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Ferroptosis is a regulated form of cell death which is considered an oxidative iron-dependent process. The lipid hydroperoxidase glutathione peroxidase 4 prevents the iron (Fe2+)-dependent formation of toxic lipid reactive oxygen species. While emerging evidence indicates that inhibition of glutathione peroxidase 4 as a hallmark of ferroptosis in many cancer cell lines, the involvement of this biochemical pathway in neuronal death remains largely unclear. Here, we investigate, first whether the ferroptosis key players are involved in the neuronal cell death induced by erastin. The second objective was to examine whether there is a cross talk between ferroptosis and autophagy. The third main was to address neuron response to erastin, with a special focus on ferritin and nuclear receptor coactivator 4-mediated ferritinophagy. To test this in neurons, erastin (0.5–8 µM) was applied to hippocampal HT22 neurons for 16 hours. In addition, cells were cultured with the autophagy inhibitor, 3-methyladenin (10 mM) and/or ferroptosis inhibitors, ferrostatin 1 (10–20 µM) or deferoxamine (10–200 µM) before exposure to erastin. In this study, we demonstrated by immunofluorescence and western blot analysis, that erastin downregulates dramatically the expression of glutathione peroxidase 4, the sodium-independent cystine-glutamate antiporter and nuclear receptor coactivator 4. The protein levels of ferritin and mitochondrial ferritin in HT22 hippocampal neurons did not remarkably change following erastin treatment. In addition, we demonstrated that not only the ferroptosis inhibitor, ferrostatin1/deferoxamine abrogated the ferroptotic cell death induced by erastin in hippocampal HT22 neurons, but also the potent autophagy inhibitor, 3-methyladenin. We conclude that (1) erastin-induced ferroptosis in hippocampal HT22 neurons, despite reduced nuclear receptor coactivator 4 levels, (2) that either nuclear receptor coactivator 4-mediated ferritinophagy does not occur or is of secondary importance in this model, (3) that ferroptosis seems to share some features of the autophagic cell death process.
In the basal, proliferative layer of healthy skin, the mTOR complex 1 (mTORC1) is activated, thus regulating proliferation while preventing differentiation. When cells leave the proliferative, basal compartment, mTORC1 signaling is turned off, which allows differentiation. Under inflammatory conditions, this switch is hijacked by cytokines and prevents proper differentiation. It is currently unknown how mTORC1 is regulated to mediate these effects on keratinocyte differentiation. In other tissues, mTORC1 activity is controlled through various pathways via the tuberous sclerosis complex (TSC). Thus, we investigated whether the TS complex is regulated by proinflammatory cytokines and contributes to the pathogenesis of psoriasis. TNF-α as well as IL-1β induced the phosphorylation of TSC2, especially on S939 via the PI3-K/AKT and MAPK pathway. Surprisingly, increased TSC2 phosphorylation could not be detected in psoriasis patients. Instead, TSC2 was strongly downregulated in lesional psoriatic skin compared to non-lesional skin of the same patients or healthy skin. In vitro inflammatory cytokines induced dissociation of TSC2 from the lysosome, followed by destabilization of the TS complex and degradation. Thus, we assume that in psoriasis, inflammatory cytokines induce strong TSC2 phosphorylation, which in turn leads to its degradation. Consequently, chronic mTORC1 activity impairs ordered keratinocyte differentiation and contributes to the phenotypical changes seen in the psoriatic epidermis.
The culture of primary intestinal epithelia cells is not possible in a normal culture system. In 2009 a three-dimensional culture system of intestinal stem cells was established that shows many of the physiological features of the small intestine, such as crypt-villus structure, stem cell niche and all types of differentiated intestinal epithelial cells. These enteroids can be used to analyze biology of intestinal stem cells, gut homeostasis and the development of diseases. They also give the possibility to reduce animal numbers, as enteroids can be cryo-conserved and cultivated for many passages. To investigate the influence of genes such as NADPH oxidases on the gut homeostasis, transgenic approached are the method of choice. The generation of enteroids from knockout mice allows real-time observations of knockout effects. Often conditional knockout or overexpression strategies using inducible Cre recombinase are applied to avoid effects of adaption to the knockout. However, the Cre recombinase has many known caveats from unspecific binding and its endonuclease activity. In this study, we show that although NADPH oxidases are important for in vivo differentiation and proliferation of the intestine, their expression is drastically reduced in the organoid system. Activation of Cre recombinase by 4-hydroxy tamoxifen in freshly isolated enteroids, independently of floxed genes, leads to decreased diameter of organoids. This effect is concentration-dependent and is caused by reduced cell proliferation and induction of apoptosis and DNA damage. In contrast, constitutive expression of Cre has no impact on the enteroids. Therefore, reduction of tamoxifen concentration and treatment duration should be carefully titrated, and appropriate controls are necessary.
Koch, Vitali ; Abt, Julia Anna Walburga ; Grünewald, Leon David ; Eichler, Katrin ; D’Angelo, Tommaso ; Martin, Simon ; Albrecht, Moritz Hans Ernst ; Thalhammer, Axel ; Booz, Christian ; Yel, Ibrahim ; Simon Bernatz ; Mahmoudi, Scherwin ; Harth, Marc ; Derwich, Wojciech ; Vogl, Thomas J. ; Gray, Daphne ; Gruber-Rouh, Tatjana ; Jung, Georg
Purpose
To assess the diagnostic value of different imaging modalities in distinguishing systemic vasculitis from other internal and immunological diseases.
Methods
This retrospective study included 134 patients with suspected vasculitis who underwent ultrasound, magnetic resonance imaging (MRI), or 18F-fluorodeoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT) between 01/2010 and 01/2019, finally consisting of 70 individuals with vasculitis. The main study parameter was the confirmation of the diagnosis using one of the three different imaging modalities, with the adjudicated clinical and histopathological diagnosis as the gold standard. A secondary parameter was the morphological appearance of the vessel affected by vasculitis.
Results
Patients with systemic vasculitis had myriad clinical manifestations with joint pain as the most common symptom. We found significant correlations between different imaging findings suggestive of vasculitis and the final adjudicated clinical diagnosis. In this context, on MRI, vessel wall thickening, edema, and diameter differed significantly between vasculitis and non-vasculitis groups (p < 0.05). Ultrasound revealed different findings that may serve as red flags in identifying patients with vasculitis, such as vascular occlusion or halo sign (p = 0.02 vs. non-vasculitis group). Interestingly, comparing maximal standardized uptake values from PET/CT examinations with vessel wall thickening or vessel diameter did not result in significant differences (p > 0.05).
Conclusions
We observed significant correlations between different imaging findings suggestive of vasculitis on ultrasound or MRI and the final adjudicated diagnosis. While ultrasound and MRI were considered suitable imaging methods for detecting and discriminating typical vascular changes, 18F-FDG PET/CT requires careful timing and patient selection given its moderate diagnostic accuracy.
de Vries, Jean-Paul P. M. ; Zuidema, Roy ; Bicknell, Colin D. ; Fisher, Robert ; Gargiulo, Mauro ; Louis, Nicolas ; Oikonomou, Kyriakos ; Pratesi, Giovanni ; Reijnen, Michel ; Reyes Valdivia, Andrés ; Riambau, Vincent ; Saucy, François
Purpose:
The purpose of the study was to provide a consensus definition of the infrarenal sealing zone and develop an algorithm to determine when and if adjunctive procedure(s) or reintervention should be considered in managing patients undergoing endovascular aortic repair (EVAR) for infrarenal abdominal aortic aneurysm (AAA).
Methods:
A European Advisory Board (AB), made up of 11 vascular surgeons with expertise in EVAR for AAA, was assembled to share their opinion regarding the definition of preoperative and postoperative infrarenal sealing zone. Information on their current clinical practice and level of agreement on proposed reintervention paths was used to develop an algorithm. The process included 2 virtual meetings and 2 rounds of online surveys completed by the AB (Delphi method). Consensus was defined as reached when ≥ 8 of 11 (73%) respondents agreed or were neutral.
Results:
The AB reached complete consensus on definitions and measurement of the pre-EVAR target anticipated sealing zone (TASZ) and the post-EVAR real achieved sealing zone (RASZ), namely, the shortest length between the proximal and distal reference points as defined by the AB, in case of patients with challenging anatomies. Also, agreement was achieved on a list of 4 anatomic parameters and 3 prosthesis-/procedure-related parameters, considered to have the most significant impact on preoperative and postoperative sealing zones. Furthermore, the agreement was reached that in the presence of visible neck-related complications, both adjunctive procedure(s) and reintervention should be contemplated (100% consensus). In addition, adjunctive procedure(s) or reintervention can be considered in the following cases (% consensus): insufficient sealing zone on completion imaging (91%) or on the first postoperative computed tomography (CT) scan (91%), suboptimal sealing zone on completion imaging (73%) or postoperative CT scan (82%), and negative evolution of the actual sealing zone over time (91%), even in the absence of visible complications.
Conclusions:
AB members agreed on definitions of the pre- and post-EVAR infrarenal sealing zone, as well as factors of influence. Furthermore, a clinical decision algorithm was proposed to determine the timing and necessity of adjunctive procedure(s) and reinterventions.
Kattih, Badder ; Carstens, Daniel C. ; Böckling, Felicitas Juana ; Rasper, Tina ; Pergola, Graziella ; Dimmeler, Stefanie ; Vasa‐Nicotera, Mariuca ; Zeiher, Andreas M. ; Mas-Peiro, Silvia
Background
Musclin is an activity‐stimulated and cardioprotective myokine that attenuates pathological cardiac remodeling. Musclin deficiency, in turn, results in reduced physical endurance. The aim of this study was to assess the prognostic value of circulating musclin as a novel, putative biomarker to identify patients undergoing transcatheter aortic valve implantation (TAVI) who are at a higher risk of death.
Methods and Results
In this study, we measured systemic musclin levels in 368 patients undergoing TAVI who were at low to intermediate clinical risk (median EuroSCORE [European System for Cardiac Operative Risk Evaluation] II: 3.5; quartile 1–quartile, 2.2%–5.3%), whereby 209 (56.8%) patients were at low and 159 (43.2%) were at intermediate risk. Median preprocedural musclin levels were 2.7 ng/mL (quartile 1–quartile 3, 1.5–4.6 ng/mL). Musclin levels were dichotomized in low (<2.862 ng/mL, n=199 [54.1%]) or high (≥ 2.862 ng/mL, n=169 [45.9%]) groups using cutoff values determined by classification and regression tree analysis. The primary end point was 1‐year overall survival. Patients with low circulating musclin levels exhibited a significantly higher prevalence of frailty, low albumin values, hypertension, and history of stroke as well as higher N‐terminal pro‐B‐type natriuretic peptide. Low musclin levels significantly predicted risk of death in univariable (hazard ratio, 1.81; 95% CI, 1.00–3.53 [P=0.049]) and multivariable (adjusted hazard ratio, 2.45; 95% CI, 1.06–5.69 [P=0.037]) Cox regression analyses. Additionally, low musclin levels in combination with conventional EuroSCORE II suggested improved risk stratification in patients undergoing TAVI who were at low to intermediate clinical risk into subgroups with reduced 1‐year survival rates by log‐rank test (P for trend=0.003).
Conclusions
Circulating musclin is an independent predictor of 1‐year overall survival in patients undergoing TAVI. Combined with EuroSCORE II, circulating musclin might help to improve prediction of mortality in patients undergoing TAVI who are at low to intermediate clinical risk.
Infection with the SARS (Severe Acute Respiratory Syndrome)-associated coronavirus results in respiratory failure probably by immunological mechanisms in 10% of patients. Laboratory markers that predict subsequent respiratory failure would therefore be useful in patient management.
We describe the clinical course, hematologic parameters, lymphocyte subpopulations and markers of inflammation in two patients with SARS, i.e., one man with diabetes mellitus and one pregnant woman, infected by the same viral isolate.
The patient with underlying diabetes mellitus developed respiratory failure after admission in the second week of the illness while the second patient developed only a mild disease without respiratory failure. Subsequent respiratory dysfunction was associated with lown umbers of Natural Killer (NK) cells at presentation and elevated CRP levels during the illness.
