The Overlooked Microbiome-Considering Archaea and Eukaryotes Using Multiplex Nanopore-16S-/18S-rDNA-Sequencing: A Technical Report Focusing on Nasopharyngeal Microbiomes.

In contrast to bacteria, microbiome analyses often neglect archaea, but also eukaryotes. This is partly because they are difficult to culture due to their demanding growth requirements, or some even have to be classified as uncultured microorganisms. Consequently, little is known about the relevance of archaea in human health and diseases. Contemporary broad availability and spread of next generation sequencing techniques now enable a stronger focus on such microorganisms, whose cultivation is difficult. However, due to the enormous evolutionary distances between bacteria, archaea and eukaryotes, the implementation of sequencing strategies for smaller laboratory scales needs to be refined to achieve as a holistic view on the microbiome as possible. Here, we present a technical approach that enables simultaneous analyses of archaeal, bacterial and eukaryotic microbial communities to study their roles in development and courses of respiratory disorders. We thus applied combinatorial 16S-/18S-rDNA sequencing strategies for sequencing-library preparation. Considering the lower total microbiota density of airway surfaces, when compared with gut microbiota, we optimized the DNA purification workflow from nasopharyngeal swab specimens. As a result, we provide a protocol that allows the efficient combination of bacterial, archaeal, and eukaryotic libraries for nanopore-sequencing using Oxford Nanopore Technologies MinION devices and subsequent phylogenetic analyses. In a pilot study, this workflow allowed the identification of some environmental archaea, which were not correlated with airway microbial communities before. Moreover, we assessed the protocol's broader applicability using a set of human stool samples. We conclude that the proposed protocol provides a versatile and adaptable tool for combinatorial studies on bacterial, archaeal, and eukaryotic microbiomes on a small laboratory scale.

Do Current Asthma-Preventive Measures Appropriately Face the World Health Organization's Concerns: A Study Presentation of a New Clinical, Prospective, Multicentric Pediatric Asthma Exacerbation Cohort in Germany.

In summer 2017, the World Health Organization published 10 facts on asthma, which is known as a major non-communicable disease of high clinical and scientific importance with currently several hundred million people-with many children among them-suffering from air passages inflammation and narrowing. Importantly, the World Health Organization sees asthma as being underdiagnosed and undertreated. Consequently, much more efforts in clinical disease management and research need to be spent on reducing the asthma-related health burden. Particularly, for young approximately 6 months aged patients presenting recurrent bronchitic respiratory symptoms, many parents anxiously ask the doctors for risk prognosis for their children's future life. Therefore, we urgently need to reevaluate if the current diagnostic and treatment measures are in concordance with our yet incomplete knowledge of pathomechanisms on exacerbation. To contribute to this increasing concern worldwide, we established a multicentric pediatric exacerbation study network, still recruiting acute exacerbated asthmatics (children >6 years) and preschool asthmatics/wheezers (children <6 years) since winter 2018 in Germany. The current study that has a currently population comprising 176 study participants aims to discover novel holistic entry points for achieving a better understanding of the poorly understood plasticity of involved molecular pathways and to define biomarkers enabling improved diagnostics and therapeutics. With this study description, we want to present the study design, population, and few ongoing experiments for novel biomarker research. Clinical Trial Registration: German Clinical Trials Register (Deutsches Register für Klinische Studien, DRKS): DRKS00015738.

Grape or grain but never the twain? A randomized controlled multiarm matched-triplet crossover trial of beer and wine.

