Insights into the molecular basis of beet curly top resistance in sugar beet through a transcriptomic approach at the early stage of symptom development.

Curly top disease caused by Beet curly top virus (BCTV) is a limiting factor for sugar beet production. The most economical and sustainable control of BCTV in sugar beet would be via the growth of resistant cultivars, although most commercial cultivars possess only low-to-moderate quantitative resistance. A double haploid line (KDH13) showed a high level of resistance to BCTV infection. However, the mechanism of resistance and response of this line to BCTV infection is unknown. Here, we tested the response of this line to both local and systemic BCTV infections. The virus replicated at a high level in locally infected tissue but lower than in susceptible KDH19 plants. Resistant KDH13 plants systemically infected with BCTV showed only mild enation without leaf curling after 30 days. In contrast, severe leaf curling appeared after 12 days in susceptible plants with higher virus accumulation. Transcriptome analysis of the BCTV-infected KDH13 plants at the early stage of symptom development showed only 132 genes that were exclusively deregulated compared to the regulation of a large number of genes (1018 genes) in KDH19 plants. Pathway enrichment analysis showed that differentially expressed genes were predominantly involved in hormone metabolism, DNA methylation, immune response, cell cycle, biotic stress and oxidative stress. The auxin level in both resistant and susceptible plants increased in response to BCTV infection. Remarkably, exogenous application of auxin caused leaf curling phenotype in the absence of the virus. This study demonstrates the response of resistant and susceptible plants to BCTV infection at both local and systemic infections and highlights the defence-related genes and metabolic pathways including auxin for their contribution towards BCTV symptom development and resistance in sugar beet.

Identifying missing links in the conceptualization of financial toxicity: a qualitative study.

PURPOSE: Financial toxicity can have a major impact on the quality of life of cancer survivors but lacks conceptual clarity and understanding of the interrelationships of the various aspects that constitute financial toxicity. This study aims to extract major drivers and mediators along the pathway from cancer-related costs to subjective financial distress from the patients' experiences to establish a better understanding of financial toxicity as a patient-reported outcome. METHODS: Qualitative semistructured interviews with 39 cancer patients were conducted in Germany and addressed patient experiences with cancer-related financial burden and distress in a country with a statutory health care system. Transcripts were analyzed using content analysis. RESULTS: Several aspects of financial burden need to be considered to understand financial toxicity. The assessment of the ability to make ends meet now or in the future and the subjective evaluation of financial adjustments-namely, the burden of applied financial adjustments and the availability of financial adjustment options-mediate the connection between higher costs and subjective financial distress. Moreover, bureaucracy can influence financial distress through a feeling of helplessness during interactions with authorities because of high effort, non-traceable decisions, or one's own lack of knowledge. CONCLUSION: We identified four factors that mediate the impact of higher costs on financial distress that should be addressed in further studies and targeted by changes in policies and support measures. Financial toxicity is more complex than previously thought and should be conceptualized and understood more comprehensively in measurements, including the subjective assessment of available adjustment options and perceived burden of financial adjustments.

Coping mechanisms for financial toxicity: a qualitative study of cancer patients' experiences in Germany.

PURPOSE: The coping mechanisms employed to deal with objective financial burden following a cancer diagnosis are considered mediators of financial toxicity, specifically for the relationship between increased costs and health outcomes. Using qualitative research, the aim of the study was to explore the financial adjustments experienced by cancer patients and identify key coping mechanisms. METHODS: Semistructured interviews with 39 cancer patients (aged from 40 to 86 years) were conducted between May 2017 and April 2018 in Germany. Narratives were transcribed verbatim and analysed via qualitative content analysis. RESULTS: In all, 28 participants reported that they made financial adjustments related to two major categories: (1) reducing expenditures related to basic needs, luxury needs, and health-related decisions, and (2) increasing financial resources by saving less, using savings, investing manpower, obtaining help from third parties, incurring bank debt, and making health-related decisions. Typically, cancer patients both reduced expenditures and increased financial resources, but no typical patterns of combinations of coping mechanisms could be identified. However, reducing spending on basic and luxury needs was found to occur more typically than increasing the available money through incurring bank debt, making health-related decisions, or investing manpower. CONCLUSIONS: This qualitative study provides a comprehensive understanding of the complexity of the coping mechanisms used by patients to address either higher costs or changed needs and priorities following a cancer diagnosis. With regard to understanding patients' experiences of subjective financial stress, both increasing financial resources and reducing expenditures, particularly those relating to basic needs and luxury needs as distinct categories, might be relevant and should be considered. The financial situation impacts the treatment of cancer patients since health-related decisions were reported to be made with the aim of increasing financial resources by maintaining an ability to work or reducing expenditures through non-adherence.

