Development of a CENH3-based and non-viral vector system

New vectors for successful transgene delivery in patients are more than needed. Current gene therapeutic vectors are mostly based on virus genomes. A main reason is the very efficient administration of the vector into the target cells. The viral background of these systems also has severe side effects like e.g. immunological reactions of the patient or the activation of oncogenes due to insertional mutagenesis, which can result in cancer development. A possibility to overcome these problems is the establishment of extrachromosomally maintained, autonomously replicating and non-viral vectors. The goal of this work was to establish such a novel system with the help of the CENH3 protein. The centromere-specific histone 3 variant CENH3 (H.sapiens: CENH3CENP-A, D.melanogaster: CENH3CID) is sufficient for centromere formation. It confers a stable and epigenetically heritable mark at the centromeric region and does this also on plasmids. With the help of CENH3 I changed the passive piggyback segregation mechanism of Epstein-Barr virus-derived vectors into an active segregation mechanism for plasmid DNA (pDNA) vectors. I demonstrated that the new and active pDNA vector segregation mechanism prolongs pDNA vector retention in cells. The information gained from basic research might have great impact in the field of gene-therapeutic research, as this mechanism might be a helpful tool in future gene therapeutic vectors.Neue Weiterentwicklungen von Genvektoren für die Gentherapie sind dringend notwendig. Die meisten, derzeit in der Klinik angewandten, Genvektoren basieren auf Virusgenomen. Durch Infektion mit virusähnlichen Partikeln wird ein besonders effizienter Gentransport in die Zielzelle erreicht. Der virale Hintergrund bringt jedoch negative Effekte wie z.B. immunologische Reaktionen des Patienten oder die Aktivierung von Onkogenen durch Insertionsmutagenese und somit Krebsentstehung, mit sich. Eine Möglichkeit, die Probleme zu umgehen sind extrachromosomale, autonom replizierende und nicht-virale Genvektoren. Ziel dieser Arbeit war die Entwicklung eines neuen Genvektorsystems, dessen extrachromosomale Stabilität auf dem Protein CENH3 basiert. Das Zentromer-spezifische Histon 3 CENH3 (H.sapiens: CENH3CENP-A, D.melanogaster: CENH3CID) ist alleine ausreichend für die Zentromerausbildung. CENH3 etabliert eine stabile, epigenetisch vererbte Markierung auf dem Chromosom sowie auf Plasmiden. Mit Hilfe von CENH3 war es möglich den passiven Huckepack-Mechanismus von Epstein-Barr-Virus basierten Plasmiden in einen aktiven Segregationsprozess für Plasmid-DNA Vektoren zu verändern. Durch den neuen, aktiven Segregationsmechanismus wird die Vektorstabilität in der Zielzelle erhöht. Die Grundlagenforschung liefert uns hier wertvolle Informationen, die äußerst hilfreich für die Entwicklung neuer Genvektoren ist

How important is the Family? : Alpha nuclear potentials and p-process nucleosynthesis

Copyright owned by the author(s) under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike LicenceIn this work we present the results from the analysis of the experimentally measured angular distributions of the reaction 106Cd(α , α )106 Cd at several different energies around the Coulomb barrier. The difficulties that arise in the study of 106Cd-α -nuclear potential and the so called Family Problem are addressed in this work

Configural frequency analysis as a method of determining patients' preferred decision-making roles in dialysis

<p>Abstract</p> <p>Background</p> <p>Numerous studies examined factors in promoting a patient preference for active participation in treatment decision making with only modest success. The purpose of this study was to identify types of patients wishing to participate in treatment decisions as well as those wishing to play a completely active or passive role based on a Germany-wide survey of dialysis patients; using a prediction typal analysis method that defines types as configurations of categories belonging to different attributes and takes particularly higher order interactions between variables into account.</p> <p>Methods</p> <p>After randomly splitting the original patient sample into two halves, an exploratory prediction configural frequency analysis (CFA) was performed on one-half of the sample (n = 1969) and the identified types were considered as hypotheses for an inferential prediction CFA for the second half (n = 1914). 144 possible prediction types were tested by using five predictor variables and control preferences as criterion. An α-adjustment (0.05) for multiple testing was performed by the Holm procedure.</p> <p>Results</p> <p>21 possible prediction types were identified as hypotheses in the exploratory prediction CFA; four patient types were confirmed in the confirmatory prediction CFA: patients preferring a passive role show low information seeking preference, above average trust in their physician, perceive their physician's participatory decision-making (PDM)-style positive, have a lower educational level, and are 56-75 years old (Type 1; <it>p </it>< 0.001) or > 76 years old (Type 2; <it>p </it>< 0.001). Patients preferring an active role show high information seeking preference, a higher educational level, and are < 55 years old. They have either below average trust, perceive the PDM-style negative (Type 3; <it>p </it>< 0.001) or above average trust and perceive the PDM-style positive (Type 4; <it>p </it>< 0.001).</p> <p>Conclusions</p> <p>The method prediction configural frequency analysis was newly introduced to the research field of patient participation and could demonstrate how a particular control preference role is determined by an association of five variables.</p

