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Background: Point-of-care tests (POCTs) for C-reactive protein can support clinical decision-making of general practitioners (GPs) but are not widely used in German general practices., Aim: To investigate the utilization of semi-quantitative CRP-POCTs in routine primary care., Design & Setting: Prospective observational study in 49 general practices in Germany (Nov/2022 to Apr/2023)., Method: GPs were provided with CRP-POCTs and collected data for each CRP-POCT use using standardized data collection sheets., Results: Data from 1,740 CRP-POCT uses were recorded. GPs employed CRP-POCTs mainly for patients with respiratory tract infections (RTIs, 70.9% of all cases) and to a lesser extent for gastrointestinal infections (GIs, 10.3%). In RTIs, CRP-POCTs were frequently used to distinguish between bacterial and viral aetiology (60.8%) and to guide decisions on antibiotic prescribing (62.8%). In GIs, CRP-POCTs were mainly used to rule out severe disease progressions (53.2%) and for decisions on further diagnostic procedures (45.6%). In RTIs, CRP-POCTs influenced antibiotic prescribing in 77.5 % of the cases (32.3% in favour vs . 45.2% waiver). In GIs, CRP levels mainly affected decisions on further diagnostic procedures. GPs reported that CRP-POCTs were helpful in 88.6% of all cases., Conclusions: When available, German GPs predominantly use semi-quantitative CRP-POCTs to guide decisions on antibiotic prescribing in patients with RTI. CRP-POCT use improves clinical decision-making and increases the GPs' clinical confidence., (Copyright © 2024, The Authors.)

Objectives: Out-of-hours outpatient emergency medical services (OEMS) provide healthcare for patients with non-life-threatening conditions in need for urgent care when outpatient practices are closed. We studied the use of point-of-care-testing of C-reactive protein (CRP-POCT) at OEMS., Design: Cross-sectional questionnaire-based survey., Setting: Single centre OEMS practice in Hildesheim, Germany (October 2021 to March 2022)., Participants: OEMS physicians answering a questionnaire immediately after performing CRP-POCTs (CUBE-S Analyzer, Hitado) on any patients., Primary and Secondary Outcomes: Impact of CRP-POCTs on clinical decision-making and perceived usefulness., Results: In the 6-month study period, 114 valid CRP-POCTs were performed in the OEMS practice by 18 physicians and the questionnaire was answered in 112 cases (response rate: 98.2%). CRP-POCTs were used in the diagnosis of inflammatory diseases of the gastrointestinal tract (60.0%), respiratory tract infections (17.0%), urinary tract infections (9.0%) and other non-gastrointestinal/non-specified infections (11.0%). The use of a CRP-POCT resulted in a change of the physicians' clinical decision in 83.3% of the cases. Specifically, in 13.6% and 35.1% of the cases, rapid CRP measurements led to decision changes in the (1) initiation of antimicrobial therapy and (2) other drug treatment, respectively. Notably, in 60% of all cases, the use of a CRP-POCT reportedly changed the decision on hospitalisation/non-hospitalisation of OEMS patients. In respect of antimicrobial therapy and hospitalisation, these decision changes primarily (≥73%) promoted 'step-down' decisions, that is, no antibiotic therapy and no hospital admission. In the great majority of CRP-POCT applications (≥95%), OEMS physicians reported that rapid CRP measurements increased the confidence in their diagnostic and therapeutic decision. In almost all cases (97%), physicians rated the CRP-POCT use as useful in the treatment situation., Conclusion: Quantitative CRP-POCT promotes step-down clinical decisions and strengthens the clinical confidence of physicians in out-of-hours OEMS., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

BackgroundAntimicrobial resistance (AMR) is of public health concern worldwide.AimWe aimed to summarise the German AMR situation for clinicians and microbiologists.MethodsWe conducted a systematic review and meta-analysis of 60 published studies and data from the German Antibiotic-Resistance-Surveillance (ARS). Primary outcomes were AMR proportions in bacterial isolates from infected patients in Germany (2016-2021) and the case fatality rates (2010-2021). Random and fixed (common) effect models were used to calculate pooled proportions and pooled case fatality odds ratios, respectively.ResultsThe pooled proportion of meticillin resistance in Staphylococcus aureus infections (MRSA) was 7.9% with a declining trend between 2014 and 2020 (odds ratio (OR) = 0.89; 95% CI: 0.886-0.891; p < 0.0001), while vancomycin resistance in Enterococcus faecium (VRE) bloodstream infections increased (OR = 1.18; (95% CI: 1.16-1.21); p < 0.0001) with a pooled proportion of 34.9%. Case fatality rates for MRSA and VRE were higher than for their susceptible strains (OR = 2.29; 95% CI: 1.91-2.75 and 1.69; 95% CI: 1.22-2.33, respectively). Carbapenem resistance in Gram-negative pathogens ( Klebsiella pneumoniae , Acinetobacter baumannii , Enterobacter spp. and Escherichia coli ) was low to moderate (< 9%), but resistance against third-generation cephalosporins and fluoroquinolones was moderate to high (5-25%). Pseudomonas aeruginosa exhibited high resistance against carbapenems (17.0%; 95% CI: 11.9-22.8), third-generation cephalosporins (10.1%; 95% CI: 6.6-14.2) and fluoroquinolones (24.9%; 95% CI: 19.3-30.9). Statistical heterogeneity was high (I2 > 70%) across studies reporting resistance proportions.ConclusionContinuous efforts in AMR surveillance and infection prevention and control as well as antibiotic stewardship are needed to limit the spread of AMR in Germany.

