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Rössig, Lothar ; Haendeler, Judith ; Hermann, Corinna ; Malchow, Philipp ; Urbich, Carmen ; Zeiher, Andreas M. ; Dimmeler, Stefanie
MAP kinase-dependent phosphorylation processes have been shown to interfere with the degradation of the antiapoptotic protein Bcl-2. The cytosolic MAP kinase phosphatase MAP kinase phosphatase-3 (MKP-3) induces apoptosis of endothelial cells in response to tumor necrosis factor alpha (TNFalpha) via dephosphorylation of the MAP kinase ERK1/2, leading to Bcl-2 proteolysis. Here we report that the endothelial cell survival factor nitric oxide (NO) down-regulated MKP-3 by destabilization of MKP-3 mRNA. This effect of NO was paralleled by a decrease in MKP-3 protein levels. Moreover, ERK1/2 was found to be protected against TNFalpha-induced dephosphorylation by coincubation of endothelial cells with the NO donor. Subsequently, both the decrease in Bcl-2 protein levels and the mitochondrial release of cytochrome c in response to TNFalpha were largely prevented by exogenous NO. In cells overexpressing MKP-3, no differences in phosphatase activity in the presence or absence of NO were found, excluding potential posttranslational modifications of MKP-3 protein by NO. These data demonstrate that upstream of the S-nitrosylation of caspase-3, NO exerts additional antiapoptotic effects in endothelial cells, which rely on the down-regulation of MKP-3 mRNA.
Ceramide levels are strongly increased by stimulation of renal mesangial cells with nitric oxide (NO). This effect was shown previously to be due to a dual action of NO, comprising an activation of sphingomyelinases and an inhibition of ceramidase activity. In this study we show that the NO-triggered inhibition of neutral ceramidase activity is paralleled by a down-regulation at the protein level. A complete loss of neutral ceramidase protein is obtained after 24 h of stimulation. Whereas the selective proteasome inhibitor lactacystin blocked NO-evoked ceramidase degradation, several caspase inhibitors were ineffective. Moreover, the NO-induced degradation is reversed by the protein kinase C (PKC) activator, 12-O-tetradecanoylphorbol-13-acetate (TPA), and also by the physiological PKC activators platelet-derived growth factor-BB (PDGF), angiotensin II and ATP, resulting in a normalization of neutral ceramidase protein as well as activity. In vivophosphorylation studies using 32Pi-labeled mesangial cells revealed that TPA, PDGF, angiotensin II, and ATP trigger an increased phosphorylation of the neutral ceramidase, which is blocked by the broad spectrum PKC inhibitor Ro-31 8220 but not by CGP 41251, which has a preferential action on Ca2+-dependent isoforms, thus suggesting the involvement of a Ca2+-independent PKC isoform. In vitro phosphorylation assays using recombinant PKC isoenzymes and neutral ceramidase immunoprecipitated from unstimulated mesangial cells show that particularly the PKC-δ isoform and to a lesser extent the PKC-α isoform are efficient in directly phosphorylating neutral ceramidase. In summary, our data show that NO is able to induce degradation of neutral ceramidase, thereby promoting accumulation of ceramide in the cell. This effect is reversed by PKC activation, most probably by the PKC-δ isoenzyme, which can directly phosphorylate and thereby prevent neutral ceramidase degradation. These novel regulatory interactions will provide therapeutically valuable information to target neutral ceramidase stability and subsequent ceramide accumulation.
Akool, El-Sayed ; Doller, Anke ; Müller, Roswitha ; Gutwein, Paul ; Xin, Cuiyan ; Huwiler, Andrea ; Pfeilschifter, Josef ; Eberhardt, Wolfgang
Excessive accumulation of the extracellular matrix is a hallmark of many inflammatory and fibrotic diseases, including those of the kidney. This study addresses the question whether NO, in addition to inhibiting the expression of MMP-9, a prominent metalloprotease expressed by mesangial cells, additionally modulates expression of its endogenous inhibitor TIMP-1. We demonstrate that exogenous NO has no modulatory effect on the extracellular TIMP-1 content but strongly amplifies the early increase in cytokine-induced TIMP-1 mRNA and protein levels. We examined whether transforming growth factor beta (TGFbeta), a potent profibrotic cytokine, is involved in the regulation of NO-dependent TIMP-1 expression. Experiments utilizing a pan-specific neutralizing TGFbeta antibody demonstrate that the NO-induced amplification of TIMP-1 is mediated by extracellular TGFbeta. Mechanistically, NO causes a rapid increase in Smad-2 phosphorylation, which is abrogated by the addition of neutralizing TGFbeta antisera. Similarly, the NO-dependent increase in Smad-2 phosphorylation is prevented in the presence of an inhibitor of TGFbeta-RI kinase, indicating that the NO-dependent activation of Smad-2 occurs via the TGFbeta-type I receptor. Furthermore, activation of the Smad signaling cascade by NO is corroborated by the NO-dependent increase in nuclear Smad-4 level and is paralleled by increased DNA binding of Smad-2/3 containing complexes to a TIMP-1-specific Smad-binding element (SBE). Reporter gene assays revealed that NO activates a 0.6-kb TIMP-1 gene promoter fragment as well as a TGFbeta-inducible and SBE-driven control promoter. Chromatin immunoprecipitation analysis also demonstrated DNA binding activity of Smad-3 and Smad-4 proteins to the TIMP-1-specific SBE. Finally, by enzyme-linked immunosorbent assay, we demonstrated that NO causes a rapid increase in TGFbeta(1) levels in cell supernatants. Together, these experiments demonstrate that NO by induction of the Smad signaling pathway modulates TIMP-1 expression.
Rössig, Lothar ; Fichtlscherer, Birgit ; Breitschopf, Kristin ; Haendeler, Judith ; Zeiher, Andreas M. ; Mülsch, Alexander ; Dimmeler, Stefanie
In cultured human endothelial cells, physiological levels of NO prevent apoptosis and interfere with the activation of the caspase cascade. In vitro data have demonstrated that NO inhibits the activity of caspase-3 by S-nitrosation of the enzyme. Here we present evidence for the in vivo occurrence and functional relevance of this novel antiapoptotic mechanism. To demonstrate that the cysteine residue Cys-163 of caspase-3 is S-nitrosated, cells were transfected with the Myc-tagged p17 subunit of caspase-3. After incubation of the transfected cells with different NO donors, Myc-tagged p17 was immunoprecipitated with anti-Myc antibody. S-Nitrosothiol was detected in the immunoprecipitate by electron spin resonance spectroscopy after liberation and spin trapping of NO by N-methyl-D-glucamine-dithiocarbamate-iron complex. Transfection of cells with a p17 mutant, where the essential Cys-163 was mutated into alanine, completely prevented S-nitrosation of the enzyme. As a functional correlate, in human umbilical vein endothelial cells the NO donors sodium nitroprusside or PAPA NONOate (50 microM) significantly reduced the increase in caspase-3-like activity induced by overexpressing caspase-3 by 75 and 70%, respectively. When human umbilical vein endothelial cells were cotransfected with beta-galactosidase, morphological analysis of stained cells revealed that cell death induction by overexpression of caspase-3 was completely suppressed in the presence of sodium nitroprusside, PAPA NONOate, or S-nitroso-L-cysteine (50 microM). Thus, NO supplied by exogenous NO donors serves in vivo as an antiapoptotic regulator of caspase activity via S-nitrosation of the Cys-163 residue of caspase-3.
Grimm, Oliver ; Nägele, Magdalena Maria ; Küpper-Tetzel, Lea ; Greck, Moritz de ; Plichta, Michael M. ; Reif, Andreas
Rationale: Dysregulation of dopaminergic neurotransmission, specifically altered reward processing assessed via the reward anticipation in the MID task, plays a central role in the etiopathogenesis of neuropsychiatric disorders. Objectives: We hypothesized to find a difference in the activity level of the reward system (measured by the proxy reward anticipation) under drug administration versus placebo, in that amisulpride reduces, and L-DOPA enhances, its activity. Methods: We studied the influence of dopamine agonist L-DOPA and the antagonist amisulpride on the reward system using functional magnetic resonance imaging (fMRI) during a monetary incentive delay (MID) task in n = 45 healthy volunteers in a randomized, blinded, cross-over study. Results: The MID paradigm elicits strong activation in reward-dependent structures (such as ventral striatum, putamen, caudate, anterior insula) during reward anticipation. The placebo effect demonstrated the expected significant blood oxygen level–dependent activity in reward-dependent brain regions. Neither amisulpride nor L-DOPA led to significant changes in comparison with the placebo condition. This was true for whole-brain analysis as well as analysis of a pre-defined nucleus accumbens region-of-interest mask. Conclusion: The present results cast doubt on the sensitivity of reward anticipation contrast in the MID task for assessing dopamine-specific changes in healthy volunteers by pharmaco-fMRI. While our task was not well-suited for detailed analysis of the outcome phase, we provide reasonable arguments that the lack of effect in the anticipation phase is not due to an inefficient task but points to unexpected behavior of the reward system during pharmacological challenge. Group differences of reward anticipation should therefore not be seen as simple representatives of dopaminergic states.