NK cells and CRP levels at the end of the first week of the disease might be related to subsequent respiratory dysfunction and may link underlying conditions to disease severity.
Seredinski, Sandra ; Boos, Frederike ; Günther, Stefan ; Oo, James A. ; Warwick, Timothy ; Izquierdo Ponce, Judit ; Lillich, Felix F. ; Proschak, Ewgenij ; Knapp, Stefan ; Gilsbach, Ralf ; Pflüger-Müller, Beatrice ; Brandes, Ralf ; Leisegang, Matthias
The transcription factor hypoxia-inducible factor 1 (HIF1) is an important driver of cancer and is therefore an attractive drug target. Acriflavine (ACF) has been suggested to inhibit HIF1, but its mechanism of action is unknown. Here we investigated the interaction of ACF with DNA and long non-coding RNAs (lncRNAs) and its function in human endothelial cells. ACF promoted apoptosis and reduced proliferation, network formation, and angiogenic capacity. It also induced changes in gene expression, as determined by RNA sequencing (RNA-seq), which could not be attributed to specific inhibition of HIF1. A similar response was observed in murine lung endothelial cells. Although ACF increased and decreased a similar number of protein-coding genes, lncRNAs were preferentially upregulated under normoxic and hypoxic conditions. An assay for transposase accessibility with subsequent DNA sequencing (ATAC-seq) demonstrated that ACF induced strong changes in chromatin accessibility at lncRNA promoters. Immunofluorescence showed displacement of DNA:RNA hybrids. Such effects might be due to ACF-mediated topoisomerase inhibition, which was indeed the case, as reflected by DNA unwinding assays. Comparison with other acridine derivatives and topoisomerase inhibitors suggested that the specific function of ACF is an effect of acridinium-class compounds. This study demonstrates that ACF inhibits topoisomerases rather than HIF specifically and that it elicits a unique expression response of lncRNAs.
Lambregts, Doenja M. J. ; Bogveradze, Nino ; Blomqvist, Lennart K. ; Fokas, Emmanouil ; Garcia-Aguilar, Julio ; Glimelius, Bengt ; Gollub, Marcus ; Konishi, Tsuyoshi ; Marijnen, Corrie A. M. ; Nagtegaal, Iris D. ; Nilsson, Per J. ; Perez, Rodrigo O. ; Snaebjornsson, Petur ; Taylor, Stuart A. ; Tolan, Damian J. M. ; Valentini, Vincenzo ; West, Nicholas P. ; Wolthuis, Albert ; Lahaye, Max J. ; Maas, Monique ; Beets, Geerard ; Beets-Tan, Regina G. H.
Objectives
To identify the main problem areas in the applicability of the current TNM staging system (8th ed.) for the radiological staging and reporting of rectal cancer and provide practice recommendations on how to handle them.
Methods
A global case-based online survey was conducted including 41 image-based rectal cancer cases focusing on various items included in the TNM system. Cases reaching < 80% agreement among survey respondents were identified as problem areas and discussed among an international expert panel, including 5 radiologists, 6 colorectal surgeons, 4 radiation oncologists, and 3 pathologists.
Results
Three hundred twenty-one respondents (from 32 countries) completed the survey. Sixteen problem areas were identified, related to cT staging in low-rectal cancers, definitions for cT4b and cM1a disease, definitions for mesorectal fascia (MRF) involvement, evaluation of lymph nodes versus tumor deposits, and staging of lateral lymph nodes. The expert panel recommended strategies on how to handle these, including advice on cT-stage categorization in case of involvement of different layers of the anal canal, specifications on which structures to include in the definition of cT4b disease, how to define MRF involvement by the primary tumor and other tumor-bearing structures, how to differentiate and report lymph nodes and tumor deposits on MRI, and how to anatomically localize and stage lateral lymph nodes.
Conclusions
The recommendations derived from this global survey and expert panel discussion may serve as a practice guide and support tool for radiologists (and other clinicians) involved in the staging of rectal cancer and may contribute to improved consistency in radiological staging and reporting.
While in large clinical laboratories the implementation of total laboratory automation is continuously proceeding, this concept is mostly not suitable for small- and middle-sized laboratories or for testing laboratories of blood donation services due to costs and required space. For these facilities, however, a rational level of automation can be achieved by the installation of stand-alone work cells for pre-analytical and selected analytical processes. In this review, the features of some automated pre-analytical sample processing systems and automated systems for the serological testing for infectious diseases are described exemplarily and compared with each other. The major advantages of automated systems, compared to a solely manual workflow, are described. Essential factors which have to be considered for making the choice for an appropriate automated system are pointed out.
Koll, Florestan ; Meisenzahl, Eva ; Haller, Bernhard ; Maisch, Philipp ; Kirchhoff, Florian ; Horn, Thomas ; Gschwend, Jürgen E. ; Schmid, Sebastian C.
Purpose:
Discordance between pre-operative biopsy and final pathology for Upper Tract Urothelial Carcinoma (UTUC) is high and optimal management remains controversial. The aim of this study is to evaluate the accuracy of pre-operative biopsy, to identify prognostic factors and to evaluate the effect of adjuvant chemotherapy on survival and oncologic outcome in UTUC.
Methods:
We analyzed records of patients receiving surgical treatment for UTUC. Pathology of pre-operative biopsy was compared to surgical specimen. We used Kaplan-Meier method to estimate survival probabilities and Cox's proportional hazards models to estimate the association between covariates and event times. Primary endpoint was overall survival (OS). A matched-pair analysis was performed to evaluate the effect of adjuvant chemotherapy.
Results:
151 patients underwent surgical treatment (28% open, 36% laparoscopic, 17% robotic radical nephroureterectomy; 14% segmental ureteral resections and 5% palliative nephrectomy) for UTUC and were included in the analysis. Upstaging from <pT1 in endoscopic biopsy to ≥pT1 in final pathology occurred in 61% of patients and upgrading from low-grade to high-grade occurred in 30% of patients. Five-year OS was 59.5%. In the univariate Cox-regression model pathological stage, grade, lymphovascular invasion and positive surgical margins were associated with OS. Matched pair analysis for stage (<pT3; ≥pT3; pN+) and age revealed a significant survival benefit for adjuvant chemotherapy (HR 0.40, 0.14–0.77, p < 0.018) in this cohort.
Conclusion:
UTUC is often underestimated in pre-operative biopsy, and it is associated with significant mortality. Pathological stage and grade, lymphovascular invasion and lymph node metastases are predictors of oncologic outcome and survival.
Congenital diaphragmatic hernia (CDH) is a relatively common and life-threatening birth defect, characterized by an abnormal opening in the primordial diaphragm that interferes with normal lung development. As a result, CDH is accompanied by immature and hypoplastic lungs, being the leading cause of morbidity and mortality in patients with this condition. In recent decades, various animal models have contributed novel insights into the pathogenic mechanisms underlying CDH and associated pulmonary hypoplasia. In particular, the generation of genetically modified mouse models, which show both diaphragm and lung abnormalities, has resulted in the discovery of multiple genes and signaling pathways involved in the pathogenesis of CDH. This article aims to offer an up-to-date overview on CDH-implicated transcription factors, molecules regulating cell migration and signal transduction as well as components contributing to the formation of extracellular matrix, whilst also discussing the significance of these genetic models for studying altered lung development with regard to the human situation.
Trojan, Jörg ; Hoffmeister, Albrecht ; Neu, Bruno ; Kasper-Virchow, Stefan ; Dechêne, Alexander ; Jürgensen, Christian ; Schirra, Jörg ; Jakobs, Ralf ; Palmer, Dan ; Selbo, Pål k. ; Olivecrona, Hans ; Finnesand, Lena ; Høgset, Anders ; Walday, Per ; Sturgess, Richard
Background
Photochemical internalization (PCI) is a novel technology for light-induced enhancement of the local therapeutic effect of cancer drugs, utilizing a specially designed photosensitizing molecule (fimaporfin). The photosensitizing molecules are trapped in endosomes along with macromolecules or drugs. Photoactivation of fimaporfin disrupts the endosomal membranes so that drug molecules are released from endosomes inside cells and can reach their therapeutic target in the cell cytosol or nucleus. Compared with photodynamic therapy, the main cytotoxic effect with PCI is disruption of the endosomal membrane resulting in delivery of chemotherapy drug, and not to the photochemical reactions per se. In this study we investigated the effect of PCI with gemcitabine in patients with inoperable perihilar cholangiocarcinoma (CCA).
Methods
The in vitro cytotoxic effect of PCI with gemcitabine was studied on two CCA-derived cell lines. In a fimaporfin dose-escalation phase I clinical study, we administered PCI with gemcitabine in patients with perihilar CCA (n = 16) to establish a safe and tolerable fimaporfin dose and to get early signals of efficacy. The patients enrolled in the study had tumors in which the whole length of the tumor could be illuminated from the inside of the bile duct, using an optical fiber inserted via an endoscope (Fig. 1). Fimaporfin was administered intravenously at day 0; gemcitabine (i.v.) and intraluminal biliary endoscopic laser light application on day 4; followed by standard gemcitabine/cisplatin chemotherapy.
Results
Preclinical experiments showed that PCI enhanced the effect of gemcitabine. In patients with CCA, PCI with gemcitabine was well tolerated with no dose-limiting toxicities, and no unexpected safety signals. Disease control was achieved in 10 of 11 evaluable patients, with a clearly superior effect in the two highest dose groups. The objective response rate (ORR) was 42%, including two complete responses, while ORR at the highest dose was 60%. Progression-free survival at 6 months was 75%, and median overall survival (mOS) was 15.4 months, with 22.8 months at the highest fimaporfin dose.
Conclusion
Photochemical internalization with gemcitabine was found to be safe and resulted in encouraging response and survival rates in patients with unresectable perihilar CCA.
Microstates sind kurzzeitig andauernde, wiederkehrende elektrische Potentialfelder über dem Kortex. Ein Großteil der Signalvarianz des
Elektroenzephalogramms (EEG) wird durch vier repräsentative räumliche Potentialverteilungen (Topographien) abgedeckt, welche bereits im Wachzustand und im Schlaf identifiziert wurden und kanonisch als Karten A-D bezeichnet werden. Microstates wurden in den vergangenen Jahren vor allem im Ruhe-Wach-EEG untersucht, über andere Vigilanzzustände hingegen wissen wir bisher wenig. Klassischerweise analysieren wir verschiedene Vigilanzzustände im Elektroenzephalogramm anhand von Frequenzen und Graphoelementen, die Microstate-Analyse hingegen betrachtet in erster Linie die räumliche Verteilung des kortikalen Potentials zu einem jeweiligen Zeitpunkt.
Die vorliegende Studie hatte zum Ziel, die zeitliche Abfolge von Microstates im Wachzustand und im Schlaf zu charakterisieren. Mittels informationstheoretischer Ansätze können die dynamischen Eigenschaften der Microstate-Sequenz direkt mit den frequenzbasierten Eigenschaften des zugrundeliegenden EEG verglichen werden. Es wurden die Ruhe-Wach- und Schlafdaten von 32 gesunden Probanden analysiert. Hierbei fand sich eine Zunahme der mittleren Microstate-Dauer und der Relaxationszeit der Übergangsmatrix, was langsamere Dynamiken im Schlaf anzeigt. Erstaunlicherweise konnte im Tiefschlaf mehr als die Hälfte der Sequenzen nicht von einem simplen Markov-Modell unterschieden werden, was für eine Abnahme der Komplexität der Microstate-Sequenzen spricht. Die Entropierate der untersuchten Sequenzen nahm mit zunehmender Schlaftiefe ab, was weniger
Zufall bzw. eine größere Vorhersagbarkeit innerhalb der Sequenzen bedeutet.
Darüberhinaus konnte gezeigt werden, dass Microstates immer dann periodisch auftreten, wenn das zugrundeliegende EEG eine dominante Grundfrequenz aufweist, sodass oszillatorische Hirnaktivität auch auf der Microstate-Ebene verfolgbar ist. Hierdurch ist es möglich, physiologische Vigilanzzustände quantitativ voneinander zu unterscheiden.
Interpretiert man Microstates als Korrelate neuronaler Netzwerke, scheinen im Schlaf dieselben oder ähnliche Netzwerke aktiviert zu werden wie im Wachzustand, allerdings mit zunehmender Schlaftiefe langsamer und auf eine weniger komplexe Art und Weise.