Background: Due to their fast turnaround time and user-friendliness, point-of-care tests (POCTs) possess a great potential in primary care. The purpose of the study was to assess general practitioners' (GPs) perspectives on POCT use in German primary care, including utilization, limitations and requirements., Methods: We conducted a cross-sectional survey study among GPs in Germany (federal states of Thuringia, Bremen and Bavaria (Lower Franconia), study period: 04/22-06/2022)., Results: From 2,014 GPs reached, 292 participated in our study (response rate: 14.5%). The median number of POCTs used per GP was 7.0 (IQR: 5.0-8.0). Six POCTs are used by the majority of surveyed GPs (> 50%): urine dipstick tests (99%), glucose (urine [91%] and plasma [69%]), SARS-CoV-2 (80%), urine microalbumin (77%), troponin I/T (74%) and prothrombin time / international normalized ratio (65%). The number of utilized POCTs did not differ between GP practice type (p = 0.307) and population size of GP practice location (p = 0.099). The great majority of participating German GPs (93%) rated POCTs as useful diagnostic tools in the GP practice. GPs ranked immediate decisions on patient management and the increase in diagnostic certainty as the most important reasons for performing POCTs. The most frequently reported limitations of POCT use in the GP practice were economic aspects (high costs and inadequate reimbursement), concerns regarding diagnostic accuracy, and difficulties to integrate POCT-testing into practice routines (e.g. time and personnel expenses)., Conclusion: Although participating German GPs generally perceive POCTs as useful diagnostic tools and numerous POCTs are available, several test-related and contextual factors contribute to the relatively low utilization of POCTs in primary care., (© 2023. The Author(s).)

Antimicrobial resistance (AMR) and hospital-acquired infections (HAIs) are global health challenges. The burden of antibiotic resistance in HAIs is still unclear in low- and lower-middle-income countries (L-LMICs). This study summarizes recent data on antibiotic resistance in priority HAIs (ESKAPE-E) in L-LMICs and compares them with data from high-income countries (HICs). EMBASE, Web of Science, and Global Index Medicus were searched for studies on AMR patterns in HAIs published from 01/2010 to 10/2020. Random-effects meta-analyses were performed to obtain pooled estimates. In total, 163 eligible studies were included in the review and meta-analysis. The pooled methicillin resistance proportion in Staphylococcus aureus was 48.4% (95% confidence interval [95%CI] 41·7-55·2, n = 80). Pooled carbapenem resistance proportions were high in Gram-negative pathogens: Escherichia coli : 16·6% (95%CI 10·7-23·4, n = 60); Klebsiella pneumoniae : 34·9% (95%CI 24·6-45·9, n = 50); Pseudomonas aeruginosa : 37.1% (95%CI 24·6-45·9, n = 56); Enterobacter spp.: 51·2% (95%CI 27·5-74·7, n = 7); and Acinetobacter baumannii (complex): 72·4% (95%CI 62·1-81·7%, n = 36). A higher resistance proportions were observed for third-generation cephalosporins: Klebsiella pneumoniae : 78·7% (95%CI 71·5-85·2, n = 46); Escherichia coli: 78·5% (95%CI 72·1-84·2%, n = 58); and Enterobacter spp.: 83·5% (95%CI 71·9-92·8, n = 8). We observed a high between-study heterogeneity (I 2  >  80%), which could not be explained by our set of moderators. Pooled resistance proportions for Gram-negative pathogens were higher in L-LMICs than regional and national estimates from HICs. Patients in resource-constrained regions are particularly affected by AMR. To combat the high resistance to critical antibiotics in L-LMICs, and bridge disparities in health, it is crucial to strengthen local surveillance and the health systems in general.

Introduction: Given their high diagnostic accuracy and fast turnaround time, rapid SARS-CoV-2 tests based on nucleic acid amplification technologies (NAAT) have great potential to expand access to testing and decrease delays in diagnosis of COVID-19., Objectives: The aim of this study was to investigate feasibility, acceptance, organizational consequences and other implementation aspects of the use of a NAAT-based SARS-CoV-2 rapid test (ID NOW™ COVID-19, Abbott Diagnostics) for symptomatic primary care patients with a suspected SARS-CoV-2 infection., Methods: Cross-sectional survey among primary care physicians and medical assistants from Thuringia (Germany) during the third wave of the COVID-19 pandemic in March and April 2021. The health care providers surveyed had previously used the NAAT-based SARS-CoV-2 rapid test as part of a pilot study., Results: Eleven physicians (ten general practitioners and one paediatrician) and 22 medical assistants from Thuringia (Germany) participated in the written survey. Four physicians were additionally interviewed. The majority of the surveyed health care providers rated user-friendliness, integration into practice routine, impact on communication with patients and technical reliability of the NAAT-based SARS-CoV-2 rapid test as (very) positive. Greater workload and the costs for measuring devices were identified as disadvantages compared to PCR laboratory tests. Four out of ten physicians rated the lower sample turnover as unfavourable., Conclusion: Our survey shows that NAAT-based point-of-care SARS-CoV-2 testing gained widespread acceptance among physicians and medical assistants, positively influences workflows, can improve patient communication and could therefore be successfully implemented into routine primary care., (Copyright © 2022. Published by Elsevier GmbH.)

Background: The fast turnaround time and user-friendliness of point-of-care tests (POCTs) offer a great potential to improve outpatient health care where clinical decisions have to be made during the physician-patient encounter and time resources are limited. The aim of this scoping review is to describe the extent and nature as well as gaps in German research activities on POCT in outpatient care. In addition, we define research endpoints that should be addressed in the comprehensive evaluation of POCTs targeted for outpatient care., Methods: We performed a scoping review with a systematic literature search in Medline (via PubMed), Scopus, Web of Science, Cochrane library and Google Scholar for German publications on POCT with relevance to German outpatient care published from January 2005 to November 2020., Results: Our literature search identified 2,200 unique records. After literature selection 117 articles were included in this scoping review. Just over half of the articles (67/117, 57.3%) were primary research studies with original data, while one third of all the studies (33.3%) were secondary research articles (e.g., review articles). The remaining articles were clinical recommendations / position papers (7/117, 6.0%) and other types of articles (3.4%). The majority of articles focused on POCT use in infectious diseases (44/117, 37.6%), diabetic syndromes (15.4%), cardiac disease (12.0%) or coagulopathies and thrombosis (10.3%), while the remaining articles did not specify the disease (13.7%) or investigated other diseases (11.1%). Similar to international studies, most primary research studies investigated the diagnostic performance of POCT (e.g., sensitivity, specificity). Evidence beyond diagnostic accuracy remains scarce, such as the impact on therapeutic decisions and practice routines, clinical effectiveness, and user perspectives. In line with this, interventional studies (such as RCTs) on the effectiveness of POCT use in German outpatient care are limited. We define six endpoint domains that should be addressed in the evaluation of POCTs targeted for outpatient care: (i) diagnostic performance, (ii) clinical performance, (iii) time and costs, (iv) impact on clinical routines / processes, (v) perspectives of medical professionals and patients, and (vi) broader aspects., Conclusion: There is considerable research activity on POCTs targeted for use in outpatient care in Germany. Data on their potential benefits beyond diagnostic accuracy is often lacking and should be addressed in future POCT research studies., (Copyright © 2022. Published by Elsevier GmbH.)

