Role of monocyte-induced development of Th17 cells, the heat shock protein 90 and proinflammatory S100 proteins in the pathogenesis of graft-versus-host disease

Allogeneic hematopoietic cell transplantation (HCT) is an effective treatment for patients with hematologic malignancies, aplastic anaemia, and congenital immunodeficiency disorders. One of the major serious morbidities associated with HCT is the development of acute or chronic graft-versus-host disease (GvHD). The pathophysiology of GvHD is complex and not fully understood. The role of Th17 cells during GvHD is discussed controversially and still remains unclear. In this study, the induction of Th17 cells by monocytes of patients with GvHD in vitro was analysed demonstrating that monocytes isolated from patients with acute skin and intestinal GvHD stage I-IV or chronic GvHD induce significantly increased levels of Th17 cells compared to patients without GvHD after HCT and healthy controls. Several studies suggest using the determined levels of regulatory (Treg) cells and the ratio of Th17 cells to Treg cells in the peripheral blood of patients as diagnostic markers for GvHD. However, the data of the present study have demonstrated that the determined percentages of Treg cells in peripheral blood mononuclear cells (PBMCs) isolated from patients with acute GvHD do not differ from the assessed percentages of Treg cells in PBMCs of healthy donors and patients without GvHD after HCT. By contrast, the percentages of Treg cells in PBMCs from patients with extensive chronic GvHD seem to be increased in comparison to the healthy controls and the non-GvHD group. The results of the present work further indicate that the calculated ratios of Th17 cells to Treg cells are not altered in patients with acute or chronic GvHD compared to patients without GvHD after HCT. Development and progression of GvHD is mediated by multiple cellular and inflammatory effectors. However, several of these molecules are still unknown. Previous studies have demonstrated that S100 proteins act as innate amplifier of inflammation and play an important role in many inflammatory diseases such as inflammatory bowel disease or rheumatoid arthritis. These proinflammatory S100 proteins belong to the group of Damage Associated Molecular Pattern (DAMP) molecules and are released by activated or damaged phagocytes under conditions of cell stress during infections and autoimmune diseases. Therefore, expression levels of S100 proteins in monocytes and the presence of S100 proteins in the stool, serum and bowel tissue were investigated in patients with acute or chronic GvHD and compared to healthy controls and patients without GvHD after HCT. Additionally, the influence of S100 proteins on monocyte-mediated induction of Th17 cells was analysed. The data of this study demonstrate that the expression of S100 proteins is increased in monocytes from patients with GvHD compared to the controls. Overall, elevated levels of S100 proteins can be detected in the serum, stool and bowel tissue of patients with GvHD demonstrating the release of these phagocyte-specific proteins during GvHD. Furthermore, S100 proteins were found to bind to toll-like receptor 4 (TLR4) on monocytes resulting in the promotion of monocyte-induced Th17 development. These data emphasize the role of S100 proteins in Th17 triggered inflammation. Additionally, it was investigated if the induction of Th17 cells is mediated by proinflammatory cytokines released by monocytes or by cell contact between monocytes and CD4+ T cells. The data of the present study have revealed that monocyte-mediated Th17 development occurs in a cell-cell-contact dependent manner with the involvement of proinflammatory cytokines secreted by in vitro or in vivo activated monocytes. A further part of this thesis examined the influence of heat shock protein 90 (Hsp90) in monocyte-mediated induction of Th17 cells. Hsp90 is a ubiquitously expressed molecular chaperon that is known to play an important role in signal transduction, transcription regulation and survival of the cell. The data of this work have demonstrated that Hsp90 inhibition in in vivo activated monocytes by the geldanamycin derivative 17-DMAG decreases Th17 responses. Further results have shown that the stimulatory effects of proinflammatory S100 proteins on monocyte-induced Th17 development can be blocked by chemical inhibition of Hsp90 using 17-DMAG or by specific siRNA-mediated knockdown of the stress-inducible Hsp90α in monocytes. In contrast to dexamethasone which is a potent synthetic member of the glucocorticoid class of steroids that are widely used drugs for the treatment of acute and chronic GvHD, the chemical Hsp90 inhibitor 17-DMAG does not seem to induce the development of proinflammatory Th17 cells expressing the multi-drug resistance protein 1 (MDR1). MDR1 is an ATP-dependent efflux pump that plays a crucial role in the bioavailability of a wide range of drugs and xenobiotics. Altogether, the results of the present work indicate that levels of proinflammatory S100 proteins are increased in the serum, stool and bowel tissue of patients with GvHD and might promote monocyte-induced development of Th17 cells during GvHD. Specific inhibition of Hsp90 might prevent the induction of inflammation-promoting Th17 cells. Therefore, Hsp90 could be a novel, critical target for the treatment of GvHD