Wissenschaftsbasierte und verständliche Gesundheitsinformationen sind ein Kernelement der Evidenzbasierten Medizin und von Public Health. Ziel ist es, informierte Entscheidungen zu ermöglichen, die auf realistischen Einschätzungen von Gesundheitsrisiken sowie von Nutzen und Schaden möglicher Interventionen beruhen. In Deutschland wurden während der COVID-19-Pandemie die Standards für eine evidenzbasierte Risikokommunikation wenig beachtet. Häufig war die öffentliche Berichterstattung einseitig, unvollständig und missverständlich. Bedrohungsszenarien haben emotionalen Stress und unnötige Angst ausgelöst. Eine systematische und umfassende Aufarbeitung der Pandemiemaßnahmen ist auch in Deutschland dringlich geboten. Dabei müsste eine kritisch-konstruktive Analyse der medialen Risikokommunikation von Expert*innen, Politiker*innen und Medien ein zentrales Element der Aufarbeitung sein. Die Ergebnisse sollen helfen, aus der vergangenen Pandemie zu lernen, um für künftige Krisen besser vorbereitet zu sein.
Patyna, Ralf Sammy ; Eckes, Timon Sebastian ; Koch, Benjamin Florian ; Sudowe, Stephan ; Oftring, Anke ; Kohmer, Niko ; Rabenau, Holger ; Ciesek, Sandra ; Avaniadi, Parthenopi Despina ; Steiner, Rahel Elisabeth ; Hauser, Ingeborg A. ; Pfeilschifter, Josef ; Betz, Christoph
The long-term effect of protection by two doses of SARS-CoV-2 vaccination in patients receiving chronic intermittent hemodialysis (CIHD) is an urging question. We investigated the humoral and cellular immune response of 42 CIHD patients who had received two doses of SARS-CoV-2 vaccine, and again after a booster vaccine with mRNA-1273 six months later. We measured antibody levels and SARS-CoV-2-specific surrogate neutralizing antibodies (SNA). Functional T cell immune response to vaccination was assessed by quantifying interferon-γ (IFN-γ) and IL-2 secreting T cells specific for SARS-CoV-2 using an ELISpot assay. Our data reveal a moderate immune response after the second dose of vaccination, with significantly decreasing SARS-CoV-2-specific antibody levels and less than half of the study group showed neutralizing antibodies six months afterwards. Booster vaccines increased the humoral response dramatically and led to a response rate of 89.2% for antibody levels and a response rate of 94.6% for SNA. Measurement in a no response/low response (NR/LR) subgroup of our cohort, which differed from the whole group in age and rate of immunosuppressive drugs, indicated failure of a corresponding T cell response after the booster vaccine. We strongly argue in favor of a regular testing of surrogate neutralizing antibodies and consecutive booster vaccinations for CIHD patients to provide a stronger and persistent immunity.
Fanelli, Giuseppe ; Franke, Barbara ; Fabbri, Chiara ; Werme, Josefin ; Erdogan, Izel ; De Witte, Ward ; Poelmans, Geert ; Ruisch, I. Hyun ; Reus, Lianne Maria ; Gils, Veerle van ; Jansen, Willemijn J. ; Vos, Stephanie J.B. ; Alam, Kazi Asraful ; Martinez, Aurora ; Haavik, Jan ; Wimberley, Theresa ; Dalsgaard, Søren ; Fóthi, Ábel ; Barta, Csaba ; Fernández-Aranda, Fernando ; Jiménez-Murcia, Susana ; Berkel, Simone ; Matura, Silke ; Salas-Salvadó, Jordi ; Arenella, Martina ; Serretti, Alessandro ; Mota, Nina Roth ; Bralten, Janita
Abstract
The co-occurrence of insulin resistance (IR)-related metabolic conditions with neuropsychiatric disorders is a complex public health challenge. Evidence of the genetic links between these phenotypes is emerging, but little is currently known about the genomic regions and biological functions that are involved. To address this, we performed Local Analysis of [co]Variant Association (LAVA) using large-scale (N=9,725-933,970) genome-wide association studies (GWASs) results for three IR-related conditions (type 2 diabetes mellitus, obesity, and metabolic syndrome) and nine neuropsychiatric disorders. Subsequently, positional and expression quantitative trait locus (eQTL)-based gene mapping and downstream functional genomic analyses were performed on the significant loci. Patterns of negative and positive local genetic correlations (|rg|=0.21-1, pFDR<0.05) were identified at 109 unique genomic regions across all phenotype pairs. Local correlations emerged even in the absence of global genetic correlations between IR-related conditions and Alzheimer’s disease, bipolar disorder, and Tourette’s syndrome. Genes mapped to the correlated regions showed enrichment in biological pathways integral to immune-inflammatory function, vesicle trafficking, insulin signalling, oxygen transport, and lipid metabolism. Colocalisation analyses further prioritised 10 genetically correlated regions for likely harbouring shared causal variants, displaying high deleterious or regulatory potential. These variants were found within or in close proximity to genes, such as SLC39A8 and HLA-DRB1, that can be targeted by supplements and already known drugs, including omega-3/6 fatty acids, immunomodulatory, antihypertensive, and cholesterol-lowering drugs. Overall, our findings underscore the complex genetic landscape of IR-neuropsychiatric multimorbidity, advocating for an integrated disease model and offering novel insights for research and treatment strategies in this domain.
Highlights
Local genetic correlations found even in the absence of global correlations.
Both positive and negative local correlations found for IR-neuropsychiatric pairs.
Enrichment for immune, and insulin signalling pathways, among others.
Pinpointed shared likely causal variants within 10 genomic regions.
Identified therapeutic targets, e.g., SLC39A8 and HLA-DRB1, for drug repurposing.
Die Nukleinsäure-Amplifikations Testung (NAT) von Blutprodukten wurde Mitte der 90er Jahre von europäischen Plasma verarbeitenden Firmen und großen deutschen Blutspendediensten entwickelt. Primäres Ziel war eine verbesserte Sicherheit von Blutprodukten, indem das so genannte diagnostische Fenster nach einer Virusinfektion bis zum ersten Nachweis von Antikörpern so weit wie möglich geschlossen werden sollte. Bei einer qualitätsgerechten PCR kommen bereits der Probenentnahme, dem Probentransport sowie der Probenlagerung große Bedeutung zu, da vermieden werden muß, daß es durch ungeeignete Antikoagulanzien oder Entnahmetechniken zu einem Sensitivitätsverlust kommt oder daß Kontaminationen falsch positive Ergebnisse hervorrufen. Wird ein Pooling von Proben durchgeführt, ergibt sich ein Verdünnungsfaktor, weshalb darauf zu achten ist, dass gegebenenfalls nachfolgende Anreicherungsschritte für Viren, wie z.B. eine Zentrifugation, implementiert werden. Der Gesamtprozeß von Pooling und Virusanreicherung ist ebenso wie die Probenvorbereitung durch geeignete Maßnahmen zu validieren und durch Qualitätssicherungsmaßnahmen zu flankieren. Die in der Extraktion der viralen Nukleinsäuren verwendeten Reagenzien sollten im Laboralltag möglichst einfach zu handhaben sein, keine Gefährdung des Laborpersonals darstellen und die Virus-Nukleinsäure gleichzeitig mit höchster Effizienz freisetzen und in sehr hoher Reinheit für die anschließende Amplifikation bereitstellen. Qualitätssicherungmaßnahmen sollen hier sowohl die geforderte Effizienz des Prozesses sichern als auch verhindern, daß es in dieser kritischen Phase zu Kontaminationen kommt. Zur Amplifikation stehen verschiedene Methoden zur Verfügung, wobei die PCR, insbesondere bei inhouse-Systemen, die weiteste Verbreitung gefunden hat. Der Prozeß der Amplifikation sollte möglichst im geschlossenen System erfolgen, wie dies z.B. in Real-time PCR-Systemen die Regel ist, ohne daß das Reaktionsgefäß während oder nach dem Amplifikationsprozeß geöffnet werden muß. Dies gewährleistet eine hohe Sicherheit vor Kontaminationen durch freigesetzte Amplifikate. Im Blutspendewesen ist es von höchster Bedeutung, daß negative Ergebnisse tatsächlich negative Blutspenden anzeigen. Interne Kontrollen, die eine korrekte Funktionsweise jeder individuellen PCR signalisieren, sollten deshalb in jeder Reaktion mitgeführt werden. Neben internen Kontrollen sind externe Negativ- und Positiv-Kontrollen mitzuführen, um falsch positive Reaktionen nachzuweisen bzw. auch die vor der PCR liegenden Prozesse wie Virusanreicherung und Extraktion zu überwachen. Alle Prozesse sind nach den von den Behörden festgelegten Kriterien durchgängig zu validieren, und es ist routinemäßig an externen Qualitätskontrollmaßnahmen (Ringversuchen) teilzunehmen.
Feist, Eugen ; Baraliakos, Xenofon ; Behrens, Frank ; Thaçi, Diamant ; Klopsch, Thilo ; Plenske, Anja ; Blindzellner, Lisa K. ; Klaus, Pascal ; Meng, Thomas ; Löschmann, Peter-Andreas
Feist, Eugen ; Baraliakos, Xenofon ; Behrens, Frank ; Thaçi, Diamant ; Klopsch, Thilo ; Plenske, Anja ; Blindzellner, Lisa K. ; Klaus, Pascal ; Meng, Thomas ; Löschmann, Peter-Andreas
Background: For rheumatoid arthritis (RA), the treat-to-target concept suggests attaining remission or at least low disease activity (LDA) after 12 weeks.
Objectives: This German, prospective, multicenter, non-interventional study aimed to determine the proportion of patients with RA who achieved their treat-to-target aim after 12 and 24 weeks of etanercept (ETN) treatment in a real-life setting, as opposed to patients achieving their therapeutic target at a later timepoint (week 36 or 52).
Methods: A total of 824 adults with a confirmed diagnosis of RA without prior ETN treatment were included. Remission and LDA were defined as DAS28 < 2.6 and DAS28 ≤ 3.2, respectively.
Results: The proportion of patients achieving remission was 24% at week 12 and 31% at week 24. The proportion of patients achieving LDA was 39% at week 12 and 45% at week 24. The proportion of patients achieving remission or LDA further increased beyond week 24 up to week 52. Improvement in pain and reduction in concomitant glucocorticoid treatment were observed. Improvements in patient-reported outcomes were also seen in patients who did not reach remission or LDA. No new safety signals were detected.
Conclusions: A considerable proportion of patients with RA attained the target of remission or LDA after 12 weeks of ETN treatment. Even beyond that timepoint, the proportion of patients achieving treatment targets continued to increase up to week 52.
Trial Registration
ClinicalTrials.gov Identifier: NCT02486302.
Plain Language Summary
Physicians measure response to treatment of rheumatoid arthritis using a disease activity score (DAS28). People with a DAS28 of less than 2.6 have very few to no symptoms (also called remission). People with a DAS28 of 3.2 or less, called low disease activity, may experience mild symptoms. When people do not respond to treatment after 12 weeks, it is usually recommended to prescribe a different treatment. Researchers do not know how many people who do not respond after 12 weeks would respond if treatment were continued. A total of 824 German people with rheumatoid arthritis who received a drug called etanercept for up to 52 weeks took part in this study. Researchers wanted to know how many people had remission or low disease activity after 12 weeks and 24 weeks of treatment.
After 12 weeks, 24 in 100 people had remission; this increased to 31 in 100 people after 24 weeks. Thirty-nine in 100 people had low disease activity after 12 weeks; this increased to 45 in 100 people after 24 weeks. The number of people with remission or low disease activity increased with longer treatment (up to 52 weeks). People needed less additional treatment with a type of drug called glucocorticoids. The people in this study experienced side effects that were similar to those reported by people who took etanercept in previous studies.
The researchers concluded that a considerable proportion of people responded to treatment with etanercept after 12 weeks. This proportion increased when treatment was continued for longer than 12 weeks.
