Selective Induction of Apoptosis in Melanoma Cells by Tyrosinase Promoter-Controlled CD95 Ligand Overexpression

Induction of apoptosis has been demonstrated previously by overexpression of CD95 ligand (CD95L) in cultured human melanoma cells. For in vivo approaches based on CD95L, however, targeted expression is a prerequisite and tyrosinase promoters have been considered for selection. Luciferase reporter gene assays performed for a representative panel of melanoma cell lines characterized by strong (SK-Mel-19), moderate (SK-Mel-13, MeWo), weak (A-375), and missing expression (M-5) of endogenous tyrosinase revealed high tyrosinase promoter activities in SK-Mel-19, SK-Mel-13, and MeWo, but only weak activities in A-375 and M-5 as well as in non-melanoma cell lines. After transfection of a CMV promoter CD95L expression construct, melanoma cells were found highly sensitive, as compared with non-melanoma cells. By applying a tyrosinase promoter CD95L construct, apoptosis was selectively induced in SK-Mel-19, SK-Mel-13, MeWo as well as in A-375, which was characterized by high CD95 surface expression and high sensitivity to agonistic CD95 activation. M5 and non-melanoma cell lines remained uninfluenced. Also, resistance to agonistic CD95 activation seen in MeWo characterized by weak CD95 surface expression was overcome by overexpression of CD95L. Our investigations provide evidence that tyrosinase promoter CD95L constructs may be of value for selective induction of apoptosis in therapeutic strategies for melanoma

Anreizsysteme - Eine Möglichkeit zur Verbesserung der universitären Lehre?

Eingeleitet wird das Journal mit dem Beitrag "Anreize für gute Lehre" von Kiefer, Niederhaus, Balzani, Bobisch, Gerharz, Kruggel-Emden, Schwarz, Thielbörger & Weiss, Mitgliedern der Global Young Faculty. In ihrer Umfrage in den UMAR-Universitäten gingen sie der Frage nach "Was motiviert Lehrende qualitativ gute Lehre anzubieten?". Ihre Ergebnisse überraschen und zeigen, welchen Stellenwert nach Meinung der Interviewten Lehre generell einnimmt

Reduced Calcium Signaling Is Associated With Severe Graft-Versus-Host Disease: Results From Preclinical Models and From a Prospective EBMT Study

Despite its involvement in various immune functions, including the allogeneic activation of T-lymphocytes, the relevance of calcium (Ca2+) for GVHD pathobiology is largely unknown. To elucidate a potential association between Ca(2+)and GVHD, we analyzed Ca2+-sensing G-protein coupled receptor 6a (GPRC6a) signaling in preclinical GVHD models and conducted a prospective EBMT study on Ca(2+)serum levels prior alloSCT including 363 matched sibling allogeneic peripheral blood stem cell transplantations (alloSCTs). In experimental models, we found decreasedGprc6aexpression during intestinal GVHD. GPRC6a deficient alloSCT recipients had higher clinical and histopathological GVHD scores leading to increased mortality. As possible underlying mechanism, we found increased antigen presentation potential in GPRC6a(-/-)alloSCT recipients demonstrated by higher proliferation rates of T-lymphocytes. In patients with low Ca(2+)serum levels (≤ median 2.2 mmol/l) before alloSCT, we found a higher incidence of acute GVHD grades II-IV (HR = 2.3 Cl = 1.45-3.85p= 0.0006), severe acute GVHD grades III-IV (HR = 3.3 CI = 1.59-7.14,p= 0.002) and extensive chronic GVHD (HR = 2.0 Cl = 1.04-3.85p= 0.04). In conclusion, experimental and clinical data suggest an association of reduced Ca(2+)signaling with increased severity of GVHD. Future areas of interest include the in depth analysis of involved molecular pathways and the investigation of Ca(2+)signaling as a therapeutic target during GVHD

Characterisation and use of β-lactoglobulin fibrils for microencapsulation of lipophilic ingredients and oxidative stability thereof

