Rapid and Robust Continuous Purification of High-Titer Hepatitis B Virus for In Vitro and In Vivo Applications

Available treatments for hepatitis B can control the virus but are rarely curative. This led to a global initiative to design new curative therapies for the 257 million patients affected. Discovery and development of these new therapies is contingent upon functional in vitro and in vivo hepatitis B virus (HBV) infection models. However, low titer and impurity of conventional HBV stocks reduce significance of in vitro infections and moreover limit challenge doses in current in vivo models. Therefore, there is a critical need for a robust, simple and reproducible protocol to generate high-purity and high-titer infectious HBV stocks. Here, we outline a three-step protocol for continuous production of high-quality HBV stocks from supernatants of HBV-replicating cell lines. This purification process takes less than 6 h, yields to high-titer stocks (up to 1 × 1011 enveloped, DNA-containing HBV particles/mL each week), and is with minimal equipment easily adaptable to most laboratory settings.Peer Reviewe

Shallow whole-genome sequencing of plasma cell-free DNA accurately differentiates small from non-small cell lung carcinoma

Background Accurate lung cancer classification is crucial to guide therapeutic decisions. However, histological subtyping by pathologists requires tumor tissue-a necessity that is often intrinsically associated with procedural difficulties. The analysis of circulating tumor DNA present in minimal-invasive blood samples, referred to as liquid biopsies, could therefore emerge as an attractive alternative. Methods Concerning adenocarcinoma, squamous cell carcinoma, and small cell carcinoma, our proof of concept study investigates the potential of liquid biopsy-derived copy number alterations, derived from single-end shallow whole-genome sequencing (coverage 0.1-0.5x), across 51 advanced stage lung cancer patients. Results Genomic abnormality testing reveals anomalies in 86.3% of the liquid biopsies (16/20 for adenocarcinoma, 13/16 for squamous cell, and 15/15 for small cell carcinoma). We demonstrate that copy number profiles from formalin-fixed paraffin-embedded tumor biopsies are well represented by their liquid equivalent. This is especially valid within the small cell carcinoma group, where paired profiles have an average Pearson correlation of 0.86 (95% CI 0.79-0.93). A predictive model trained with public data, derived from 843 tissue biopsies, shows that liquid biopsies exhibit multiple deviations that reflect histological classification. Most notably, distinguishing small from non-small cell lung cancer is characterized by an area under the curve of 0.98 during receiver operating characteristic analysis. Additionally, we investigated how deeper paired-end sequencing, which will eventually become feasible for routine diagnosis, empowers tumor read enrichment by insert size filtering: for all of the 29 resequenced liquid biopsies, the tumor fraction could be increased in silico, thereby "rescuing" three out of five cases with previously undetectable alterations. Conclusions Copy number profiling of cell-free DNA enables histological classification. Since shallow whole-genome sequencing is inexpensive and often fully operational at routine molecular laboratories, this finding has current diagnostic potential, especially for patients with lesions that are difficult to reach

Effects of music therapy in the treatment of children with delayed speech development - results of a pilot study

<p>Abstract</p> <p>Background</p> <p>Language development is one of the most significant processes of early childhood development. Children with delayed speech development are more at risk of acquiring other cognitive, social-emotional, and school-related problems. Music therapy appears to facilitate speech development in children, even within a short period of time. The aim of this pilot study is to explore the effects of music therapy in children with delayed speech development.</p> <p>Methods</p> <p>A total of 18 children aged 3.5 to 6 years with delayed speech development took part in this observational study in which music therapy and no treatment were compared to demonstrate effectiveness. Individual music therapy was provided on an outpatient basis. An ABAB reversal design with alternations between music therapy and no treatment with an interval of approximately eight weeks between the blocks was chosen. Before and after each study period, a speech development test, a non-verbal intelligence test for children, and music therapy assessment scales were used to evaluate the speech development of the children.</p> <p>Results</p> <p>Compared to the baseline, we found a positive development in the study group after receiving music therapy. Both phonological capacity and the children's understanding of speech increased under treatment, as well as their cognitive structures, action patterns, and level of intelligence. Throughout the study period, developmental age converged with their biological age. Ratings according to the Nordoff-Robbins scales showed clinically significant changes in the children, namely in the areas of client-therapist relationship and communication.</p> <p>Conclusions</p> <p>This study suggests that music therapy may have a measurable effect on the speech development of children through the treatment's interactions with fundamental aspects of speech development, including the ability to form and maintain relationships and prosodic abilities. Thus, music therapy may provide a basic and supportive therapy for children with delayed speech development. Further studies should be conducted to investigate the mechanisms of these interactions in greater depth.</p> <p>Trial registration</p> <p>The trial is registered in the German clinical trials register; Trial-No.: DRKS00000343</p