Background: The use of point-of-care tests (POCTs) has been a central strategy to cope with the COVID-19 pandemic. Yet, evidence on the application and consequences of POCTs within medical settings is rare., Purpose: To assess and understand patient perspectives on molecular point-of-care SARS-CoV-2 testing conducted in primary care., Methods: We conducted a cross-sectional survey study among patients who were tested with a molecular SARS-CoV-2 rapid test (ID NOW TM COVID-19 rapid test, Abbott) in 13 primary care practices in the state of Thuringia (Germany) from February to April 2021. The following aspects were covered in the questionnaire through rating scales and open text formats: test characteristics, trust in test result, consequences of immediate result, cost amount willing to pay and expectations in the future. Open text answers were categorized; quantitative data were analyzed using descriptive statistics and a Mann-Whitney U -test to reveal differences in cost contribution depending on the test result., Results: A total of 215 patients from nine family practices and one pediatric practice participated. The immediate availability of the test result was important to the majority of patients (94.3%). 95.7% of patients trusted in their test result. Personal consequences of the immediate test result referred to pandemic measures, certainty of action and reassurance. For further tests, patients were willing to pay between 0€ and 100€ (interquartile range = 10-25€) for the molecular SARS-CoV-2 POCT, regardless of the test result. Expectations of being offered the test again in case of renewed cold symptoms were reported by 96.2%., Conclusion: Patients highly appreciated molecular SARS-CoV-2 rapid testing conducted in primary care practices. The immediate availability of the test result led to adjustments in patients' behavior and emotional wellbeing. However, potentially challenging for the implementation of POCTs in primary care practices may be the reimbursement of test costs and patients' expectations in future situation., Competing Interests: Ms Anni Matthes reports grants from the Ministry of Education and Research, during the conduct of the study. The authors report no other conflicts of interest in this work., (© 2022 Matthes et al.)

BackgroundHospital-acquired infections (HAI) caused by Enterococcus spp., especially vancomycin-resistant Enterococcus spp. (VRE), are of rising concern.AimWe summarised data on incidence, mortality and proportion of HAI caused by enterococci in the World Health Organization European Region.MethodsWe searched Medline and Embase for articles published between 1 January 2010 and 4 February 2020. Random-effects meta-analyses were performed to obtain pooled estimates.ResultsWe included 75 studies. Enterococcus spp. and VRE accounted for 10.9% (95% confidence interval (CI): 8.7-13.4; range: 6.1-17.5) and 1.1% (95% CI: 0.21-2.7; range: 0.39-2.0) of all pathogens isolated from patients with HAI. Hospital wide, the pooled incidence of HAI caused by Enterococcus spp. ranged between 0.7 and 24.8 cases per 1,000 patients (pooled estimate: 6.9; 95% CI: 0.76-19.0). In intensive care units (ICU), pooled incidence of HAI caused by Enterococcus spp. and VRE was 9.6 (95% CI: 6.3-13.5; range: 0.39-36.0) and 2.6 (95% CI: 0.53-5.8; range: 0-9.7). Hospital wide, the pooled vancomycin resistance proportion among Enterococcus spp. HAI isolates was 7.3% (95% CI: 1.5-16.3; range: 2.6-11.5). In ICU, this proportion was 11.5% (95% CI: 4.7-20.1; range: 0-40.0). Among patients with hospital-acquired bloodstream infections with Enterococcus spp., pooled all-cause mortality was 21.9% (95% CI: 15.7-28.9; range: 14.3-32.3); whereas all-cause mortality attributable to VRE was 33.5% (95% CI: 13.0-57.3; range: 14.3-41.3).ConclusionsInfections caused by Enterococcus spp. are frequently identified among hospital patients and associated with high mortality.

The COVID-19 pandemic in Germany has demanded a substantially larger public health workforce to perform contact tracing and contact management of COVID-19 cases, in line with recommendations of the World Health Organization (WHO). In response, the Robert Koch Institute (RKI) established the national "Containment Scout Initiative" (CSI) to support the local health authorities with a short-term workforce solution. It is part of a range of measures for strengthening the public health system in order to limit the spread of SARS-CoV-2 in Germany. The CSI is an example of how solutions to address critical health system capacity issues can be implemented quickly. It also demonstrates that medical or health-related backgrounds may not be necessary to support health authorities with pandemic-specific tasks and fulfil accurate contact tracing. However, it is a short-term solution and cannot compensate for the lack of existing qualified staff as well as other deficits that exist within the public health sector in Germany. This article describes the structure and process of the first phase of this initiative in order to support health policymakers, public health practitioners, and researchers considering innovative and flexible approaches for addressing urgent workforce capacity issues.

