Differentiation shifts from a reversible to an irreversible heterochromatin state at the DM1 locus.

Epigenetic defects caused by hereditary or de novo mutations are implicated in various human diseases. It remains uncertain whether correcting the underlying mutation can reverse these defects in patient cells. Here we show by the analysis of myotonic dystrophy type 1 (DM1)-related locus that in mutant human embryonic stem cells (hESCs), DNA methylation and H3K9me3 enrichments are completely abolished by repeat excision (CTG2000 expansion), whereas in patient myoblasts (CTG2600 expansion), repeat deletion fails to do so. This distinction between undifferentiated and differentiated cells arises during cell differentiation, and can be reversed by reprogramming of gene-edited myoblasts. We demonstrate that abnormal methylation in DM1 is distinctively maintained in the undifferentiated state by the activity of the de novo DNMTs (DNMT3b in tandem with DNMT3a). Overall, the findings highlight a crucial difference in heterochromatin maintenance between undifferentiated (sequence-dependent) and differentiated (sequence-independent) cells, thus underscoring the role of differentiation as a locking mechanism for repressive epigenetic modifications at the DM1 locus.

A review of the impact of maize-legume intercrops on the diversity and abundance of entomophagous and phytophagous insects.

In many parts of the world, chemical pesticides are the primary method of pest control in maize (Zea mays L.) crops. Concerns about the negative consequences of chemical pesticide use on people's health and the environment, as well as the emergence of insecticide resistance, have accelerated attempts to discover alternatives that are effective, low-risk, and cost-effective. Maize-legume intercropping systems are known to have multiple benefits to agroecosystem functioning, including pest regulation. This review focuses on the influence of maize-legume intercropping systems on insect diversity and abundance as a mechanism for insect pest regulation in maize crops. First, this review combines knowledge of maize-legume intercrops, with a particular emphasis on the mechanism by which this practice attracts beneficial insects (e.g., predators, parasitoids) to reduce pest damage in intercropping systems. In addition, the pairings of specific legume species with the greatest potential to attract more beneficial insects and therefore reduce maize pests are also discussed. Finally, future research needs are also recommended. Findings are reviewed in the context of looking for long-term management strategies that can increase the adoption of integrated pest management programs in maize-based production systems.

Physician-Specific Symptoms of Burnout Compared to a Non-Physicians Group.

Physician burnout is a systemic problem in health care due to its high prevalence and its negative impact on professional functioning and individual well-being. While unique aspects of the physician role contributing to the development burnout have been investigated recently, it is currently unclear whether burnout manifests differently in physicians compared to the non-physician working population. We conducted an individual symptom analysis of burnout symptoms comparing a large sample of physicians with a non-physician group. In this cross-sectional online study, burnout was assessed with the Maslach Burnout Inventory-General Survey. We matched physicians with non-physicians regarding their age, gender, educational level, occupational status, and total burnout level using a "nearest neighbour matching" procedure. We then conducted a series of between-groups comparisons. Data of 3846 (51.0% women) participants including 641 physicians and 3205 non-physicians were analysed. The most pronounced difference was that physicians were more satisfied with their work performance (medium effect size (r = 0.343). Our findings indicate minor yet significant differences in burnout phenomenology between physicians and non-physicians. This demonstrates unique aspects of physician burnout and implies that such differences should be considered in occupational research among physicians, particularly when developing burnout prevention programs for physicians.

A simple and low-cost resazurin assay for vitality assessment across species.

Working with biological organisms requires knowledge about the state of their viability and vitality to ascertain efficient processes. The phenoxazine dye resazurin is routinely used for viability assessment of many different species. Here, a novel use for resazurin as an indicator for vitality assessment across several species is proposed. Different amounts of biomass as well as mixtures of live/dead biomass were investigated for their capabilities of metabolizing resazurin and monitored over time. Increasing (live) biomass was found to increase reaction rate in a linear fashion, giving information about the cells' vitality. In an application example, stored suspension cultures of Sporosarcina pasteurii were found to decrease in viability over time, while urease activity decreased as well. For the first time, the assessment of vitality by one technique was demonstrated for several species in parallel.

Kidney Transplantation in a Recipient with Positive RT-PCR for SARS-CoV-2.

When it is safe to proceed with transplantation after coronavirus disease 2019 (COVID-19) infection is still unknown. We describe the clinical course and management of immunosuppression in a patient with positive real-time polymerase chain reaction (RT-PCR) for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) in a nasopharyngeal swab at the time of kidney transplantation, and with positive antibodies for SARS-CoV-2. The patient had no complications and was discharged with a functioning graft.

Relationship between job burnout and somatic diseases: a network analysis.