Wenzel, Mike ; Würnschimmel, Christoph ; Chierigo, Francesco ; Tian, Zhe ; Shariat, Shahrokh F. ; Terrone, Carlo ; Saad, Fred ; Tilki, Derya ; Graefen, Markus ; Mandel, Philipp ; Kluth, Luis ; Chun, Felix ; Karakiewicz, Pierre I.
Background: To test for rates of other cause mortality (OCM) and cancer-specific mortality (CSM) in elderly prostate cancer (PCa) patients treated with the combination of radical prostatectomy (RP) and external beam radiation therapy (EBRT) versus RP alone, since elderly PCa patients may be over-treated. Methods: Within the Surveillance, Epidemiology and End Results database (2004–2016), cumulative incidence plots, after propensity score matching for cT-stage, cN-stage, prostate specific antigen, age and biopsy Gleason score, and multivariable competing risks regression models (socioeconomic status, pathological Gleason score) addressed OCM and CSM in patients (70–79, 70–74, and 75–79 years) treated with RP and EBRT versus RP alone. Results: Of 18,126 eligible patients aged 70–79 years, 2520 (13.9%) underwent RP and EBRT versus 15,606 (86.1%) RP alone. After propensity score matching, 10-year OCM rates were respectively 27.9 versus 20.3% for RP and EBRT versus RP alone (p < .001), which resulted in a multivariable HR of 1.4 (p < .001). Moreover, 10-year CSM rates were respectively 13.4 versus 5.5% for RP and EBRT versus RP alone. In subgroup analyses separately addressing 70–74 year old and 75–79 years old PCa patients, 10-year OCM rates were 22.8 versus 16.2% and 39.5 versus 24.0% for respectively RP and EBRT versus RP alone patients (all p < .001). Conclusion: Elderly patients treated with RP and EBRT exhibited worrisome rates of OCM. These higher than expected OCM rates question the need for combination therapy (RP and EBRT) in elderly PCa patients and indicate the need for better patient selection, when combination therapy is contemplated.
Koyanagi, Masamichi ; Haendeler, Judith ; Badorff, Cornel ; Brandes, Ralf ; Hoffmann, Jörg ; Pandur, Petra D. ; Zeiher, Andreas M. ; Kühl, Michael ; Dimmeler, Stefanie
Human endothelial circulating progenitor cells (CPCs) can differentiate to cardiomyogenic cells during co-culture with neonatal rat cardiomyocytes. Wnt proteins induce myogenic specification and cardiac myogenesis. Here, we elucidated the effect of Wnts on differentiation of CPCs to cardiomyogenic cells. CPCs from peripheral blood mononuclear cells were isolated from healthy volunteers and co-cultured with neonatal rat cardiomyocytes. 6–10 days after co-culture, cardiac differentiation was determined by α-sarcomeric actinin staining of human lymphocyte antigen-positive cells (fluorescence-activated cell-sorting analysis) and mRNA expression of human myosin heavy chain and atrial natriuretic peptide. Supplementation of co-cultures with Wnt11-conditioned medium significantly enhanced the differentiation of CPCs to cardiomyocytes (1.7 ± 0.3-fold), whereas Wnt3A-conditioned medium showed no effect. Cell fusion was not affected by Wnt11-conditioned medium. Because Wnts inhibit glycogen synthase kinase-3β, we further determined whether the glycogen synthase kinase-3β inhibitor LiCl also enhanced cardiac differentiation of CPCs. However, LiCl (10 mm) did not affect CPC differentiation. In contrast, Wnt11-conditioned medium time-dependently activated protein kinase C (PKC). Moreover, the PKC inhibitors bisindolylmaleimide I and III significantly blocked differentiation of CPCs to cardiomyocytes. PKC activation by phorbol 12-myristate 13-acetate significantly increased CPC differentiation to a similar extent as compared with Wnt11-conditioned medium. Our data demonstrate that Wnt11, but not Wnt3A, augments cardiomyogenic differentiation of human CPCs. Wnt11 promotes cardiac differentiation via the non-canonical PKC-dependent signaling pathway.
The adult heart has a limited capacity to replace or regenerate damaged cardiac tissue following severe myocardial injury. Thus, therapies facilitating the induction of cardiac regeneration holds great promise for the treatment of end-stage heart failure, and for pathologies invoking severe cardiac dysfunction as a result of cardiomyocyte death. Recently, a number of studies have demonstrated that cardiac regeneration can be achieved through modulation and/or reprogramming of cardiomyocyte proliferation, differentiation, and survival signaling. Non-coding RNAs (ncRNAs), including microRNAs (miRNAs), long non-coding RNAs (lncRNAs) and circular RNAs (circRNAs), are reported to play critical roles in regulating key aspects of cardiomyocyte physiologic and pathologic signaling, including the regulation of cardiac regeneration both in vitro and in vivo. In this review, we will explore and detail the current understanding of ncRNA function in cardiac regeneration, and highlight established and novel strategies for the treatment of heart failure through modulation of ncRNAs-driven cardiac regeneration.
Knop, Viola ; Hoppe, Daniel ; Vermehren, Johannes ; Trötschler, Sven ; Herrmann, Eva ; Vermehren, Annika ; Friedrich-Rust, Mireen ; Sarrazin, Christoph ; Trebicka, Jonel ; Zeuzem, Stefan ; Welker, Martin-Walter
Long-term effects on cirrhosis and portal hypertension of direct antiviral agent (DAA)-based eradication of hepatitis C virus (HCV) are still under debate. We analysed dynamics of liver and spleen elastography to assess potential regression of cirrhosis and portal hypertension 3 years post-treatment. Fifty-four patients with HCV-associated cirrhosis and DAA-induced SVR were included. Liver and spleen stiffness were measured at baseline (BL), end of treatment (EOT), 24 weeks after EOT (FU24) and 1, 2 and 3 (FU144) years post-treatment by transient liver elastography (L-TE) and point shear wave elastography (pSWE) using acoustic radiation force impulse (ARFI) of the liver (L-ARFI) and spleen (S-ARFI). Biochemical, virological and clinical data were also obtained. Liver stiffness assessed by L-TE decreased between BL [median (range), 32.5(9.1–75) kPa] and EOT [21.3(6.7–73.5) kPa; p < .0001] and EOT and FU144 [16(4.1–75) kPa; p = .006]. L-ARFI values improved between EOT [2.5(1.2–4.1) m/s] and FU144 [1.7(0.9–4.1) m/s; p = .001], while spleen stiffness remained unchanged. Overall, L-TE improved in 38 of 54 (70.4%) patients at EOT and 29 of 38 (76.3%) declined further until FU144, whereas L-ARFI values decreased in 30/54 (55.6%) patients at EOT and continued to decrease in 28/30 (93.3%) patients at FU144. Low bilirubin and high albumin levels at BL were associated with improved L-ARFI values (p = .048) at EOT or regression of cirrhosis (<12.5 kPa) by L-TE at FU144 (p = .005), respectively. Liver stiffness, but not spleen stiffness, continued to decline in a considerable proportion of patients with advanced liver disease after HCV eradication.
Faragli, Alessandro ; Alogna, Alessio ; Lee, Chong Bin ; Zhu, Miry ; Ghorbani, Niky ; Lo Muzio, Francesco Paolo ; Schnackenburg, Bernhard ; Stehning, Christian ; Kühne, Titus ; Post, Heiner ; Goubergrits, Leonid ; Nagel, Eike ; Pieske, Burkert ; Kelle, Sebastian ; Kelm, Marcus
Background: Myocardial efficiency should be maintained stable under light-to-moderate stress conditions, but ischemia puts the myocardium at risk for impaired functionality. Additionally, the measurement of such efficiency typically requires invasive heart catheterization and exposure to ionizing radiation. In this work, we aimed to non-invasively assess myocardial power and the resulting efficiency during pharmacological stress testing and ischemia induction. Methods: In a cohort of n = 10 healthy Landrace pigs, dobutamine stress testing was performed, followed by verapamil-induced ischemia alongside cardiac magnetic resonance (CMR) imaging. External myocardial power, internal myocardial power, and myocardial efficiency were assessed non-invasively using geometrical and functional parameters from CMR volumetric as well as blood flow and pressure measurements. Results: External myocardial power significantly increased under dobutamine stress [2.3 (1.6–3.1) W/m2 vs. 1.3 (1.1–1.6) W/m2, p = 0.005] and significantly decreased under verapamil-induced ischemia [0.8 (0.5–0.9) W/m2, p = 0.005]. Internal myocardial power [baseline: 5.9 (4.6–8.5) W/m2] was not affected by dobutamine [7.5 (6.9–9.0) W/m2, p = 0.241] nor verapamil [5.8 (4.7–8.8) W/m2, p = 0.878]. Myocardial efficiency did not change from baseline to dobutamine [21% (15–27) vs. 31% (20–44), p = 0.059] but decreased significantly during verapamil-induced ischemia [10% (8–13), p = 0.005]. Conclusion: In healthy Landrace pigs, dobutamine stress increased external myocardial power, whereas myocardial efficiency was maintained stable. On the contrary, verapamil-induced ischemia substantially decreased external myocardial power and myocardial efficiency. Non-invasive CMR was able to quantify these efficiency losses and might be useful for future clinical studies evaluating the effects of therapeutic interventions on myocardial energetics.