Importance of hemodialysis-related outcomes: comparison of ratings by a self-help group, clinicians, and health technology assessment authors with those by a large reference group of patients

Janssen IM, Scheibler F, Gerhardus A. Importance of hemodialysis-related outcomes: comparison of ratings by a self-help group, clinicians, and health technology assessment authors with those by a large reference group of patients. Patient Preference and Adherence. 2016;10:2491-2500.Background: The selection of important outcomes is a crucial decision for clinical research and health technology assessment (HTA), and there is ongoing debate about which stakeholders should be involved. Hemodialysis is a complex treatment for chronic kidney disease (CKD) and affects many outcomes. Apart from obvious outcomes, such as mortality, morbidity and health-related quality of life (HRQoL), others such as, concerning daily living or health care provision, may also be important. The aim of our study was to analyze to what extent the preferences for patient-relevant outcomes differed between various stakeholders. We compared preferences of stakeholders normally or occasionally involved in outcome prioritization (patients from a self-help group, clinicians and HTA authors) with those of a large reference group of patients. Participants and methods: The reference group consisted of 4,518 CKD patients investigated previously. We additionally recruited CKD patients via a regional self-help group, nephrologists via an online search and HTA authors via an expert database or personal contacts. All groups assessed the relative importance of the 23 outcomes by means of a discrete visual analog scale. We used descriptive statistics to rank outcomes and compare the results between groups. Results: We received completed questionnaires from 49 self-help group patients, 19 nephrologists and 18 HTA authors. Only the following 3 outcomes were ranked within the top 7 outcomes by all 4 groups: safety, HRQoL and emotional state. The ratings by the self-help group were generally more concordant with the reference group ratings than those by nephrologists, while HTA authors showed the least concordance. Conclusion: Preferences of CKD patients from a self-help group, nephrologists and HTA authors differ to a varying extent from those of a large reference group of patients with CKD. The preferences of all stakeholders should form the basis of a transparent approach so as to generate a valid list of important outcomes

A scattered landscape: assessment of the evidence base for 71 patient decision aids developed in a hospital setting

Background Recent publications reveal shortcomings in evidence review and summarization methods for patient decision aids. In the large-scale "Share to Care (S2C)" Shared Decision Making (SDM) project at the University Hospital Kiel, Germany, one of 4 SDM interventions was to develop up to 80 decision aids for patients. Best available evidence on the treatments' impact on patient-relevant outcomes was systematically appraised to feed this information into the decision aids. Aims of this paper were to (1) describe how PtDAs are developed and how S2C evidence reviews for each PtDA are conducted, (2) appraise the quality of the best available evidence identified and (3) identify challenges associated with identified evidence. Methods The quality of the identified evidence was assessed based on GRADE quality criteria and categorized into high-, moderate-, low-, very low-quality evidence. Evidence appraisal was conducted across all outcomes assessed in an evidence review and for specific groups of outcomes, namely mortality, morbidity, quality of life, and treatment harms. Challenges in evidence interpretation and summarization resulting from the characteristics of decision aids and the type and quality of evidence are identified and discussed. Conclusions Evidence reviews in this project were carefully conducted and summarized. However, the evidence identified for our decision aids was indeed a "scattered landscape" and often poor quality. Facing a high prevalence of low-quality, non-directly comparative evidence for treatment alternatives doesn't mean it is not necessary to choose an evidence-based approach to inform patients. While there is an urgent need for high quality comparative trials, best available evidence nevertheless has to be appraised and transparently communicated to patients

Making shared decision-making (SDM) a reality: protocol of a large-scale long-term SDM implementation programme at a Northern German University Hospital

