Mehr als ein Papiertiger? : Die Konfliktbearbeitungspolitik der GASP in Afrika

Die Europäische Union und Afrika - jahrelang wurde diese Verbindung mit entwicklungspolitischer Zusammenarbeit im Rahmen der Lomé-Abkommen assoziiert. Die außen- und sicherheitspolitischen Interessen beschränkten sich auf die bilaterale und zumeist postkoloniale Ebene. Erst seit Mitte der neunziger Jahre entstanden in der EU programmatische Entwürfe für eine umfassende Konfliktbearbeitung in Afrika. In der vorliegenden Studie zeichnet die Autorin die Stufen des Afrikakonzepts der EU nach und stellt diesem die tatsächlich umgesetzte Politik der GASP am Beispiel Ruandas und der Demokratischen Republik Kongo gegenüber. Ausgelöst durch das Versagen der internationalen Gemeinschaft angesichts des Völkermords in Ruanda wurden vom Außenministerrat zunächst vorsichtige, dann von der EU-Kommission weitreichende Konzepte für eine Verbindung entwicklungs- und außenpolitischer Ziele der Konfliktbearbeitung formuliert. Doch erweist sich die politische Wirklichkeit - gemessen an ihrem programmatischen Anspruch - als eher defizitär. Zwar haben die Konzepte der EU noch nicht zu einem grundlegenden Wandel auf der Ebene des policy making geführt, doch lassen sich eine gewisse kontinuierliche Strategie und erste Anzeichen einer "Europäisierung" in den Politiken der Gemeinschaft erkennen. Ein solcher gemeinsamer Ansatz würde dabei nicht nur eine effektive Implementierung der GASP, sondern auch eine Profilierung der EU als ernst zu nehmender Konfliktbearbeitungsakteur mit sich bringen

Inhibitor of apoptosis proteins, NAIP, cIAP1 and cIAP2 expression during macrophage differentiation and M1/M2 polarization

Monocytes and macrophages constitute the first line of defense of the immune system against external pathogens. Macrophages have a highly plastic phenotype depending on environmental conditions; the extremes of this phenotypic spectrum are a pro-inflammatory defensive role (M1 phenotype) and an anti-inflammatory tissue-repair one (M2 phenotype). The Inhibitor of Apoptosis (IAP) proteins have important roles in the regulation of several cellular processes, including innate and adaptive immunity. In this study we have analyzed the differential expression of the IAPs, NAIP, cIAP1 and cIAP2, during macrophage differentiation and polarization into M1 or M2. In polarized THP-1 cells and primary human macrophages, NAIP is abundantly expressed in M2 macrophages, while cIAP1 and cIAP2 show an inverse pattern of expression in polarized macrophages, with elevated expression levels of cIAP1 in M2 and cIAP2 preferentially expressed in M1. Interestingly, treatment with the IAP antagonist SMC-LCL161, induced the upregulation of NAIP in M2, the downregulation of cIAP1 in M1 and M2 and an induction of cIAP2 in M1 macrophages.This work was supported by Universidad de Granada, Plan Propio 2015;#P3B: FAM, VMC (http://investigacion.ugr.es/pages/planpropio/2015/ resoluciones/p3b_def_28072015); Universidad de Granada CEI BioTic;#CAEP2-84: VMC (http:// biotic.ugr.es/pages/resolucionprovisional enseaanzapractica22demayo/!); and Canadian nstitutes of Health Research;#231421, #318176, #361847: STB, ECL, RK (http://www.cihr-irsc.gc. ca/e/193.html). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Global overview of the management of acute cholecystitis during the COVID-19 pandemic (CHOLECOVID study)

Background: This study provides a global overview of the management of patients with acute cholecystitis during the initial phase of the COVID-19 pandemic. Methods: CHOLECOVID is an international, multicentre, observational comparative study of patients admitted to hospital with acute cholecystitis during the COVID-19 pandemic. Data on management were collected for a 2-month study interval coincident with the WHO declaration of the SARS-CoV-2 pandemic and compared with an equivalent pre-pandemic time interval. Mediation analysis examined the influence of SARS-COV-2 infection on 30-day mortality. Results: This study collected data on 9783 patients with acute cholecystitis admitted to 247 hospitals across the world. The pandemic was associated with reduced availability of surgical workforce and operating facilities globally, a significant shift to worse severity of disease, and increased use of conservative management. There was a reduction (both absolute and proportionate) in the number of patients undergoing cholecystectomy from 3095 patients (56.2 per cent) pre-pandemic to 1998 patients (46.2 per cent) during the pandemic but there was no difference in 30-day all-cause mortality after cholecystectomy comparing the pre-pandemic interval with the pandemic (13 patients (0.4 per cent) pre-pandemic to 13 patients (0.6 per cent) pandemic; P = 0.355). In mediation analysis, an admission with acute cholecystitis during the pandemic was associated with a non-significant increased risk of death (OR 1.29, 95 per cent c.i. 0.93 to 1.79, P = 0.121). Conclusion: CHOLECOVID provides a unique overview of the treatment of patients with cholecystitis across the globe during the first months of the SARS-CoV-2 pandemic. The study highlights the need for system resilience in retention of elective surgical activity. Cholecystectomy was associated with a low risk of mortality and deferral of treatment results in an increase in avoidable morbidity that represents the non-COVID cost of this pandemic