["If It wouldn't be Made So Difficult for Us" - Barriers on Access to Pediatric Rehabilitation and Potential for Optimization from the Pediatricians' Perspective]. / "Wenn es uns nicht so schwer gemacht werden würde" ­ Zugangsbarrieren und Optimierungspotenziale zur Kinder- und Jugendrehabilitation aus Perspektive der Kinder- und Jugendärzte.

OBJECTIVE: The study explores barriers impeding the access to pediatric rehabilitation and potential for optimization from the pediatricians' perspective. METHODS: Semi-structured expert interviews (N=25) and focus groups (N=3) with pediatricians were conducted and analyzed according to the method of Meuser & Nagel. The results were discussed within an expert group consisting of representatives of the statutory pension insurance and the professional association of pediatricians. RESULTS: Barriers appear due to information deficits regarding carrier-oriented access criteria. A perceived time-consuming application, high refusal rates with insufficient reasons and a lack of feedback from the carrier to the physician inhibit the application motivation. Mentioned improvements refer to a direct feedback of the application decision to the physician, case-specific reasons for refusal, physician oriented information as well as an increased presence of rehabilitation at pediatric congresses and medical trainings. CONCLUSION: To strengthen the role of rehabilitation in pediatric care improved information and knowledge in the allocation and submission of rehabilitation applications of primary care physicians is needed as well as an enhanced communication between rehabilitation carriers and physicians.

Improving Rehabilitative Care of Methamphetamine Users in Germany: The Expert's Perspective.

Methamphetamine is one of the most frequently used drugs worldwide. In Germany, methamphetamine use has greatly increased in recent years, presenting the rehabilitative treatment system with new challenges. In this study, we identified deficits and possibilities for optimization in the field of medical rehabilitation. A total of 39 interviews and two focus groups with experts along the treatment course of methamphetamine users were conducted. Our analyses indicate that methamphetamine users are more difficult to treat compared with patients who consume other drugs. They are more likely to be associated with problematic characteristics and behaviors than other rehabilitants. Several health care deficits were revealed: too short rehabilitation treatment, no specific or differentiated therapy concepts, lack of capacity for education and vocational training, lack of outpatient options, and insufficient facilities for parents and children. Findings indicate that inadequate rehabilitation is being provided for methamphetamine users in Germany, indicating a need to adapt treatment for this group.

[Methamphetamine Use in Central Germany: A Qualitative Study on Consumer Groups and Motives from the Experts' Perspective]. / Methamphetaminkonsum in Mitteldeutschland: Eine qualitative Studie zu Konsumentengruppen und -motiven aus Expertensicht.

BACKGROUND: Methamphetamine is one of the most consumed illegal drugs worldwide. In Germany, Methamphetamine shows the highest rates of growth in comparison to other illegal substances in recent years. Particularly Central Germany has been struck by a high rise in consumption. International studies indicate that there is no specific group of people that can be identified as Methamphetamine users. Different consumption patterns in terms of Methamphetamine use can be identified within various social groups. This qualitative study will explore different consumer groups among methamphetamine users, their motives, and how they differ by drug addiction type. METHODS: The empirical data collection was carried out in a consecutive 2-stage process. Initially, 39 semi-structured individual interviews with experts from different clinical areas were carried out about their experience and perspectives on Methamphetamine use. The results of the individual interviews were subsequently discussed and validated within 2 interdisciplinary focus groups. All interviews and focus groups were digitally recorded, transcribed and analyzed according to the method of Meuser & Nagel. RESULTS: Altogether, 3 consumer groups were identified: (1) Young parents, women and pregnant women, (2) young drug users and early adapters and (3) older drug users or late entrants. The guiding motive in terms of Methamphetamine use described by the experts was improved efficiency. Further motives are for instance overcoming stressful situations, enablement and improvement of sexual experiences, self-esteem enhancement, coping with crisis or trauma, curiosity and drug use as leisure time activity. DISCUSSION/CONCLUSION: The results in terms of consumer groups and consumption motives are consistent with international findings and we were able to verify and expand them for Central Germany. The outcomes illustrated that there are different consumer groups among methamphetamine users that differ from consumer groups of other drug addiction types. Treatment and consultation have to anticipate these challenges and adapt their strategies to different needs. Finally, further improvement of accessibility for those affected persons is an imperative to the German healthcare system.

