Vectorial import via a metastable disulfide-linked complex allows for a quality control step and import by the mitochondrial disulfide relay

Disulfide formation in the mitochondrial intermembrane space (IMS) is an essential process. It is catalyzed by the disulfide relay machinery, which couples substrate import and oxidation. The machinery relies on the oxidoreductase and chaperone CHCHD4-Mia40. Here, we report on the driving force for IMS import and on a redox quality control mechanism. We demonstrate that unfolded reduced proteins, upon translocation into the IMS, initiate formation of a metastable disulfide-linked complex with CHCHD4. If this interaction does not result in productive oxidation, then substrates are released to the cytosol and degraded by the proteasome. Based on these data, we propose a redox quality control step at the level of the disulfide-linked intermediate that relies on the vectorial nature of IMS import. Our findings also provide the mechanistic framework to explain failures in import of numerous human disease mutants in CHCHD4 substrates

Akute Transiente atmungsbedingte Bewegungsartefakte nach Gadoxetat-Applikation in der arteriellen Magnetresonanztomographie der Leber

In dieser Dissertation wurden Transient Severe Motion (TSM) Artefakte in der mit Gadoxetat kontrastierten Magnetresonanztomographie (MRT) der Leber untersucht. Dieses Phänomen beeinträchtigt die Diagnostik erheblich. Erforscht wurde die Häufigkeit der TSM-Artefakte, der Einfluss des genauen Zeitabschnitts der bildlichen Erfassung der arteriellen Kontrastierungsphase auf ihr Auftreten sowie Risikofaktoren dafür. 354 Patienten, die von 2013 bis 2016 am Institut für Diagnostische Radiologie und Neuroradiologie der Universität Greifswald untersucht wurden, gingen in die retrospektive Studie ein. 69 dieser Patienten erhielten eine Folgeuntersuchung. Aufnahmen der arteriellen Phase wurden hinsichtlich der TSM-Artefakte auf einer vierstufigen Skala nach Schweregrad eingeteilt (0=keine TSM, 1=leichte TSM, 2=mäßige TSM, 3=schwere TSM). Das Auftreten von TSM-Artefakten war erheblich vom genauen Zeitabschnitt der bildlichen Erfassung der arteriellen Phase (früharteriell, vollarteriell, spätarteriell) sowie eventueller TSM-Artefakte in Voruntersuchungen und verschiedenen Risikofaktoren abhängig. TSM-Artefakte traten bei 48,59% der untersuchten Patienten auf (172/354). Bei 22,88% aller Patienten (n=81) traten TSM-Artefakte des Grades 1 auf. Bei 18,36% (n=65) wurden TSM-Artefakte des Grades 2 festgestellt. Bei 7,34% (n=26) kam es zu TSM-Artefakten des Grades 3. In der vollarteriellen Phase kam es mit 58,8% der Untersuchungen am häufigsten zu TSM-Artefakten. In der früharteriellen Phase (51,6%) sowie der spätarteriellen Phase (42,1%) traten die Artefakte signifikant seltener auf (p=0,031). Das Auftreten von TSM-Artefakten korrelierte signifikant mit dem BMI (p=0,001) sowie grenzwertig mit Leberzirrhose (p=0,05). TSM-Artefakte der verschiedenen Schweregrade korrelierten signifikant mit Gewicht (p=0,03), Größe (p=0,033), BMI (p=0,003) sowie γ-GT (p=0,029). Bei Folgeuntersuchungen zeigte sich ein signifikanter Zusammenhang zwischen TSM-Artefakten in der Erstuntersuchung und TSM-Artefakten in Folgeuntersuchungen (p=0,041). In dieser Studie konnte gezeigt werden, dass die geringste Wahrscheinlichkeit für TSM-Artefakte in der spätarteriellen Phase besteht. Daher ist bei der Diagnostik der mit Gadoxetat kontrastierten Leber-MRT die spätarterielle Phase zu empfehlen

p62 Promotes Survival and Hepatocarcinogenesis in Mice with Liver-Specific NEMO Ablation