Omecamtiv mecarbil in chronic heart failure with reduced ejection fraction, GALACTIC‐HF: baseline characteristics and comparison with contemporary clinical trials

Aims: The safety and efficacy of the novel selective cardiac myosin activator, omecamtiv mecarbil, in patients with heart failure with reduced ejection fraction (HFrEF) is tested in the Global Approach to Lowering Adverse Cardiac outcomes Through Improving Contractility in Heart Failure (GALACTIC‐HF) trial. Here we describe the baseline characteristics of participants in GALACTIC‐HF and how these compare with other contemporary trials. Methods and Results: Adults with established HFrEF, New York Heart Association functional class (NYHA) ≥ II, EF ≤35%, elevated natriuretic peptides and either current hospitalization for HF or history of hospitalization/ emergency department visit for HF within a year were randomized to either placebo or omecamtiv mecarbil (pharmacokinetic‐guided dosing: 25, 37.5 or 50 mg bid). 8256 patients [male (79%), non‐white (22%), mean age 65 years] were enrolled with a mean EF 27%, ischemic etiology in 54%, NYHA II 53% and III/IV 47%, and median NT‐proBNP 1971 pg/mL. HF therapies at baseline were among the most effectively employed in contemporary HF trials. GALACTIC‐HF randomized patients representative of recent HF registries and trials with substantial numbers of patients also having characteristics understudied in previous trials including more from North America (n = 1386), enrolled as inpatients (n = 2084), systolic blood pressure < 100 mmHg (n = 1127), estimated glomerular filtration rate < 30 mL/min/1.73 m2 (n = 528), and treated with sacubitril‐valsartan at baseline (n = 1594). Conclusions: GALACTIC‐HF enrolled a well‐treated, high‐risk population from both inpatient and outpatient settings, which will provide a definitive evaluation of the efficacy and safety of this novel therapy, as well as informing its potential future implementation

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Condensin complex contributes to VSG expression, regulation and switching in Trypanosoma brucei

African trypanosomes evade the host immune response by periodically changing their protein coat, constituted by a single Variant Surface Glycoprotein (VSG), allowing for chronic infections. We have previously published that Cohesin complex regulates in situ transcriptional VSG switching, as partial depletion of Cohesin subunits increases the frequency of antigenic variation. Condensin complex, structurally similar to the cohesin complex, has recently emerged in eukaryotes as a mayor regulator of chromosome architecture, chromatin compaction during interphase, and is greatly enriched near highly expressed genes. Previous results suggest that SUMOylation of chromatin at the active VSG locus may function to nucleate factors to the nuclear body ESB, where VSG transcription occurs. Furthermore, we found by proteomic analysis that TbSMC4, a subunit of the condensin complex, is a consistent and abundant SUMO target. Therefore, it seems probable that Condensin functions in the regulation of VSG monoallelic expression and/or transcriptional switching. Co-IP experiments showed that trypanosome condensin includes well-known subunits CND1 & 2 and SMC4, suggesting a conserved multiprotein complex that localizes in the nucleus, as described in other eukaryotes. In addition, we found distinct SUMOylation sites in the condensin subunits of the infective bloodstream forms. Interestingly, partial depletion of condensin subunits resulted in a significant increase of VSG221 switching off events, reaching up to 10% of the population, a switching frequency higher than previously described for cohesin depletion. Isolated switches corresponded to in situ transcriptional activation events of independent telomeric VSG-ESs. All data suggest that condensin complex has a key role in establishing and/or maintaining the transcriptional state of VSG-ES chromatin

Establishment of a structure-activity relationship of the 1H-imidazo[4,5-c]quinoline-based kinase inhibitor NVP-BEZ235 as a lead for African sleeping sickness