Untersuchung der Körperzusammensetzung von AGA-Neugeborenen und SGA-Neugeborenen, sowie Analyse kindlicher und maternaler Einflussparameter Eine prospektive, nicht-interventionelle, klinische Studie unter Verwendung der Air Displacement Plethysmography

Neben dem erhöhten Risiko gesundheitlicher Langzeitfolgen, wie metabolisches Syndrom oder kardiovaskuläre Erkrankungen, besteht bereits in den ersten Lebensstunden eines untergewichtigen bzw. mangelernährten SGA-Kindes eine erhöhte perinatale Mortalität und Morbidität [6]. Von medizinischer Relevanz ist es daher, die mangelernährten Kinder bei Geburt zu identifizieren, um sie gezielt behandeln zu können [156]. Epidemiologische Studien zeigten, dass weder die anthropometrischen Wachstumsparameter noch die Zuordnung zu den Kategorien SGA, AGA und LGA anhand der Wachstumsperzentilen gesicherte Rückschlüsse auf den Ernährungszustand des Kindes erlauben. In dieser prospektiven Studie wurde der Fragestellung nachgegangen, ob sich SGA-Kinder in ihrem BF% von AGA-Kindern unterscheiden. Darüber hinaus wurden mittels Regressionsanalysen mögliche kindliche und maternale Einflussparameter auf den BF% untersucht und sowohl innerhalb als auch zwischen den beiden Gruppen, verglichen (siehe 3.7). Eingeschlossen wurden n=234 eutrophe (AGA) und n=24 hypotrophe Einlings-NG (SGA), die nach der vollendeten 37. SSW geboren wurden und innerhalb der ersten 96 Lebensstunden untersucht wurden. Zur Ermittlung des neonatalen BF% wurde das PEA POD®-System genutzt, das auf dem Verfahren der Air Displacement Plethysmography basiert. Die Auswertung der erhobenen Daten ergab, dass zwischen den AGA- und SGA-Kinder ein statistisch signifikanter Unterschied im BF% bestand. Der mediane BF% der AGA-Gruppe lag bei 11,0% (P25–P75: 8,0%-13,4%), während der mediane BF% der SGA-Gruppe bei 6,6% (P25–P75: 5,2%-8,1%) lag. Trotz statistisch signifikant unterschiedlicher Mediane fiel eine ausgeprägte Überlappung der einzelnen BF%-Werte der beiden Gruppen auf. Nahezu bei 40% der AGA-Kinder lag der BF% unterhalb des höchsten BF%-Werts in der SGA-Gruppe (9,9%). In dieser Arbeit konnte festgestellt werden, dass die auf Basis der SDSGG -Werte durchgeführte Kategorisierung in SGA und AGA die Gefahr birgt, einen erheblichen Anteil an mangelernährten Neonaten bei Geburt nicht zu erfassen, wenn Mangelernährung mittels eines festgelegten Cut-Off-Werts für den BF% definiert wird. In der AGA-Gruppe konnte nachgewiesen werden, dass das Geschlecht einen Einfluss auf den neonatale BF% hat. So hatten weibliche NG mit 11,4% im Vergleich zu männlichen NG mit 9,7% einen signifikant höheren BF%. In der SGA-Gruppe lag der mediane BF% ebenfalls über dem Wert der männlichen NG (6,7% vs. 6,3%), jedoch nicht im statistisch geforderten Signifikanzniveau. Bei der Analyse möglicher maternaler Einflussparameter auf den neonatalen BF% fiel auf, dass bei einer Gewichtszunahme über der Empfehlung der IOM-Richtlinien im Vergleich zu einer empfohlenen Gewichtszunahme, ein höherer medianer BF% der NG vorlag. Dies galt sowohl in der AGA-Gruppe (10,5% vs. 11,4%), als auch in der SGA-Gruppe (5,6% vs. 6,9%). In den bislang verfügbaren Studien konnten für Messungen mit der ADP-Methode noch keine geschlechtsspezifischen BF%-Referenzwerte von AGA-Kindern und gefährdeten Subgruppen, wie SGA-Kindern verschiedener Altersklassen und Ethnien, ermittelt werden. Weitere Studien mit ausreichender Fallzahl und nachfolgende Beobachtungen der untersuchten Kinder bis in das Kindes- und Jugendalter werden benötigt, um relevante BF%-Schwellenwerte für gefährdete Kinder festzulegen und möglichen Folgeerkrankungen präventiv entgegenzuwirken

