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IntroductionImmunosuppressive therapy prevents graft rejection but increases the risk of non-melanoma skin cancer (NMSC), especially in elderly kidney transplant recipients (KTR).MethodsIn this study, we separately investigated the differentiation of CD8+ regulatory T cells (Tregs) and responder T cells (Tresps) between healthy KTR without NMSC, KTR developing de-novo NMSC within two years after the enrolment, and KTR with NMSC at the time of enrolment. Antigen-unexperienced CCR7+CD45RA+CD31+ recent thymic emigrant (RTE) cells differentiate via CD45RA-CD31+ memory (CD31+ memory) cells, via resting mature naïve (MN) cells or via direct proliferation into CD45RA-CD31- memory (CD31- memory) cells, consisting of both CCR7+CD45RA- central memory (CM) and CCR7-CD45RA- effector memory (EM) cells.ResultsWe found that both RTE Treg and Tresp differentiation via CD31+ memory Tregs/Tresps was age-independently increased in KTR, who developed de novo NMSC during the follow-up period, causing abundant CM Treg/Tresp production, which may be crucial for cancer immunity. These changes favored a strongly increased CD8+ Treg/Tresp ratio, suggesting this ratio as a reliable marker for de-novo NMSC development in KTR. However, with age, this differentiation was replaced by increased conversion of resting MN Tregs/Tresps into CM Tregs/Tresps, which exhausted for Tresps but not for Tregs. In KTR with already existing NMSC at enrolment, differentiation was maintained via conversion and proliferation of resting MN Tregs/Tresps, which however increasingly exhausted with age, especially for Tresps. This resulted in a strong accumulation of terminally differentiated effector memory (TEMRA) Tresps in elderly individuals. Patients with NMSC recurrence showed increased proliferation of resting MN Tregs/Tresps into EM Tregs/Tresps, which tended to exhaust more rapidly, particularly for Tresps, than in patients without NMSC recurrence.DiscussionIn conclusion, we provide evidence that immunosuppressive therapy inhibits differentiation of CD8+ Tregs more than that of CD8+ Tresps, resulting in an exhausted Tresp profile, thus providing a possible therapeutic approach to improve poor cancer immunity in elderly KTR.

To confirm discrepant SARS-CoV-2-IgG results in four standard assays we applied for the first time a prototype of a coronavirus IgG-line-blot which employs antigens from seasonal coronaviruses, SARS-1 and SARS-CoV-2 combined with avidity testing as a confirmatory tool in a follow-up of five cases including pre-pandemic samples.

Microbial synthesis of monolignols and lignans from simple substrates is a promising alternative to plant extraction. Bottlenecks and byproduct formation during heterologous production require targeted metabolomics tools for pathway optimization.In contrast to available fractional methods, we established a comprehensive targeted metabolomics method. It enables the quantification of 17 extra- and intracellular metabolites of the monolignol and lignan pathway, ranging from amino acids to pluviatolide. Several cell disruption methods were compared. Hot water extraction was best suited regarding monolignol and lignan stability as well as extraction efficacy. The method was applied to compare enzymes for alleviating bottlenecks during heterologous monolignol and lignan production in E. coli. Variants of tyrosine ammonia-lyase had a considerable influence on titers of subsequent metabolites. The choice of multicopper oxidase greatly affected the accumulation of lignans. Metabolite titers were monitored during batch fermentation of either monolignol or lignan-producing recombinant E. coli strains, demonstrating the dynamic accumulation of metabolites.The new method enables efficient time-resolved targeted metabolomics of monolignol- and lignan-producing E. coli. It facilitates bottleneck identification and byproduct quantification, making it a valuable tool for further pathway engineering studies. This method will benefit the bioprocess development of biotransformation or fermentation approaches for microbial lignan production.