Porous tantalum trabecular metal biomaterial has a similar structure to trabecular bone, and was recently added to titanium dental implants as a surface enhancement. The purpose of this prospective pilot study was to describe 5-year survival results and crestal bone level changes around immediately-provisionalized Trabecular Metal Dental Implants. Eligible patients were adults in need of ≥1 implants in the posterior jaw. A non-occluding single acrylic provisional crown was in place for up to 14 days before final restoration. Clinical evaluations with radiographs were conducted at each follow-up visit (1 month, 3 months, 6 months, and 1 to 5 years). The primary endpoint was implant survival, characterized using the Kaplan-Meier method. The secondary endpoint was changes in crestal bone level, evaluated using a paired t-test to compare mean crestal bone levels between the baseline, 6-month, and annual follow-up values. In total, 30 patients (37 implants) were treated. Mean patient age was 45.5 years, and 63% were female. There was one implant failure; cumulative survival at 5 years was 97.2%. After the initial bone loss of 0.40 mm in the first 6 months, there were no statistically significant changes in crestal bone level over time up to 5 years of follow-up.
Trebicka, Jonel ; Gu, Wenyi ; De Ledinghen, Victor ; Aubé, Christophe ; Krag, Aleksander ; Praktiknjo, Michael ; Castera, Laurent ; Dumortier, Jerome ; Bauer, David Josef Maria ; Friedrich-Rust, Mireen ; Pol, Stanislas ; Grgurevic, Ivica ; Zheng, Rongqin ; Francque, Sven ; Gottfriedovà, Halima ; Mustapic, Sanda ; Sporea, Ioan ; Berzigotti, Annalisa ; Uschner, Frank Erhard ; Simbrunner, Benedikt Michael ; Ronot, Maxime ; Cassinotto, Christophe ; Kjaergaard, Maria ; Andrade, Filipe ; Schulz, Martin ; Semmler, Georg ; Tjesic Drinkovic, Ida ; Chang, Johannes ; Brol, Maximilian ; Rautou, Pierre Emmanuel ; Vanwolleghem, Thomas ; Straßburg, Christian P. ; Boursier, Jerome ; Ferstl, Philip ; Rasmussen, Ditlev Nytoft ; Reiberger, Thomas ; Vilgrain, Valerie ; Guibal, Aymeric ; Guillaud, Olivier ; Zeuzem, Stefan ; Vassord, Camille ; Lu, Xue ; Vonghia, Luisa ; Senkerikova, Renata ; Popescu, Alina ; Margini, Cristina ; Wang, Wenping ; Thiele, Maja ; Jansen, Chrisitan
Objective: Liver stiffness measurement (LSM) is a tool used to screen for significant fibrosis and portal hypertension. The aim of this retrospective multicentre study was to develop an easy tool using LSM for clinical outcomes in advanced chronic liver disease (ACLD) patients.
Design: This international multicentre cohort study included a derivation ACLD patient cohort with valid two-dimensional shear wave elastography (2D-SWE) results. Clinical and laboratory parameters at baseline and during follow-up were recorded. LSM by transient elastography (TE) was also recorded if available. The primary outcome was overall mortality. The secondary outcome was the development of first/further decompensation.
Results: After screening 2148 patients (16 centres), 1827 patients (55 years, 62.4% men) were included in the 2D-SWE cohort, with median liver SWE (L-SWE) 11.8 kPa and a model for end stage liver disease (MELD) score of 8. Combination of MELD score and L-SWE predict independently of mortality (AUC 0.8). L-SWE cut-off at ≥20 kPa combined with MELD ≥10 could stratify the risk of mortality and first/further decompensation in ACLD patients. The 2-year mortality and decompensation rates were 36.9% and 61.8%, respectively, in the 305 (18.3%) high-risk patients (with L-SWE ≥20 kPa and MELD ≥10), while in the 944 (56.6%) low-risk patients, these were 1.1% and 3.5%, respectively. Importantly, this M10LS20 algorithm was validated by TE-based LSM and in an additional cohort of 119 patients with valid point shear SWE-LSM.
Conclusion: The M10LS20 algorithm allows risk stratification of patients with ACLD. Patients with L-SWE ≥20 kPa and MELD ≥10 should be followed closely and receive intensified care, while patients with low risk may be managed at longer intervals.
Rationale: Studies have suggested mental health improvements following the use of the psychotropic plant concoction ayahuasca in non-clinical and clinical samples.
Objectives: The present observational study assessed depressive symptomatology in 20 clinically depressed patients (symptom score > 13 on the Beck’s Depression Inventory) before attendance of an ayahuasca ceremony and 1 month and 1 year after. Secondary measures included ratings of altered states of consciousness and ego dissolution during the ayahuasca ceremony as well as global measures of mindfulness, satisfaction with life, depression, anxiety, and stress.
Results: Twenty participants completed baseline and 1-day follow-up, 19 completed measures at 1-month follow-up, and 17 completed measures at 1-year follow-up. BDI scores reduced from baseline (M = 22.7) to all post-ceremony measures (Ms 11.45, 12.89, and 8.88, for 1-day, 1-month, and 1-year follow-up, respectively). After 1 day, 12/20 participants were in remission (BDI < 13). Remission rates after 1 month and 1 year were 13/19 and 12/17, respectively. Three participants remained mildly depressed (BDI 14–19) at the 1-month and 1-year follow-up. Two participants did not respond and remained at a moderate/severe level of depression at 1-year follow-up. Reductions on the secondary mental health measures and increases in mindfulness and satisfaction with life were found up to 1 year post-ceremony. Improvements in clinical depression and mental health correlated with levels of experienced ego dissolution and oceanic boundlessness during the ceremony up to 1 month after the ceremony. Engagement in additional mental health treatments or use of another psychedelic during study participation may have contributed to improved mental health ratings at 1-year follow-up.
Conclusion: Ayahuasca produces long-term mental health improvements in clinically depressed patients, which highlights its therapeutic potential.
Oxytocin, welches primär als Hormon bekannt ist, beeinflusst als Neuromodulator viele kognitive Prozesse, die an sozialem Verhalten, wie Sprache, beteiligt sind. Einerseits verändert es akustische Merkmale von gesprochener Sprache, andererseits erleichtert es auf perzeptueller Ebene die Emotionserkennung in der Sprachwahrnehmung und Körpersprache. Bislang war nicht bekannt, wie Oxytocin Hirnaktivität während des Sprechens verändert. Wir hypothetisierten, dass dieser Neuromodulator ähnlich wie Dopamin kortiko-basale Schaltkreise bahnen könnte.
Wir führten eine doppelt-verblindete Verhaltens- und funktionelle Kernspintomographiestudie durch, in der 52 gesunde Probanden an zwei getrennten Untersuchungsterminen entweder intranasales Oxytocin oder ein Placebo erhielten. Die Teilnehmer lasen Sätze außerhalb des Kernspintomographen und im Scanner leise oder laut mit entweder neutraler oder fröhlicher Intonation vor.
Die Verabreichung von Oxytocin erhöhte den zweiten Formanten der produzierten Vokale. Höhere Frequenzen dieses akustischen Parameters wurden zuvor mit einer positiven Valenz gesprochener Sprache in Verbindung gebracht; jedoch konnten unabhängige Beurteiler*innen die akustischen Unterschiede in unserem experimentellen Setting nicht konsistent unterscheiden.
Als neuronales Korrelat verstärkte Oxytocin die präparatorische subkortikale Gehirnaktivität im ventralen Pallidum und Striatum. Auch kortikal erhöhte Oxytocin präparatorische Gehirnaktivität in Regionen des dorsalen wie auch des ventralen Sprachverarbeitungsstroms, in sensomotorischen Kortizes und limbischen sowie exekutiven Regionen. In einigen dieser Regionen modulierte der genetische Oxytocin- Rezeptor-Polymorphismus rs53576 die durch die Oxytocin-Verabreichung verursachte Gehirnaktivität. Ähnlich wie Dopamin modulierte Oxytocin außerdem kortiko-basale Schaltkreise, die an der Generierung von fröhlicher Prosodie beteiligt sind. Während der Vorbereitung von Sprache erhöhte der Neuromodulator die funktionelle Konnektivität zwischen dem ventralem Pallidum und dem dorsolateralen präfrontalen Kortex mit einem spiegelbildlichen Profil während des eigentlichen Sprechens, einen Effekt den wir als „gating“ (Bahnung) interpretierten.
Unsere Ergebnisse legen nahe, dass mehrere neuronale Prozesse, die der Sprachproduktion zugrundeliegen, durch Oxytocin moduliert werden. Das Muster ähnelt hierbei dem anderer Neuromodulatoren wie Dopamin. Die vorliegende Arbeit charakterisiert somit erstmals Oxytocineffekte auf die mit Sprachproduktion assoziierte Hirnaktivität und funktionelle Konnektivität.
Bergjan, Manuela ; Tannen, Antje ; Mai, Tobias ; Feuchtinger, Johanna ; Luboeinski, Jennifer ; Bauer, Julian ; Fischer, Uli ; Kocks, Andreas
Einleitung : Eine sinnvolle Einbindung von Pflegefachpersonen mit Hochschulabschluss in die Versorgungsabläufe wird international häufig mit besseren Behandlungsergebnissen bei den Patient*innen assoziiert. In Deutschland fehlt es derzeit noch an verlässlichen Zahlen über Absolvent*innen und deren Aufgabenfeldern. Ziel dieser Erhebung war daher, durch Wiederholung einer früheren Erhebung erneut den Anteil von Pflegefachpersonen mit Bachelor- oder Masterabschlüssen in der direkten Patient*innenversorgung zu ermitteln.
Methode: In einer Querschnittserhebung wurden die Pflegedirektor*innen der Universitätskliniken und Medizinischen Hochschulen (UK) Deutschlands mittels einer standardisierten Befragung nach der Anzahl der Pflegefachpersonen mit Hochschulabschlüssen (Bachelor, Master und Doktor) gefragt. Weitere Fragen betrafen deren Aufgabengebiete und Integrationsmaßnahmen. Die Daten wurden mittels deskriptiver Statistik ausgewertet.
Ergebnisse: Insgesamt konnten n = 29 gültige Fragebögen aus 35 UK in die Analyse eingeschlossen werden, daraus ergibt sich eine Rücklaufquote von 82,85%. Für insgesamt 18 UK konnte eine Steigerung der hochschulisch qualifizierten Pflegefachpersonen um n = 786, von 2015 (n = 593) auf 2018 (n = 1379) erreicht werden. Der Anteil an Pflegefachpersonen mit Hochschulabschluss in den teilnehmenden UK liegt bei 3,16% (SD = 1,66; Min - Max = 1,09 - 6,69; Q1 - Q3 = 1,49 - 4,04; 95% KI = 2,30 – 3,95). In der direkten Versorgung beträgt der Anteil 2,11% (SD = 1,40; Min – Max = 0,47 - 5,42; Q1 – Q3 = 0,87 – 3,16; 95% KI 1,36 - 2,76). Die Aufgabenschwerpunkte liegen im Bereich der Regelversorgung und Patient*innenedukation (Bachelorabsolvent*innen), der evidenzbasierten Pflegepraxisentwicklung (Masterabsolvent*innen) und Forschung (promovierte Absolvent*innen).
Diskussion: Im Vergleich zu 2015 ist der Anteil hochschulisch qualifizierter Pflegefachpersonen zwar angestiegen, doch er liegt immer noch auf einem sehr niedrigen Niveau. Die Hochschulabsolvent*innen nehmen versorgungs- und entwicklungsrelevante Aufgaben wahr, doch besteht hinsichtlich ihrer Aufgabengebiete Bedarf an kompetenzorientierter Differenzierung.