Background: Neonates are at major risk of sepsis, but data on neonatal sepsis incidence are scarce. We aimed to assess the incidence and mortality of neonatal sepsis worldwide., Methods: We performed a systematic review and meta-analysis. 13 databases were searched for the period January 1979-May 2019, updating the search of a previous systematic review and extending it in order to increase data inputs from low-income and middle-income countries (LMICs). We included studies on the population-level neonatal sepsis incidence that used a clinical sepsis definition, such as the 2005 consensus definition, or relevant ICD codes. We performed a random-effects meta-analysis on neonatal sepsis incidence and mortality, stratified according to sepsis onset, birth weight, prematurity, study setting, WHO region and World Bank income level., Results: The search yielded 4737 publications, of which 26 were included. They accounted for 2 797 879 live births and 29 608 sepsis cases in 14 countries, most of which were middle-income countries. Random-effects estimator for neonatal sepsis incidence in the overall time frame was 2824 (95% CI 1892 to 4194) cases per 100 000 live births, of which an estimated 17.6% 9 (95% CI 10.3% to 28.6%) died. In the last decade (2009-2018), the incidence was 3930 (95% CI 1937 to 7812) per 100 000 live births based on four studies from LMICs. In the overall time frame, estimated incidence and mortality was higher in early-onset than late-onset neonatal sepsis cases. There was substantial between-study heterogeneity in all analyses. Studies were at moderate to high risk of bias., Conclusion: Neonatal sepsis is common and often fatal. Its incidence remains unknown in most countries and existing studies show marked heterogeneity, indicating the need to increase the number of epidemiological studies, harmonise neonatal sepsis definitions and improve the quality of research in this field. This can help to design and implement targeted interventions, which are urgently needed to reduce the high incidence of neonatal sepsis worldwide., Competing Interests: Competing interests: CF received grants from the German Federal Ministry of Education and Research (BMBF) via the Center for Sepsis Control and Care (FKZ: 01EO1002 and 01EO1502) and from the Innovation Funds of the German Government (FKZ 01VSF17010). The other authors declared no competing interests., (© Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY. Published by BMJ.)

Data on microbiological profiles in odontogenic infections are scarce. This study aimed to analyze the spectrum of pathogens and antimicrobial resistance in clinical isolates from dental and oral-maxillofacial clinical settings in Germany. We analyzed 20,645 clinical isolates (dental practices: n = 5,733; hospitals: n = 14,912) from patients with odontogenic infections using data (2012-2019) from the German Antimicrobial-Resistance-Surveillance (ARS) system. A total of 224 different species from 73 genera were found in clinical isolates from dental practices, and 329 different species from 97 genera were identified in isolates from hospital patients. In both hospitals and dental practices Streptococcus spp. (33 and 36%, respectively) and Staphylococcus spp. (21 and 12%, respectively) were the most frequently isolated microorganisms. In Streptococcus spp. isolates from hospitals, penicillin and aminopenicillin resistance proportions were 8.0% (95%CI 4.7-14.9%) and 6.9% (95%CI 4.7-9.9%), respectively. Substantially lower resistance proportions of penicillin and aminopenicillin were observed in dental practices [2.6% (95%CI 1.4-4.7%) and 2.1% (95%CI 1.1-4.0%), respectively]. Among Staphylococcus aureus isolates from hospital patients methicillin resistance proportions were 12.0% (95%CI 9.7-14.8%), which was higher than in isolates from dental practices (5.8% (95%CI 4.1-8.1%)]. High clindamycin and macrolide resistance proportions (>17%) were observed in Streptococcus spp. and Staphylococcus aureus isolates. In Klebsiella spp. isolates carbapenem resistance proportions were <1%. In sum, substantial antibiotic resistance was observed in isolates from odontogenic infections, which calls for strengthened efforts in antibiotic stewardship and infection prevention and control measures in both hospitals and dental practices., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Meinen, Reuss, Willrich, Feig, Noll, Eckmanns, Al-Nawas and Markwart.)

Vancomycin-resistant Enterococcus faecium (VREF) and methicillin-resistant Staphylococcus aureus (MRSA) are associated with significant health burden. We investigated linezolid and daptomycin resistance among VREF and MRSA in the EU/EEA between 2014 and 2018. Descriptive statistics and multivariable logistic regression were used to analyze 6,949 VREF and 35,131 MRSA blood isolates from patients with bloodstream infection. The population-weighted mean proportion of linezolid resistance in VREF and MRSA between 2014 and 2018 was 1.6% (95% CI 1.33-2.03%) and 0.28% (95% CI 0.32-0.38%), respectively. Daptomycin resistance in MRSA isolates was similarly low [1.1% (95% CI 0.75-1.6%)]. On the European level, there was no temporal change of daptomycin and linezolid resistance in MRSA and VREF. Multivariable regression analyses showed that there was a higher likelihood of linezolid and daptomycin resistance in MRSA (aOR: 2.74, p < 0.001; aOR: 2.25, p < 0.001) and linezolid in VREF (aOR: 1.99, p < 0.001) compared to their sensitive isolates. The low proportion of linezolid and daptomycin resistance in VREF and MRSA suggests that these last-resort antibiotics remain effective and will continue to play an important role in the clinical management of these infections in Europe. However, regional and national efforts to contain antimicrobial resistance should continue to monitor the trend through strengthened surveillance that includes genomic surveillance for early warning and action., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Markwart, Willrich, Eckmanns, Werner and Ayobami.)