There is a growing interest in using fibrils from food grade protein, e.g. β-lactoglobulin, as functional ingredients. In the present study, the functionality of fibrillar β-lactoglobulin from whey protein isolate (WPI) was compared to native WPI in terms of interfacial dilatational rheology and emulsifying activity at acidic conditions (pH 2.0 and 3.0). We report here for the first time data on microencapsulation of fish oil by spray-drying as well as oxidative stability of the oil in emulsions and microcapsules in dependence of WPI conformation. WPI fibrils exerted a significantly higher elasticity at the oil–water (o/w) interface and a better emulsifying activity at a fixed oil content compared to native WPI. Microencapsulation efficiency was also higher with fibrillar WPI (>95%) compared to native WPI (∼90%) at pH 2.0 and a total oil and protein content of 40% and 2.2%, respectively, in the final powder. The oxidative deterioration was lower in emulsions and microcapsules prepared with fibrillar than with native WPI. This was attributed to improved interfacial barrier properties provided by fibrils and antioxidative effects of coexisting unconverted monomers, particularly hydrophilic peptides

Premature termination, satisfaction with care, and shared decision-making during home treatment compared to inpatient treatment: A quasi-experimental trial

Abstract Background Inpatient equivalent home treatment (IEHT), implemented in Germany since 2018, is a specific form of home treatment. Between 2021 and 2022, IEHT was compared to inpatient psychiatric treatment in a 12-months follow-up quasi-experimental study with two propensity score matched cohorts in 10 psychiatric centers in Germany. This article reports results on the treatment during the acute episode and focuses on involvement in decision-making, patient satisfaction, and drop-out rates. Methods A total of 200 service users receiving IEHT were compared with 200 matched statistical “twins” in standard inpatient treatment. Premature termination of treatment as well as reasons for this was assessed using routine data and a questionnaire. In addition, we measured patient satisfaction with care with a specific scale. For the evaluation of patient involvement in treatment decisions, we used the 9-item Shared Decision Making Questionnaire (SDM-Q-9). Results Patients were comparable in both groups with regard to sociodemographic and clinical characteristics. Mean length-of-stay was 37 days for IEHT and 28 days for inpatient treatment. In both groups, a similar proportion of participants stopped treatment prematurely. At the end of the acute episode, patient involvement in decision-making (SDM-Q-9) as well as treatment satisfaction scores were significantly higher for IEHT patients compared to inpatients. Conclusions Compared to inpatient care, IEHT treatment for acute psychiatric episodes was associated with higher treatment satisfaction and more involvement in clinical decisions

Erfahrungen, Herausforderungen und Lösungsansätze aus der Extraktion pseudonymer Daten für das Projekt INDEED

Background: In Germany there is currently no health reporting on cross-sectoral care patterns in the context of an emergency department care treatment. The INDEED project (Utilization and trans-sectoral patterns of care for patients admitted to emergency departments in Germany) collects routine data from 16 emergency departments, which are later merged with outpatient billing data from 2014 to 2017 on an individual level. Aim: The methodological challenges in planning of the internal merging of routine clinical and administrative data from emergency departments in Germany up to the final data extraction are presented together with possible solution approaches. Methods: Data were selected in an iterative process according to the research questions, medical relevance, and assumed data availability. After a preparatory phase to clarify formalities (including data protection, ethics), review test data and correct if necessary, the encrypted and pseudonymous data extraction was performed. Results: Data from the 16 cooperating emergency departments came mostly from the emergency department and hospital information systems. There was considerable heterogeneity in the data. Not all variables were available in every emergency department because, for example, they were not standardized and digitally available or the extraction effort was judged to be too high. Conclusion: Relevant data from emergency departments are stored in different structures and in several IT systems. Thus, the creation of a harmonized data set requires considerable resources on the part of the hospital as well as the data processing unit. This needs to be generously calculated for future projects

Metabolic Deficiences Revealed in the Biotechnologically Important Model Bacterium Escherichia coli BL21(DE3)

The Escherichia coli B strain BL21(DE3) has had a profound impact on biotechnology through its use in the production of recombinant proteins. Little is understood, however, regarding the physiology of this important E. coli strain. We show here that BL21(DE3) totally lacks activity of the four [NiFe]-hydrogenases, the three molybdenum- and selenium-containing formate dehydrogenases and molybdenum-dependent nitrate reductase. Nevertheless, all of the structural genes necessary for the synthesis of the respective anaerobic metalloenzymes are present in the genome. However, the genes encoding the high-affinity molybdate transport system and the molybdenum-responsive transcriptional regulator ModE are absent from the genome. Moreover, BL21(DE3) has a nonsense mutation in the gene encoding the global oxygen-responsive transcriptional regulator FNR. The activities of the two hydrogen-oxidizing hydrogenases, therefore, could be restored to BL21(DE3) by supplementing the growth medium with high concentrations of Ni2+ (Ni2+-transport is FNR-dependent) or by introducing a wild-type copy of the fnr gene. Only combined addition of plasmid-encoded fnr and high concentrations of MoO42− ions could restore hydrogen production to BL21(DE3); however, to only 25–30% of a K-12 wildtype. We could show that limited hydrogen production from the enzyme complex responsible for formate-dependent hydrogen evolution was due solely to reduced activity of the formate dehydrogenase (FDH-H), not the hydrogenase component. The activity of the FNR-dependent formate dehydrogenase, FDH-N, could not be restored, even when the fnr gene and MoO42− were supplied; however, nitrate reductase activity could be recovered by combined addition of MoO42− and the fnr gene. This suggested that a further component specific for biosynthesis or activity of formate dehydrogenases H and N was missing. Re-introduction of the gene encoding ModE could only partially restore the activities of both enzymes. Taken together these results demonstrate that BL21(DE3) has major defects in anaerobic metabolism, metal ion transport and metalloprotein biosynthesis