Effects of eight neuropsychiatric copy number variants on human brain structure

Many copy number variants (CNVs) confer risk for the same range of neurodevelopmental symptoms and psychiatric conditions including autism and schizophrenia. Yet, to date neuroimaging studies have typically been carried out one mutation at a time, showing that CNVs have large effects on brain anatomy. Here, we aimed to characterize and quantify the distinct brain morphometry effects and latent dimensions across 8 neuropsychiatric CNVs. We analyzed T1-weighted MRI data from clinically and non-clinically ascertained CNV carriers (deletion/duplication) at the 1q21.1 (n = 39/28), 16p11.2 (n = 87/78), 22q11.2 (n = 75/30), and 15q11.2 (n = 72/76) loci as well as 1296 non-carriers (controls). Case-control contrasts of all examined genomic loci demonstrated effects on brain anatomy, with deletions and duplications showing mirror effects at the global and regional levels. Although CNVs mainly showed distinct brain patterns, principal component analysis (PCA) loaded subsets of CNVs on two latent brain dimensions, which explained 32 and 29% of the variance of the 8 Cohen’s d maps. The cingulate gyrus, insula, supplementary motor cortex, and cerebellum were identified by PCA and multi-view pattern learning as top regions contributing to latent dimension shared across subsets of CNVs. The large proportion of distinct CNV effects on brain morphology may explain the small neuroimaging effect sizes reported in polygenic psychiatric conditions. Nevertheless, latent gene brain morphology dimensions will help subgroup the rapidly expanding landscape of neuropsychiatric variants and dissect the heterogeneity of idiopathic conditions

Frequency and multimorbidity of acute and chronic diseases of different body systems in GP-patients and their impact on participation restrictions