[Access to Inpatient Rehabilitation of Methamphetamine Addiction - Barriers and Potential for Improvement from the Experts' Perspective]. / Zugang zur stationären Rehabilitation bei Methamphetaminabhängigkeit ­ Barrieren und Optimierungspotenziale aus Expertenperspektive.

OBJECTIVE: The study explores central barriers impeding the access to inpatient rehabilitation of methamphetamine addicts and investigates how these barriers can be tackled. METHODS: The experiences and perspectives of experts from different healthcare sectors were explored within 39 semi-structured individual interviews. Subsequently, the results gathered in the course of the individual interviews were discussed within 2 inter-professional focus groups. All interviews and focus group discussions were analyzed according to the interpretive analysis method of Meuser & Nagel. RESULTS: Structural issues in particular impair the access to inpatient rehabilitation of methamphetamine dependents. This includes impeded access to rehabilitation via general practitioners, rare implementation of seamless transitions, capacity problems as well as non-transparent procedures in terms of treatment facility allocations. CONCLUSION: Various barriers impair access to inpatient rehabilitation of methamphetamine dependents. It is important to increase awareness of methamphetamine addicts and their specific needs particularly among general practitioners. Furthermore, it is important to ensure seamless transitions from detoxification treatment to inpatient rehabilitation and to reduce waiting periods as well as to provide well-differentiated and appropriate rehabilitation options.

Socioeconomic differences in the pathways to diagnosis of coronary heart disease: a qualitative study.

Background: Socioeconomic inequalities in coronary heart disease (CHD)-related morbidity and mortality are well explored. However, less is known about the causes of inequalities in CHD treatment. In this qualitative study, we explored socioeconomic differences in the pathways to diagnosis of CHD. Methods: The data originated from 38 semi-structured interviews with older CHD patients, aged 59-80 years, conducted at the university hospital in Halle, Germany, between November 2014 and April 2015. We analysed the narratives related to the time before CHD was confirmed by coronary angiography electively or urgently. Transcripts were analysed following inductive qualitative content analysis and we identified socioeconomic differences by comparing and contrasting patients' narratives. Results: The patients interpreted their symptoms based on expectations, normalization, relief and obtaining help from third parties. For those experiencing chronic CHD symptoms, only patients with low socioeconomic status (SES) waited to seek healthcare until they suffered myocardial infarction. Mainly low-SES patients procrastinated in undergoing diagnostic procedures. We found no socioeconomic differences in the urgent pathway. However, along the elective pathway, only low-SES patients reported receiving assistance from a general practitioner in accessing a cardiologist. Conclusions: Socioeconomic differences in CHD diagnosis were mainly apparent before patients sought healthcare. These differences were more pronounced when CHD was electively diagnosed due to chronic symptoms rather than urgently diagnosed due to acute symptoms. To address socioeconomic differences, general practitioners should focus on any indication of symptoms and interpretation mentioned by low-SES patients, and coordinate these patients' pathways to diagnosis while emphasizing the seriousness of CHD.

Molecular evolution and selection patterns of plant F-box proteins with C-terminal kelch repeats.