Background: Carbapenem-resistant Acinetobacter baumannii complex (CRABC) has globally emerged as a serious public health challenge. This study aimed to describe epidemiological trends and risk factors of carbapenem resistance in A. baumannii complex isolates in Germany between 2014 and 2018., Methods: We analysed 43,948 clinical A. baumannii complex isolates using 2014 to 2018 data from the German Antimicrobial Resistance Surveillance system. We applied descriptive statistics and uni- and multivariable regression analyses to investigate carbapenem resistance in A. baumannii complex isolates., Results: The proportion of carbapenem resistance in clinical A. baumannii complex isolates declined from 7.6% (95% confidence interval [95% CI] 4.4-12.7%) in 2014 to 3.5% (95% CI 2.5-4.7%) in 2018 (adjusted OR [aOR] 0.85 [95% CI 0.79-0.93, p ≤ 0.001]). Higher mean CRABC proportions for 2014 to 2018 were observed in secondary care hospitals (4.9% [95% CI 3.2-7.5%], aOR 3.6 [95% CI 2.4-5.3, p ≤ 0.001]) and tertiary care hospitals (5.9% [95% CI 3.0-11.2%], aOR 5.4 [95% CI 2.9-10.0, p ≤ 0.001) compared to outpatient clinics (1.3% [95% CI 1.1-1.6%]). CRABC proportions in hospitals varied between German regions and ranged between 2.4% (95% CI 1.6-3.5%) in the Southeast and 8.8% (95% CI 4.2-17.3%) in the Northwest. Lower CRABC proportions were observed in younger patients (< 1 year: 0.6% [95% CI 0.2-1.3%]; 1-19 years: 1.3% [95% CI 0.7-2.5%]) than adults (20-39 years: 7.7% [95% CI 4.4-13.0%]; 40-59 years: 6.2% [4.2-8.9%]; 60-79 years: 5.8% [95% CI 4.0-8.3%]). In the 20-39 year old patient age group, CRABC proportions were significantly higher for men than for women (14.6% [95% CI 8.6-23.6%] vs. 2.5% [95% CI 1.3-4.5%]). A. baumannii complex isolates from lower respiratory infections were more likely to be carbapenem-resistant than isolates from upper respiratory infections (11.4% [95% CI 7.9-16.2%] vs. 4.0% [95% CI 2.7-6.0%]; adjusted OR: 1.5 [95% CI 1.2-1.9, p ≤ 0.001])., Conclusions: In contrast to many other regions worldwide, carbapenem resistance proportions among clinical A. baumannii complex isolates are relatively low in Germany and have declined in the last few years. Ongoing efforts in antibiotic stewardship and infection prevention and control are needed to prevent the spread of carbapenem-resistant A. baumannii complex in Germany.

Vancomycin-resistant enterococci infections are of great public health significance due to limited therapeutic options. We investigated epidemiological trends and risk factors of vancomycin resistance in enterococci isolates from patients with bloodstream infections in the EU/EEA from 2012 to 2018. Routine vancomycin susceptibility data of clinical E. faecium (n = 67,022) and E. faecalis (n = 103,112) blood isolates from the European Antimicrobial Resistance Surveillance Network were analysed using descriptive statistics and multivariable regression analyses. In Europe, proportions of vancomycin-resistant E. faecium (VREFm) increased from 8.1% (95%CI 6.7-9.7%) in 2012 to 19.0% (95%CI 16.8-21.5%) in 2018. Rising VREFm proportions were observed across all European regions, both genders and all age groups except children and adolescents (1-19 years). Adults (20-59 years) and elderly (≥60 years) had an increased likelihood of VREFm compared to children and adolescents (1-19 years) (OR: 1.99 [95%CI 1.42-2.79, p < 0.001] and OR: 1.56 [95%CI 1.09-2.23, p = 0.014], respectively). Inpatients hospital units, including internal medicine and ICUs, were associated with an increased likelihood of VREFm (OR: 2.29 (95%CI 1.58-3.32, p < 0.001) compared to the emergency department which reflects patients with community origin of E. faecium infections. The mean proportion of vancomycin-resistant E. faecalis in Europe was found to be low (1.1% [95%CI 0.9-1.4%]). Local and regional authorities should intensify efforts directed at diagnostic and antimicrobial stewardship for vancomycin and all last resort drugs for the management of VREFm, particularly for hospitalized elderly patients.

Purpose: Sepsis is recognized as a global public health problem, but the proportion due to hospital-acquired infections remains unclear. We aimed to summarize the epidemiological evidence related to the burden of hospital-acquired (HA) and ICU-acquired (ICU-A) sepsis., Methods: We searched MEDLINE, Embase and the Global Index Medicus from 01/2000 to 03/2018. We included studies conducted hospital-wide or in intensive care units (ICUs), including neonatal units (NICUs), with data on the incidence/prevalence of HA and ICU-A sepsis and the proportion of community and hospital/ICU origin. We did random-effects meta-analyses to obtain pooled estimates; inter-study heterogeneity and risk of bias were assessed., Results: Of the 13,239 studies identified, 51 met the inclusion criteria; 22 were from low- and middle-income countries. Twenty-eight studies were conducted in ICUs, 13 in NICUs, and ten hospital-wide. The proportion of HA sepsis among all hospital-treated sepsis cases was 23.6% (95% CI 17-31.8%, range 16-36.4%). In the ICU, 24.4% (95% CI 16.7-34.2%, range 10.3-42.5%) of cases of sepsis with organ dysfunction were acquired during ICU stay and 48.7% (95% CI 38.3-59.3%, range 18.7-69.4%) had a hospital origin. The pooled hospital incidence of HA sepsis with organ dysfunction per 1000 patients was 9.3 (95% CI 7.3-11.9, range 2-20.6)). In the ICU, the pooled incidence of HA sepsis with organ dysfunction per 1000 patients was 56.5 (95% CI 35-90.2, range 9.2-254.4) and it was particularly high in NICUs. Mortality of ICU patients with HA sepsis with organ dysfunction was 52.3% (95% CI 43.4-61.1%, range 30.1-64.6%). There was a significant inter-study heterogeneity. Risk of bias was low to moderate in ICU-based studies and moderate to high in hospital-wide and NICU studies., Conclusion: HA sepsis is of major public health importance, and the burden is particularly high in ICUs. There is an urgent need to improve the implementation of global and local infection prevention and management strategies to reduce its high burden among hospitalized patients.