Primary motor cortex activation and lateralization in patients with tumors of the central region

AbstractHemispheric lateralization is a frequently encountered phenomenon of cortical function. It describes the functional specialization of a region on one side of the brain for a given task. It is well characterized in motor and sensory, as well as language systems and becomes more and more known for various cognitive domains. While in the adult healthy brain hemispheric lateralization is mostly set, pathological processes may lead to cortical reorganization. In these cases neuroplasticity of the corresponding region in the non-dominant hemisphere seems to play an important role. In a previous study we investigated language associated regions in right-handed patients with frontal and temporal tumors of the left hemisphere. We observed a marked change of language lateralization in these patients towards the non-dominant hemisphere as measured by functional MRI (Partovi et al., 2012).In the present study we evaluated activation and lateralization of cortical motor areas in patients with tumors of the central region. BOLD fMRI was performed during unilateral voluntary movements of the contralesional hand in 87 patients. Individual correlations of measured BOLD-signals with the model hemodynamic reference function were determined on a ROI basis in single subjects and compared to those of 16 healthy volunteers. In volunteers the strongest activation is usually found in the M1 hand representation contralateral to the movement, while a weaker homotopic co-activation is observed in ipsilateral M1 (Stippich et al., 2007a). In the patient group our results show significant changes of motor activations, ranging from a reduction of M1 lateralization to equalization of M1 activations or even inversion of M1 lateralization during contralesional movements. This study corroborates in a large patient group the idea that lesions affecting M1 may lead to functional reorganization of cortical motor systems and in particular equalize hemispheric lateralization. However, it is not yet clear whether these changes are only an epiphenomenon or indeed reflect an attempt of recovery of brain function

Masterstudiengang mit integriertem Vorbereitungsdienst in der Metall- und Elektrotechnik - Berufliche Lehrerbildung phasenübergreifend gestalten

Der innovative Masterstudiengang „Berufliche Bildung Integriert“ verbindet die erste und zweite Phase der Lehrerbildung erstmalig konsequent auf Programmebene. Das dreijährige Ausbildungskonzept führt einen Masterstudiengang und einen zweijährigen Vorbereitungsdienst zusammen. Durch die intensive Abstimmung zwischen Universität und Staatlichem Studienseminar sowie die gemeinsam getragene Verantwortung für die Professionalisierung von Lehrkräften ist ein neuer Weg in die Lehrerbildung an beruflichen Schulen implementiert worden. Dieser generiert hohe Synergieeffekte und ist aus qualitativer Sicht zukunftsweisend für die Professionalisierung von Lehrkräften. Das Konzept eröffnet zudem einer bisher nicht hinreichend adressierten Zielgruppe in den Mangelfächern Metalltechnik sowie Elektro- und Informationstechnik den Zugang zum Lehramt an beruflichen Schulen.Schlüsselwörter: Master Integriert, Berufliches Lehramt, Zweites Staatsexamen, Studienseminar, Vorbereitungsdienst_________Innovative Master’s Programme for Engineering Sciences: Vocational Teacher Education linking teaching theory and practice The innovative Master’s programme ‘Integrated Vocational Education’ links teaching theory and practice, which usually take place sequentially, for the very first time within a structured teacher training course. The three-year training concept dovetails a theoretical Master’s degree course with a two year teacher practice course. Intensive coordination between universities and the responsible government institutions (Staatliches Studienseminar), both entities having a joint responsibility for the professional standards to be attained by trainee teachers, has resulted in a new approach being formulated and initiated for the training of teachers at vocational schools. This pioneering approach generates high synergy effects and important future benefits regarding professional standards in vocational teacher training. Moreover, this new and compact Master’s programme addresses the shortage of teachers in two fields, metal technology and electrical engineering & information technology, by directly offering fast track access to a career as a teacher at a vocational school for those people who already hold a BA in Engineering Sciences.Keywords: Master ‘Integrated Vocational Education’, Vocational Teacher Education, teacher practice course, teacher training, Engineering Science

Chromosome-level sequence assembly reveals the structure of the Arabidopsis thaliana Nd-1 genome and its gene set