SAMHD1 is a biomarker for cytarabine response and a therapeutic target in acute myeloid leukemia.

The nucleoside analog cytarabine (Ara-C) is an essential component of primary and salvage chemotherapy regimens for acute myeloid leukemia (AML). After cellular uptake, Ara-C is converted into its therapeutically active triphosphate metabolite, Ara-CTP, which exerts antileukemic effects, primarily by inhibiting DNA synthesis in proliferating cells. Currently, a substantial fraction of patients with AML fail to respond effectively to Ara-C therapy, and reliable biomarkers for predicting the therapeutic response to Ara-C are lacking. SAMHD1 is a deoxynucleoside triphosphate (dNTP) triphosphohydrolase that cleaves physiological dNTPs into deoxyribonucleosides and inorganic triphosphate. Although it has been postulated that SAMHD1 sensitizes cancer cells to nucleoside-analog derivatives through the depletion of competing dNTPs, we show here that SAMHD1 reduces Ara-C cytotoxicity in AML cells. Mechanistically, dGTP-activated SAMHD1 hydrolyzes Ara-CTP, which results in a drastic reduction of Ara-CTP in leukemic cells. Loss of SAMHD1 activity-through genetic depletion, mutational inactivation of its triphosphohydrolase activity or proteasomal degradation using specialized, virus-like particles-potentiates the cytotoxicity of Ara-C in AML cells. In mouse models of retroviral AML transplantation, as well as in retrospective analyses of adult patients with AML, the response to Ara-C-containing therapy was inversely correlated with SAMHD1 expression. These results identify SAMHD1 as a potential biomarker for the stratification of patients with AML who might best respond to Ara-C-based therapy and as a target for treating Ara-C-refractory AML

Das Gefäßsystem und die Immunzellrekonstitution in der Knochenmarksnische nach allogener hämatopoetischer Stammzelltransplantation