Background: Due to limited therapeutic options and their association with high mortality and morbidity, carbapenem-non-susceptible Acinetobacter spp. (CNA) are of significant public health importance. This study aimed to describe current epidemiological trends of CNA proportions in Europe and to identify factors that are associated with carbapenem non-susceptibility of isolates from patients with invasive Acinetobacter spp. infections., Methods: Data from routine carbapenem susceptibility testing of 18,412 invasive clinical Acinetobacter spp. isolates from 30 European countries in 2013-2017 were analysed using descriptive statistical analyses and uni- and multivariable regression analyses. These data were obtained from the European Antimicrobial Resistance Surveillance Network (EARS-Net)., Results: The population-weighted mean proportion of carbapenem-non-susceptible Acinetobacter spp. in Europe is 35.6% (95% confidence interval [CI] 29.7-42.0%). With CNA proportions of 75.5% (95% CI 71.2-79.4%) and 71.5% (95% CI 66.7-75.9%) the burden of CNA is particularly high in Southern and Eastern European regions. In contrast, Northern and Western European regions recorded CNA proportions of 2.8% (95% CI 1.2-6.0%) and 6.3% (95% CI 4.5-8.9%), respectively. Population-weighted mean CNA proportions are especially high in Acinetobacter spp. isolates from intensive care units (54.0% [95% CI 47.6-60.3%]). Male gender, age above 20 years and ICU admission were identified as independent factors associated with an increased likelihood of CNA., Conclusion: The burden of carbapenem-non-susceptible Acinetobacter spp. is particularly high in Southern and Eastern Europe. There is a risk that resistance could spread to other parts of Europe. Therefore, increased efforts in infection control and antibiotic stewardship, particularly in Intensive Care Units, are necessary to combat the spread of CNA in Europe.

Background: Due to limited therapeutic options, vancomycin-resistant Enterococcus faecium (VREF) is of great clinical significance. Recently, rising proportions of vancomycin resistance in enterococcal infections have been reported worldwide. This study aims to describe current epidemiological trends of VREF in German hospitals and to identify factors that are associated with an increased likelihood of vancomycin resistance in clinical E. faecium isolates., Methods: 2012 to 2017 data from routine vancomycin susceptibility testing of 35,906 clinical E. faecium isolates from 148 hospitals were analysed using data from the German Antimicrobial Resistance Surveillance System. Descriptive statistical analyses and uni- and multivariable regression analyses were performed to investigate the impact of variables, such as year of sampling, age and region, on vancomycin resistance in clinical E. faecium isolates., Results: From 2014 onwards the proportions of clinical E. faecium isolates exhibiting resistance to vancomycin increased from 11.2% (95% confidence interval [CI] 9.4-13.3%) to 26.1% (95% CI 23.1-29.4%) in 2017. The rise of VREF proportions is primarily observed in the southern regions of Germany, whereas northern regions do not show a major increase. In the Southwest and Southeast, VREF proportions increased from 10.8% (95% CI 6.9-16.5%) and 3.8% (95% CI 3.0-11.5%) in 2014 to 36.7% (95% CI 32.9-40.8%) and 36.8% (95% CI 29.2-44.7%) in 2017, respectively. VREF proportions were considerably higher in isolates from patients aged 40-59 years compared to younger patients. Further regression analyses show that in relation to secondary care hospitals, E. faecium samples collected in specialist care hospitals and prevention and rehabilitation care centres are more likely to be vancomycin-resistant (odds ratios: 2.4 [95% CI 1.2-4.6] and 2.4 [95% CI 1.9-3.0], respectively). No differences in VREF proportions were found between female and male patients as well as between different clinical specimens., Conclusion: The proportion of VREF is increasing in German hospitals, particularly in southern regions in Germany. Increased efforts in infection control and antibiotic stewardship activities accounting for local resistance patterns are necessary to combat the spread of VREF in Germany., Competing Interests: Competing interestsThe authors declare that they have no competing interests.

T lymphocytes (T cells) are highly specialized members of the adaptive immune system and hold the key to the understanding the hosts' response toward invading pathogen or pathogen-associated molecular patterns such as LPS. In this study, noninvasive Raman spectroscopy is presented as a label-free method to follow LPS-induced changes in splenic T cells during acute and postacute inflammatory phases (1, 4, 10, and 30 d) with a special focus on CD4 + and CD8 + T cells of endotoxemic C57BL/6 mice. Raman spectral analysis reveals highest chemical differences between CD4 + and CD8 + T cells originating from the control and LPS-treated mice during acute inflammation, and the differences are visible up to 10 d after the LPS insult. In the postacute phase, CD4 + and CD8 + T cells from treated and untreated mice could not be differentiated anymore, suggesting that T cells largely regained their original status. In sum, the biological information obtained from Raman spectra agrees with immunological readouts demonstrating that Raman spectroscopy is a well-suited, label-free method for following splenic T cell activation in systemic inflammation from acute to postacute phases. The method can also be applied to directly study tissue sections as is demonstrated for spleen tissue one day after LPS insult., (Copyright © 2019 The Authors.)

Due to therapeutic challenges, hospital-acquired infections (HAIs) caused by Acinetobacter baumannii (HA-AB), particularly carbapenem-resistant strains (HA-CRAB) pose a serious health threat to patients worldwide. This systematic review sought to summarize recent data on the incidence and prevalence of HA-AB and HA-CRAB infections in the WHO-defined regions of Europe (EUR), Eastern Mediterranean (EMR) and Africa (AFR). A comprehensive literature search was performed using MEDLINE, EMBASE and GMI databases (01/2014-02/2019). Random-effects meta-analyses were performed to determine the pooled incidence of HA-AB and HA-CRAB infections as well as the proportions of A. baumannii among all HAIs. 24 studies from 3,340 records were included in this review (EUR: 16, EMR: 6, AFR: 2). The pooled estimates of incidence and incidence density of HA-AB infection in intensive care units (ICUs) were 56.5 (95% CI 33.9-92.8) cases per 1,000 patients and 4.4 (95% CI 2.9-6.6) cases per 1,000 patient days, respectively. Five studies conducted at a hospital-wide level or in specialized clinical departments/wards (ICU + non-ICU patients) showed HA-AB incidences between 0.85 and 5.6 cases per 1,000 patients. For carbapenem-resistant A. baumannii infections in ICUs, the pooled incidence and incidence density were 41.7 (95% CI 21.6-78.7) cases per 1,000 patients and 2.1 (95% CI 1.2-3.7) cases per 1,000 patient days, respectively. In ICUs, A. baumannii and carbapenem-resistant A. baumannii strains accounted for 20.9% (95% CI 16.5-26.2%) and 13.6% (95% CI 9.7-18.7%) of all HAIs, respectively. Our study highlights the persistent clinical significance of hospital-acquired A. baumannii infections in the studied WHO regions, particularly in ICUs.

