Monitoring resilience in a rook-managed containerized cloud storage system

Distributed cloud storage solutions are currently gaining high momentum in industry and academia. The enterprise data volume growth and the recent tendency to move as much as possible data to the cloud is strongly stimulating the storage market growth. In this context, and as a main requirement for cloud native applications, it is of utmost importance to guarantee resilience of the deployed applications and the infrastructure. Indeed, with failures frequently occurring, a storage system should quickly recover to guarantee service availability. In this paper, we focus on containerized cloud storage, proposing a resilience monitoring solution for the recently developed Rook storage operator. While, Rook brings storage systems into a cloud-native container platform, in this paper we design an additional module to monitor and evaluate the resilience of the Rook-based system. Our proposed module is validated in a production environment, with software components generating a constant load and a controlled removal of system elements to evaluate the self-healing capability of the storage system. Failure recovery time revealed to be 41 and 142 seconds on average for a 32GB and a 215GB object storage device respectively

SOAT1: a suitable target for therapy in high-grade astrocytic glioma?

Targeting molecular alterations as an effective treatment for isocitrate dehydrogenase-wildtype glioblastoma (GBM) patients has not yet been established. Sterol-O-Acyl Transferase 1 (SOAT1), a key enzyme in the conversion of endoplasmic reticulum cholesterol to esters for storage in lipid droplets (LD), serves as a target for the orphan drug mitotane to treat adrenocortical carcinoma. Inhibition of SOAT1 also suppresses GBM growth. Here, we refined SOAT1-expression in GBM and IDH-mutant astrocytoma, CNS WHO grade 4 (HGA), and assessed the distribution of LD in these tumors. Twenty-seven GBM and three HGA specimens were evaluated by multiple GFAP, Iba1, IDH1 R132H, and SOAT1 immunofluorescence labeling as well as Oil Red O staining. To a small extent SOAT1 was expressed by tumor cells in both tumor entities. In contrast, strong expression was observed in glioma-associated macrophages. Triple immunofluorescence labeling revealed, for the first time, evidence for SOAT1 colocalization with Iba1 and IDH1 R132H, respectively. Furthermore, a notable difference in the amount of LD between GBM and HGA was observed. Therefore, SOAT1 suppression might be a therapeutic option to target GBM and HGA growth and invasiveness. In addition, the high expression in cells related to neuroinflammation could be beneficial for a concomitant suppression of protumoral microglia/macrophages

SOAT1: A Suitable Target for Therapy in High-Grade Astrocytic Glioma?

Targeting molecular alterations as an effective treatment for isocitrate dehydrogenasewildtype glioblastoma (GBM) patients has not yet been established. Sterol-O-Acyl Transferase 1 (SOAT1), a key enzyme in the conversion of endoplasmic reticulum cholesterol to esters for storage in lipid droplets (LD), serves as a target for the orphan drug mitotane to treat adrenocortical carcinoma. Inhibition of SOAT1 also suppresses GBM growth. Here, we refined SOAT1-expression in GBM and IDH-mutant astrocytoma, CNS WHO grade 4 (HGA), and assessed the distribution of LD in these tumors. Twenty-seven GBM and three HGA specimens were evaluated by multiple GFAP, Iba1, IDH1 R132H, and SOAT1 immunofluorescence labeling as well as Oil Red O staining. To a small extent SOAT1 was expressed by tumor cells in both tumor entities. In contrast, strong expression was observed in glioma-associated macrophages. Triple immunofluorescence labeling revealed, for the first time, evidence for SOAT1 colocalization with Iba1 and IDH1 R132H, respectively. Furthermore, a notable difference in the amount of LD between GBM and HGA was observed. Therefore, SOAT1 suppression might be a therapeutic option to target GBM and HGA growth and invasiveness. In addition, the high expression in cells related to neuroinflammation could be beneficial for a concomitant suppression of protumoral microglia/macrophages

