Adaptierte Version des Planspiels Energetingen für Hauptschulen

Ein Projektbericht mit Materalien für die Umsetzun

Pan-Bcl-2 inhibitor Obatoclax is a potent late stage autophagy inhibitor in colorectal cancer cells independent of canonical autophagy signaling

Background: Colorectal cancer is the third most common malignancy in humans and novel therapeutic approaches are urgently needed. Autophagy is an evolutionarily highly conserved cellular process by which cells collect unnecessary organelles or misfolded proteins and subsequently degrade them in vesicular structures in order to refuel cells with energy. Dysregulation of the complex autophagy signaling network has been shown to contribute to the onset and progression of cancer in various models. The Bcl-2 family of proteins comprises central regulators of apoptosis signaling and has been linked to processes involved in autophagy. The antiapoptotic members of the Bcl-2 family of proteins have been identified as promising anticancer drug targets and small molecules inhibiting those proteins are in clinical trials. Methods: Flow cytometry and colorimetric assays were used to assess cell growth and cell death. Long term 3D cell culture was used to assess autophagy in a tissue mimicking environment in vitro. RNA interference was applied to modulate autophagy signaling. Immunoblotting and q-RT PCR were used to investigate autophagy signaling. Immunohistochemistry and fluorescence microscopy were used to detect autophagosome formation and autophagy flux. Results: This study demonstrates that autophagy inhibition by obatoclax induces cell death in colorectal cancer (CRC) cells in an autophagy prone environment. Here, we demonstrate that pan-Bcl-2 inhibition by obatoclax causes a striking, late stage inhibition of autophagy in CRC cells. In contrast, ABT-737, a Mcl-1 sparing Bcl-2 inhibitor, failed to interfere with autophagy signaling. Accumulation of p62 as well as Light Chain 3 (LC3) was observed in cells treated with obatoclax. Autophagy inhibition caused by obatoclax is further augmented in stressful conditions such as starvation. Furthermore, our data demonstrate that inhibition of autophagy caused by obatoclax is independent of the essential pro-autophagy proteins Beclin-1, Atg7 and Atg12. Conclusions: The objective of this study was to dissect the contribution of Bcl-2 proteins to autophagy in CRC cells and to explore the potential of Bcl-2 inhibitors for autophagy modulation. Collectively, our data argue for a Beclin-1 independent autophagy inhibition by obatoclax. Based on this study, we recommend the concept of autophagy inhibition as therapeutic strategy for CRC

Persistence of psychological distress in surgical patients with interest in psychotherapy: results of a 6-month follow-up.

OBJECTIVES: This prospective observational study investigated whether self-reported psychological distress and alcohol use problems of surgical patients change between preoperative baseline assessment and postoperative 6-month follow-up examination. Patients with preoperative interest in psychotherapy were compared with patients without interest in psychotherapy. METHODS: A total of 1,157 consecutive patients from various surgical fields completed a set of psychiatric questionnaires preoperatively and at 6 months postoperatively, including Patient Health Questionnaire-4 (PHQ-4), Brief Symptom Inventory (BSI), Center for Epidemiologic Studies Depression Scale (CES-D), World Health Organization 5-item Well-Being Index (WHO-5), and Alcohol Use Disorder Identification Test (AUDIT). Additionally, patients were asked for their interest in psychotherapy. Repeated measure ANCOVA was used for primary data analysis. RESULTS: 16.7% of the patients were interested in psychotherapy. Compared to uninterested patients, they showed consistently higher distress at both baseline and month 6 regarding all of the assessed psychological measures (p's between <0.001 and 0.003). At 6-month follow-up, neither substantial changes over time nor large time x group interactions were found. Results of ANCOVA's controlling for demographic variables were confirmed by analyses of frequencies of clinically significant distress. CONCLUSION: In surgical patients with interest in psychotherapy, there is a remarkable persistence of elevated self-reported general psychological distress, depression, anxiety, and alcohol use disorder symptoms over 6 months. This suggests high and chronic psychiatric comorbidity and a clear need for psychotherapeutic and psychiatric treatment rather than transient worries posed by facing surgery

6-month follow-up of psychological distress and alcohol use problems of surgical patients with interest in psychotherapy (n = 193) and surgical patients without interest in psychotherapy (n = 964).⁺

+<p>Repeated measures ANCOVA; estimated marginal means (EMM) and standard error of the mean (SEM); statistical significance of Bonferroni correction: p<0.0041; T1 (preoperative baseline assessment) and T2 (postoperative 6 months follow-up) as within-subject factors, therapy interest as between subject factor; covariate: age; additional between-subject factors: gender and partnership status.</p>++<p>Number ranges for the specific variables from 173 to 187 (patients interested in therapy contacts), and from 878 to 938 (patients not interested in therapy contacts) because of missing data</p>a)<p>BSI: Brief Symptom Inventory; GSI: General severity index</p>b)<p>Visual analogue scale of the EQ-5D, 0 to 100 with higher scores indicating better subjective health</p>c)<p>WHO-5: World Health Organization 5-item Well-Being Index</p>d)<p>PHQ-2: Patient Health Questionnaire-4, depression subscale</p>e)<p>CES-D: Center for Epidemiologic Studies Depression Scale</p>f)<p>GAD-2: Patient Health Questionnaire-4, anxiety subscale</p>g)<p>AUDIT: Alcohol Use Disorder Identification Test, AUDIT-C: AUDIT subscore for score for risky alcohol consumption</p

Comparisons of participants (n = 1,157) and nonparticipants (n = 3,411) of the 6-month follow-up; n (%); mean [SD].