During the COVID-19 pandemic, healthcare workers have a high workload and have been exposed to various psychosocial stressors. This study aimed to evaluate health workers during the COVID-19 pandemic in the world. The method used in this research is qualitative with a literature review approach. The data sources in this study were taken from the Scopus database using the keywords “health workers,” “burnout,” AND “coronavirus” from the time range of 2020 to April 25, 2022. From the determination of the time range, 150 documents emerged. This study revealed that the Pearson correlation between total burnout scale scores on healthcare workers, professionals, exhaustion, mental, stress, personal, depression, symptoms, emergency, system, job, and impact indicated that overall burnout scores were associated with depression and anxiety. Stress symptoms had correlation values ranging from 0.84 to 0.73. Job burnout had a significant relationship with exhaustion at 0.89; depression r = 0.73), impacting a score of 0.66. At the same time, the fulfillment of professional and interpersonal disengagement showed a Pearson correlation between the total burnout scale scores on health workers, professionals, exhaustion, mental, stress, personal, depression, symptoms, emergency, system, job, and impact. Overall, the participants in health care workers worldwide experienced high levels of psychological distress. We also found that health workers dealing with COVID-19 pandemic patients were more likely to experience depression, stress, and burnout than health staff who were not personally involved in medical work. Furthermore, this study will be a follow-up study using the Work Ability Index (WAI) to measure work ability and work satisfaction.

Abstract Treatment of deep corneal ulcers using small intestinal submucosa (SIS) without corneal sutures in combination with a third eyelid flap seems a promising technical modification. Advantages are a reduction in surgery time and no additional damage of healthy corneal tissue.

Abstract Background Brachycephalic breeds have anatomical skull changes that are responsible for ocular clinical signs, known as the brachycephalic ocular syndrome (BOS). Their popularity has increased in recent years but the excessive pressure of selection lead to extreme conformation of skull shapes, resulting in facial alterations that can put these dogs’ vision at risk. Objectives This study aimed to analyse the ocular disorders in a sample of 93 brachycephalic dogs to better characterize the disease complex BOS. Material and methods Brachycephalic dogs were submitted to a complete ophthalmological examination. The studied parameters included animal’s sex, age and breed, age, ophthalmological tests performed, results of complementary exams, clinical signs, ocular disorders, treatment protocols and their outcomes. Data were organized using Microsoft Office Excel 2007® and statistical analysis was performed with IBM SPSS Statistics 20®. Results The studied population included 93 brachycephalic dogs 45 males (48%) and 48 females (52%) from different breeds: French Bulldog (n = 38), Shih-Tzu (n = 22), Pug (n = 17), English Bulldog (n = 5), Pekingese (n = 4), Boxer (n = 4) and Boston Terrier (n = 3), aged between 0.2–16 years, median 4.65 years. The most frequent ocular abnormalities were corneal ulcers in 44%, corneal pigmentation in 36%, corneal fibrosis in 25% and entropion in 22% of the animals. There was a higher incidence of corneal pigmentary keratitis in Pugs (53%) and corneal fibrosis in Shih Tzus (36%). The most common surgical techniques were medial canthoplasty in 22%, conjunctival flap in 10% and electroepilation in 7% of the cases, without post-operative complications. Conclusions: This study contributed to a better characterization of the disease complex brachycephalic ocular syndrome. The percentage of ocular disorders like entropion, corneal pigmentation, fibrosis and ulcers was high, highlighting the importance of a regular ophthalmological check-up, and early diagnosis of the primary disorders. A higher incidence of corneal pigmentation was noticed in Pugs and corneal fibrosis in Shih Tzus, which suggests that some brachycephalic breeds may be predisposed to certain ocular abnormalities. A responsible reproductive strategy should be implemented to avoid undesired transmission of the abnormal traits to the offspring.