The F-box protein superfamily represents one of the largest families in the plant kingdom. F-box proteins phylogenetically organize into numerous subfamilies characterized by their carboxyl (C)-terminal protein-protein interaction domain. Among the largest F-box protein subfamilies in plant genomes are those with C-terminal kelch repeats. In this study, we analyzed the phylogeny and evolution of F-box kelch proteins/genes (FBKs) in seven completely sequenced land plant genomes including a bryophyte, a lycophyte, monocots, and eudicots. While absent in prokaryotes, F-box kelch proteins are widespread in eukaryotes. Nonplant eukaryotes usually contain only a single FBK gene. In land plant genomes, however, FBKs expanded dramatically. Arabidopsis thaliana, for example, contains at least 103 F-box genes with well-conserved C-terminal kelch repeats. The construction of a phylogenetic tree based on the full-length amino acid sequences of the FBKs that we identified in the seven species enabled us to classify FBK genes into unstable/stable/superstable categories. In contrast to superstable genes, which are conserved across all seven species, kelch domains of unstable genes, which are defined as lineage specific, showed strong signatures of positive selection, indicating adaptational potential. We found evidence for conserved protein features such as binding affinities toward A. thaliana SKP1-like adaptor proteins and subcellular localization among closely related FBKs. Pseudogenization seems to occur only rarely, but differential transcriptional regulation of close relatives may result in subfunctionalization.

Targeted mutagenesis in plants using Beet curly top virus for efficient delivery of CRISPR/Cas12a components.

Genome editing using CRISPR/Cas is rapidly being developed for gene targeting in eukaryotes including plants. However, gene targeting by homology-directed DNA recombination (HDR) is an infrequent event compared to the dominant DNA repair by non-homologous end-joining. Another bottleneck is the ineffective delivery of CRISPR/Cas components into plant cells. To overcome these constraints, here a geminiviral replicon from Beet curly top virus (BCTV) has been produced with a wide host range and high DNA accumulation capacity for efficient delivery of CRISPR/Cas12a components into plant cells. Initially, a BCTV replicon was prepared after removing the virion sense genes from an infectious full-length clone for agrobacterium mediated infection. This replicon expressed a green fluorescent protein (GFP) marker gene at a high level compared to T-DNA binary vector. In transient assay, the BCTV replicon produced a higher rate of mutagenesis and HDR in the GFP transgene in Nicotiana benthamiana through efficient delivery of CRISPR/Cas12a components compared to the cognate T-DNA control. This was through a range of complete or partial HDR for conversion of GFP into YFP after exchange of a single amino acid (Thr224Tyr) in the target gene. In addition, induced mutagenesis and HDR in the target gene were heritable. Thus, the BCTV replicon provides a new tool for efficient delivery of CRISPR/Cas12a components that could be used in a wide range of dicotyledonous plants. The established GFP to YFP system and the GFP mutant line produced also enable further optimization and understanding of HDR in plants via CRISPR/Cas12a system using geminiviral replicons.

'It was a big monetary cut'-A qualitative study on financial toxicity analysing patients' experiences with cancer costs in Germany.

Receiving information about expected costs promptly after a cancer diagnosis through psycho-oncology care or social counselling is crucial for patients to be prepared for the financial impact. Nevertheless, less is known about financial impacts for cancer patients in countries with statutory health insurance. This study aims to explore the full scope of costs that constitute the financial impact of a cancer diagnosis in Germany and to identify the reasons for high financial decline. Semistructured interviews with 39 cancer patients were conducted between May 2017 and April 2018. Narratives were analysed via qualitative content analysis. Several factors influenced cancer patients' indirect costs and direct medical and non-medical costs. For many patients, these changes resulted in higher indirect costs caused by income losses, especially when surcharges for shift work, travel expenses or company benefits ceased and were not reimbursed. Higher direct medical costs were caused by co-payments and additional non-refundable costs. Non-medical costs were reported to increase for some patients and to decrease for others, as for example, leisure activity costs either increasing because of pampering oneself to cope with the diagnosis and undergoing therapy or decreasing because of not being able to participate in leisure activities during therapy. When analysing the financial impacts of individuals' total costs, we found that some patients experienced no financial decline or an overall financial increase. Most patients experienced overall higher costs, and income loss was the main driver of a high financial decline. Nevertheless, decreased non-medical costs due to lower work-related and leisure activity costs could compensate for these higher costs. Cancer patients are confronted with a variety of changes in their financial situations, even in countries with statutory health insurance. Screening for cancer patients with a high risk of financial decline should consider any effects on indirect costs and direct medical and nonmedical costs.

Methamphetamine use in Central Germany: protocol for a qualitative study exploring requirements and challenges in healthcare from the professionals' perspective.

