysis also revealed an apparent instability of full-length 23S rRNA. Further analysis of the ribosome-associated rRNA revealed a 23S rRNA fragmentation pattern exhibiting new cleavage regions in 23S rRNAs which were previously not known. Conclusions The very short mRNA half-lives of the H+-ATP synthase, which is likely responsible for the ATP-proton motive force interconversion in G. oxydans under many or most conditions, is notably in contrast to mRNA decay data from other bacteria. Together with the short mRNA half-lives and low expression of some other central metabolic genes it could limit intended improvements of G. oxydans’ biomass yield by metabolic engineering. Also, further studies are needed to unravel the multistep process of the 23S rRNA fragmentation in G. oxydans.

Abstract Background When evaluating the toxicity of engineered nanomaterials (ENMS) it is important to use multiple bioassays based on different mechanisms of action. In this regard we evaluated the use of gene expression and common cytotoxicity measurements using as test materials, two selected nanoparticles with known differences in toxicity, 5 nm mercaptoundecanoic acid (MUA)-capped InP and CdSe quantum dots (QDs). We tested the effects of these QDs at concentrations ranging from 0.5 to 160 µg/mL on cultured normal human bronchial epithelial (NHBE) cells using four common cytotoxicity assays: the dichlorofluorescein assay for reactive oxygen species (ROS), the lactate dehydrogenase assay for membrane viability (LDH), the mitochondrial dehydrogenase assay for mitochondrial function, and the Comet assay for DNA strand breaks. Results The cytotoxicity assays showed similar trends when exposed to nanoparticles for 24 h at 80 µg/mL with a threefold increase in ROS with exposure to CdSe QDs compared to an insignificant change in ROS levels after exposure to InP QDs, a twofold increase in the LDH necrosis assay in NHBE cells with exposure to CdSe QDs compared to a 50% decrease for InP QDs, a 60% decrease in the mitochondrial function assay upon exposure to CdSe QDs compared to a minimal increase in the case of InP and significant DNA strand breaks after exposure to CdSe QDs compared to no significant DNA strand breaks with InP. High-throughput quantitative real-time polymerase chain reaction (qRT-PCR) data for cells exposed for 6 h at a concentration of 80 µg/mL were consistent with the cytotoxicity assays showing major differences in DNA damage, DNA repair and mitochondrial function gene regulatory responses to the CdSe and InP QDs. The BRCA2, CYP1A1, CYP1B1, CDK1, SFN and VEGFA genes were observed to be upregulated specifically from increased CdSe exposure and suggests their possible utility as biomarkers for toxicity. Conclusions This study can serve as a model for comparing traditional cytotoxicity assays and gene expression measurements and to determine candidate biomarkers for assessing the biocompatibility of ENMs.

For a secure integration of weather dependent renewable energies in Germany's mixed power supply, precise forecasts of expected wind power are indispensable. These in turn are heavily dependent on numerical weather prediction (NWP). With this relevant area of application, NWP models need to be evaluated concerning new variables such as wind speed at hub heights of wind power plants. This article presents verification results of the deterministic NWP forecasts of the global ICON model, its ICON-EU nest, the COSMO-EU, and the COSMO-DE as well as of the ensemble prediction system COSMO-DE-EPS of the German National Weather Service (DWD), against wind mast observations. The focus is on the diurnal cycle in the Planetary Boundary Layer as wind power forecasts for Germany exhibit pronounced systematic amplitude and phase errors in the morning and evening hours. NWP forecasts with lead times up to 48 hours are examined. All considered NWP models reveal shortcomings concerning the representation of the diurnal cycle. Especially in summertime at onshore locations, when Low-Level Jets form, nocturnal wind speeds at hub height are underestimated. In the COSMO model, stable conditions are not sufficiently reflected in the first part of the night and the vertical mixing after sunrise establishes too late. The verification results of the COSMO-DE-EPS confirm the deficiencies of the deterministic forecasts. The deficiencies are present in all ensemble members and thus indicate potential for improvement not only in the model physics parameterization but also concerning the physical ensemble perturbations.

With the strong expansion of the installed renewable energy over the last years, the relevance of weather forecasts for operating the German power system has considerably increased. In that context, rare but important events like the solar eclipse on the morning of 20 March 2015 pose an additional challenge when operating the power system, as it affects the photovoltaic (PV) power production by inducing strong gradients in the feed-in. In order to maintain grid stability, the uncertainties associated with the eclipse have been estimated in advance for planning necessary precautions. Especially the maximum gradients in PV-power were of importance for the provision of balancing energy. Numerical weather prediction (NWP) is very suited for this assessment, as it allows to consider the complex mechanisms occurring in the atmosphere. Thus the impact of the eclipse on meteorological parameters which affect the PV-power generation were evaluated. Sensitivity studies with NWP models have been conducted in order to assess the reduction in short wave radiation and temperature during the total solar eclipse months before the actual event. For this purpose, model simulations with the non-hydrostatic COSMO models from the German Weather Service (DWD) have been performed over Germany and Europe. As the weather situation and especially the cloud cover during the eclipse could not be known in advance, a realistic worst case (clear sky conditions) and a best case (overcast conditions) scenario were simulated over Germany. Thereof the PV-power production has been estimated and analyzed for the different scenarios. The NWP model data from the sensitivity studies are openly distributed (doi:10.1594/PANGAEA.839163). As near real-time NWP simulations considering the solar eclipse were conducted a few days prior to the event, they are herein validated with measurements. Furthermore, the actual PV-power production and actions taken by the TSOs during the solar eclipse are stated.