Einleitung: Multimorbidität ist weit verbreitet und stellt ein vorrangiges Problem in Hausarztpraxen dar. Neben der Behandlung von Krankheitssymptomen sind auch Krankheitsfolgen, d. h. Fähigkeitsbeeinträchtigungen und resultierende Teilhabeeinschränkungen zu berücksichtigen. Zur Abschätzung der sozialmedizinischen Versorgungserfordernisse wurde untersucht, welches Erkrankungs- und Komorbiditätsspektrum in Allgemeinarztpraxen vorzufinden und mit welchen sozialen Beeinträchtigungen vorrangig zu rechnen ist. Methode: In einer unausgelesenen Patientenstichprobe aus 40 Hausarztpraxen wurde mittels der Burvill-Skala für zehn Körperregionen erfasst, welche akuten und/oder chronischen Erkrankungen vorliegen. Mit Hilfe der IMET-Skala wurde untersucht, in welchen von zehn Lebensbereichen krankheitsbedingte Einschränkungen vorliegen. Es lagen desweiteren Angaben zu Alter, Geschlecht und Beruf vor. Ergebnisse: Die häufigsten Störungen sind muskuloskelettale (62.5%) und psychische (56.6%) Erkrankungen. 83.0% der Patienten in Allgemeinarztpraxen sind an mindestens zwei, 65.8% an mindestens drei und 29.7% an mindestens fünf verschiedenen Organsystemen (akut und/oder chronisch) erkrankt. Durchschnittlich ist ein Patient an 3.5 (SD 2.0) Organsystemen von Gesundheitsstörungen betroffen. Die Zahl der erkrankten Organsysteme nimmt mit steigendem Alter zu. 30.3% der bis einschließlich 30-Jährigen, 15.1% der 31-60-Jährigen und 8.6% der > 60 Jährigen haben maximal ein erkranktes Organsystem. Frauen sind multimorbider als Männer. 14.5% der Frauen weisen in maximal einem Organsystem Störung(en) auf, 10.3% sind von sieben oder mehr Gesundheitsstörungen betroffen. 20.9% der Männer berichten über maximal ein erkranktes Organsystem, in sieben oder mehr Organsystemen haben 6.5% der Männer Erkrankungen. Am stärksten beeinträchtigt sind die Patienten im Bereich ‚Arbeit/Beruf‘ (MW 3.20, SD 3.28) und ‚außergewöhnlichen Belastungssituationen‘ (MW 2.81, SD 3.05). 40.9% geben im Berufsleben mindestens mittelschwere, 17.6% schwere bis schwerste Beeinträchtigungen an. 38.0% der Patienten beschreiben mindestens mittelschwere Beeinträchtigungen bei der Bewältigung außergewöhnlicher Belastungen. Am wenigsten beeinträchtigt sind sie in ‚üblichen Alltagsaktivitäten‘ (MW 1.35, SD 2.31). Bezogen auf die Individualmorbidität sind Patienten mit Erkrankungen der Sinnesorgane mit einem IMET-Mittelwert von 2.5 (SD 2.5) am wenigsten teilhabebeeinträchtigt, am stärksten neurologisch Erkrankte (IMET-MW 3.8, SD 2.5). Wendet man das Prinzip der DALY an, dann sind psychische Erkrankungen (1.69) in Hausarztpraxen sozialmedizinisch von höchster Relevanz, an zweiter Stelle folgen muskuloskelettale Erkrankungen (1.62). Neurologische Erkrankungen haben auf Grund ihrer niedrigen Prävalenz die geringste Bedeutung (0.31). Schlussfolgerung: Multimorbidität ist die Regel in der Allgemeinarztpraxis. Psychische Erkrankungen haben vor muskuloskelettalen Erkrankungen die größte sozialmedizinische Relevanz. Insbesondere psychische und neurologische Erkrankungen führen zu relevanten Teilhabestörungen.Background: Multimorbidity is a common problem, especially in general practice. It is important to treat not only symptoms of diseases, but also resulting disorders of function and participation. Aim of this study was to investigate the spectrum of disorders, the types of comorbidity, and the rates of impairment in GP-patients and to evaluate their medical and social needs of care. Method: A convenience sample of patients from 40 general practitioners filled in the Burvill-Scale and the IMET-Scale. The Burvill-Scale asks for acute and chronic illnesses in ten different body regions. The IMET-Scale asks for illness-based participation restrictions in ten areas of life. Additional information was available on patient age, gender, education and occupational status. Results: Most frequent were musculoskeletal (62.5%) and psychological problems (56.6%). There were 83.0% of GP-patients with at least two, 65.8% with at least three and 29.7% with at least five acute and/or chronic disorders in different body systems. On average patients reported 3.5 (SD 2.0) affected body systems. Older patients and women report more morbidity. 30.3 % of patients aged 30 or younger report no or only one health problem. 15.1% of patients in the age of 31 to 60 and 8.5% aged 61 or older say that there is no or only one problem. 14.5% of female patients report no or only one problem, and 10.3% seven or more. 20.9% males report no or only one problem and 6.5% seven or more. Patients feel most impaired in the areas ‘work’ (IMET-MW 3.20, SD 3.28) and ‘coping with special demands’ (MW 2.81, SD 3.05). 40.9% are at least moderately severe, 17.6% severe or very severely impaired at work. 38.0% are at least moderately severe impaired in ‘coping with special demands’. In the domain ‘general activities in daily life’ patients are least affected (MW 1.35, SD 2.31). In respect to selected illnesses ‘eye/ear’ show least participation restrictions (IMET-MW 2.5, SD 2.5), patients with neurological diseases are most affected (IMET-MW 3.8, SD 2.5). Referring to the DALY concept, psychological illnesses (1.69) are most important for the general practitioner, musculoskeletal diseases come second (1.62). Because of their low prevalence neurological diseases are of little importance (0.31). Conclusion: Multimorbidity in GP-patients is the rule, not the exception. Psychological and musculoskeletal problems have the greatest impact on life. Especially psychological and neurological health problems lead to relevant participation restrictions