For a variety of malignant diseases of the hematopoietic system, allogeneic hematopoietic stem cell transplantation (allo-HSCT) is the only curative treatment option. Unfortunately, more than half of the patients receiving allo-HSCT die within the first two years. A major reason for mortality after allo-HSCT is acute graft-versus-host disease (aGVHD), a systemic inflammatory disease in which activated donor T cells damage the host tissue. Patients suffering from aGVHD often develop severe prolonged cytopenia with serious complications such as heavy bleedings, fatal infections, and tumor relapse. This association suggests that the bone marrow (BM) niche, which is critical for hematopoiesis, plays a decisive role in the outcome after allo-HSCT. However, little is known about the role of the BM niche as an aGVHD target tissue, and the underlying pathophysiology of aGVHD-mediated BM niche damage is unclear. This study aimed to unravel the influence of aGVHD on the BM vasculature and BM immune cell reconstitution as selected BM niche features. For this purpose, a clinically relevant allo-HSCT model for BM aGVHD based on the transplantation of purified hematopoietic stem cells (Lineage−Sca-1+c-Kit+ cells) was established. Experimental allo-HSCT led to profound engraftment and induced typical features of aGVHD. The descriptive overview of the above-mentioned BM niche characteristics revealed an increased BM vessel density at day+20 after allo-HSCT and a typical delayed immune cell reconstitution in the BM of allo-HSCT recipients during aGVHD. Further, the effect of CD31+ cells as a therapeutic approach to aGVHD clinics, BM vessel density, and BM immune cell reconstitution after allo-HSCT was tested. The intravenous CD31+ cell transfer resulted in improved clinical appearance shown by lower aGVHD scores at day+23 after allo-HSCT, and alterations of the BM immune cells towards reduced CD4+ T cells as well as CD4+ effector memory T cells. The findings obtained underline that aGVHD affects the BM and add preliminary data of CD31+ cell transfer as a possible therapeutic option for aGVHD. This study provides the foundation from which further development, such as mechanistic analyses as well as translational development of therapeutic approaches, can progress.Für eine Vielzahl von bösartigen Erkrankungen des hämatopoetischen Systems stellt die allogene hämatopoetische Stammzelltransplantation (allo-HSZT) die einzige kurative Behandlungsmöglichkeit dar. Leider sterben mehr als die Hälfte der Empfänger einer allo-HSZT innerhalb der ersten zwei Jahre. Ein Hauptgrund für die Mortalität nach allo-HSZT ist die akute graft-versus-host Krankheit (aGVHD). Die aGVHD ist eine systemische Entzündungskrankheit, bei der aktivierte T-Zellen des Spenders das Wirtsgewebe schädigen. Patienten, die an aGVHD leiden, entwickeln häufig eine schwere, langanhaltende Zytopenie mit erheblichen Komplikationen wie starken Blutungen, fatalen Infekten und Tumorrezidiven. Diese Assoziation deutet darauf hin, dass die Knochenmarksnische, welche von wesentlicher Bedeutung für die Hämatopoese ist, eine entscheidende Rolle für das Ergebnis nach allo-HSCT spielt. Über die Bedeutung und Wirkung der Knochenmarksnische als aGVHD-Zielgewebe ist jedoch wenig bekannt, und die zu Grunde liegende Pathophysiologie der aGVHD-vermittelten Knochenmarksnischenschädigung ist unklar. Ziel dieser Studie war es, den Einfluss der aGVHD auf die Gefäßdichte und die Immunzellrekonstitution im Knochenmark (KM) als ausgewählte KM-Nischenmerkmale zu untersuchen. Hierfür wurde zunächst ein klinisch relevantes allo-HSZT-Modell für die KM aGVHD etabliert, welches sich durch die Transplantation von aufbereiteten hämatopoetischen Stammzellen (Lineage−Sca-1+c-Kit+ Zellen) auszeichnet. Die experimentelle allo-HSZT führte zu einem sicheren Engraftment und induzierte typische Merkmale der aGVHD. Die Aufstellung eines beschreibenden Überblicks über die genannten KM-Nischenmerkmale ergab eine erhöhte KM-Gefäßdichte an Tag+20 nach allo-HSZT und eine typisch verzögerte Immunzellrekonstitution im KM von allo-HSZT Empfängern während der aGVHD. Darüber hinaus wurde der Einfluss von CD31+ Zellen als therapeutischer Ansatz auf die aGVHD Klinik, die KM-Gefäßdichte und die KM-Immunzellrekonstitution nach allo-HSZT getestet. Der intravenöse Transfer von CD31+ Zellen führte zu einem verbesserten klinischen Erscheinungsbild, das sich durch niedrigere aGVHD Scores an Tag+23 nach allo-HSZT darstellte, zu reduzierten CD4+ T Zellen, sowie reduzierten CD4+ T Effektor-Gedächtnis Zellen. Die gewonnenen Erkenntnisse unterstreichen, dass die aGVHD das KM beeinflusst und zeigen erste Daten von einem CD31+ Zell-Transfer als möglicher Therapieoption. Diese Studie liefert die Grundlage, auf der weitere Entwicklungen, wie mechanistische Analysen sowie die Entwicklung von therapeutischen Ansätzen, entstehen können

Don’t Let it Get to You! A Moderated Mediated Approach to the (In)justice–Health Relationship

The present study investigates the consequences of overall justice perceptions on employees’ mental health and work–family conflict. While many studies have found that perceiving injustice at work is harmful, little is known about the underlying processes. Based on the allostatic load model, it is hypothesized that mental preoccupation with work, defined as a cognitive state, is a mediator linking overall justice perceptions to employee health. Moreover, we argue that locus of control is a moderator for the mediated relationship. We tested our hypotheses with panel data consisting of 412 Swedish office workers. Results support that mental preoccupation with work mediates the relationship between overall justice and mental health, and overall justice and work–family conflict. Results also reveal that mental preoccupation with work plays a greater mediating role for individuals with an external locus of control. Implications and suggestions for future studies on the emerging relationship between organizational justice and health are discussed