Heatwave-associated Vibrio infections in Germany, 2018 and 2019

Background: Vibrio spp. are aquatic bacteria that prefer warm seawater with moderate salinity. In humans, they can cause gastroenteritis, wound infections, and ear infections. During the summers of 2018 and 2019, unprecedented high sea surface temperatures were recorded in the German Baltic Sea. Aim: We aimed to describe the clinical course and microbiological characteristics of Vibrio infections in Germany in 2018 and 2019. Methods: We performed an observational retrospective multi-centre cohort study of patients diagnosed with domestically-acquired Vibrio infections in Germany in 2018 and 2019. Demographic, clinical, and microbiological data were assessed, and isolates were subjected to whole genome sequencing and antimicrobial susceptibility testing. Results: Of the 63 patients with Vibrio infections, most contracted the virus between June and September, primarily in the Baltic Sea: 44 (70%) were male and the median age was 65 years (range: 2–93 years). Thirty-eight patients presented with wound infections, 16 with ear infections, six with gastroenteritis, two with pneumonia (after seawater aspiration) and one with primary septicaemia. The majority of infections were attributed to V. cholerae (non–O1/non-O139) (n = 30; 48%) or V. vulnificus (n = 22; 38%). Phylogenetic analyses of 12 available isolates showed clusters of three identical strains of V. vulnificus, which caused wound infections, suggesting that some clonal lines can spread across the Baltic Sea. Conclusions: During the summers of 2018 and 2019, severe heatwaves facilitated increased numbers of Vibrio infections in Germany. Since climate change is likely to favour the proliferation of these bacteria, a further increase in Vibrio-associated diseases is expected.Peer Reviewe

Klebsiella pneumoniae exhibiting a phenotypic hyper-splitting phenomenon including the formation of small colony variants

In this study, we characterized a Klebsiella pneumoniae strain in a patient with shrapnel hip injury, which resulted in multiple phenotypic changes, including the formation of a small colony variant (SCV) phenotype. Although already described since the 1960s, there is little knowledge about SCV phenotypes in Enterobacteriaceae. The formation of SCVs has been recognized as a bacterial strategy to evade host immune responses and compromise the efficacy of antimicrobial therapies, leading to persistent and recurrent courses of infections. In this case, 14 isolates with different resisto- and morpho-types were distinguished from the patient’s urine and tissue samples. Whole genome sequencing revealed that all isolates were clonally identical belonging to the K. pneumoniae high-risk sequence type 147. Subculturing the SCV colonies consistently resulted in the reappearance of the initial SCV phenotype and three stable normal-sized phenotypes with distinct morphological characteristics. Additionally, an increase in resistance was observed over time in isolates that shared the same colony appearance. Our findings highlight the complexity of bacterial behavior by revealing a case of phenotypic “hyper-splitting” in a K. pneumoniae SCV and its potential clinical significance

Framework and baseline examination of the German National Cohort (NAKO)

The German National Cohort (NAKO) is a multidisciplinary, population-based prospective cohort study that aims to investigate the causes of widespread diseases, identify risk factors and improve early detection and prevention of disease. Specifically, NAKO is designed to identify novel and better characterize established risk and protection factors for the development of cardiovascular diseases, cancer, diabetes, neurodegenerative and psychiatric diseases, musculoskeletal diseases, respiratory and infectious diseases in a random sample of the general population. Between 2014 and 2019, a total of 205,415 men and women aged 19–74 years were recruited and examined in 18 study centres in Germany. The baseline assessment included a face-to-face interview, self-administered questionnaires and a wide range of biomedical examinations. Biomaterials were collected from all participants including serum, EDTA plasma, buffy coats, RNA and erythrocytes, urine, saliva, nasal swabs and stool. In 56,971 participants, an intensified examination programme was implemented. Whole-body 3T magnetic resonance imaging was performed in 30,861 participants on dedicated scanners. NAKO collects follow-up information on incident diseases through a combination of active follow-up using self-report via written questionnaires at 2–3 year intervals and passive follow-up via record linkages. All study participants are invited for re-examinations at the study centres in 4–5 year intervals. Thereby, longitudinal information on changes in risk factor profiles and in vascular, cardiac, metabolic, neurocognitive, pulmonary and sensory function is collected. NAKO is a major resource for population-based epidemiology to identify new and tailored strategies for early detection, prediction, prevention and treatment of major diseases for the next 30 years. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10654-022-00890-5