Klinker, Hartwig ; Naumann, Uwe ; Rössle, Martin ; Berg, Thomas ; Bondin, Mark ; Lohmann, Kristina ; Koenig, Bettina ; Zeuzem, Stefan ; Cornberg, Markus
Glecaprevir/pibrentasvir, a pangenotypic, direct-acting antiviral combination approved for chronic hepatitis C virus treatment, has limited real-world evidence supporting 8-week therapy in compensated cirrhosis. We investigated effectiveness and safety of 187 hepatitis C virus-infected, treatment-naïve, patients with compensated cirrhosis receiving 8-week glecaprevir/pibrentasvir therapy in the German Hepatitis C-Registry between 2 August 2017 and 1 January 2020. Sustained virologic response was 98.4% (127/129) in the per-protocol analysis (excluding patients lost to follow-up or who discontinued treatment due to compliance) and was 85.8% (127/148) in patients with data available in an intention-to-treat analysis. Nineteen patients were lost to follow-up; nine genotype 3 patients, nine nongenotype 3 patients and one mixed genotype patient. One patient relapsed, and one died, unrelated to treatment. Adverse events (>5%) were fatigue and headache. Two serious adverse events occurred; no adverse events resulted in drug discontinuation. An 8-week glecaprevir/pibrentasvir therapy was effective and well-tolerated in this real-world analysis.
Hintergrund: Bei der Operation einer ATAD sind Patienten aufgrund multipler komplexer Faktoren gefährdet perioperative permanente neurologische Defizite zu erleiden. Da perioperative PND die Mortalität signifikant steigern, ist die Kenntnis über potentielle Risikofaktoren für ein PND von großem Wert, nicht zuletzt um bestmöglich auf jeden Patientenfall vorbereitet sein zu können und Therapiestrategien zu optimieren.
Diese retrospektive Studie soll prä- und intraoperative Risikofaktoren für die Entstehung eines PND nach der Operation einer ATAD herausfiltern.
Material und Methoden: Patientendaten von Patienten mit ATAD (n=305), die sich im Zeitraum von 2001 – 2017 am Universitätsklinikum Frankfurt in der Abteilung für Herz- und Gefäßchirurgie einer Operation unterzogen haben, wurden retrospektiv mittels univariater Analyse und multivariater logistischer Regression analysiert.
Ergebnisse: Die PND-Rate innerhalb der Studienpopulation betrug 13%. Mit hoher statistischer Signifikanz konnte eine Form der hämodynamischen Instabilität als präoperativer Risikofaktor für die Entstehung eines perioperativen PND identifiziert werden (OR 9,53; p<0.001). Weiterhin konnte gezeigt werden, dass das Vorhandensein einer Karotisstenose das perioperative PND-Risiko ungünstig beeinflusst (OR 2,68, p=0,04). Ein präoperativer Sinusrhythmus kann die perioperative PND-Rate günstig beeinflussen (OR 0,2, p=0,01). Die univariate Analyse konnte signifikant belegen, dass Operationszeiten > 300 Minuten und EKZ-Zeiten > 160 Minuten das PND-Risiko ungünstig beeinflussen. Andere Risikofaktoren wie z.B. die Art der Hirnperfusion oder der Grad des hypothermischen Kreislausstillstandes, die zumindest klinische Signifikanz zu haben scheinen, konnten in dieser Arbeit keine statistische Signifikanz erzielen, was ggf. Ausdruck der Limitationen retrospektiver Arbeiten ist.
Fazit: Eine hämodynamische Instabilität stellt einen präoperativen Risikofaktor für die Entstehung eines PND nach der Operation einer ATAD dar. Zu den identifizierten präoperativen Risikofaktoren, die die PND-Rate ungünstig beeinflussen gehört außerdem das Vorhandensein einer Karotisstenose, während das Vorhandensein eines Sinusrhythmus die PND-Rate günstig beeinflusst.
Das Zeitmanagement bei der Operation einer ATAD ist entscheidend, um peri-operativen PND vorbeugen zu können. Eine Operationszeit > 300 Minuten und eine EKZ-Zeit von > 160 Minuten sind mit wesentlich höheren PND-Raten assoziiert und stellen somit intraoperative Risikofaktorenfür die Entstehung eines PND bei der Operation einer ATAD dar.
Die akute Nierenschädigung ist ein häufiges klinisches Erscheinungsbild, das trotz der heutigen Erkenntnisse über pathophysiologische Abläufe in der Niere mit einer erhöhten Morbidität und Mortalität assoziiert ist. Die eigene Fähigkeit der Niere zur Regeneration stellt ein Potenzial dar, das durch die Unterstützung pro-regenerativer Faktoren das Patientenüberleben verbessern kann. Das Wissen, dass die akute Nierenschädigung ein reversibles Ereignis darstellt, bestärkt den Einsatz der Forschung pro-regenerative Einflussfaktoren zu bestimmen, deren Zusammenhang darzustellen und eine mögliche Strategie zur innovativen Therapie zu entwickeln. Um eine akute Nierenschädigung darzustellen und anschließend auf regenerative Prozesse zu untersuchen, wurde ein Cisplatin-induziertes in vitro-Schädigungsmodell an primären Tubulusepithelzellen (mTEZ) aus Wildtyp Mäusen etabliert. Nach Isolation und Kultivierung primärer mTEZ erfolgte die Schädigung mit Cisplatin, die anhand eines Zytotoxizitätsnachweises quantifiziert wurde. Makrophagen zeichnen sich durch ihre funktionale Vielfalt in physiologischen als auch pathophysiologischen Abläufen aus. Ihre Plastizität ermöglicht es ihnen, sich entsprechend des umgebenden Milieus mit ihrem Phänotyp anzupassen und folglich in Form eines pro-regenerativen Makrophagen Proliferation und Reparaturprozesse zu unterstützen. Für die Untersuchung einer Makrophagen-vermittelten, pro-regenerativen Wirkung auf geschädigte mTEZ wurden primäre Zellen aus dem Knochenmark von Mäusen isoliert und zu Makrophagen differenziert. Zur Ausprägung eines pro-regenerativen Makrophagen Phänotyps erfolgte die Stimulation der kultivierten Makrophagen durch Inkubation mit Interleukin-10 (IL-10) und die Herstellung eines konditionierten Mediums (KM). Lipocalin-2 (Lcn-2) ist bekannt als früher Biomarker im Rahmen der akuten Nierenschädigung, aber zeichnet sich zusätzlich durch seine pro-proliferative Wirkung und regenerative Funktion aus. Lcn-2 ist ein Protein, das Eisen mit hoher Affinität bindet und in Makrophagen als alternativer Eisen-Transportmechanismus dient. In der vorliegenden Untersuchung stellte sich bei Stimulation mit IL-10 ein pro-regenerativer Makrophagen Phänotyp dar, der sich durch eine erhöhte Eisenfreisetzung und dem erhöhten Nachweis von Eisen-beladenen Lcn-2 im KM auszeichnete (holo-Lcn-2). Um den Zusammenhang von Lcn-2 aus IL-10-stimulierten Makrophagen und die regenerativen Eigenschaften auf mTEZ zu untersuchen, wurde ein Versuchsaufbau etabliert, indem mTEZ mit Cisplatin geschädigt und anschließend ein KM von IL-10-stimulierten Wildtyp (WT) oder Lcn-2 knockout Makrophagen hinzugefügt wurde. Zusätzlich wurde ein rekombinantes holo-Lcn-2 hergestellt, das als Zugabe zu KM von Lcn-2 knockout Makrophagen der Wiederherstellung und der Untersuchung eines Lcn-2-abhängigen Mechanismus diente. Als Merkmal einer Zellregeneration wurden die epitheliale Integrität und die Reorganisation des Zytoskeletts bestimmt. Ergänzend konnte mit Hilfe der Expression von Proliferationsmarkern sowie einer Echtzeitmessung der Proliferationsrate eine zunehmende Proliferation geschädigter mTEZ nach Zugabe von KM aus Makrophagen in Abhängigkeit von Lcn-2 bewiesen werden. Anschließend wurde eine Analyse des Eisengehalts im Zelllysat von mTEZ durchgeführt. Hierbei konnte ein signifikanter Anstieg des Eisengehaltes in mTEZ nach Zugabe von KM aus WT Makrophagen als auch durch Ergänzung von rekombinanten holo-Lcn-2 zu KM aus Lcn-2 knockout Makrophagen nachgewiesen werden. In der Korrelation zwischen Eisenmenge im Zelllysat der mTEZ und der Proliferationsrate ergab sich eine zunehmende Proliferation mit Anstieg des Eisengehaltes der Zelle. Zusammenfassend ergaben unsere Untersuchungen, dass KM aus pro-regenerativen Makrophagen die Überlebensfähigkeit von mTEZ nach Cisplatin-Schädigung steigert. Es zeigte sich auch eine durch Lcn-2 geförderte epitheliale Integrität sowie ein pro-proliferativer Effekt. Die regenerativen Effekte an mTEZ wurden durch Lcn-2 aus KM von IL-10-stimulierten Makrophagen über seine Eisen-bindende Funktion vermittelt. Über die Ausschüttung von Lcn-2 vermitteln pro-regenerative Makrophagen vermutlich die Zell-Regeneration von mTEZ, indem Lcn-2 toxisches Eisen von geschädigten und apoptotischen Zellen aus der Umgebung bindet, es Zielzellen als holo-Lcn-2 zur Verfügung stellt und hierdurch die Proliferation induziert.
Schmidt, Florian ; Marx, Alexander ; Baumgarten, Nina ; Hebel, Marie ; Wegner, Martin ; Kaulich, Manuel ; Leisegang, Matthias ; Brandes, Ralf ; Göke, Jonathan ; Vreeken, Jilles ; Schulz, Marcel Holger
Understanding how epigenetic variation in non-coding regions is involved in distal gene-expression regulation is an important problem. Regulatory regions can be associated to genes using large-scale datasets of epigenetic and expression data. However, for regions of complex epigenomic signals and enhancers that regulate many genes, it is difficult to understand these associations. We present StitchIt, an approach to dissect epigenetic variation in a gene-specific manner for the detection of regulatory elements (REMs) without relying on peak calls in individual samples. StitchIt segments epigenetic signal tracks over many samples to generate the location and the target genes of a REM simultaneously. We show that this approach leads to a more accurate and refined REM detection compared to standard methods even on heterogeneous datasets, which are challenging to model. Also, StitchIt REMs are highly enriched in experimentally determined chromatin interactions and expression quantitative trait loci. We validated several newly predicted REMs using CRISPR-Cas9 experiments, thereby demonstrating the reliability of StitchIt. StitchIt is able to dissect regulation in superenhancers and predicts thousands of putative REMs that go unnoticed using peak-based approaches suggesting that a large part of the regulome might be uncharted water.
Highlights
• Single nucleotide variants (SNVs) may affect transcription factor (TF) binding
• Fast statistical approach to assess significance of differential TF binding for SNVs
• Validate new approach on in vitro and in vivo TF binding assays
• Applications on GWAS SNVs and large eQTL studies illustrate utility
Summary
Non-coding variants located within regulatory elements may alter gene expression by modifying transcription factor (TF) binding sites, thereby leading to functional consequences. Different TF models are being used to assess the effect of DNA sequence variants, such as single nucleotide variants (SNVs). Often existing methods are slow and do not assess statistical significance of results. We investigated the distribution of absolute maximal differential TF binding scores for general computational models that affect TF binding. We find that a modified Laplace distribution can adequately approximate the empirical distributions. A benchmark on in vitro and in vivo datasets showed that our approach improves upon an existing method in terms of performance and speed. Applications on eQTLs and on a genome-wide association study illustrate the usefulness of our statistics by highlighting cell type-specific regulators and target genes. An implementation of our approach is freely available on GitHub and as bioconda package.
Osteocalcin (auch Bone 6 la Protein [B G P] genannt), ein 49 Aminosäuren enthaltendes Knochenmatrixprotein, wird in den Osteoblasten synthetisiert und bei gesteigertem Knochenumsatz im Serum erhöht gefunden. Osteocalcin im Serum kann mittels eines Hadioimmunoassays gemessen werden. Die Werte steigen mit dem Lebensalter an und zeigen einen zirkadianen Rhythmus. Beim primären Hyperparathyreoidismus sind die Osteocalcinwerte erhöht. Bei Patienten mit Mammacarcinom zeigt ein Osteocalcinanstieg stets eine Metastasierung im Knochen an. Die Hyperthyreose geht mit erhöhten Osteocalcinwerten einher. Über den diagnostischen Wert einer Osteocalcinbestimmung beim Plasmozytom läßt sich zur Zeit noch keine sichere Aussage machen. Patienten mit einer histomorphometrisch gesicherten „low-turnover'"Osteoporose weisen niedrige Osteocalclnspiegel auf. Die stark erhöhten Osteocalcinwerte bei der renalen Osteopathie (sekundärer Hyperparathyreoidismus, Osteomalazie) sind z.T. auch au feine verminderte renale Elimination von Osteocalcin bei eingeschränkter glomerulärer Filtrationsrate zurückzuführen. Zusammenfassend stellt die Osteocalcinbestimmung eine Bereicherung der diagnostischen Möglichkeiten zur Beurteilung des Knochenumsatzes darf sollte vorerst jedoch noch vorwiegend wissenschaftlichen Fragestellungen vorbehalten sein.