Given the simplicity of the method and how it can be applied, as well as proof that it lowers the mortality rate, fecal occult blood testing (FOBT) is currently the most commonly used screening method for colorectal cancer (CRC). However, the test suffers from poor sensitivity, particularly with respect to detecting early stages, as well as low acceptance among the population. Preliminary data on detecting calprotectin and tumour-M2-PK in the stool indicated that a better screening performance could be expected. But these tests also suffer from low sensitivity in detecting early stages and from poor specificity, thus limiting the usefulness of the tests as a result of high follow-up costs. Recently developed immunological tests (I-FOBT) demonstrate significantly increased sensitivity and specificity. I-FOBTs use antibodies specific to human hemoglobin and are therefore not affected by diet and drugs, leading to improved patient partipication. At present, I-FOBTs seem to be the most cost-effective approach for non-invasive screening. The detection of tumour-DNA in the stool opens up a new era in early diagnosis of colorectal cancer. Small trials have pointed to a very high sensitivity of these methods: 62–91% for colorectal cancer and between 26% and 73% for adenomas, with a very high level of specificity (93–100%). The major drawback of this type of testing, compared with other screening tests available today, is its high cost.
Non-organ confined stage and upgrading rates in exclusive PSA high-risk prostate cancer patients
(2022)
Höh, Robert Benedikt ; Flammia, Rocco Simone ; Hohenhorst, Lukas ; Sorce, Gabriele ; Chierigo, Francesco ; Tian, Zhe ; Saad, Fred ; Gallucci, Michele ; Briganti, Alberto ; Terrone, Carlo ; Shariat, Shahrokh F. ; Graefen, Markus ; Tilki, Derya ; Kluth, Luis ; Mandel, Philipp ; Becker, Andreas ; Chun, Felix ; Karakiewicz, Pierre I.
Background: The pathological stage of prostate cancer with high-risk prostate-specific antigen (PSA) levels, but otherwise favorable and/or intermediate risk characteristics (clinical T-stage, Gleason Grade group at biopsy [B-GGG]) is unknown. We hypothesized that a considerable proportion of such patients will exhibit clinically meaningful GGG upgrading or non-organ confined (NOC) stage at radical prostatectomy (RP).
Materials and methods: Within the Surveillance, Epidemiology, and End Results database (2010–2015) we identified RP-patients with cT1c-stage and B-GGG1, B-GGG2, or B-GGG3 and PSA 20–50 ng/ml. Rates of GGG4 or GGG5 and/or rates of NOC stage (≥ pT3 and/or pN1) were analyzed. Subsequently, separate univariable and multivariable logistic regression models tested for predictors of NOC stage and upgrading at RP.
Results: Of 486 assessable patients, 134 (28%) exhibited B-GGG1, 209 (43%) B-GGG2, and 143 (29%) B-GGG3, respectively. The overall upgrading and NOC rates were 11% and 51% for a combined rate of upgrading and/or NOC stage of 53%. In multivariable logistic regression models predicting upgrading, only B-GGG3 was an independent predictor (odds ratio [OR]: 5.29; 95% confidence interval [CI]: 2.21–14.19; p < 0.001). Conversely, 33%–66% (OR: 2.36; 95% CI: 1.42–3.95; p = 0.001) and >66% of positive biopsy cores (OR: 4.85; 95% CI: 2.84–8.42; p < 0.001), as well as B-GGG2 and B-GGG3 were independent predictors for NOC stage (all p ≤ 0.001).
Conclusions: In cT1c-stage patients with high-risk PSA baseline, but low- to intermediate risk B-GGG, the rate of upgrading to GGG4 or GGG5 is low (11%). However, NOC stage is found in the majority (51%) and can be independently predicted with percentage of positive cores at biopsy and B-GGG.
Krüger, Caroline ; Schäfer, Ingmar ; Bussche, Hendrik van den ; Baehr, Michael ; Bickel, Horst ; Fuchs, Angela ; Gensichen, Jochen ; Maier, Wolfgang ; Riedel-Heller, Steffi Gerlinde ; König, Hans-Helmut ; Dahlhaus, Anne Maren ; Schön, Gerhard ; Weyerer, Siegfried ; Wiese, Birgitt ; Renteln-Kruse, Wolfgang von ; Langebrake, Claudia ; Scherer, Martin
Background: The elderly population deals with multimorbidity (three chronic conditions) and increasinged drug use with age. A comprehensive characterisation of the medication – including prescription and over-the-counter (OTC) drugs – of elderly patients in primary care is still insufficient.
Objectives: This study aims to characterise the medication (prescription and OTC) of multimorbid elderly patients in primary care and living at home by identifying drug patterns to evaluate the relationship between drugs and drug groups and reveal associations with recently published multimorbidity clusters of the same cohort.
Methods: MultiCare was a multicentre, prospective, observational cohort study of 3189 multimorbid patients aged 65 to 85 years in primary care in Germany. Patients and general practitioners were interviewed between 2008 and 2009. Drug patterns were identified using exploratory factor analysis. The relations between the drug patterns with the three multimorbidity clusters were analysed with Spearman-Rank-Correlation.
Results: Patients (59.3% female) used in mean 7.7 drugs; in total 24,535 drugs (23.7% OTC) were detected. Five drug patterns for men (drugs for obstructive pulmonary diseases (D-OPD), drugs for coronary heart diseases and hypertension (D-CHD), drugs for osteoporosis (D-Osteo), drugs for heart failure and drugs for pain) and four drug patterns for women (D-Osteo, D-CHD, D-OPD and drugs for diuretics and gout) were detected. Significant associations between multimorbidity clusters and drug patterns were detectable (D-CHD and CMD: male: ρ = 0.376, CI 0.322–0.430; female: ρ = 0.301, CI 0.624–0.340).
Conclusion: The drug patterns demonstrate non-random relations in drug use in multimorbid elderly patients and systematic associations between drug patterns and multimorbidity clusters were found in primary care.
The small intestine is a part of the gastrointestinal tract in which digestion and absorption of nutrients takes place. The small bowel follows the stomach and is followed by the large intestine, reaching from the pylorus to the valve of Bauhin and is separated into the duodenum, the jejunum, and the ileum.
Capsule endoscopy (CE) has the potential to offer a perfect overview of the small-bowel mucosa and complete visualization of the entire small bowel is achieved in most cases. In this video, there is an overview offered on normal findings in small-bowel CE and typical anatomical landmarks are indicated. This article is part of an expert video encyclopedia.
Basic, Michael ; Kubesch, Alica ; Kuhnhenn, Lisa ; Görgülü, Esra ; Finkelmeier, Fabian ; Dietz, Julia ; Knabe, Mate ; Mücke, Victoria Therese ; Mücke, Marcus Maximilian ; Berger, Annemarie ; Zeuzem, Stefan ; Sarrazin, Christoph ; Hildt, Eberhard ; Peiffer, Kai-Henrik
Background & Aims: HBV genotype G (HBV/G) is mainly found in co-infections with other HBV genotypes and was identified as an independent risk factor for liver fibrosis. This study aimed to analyse the prevalence of HBV/G co-infections in healthy European HBV carriers and to characterize the crosstalk of HBV/G with other genotypes.
Methods: A total of 560 European HBV carriers were tested via HBV/G-specific PCR for HBV/G co-infections. Quasispecies distribution was analysed via deep sequencing, and the clinical phenotype was characterized regarding qHBsAg-/HBV-DNA levels and frequent mutations. Replicative capacity and expression of HBsAg/core was studied in hepatoma cells co-expressing HBV/G with either HBV/A, HBV/D or HBV/E using bicistronic vectors.
Results: Although no HBV/G co-infection was found by routine genotyping PCR, HBV/G was detected by specific PCR in 4%-8% of patients infected with either HBV/A or HBV/E but only infrequently in other genotypes. In contrast to HBV/E, HBV/G was found as the quasispecies major variant in co-infections with HBV/A. No differences in the clinical phenotype were observed for HBV/G co-infections. In vitro RNA and DNA levels were comparable among all genotypes, but expression and release of HBsAg was reduced in co-expression of HBV/G with HBV/E. In co-expression with HBV/A and HBV/E expression of HBV/G-specific core was enhanced while core expression from the corresponding genotype was markedly diminished.