INTRODUCTION: The synthetic drug methamphetamine with its high addiction potential is associated with substantial adverse health effects. In Germany, especially Central Germany, the increase in the consumption of methamphetamine has exceeded that of other illegal drugs. The treatment system and service providers are facing new challenges due to this rise in consumption. This qualitative study will explore the demand created by the increasing healthcare needs of methamphetamine-addicted persons in Central Germany, and the difficulty of rehabilitating addicted people. METHODS AND ANALYSIS: The collection of empirical data will take place in a consecutive, two-stage process. In the first part of data collection, the experiences and perspectives of 40 professionals from numerous healthcare sectors for methamphetamine-addicted persons will be explored with the help of semistructured face-to-face interviews and probed by the research team. These findings will be discussed in 2 focus groups consisting of the participants of the face-to-face interviews; these group discussions comprise the second part of the data collection process. The interviews will be audio recorded, transcribed, and then subjected to qualitative content analysis. ETHICS AND DISSEMINATION: All interviewees will receive comprehensive written information about the study, and sign a declaration of consent prior to the interview. The study will comply rigorously with data protection legislation. The research team has obtained the approval of the Ethical Review Committee at the Martin Luther University Halle-Wittenberg, Germany. The results of the study will be published in high-quality, peer-reviewed international journals, presented at several congresses and used to design follow-up research projects. TRIAL REGISTRATION NUMBER: VfD_METH_MD_15_003600.

How socioeconomic inequalities impact pathways of care for coronary artery disease among elderly patients: study protocol for a qualitative longitudinal study.

INTRODUCTION: Several studies have identified that socioeconomic inequalities in coronary artery disease (CAD) morbidity and mortality lead to a disadvantage in patients with low socioeconomic status (SES). International studies have shown that socioeconomic inequalities also exist in terms of access, utilisation and quality of cardiac care. The aim of this qualitative study is to provide information on the impact of socioeconomic inequalities on the pathway of care for CAD, and to establish which factors lead to socioeconomic inequality of care to form and expand existing scientific theories. METHODS AND ANALYSIS: A longitudinal qualitative study with 48 patients with CAD, aged 60-80 years, is being conducted. Patients have been recruited consecutively at the University Hospital in Halle/Saale, Germany, and will be followed for a period of 6 months. Patients are interviewed two times face-to-face using semistructured interviews. Data are transcribed and analysed based on grounded theory. ETHICS AND DISSEMINATION: Only participants who have been informed and who have signed a declaration of consent have been included in the study. The study complies rigorously with data protection legislation. Approval of the Ethical Review Committee at the Martin-Luther University Halle-Wittenberg, Germany was obtained. The results of the study will be presented at several congresses, and will be published in high-quality peer-reviewed international journals. TRIAL REGISTRATION NUMBER: This study has been registered with the German Clinical Trials Register and assigned DRKS00007839.

Plant F-box protein evolution is determined by lineage-specific timing of major gene family expansion waves.

F-box proteins (FBPs) represent one of the largest and fastest evolving gene/protein families in the plant kingdom. The FBP superfamily can be divided in several subfamilies characterized by different C-terminal protein-protein interaction domains that recruit targets for proteasomal degradation. Hence, a clear picture of their phylogeny and molecular evolution is of special interest for the general understanding of evolutionary histories of multi-domain and/or large protein families in plants. In an effort to further understand the molecular evolution of F-box family proteins, we asked whether the largest subfamily in Arabidopsis thaliana, which carries a C-terminal F-box associated domain (FBA proteins) shares evolutionary patterns and signatures of selection with other FBPs. To address this question, we applied phylogenetic and molecular evolution analyses in combination with the evaluation of transcriptional profiles. Based on the 2219 FBA proteins we de novo identified in 34 completely sequenced plant genomes, we compared their evolutionary patterns to a previously analyzed large subfamily carrying C-terminal kelch repeats. We found that these two large FBP subfamilies generally tend to evolve by massive waves of duplication, followed by sequence conservation of the F-box domain and sequence diversification of the target recruiting domain. We conclude that the earlier in evolutionary time a major wave of expansion occurred, the more pronounced these selection signatures are. As a consequence, when performing cross species comparisons among FBP subfamilies, significant differences will be observed in the selective signatures of protein-protein interaction domains. Depending on the species, the investigated subfamilies comprise up to 45% of the complete superfamily, indicating that other subfamilies possibly follow similar modes of evolution.