Im Zeitraum von 1991 - 1992 wurden 2000 Stuhlproben von a) Tropenreisenden (n = 600), b) Patienten mit Durch fall (n = 500) und c) Patienten mit gastrointestinalen Symptomen, jedoch ohne Durchfall (n = 900) auf Campylobacter spp. untersucht Hierfür wurden drei Kultivierungsmethoden parallel eingesetzt: 7. eine Filtermembranmethode, 2. ein blutfreies Campylobacter-Selektivmedium und 3. ein Blutmedium mit Butzler-Supplement Zusätzlich wurde der Einfluß von Transportbedingungen untersucht mittels Vergleiches zwischen Nativ-Stuhlproben versus Stühlen, die in ein Transportnährmedium (Cary-Blair-Medium) gegeben worden waren (n = 517).
Insgesamt waren 54/2000 (2,7%) Stuhlproben positiv. Die Campylobacter-Auffindungsrate in flüssigen Stühlen (Gruppe B) lag dreifach höher im Vergleich zu nicht-durchfälligen Proben (p < 0.01). In den Gruppen b) und c) waren Patienten, die älter als 40 Jahre waren, signifikant seltener mit Campylobacter infiziert als jüngere Personen, während es bei den Tropenreisenden keinen Alterseffekt gab. Unter den drei Kultivierungsmethoden war das Filtermembranverfahren dasjenige mit der besten Campylobacter-Ausbeute (p < 0.01): Methode 1 ergab 57,5%, Methode 2 50% und Methode 3 40,7% positive Isolate. Durch den Einsatz von Cary-Blair-Transportmedium konnte die Campylobacter-Isolierungsrate um 25% gesteigert werden. Die am häufigsten isolierten Spezies waren C. jejuni und C. coli.
Das neue immunoluminometrische BeriLux® PSA - ein Test für das Routinelabor. Ein Methodenvergleich
(1993)
In dieser Studie wurde der immunoluminometrische BeriLux® PSA-Test mit zwei radio-immunologischen und einem fluorometrischen Verfahren verglichen. Für den BeriLux® PSA wurden an 150 gesunden Probanden (100 Männer und 50 Frauen) die Referenzbereiche ermittelt. Bei Männern lag die 95%-Perzentile bei 3,77 ng/ml, für Frauen lag die 95%-Perzentile bei0,1 ng/ml. Der Korrelationskoeffizient zwischen der immunoluminometrischen Methode und radioimmunologischen Methode liegt bei r = 0,99, die analytische Sensitivität von BeriLux9 PSA liegt bei 0,03 ng/ml. Die Stabilität der Serumproben bei Lagerungstemperaturen zwischen 2°C und 8°C ist über einen Tag garantiert. Über diesen Zeitraum hinaussollten Serumproben bei -20° C gelagert werden.
Koch, Vitali ; Loos, Gerald ; Grünewald, Leon David ; Eichler, Katrin ; Booz, Christian ; D’Angelo, Tommaso ; Yel, Ibrahim ; Mahmoudi, Scherwin ; Martin, Simon S. ; Harth, Marc ; Albrecht, Moritz Hans Ernst ; Zangos, Stephan ; Bernatz, Simon ; Thalhammer, Axel ; Scholtz, Jan-Erik ; Vogl, Thomas J. ; Gruber-Rouh, Tatjana
Purpose: To assess the diagnostic precision of three different workstations for measuring thoracic aortic aneurysms (TAAs) in vivo and ex vivo using either pre-interventional computed tomography angiography scans (CTA) or a specifically designed phantom model.
Methods: This retrospective study included 23 patients with confirmed TAA on routinely performed CTAs. In addition to phantom tube diameters, one experienced blinded radiologist evaluated the dimensions of TAAs on three different workstations in two separate rounds. Precision was assessed by calculating measurement errors. In addition, correlation analysis was performed using Pearson correlation.
Results: Measurements acquired at the Siemens workstation deviated by 3.54% (range, 2.78–4.03%; p = 0.14) from the true size, those at General Electric by 4.05% (range, 1.46–7.09%; p < 0.0001), and at TeraRecon by 4.86% (range, 3.22–6.45%; p < 0.0001). Accordingly, Siemens provided the most precise workstation at simultaneously most fluctuating values (scattering of 4.46%). TeraRecon had the smallest fluctuation (scattering of 2.83%), but the largest deviation from the true size of the phantom. The workstation from General Electric showed a scattering of 2.94%. The highest overall correlation between the 1st and 2nd rounds was observed with measurements from Siemens (r = 0.898), followed by TeraRecon (r = 0.799), and General Electric (r = 0.703). Repetition of measurements reduced processing times by 40% when using General Electric, by 20% with Siemens, and by 18% with TeraRecon.
Conclusions: In conclusion, all three workstations facilitated precise assessment of dimensions in the majority of cases at simultaneously high reproducibility, ensuring accurate pre-interventional planning of thoracic endovascular aortic repair.
Background: Opsoclonus-myoclonus syndrome (OMS) is a rare, immune-mediated neurological disorder. In adults, the pathogenesis can be idiopathic, post-infectious or paraneoplastic, the latter etiology belonging to the ever-expanding group of defined paraneoplastic neurological syndromes (PNS). In contrast to other phenotypes of PNS, OMS cannot be ascribed to a single pathogenic autoantibody. Here, we report the first detailed case of paraneoplastic, antibody-negative OMS occurring in association with a pancreatic neuroendocrine tumor (pNET).
Case presentation: A 33-year-old female presented with a two-week history of severe ataxia of stance and gait, dysarthria, head tremor, myoclonus of the extremities and opsoclonus. Her past medical history was notable for a metastatic pancreatic neuroendocrine tumor, and she was subsequently diagnosed with paraneoplastic opsoclonus-myoclonus syndrome. Further workup did not reveal a paraneoplastic autoantibody. She responded well to plasmapheresis, as she was refractory to the first-line therapy with corticosteroids.
Conclusions: This case expands current knowledge on tumors associated with paraneoplastic opsoclonus-myoclonus syndrome and the age group in which it can occur. It further adds evidence to the effectiveness of plasmapheresis in severe cases of opsoclonus-myoclonus syndrome with a lack of response to first-line therapy.
Dinh, Truc Sophia ; Brückle, Maria-Sophie ; González-González, Ana I. ; Witte, Julian ; Akker, Marjan van den ; Gerlach, Ferdinand M. ; Muth, Christiane
Structured management programs have been developed for single diseases but rarely for patients with multiple medications. We conducted a qualitative study to investigate the views of stakeholders on the development and implementation of a polypharmacy management program in Germany. Overall, we interviewed ten experts in the fields of health policy and clinical practice. Using content analysis, we identified inclusion criteria for the selection of suitable patients, the individual elements that should make up such a program, healthcare providers and stakeholders that should be involved, and factors that may support or hinder the program’s implementation. All stakeholders were well aware of polypharmacy-related risks and challenges, as well as the urgent need for change. Intervention strategies should address all levels of care and include all concerned patients, caregivers, healthcare providers and stakeholders, and involved parties should agree on a joint approach.
Zielsetzung: Ziel der Studie ist die Frage, ob es möglich ist, einen Kommunikationskurs für ca. 400 Studierende erfolgreich in ein Blended Learning-Format (asynchrone Theorievermittlung/digitaler synchroner Praxisteil) umzustellen. Hauptfokus war dabei die Beurteilung des subjektiven Lernzuwachses und die Frage, inwiefern die Bedeutung des Themas Kommunikation und Gesprächsführung online abgebildet werden kann. Die Betrachtung erfolgt auf Basis von Evaluationsergebnissen der Studierenden und Dozierenden dieses Kurses.
Methodik: Die Studierenden des vierten vorklinischen Semesters aus dem Jahr 2020 wurden durch einen Selbsteinschätzungsbogen zu Beginn sowie durch eine Evaluation nach Abschluss des Kurses befragt. Zusätzlich wurden die Rückmeldungen der Dozierenden zum Kurs ausgewertet. Um die Ergebnisse vergleichen und mögliche Unterschiede feststellen zu können, wurden die entsprechenden Selbsteinschätzungs- und Evaluationsergebnisse der vorangegangenen zehn Jahre (in Form von Präsenzkursen) herangezogen.
Ergebnisse: Im Onlineformat wurde ein deutlicher subjektiver Lernzuwachs angegeben, der höher ausfiel als in den Vorjahren in Präsenz. Die Eignung des Onlineformats wurde von Studierenden sowie Dozierenden eher kritisch bewertet, während die Kursatmosphäre positiv gesehen wurde. Die Relevanz ärztlicher Gesprächsführung wurde im Onlineformat insgesamt besonders gut bewertet.
Schlussfolgerung: Aufgrund der Evaluationsergebnisse werden die Erfahrungen mit dem Blended Learning-Format auch in Zukunft in das Kommunikations-Curriculum des Medizinstudiums der Goethe-Universität Frankfurt einfließen. Die Ergebnisse haben gezeigt, dass das Erlernen ärztlicher Gesprächsführung online gut möglich ist. Daher kann dieses Format für zukünftige neue Konzepte empfohlen werden.
Mahmoudi, Scherwin ; Lange, Marvin ; Lenga, Lukas ; Yel, Ibrahim ; Koch, Vitali ; Booz, Christian ; Martin, Simon ; Bernatz, Simon ; Vogl, Thomas J. ; Albrecht, Moritz Hans Ernst ; Scholtz, Jan-Erik
Objectives: To assess the impact of noise-optimised virtual monoenergetic imaging (VMI+) on image quality and diagnostic evaluation in abdominal dual-energy CT scans with impaired portal-venous contrast.
Methods: We screened 11,746 patients who underwent portal-venous abdominal dual-energy CT for cancer staging between 08/2014 and 11/2019 and identified those with poor portal-venous contrast.
Standard linearly-blended image series and VMI+ image series at 40, 50, and 60 keV were reconstructed. Signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) of abdominal organs and vascular structures were calculated. Image noise, image contrast and overall image quality were rated by three radiologists using 5-point Likert scale.
Results: 452 of 11,746 (4%) exams were poorly opacified. We excluded 190 cases due to incomplete datasets or multiple exams of the same patient with a final study group of 262. Highest CNR values in all abdominal organs (liver, 6.4 ± 3.0; kidney, 17.4 ± 7.5; spleen, 8.0 ± 3.5) and vascular structures (aorta, 16.0 ± 7.3; intrahepatic vein, 11.3 ± 4.7; portal vein, 15.5 ± 6.7) were measured at 40 keV VMI+ with significantly superior values compared to all other series. In subjective analysis, highest image contrast was seen at 40 keV VMI+ (4.8 ± 0.4), whereas overall image quality peaked at 50 keV VMI+ (4.2 ± 0.5) with significantly superior results compared to all other series (p < 0.001).
Conclusions: Image reconstruction using VMI+ algorithm at 50 keV significantly improves image contrast and image quality of originally poorly opacified abdominal CT scans and reduces the number of non-diagnostic scans.
Advances in knowledge: We validated the impact of VMI+ reconstructions in poorly attenuated DECT studies of the abdomen in a big data cohort.