As enteroparasitoses são infecções causadas por parasitas intestinais que constituem um dos grandes problemas de saúde pública em todo o mundo, sofrendo variações quanto à região de cada país e tendo sua maior prevalência vinculada a áreas que se apresentam com condições higiênico-sanitárias precárias. O objetivo deste estudo foi verificar a prevalência de enteroparasitoses em crianças de creches da rede pública e privada de Santo Ângelo-RS. Foram coletadas 51 amostras de fezes de escolares com idades entre 1 e 3 anos, no período de março a maio de 2016, utilizando-se o método de Ritchie para análise. Além disso, foram verificados através de um questionário fatores socioeconômicos e aspectos relacionados à higiene, caracterizando um estudo observacional, transversal e prospectivo. Das 51 amostras analisadas, foi registrada uma prevalência de parasitoses de 18% sendo 16% referentes às creches públicas e 2% as creches particulares. Os parasitas encontrados foram Endolimax nana (67%), Giardia lamblia (22%) e Trichuris trichiura (11%). As crianças do sexo masculino apresentaram uma frequência de parasitismo de 78%, o que diferiu da frequência observada entre as crianças do sexo feminino que foi de 22%. Os resultados indicam valores menores que os encontrados na literatura, entretanto o risco de contaminação e incidência de novos casos está presente na comunidade. Portanto, há necessidade da implantação de práticas educacionais que possam instruir a população para a prevenção de parasitoses e para a conscientização da população, principalmente em relação à educação sanitária, além do mais, o exame parasitológico de fezes pode ser realizado rotineiramente para controle dessas infecções.

Recent studies show that regulatory T cells (Tregs) play an essential role in tolerance induction after organ transplantation. In order to examine whether there are differences in the composition of the total CD4(+)CD127(low+/-)FoxP3(+)- Treg cell pool between stable transplant patients and patients with biopsy proven rejection (BPR), we compared the percentages and the functional activity of the different Treg cell subsets (DR(high+)CD45RA(-)-Tregs, DR(low+)CD45RA(-)-Tregs, DR(-)CD45RA(-)-Tregs, DR(-)CD45RA(+)-Tregs). All parameters were determined during the three different periods of time after transplantation (0-30 days, 31-1,000 days, >1,000 days). Among 156 transplant patients, 37 patients suffered from BPR. The most prominent differences between rejecting and non-rejecting patients were observed regarding the DR(high+)CD45RA(-)-Treg cell subset. Our data demonstrate that the suppressive activity of the total Treg pool strongly depends on the presence of these Treg cells. Their percentage within the total Treg pool strongly decreased after transplantation and remained relatively low during the first year after transplantation in all patients. Subsequently, the proportion of this Treg subset increased again in patients who accepted the transplant and reached a value of healthy non-transplanted subjects. By contrast, in patients with acute kidney rejection, the DR(high+)CD45RA(-)-Treg subset disappeared excessively, causing a reduction in the suppressive activity of the total Treg pool. Therefore, both the monitoring of its percentage within the total Treg pool and the monitoring of the HLA-DR MFI of the DR(+)CD45RA(-)-Treg subset may be useful tools for the prediction of graft rejection.

Isothermal microcalorimetry is an established tool to measure heat flow of physical, chemical or biological processes. The metabolism of viable cells produces heat, and if sufficient cells are present, their heat production can be assessed by this method. In this study, we investigated the heat flow of two medically important protozoans, Trypanosoma brucei rhodesiense and Plasmodium falciparum. Heat flow signals obtained for these pathogens allowed us to monitor parasite growth on a real-time basis as the signals correlated with the number of viable cells. To showcase the potential of microcalorimetry for measuring drug action on pathogenic organisms, we tested the method with three antitrypanosomal drugs, melarsoprol, suramin and pentamidine and three antiplasmodial drugs, chloroquine, artemether and dihydroartemisinin, each at two concentrations on the respective parasite. With the real time measurement, inhibition was observed immediately by a reduced heat flow compared to that in untreated control samples. The onset of drug action, the degree of inhibition and the time to death of the parasite culture could conveniently be monitored over several days. Microcalorimetry is a valuable element to be added to the toolbox for drug discovery for protozoal diseases such as human African trypanosomiasis and malaria. The method could probably be adapted to other protozoan parasites, especially those growing extracellularly.

Behavioral endophenotypes are determined by a multitude of counteracting but precisely balanced molecular and physiological mechanisms. In this study, we aim to identify potential novel molecular targets that contribute to the multigenic trait "anxiety". We used microarrays to investigate the gene expression profiles of different brain regions within the limbic system of mice which were selectively bred for either high (HAB) or low (LAB) anxiety-related behavior, and also show signs of comorbid depression-like behavior. We identified and confirmed sex-independent differences in the basal expression of 13 candidate genes, using tissue from the entire brain, including coronin 7 (Coro7), cathepsin B (Ctsb), muscleblind-like 1 (Mbnl1), metallothionein 1 (Mt1), solute carrier family 25 member 17 (Slc25a17), tribbles homolog 2 (Trib2), zinc finger protein 672 (Zfp672), syntaxin 3 (Stx3), ATP-binding cassette, sub-family A member 2 (Abca2), ectonucleotide pyrophosphatase/phosphodiesterase 5 (Enpp5), high mobility group nucleosomal binding domain 3 (Hmgn3) and pyruvate dehydrogenase beta (Pdhb). Additionally, we confirmed brain region-specific differences in the expression of synaptotagmin 4 (Syt4).Our identification of about 90 polymorphisms in Ctsb suggested that this gene might play a critical role in shaping our mouse model's behavioral endophenotypes. Indeed, the assessment of anxiety-related and depression-like behaviors of Ctsb knock-out mice revealed an increase in depression-like behavior in females. Altogether, our results suggest that Ctsb has significant effects on emotionality, irrespective of the tested mouse strain, making it a promising target for future pharmacotherapy.