Uniform nomenclature for the mitochondrial contact site and cristae organizing system

The mitochondrial inner membrane contains a large protein complex that functions in inner membrane organization and formation of membrane contact sites. The complex was variably named the mitochondrial contact site complex, mitochondrial inner membrane organizing system, mitochondrial organizing structure, or Mitofilin/Fcj1 complex. To facilitate future studies, we propose to unify the nomenclature and term the complex "mitochondrial contact site and cristae organizing system" and its subunits Mic10 to Mic60

Molekulare Mechanismen der Osmoregulation in Magnaporthe oryzae und deren Modulation durch das Fungizid Fludioxonil

Zusammenfassung Die stetig größer werdende Herausforderung und unausweichliche, globale Problematik zunehmender Fungizidresistenzen phytopathogener Pilze ist hochaktuell. Das Fungizid Fludioxonil, welches den für die pilzliche Osmoregulation verantwortlichen „High Osmolarity Glycerol“ (HOG)-Signalweg moduliert, wird seit mehr als 20 Jahren weltweit in der Landwirtschaft eingesetzt – bisher ohne nennenswerte Resistenzbildungen. Der molekulare Wirkmechanismus dieses Fungizids ist noch weitgehend unverstanden und dessen Erforschung deshalb von höchster Relevanz für den integrierten Pflanzenschutz. Die im Rahmen dieser Arbeit mit dem filamentösen Ascomycet Magnaporthe oryzae (M. oryzae) erzielten Ergebnisse zu molekularen Mechanismen der Wirkungsweise von Fludioxonil und zu sensorischen Funktionen von Zweikomponenten-Hybrid-Histidinkinasen (HKs) werden dazu einen wertvollen Beitrag liefern. Es konnte eindeutig nachgewiesen werden, dass es neben Genen, welche bisher bekannte Proteine des HOG-Signalwegs von M. oryzae kodieren, noch weitere Komponenten gibt, deren genetische Manipulation zu einer veränderten Fludioxonil-Suszeptibilität in filamentösen, phytopathogenen Pilzen führt. Die homologe Überexpression der Phosphatase MoPtp2p resultierte in einer Fludioxonil-resistenten Mutante mit reduziertem Phosphorylierungsgrad der Mitogen-aktivierte Proteinkinase (MAPK) MoHog1p. Inaktivierung von MoPTP2 resultierte hingegen in einem Fludioxonil-sensitiven Phänotyp. Daraus kann geschlossen werden, dass MoPtp2p am Prozess der Signaltransduktion im HOG-Signalweg und auch entscheidend an der Wirkungsweise des Fungizids Fludioxonil beteiligt ist. Die für die Osmoregulation essentielle MAPK MoHog1p transloziert nach Aktivierung vom Zytosol in den Zellkern. Durch Proteinfusion von MoHog1p mit dem Fluoreszenzprotein GFP konnte ein molekulares Werkzeug generiert werden, welches umfassende Analysen zur Aktivierung des HOG-Signalwegs erlaubt. Durch Expression des Fusionsproteins MoHog1p GFP in verschiedenen "loss of function"-Mutanten, bei denen einzelne Komponenten des HOG-Signalwegs inaktiviert sind, konnten so Untersuchungen zur Signalspezifität in diesem Signalweg durchgeführt werden. Unterstützt wurden Ergebnisse durch Untersuchungen des Primärmetabolismus von M. oryzae zur intrazellulären Akkumulation von Osmolyten nach Induktion mit diversen Stimuli. Zum einen konnte so zum ersten Mal gezeigt werden, dass Fludioxonil-Applikation analog zu hyperosmotischen Stimuli in M. oryzae zur Translokation von MoHog1p in den Zellkern führt, sowie die intrazelluläre Akkumulation des Osmolyts Arabitol bewirkt. Die molekularen Mechanismen der Perzeption und der Signaltransduktion bei Fludioxonil-Stress und hyperosmotischem Stress unterscheiden sich jedoch. Für eine Translokation von MoHog1p in den Zellkern und eine intrazelluläre Akkumulation von Arabitol bei Fludioxonil-Stress ist die Anwesenheit der Gruppe III HK MoHik1p essentiell. Für eine nukleäre Translokation von MoHog1p und eine adäquate intrazelluläre