Pucker B, Holtgräwe D, Stadermann KB, et al. Chromosome-level sequence assembly reveals the structure of the Arabidopsis thaliana Nd-1 genome and its gene set. PLoS One. 2019;14(5): e0216233.In addition to the BAC-based reference sequence of the accession Columbia-0 from the year 2000, several short read assemblies of THE plant model organism Arabidopsis thaliana were published during the last years. Also, a SMRT-based assembly of Landsberg erecta has been generated that identified translocation and inversion polymorphisms between two genotypes of the species. Here we provide a chromosome-arm level assembly of the A. thaliana accession Niederzenz-1 (AthNd-1_v2c) based on SMRT sequencing data. The best assembly comprises 69 nucleome sequences and displays a contig length of up to 16 Mbp. Compared to an earlier Illumina short read-based NGS assembly (AthNd-1_v1), a 75 fold increase in contiguity was observed for AthNd-1_v2c. To assign contig locations independent from the Col-0 gold standard reference sequence, we used genetic anchoring to generate a de novo assembly. In addition, we assembled the chondrome and plastome sequences. Detailed analyses of AthNd-1_v2c allowed reliable identification of large genomic rearrangements between A. thaliana accessions contributing to differences in the gene sets that distinguish the genotypes. One of the differences detected identified a gene that is lacking from the Col-0 gold standard sequence. This de novo assembly extends the known proportion of the A. thaliana pan-genome

Late-Onset Prosthetic Endocarditis with Paraaortic Abscess Caused by Cutibacterium acnes

Cutibacterium acnes, an integral component of the skin’s customary bacterial flora, represents a Gram-positive anaerobic bacterium characterized by its low virulence. Despite its low virulence, the pathogen can cause profound-seated infections as well as infections linked to medical devices. We report a case study of a prosthesis endocarditis accompanied by a paraaortic abscess caused by C. acnes, a development occurring five years prior to composite aortic root and valve replacement. At the point of admission, the patient presented with a combination of symptoms hinting at a subacute progression, such as weight loss, chest pain, and limitations of cardiopulmonary functionality. An anaerobic pathogen, namely C. acnes, was detected in a singular blood culture vial. Since first-line imaging modalities such as echocardiography did not reveal any signs of inflammation, and in the case of a suspected diagnosis for IE, did not show high pretest probability, further diagnostic imaging such as 18F-FDG PET CT was put to use. Here, a highly elevated glucose metabolism around the aortic valve ring was detected, pointing to an inflammatory process. The patient received adjusted intravenous antibiotic therapy over a course of six weeks; he then underwent surgical therapy via re-replacement of the aortic root and valve using a composite conduit. Advanced microbiological analyses, including the amplification of PCR and valve sequencing via 16S rDNA, mainly detected one pathogen: C. acnes. Delayed onset with mild symptoms and laboratory findings is characteristic of infective endocarditis by C. acnes. Due to its high rate of complications, mortality, and morbidity, an infection should not be disregarded as contamination. Recommendations from different studies underline a combination of a positive blood culture and microbiological evidence to differentiate between contamination and true infection in the case of an infection involving C. acnes. Serial blood cultures with prolonged incubation, advanced microbiological analyses, and modified Duke criteria including second-line imaging techniques should be utilized for further evaluation

Despite large choice of effective therapies: Individuals with psoriasis still seem undertreated

Summary Background and objectives Due to the development of new anti‐psoriatic drugs in combination with improved structures for implementation throughout Germany, the medical care of psoriasis patients has markedly improved. In this study we investigated the real‐life utilization of the health care system and identified reasons for dissatisfaction in affected individuals. Patients and methods This non‐interventional cross‐sectional study was conducted as an anonymous online survey from 12/2018 to 01/2019 in Germany. Participants with a self‐reported physician‐confirmed diagnosis of psoriasis and symptoms answered questions about their disease, its influence on daily life and their medical care. Results 649 participants with a mean age of 42.5 ± 13.7 years and equal gender distribution (male: 50.2 %) were evaluated. 54.1 % received medical treatment at the time of the study, 45.9 % did not. Among the participants with medical care, 59.3 % were only moderately or less satisfied with their treatment. Reasons for dissatisfaction with the medication included lack of efficacy and side effects. Participants without medical treatment specified a physician’s lack of time as a main reason for not seeking medical help. Conclusions Despite the availability of efficient therapeutic options in Germany, many individuals with psoriasis are not satisfied. This under‐treated group was identified as a new target population