Conclusions: HBV/G co-infections are common in European inactive carriers with HBV/A and HBV/E infection, but sufficient detection depends strongly on the assay. HBV/G regulated core expression might play a critical role for survival of HBV/G in co-infections.
Wiechmann, Svenja ; Maisonneuve, Pierre ; Grebbin, Britta Moyo ; Hoffmeister, Meike ; Kaulich, Manuel ; Clevers, Hans ; Rajalingam, Krishnaraj ; Kurinov, Igor ; Farin, Henner ; Sicheri, Frank ; Ernst, Andreas
The small GTPases H, K, and NRAS are molecular switches that are indispensable for proper regulation of cellular proliferation and growth. Mutations in this family of proteins are associated with cancer and result in aberrant activation of signaling processes caused by a deregulated recruitment of downstream effector proteins. In this study, we engineered novel variants of the Ras-binding domain (RBD) of the kinase CRAF. These variants bound with high affinity to the effector binding site of active Ras. Structural characterization showed how the newly identified mutations cooperate to enhance affinity to the effector binding site compared to RBDwt. The engineered RBD variants closely mimic the interaction mode of naturally occurring Ras effectors and as dominant negative affinity reagent block their activation. Experiments with cancer cells showed that expression of these RBD variants inhibits Ras signaling leading to a reduced growth and inductions of apoptosis. Using the optimized RBD variants, we stratified patient-derived colorectal cancer organoids according to Ras dependency, which showed that the presence of Ras mutations was insufficient to predict sensitivity to Ras inhibition.
Schmidt, Michael ; Hoehl, Sebastian ; Berger, Annemarie ; Zeichhardt, Heinz ; Hourfar, Michael Kai ; Ciesek, Sandra ; Seifried, Erhard
Background: In the pandemic, testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by real-time polymerase chain reaction is one of the pillars on which countermeasures are based. Factors limiting the output of laboratories interfere with the effectiveness of public health measures. Conserving reagents by pooling samples in low-probability settings is proposed but may cause dilution and loss of sensitivity. Blood transfusion services had experience in performance of high throughput nucleic acid testing (NAT) analysis and can support the national health system by screening of the inhabitants for SARS-COV-2.
Methods: We evaluated a new approach of a multiple-swab method by simultaneously incubating multiple respiratory swabs in a single tube. Analytical sensitivity was constant up to a total number of 50 swabs. It was consequently applied in the testing of 50 symptomatic patients (5-sample pools) as well as 100 asymptomatic residents of a nursing home (10-sample pools).
Results: The novel method did not cause false-negative results with nonsignificantly differing cycle threshold values between single-swab and multiple-swab NAT. In two routine applications, all minipools containing positive patient samples were correctly identified.
Conclusions: The new method enables countries to increase the total number of testing significantly. The multiple-swab method is able to screen system relevant groups of employees frequently. The example in Germany shows that blood transfusion services can support general health systems with their experience in NAT and their high-throughput instruments. Screening of a huge number of inhabitants is currently the only option to prevent a second infection wave and enable exit strategies in many countries.
Vasella, Mauro Giovanni ; Wagner, Ulrich ; Fritz-Kuisle, Christine ; Seidl, Kati ; Giudici, Luca ; Exner, Gerhard Ulrich ; Moch, Holger ; Wild, Peter Johannes ; Bode-Lasniewska, Beata
BCOR-rearranged sarcomas are rare and belong to the Ewing-like sarcomas (ELS). Their morphology and histopathological features make the diagnosis challenging. We present a case, initially diagnosed as an unusual extraskeletal myxoid chondrosarcoma (EMC). A 54-year-old male patient developed an asymptomatic swelling of the lower leg. Imaging showed a 9.5-cm large intramuscular soft tissue mass. Due to its morphological and immunohistochemical profile on biopsy, it was initially diagnosed as an EMC. The patient was treated by complete resection and adjuvant radiotherapy and remained free of tumor at 7 years follow-up. Using next-generation sequencing (NGS), we retrospectively identified RGAG1-BCOR gene fusion (confirmed by RT-PCR), which has not been described in somatic soft tissue tumors so far. This finding broadens the spectrum of partner genes in the BCOR-rearranged sarcomas in a tumor with a well-documented, long clinical follow-up.
Chen, Jin-Ran ; Lazarenko, Oxana P. ; Blackburn, Michael L. ; Chen, Jennifer F. ; Randolph, Christopher E. ; Zabaleta, Jovanny ; Schröder, Katrin ; Pedersen, Kim B. ; Ronis, Martin
Tightly regulated and cell-specific NADPH-oxidases (Nox) represent one of the major sources of reactive oxygen species (ROS) signaling molecules that are involved in tissue development and stem cell self-renewal. We have characterized the role of Nox4 in osteo-progenitors during postnatal bone development. Nox4 expression in bone and ROS generation were increased during early osteoblast differentiation and bone development. Stromal osteoblastic cell self-renewal, proliferation and ROS production were significantly lower in samples from whole-body Nox4 knockout mice (Nox4-/-) and conditional knockout (CKO) mice with depletion of Nox4 in the limb bud mesenchyme compared with those from control mice (Nox4fl/fl), but they were reversed after 9 passages. In both sexes, bone volume, trabecular number and bone mineral density were significantly lower in 3-week old CKO and Nox4-/- mice compared with Nox4fl/fl controls. This was reflected in serum levels of bone formation markers alkaline phosphatase (ALP) and procollagen 1 intact N-terminal propeptide (P1NP). However, under-developed bone formation in 3-week old CKO and Nox4-/- mice quickly caught up to levels of control mice by 6-week of age, remained no different at 13-week of age, and was reversed in 32-week old male mice. Osteoclastogenesis showed no differences among groups, however, CTX1 reflecting osteoclast activity was significantly higher in 3-week old male CKO and Nox4-/- mice compared with control mice, and significantly lower in 32-week old Nox4-/- mice compared with control mice. These data suggest that Nox4 expression and ROS signaling in bone and osteoblastic cells coordinately play an important role in osteoblast differentiation, proliferation and maturation.
Nucleoredoxin (NXN) is a redox regulator of Disheveled and thereby of WNT signaling. Deficiency in mice leads to cranial dysmorphisms and defects of heart, brain, and bone, suggesting defects of cell fate determination. We used shRNA-mediated knockdown of NXN in SH-SY5Y neuroblastoma cells to study its impact on neuronal cells. We expected that shNXN cells would easily succumb to redox stress, but there were no differences in viability on stimulation with hydrogen peroxide. Instead, the proliferation of naïve shNXN cells was increased with a higher rate of mitotic cells in cell cycle analyses. In addition, basal respiratory rates were higher, whereas the relative change in oxygen consumption upon mitochondrial stressors was similar to control cells. shNXN cells had an increased expression of redox-sensitive heat shock proteins, Hsc70/HSPA8 and HSP90, and autophagy markers suggested an increase in autophagosome formation upon stimulation with bafilomycin and higher flux under low dose rapamycin. A high rate of self-renewal, autophagy, and upregulation of redox-sensitive chaperones appears to be an attractive anti-aging combination if it were to occur in neurons in vivo for which SH-SY5Y cells are a model.
Ein professioneller Orchestermusiker verbringt die meiste Zeit in körperlich ungünstiger Sitzhaltung beim Spielen. Die Folge ist ein Anstieg des Risikos für die Entwicklung von muskuloskelettalen Beschwerden [44, 127, 128]. Eine Verbesserung der Arbeitsbedingungen lässt sich u.a. durch den Einsatz von ergonomischen Stühlen erzielen, da sie einen Einfluss auf die Körperhaltung des Orchestermusikers besitzen. Im Mittelpunkt der vorliegenden Arbeit standen daher sechs unterschiedliche von der Firma Mey für Orchestermusiker konzipierte Stühle. Die Studie beinhaltete eine Untersuchung des Einflusses der Stühle auf die Oberkörperstatik und die Sitzdruckverteilung von Orchestermusikern und den Einfluss auf ihr Instrumentalspiel im Vergleich zur habituellen Sitzhaltung. Das Probandenkollektiv umfasste 24 Berufsmusiker des Polizeiorchesters Mainz (Rheinland-Pfalz, Deutschland) und bestand zum größten Teil aus Blasinstrumentalisten (3 Frauen, 21 Männer). Das Durchschnittsalter betrug 45 Jahre. Die Überprüfung der Oberkörperstatik erfolgte durch einen 3D-Rückenscanner (ABW GmbH, Frickenhausen, Deutschland), eine Evaluation der Druckverhältnisse im Gesäß durch eine Druckmessmatte (GeBioM GmbH, Münster, Deutschland), womit sich bei jedem Stuhl ein bestimmtes Druckmuster kennzeichnen ließ. Die Messung erfolgte pro Stuhl und Messgerät stets im Wechsel zwischen der statischen Position ohne Instrument (oI) und der statischen Position mit Instrument (mI). Bei der statistischen Auswertung kam es zur Verwendung nicht parametrischer Tests (Friedman-, Wilcoxon Matched- Pairs-Test), wobei das Signifikanzniveau bei ≤0,05 lag. Es erfolgte eine Unterteilung in einen Inter- und einen Intrastuhlvergleich.