The palatine tonsils, localized in the oropharynx, are easily accessible secondary lymphoid tissue in humans. Inflammation of the palatine tonsils, local and chronic in case of chronic tonsillitis (CT) or acute in the presence of a peritonsillar abscess (PTA), ranks among the most common diseases in otolaryngology. However, the functionality of tonsillar immune cells, notably T-cells, in the context of these immune pathologies is poorly understood. We have examined the functional status of human tonsillar T-cells in CT and compared it to the acute inflammatory setting of a PTA. Patients presenting with CT (n = 10) or unilateral PTA (n = 7) underwent bilateral tonsillectomy and a subgroup of 8 patients underwent additional blood sampling. T-cells were purified via automated magnetic selection and subjected to flow cytometry-based immunophenotyping. In addition, the response to T-cell receptor (TCR) stimulation was assessed at the level of proximal signaling, activation marker expression and proliferation. We observed no difference between the percentage of T helper (CD4(+)) cells from tonsil tissue in CT and PTA, but observed a trend towards a higher percentage of T helper cells in the blood of patients with PTA versus CT, probably reflecting an acute, systemic bacterial infection in the former cohort. Tonsils from CT harbored more PD-1(+) CD4(+) T-cells, pointing to T-cell exhaustion due to chronic infection. This notion was supported by functional studies that showed a tendency to weaker TCR responses of tonsillar T-cells from CT. Intriguingly, tonsillar T-cells recurrently featured a dampened response to T-cell receptor stimulation at the level of receptor proximal signaling steps compared to peripheral T-cells. In sum, our study documents distinct differences in tonsillar T-cell class distribution and function between the various pathological conditions. Our observations are consistent with the concept that tonsillar T-cells react to infections by eliciting specific immunological responses in chronic versus acute settings of inflammation.

Sepsis is characterized by a disproportionate host response to infection that often culminates in multiple organ failure. Current concepts invoke a deregulated immune reaction involving features of hyperinflammation, as well as protracted immune suppression. However, owing to the scarcity of human data, the precise origin of a long-term suppression of adaptive immunity remains doubtful. We report on an explorative clinical study of chronic critical illness (CCI) patients aimed at assessing the long-term consequences of sepsis on T cell function. Blood was drawn from 12 male CCI patients (median age 67 y, range 48-79 y) receiving continuous mechanical ventilation and renal replacement therapy in a long-term care hospital who had been treated in an external acute care hospital for severe sepsis. T cells were purified and subjected to flow cytometric immune-phenotyping and functional assays. We found that T cells from CCI patients featured higher basal levels of activation and stronger expression of the inhibitory surface receptor programmed cell death 1 compared with controls. However, T cells from CCI patients exhibited no suppressed TCR response at the level of proximal TCR signaling (activation/phosphorylation of PLCγ, Erk, Akt, LAT), activation marker upregulation (CD69, CD25, CD154, NUR77), IL-2 production, or clonal expansion. Rather, our data illustrate an augmented response in T cells from CCI patients in response to TCR/coreceptor (CD3/CD28) challenge. Thus, the present findings reveal that CCI sepsis patients feature signs of immune suppression but that their T cells exhibit a primed, rather than a suppressed, phenotype in their TCR response, arguing against a generalized T cell paralysis as a major cause of protracted immune suppression from sepsis., (Copyright © 2017 by The American Association of Immunologists, Inc.)

Hepatic stellate cells (HSCs) are retinoid storing cells in the liver: The retinoid content of those cells changes depending on nutrition and stress level. There are also differences with regard to a HSC's anatomical position in the liver. Up to now, retinoid levels were only accessible from bulk measurements of tissue homogenates or cell extracts. Unfortunately, they do not account for the intercellular variability. Herein, Raman spectroscopy relying on excitation by the minimally destructive wavelength 785 nm is introduced for the assessment of the retinoid state of single HSCs in freshly isolated, unprocessed murine liver lobes. A quantitative estimation of the cellular retinoid content is derived. Implications of the retinoid content on hepatic health state are reported. The Raman-based results are integrated with histological assessments of the tissue samples. This spectroscopic approach enables single cell analysis regarding an important cellular feature in unharmed tissue.

Background: Growth factors induce a characteristically short-lived Ras activation in cells emerging from quiescence. Extensive work has shown that transient as opposed to sustained Ras activation is critical for the induction of mitogenic programs. Mitogen-induced accumulation of active Ras-GTP results from increased nucleotide exchange driven by the nucleotide exchange factor Sos. In contrast, the mechanism accounting for signal termination and prompt restoration of basal Ras-GTP levels is unclear, but has been inferred to involve feedback inhibition of Sos. Remarkably, how GTP-hydrolase activating proteins (GAPs) participate in controlling the rise and fall of Ras-GTP levels is unknown., Results: Monitoring nucleotide exchange of Ras in permeabilized cells we find, unexpectedly, that the decline of growth factor-induced Ras-GTP levels proceeds in the presence of unabated high nucleotide exchange, pointing to GAP activation as a major mechanism of signal termination. Experiments with non-hydrolysable GTP analogues and mathematical modeling confirmed and rationalized the presence of high GAP activity as Ras-GTP levels decline in a background of high nucleotide exchange. Using pharmacological and genetic approaches we document a raised activity of the neurofibromatosis type I tumor suppressor Ras-GAP neurofibromin and an involvement of Rsk1 and Rsk2 in the down-regulation of Ras-GTP levels., Conclusions: Our findings show that, in addition to feedback inhibition of Sos, feedback stimulation of the RasGAP neurofibromin enforces termination of the Ras signal in the context of growth-factor signaling. These findings ascribe a precise role to neurofibromin in growth factor-dependent control of Ras activity and illustrate how, by engaging Ras-GAP activity, mitogen-challenged cells play safe to ensure a timely termination of the Ras signal irrespectively of the reigning rate of nucleotide exchange.