Time series clustering of T cell subsets dissects heterogeneity in immune reconstitution and clinical outcomes among MUD-HCT patients receiving ATG or PTCy

IntroductionAnti-T-lymphocyte globulin (ATG) or post-transplant cyclophosphamide (PTCy) prevent graft-versus-host disease (GVHD) after hematopoietic cell transplantation (HCT), yet individual patients benefit differentially.MethodsGiven the sparse comparative data on the impact of cellular immune reconstitution in this setting, we studied flow cytometry and clinical outcomes in 339 recipients of 10/10 matched-unrelated donor (MUD) HCT using either ATG (n=304) or PTCy (n=35) for in vivo T cell manipulation along with a haploidentical PTCy control cohort (n=45). Longitudinal cellular immune reconstitution data were analyzed conventionally and with a data science approach using clustering with dynamic time warping to determine the similarity between time-series of T cell subsets.ResultsConsistent with published studies, no significant differences in clinical outcomes were observed at the cohort level between MUD-ATG and MUD-PTCy. However, cellular reconstitution revealed preferences for distinct T cell subpopulations associating with GVHD protection in each setting. Starting early after HCT, MUD-PTCy patients had higher regulatory T cell levels after HCT (p <0.0001), while MUD-ATG patients presented with higher levels of γδ T- or NKT cells (both p <0.0001). Time-series clustering further dissected the patient population’s heterogeneity revealing distinct immune reconstitution clusters. Importantly, it identified phenotypes that reproducibly associated with impaired clinical outcomes within the same in vivo T cell manipulation platform. Exemplarily, patients with lower activated- and αβ T cell counts had significantly higher NRM (p=0.032) and relapse rates (p =0.01).DiscussionThe improved understanding of the heterogeneity of cellular reconstitution in MUD patients with T cell manipulation both at the cohort and individual level may support clinicians in managing HCT complications

Early Exanthema Upon Vemurafenib Plus Cobimetinib Is Associated With a Favorable Treatment Outcome in Metastatic Melanoma: A Retrospective Multicenter DeCOG Study

Background: The combination of BRAF and MEK inhibitors has become standard of care in the treatment of metastatic BRAF V600-mutated melanoma. Clinical factors for an early prediction of tumor response are rare. The present study investigated the association between the development of an early exanthema induced by vemurafenib or vemurafenib plus cobimetinib and therapy outcome. Methods: This multicenter retrospective study included patients with BRAF V600-mutated irresectable AJCC-v8 stage IIIC/D to IV metastatic melanoma who received treatment with vemurafenib (VEM) or vemurafenib plus cobimetinib (COBIVEM). The development of an early exanthema within six weeks after therapy start and its grading according to CTCAEv4.0 criteria was correlated to therapy outcome in terms of best overall response, progression-free (PFS), and overall survival (OS). Results: A total of 422 patients from 16 centers were included (VEM, n=299; COBIVEM, n=123). 20.4% of VEM and 43.1% of COBIVEM patients developed an early exanthema. In the VEM cohort, objective responders (CR/PR) more frequently presented with an early exanthema than non-responders (SD/PD); 59.0% versus 38.7%; p=0.0027. However, median PFS and OS did not differ between VEM patients with or without an early exanthema (PFS, 6.9 versus 6.0 months, p=0.65; OS, 11.0 versus 12.4 months, p=0.69). In the COBIVEM cohort, 66.0% of objective responders had an early exanthema compared to 54.3% of non-responders (p=0.031). Median survival times were significantly longer for patients who developed an early exanthema compared to patients who did not (PFS, 9.7 versus 5.6 months, p=0.013; OS, not reached versus 11.6 months, p=0.0061). COBIVEM patients with a mild early exanthema (CTCAEv4.0 grade 1-2) had a superior survival outcome as compared to COBIVEM patients with a severe (CTCAEv4.0 grade 3-4) or non early exanthema, respectively (p=0.047). This might be caused by the fact that 23.6% of patients with severe exanthema underwent a dose reduction or discontinuation of COBIVEM compared to only 8.9% of patients with mild exanthema. Conclusions: The development of an early exanthema within 6 weeks after treatment start indicates a favorable therapy outcome upon vemurafenib plus cobimetinib. Patients presenting with an early exanthema should therefore be treated with adequate supportive measures to provide that patients can stay on treatment