+<p>Number ranges for the specific variables from 1,123 to 1,157 (participants) and from 3,236 to 3,411 (nonparticipants) because of missing data.</p>++<p>Data for ASA and surgical field are available for the implementation phase; numbers account to 715 (participants), and 2266 (nonparticipants) because of missing data.</p>a)<p>ASA (American Society of Anesthesiologists) physical status classification: (I) Healthy patient; (II) Mild systemic disease, no functional limitation; (III) Severe systemic disease with definite functional limitation; (IV) Severe systemic disease that is a constant threat to life;</p>b)<p>BSI: Brief Symptom Inventory; GSI: General severity index;</p>c)<p>Visual analogue scale of the EQ-5D, 0 to 100 with higher scores indicating better subjective health;</p>d)<p>WHO-5: World Health Organization 5-item Well-Being Index;</p>e)<p>PHQ-2: Patient Health Questionnaire-4, depression subscale;</p>f)<p>CES-D: Center for Epidemiologic Studies Depression Scale;</p>g)<p>GAD-2: Patient Health Questionnaire-4, anxiety subscale;</p>h)<p>AUDIT: Alcohol Use Disorder Identification Test, AUDIT-C: AUDIT subscore for risky alcohol consumption.</p

6-month follow-up of rates of clinically significant distress and alcohol use problems of surgical patients with interest in psychotherapy (n = 193) and surgical patients without interest in psychotherapy (n = 964).⁺

+<p>McNemar’s test; T1: Preoperative baseline assessment; T2: Postoperative 6 months follow-up; n (%); statistical significance of Bonferroni correction: p<0.0045.</p><p>A case with clinically significant distress was defined as a patient scoring above the cut off score of a given questionnaire (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0051167#pone-0051167-t001" target="_blank">Table 1</a> for cut off scores of all measures).</p>++<p>Number ranges for the specific variables from 185 to 191 (patients interested in therapy contacts), and from 907 to 943 (patients not interested in therapy contacts) because of missing data.</p>a)<p>BSI: Brief Symptom Inventory; GSI: General severity index;</p>b)<p>WHO-5: World Health Organization 5-item Well-Being Index;</p>c)<p>PHQ-2: Patient Health Questionnaire-4, depression subscale;</p>d)<p>CES-D: Center for Epidemiologic Studies Depression Scale;</p>e)<p>GAD-2: Patient Health Questionnaire-4, anxiety subscale;</p>f)<p>AUDIT: Alcohol Use Disorder Identification Test, AUDIT-C: AUDIT subscore for score for risky alcohol consumption.</p

Standardized self-report questionnaires that were assessed at baseline (T1) and 6-month follow-up (T2).

<p>Standardized self-report questionnaires that were assessed at baseline (T1) and 6-month follow-up (T2).</p

Sociodemographic and clinical characteristics of all participants of the 6-month follow-up (N = 1,157), as well as comparison of patients who showed interest in psychotherapy (n = 193) and patients who were not interested in psychotherapy (n = 964); mean [SD], n (%).

+<p>Number ranges for the specific variables from 1,145 to 1,157 (all participants), from 189 to 193 (patients interested in psychotherapy) and from 953 to 964 (patients not interested in psychotherapy) because of missing data.</p>++<p>Data for ASA and surgical field are available for the implementation phase; numbers account to 715 (all participants), 101 (patients interested in psychotherapy, and 614 (patients not interested in psychotherapy) because of missing data.</p>a)<p>ASA (American Society of Anesthesiologists) physical status classification: (I) Healthy patient; (II) Mild systemic disease, no functional limitation; (III) Severe systemic disease with definite functional limitation; (IV) Severe systemic disease that is a constant threat to life.</p

Secondary structure determination of conserved SARS-CoV-2 RNA elements by NMR spectroscopy

The current pandemic situation caused by the Betacoronavirus SARS-CoV-2 (SCoV2) highlights the need for coordinated research to combat COVID-19. A particularly important aspect is the development of medication. In addition to viral proteins, structured RNA elements represent a potent alternative as drug targets. The search for drugs that target RNA requires their high-resolution structural characterization. Using nuclear magnetic resonance (NMR) spectroscopy, a worldwide consortium of NMR researchers aims to characterize potential RNA drug targets of SCoV2. Here, we report the characterization of 15 conserved RNA elements located at the 5' end, the ribosomal frameshift segment and the 3'-untranslated region (3'-UTR) of the SCoV2 genome, their large-scale production and NMR-based secondary structure determination. The NMR data are corroborated with secondary structure probing by DMS footprinting experiments. The close agreement of NMR secondary structure determination of isolated RNA elements with DMS footprinting and NMR performed on larger RNA regions shows that the secondary structure elements fold independently. The NMR data reported here provide the basis for NMR investigations of RNA function, RNA interactions with viral and host proteins and screening campaigns to identify potential RNA binders for pharmaceutical intervention