SQSTM1/p62 is a multitasking protein that functions as an autophagy receptor, but also as a signaling hub regulating diverse cellular pathways. p62 accumulation in mice with autophagydeficient hepatocytes mediates liver damage and hepatocarcinogenesis through Nrf2 overactivation, yet the role of the p62-Keapl-Nrf2 axis in cell death and hepatocarcinogenesis in the absence of underlying autophagy defects is less clear. Here, we addressed the role of p62 and Nrf2 activation in a chronic liver disease model, namely mice with liver parenchymal cell-specific knockout of NEMO (NEMOLPC-KO), in which we demonstrate that they show no inherent autophagy impairment. Unexpectedly, systemic p62 ablation aggravated the phenotype and caused early postnatal lethality in NEMO(LPC-KO )mice. Expression of a p62 mutant (p624Ex2-5), which retains the ability to form aggregates and activate Nrf2 signaling, did not cause early lethality, but exacerbated hepatocarcinogenesis in these mice. Our immunohistological and molecular analyses showed that the increased tumor burden was only consistent with increased expression/stability of p624Ex2-5 driving Nrf2 hyperactivation, but not with other protumorigenic functions of p62, such as mTOR activation, cMYC upregulation or increased fibrosis. Surprisingly, forced activation of Nrf2 per se did not increase liver injury or tumor burden in NEMOLPC-KO mice, suggesting that autophagy impairment is a necessary prerequisite to unleash the Nrf2 oncogenic potential in mice with autophagy-competent hepatocytes.ISSN:2072-669

XIAP promotes melanoma growth by inducing tumour neutrophil infiltration

Elevated expression of the X-linked inhibitor of apoptosis protein (XIAP) has been frequently reported in malignant melanoma suggesting that XIAP renders apoptosis resistance and thereby supports melanoma progression. Independent of its anti-apoptotic function, XIAP mediates cellular inflammatory signalling and promotes immunity against bacterial infection. The pro-inflammatory function of XIAP has not yet been considered in cancer. By providing detailed in vitro analyses, utilising two independent mouse melanoma models and including human melanoma samples, we show here that XIAP is an important mediator of melanoma neutrophil infiltration. Neutrophils represent a major driver of melanoma progression and are increasingly considered as a valuable therapeutic target in solid cancer. Our data reveal that XIAP ubiquitylates RIPK2, involve TAB1/RIPK2 complex and induce the transcriptional up-regulation and secretion of chemokines such as IL8, that are responsible for intra-tumour neutrophil accumulation. Alteration of the XIAP-RIPK2-TAB1 inflammatory axis or the depletion of neutrophils in mice reduced melanoma growth. Our data shed new light on how XIAP contributes to tumour growth and provides important insights for novel XIAP targeting strategies in cancer

Elevated X-linked inhibitor of apoptosis protein (XIAP) expression uncovers detrimental prognosis in subgroups of neoadjuvant treated and T-cell rich esophageal adenocarcinoma

Background: Molecular markers predicting survival in esophageal adenocarcinoma (EAC) are rare. Specifically, in favorable oncologic situations, e.g. nodal negativity or major neoadjuvant therapy response, there is a lack of additional risk factors that serve to predict patients' outcome more precisely. This study evaluated X-linked inhibitor of apoptosis protein (XIAP) as a potential marker improving outcome prediction. Methods: Tissue microarrays from 362 patients that were diagnosed with resectable EAC were included in the study. XIAP was stained by immunohistochemistry and correlated to clinical outcome, molecular markers and markers of the cellular tumor microenvironment. Results: XIAP did not impact on overall survival (OS) in the whole study collective. Subgroup analyses stratifying for common genetic markers (TP53, ERBB2, ARID1A/SWI/SNF) did not disclose any impact of XIAP expression on survival. Detailed subgroup analyses of [1] nodal negative patients, [2] highly T-cell infiltrated tumors and [3] therapy responders to neoadjuvant treatment revealed a significant inverse role of high XIAP expression in these specific oncologic situations; elevated XIAP expression detrimentally affected patients' outcome in these subgroups. [1]: OS XIAP low: 202 months (m) vs. XIAP high: 38 m; [2]: OS 116 m vs. 28.2m; [3]: OS 31 m vs. 4 m). Conclusions: Our data suggest XIAP expression in EAC as a worthy tool to improve outcome prediction in specific oncologic settings that might directly impact on clinical diagnosis and treatment of EAC in the future

Application of fluorine nmr for structure identification of steroids

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