Die Ergebnisse des Interstuhlvergleichs zeigten bezüglich der Schulterregion keine signifikanten Veränderungen, wohingegen im Hinblick auf die WS-Parameter Signifikanzen zwischen Stuhl 2 und 5 verzeichnet wurden: in der habituellen Position offenbarte die Rumpflänge D die größte Abweichung mit einem Längenunterschied von 14mm (p≤0,001), so auch die Rumpflänge S (16 mm; p≤0,001). Bezüglich dieser Stühle wiesen die restlichen WS-Parameter Abweichungen von max. 4° bzw. 3mm auf. Zwischen Stuhl 2 und 3 ergaben sich m.I. im Hinblick auf den thorakalen (p≤0,01) und lumbalen Biegungswinkel (p≤0,001) max. Diskrepanzen von 2,5°. Die größten Unterschiede in der Beckenregion zeigten sich beim Beckenabstand zwischen Stuhl 3 und 5 o.I. (7mm) und m.I. (4mm), (beide Bedingungen p≤0,001). Im Hinblick auf die Druckparameter fand sich eine Abhängigkeit zwischen belasteter Fläche und Sitzbeinhöckerdruck (SBH): eine kleine Fläche bedeutete eine schlechte Druckverteilung und umgekehrt. Stühle 1 und 4 besaßen den geringsten SBH (p≤0,001). Im Intrastuhlvergleich zeigten die Bereiche der WS, Schultern und Becken jeweils mindestens einen signifikanten Parameter auf, wie z.B. Schulterblattabstand, sagittale Rumpfneigung und Beckenabstand. Korrelationen zwischen den Parametern waren nicht zu erkennen. M.I. kommt es hinsichtlich des SBH auf der linken Hälfte im Schnitt auf allen Stühlen zu einer Druckerhöhung von 8,46%, auf der rechten zu einer von 11,11%. Im Hinblick auf den Oberschenkeldruck (OS) vollzieht sich die größte Veränderung (7,4bar) der rechten Gesäßhälfte auf Stuhl 2 mit p≤0,001. Der Interstuhlvergleich zeigt also, dass die Wahl eines Stuhls keine Auswirkung auf die Körperhaltung hat. Ursache für Diskrepanzen hinsichtlich des SBH ist die unterschiedliche Polsterung und Größe der Sitzfläche, welche eine hohe Relevanz in Bezug auf die Umverteilung des Drucks und den subjektiven Komfort besitzt. Eine gepolsterte und große Oberfläche ist gleichzusetzen mit einer günstigen Druckverteilung und einem angenehmen individuellen Sitzgefühl. Der Intrastuhlvergleich offenbart ebenfalls keine klinisch relevanten Veränderungen im Oberkörper. Ausschließlich in der Druckverteilung ist eine signifikante Variabilität hinsichtlich des OS rechts bei Stuhl 2 vorhanden (p≤0,001). Die Sitzposition der Probanden ist symmetrisch. Die Symmetrie bezieht sich sowohl auf den Schulter-, WS- und Beckenbereich, als auch auf die Druckverhältnisse im Gesäßbereich.
In der vorliegenden Studie konnte belegt werden, dass gepolsterte und breite Sitzoberflächen mit gleichmäßiger Druckverteilung und gleichzeitig hohem Komfort einhergehen. Im Hinblick auf das Musizieren über einen längeren Zeitraum ist das Vorhandensein eines hohen Komforts für den Orchestermusiker von Bedeutung. Diese Erkenntnisse sind bei der Weiterentwicklung von ergonomischen Stühlen zu berücksichtigen. Eine Analyse der Schulter- und Rumpfmuskulatur und Messung des Beckenwinkels ist in weiteren Studien zusätzlich erforderlich, um zu erforschen, inwieweit die Stühle die Fehlfunktionen des Bewegungsapparates beeinflussen.
The coronavirus disease 2019 COVID-19 pandemic is rapidly spreading worldwide and is becoming a major public health crisis. Increasing evidence demonstrates a strong correlation between obesity and the COVID-19 disease. We have summarized recent studies and addressed the impact of obesity on COVID-19 in terms of hospitalization, severity, mortality, and patient outcome. We discuss the potential molecular mechanisms whereby obesity contributes to the pathogenesis of COVID-19. In addition to obesity-related deregulated immune response, chronic inflammation, endothelium imbalance, metabolic dysfunction, and its associated comorbidities, dysfunctional mesenchymal stem cells/adipose-derived mesenchymal stem cells may also play crucial roles in fueling systemic inflammation contributing to the cytokine storm and promoting pulmonary fibrosis causing lung functional failure, characteristic of severe COVID-19. Moreover, obesity may also compromise motile cilia on airway epithelial cells and impair functioning of the mucociliary escalators, reducing the clearance of severe acute respiratory syndrome coronavirus (SARS-CoV-2). Obese diseased adipose tissues overexpress the receptors and proteases for the SARS-CoV-2 entry, implicating its possible roles as virus reservoir and accelerator reinforcing violent systemic inflammation and immune response. Finally, anti-inflammatory cytokines like anti-interleukin 6 and administration of mesenchymal stromal/stem cells may serve as potential immune modulatory therapies for supportively combating COVID-19. Obesity is conversely related to the development of COVID-19 through numerous molecular mechanisms and individuals with obesity belong to the COVID-19-susceptible population requiring more protective measures.
Attention selects relevant information regardless of whether it is physically present or internally stored in working memory. Perceptual research has shown that attentional selection of external information is better conceived as rhythmic prioritization than as stable allocation. Here we tested this principle using information processing of internal representations held in working memory. Participants memorized four spatial positions that formed the endpoints of two objects. One of the positions was cued for a delayed match-non-match test. When uncued positions were probed, participants responded faster to uncued positions located on the same object as the cued position than to those located on the other object, revealing object-based attention in working memory. Manipulating the interval between cue and probe at a high temporal resolution revealed that reaction times oscillated at a theta rhythm of 6 Hz. Moreover, oscillations showed an anti-phase relationship between memorized but uncued positions on the same versus other object as the cued position, suggesting that attentional prioritization fluctuated rhythmically in an object-based manner. Our results demonstrate the highly rhythmic nature of attentional selection in working memory. Moreover, the striking similarity between rhythmic attentional selection of mental representations and perceptual information suggests that attentional oscillations are a general mechanism of information processing in human cognition. These findings have important implications for current, attention-based models of working memory.
Recently, pertussis has become a problem also in the adult population, with incidences even higher than in children. Pediatric health care workers (HCWs) are an important source of transmission, exposing very young and immunocompromised patients to an increased risk of potentially severe pertussis infections. Encouraging HCWs to get vaccinated can play a vital role in stopping the transmission of pertussis, thereby reducing institutional outbreaks.
In Germany, HCWs come up with all sorts of reasons for not getting pertussis vaccination. This study was meant to provide information in order to better understand the backgrounds of these attitudes.
A survey was conducted at the children's university hospital in Frankfurt, using an anonymous questionnaire. Survey results were used to design an intervention to increase the immunization rate of staff. Disappointingly, our efforts to increase the acceptance of the immunization program by providing information in advance were not yet satisfying.
Misconception about pertussis vaccination was prevalent especially among nursing staff. The main reasons for non-compliance included: unawareness of an own risk of infection, the belief that pertussis is not a serious illness, fear of side effects, the belief that the pertussis vaccine might trigger the pertussis disease itself, and skepticism about the efficacy of the pertussis vaccination.
ecently, pertussis has become a problem also in the adult population, with incidences even higher than in children. Pediatric health care workers (HCWs) are an important source of transmission, exposing very young and immunocompromised patients to an increased risk of potentially severe pertussis infections. Encouraging HCWs to get vaccinated can play a vital role in stopping the transmission of pertussis, thereby reducing institutional outbreaks.
In Germany, HCWs come up with all sorts of reasons for not getting pertussis vaccination. This study was meant to provide information in order to better understand the backgrounds of these attitudes.
A survey was conducted at the children's university hospital in Frankfurt, using an anonymous questionnaire. Survey results were used to design an intervention to increase the immunization rate of staff. Disappointingly, our efforts to increase the acceptance of the immunization program by providing information in advance were not yet satisfying.
Misconception about pertussis vaccination was prevalent especially among nursing staff. The main reasons for non-compliance included: unawareness of an own risk of infection, the belief that pertussis is not a serious illness, fear of side effects, the belief that the pertussis vaccine might trigger the pertussis disease itself, and skepticism about the efficacy of the pertussis vaccination.