Assessment of pressure-volume relations in univentricular hearts: Comparison of obtainment by real-time 3D echocardiography and mini pressure-wire with conductance technology.

ObjectivesThe gold standard to obtain pressure-volume relations (PVR) of the heart, the conductance technology (PVRCond), is rarely used in children. PVR can also be obtained by 3D-echocardiography volume data combined with simultaneously measured pressure data by a mini pressure-wire (PVR3DE). We sought to investigate the feasibility of both methods in patients with univentricular hearts and to compare them, including hemodynamic changes.MethodsWe studied 19 patients (age 2-29 years). PVR3DE and PVRCond were assessed under baseline conditions and stimulation with dobutamine.ResultsObtaining PVR3DE was successful in all patients. Obtaining PVRCond was possible in 15 patients during baseline (79%) and in 12 patients under dobutamine (63%). Both methods showed that end-systolic elastance (Ees) and arterial elastance (Ea) increased under dobutamine and that Tau showed a statistically significant decrease. Intraclass correlation (95% confidence interval) showed moderate to good agreement between methods: Ees: 0.873 (0.711-0.945), Ea: 0.709 (0.336-0.873), Tau: 0.867 (0.697-0.942). Bland-Altman analyses showed an acceptable bias with wider limits of agreement: Ees: 1.63 mmHg/ml (-3.83-7.08 mmHg/ml), Ea: 0.53 mmHg/ml (-5.23-6.28 mmHg/ml), Tau: -0,76 ms (-10.73-9.21 ms).ConclusionChanges of PVR-specific parameters under dobutamine stimulation were reflected in the same way by both methods. However, the absolute values for these parameters could vary between methods and, therefore, methods are not interchangeable. Obtaining PVR3DE in a single ventricle was easier, faster and more successful than PVRCond. PVR3DE provides a promising and needed alternative to the conductance technology for the assessment of cardiac function in univentricular hearts

The Embittered Mind. Dimesions of Embitterment and Validation of the Concept

There is a growing interest in embitterment as psychological concept. However, little systematic research has been conducted to characterize this emotional reaction. Still, there is an ongoing debate about the distinctiveness of embitterment and its dimensions. Additionally, a categorical and a dimensional perspective on embitterment have been developed independently over the last decade. The present study investigates the dimensions of embitterment by bringing these two different approaches together, for the first time. The Bern Embitterment Inventory (BEI) was given to 49 patients diagnosed with “Posttraumatic Embitterment Disorder (PTED)” and a matched control group of 49 patients with psychological disorders with other dominant emotional dysregulations. The ability to discriminate between the two groups was assessed by t-tests and Receiver Operating Characteristic Curves (ROC curve analysis). PTED patients scored significantly higher on the BEI than the patients of the control group. ROC analyses indicated diagnostic accuracy of the inventory. Further, we conducted Confirmatory Factor Analyses (CFA) to examine the different dimensions of embitterment and their relations. As a result, we found four characteristic dimensions of embitterment, namely disappointment, lack of acknowledge, pessimism, and misanthropy. In general, our findings showed a common understanding of embitterment as a unique but multidimensional emotional reaction to distressful life-events