März, Winfried ; Baumstark, Manfred W. ; Scharnagl, Hubert ; Pohl, Tilla ; Ruzicka, Viktor ; Herwig, Jürgen ; Schaaf, Ludwig ; Ruß, Andreas ; Böhles, Hans-Josef ; Usadel, Klaus Henning ; Groß, Werner
Measurements of vertical velocity from vertically pointing Doppler lidars are used to derive the profiles of vertical velocity variance. Observations were taken during the FESSTVaL (Field Experiment on Submesoscale Spatio-Temporal Variability in Lindenberg) campaign during the warm seasons of 2020 and 2021. Normalized by the square of convective velocity scale, the average vertical velocity variance profile follows the universal profile of Lenschow et al. (1980), however, daily profiles still show a high day-to-day variability. We found that moisture transport and the content of moisture in the boundary layer could explain the remaining variability of the normalized vertical velocity variance. The magnitude of the normalized vertical velocity variance is highest on clear-sky days, and decreases as the relative humidity increase and surface latent heat flux decrease in cloud-topped and rainy days. This suggests that moisture content and moisture transport are limiting factors for the intensity of turbulence in the convective boundary layer. We also found that the intensity of turbulence decreases with an increase in boundary layer cloud fraction during FESSTVaL, while the latent heating in the cloud layer was not a relevant source of turbulence in this case. We conclude that a new vertical velocity scale has to be defined that would take into account the moist processes in the convective boundary layer.
Measurements of vertical velocity from vertically pointing Doppler lidars are used to derive the profiles of normalized vertical velocity variance. Observations were taken during the FESSTVaL (Field Experiment on Submesoscale Spatio-Temporal Variability in Lindenberg) campaign during the warm seasons of 2020 and 2021. Normalized by the square of the convective velocity scale, the average vertical velocity variance profile follows the universal profile of Lenschow et al. (1980). However, daily profiles still show a high day-to-day variability. We found that moisture transport and the content of moisture in the boundary layer could explain the remaining variability of the normalized vertical velocity variance. The magnitude of the normalized vertical velocity variance is highest on clear-sky days and decreases as the absolute humidity increases and surface latent heat flux decreases on cloud-topped days. This suggests that moisture content and moisture transport are limiting factors for the intensity of turbulence in the convective boundary layer. We also found that the intensity of turbulence decreases with an increase in the boundary layer cloud fraction during FESSTVaL, while the latent heating in the cloud layer was not a relevant source of turbulence in this case. We conclude that a new vertical velocity scale has to be defined that would take into account the moist processes in the convective boundary layer.
Ziel der vorliegenden Untersuchung war es, die Analytik der Neurotransmitterbestimmung im Liquor zu etablieren und in das vorhandene Leistungsspektrum des Stoffwechsellabors zu integrieren. Die klinische Ausrichtung gilt sowohl der Detektion primärer Neurotransmitterdefekte als auch der Untersuchung der Ursachen und Auswirkungen sekundärer Neurotransmitterstörungen.
Die Messung der Biogenen Amine im Liquor mittels der Hochdruck-Flüssigkeits-Chromatographie in dem hier vorgestellten Verfahren konnte standardisiert und in das vorhandene Leistungsspektrum des Stoffwechsellabors des Zentrums für Kinderheilkunde und Jugendmedizin der J.W. Goethe-Universität, Frankfurt etabliert werden. Es ist nachgewiesenermaßen sensitiv, spezifisch und erlaubt eine schnelle und reproduzierbare Analyse bei minimaler Probenmenge und Probenaufarbeitung.
Meßtechnisch konnte durch den von unserer Arbeitsgruppe verwendeten ESA CoulArray® Detektor (Modell 5600) (im Gegensatz zu den bisher beschriebenen amperometrischen Detektoren) aufgrund seiner 16 einzelnen coulometrischen Sensoren eine enorme Verbesserung der Nachweisqualität der untersuchten Neurotransmittersubstanzen erreicht werden. Aufgrund verschiedener Potentialfelder ist eine spezifische Detektion über einen weiten Einsatzbereich aufgrund der elektrochemischen Unterschiede zu erzielen. Dies ermöglicht eine wesentlich größere Anzahl zu detektierender Substanzen eindeutig zu identifizieren und erlaubt damit nicht nur die eindeutige Zuordnung der zu messenden Endabbauprodukte HVA und 5HIAA, sondern auch des gesamten Spektrums der vorgeschalteten Metabolite. Somit ließ sich mit diesem aufgebauten Analysesystem die komplexe vollständige Abbildung des Stoffwechselwegs der zentralen biogenen Amine verwirklichen.
Die sehr sensible Messung der Parameter stellt große Anforderung an
Probengewinnung, Probenverarbeitung und Meßanalyse. Aufgrund der
Durchführung interner Qualitäskontrolluntersuchungen konnte der Nachweis einer sicheren reproduzierbaren Analytik erbracht werden. Insbesondere die Notwendigkeit der ausreichenden Unterdrückung der weiter ablaufenden chemischen Prozesse durch die unmittelbare Kühlung der Liquorproben auf eine Temperatur von -80°C wurde nachgewiesen. Zusätzlich konnte gezeigt werden, dass die Verwendung von handelsüblichen EDTA-Röhrchen einen ausreichenden antioxidativen Schutz bietet und wir somit eine anwenderfreundliche Probengewinnung empfehlen können. Die strikte Einhaltung des Protokolls zur Gewinnung und weiteren Verarbeitung der Liquorproben ist elementar und stellt insbesondere bei zur Zeit noch bestehenden uneinheitlichen altersabhängigen Referenzwerten die Grundlage zur diagnostischen Bewertung der gemessenen Werte dar. Eine Standardisierung der Referenzwerte auf internationaler Ebene ist unverzichtbar zur korrekten Interpretation der gemessenen Daten und sollte vorrangiges Streben aller beteiligten Laboratorien sein. Ein entsprechender Antrag auf ein erneutes, jährlich stattfindendes externes Qualitätskontrollprogramm (Ringversuch) wurde gestellt.
Mit der Bestimmung der Biogenen Amine im Liquor steht ein potentes Instrument zur Detektion pädiatrischer Neurotransmittererkrankungen zur Verfügung. Es gibt weltweit jedoch nur eine geringe Anzahl von Laboratorien mit diesem Analyseangebot. Somit trägt die Etablierung dieser Analytik im vorhandenen Leistungsspektrum des Stoffwechsellabors des Zentrums für Kinderheilkunde und Jugendmedizin der J.W. Goethe-Universität, Frankfurt dazu bei diese Lücke zu schließen. In der Mehrzahl sind Erkrankungen der zentralen Biogenen Amine ausschließlich nur über eine Messung der Metaboliten im Liquor diagnostizierbar, nur wenige angeborene Neurotransmitterdefekte fallen durch periphere Serumparameter, wie z.B. einer Hyperphenylalaninämie auf. Es ergibt sich der Verdacht, dass diese Erkrankungsformen unterdiagnostiziert sind und auch das Wissen um diese Erkrankungen nicht allgemein vorausgesetzt werden kann.
Mit den Neurotransmittervorstufen (z.B. L-Dopa oder 5OH-Tryptophan) steht betroffenen Patienten eine adäquate Therapieoption zur Verfügung, so dass die Aufdeckung eines spezifischen Defektes in der Synthese der Biogenen Amine vorrangiges Ziel sein müsste. Exemplarisch für die beeindruckende Effektivität der Therapie mittels L-Dopa und 5OH-Tryptophan sei auf den Erfahrungsbericht des von unserer Arbeitsgruppe betreuten Patienten mit 6-PTPS-Defekt verwiesen.
Zusätzlich konnte mit dieser Arbeit gezeigt werden, dass die wiederholenden Messungen dem Therapiemonitoring dienen und eine unverzichtbare Grundlage der individuellen Therapie betroffener Patienten darstellen. Sekundäre Veränderungen der Neurotransmission sind beschrieben, jedoch fehlen weitergehende Untersuchungen zur Einordnung ihrer klinischen Relevanz. Gerade diese große inhomogene Patientengruppe stellt eine zukünftige Herausforderung für weitere wissenschaftliche Anstrengungen in diesem Bereich dar. Wie anhand des Beispiels der Patienten mit mitochondrialer Enzephalopathie dargestellt, kann die vorhandene Analysemethode dazu beitragen das Wissen um die Pathophysiologie bekannter Erkrankungen zu erweitern. Da analog zu den primären Erkrankungen auch hier Therapieoptionen zur Verfügung stehen würden, ist es dringend notwendig die übliche Liquordiagnostik diesbezüglich auszudehnen. Aufgrund der auch im Rahmen dieser Arbeit erhobenen Daten kann gefolgert werden bei ungeklärten neuropädiatrischen Erkrankungen, besonders jenen, welche in der Neonatal- oder frühen Säuglingszeit beginnen und progredient verlaufen, und sich klinisch durch Dyskinesien, Krampfanfälle und Enzephalopathien äußern, eine Liquoruntersuchung auf Biogene Amine zu veranlassen.
Somit dient die erbrachte Arbeit zur Etablierung der Analytik der Neurotransmitterbestimmung im Liquor dem Erkenntnisgewinn neuer metabolischer Erkrankungen, erweitert das Grundlagenwissen bekannter Störungen und dient der Detektion seltener, jedoch oft schwerwiegender Erkrankungen.
In der vorliegenden Arbeit präsentieren wir die Daten von perkutanen Interventionen im Bereich der Beckenstrombahn, welche retrospektiv der Klassifikation der Transatlantic Society Consensus Gruppe (TASC) folgend als TASC C und D kategorisiert wurden. Für TASC D Läsionen wurde von der TASC Gruppe die chirurgische Therapie empfohlen. Für TASC C Läsionen konnte mangels aussagekräftiger Daten keine konkrete Empfehlung bezüglich einer chirurgischen oder interventionellen Behandlung ausgesprochen werden und man berief sich auf die nach wie vor weit verbreitete Anwendung gefäßchirurgischer operativer Therapie im Fall dieser Läsionen.
Ziel dieser Arbeit war es, herauszuarbeiten, ob bei Behandlung von Gefäßläsionen dieser beiden Klassifikationen mit perkutaner transluminaler Angioplastie akzeptable Erfolgs- und Offenheitsraten erreicht werden können und ob eine Empfehlung zu Gunsten der interventionellen Therapie ausgesprochen werden kann.
In einem Zeitraum von Januar 2000 bis Dezember 2002 wurden an unserem Institut 253 Patienten mit 294 Läsionen im Bereich der Beckenarterien interventionell behandelt. Unter ihnen befanden sich 89 Patienten mit 129 klassifizierten TASC C und D Läsionen, 110 Stenosen (65 in der AIC und 45 in der AIE) und 19 Okklusionen (12 in der AIC und 7 in der AIE). Die durchschnittliche Länge der Läsion betrug 49,2±31,2 mm. Zum Einsatz kamen 148 Stents. Der technische Erfolg der Interventionen betrug 96,9%. Komplikationen traten bei 5,6% der Patienten auf, wobei wir in keinem dieser Fälle eine periphere Embolie verzeichneten. Die Nachbehandlung erfolgte über einen Zeitraum von 12-38 Monate, durchschnittlich 25 Monate. Nachuntersuchungen umfassten eine klinische Untersuchung, die Messung des Tibial-Brachialen-Quotienten und eine Duplex-Sonographie am ersten Tag postinterventionell, nach 1, 3, 6 und 12 Monaten und anschließend einmal jährlich.
Aus den Nachuntersuchungen ergab sich gemessen an den Kriterien der AHA ein klinischer Erfolg bei 97,3% der TASC C Patienten und 88,5% der Patienten mit TASC D Läsionen. Im Verlauf der Nachbeobachtungszeit kam es zu 6 Restenosen und einer Reokklusion im behandelten Gefäßsegment, von denen 4 erfolgreich mit Hilfe einer zweiten Intervention therapiert werden konnten. Bei 3 Patienten bedurfte es operativer Eingriffe in Form zweier aortoiliakaler Bypässe und einer Unterschenkelamputation.
Es ergab sich somit nach durchschnittlich 25 Monaten eine primäre Offenheitsrate von 89,9% und eine sekundäre Offenheitsrate von 95,5%. Die von uns präsentierten Daten sind mit den nach Behandlung fokaler, kurzstreckiger (<10 cm) Stenosen publizierten Daten vergleichbar.
Die Ergebnisse dieser Studie zeigen, dass die perkutane Rekanalisation sowohl im Falle der TASC C als auch der TASC D Läsionen mit hohem technischen und hohem klinischen Erfolg, einer niedrigen Komplikationsrate und viel versprechenden Offenheitsraten durchgeführt werden kann. Interventionelle Therapie kann somit durchaus auch für die Behandlung von Läsionen der Klassifikation TASC C und D empfohlen werden.