Exposure to locusts, which belong to the arthropod phylum, is an underestimated health problem, especially among workers in research facilities exposed to laboratory animals. We describe a rare case of an occupational immediate-type reaction to locusts with a possible cross-reactivity between desert locust (Schistocerca gregaria) and migratory locust (Locusta migratoria).
Holzgreve, Fabian Christian ; Fräulin, Laura ; Hänel, Jasmin ; Schmidt, Helmut ; Bader, Andreas ; Frei, Markus ; Groneberg, Jan David Alexander ; Ohlendorf, Daniela Maren ; Mark, Anke van
Hintergrund: Dehntrainings sind eine Maßnahme der betrieblichen Gesundheitsförderung (BGF) für Büroangestellte zur Prävention von muskuloskeletalen Erkrankungen (MSE). Sie können zu Beweglichkeitszuwächsen führen und auf psychischer Ebene entspannen. Ziel der Studie war es, ein standardisiertes und individualisiertes Dehntraining am Gerät, das „five-Business“, auf MSE, Lebensqualität und Beweglichkeit zu untersuchen. Dies ist eine Zusammenfassung der international publizierten Ergebnisse.
Methodik: In diese Untersuchung wurden 252 Proband(innen) eingeschlossen, 156 in die Interventionsgruppe (IG), 96 in die Kontrollgruppe (KG). Die IG absolvierte für 12 Wochen 22–24 Trainingseinheiten am „five-Business“, möglichst zweimal wöchentlich. Die Datenerhebung erfolgte mittels sportmotorischer bzw. Range-of-motion(ROM)-Tests, dem Nordic Questionnaire (NQ) und dem SF-36-Fragebogen in Form einer Prä-Post-Untersuchung.
Ergebnisse: Nach der Intervention gaben im NQ signifikant weniger Proband(innen) der IG Beschwerden im oberen Rücken (p < 0,001) im Vergleich zur KG an; keine signifikanten Unterschiede wurden bei Beschwerden im Nacken, Schultern, Hüfte und dem unteren Rücken gefunden. Der mit dem SF-36 erhobene allgemeine Gesundheitszustand und die gesundheitsbezogene Lebensqualität haben sich (nach subjektiven Angaben) signifikant verbessert (Summenscore IG: p = 0,005). In den ROM-Tests waren die Prä-Post-Differenzen bei IG-Proband(innen) in der Sagittalebene (Finger-Boden-Abstand und Retroflexion; p < 0,001) und im modifizierten Schultertest nach Janda auf der linken Seite (p = 0,003) signifikant größer.
Diskussion: Trotz des in Zeitdauer und Häufigkeit relativ geringen Trainingsaufwandes wurden sowohl auf körperlicher als auch auf psychischer Ebene relevante Verbesserungen erzielt, so dass das „five-Business“-Dehntraining als BGF-Maßnahme empfohlen werden kann.
Dreßler, Melanie ; Fußbroich, Daniela ; Böhler, Lisa ; Herrmann, Eva ; Benker, Nicole ; Tytyk, Markus ; Schulze, Johannes B. ; Schubert, Ralf ; Beermann, Christopher ; Zielen, Stefan
Background: Many patients suffering from exercise-induced asthma (EIA) have normal lung function at rest and show symptoms and a decline in FEV1 when they do sports or during exercise-challenge. It has been described that long-chain polyunsaturated fatty acids (LCPUFA) could exert a protective effect on EIA.
Methods: In this study the protective effect of supplementation with a special combination of n-3 and n-6 LCPUFA (sc-LCPUFA) (total 1.19 g/ day) were investigated in an EIA cold air provocation model. Primary outcome measure: Decrease in FEV1 after exercise challenge and secondary outcome measure: anti-inflammatory effects monitored by exhaled NO (eNO) before and after sc-LCPUFA supplementation versus placebo.
Results: Ninety-nine patients with exercise-induced symptoms aged 10 to 45 were screened by a standardized exercise challenge in a cold air chamber at 4 °C. Seventy-three patients fulfilled the inclusion criteria of a FEV1 decrease > 15% and were treated double-blind placebo-controlled for 4 weeks either with sc-LCPUFA or placebo. Thirty-two patients in each group completed the study. Mean FEV1 decrease after cold air exercise challenge and eNO were unchanged after 4 weeks sc-LCPUFA supplementation.
Conclusion: Supplementation with sc-LCPUFA at a dose of 1.19 g/d did not have any broncho-protective and anti-inflammatory effects on EIA.
Trial registration: Clinical trial registration number: NCT02410096. Registered 7 February 2015 at Clinicaltrial.gov
Bladder cancer patients whose tumors develop resistance to cisplatin-based chemotherapy often turn to natural, plant-derived products. Beneficial effects have been particularly ascribed to polyphenols, although their therapeutic relevance when resistance has developed is not clear. The present study evaluated the anti-tumor potential of polyphenol-rich olive mill wastewater (OMWW) on chemo-sensitive and cisplatin- and gemcitabine-resistant T24, RT112, and TCCSUP bladder cancer cells in vitro. The cells were treated with different dilutions of OMWW, and tumor growth and clone formation were evaluated. Possible mechanisms of action were investigated by evaluating cell cycle phases and cell cycle-regulating proteins. OMWW profoundly inhibited the growth and proliferation of chemo-sensitive as well as gemcitabine- and cisplatin-resistant bladder cancer cells. Depending on the cell line and on gemcitabine- or cisplatin-resistance, OMWW induced cell cycle arrest at different phases. These differing phase arrests were accompanied by differing alterations in the CDK-cyclin axis. Considerable suppression of the Akt-mTOR pathway by OMWW was observed in all three cell lines. Since OMWW blocks the cell cycle through the manipulation of the cyclin-CDK axis and the deactivation of Akt-mTOR signaling, OMWW could become relevant in supporting bladder cancer therapy.
Shalamova, Lyudmila ; Felgenhauer, Ulrike ; Wilhelm, Jochen ; Schaubmar, Andreas R. ; Büttner, Kathrin ; Schoen, Andreas ; Widera, Marek ; Ciesek, Sandra ; Weber, Friedemann
The new variant of concern (VOC) of SARS-CoV-2, Omicron (B.1.1.529), is genetically very different from other VOCs. We compared Omicron with the preceding VOC Delta (B.1.617.2) and the wildtype (wt) strain (B.1) with respect to their interactions with the antiviral interferon (IFN-alpha/beta) response in infected cells. Our data indicate that IFN induction by Omicron is low and comparable to the wt, whereas Delta showed an increased IFN induction. However, Omicron exceeded both the wt and the Delta strain with respect to the ability to withstand the antiviral state imposed by IFN-alpha.
Bojkova, Denisa ; Bechtel, Marco ; Rothenburger, Tamara ; Kandler, Joshua D. ; Hayes, Lauren ; Olmer, Ruth Maria ; Martin, Ulrich ; Jonigk, Danny David ; Ciesek, Sandra ; Wass, Mark N. ; Michaelis, Martin ; Cinatl, Jindrich
Recent findings in permanent cell lines suggested that SARS-CoV-2 Omicron BA.1 induces a stronger interferon response than Delta. Here, we show that BA.1 and BA.5 but not Delta induce an antiviral state in air-liquid interface (ALI) cultures of primary human bronchial epithelial (HBE) cells and primary human monocytes. Both Omicron subvariants caused the production of biologically active type I (α/β) and III (λ) interferons and protected cells from super-infection with influenza A viruses. Notably, abortive Omicron infection of monocytes was sufficient to protect monocytes from influenza A virus infection. Interestingly, while influenza-like illnesses surged during the Delta wave in England, their spread rapidly declined upon the emergence of Omicron. Mechanistically, Omicron-induced interferon signalling was mediated via double-stranded RNA recognition by MDA5, as MDA5 knock-out prevented it. The JAK/ STAT inhibitor baricitinib inhibited the Omicron-mediated antiviral response, suggesting it is caused by MDA5-mediated interferon production, which activates interferon receptors that then trigger JAK/ STAT signalling. In conclusion, our study 1) demonstrates that only Omicron but not Delta induces a substantial interferon response in physiologically relevant models, 2) shows that Omicron infection protects cells from influenza A virus super-infection, and 3) indicates that BA.1 and BA.5 induce comparable antiviral states.
On the current psychotherapeutic situation for persons with pornography use disorder in Germany
(2023)
Markert, Charlotte ; Storz, Florian ; Golder, Sarah ; Rechmann, Johanna ; Rubel, Julian ; Lalk, Christopher ; Vogt, Rabea ; Glombiewski, Julia Anna ; Braun, David ; Pané-Farré, Christiane A. ; Michael, Tanja ; Mattheus, Hannah K. ; Dominick, Nanne ; Wölfling, Klaus ; Lutz, Wolfgang ; Schaffrath, Jana ; Stangier, Ulrich ; Kananian, Schahryar ; Strüwing, Dirk ; Klingelhöfer, Doerthe ; Valbert, Frederik Tim ; Neumann, Anja ; Walter, Bertram ; Stark, Rudolf
Background and aims: For the first time, the ICD-11 provides the diagnosis compulsive sexual behavior disorder (CSBD) that can be assigned for pornography use disorder (PUD). This study aimed to estimate the prevalence of PUD and associated consequences in Germany, to identify the psychotherapy demand among likely PUD (lPUD) cases and the treatment supply in different psychotherapeutic settings, to survey psychotherapists' level of expertise regarding PUD, and to identify predictors for psychotherapy demand.