The annual influenza vaccination has been officially recommended for medical staff in Germany since 1988. Nevertheless, the vaccination rate among medical staff is still low. The present study deals with the influenza vaccination rate of staff at a German University hospital over time as well as with the reasons that led to a positive vaccination decision and the barriers to acceptance of vaccination. For this purpose, the staff members received questionnaires in which they were asked about influenza vaccination and the reasons for or against vaccination. In addition, the questionnaire contains information on gender, age group, occupational group and presence of a chronic co-morbidity. Logistic regression analysis was used to investigate which of these predictors most strongly influenced the vaccination decision. It was shown that the reasons for or against vaccination differ significantly between the occupational groups and that the occupational group affiliation has the greatest influence on the vaccination decision in the comparison of the investigated predictors. In order to achieve a positive influence on vaccination acceptance, future measures should focus on increasing confidence in vaccination and on increasing the perception of risk from influenza illness. The findings may contribute to future targeted strategies to increase vaccination rates and suggest occupational group-specific interventions.

Chronic kidney failure (KF) provokes the development of immune senescent CD8+ cytotoxic T cells, affecting the occurrence of graft rejection, viral infections, and malignancies after kidney transplantation. In this study, we analyzed the impact of KF, subsequent dialysis treatment, and kidney transplantation on the differentiation of CD8+CD31+CD45RA+CCR7+ recent thymic emigrant (CCR7+ RTE) Tregs/Tresps into CD8+CD31-CD45RA- memory (CD31- memory) Tregs/Tresps and its effect on the release of cytokines, Fas receptor, Fas ligand as well as cytotoxic mediators by naïve, central memory (CM), effector memory (EM), and terminally differentiated effector memory (TEMRA) Tresps. We found that normal age-dependent differentiation of CD8+ Tregs/Tresps generally differs in the way that TEMRA cells only arise in Tresps. Compared to healthy controls, KF patients revealed an age-independently decreased frequency of CCR7+ RTE Tregs/Tresps, but increased frequencies of CCR7+ MN Tregs/Tresps and CD31- memory Tregs/Tresps, suggesting an increased differentiation via CD31+CD45RA- memory (CD31+ memory) Tregs/Tresps into CD31- memory Tregs/Tresps. Intensified differentiation via CD31+ memory Tresps increased the emergence of apoptosis-resistant CM Tresps with strong Fas ligand-mediated cytotoxicity. CCR7+ RTE Tresp proliferation generated TEMRA Tresps, secreting high levels of cytotoxic mediators. In dialysis and transplant patients, CD31+ TEMRA Tregs/Tresps accumulated, proposing an impaired CCR7+ RTE Treg/Tresp differentiation via CD31+ memory Tregs/Tresps into CD31- memory Tregs/Tresps. Increased percentages of CD31- TEMRA Tresps, but not of CD31- TEMRA Tregs, were observed in all patient groups, indicating impaired proliferation of CCR7+ RTE Tresps, but not of CCR7+ RTE Tregs, into CD31- memory Tregs/Tresps. In transplant patients, CCR7+ RTE Tregs accumulated, while frequencies of CCR7+ RTE Tresps were decreased, suggesting that the immunosuppressive therapy only prevented excessive CCR7+ RTE Treg differentiation but not that of CCR7+ RTE Tresps. Presumably, this caused the accumulation of TEMRA Tresps with decreased release of cytotoxic mediators, such as perforin. In conclusion, we propose that chronic KF affects both the differentiation of CD8+ Tregs and CD8+ Tresps. However, the immunosuppressive therapy after transplantation may successfully prevent excessive Treg differentiation, but not as suffciently that of Tresps. Therefore, the risk for graft rejection may be reduced, while the susceptibility for infections and malignancies may be increased in these patients.

O artigo visa responder à pergunta: o que explica a ausência de políticas de gênero nos documentos oficiais da defesa brasileira? Para tanto, sugere que tal ausência é resultado, também, da influência militar na política. Isto é, os desafios das mulheres militares brasileiras, como o acesso limitado a certas armas e especialidades, restrições nas formas de ingresso, problemas de infraestrutura e inexistência de uma legislação que as ampare, são reflexos dos interesses das elites políticas e sua relação com os líderes militares, os quais nem sempre possuem vontade política para introduzir a questão de gênero na agenda, esforçando-se para manter a organização ilesa às mudanças sociais. O artigo emprega a técnica de rastreamento de processo, com especial enfoque na busca de evidências empíricas.

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.