Bei einer fortgeschrittenen oro-vestibulären Alveolarfortsatzatrophie, die durch einen länger zurückliegenden Zahnverlust, Zahntrauma oder Entzündung bedingt ist, ist die Indikation zu einer Implantation häufig ohne zusätzliche Aufbaumaßnahmen des Alveolarkammes nicht möglich. Osteoplastische Verfahren, die der transversalen Kieferkammerweiterung dienen, können in diesen anatomisch schwierigen Ausgangsbedingungen zur Anwendung kommen.
Die vorliegende Studie sollte die Ergebnisse nach Anwendung eines minimal invasiven chirurgischen Verfahrens (Bone-spreading) mit Hilfe eines speziellen Instrumentensystems bewerten.
Über einen Zeitraum von 8 Jahren wurden bei 53 Patienten (24 Männern, 29 Frauen), die ein primär unzureichendes transversales Knochenangebot aufwiesen, 129 Implantate simultan mit der Bone-spreading-Technik inseriert. Die mittlere Liegedauer der Implantate betrug 3,81 Jahre. Zum Zeitpunkt der letzten Nachuntersuchung standen die Implantate durchschnittlich 3,39 Jahre unter funktioneller Belastung. Die Verteilung der inserierten Implantate erstreckte sich sowohl auf den Oberkiefer (57,36 % der Implantate) als auch den Unterkiefer (42,64 % der Implantate). Die prothetischen Versorgungen der 129 Implantate wurden mit 71 Implantaten als Krone, mit 44 als Brücke und mit jeweils 7 Implantaten als Konus- und Resilienzteleskope zementiert.
Die klinischen Daten (modifizierter Plaque-Index, modifizierter Sulcus-Blutungs-Index, Sondierungstiefe mesial und vestibulär, periimplantäre Mucosa, Breite der Gingiva propria, klinische Mobilität, Periotestwert) und die radiologischen Befunde (horizontaler und vertikaler Knochenabbau) wurden im Rahmen von jährlichen Recalluntersuchungen erhoben und analysiert.
Die Werte der Entzündungs-Indizes waren im allgemeinem stabil. So konnte bei 88 % der Patienten bei der letzten Nachuntersuchung kein höherer Wert von Plaqueindex als 1 (= nicht sichtbar, hauchdünner Belag) beobachtet werden. Auch der Sulcus-Blutungs-Index war zufriedenstellend. Hier wiesen 91 % der Patienten bei der letzten Nachuntersuchung keine Blutung bzw. nur einen Blutungspunkt, aber keine Farbveränderung oder Schwellung der Gingiva auf. Ähnlich verhielten sich die Werte der Sondierungstiefen. Diese Ergebnisse
spiegelten sich in den stabilen periimplantären Verhältnisse wider, die bei 93,81 % der Probanden bei der letzten Nachuntersuchung vorgefunden wurden. Die gemessenen Periotestwerte entsprachen denen eines osseointegrierten Implantates. Sie lagen bei der letzten Nachuntersuchung im Schnitt bei –2,7. Der radiologisch ermittelte periimplantäre Knochenabbau war bei den hier inserierten Implantaten nicht größer als 0,2 mm pro Jahr. So konnte bei 85,27 % der unter Funktion stehenden Implantate kein bzw. nur ein minimaler horizontaler Knochenabbau vorgefunden werden, bei weiteren 13,95 % ging der horizontale Knochenabbau nicht über ein Viertel der Implantatlänge hinaus. Ähnlich verhielt sich der vertikale Knochenabbau. Kein bzw. nur ein minimaler Knochenabbau war bei 59,06 % der Patienten, ein bis zu einem Viertel bei 39,09 % vorzufinden. Ein Implantatverlust ist während der gesamten Beobachtungszeit nicht eingetreten. Das entsprach einer Überlebensrate von 100%.
Die bisherige Untersuchung über 8 Jahre zeigte, daß die orale Rehabilitation mit osseointegrierten Implantaten unter Anwendung der Bone-spreading-Methode sowohl bei einem transversalen Knochenangebot ab einer Alveolarfortsatzbreite von 3 mm als auch in den Knochenklassen D1 und D2, die vornehmlich im Unterkiefer vorzufinden sind, erfolgreich ist.
Background: In the Computer Tomography imaging, examinations for the diagnosis of lesions of the upper abdomen currently use water-soluble, iodinated, non-ionic contrast agents with low molecular weight. One possibility to reduce the time of the examination and X-ray exposure is to increase the injection rate. However, higher injections rates lead to increased hypersensitivity reactions and extravasation rates. Furthermore, cardiac pump function does not always allow for the transportation of such a large volume within one heartbeat. With a contrast agent of higher iodine concentration, the injection rate may be reduced without decreasing the iodine delivery rate while reducing the volume load of the heart.
Aim: to compare the performance and image quality of two injection protocols of contrast medium for multiphasic CT imaging of malignant hepatic lesions; one using Imeron 300 at an injection rate of 5 ml/ sec and the second using Imeron 400 at an injection rate of 3,7 ml/ sec, for multiphasic CT imaging of malignant hepatic lesions, in order to optimise the iodine concentration and injection rate of the contrast agent Imeron in the Multislice Spiral-CT of the upper abdomen.
Materials and methods: the current prospective, single centre, double-blinded, randomised and interindividual comparison study included 50 patients (29 males and 21 females) with a mean age of 63,3 years. Patients were randomised to one of the two injection protocols. Image evaluation included qualitative assessment (technical quality, presence of artefacts and overall contrast quality) and quantitative assessment (measuring the difference in HU between the lesion and the surrounding hepatic tissue). The difference between both protocols was tested for statistical significance using the Wilcoxon-Mann-Whitney test and the Two-Sample t-test.
Results: there was no statistically significant difference between both protocols regarding the technical quality of images, both in the AP (p = 0,46) and in the venous phase (p = 0,48). Additionally, no statistically significant difference was found regarding the presence of artefacts related to the contrast medium, both in the AP (p = 0,46) and in the venous phase (p = 0,46), as well as regarding the overall contrast quality of images both in the AP (p = 0,50) and in the venous phase (p = 0,48). Quantitative assessment showed no statistically significant difference regarding the difference in HU measurement between the hepatic lesion and the surrounding hepatic tissue, both in the AP (p = 0,36) and in the venous phase (p = 0,92).
Conclusion: in the multiphasic CT imaging of the liver, reducing the injection rate of the contrast medium Imeron from 5 ml/ sec to 3,7 ml/ sec while increasing the iodine strength of the agent from 300 to 400 mg/ml, respectively, and thus keeping the iodine injection flow rate constant, produces similar signal intensities and results in similar technical, image and overall contrast qualities..
Keywords: Contrast-medium, injection rate, iodine concentration, hepatic malignancy, multiphasic CT
Die E3-Ubiquitinligase TRIM25 ist in verschiedenen humanen Tumoren verstärkt exprimiert, was häufig mit einer schlechten Prognose der betroffenen Patienten sowie dem Auftreten von Therapieresistenzen korreliert. Unsere Arbeitsgruppe konnte TRIM25 zuvor als Caspase-2 und -7 mRNA-bindendes Protein und negativen Regulator beider Caspasen in humanen Kolonkarzinomzellen identifizieren. Ein transienter TRIM25 Knockdown führt in Abhängigkeit der erhöhten Expression der jeweiligen Caspasen zur Sensitivierung der Kolonkarzinomzellen gegenüber Chemotherapeutika-induzierter Apoptose. Ein Ziel dieser Arbeit war, die Übertragbarkeit dieser Erkenntnisse auf einen loss-of-function-Ansatz mit stabilen TRIM25 Knockdown Zellen zu überprüfen. Die stabilen Knockdown Zellen sollten der späteren Etablierung eines Xenograftmodells dienen. Da zahlreiche TRIM Proteine bekannterweise eine vielseitige Rolle bei der Regulation sowohl onkogener als auch tumorsuppressiver Prozesse einnehmen, wurde als weitere maßgebliche Fragestellung dieser Arbeit der Einfluss von TRIM25 auf wichtige tumorigene Eigenschaften wie Proliferation, Migration, Zellzyklus und Inflammation untersucht. TRIM25-abhängige Effekte auf das Migrationsver halten von RKO-Zellen wurden im in vitro-Wundheilungsassay mit stabilen TRIM25 Knock down Zellen analysiert. Aufgrund hoher interexperimenteller Unterschiede im Migrations verhalten derselben Zellklone konnte hinsichtlich einer TRIM25-abhängigen Regulation der Migration jedoch keine eindeutige Aussage getroffen werden. Dagegen belegten die gezeigten Proliferationsassays eine signifikant vermehrte Proliferation von RKO-Zellen nach stabilem TRIM25 Knockdown. Dies legt eine tumorsuppressive Rolle von TRIM25 nahe. Durch flusszytometrische Analysen stabiler TRIM25 Knockdown und Kontrollzellen zeigten hinge gen keinen konsistenten Einfluss von TRIM25 auf den Zellzyklus von RKO-Zellen. Eine Sensitivierung von Kolonkarzinomzellen gegenüber Chemotherapeutika-induzierter Apoptose konnte auch in stabilen TRIM25 Knockdown Zellen nachgewiesen werden, während die für transiente Ansätze bekannte, TRIM25 Knockdown-abhängige Hochregulation von Caspase 2 und -7 dagegen deutlich geringer ausgeprägt war. Dies lässt vermuten, dass die Tumorzellen einer Hochregulation Zelltod-induzierender Proteine bei einem konstitutiven TRIM25Knockdown gegenregulieren. Aufgrund der aber nach wie vor nachweisbaren Sensitivierung der TRIM25 Knockdown Zellen gegenüber Apoptose, können zusätzliche, bisher noch nicht bekannte Mechanismen postuliert werden, welche zur Sensitivierung dieser Zellen gegenüber Apoptose beitragen. Die daraus abgeleitete, TRIM25-abhängige Apoptosehemmung spricht für einen „Überlebensmechanismus“, welcher maßgeblich zur Chemotherapieresis tenz von Kolonkarzinomzellen beitragen kann. Hinsichtlich eines Einflusses von TRIM25 auf entzündliche Prozesse wurden RKO-Zellen mit klassischen Aktivatoren des NLRP3-Inflammasoms stimuliert und ausgewählte Marker mittels Western Blot-Analysen nachgewiesen. TRIM25 Knockdown-abhängig war eine verminderte Spaltung der Apoptosemarker Caspase-3, -7 und PARP-1 nachweisbar. Caspase-7 und PARP-1 spielen bekanntermaßen auch im Rahmen Inflammasom-induzierter Signalprozesse eine wichtige Rolle, während die Spaltung von Caspase-3 durch das NLRP3-Inflammasom induziert werden kann und v.a. für apoptotische oder pyroptotische Prozesse verantwortlich gemacht wird. Daher kann postuliert werden, dass der stabile TRIM25 Knockdown zur Hemmung von Inflammasom-induzierten Signalprozessen führt und darüber Kolonkarzinomzellen vor Apoptose/Pyroptose geschützt werden. Umgekehrt deutet dies auf eine Aktivierung von Inflammasom-vermittelten Signalprozessen durch TRIM25 hin.
Aufgrund der hohen Inzidenz und Mortalität, der schlechten Prognose und der Entwicklung von Therapieresistenzen besteht ein dringender Bedarf in der Entwicklung neuer, verbesserter Therapien des kolorektalen Karzinoms sowie der Resensibilisierung der Krebszellen gegenüber gängigen, bereits bestehenden Therapieoptionen. Im Rahmen dieser Arbeit konnte ein onkogener Einfluss von TRIM25 auf Kolonkarzinomzellen durch Hemmung Chemotherapeutika-induzierter Apoptose nachgewiesen werden, während eine mögliche tumorigene Rolle von TRIM25 durch zusätzliche Aktivierung Inflammasom-induzierter Signalprozesse weiterer Untersuchungen bedarf. Zusammenfassend kann TRIM25 als vielversprechender therapeutischer Angriffspunkt für die Entwicklung von sowohl neuen antientzündlichen als auch tumorsuppressiven Therapien betrachtet werden.