Background: Alcohol-induced hangover constitutes a significant, yet understudied, global hazard and a large socio-economic burden. Old folk wisdoms such as "Beer before wine and you'll feel fine; wine before beer and you'll feel queer" exist in many languages. However, whether these concepts in fact reduce hangover severity is unclear. Objectives: The aim of this study was to investigate the influence of the combination and order of beer and wine consumption on hangover intensity. Methods: In this multiarm, parallel randomized controlled matched-triplet crossover open-label interventional trial, participants were matched into triplets and randomly assigned according to age, gender, body composition, alcohol drinking habits, and hangover frequency. Study group 1 consumed beer up to a breath alcohol concentration (BrAC) ≥0.05% and then wine to BrAC ≥0.11% (vice versa for study group 2). Control group subjects consumed either only beer or only wine. On a second intervention day (crossover) ≥1 wk later, study-group subjects were switched to the opposite drinking order. Control-group subjects who drank only beer on the first intervention received only wine on the second study day (and vice versa). Primary endpoint was hangover severity assessed by Acute Hangover Scale rating on the day following each intervention. Secondary endpoints were factors associated with hangover intensity. Results: Ninety participants aged 19-40 y (mean age 23.9), 50% female, were included (study group 1 n = 31, study group 2 n = 31, controls n = 28). Neither type nor order of consumed alcoholic beverages significantly affected hangover intensity (P > 0.05). Multivariate regression analyses revealed perceived drunkenness and vomiting as the strongest predictors for hangover intensity. Conclusions: Our findings dispel the traditional myths "Grape or grain but never the twain" and "Beer before wine and you'll feel fine; wine before beer and you'll feel queer" regarding moderate-to-severe alcohol intoxication, whereas subjective signs of progressive intoxication were confirmed as accurate predictors of hangover severity. This trial was prospectively registered at the Witten/Herdecke University Ethics Committee as 140/2016 and retrospectively registered at the German Clinical Trials Register as DRKS00015285.

Moxifloxacin in Pediatric Patients With Complicated Intra-abdominal Infections: Results of the MOXIPEDIA Randomized Controlled Study.

BACKGROUND: This study was designed to evaluate primarily the safety and also the efficacy of moxifloxacin (MXF) in children with complicated intra-abdominal infections (cIAIs). METHODS: In this multicenter, randomized, double-blind, controlled study, 451 pediatric patients aged 3 months to 17 years with cIAIs were treated with intravenous/oral MXF (N = 301) or comparator (COMP, intravenous ertapenem followed by oral amoxicillin/clavulanate; N = 150) for 5 to 14 days. Doses of MXF were selected based on the results of a Phase 1 study in pediatric patients (NCT01049022). The primary endpoint was safety, with particular focus on cardiac and musculoskeletal safety; clinical and bacteriologic efficacy at test of cure was also investigated. RESULTS: The proportion of patients with adverse events (AEs) was comparable between the 2 treatment arms (MXF: 58.1% and COMP: 54.7%). The incidence of drug-related AEs was higher in the MXF arm than in the COMP arm (14.3% and 6.7%, respectively). No cases of QTc interval prolongation-related morbidity or mortality were observed. The proportion of patients with musculoskeletal AEs was comparable between treatment arms; no drug-related events were reported. Clinical cure rates were 84.6% and 95.5% in the MXF and COMP arms, respectively, in patients with confirmed pathogen(s) at baseline. CONCLUSIONS: MXF treatment was well tolerated in children with cIAIs. However, a lower clinical cure rate was observed with MXF treatment compared with COMP. This study does not support a recommendation of MXF for children with cIAIs when alternative more efficacious antibiotics with better safety profile are available.

Intussusception: incidence and treatment-insights from the nationwide German surveillance.

OBJECTIVE: Intussusception (IS) is one of the most common paediatric emergencies, and the best mode of conservative reduction and its exact incidence remains unclear. For different reasons, availability of reliable incidence data are useful and additionally may be fundamental to monitor potential effects of recently introduced rotavirus (RV) vaccines. METHODS: We performed a prospective German nationwide surveillance between January 1, 2006 and December 31, 2007, followed by separate collection of all IS cases in a random sample of 31 clinics for an unbiased estimation of underreporting. For case definition, the Brighton Collaboration (BC) criteria were applied. RESULTS: A total of 1200 children with at least 1 episode of IS were included. For children younger than 1 year the incidence was calculated to be 60.4/100,000 child-years. The risk for surgery increased 2-fold if the interval between onset of symptoms and first attempt of conservative reduction exceeded 5 hours (95% confidence interval [CI] 1.2-3.1). We also observed a 2.8-fold increased risk for surgery for hydrostatic (CI 1.2-6.4) and a 3.7-fold for barium enema reduction (CI 1.6-8.8) compared to pneumatic reduction. The level of specialisation of the hospital did not influence the success of conservative management. CONCLUSIONS: For children with IS a fast attempt of pneumatic reduction seems to be the optimal management. Considering the current practice we estimated that approximately 104 (CI 46-161) surgical interventions would be preventable in Germany every year. Also, conduction of reliable postmarketing monitoring of the new RV vaccines is now possible based on the provided incidence data.