Akkumulation von Arabitol bei hyperosmotischem Schock hingegen nicht. Zum anderen konnte die Hypothese einer spezifischen Detektion von Salzstress bzw. Zuckerstress durch die HKs MoSln1p bzw. MoHik1p untersucht und schließlich spezifiziert werden: MoSln1p bzw. MoHik1p detektieren vermutlich nicht direkt Salz- oder Zuckerkonzentrationen, sondern wahrscheinlich sekundär unterschiedliche zelluläre Mechanismen oder Zustände, welche mit langfristigem Salz- oder Zuckerstress assoziiert sind oder dadurch jeweils spezifisch ausgelöst werden – nämlich möglicherweise Ionenstress, Hypoxie, oxidativen Stress, und/oder Änderungen im zellulären Redoxpotential. Weiterhin konnten potentiell unterschiedliche Aktivierungsmechanismen des HOG-Signalwegs postuliert werden, welche abhängig vom Stimulus entweder in einer direkten (hyperosmotischer Schock, Fludioxonil-Stress) oder verspäteten (Hypoxie / oxidativer Stress / Redox-Stress) Translokation von MoHog1p in den Zellkern resultieren.Summary The results of this work concerning the molecular mechanisms of the mode of action of the fungicide fludioxonil and on signaling functions of two-component hybrid histidine kinases (HKs) will make valuable contributions to the integrated plant protection management. It could be demonstrated that apart from genes known so far coding for proteins of the "High Osmolarity Glycerol" (HOG) signaling pathway, there are more components involved in fludioxonil susceptibility in the filamentous phytopathogenic fungus Magnaporthe oryzae. Homologous overexpression of the phosphatase MoPtp2p resulted in a fludioxonil resistant mutant with low MoHog1p phosphorylation, thus exhibiting a new molecular mechanism of fungicide resistance. Likewise, inactivation of MoPTP2 led to a fludioxonil hypersensitive phenotype. In conclusion MoPtp2p is involved in the signal transduction process in the HOG signaling pathway and is also crucial for the mode of action of the fungicide fludioxonil. The mitogen activated protein kinase MoHog1p, which is essential for osmoregulation, translocates from the cytosol into the nucleus upon pathway activation. Fusion of MoHog1p with the fluorescence protein GFP generated a molecular tool suitable for comprehensive visualization of HOG pathway activation. Expression of the fusion protein MoHog1p-GFP in various loss of function mutants of the HOG signaling pathway enabled sophisticated investigations of signaling specificity. These findings were supported by analyses of metabolomic changes upon application of varied stressors. Thus, it could be demonstrated that fludioxonil treatment leads to nuclear translocation of MoHog1, as well as to intracellular accumulation of the osmolyte arabitol – analogous to hyperosmotic shocks. However, the underlying molecular mechanisms for perception of fludioxonil stress and hyperosmotic stress differ. MoHik1p is essential for nuclear translocation of MoHog1p and intracellular accumulation of arabitol during fludioxonil stress, but not during hyperosmotic stress. Furthermore, the hypothesis of a specific detection of high molar salt or sugar stress by the HKs MoSln1p or MoHik1p could be investigated and finally specified: MoSln1p or MoHik1p do not detect specificly salt- or sugar concentrations, however, they probably detect secondary mechanisms or states associated with or initiated by long-term salt stress or long-term sugar stress – i.e. ion stress, hypoxia, oxidative stress and/or changing cellular redox potential. Finally two distinct HOG pathway activation mechanisms could be postulated, which result dependend on the stimulus in a rapid (hyperosmotic shock, fludioxonil stress) or delayed (hypoxia / oxidative stress / redox stress) nuclear translocation of MoHog1p