Methods: Four studies were conducted: 1. Online study in the general population (n = 2070; m = 48.9%, f = 50.8%, d = 0.2%), 2. Survey among practicing psychotherapists (n = 983), 3. Survey of psychotherapists in psychotherapeutic outpatient clinics (n = 185), 4. Interviews with psychotherapeutic inpatient clinics (n = 28).
Results: The estimated prevalence of lPUD in the online study was 4.7% and men were 6.3 times more often affected than women. Compared to individuals without PUD, individuals with lPUD more often indicated negative consequences in performance-related areas. Among lPUD cases, 51.2% of men and 64.3% of women were interested in a specialized PUD treatment. Psychotherapists reported 1.2%–2.9% of lPUD cases among their patients. 43.2%–61.5% of psychotherapists stated to be poorly informed about PUD. Only 7% of psychotherapeutic inpatient clinics provided specific treatments to patients with PUD. While, among other factors, negative consequences attributed to lPUD were predictive for psychotherapy demand, weekly pornography consumption, subjective well-being, and religious attachment were not.
Discussion and conclusions: Although PUD occurs quite often in Germany, availability of mental health care services for PUD is poor. Specific PUD treatments are urgently needed.
1. Locating an optimal oviposition site can be a challenging task for female insects, especially when dealing with a patchy, unpredictable and ephemeral food source such as carrion. Understanding the biotic and abiotic parameters that influence the oviposition behaviour of necrophagous flies is not just of great biological importance but also essential for their application in legal investigations.
2. In this study, we monitored the oviposition activity of necrophagous flies (Calliphoridae, Sarcophagidae) using mouse carcasses in an urban (city) and a rural (mixed forest) habitat in Frankfurt/Germany over a 2-year period.
3. Over 240 sampling days, 220,963 larvae of 4 blow fly species and 1 flesh fly were sampled. The most abundant species was the blow fly Lucilia ampullacea, followed by its family members Calliphora vicina and Lucilia caesar, the flesh fly Sarcophaga caerulescens and Lucilia sericata. Up to seven environmental parameters were statistically significant predictors for a colonisation of the carcasses, leading to unique patterns of seasonal and daily oviposition activity for all five species.
4. Overall, the analysis showed that the seasonal adaption (the phenology of each species), the habitat (rural vs. urban) as well as temperature are the most important factors influencing the oviposition behaviour and activity of necrophagous blow flies and flesh flies.
Background
Cochlear Implants (CIs) provide near normal speech intelligibility in quiet environments to individuals suffering from sensorineural hearing loss. Perception of speech in situations with competing background noise and especially music appraisal however are still insufficient. Hence, improving speech perception in ambient noise and music intelligibility is a core challenge in CI research. Quantitatively assessing music intelligibility is a demanding task due to its inherently subjective nature. However, previous approaches have related electrophysiological measurements to speech intelligibility, a corresponding relation to music intelligibility, can be assumed. Recent studies have investigated the relation between results obtained from hearing performance tests and Spread of Excitations (SoEs) measurements. SoE functions are acquired by measuring Electrically Evoked Compound Action Potentials (ECAPs) which represent the electrical response generated in the neural structures of the auditory nerve. The parameters designed to describe SoE functions are used to estimate the dispersal of the electric field in the cochlea. The quality of spatial separation of the electrical field generated by adjacent electrodes are assumed to correlate with hearing performance measures.
Aim of study
This study investigated the relation of parameters derived by ECAP measurements and perceptive skills which aim to access the level of speech and music intelligibility in CI users. In addition, the ratings assessed in a questionnaire on self-rated music intelligibility were correlated to a test battery consisting of measures for speech reception threshold (SRT) in noise (Oldenburger Satztest (OLSA)) and music intelligibility (Adaptive Melody-Pattern-Discrimination Test (AMPDT)). We hypothesised that results from this test battery correlated to subjective ratings and measures describing SoE functions.
Methods
The patient collective covered 17 well-experienced bilateral CI listeners (8 females, 9 males) between the age of 14 and 77 years with a minimum CI experience of two years. Music enjoyment and self-rated musicality was evaluated by means of a questionnaire. The AMPDT included two psychoacoustic tests: timbre difference discrimination threshold (TDDT) and background contour discrimination threshold (BCDT). The accentuation of harmonics in a foreground melody created a background melody. Accentuation was realised by sound level increment, frequency detuning and onset asynchrony. Subjects had to detect target intervals comprising both foreground and background melody by discriminating timbre differences in a Three-Interval Three-Alternative Forced-Choice (3I3AFC) procedure. In a One-Interval Two-Alternative Forced-Choice (1I2AFC) procedure, subjects had to classify the background melody’s contour. SoE was measured via a spatial forward-masking paradigm. A basal, medial and apical recording electrode was measured. Probe electrodes were one electrode position apical to the recording. The width of normalised SoE functions was calculated at their 25 % and 50 % level (excitation distance (DIST)). Furthermore, exponential functions were calculated for SoE profiles with more than three data points for each side. The OLSA assessed SRT in noise. The noisy environment was presented through an array of four loudspeakers (MSNF). The Fastl noise-condition allows to make use of gap listening representing the temporal characteristics of speech as a fluctuating noise. The OLnoise-condition is a continuous noise resulting in a maximum portion of masking.
Results
We found that background melody contour classiffication (BCDT) is more challenging to CI users than the detection of small perceptual timbre differences (TDDT). Background melody contour classification was possible with harmonic accentuation by sound level increment whereas accentuation by onset asynchrony was more demanding. CI users failed in background melody contour classification obtained by frequency detuning. SRTs assessed in the OLSA were significantly lower in the OLnoise than in the Fastl noise masking condition. A number of N = 90 SoE functions were acquired from ECAP measurements, in which N = 48 showed a clearly present ECAP response. The DIST at the 25 % and 50 % level was narrower for the basal than for the apical and medial electrode. SoE functions showed asymmetric profiles with larger amplitudes towards the basal end of the cochlea. Correlation analysis between the AMPDT, OLSA and DISTs showed no significant correlation. Correlation analysis between the AMPDT, OLSA and the questionnaire’s results could not prove that musical activities (music listening, singing or playing instruments) improve music intelligibility. However, CI supply has restored the importance of music, self-rated musicality and musical enjoyment in this study’s subjects.
Conclusions
The present study’s results imply that CI listeners are only able to detect distinct timbre alterations throughout the course of a musical piece whereas they cannot discriminate background melodies hidden in a pattern of complex harmonic sounds. Furthermore, SoE measurements do not seem to be an adequate tool to predict neither speech nor music intelligibility in CI listeners, contrary to our initial hypothesis. This finding is consistent with a number of studies who did not find a correlation between music or speech intelligibility and channel interactions assessed by SoE measurements. It can be concluded that albeit CI supply restores musical enjoyment in patients with sensorineural hearing loss, music perception is still poor and does not significantly improve by regular musical activities such as listening to music, singing or playing instruments.
Dávidová, Petra ; Müller, Michael ; Wenner, Yaroslava ; König, Clara ; Kenikstul, Ninel ; Kohnen, Thomas
Purpose: Filler injections for aesthetic purposes are very popular, but can have far-reaching and irreversible consequences. This report describes the course of a patient with devastating complications after glabellar hyaluronic acid injection, their pathomechanism, management and outcome.
Observations: A healthy, 43-year-old woman underwent her first hyaluronic acid injection in the glabella and went blind on her left eye immediately thereafter. Massaging of the injection area and observation were performed, before she presented with swelling of the left forehead and upper lid, ptosis, complete ophthalmoplegia and blindness in our hospital. Immediate massaging of the globe and systemic therapy including acetylsalicylic acid, tinzaparin sodium and cortisone was initiated and hyaluronidase injections in the injection area were performed. In the further course, the patient developed necrotic and hemorrhagic skin and mucosal lesions, lagophthalmos, anterior and posterior segment ischemia and globe hypotonia with consecutive globe deformation. In the follow-up of 2.5 months, lid swelling, lagophthalmos and ptosis resolved and keratopathy improved but blindness, skin lesions and strabismus with reduced eye motility were still present and madarosis and early enophthalmos were detected.
Conclusions and Importance: The outcome of ophthalmic artery occlusion after hyaluronic acid filler injection is poor. Sufficient knowledge about facial anatomy, the implementation of filler injections and the management of complications is essential for the practitioner. The patient should be clarified about potential and even rare risks of these procedures.