The compound NVP-BEZ235 (1) is a potent inhibitor of human phospoinositide-3-kinases (PI3Ks) and mammalian target of rapamycin (mTOR) that also showed high inhibitory potency against T. brucei cultures. With an eye towards using 1 as a starting point for anti-trypanosomal drug discovery, we report efforts to reduce host cell toxicity, improve the physicochemical properties and improve the selectivity profile over human kinases. In this work we have developed structure-activity relationships (SAR) for analogs of 1 and have prepared analogs of 1 with improved solubility properties and good predicted central nervous system exposure. In that way, we have identified 4e, 9, 16e and 16g as the most promising leads to date. We also report cell phenotype and phospholipidomic studies that suggest that these compounds exert their anti-trypanosomal effects, at least in part, by inhibition of lipid kinases.Publisher PDFPeer reviewe

SUMOylation of chromatin-associated proteins occurs at the active <i>VSG-</i>ES in the bloodstream form.

<p>(A) Schema of tagged cell lines used in ChIP experiments. SALR cell line: <i>firefly luciferase</i> gene (FLuc) inserted downstream of the active <i>VSG221</i>-ES promoter and SILR: FLuc gene inserted downstream of an inactive <i>VSG</i>-ES. <i>Renilla</i>-luciferase reporter gene (RLuc) was inserted in the tubulin locus of both cell lines, as RNA pol II control. Fragments amplified by qPCR using ChIPed DNA are indicated (primers are listed in the <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004545#ppat.1004545.s010" target="_blank">Table S1</a>). (B), (C) TbRPA1 ChIP analysis in SALR and SILR shows the occupancy of RNA pol I in these cell lines. High TbRPA1 enrichment is found in SALR along the active <i>VSG</i>-ES at FLuc gene, pseudo-VSG (pseVSG) and the active <i>VSG221</i> in contrast to inactive <i>VSGs</i>. Similar occupancy is detected in SILR except for the FLuc gene inserted in an inactive <i>VSG-ES</i> promoter of the BES5 in this cell line. The RNA pol I occupancies between FLuc SALR and FLuc SILR were found significantly different (p-value<0.01). Sequences present in rDNA promoter and 18S gene transcribed by RNA pol I were analyzed as positive controls and RNA pol II or pol III transcribed genes as negative controls. (D), (E) ChIP analysis using anti-TbSUMO mAb in SALR and SILR cell lines. Enrichment of SUMOylated proteins is found at the active <i>VSG</i>-ES chromatin from the promoter to the telomeric <i>VSG221</i>. SUMO ChIP levels between active (FLuc SALR) and inactive (FLuc SILR) reporters are significantly different (p-value<0.01). Similarly, SUMO enrichment on the active <i>VSG221</i> versus the inactive <i>VSGs</i> (<i>121</i>, <i>JS1</i> and <i>VO2</i>) is significantly different (p-values<0.05). ChIP analyses show the average from at least three independent experiments with standard error (SE). Data are represented as percentage of input immunoprecipitated (% input) after background subtraction of the negative control ChIP using the pre-bleed antiserum. Tubulin (Tub), Splicer Leader (SL), EP Procyclin promoter (EP pro), Procyclin gene (EP cds), rDNA promoter (rDNA pro), rDNA spacer (rDNA sp).</p

SUMOylation of chromatin-associated proteins by TbSIZ1 is important for efficient recruitment of RNA pol I and <i>VSG</i>-ES expression.

<p>(A) Reduced <i>VSG</i>-ES chromatin SUMOylation upon TbSIZ1 depletion. ChIP analysis carried out in TbSIZ1 depleted cells (48 h) and parental cell line (SALR) shows that SUMOylated chromatin is significantly reduced along all active <i>VSG</i>-ES, from the upstream promoter to the active <i>VSG221</i> gene. (B) Reduced RNA pol I occupancy upon TbSIZ1 depletion. Occupancy of RNA pol I was determined by TbRPA1 ChIP. Statistical analysis shows a significant difference of TbRPA1 levels between parental and TbSIZ1 depleted cells at the active <i>VSG</i>-ES. Data from three independent TbSIZ1 RNAi clones and parental controls are represented as percentage of input immunoprecipitated after background subtraction of the pre-bleed antiserum ChIP. The results show the average from at least three independent experiments with standard error (SE). Statistical analysis (Student's t-test), *p<0.05, **p<0.01. (C) Reduced RNA pol I occupancy results in reduced <i>VSG</i>-ES derived transcripts. Quantitative RT-PCR analysis shows reduced amounts of <i>FLuc</i> reporter gene and <i>VSG221</i> mRNA without significant effect in rDNA transcripts or RNA pol II derived transcripts <i>RLuc</i> and myosin. Results are the average from three independent clones. Data were normalized with U2 mRNA, transcribed by RNA pol III. Statistical analysis (Student's t-test) *p<0.05, **p<0.01.</p