Increasing clinical evidence indicates that removal of the palatine tonsils enhances the risk for adults to suffer from severe illnesses. Together with recent experimental findings pointing to the presence of immunologically competent immune cells these findings illustrate that adult palatine tonsils likely play an appreciable role in the host immune response. T-cells are abundant in the palatine tonsil and are a pivotal entity of the adaptive immune response. However, investigation of T-cells from tonsils has been widely neglected and largely restricted to immune phenotyping. Accordingly, methodological literature describing the experimental preparation and isolation of T-cells from tonsils is scarce and has rarely been complemented with rigorous tests of T-cell functionality. We report here on a comparative investigation of three isolation protocols composed of permutations of different tissue grinding approaches, density gradient centrifugation and automated magnetic collection of CD4/CD8 T-cells. Importantly we put a strong emphasis on assessing the impact of the preparative procedures on the functionality of T-cells at the level of viability and functional response to T-cell receptor (TCR) ligation. The reported, optimized preparation protocols allow for the rapid isolation of highly viable, functional T-cells within 2.5h and represent a useful, affordable approach for the analysis of tonsillar T-cells., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Ras is a prototypical small G-protein and a central regulator of growth, proliferation and differentiation processes in virtually every nucleated cell. As such, Ras becomes engaged and activated by multiple growth factors, mitogens, cytokines or adhesion receptors. Ras activation comes about by changes in the steady-state equilibrium between the inactive guanosine diphosphate (GDP)-bound and active guanosine triphosphate (GTP)-bound states of Ras, resulting in the mostly transient accumulation of Ras-GTP. Three decades of intense Ras research have disclosed various families of guanine nucleotide exchange factors (GEFs) and GTPase activating proteins (GAPs) as the two principal regulatory elements of the Ras-GDP/GTP loading status. However, with the possible exception of the GEF Sos, we still have only a rudimentary knowledge of the precise role played by many GEF and GAP members in the signalling network upstream of Ras. As for GAPs, we even lack the fundamental understanding of whether they function as genuine signal transducers in the context of growth factor-elicited Ras activation or rather act as passive modulators of the Ras-GDP/GTP cycle. Here we sift through the large body of Ras literature and review the relevant data for understanding the participation and precise role played by GEFs and GAPs in the process of Ras activation.

Sepsis describes the life-threatening systemic inflammatory response (SIRS) of an organism to an infection and is the leading cause of mortality on intensive care units (ICU) worldwide. An acute episode of sepsis is characterized by the extensive release of cytokines and other mediators resulting in a dysregulated immune response leading to organ damage and/or death. This initial pro-inflammatory burst often transits into a state of immune suppression characterised by loss of immune cells and T-cell dysfunction at later disease stages in sepsis survivors. However, despite these appreciations, the precise nature of the evoked defect in T-cell immunity in post-acute phases of SIRS remains unknown. Here we present an in-depth functional analysis of T-cell function in post-acute SIRS/sepsis. We document that T-cell function is not compromised on a per cell basis in experimental rodent models of infection-free SIRS (LPS or CpG) or septic peritonitis. Transgenic antigen-specific T-cells feature an unaltered cytokine response if challenged in vivo and ex vivo with cognate antigens. Isolated CD4(+)/CD8(+) T-cells from post-acute septic animals do not exhibit defects in T-cell receptor-mediated activation at the the level of receptor-proximal signalling, activation marker upregulation or expansion. However, SIRS/sepsis induced transient lymphopenia and gave rise to an environment of immune attenuation at post acute disease stages. Thus, systemic inflammation has an acute impact on T-cell numbers and adaptive immunity, but does not cause major cell-autonomous enduring functional defects in T-cells.

Receptor tyrosine kinases participate in several signaling pathways through small G proteins such as Ras (rat sarcoma). An important component in the activation of these G proteins is Son of sevenless (SOS), which catalyzes the nucleotide exchange on Ras. For optimal activity, a second Ras molecule acts as an allosteric activator by binding to a second Ras-binding site within SOS. This allosteric Ras-binding site is blocked by autoinhibitory domains of SOS. We have reported recently that Ras activation also requires the actin-binding proteins ezrin, radixin, and moesin. Here we report the mechanism by which ezrin modulates SOS activity and thereby Ras activation. Active ezrin enhances Ras/MAPK signaling and interacts with both SOS and Ras in vivo and in vitro. Moreover, in vitro kinetic assays with recombinant proteins show that ezrin also is important for the activity of SOS itself. Ezrin interacts with GDP-Ras and with the Dbl homology (DH)/pleckstrin homology (PH) domains of SOS, bringing GDP-Ras to the proximity of the allosteric site of SOS. These actions of ezrin are antagonized by the neurofibromatosis type 2 tumor-suppressor protein merlin. We propose an additional essential step in SOS/Ras control that is relevant for human cancer as well as all physiological processes involving Ras.

Ras GTPases undergo post-translational modifications that govern their subcellular trafficking and localization. In particular, palmitoylation of the Golgi tags N-Ras and H-Ras for exocytotic transport and residency at the PM (plasma membrane). Following depalmitoylation, PM-Ras redistributes to all subcellular membranes causing an accumulation of palmitate-free Ras at endomembranes, including the Golgi and endoplasmic reticulum. Palmitoylation is unanimously regarded as a critical modification at the crossroads of Ras activity and trafficking control, but its precise relevance to native wild-type Ras function in growth factor signalling is unknown. We show in the present study by use of palmitoylation-deficient N-Ras mutants and via the analysis of palmitate content of agonist-activated GTP-loaded N-Ras that only palmitoylated N-Ras becomes activated by agonists. In line with an essential role of palmitoylation in Ras activation, dominant-negative RasS17N loses its blocking potency if rendered devoid of palmitoylation. Live-cell Ras-GTP imaging shows that N-Ras activation proceeds only at the PM, consistent with activated N-Ras-GTP being palmitoylated. Finally, palmitoylation-deficient N-Ras does not sustain EGF (epidermal growth factor) or serum-elicited mitogenic signalling, confirming that palmitoylation is essential for signal transduction by N-Ras. These findings document that N-Ras activation proceeds at the PM and suggest that depalmitoylation, by removing Ras from the PM, may contribute to the shutdown of Ras signalling.