Introduction: Shared decision-making (SDM) is not yet widely used when making decisions in German hospitals. Making SDM a reality is a complex task. It involves training healthcare professionals in SDM communication and enabling patients to actively participate in communication, in addition to providing sound, easy to understand information on treatment alternatives in the form of evidence-based patient decision aids (EbPDAs). This project funded by the German Innovation Fund aims at designing, implementing and evaluating a multicomponent, large-scale and integrative SDM programme-called SHARE TO CARE (S2C)-at all clinical departments of a University Hospital Campus in Northern Germany within a 4-year time period. Methods and analysis S2C tackles the aforementioned components of SDM: (1) training physicians in SDM communication, (2) activating and empowering patients, (3) developing EbPDAs in the most common/relevant diseases and (4) training other healthcare professionals in SDM coaching. S2C is designed together with patients and providers. The physicians' training programme entails an online and an in situ training module. The decision coach training is based on a similar but less comprehensive approach. The development of online EbPDAs follows the International Patient Decision Aid Standards and includes written, graphical and video-based information. Validated outcomes of SDM implementation are measured in a preintervention and postintervention evaluation design. Process evaluation accompanies programme implementation. Health economic impact of the intervention is investigated using a propensity-score-matched approach based on potentially preference-sensitive hospital decisions. Ethics and dissemination Ethics committee review approval has been obtained from Medical Ethics Committee of the Medical Faculty of the Christian-Albrechts-University Kiel. Project information and results will be disseminated at conferences, on project-hosted websites at University Hospital Medical Center Schleswig Holstein and by S2C as well as in peer-reviewed and professional journals

Preferences of patients undergoing hemodialysis - results from a questionnaire-based study with 4,518 patients

Janssen IM, Gerhardus A, von Gersdorff G, et al. Preferences of patients undergoing hemodialysis - results from a questionnaire-based study with 4,518 patients. Patient Preference and Adherence. 2015;2015(9):847-855.Background: Chronic kidney disease is an increasing health problem worldwide and in its final stage (stage V) can only be treated by renal replacement therapy, mostly hemodialysis. Hemodialysis has a major influence on the everyday life of patients and many patients report dissatisfaction with treatment. Little is known about which aspects of treatment are considered important by hemodialysis patients. The objective of this study was to rate the relative importance of different outcomes for hemodialysis patients and to analyze whether the relative importance differed among subgroups of patients. Patients and methods: Within the framework of a yearly questionnaire which is distributed among patients receiving hemodialysis by the largest hemodialysis provider in Germany, we assessed the relative importance of 23 outcomes as rated on a discrete visual analog scale. Descriptive statistics were used to rank the outcomes. Subgroup analyses were performed using Mann–Whitney U or Kruskal–Wallis tests. Results: Questionnaires of 4,518 hemodialysis patients were included in the analysis. The three most important outcomes were safety of treatment, health-related quality of life, and satisfaction with care. Further important outcomes were hospital stays, accompanying symptoms, hemodialysis duration, and the improvement or preservation of a good emotional state. Age, profession, and education had the strongest influence on relevant differences of preferences for outcomes; no relevant influence of sex or comorbidity was observed. Conclusion: Outcomes concerning the delivery or provision of care and aspects influencing quality of life are rated by patients to be at least as important as clinical outcomes. Many of the outcomes judged to be important by the patients are not regularly considered in research, evaluation studies, or quality programs

Ethical issues in autologous stem cell transplantation (ASCT) in advanced breast cancer: A systematic literature review

BACKGROUND: An effectiveness assessment on ASCT in locally advanced and metastatic breast cancer identified serious ethical issues associated with this intervention. Our objective was to systematically review these aspects by means of a literature analysis. METHODS: We chose the reflexive Socratic approach as the review method using Hofmann's question list, conducted a comprehensive literature search in biomedical, psychological and ethics bibliographic databases and screened the resulting hits in a 2-step selection process. Relevant arguments were assembled from the included articles, and were assessed and assigned to the question list. Hofmann's questions were addressed by synthesizing these arguments. RESULTS: Of the identified 879 documents 102 included arguments related to one or more questions from Hofmann's question list. The most important ethical issues were the implementation of ASCT in clinical practice on the basis of phase-II trials in the 1990s and the publication of falsified data in the first randomized controlled trials (Bezwoda fraud), which caused significant negative effects on recruiting patients for further clinical trials and the doctor-patient relationship. Recent meta-analyses report a marginal effect in prolonging disease-free survival, accompanied by severe harms, including death. ASCT in breast cancer remains a stigmatized technology. Reported health-related-quality-of-life data are often at high risk of bias in favor of the survivors. Furthermore little attention has been paid to those patients who were dying. CONCLUSIONS: The questions were addressed in different degrees of completeness. All arguments were assignable to the questions. The central ethical dimensions of ASCT could be discussed by reviewing the published literature

Erratum to: Methods for evaluating medical tests and biomarkers

[This corrects the article DOI: 10.1186/s41512-016-0001-y.]

Erratum to: Methods for evaluating medical tests and biomarkers

[This corrects the article DOI: 10.1186/s41512-016-0001-y.]