Optimal distribution-preserving downsampling of large biomedical data sets (opdisDownsampling)
(2021)
Motivation: The size of today’s biomedical data sets pushes computer equipment to its limits, even for seemingly standard analysis tasks such as data projection or clustering. Reducing large biomedical data by downsampling is therefore a common early step in data processing, often performed as random uniform class-proportional downsampling. In this report, we hypothesized that this can be optimized to obtain samples that better reflect the entire data set than those obtained using the current standard method. Results: By repeating the random sampling and comparing the distribution of the drawn sample with the distribution of the original data, it was possible to establish a method for obtaining subsets of data that better reflect the entire data set than taking only the first randomly selected subsample, as is the current standard. Experiments on artificial and real biomedical data sets showed that the reconstruction of the remaining data from the original data set from the downsampled data improved significantly. This was observed with both principal component analysis and autoencoding neural networks. The fidelity was dependent on both the number of cases drawn from the original and the number of samples drawn. Conclusions: Optimal distribution-preserving class-proportional downsampling yields data subsets that reflect the structure of the entire data better than those obtained with the standard method. By using distributional similarity as the only selection criterion, the proposed method does not in any way affect the results of a later planned analysis.
In der vorliegenden Arbeit wird ein quantitatives Verfahren zur Auswertung des ELI S As vorgestellt das alle Anforderungen an eine gute Antikörpermessung erfüllt und sich besonders durch große klinische Plausibilität und Ökonomie in der Virusdiagnostik auszeichnet. Die Forderung nach der überregionalen Teststandardisierung wird berücksichtigt. Verschiedene Methoden zur quantitativen Antikörpermessung werden verglichen. Bestimmt werden Antikörper der Immunglobulinklasse G (IgG) gegen das Zytomegalievirus (CMV). Die Methode basiert auf der Herstellung einer einzigen Serumverdünnung (1:160) und führt in Kombination mit einer Standardkurve zu Ergebnissen in Form von Titern. Drei Varianten mit unterschiedlichen Anwendungsschwerpunkten stehen zur Verfügung. Für jedes Probanden-Serum wird immer die Reaktion mit Antigen und Kontrollantigen gleichzeitig in einem Testansatz geprüft.
Acute lymphoblastic leukemia (ALL) is a malignancy of lymphoid progenitor cells occurring at an annual incidence rate of approximately 1.1 to 2.1 per 100,000 person-years globally. Approximately 40% of annual ALL cases occur in adults, yet estimated 5-year overall survival rates are about 40% to 50% in adults (and vary broadly by age) compared with 90% in children. Although the addition and/or intensification of asparaginase as a key treatment strategy for pediatric ALL is well recognized, further research is needed to clarify the benefit/risk ratio in adult patients with ALL. This review emphasizes the importance of efficient management of adverse events to increase asparaginase efficacy and explores novel strategies for optimizing asparaginase treatment, including new formulations of asparaginase, pharmacokinetic-based dosing, and pharmacogenetic profiling. Upcoming results of adult ALL trials should further clarify the role of asparaginase, building on the results of the large NOPHO 2008, CALGB 10403, GRAALL-2005, GMALL 07/2003, and UKALL14 trials.
Ahmadi, Salim ; Klingelhöfer, Doris ; Erbe, Christina ; Holzgreve, Fabian Christian ; Groneberg, Jan David Alexander ; Ohlendorf, Daniela Maren
Objectives: Inadequate oral hygiene still leads to many serious diseases all over the world. Therefore, this study aimed to analyze scientific research in the field of oral health in order to be able to comprehend their relevant subject areas, research connections, or developments. Methods: This study aimed to assess the global publication output on oral hygiene to create a world map that provides background information on key players, trends, and incentives of research. For this purpose, established bibliometric parameters were combined with state-of-the-art visualization techniques. Results: This study shows the actual key players of research on oral hygiene in high-income economies with only marginal participation from lower economies. This still corresponds to the current burden situations, but they are more and more shifting to the disadvantage of the low-income countries. There is a clear North–South and West–East gradient, with the USA and the Western European nations being the most publishing nations on oral hygiene. As an emerging country, Brazil plays a role in the research. Conclusions: The scientific power players were concentrated in high-income countries. However, the changing epidemiological situation requires a different scientific approach to oral hygiene. This requires an expansion of the international network to meet the demands of future global oral health burdens, which are mainly related to oral hygiene.
Römer, Thomas ; Bitzer, Johannes ; Egarter, Christian ; Hadji, Peyman ; Kiechle, Marion ; Kramer, Heike ; Oppelt, Patricia G. ; Peters, Klaus H. ; Stute, Petra ; Schaudig, Katrin ; Wiegratz, Inka ; Regidor, Pedro-Antonio
Hormonal contraceptives are an effective and safe method for preventing pregnancy. Progestins used in contraception are either components of combined hormonal contraceptives (tablets, patches or vaginal rings) or are used as a single active ingredient in progestin mono-preparations (the progestin-only pill (POP), implants, intrauterine systems or depot preparations). Progestins are highly effective in long-term contraception when used properly, and have a very good safety profile with very few contraindications. A new oestrogen-free ovulation inhibitor (POP) has recently been authorised in the USA and the EU. This progestin mono-preparation contains 4 mg of drospirenone (DRSP), which has anti-gonadotropic, anti-mineralocorticoidic and anti-androgenic properties. The hormone administration regimen of 24 days followed by a 4-day hormone-free period was chosen to improve bleeding control and to maintain oestradiol concentrations at early follicular-phase levels, preventing oestrogen deficiency. Clinical trials have demonstrated a high contraceptive effectiveness, a very low risk of cardiovascular side effects and a favourable menstrual bleeding pattern. Due to the long half-life of DRSP (30 – 34 hours), the effectiveness of the preparation is maintained even if a woman forgets to take a pill on a single occasion. Studies involving deliberate 24-hour delays in taking a pill have demonstrated that ovulation inhibition is maintained if a single pill is missed. Following a summary of the current status of oestrogen-free contraception, this review article will describe the clinical development programme of the 4 mg DRSP mono-preparation and the resulting data on the effectiveness and safety of this new oestrogen-free oral hormonal contraceptive.
Seifert, Lukas Benedikt ; Mainka, Tim ; Herrera-Vizcaino, Carlos ; Verboket, René ; Sader, Robert Alexander
Objective: The aim of this study was to retrospectively review the midface and orbital floor fractures treated at our institution with regard to epidemiological aspects, surgical treatment options and postoperative complications and discuss this data with the current literature. Study design: One thousand five hundred and ninety-four patients with midface and orbital fractures treated at the Department of Oral, Cranio-Maxillofacial and Facial Plastic Surgery of the Goethe University Hospital in Frankfurt (Germany) between 2007 and 2017 were retrospectively reviewed. The patients were evaluated by age, gender, etiology, fracture pattern, defect size, surgical treatment and complications. Results: The average patient age was 46.2 (± 20.8). Most fractures (37.5%) occurred in the age between 16 and 35. Seventy-two percent of patients were male while 28% were female. The most common cause of injury was physical assault (32.0%) followed by falls (30.8%) and traffic accidents (17.0%). The average orbital wall defect size was 297.9 mm2 (± 190.8 mm2). For orbital floor reconstruction polydioxanone sheets (0.15 mm 38.3%, 0.25 mm 36.2%, 0.5 mm 2.8%) were mainly used, followed by titanium meshes (11.5%). Reconstructions with the 0.15 mm polydioxanone sheets showed the least complications (p < 0.01, r = 0.15). Eighteen percent of patients who showed persistent symptoms and post-operative complications: 12.9% suffered from persistent hypoesthesia, 4.4% suffered from post-operative diplopia and 3.9% showed intra-orbital hematoma. Conclusion: Results of the clinical outcome in our patients show that 0.15 mm resorbable polydioxanone sheets leads to significantly less post-operative complications for orbital floor defects even for defects beyond the recommended 200 mm2.
Sander, Myriam Christine ; Fandakova, Yana ; Grandy, Thomas H. ; Shing, Yee Lee ; Werkle-Bergner, Markus
We studied oscillatory mechanisms of memory formation in 48 younger and 51 older adults in an intentional associative memory task with cued recall. While older adults showed lower memory performance than young adults, we found subsequent memory effects (SME) in alpha/beta and theta frequency bands in both age groups. Using logistic mixed effects models, we investigated whether interindividual differences in structural integrity of key memory regions could account for interindividual differences in the strength of the SME. Structural integrity of inferior frontal gyrus (IFG) and hippocampus was reduced in older adults. SME in the alpha/beta band were modulated by the cortical thickness of IFG, in line with its hypothesized role for deep semantic elaboration. Importantly, this structure–function relationship did not differ by age group. However, older adults were more frequently represented among the participants with low cortical thickness and consequently weaker SME in the alpha band. Thus, our results suggest that differences in the structural integrity of the IFG contribute not only to interindividual, but also to age differences in memory formation.