Job burnout, characterized by feelings of exhaustion, cynicism and reduced personal efficacy, has been defined as a risk state for the development of diseases, but its association with somatic diseases is underexplored. Study participants were 5671 respondents (mean age 44.1 years, range 18-70; 38.6% men) to an online survey of job burnout, using a mobile health web application. Respondents provided data on sociodemographic characteristics, symptoms of burnout, measured with the Maslach Burnout Inventory-General Survey, depressive symptoms, measured with the Profile of Mood States, and 11 categories of somatic diseases. Adjusting for age, sex, educational level, depressive symptoms, and all disease categories included, network analysis showed a significant association of high exhaustion with "high blood pressure" and a category of "other chronic somatic diseases". Low personal efficacy showed a significant association with "chronic lung diseases". In network analysis, clinically relevant depressive symptoms were also significantly associated with "high blood pressure", "other chronic somatic diseases" and "skin diseases". Logistic regression analysis confirmed these associations. Burnout dimensions were significantly associated with an increased risk for somatic diseases, independently of sociodemographic factors and clinically relevant depressive symptoms. This relationship seems particularly evident for the exhaustion dimension of job burnout.

[Conservative treatment of periprosthetic humeral fractures years after cemented fracture prostheses : A case series]. / Konservative Therapie periprothetischer Humerusfrakturen Jahre nach zementierter Frakturprothese : Eine Fallserie.

BACKGROUND: The exact incidence of postoperative periprosthetic humeral fractures (PPHF) months or years after fracture-related implantation of a hemiprosthesis is unknown. The currently available literature is predominantly concerned with operative treatment approaches. As a rule, these involved older patients and severe fracture conditions so that severe complications and unsatisfactory healing results were described. This article presents an alternative conservative treatment approach which is discussed based on the results of treatment. PATIENTS AND METHODS: Between 2011 and 2016 a conservative treatment of 5 female patients with PPHF could be carried out. Of the patients 4 were clinically and radiologically controlled at a mean follow-up time of 23 months. The fifth patient died 2 months after the trauma and only partial information of the treatment was available. RESULT: There were no intrahospital complications and just one posthospital complication. In the case of the patient who later died, repeated and unauthorized removal of the upper arm brace occurred in the nursing institution resulting in a lesion of the radial nerve. Of the four patients who completed treatment, three were very satisfied with the outcome of treatment. The mean DASH (Disabilities of Arm, Shoulder and Hand) and Oxford shoulder scores were on average 74 and 25 points, respectively. At the time of the follow-up examination all patients were free of pain, without the use of analgesics; however, there were still some limitations in the activities of daily life, which in three of the four patients was similar to the results following implantation of the fracture prosthesis. CONCLUSION: The conservative treatment of PPHF can be a safe treatment option in multimorbid and chronically ill patients. A close outpatient control and good patient compliance are important. In incompliant and dementia patients, the risk of failure of conservative treatment is increased.

Quantitative analysis of receptor-mediated uptake and pro-apoptotic activity of mistletoe lectin-1 by high content imaging.

Ribosome inactivating proteins (RIPs) are highly potent cytotoxins that have potential as anticancer therapeutics. Mistletoe lectin 1 (ML1) is a heterodimeric cytotoxic protein isolated from European Mistletoe and belongs to RIP class II. The aim of this project was to systematically study ML1 cell binding, endocytosis pathway(s), subcellular processing and apoptosis activation. For this purpose, state of the art cell imaging equipment and automated image analysis algorithms were used. ML1 displayed very fast binding to sugar residues on the membrane and energy-dependent uptake in CT26 cells. The co-staining with specific antibodies and uptake blocking experiments revealed involvement of both clathrin-dependent and -independent pathways in ML1 endocytosis. Co-localization studies demonstrated the toxin transport from early endocytic vesicles to Golgi network; a retrograde road to the endoplasmic reticulum. The pro-apoptotic and antiproliferative activity of ML1 were shown in time lapse movies and subsequently quantified. ML1 cytotoxicity was less affected in multidrug resistant tumor cell line 4T1 in contrast to commonly used chemotherapeutic drug (ML1 resistance index 6.9 vs 13.4 for doxorubicin; IC50: ML1 1.4 ng/ml vs doxorubicin 24000 ng/ml). This opens new opportunities for the use of ML1 as an alternative treatment in multidrug resistant cancers.

Pristinamycin-induced arthralgia and myalgia: Analysis of the French Pharmacovigilance Database.

INTRODUCTION: Pristinamycin is an antibiotic of the streptogramin family; few adverse effects of this drug are reported, only cutaneous and digestive ones. Arthralgia and myalgia may however be observed although not mentioned in the summary of product characteristics. OBJECTIVE: Description and analysis of cases of pristinamycin-induced arthralgia and/or myalgia registered in the French database of pharmacovigilance. METHOD: We carried out a targeted search of the database, selecting case patients presenting with arthralgia and muscle pain and excluding those associated with sensitivities or allergies to pristinamycin. RESULTS: We retrieved 15 case patients of pristinamycin-induced arthralgia and myalgia. Pristinamycin was the only potentially incriminated drug for seven case patients. CONCLUSION: Although not serious, this adverse effect deserves to be better known by physicians to optimize therapeutic management.

CRISPR/Cas9-Induced (CTG⋠CAG)n Repeat Instability in the Myotonic Dystrophy Type 1 Locus: Implications for Therapeutic Genome Editing.

Myotonic dystrophy type 1 (DM1) is caused by (CTG⋅CAG)n-repeat expansion within the DMPK gene and thought to be mediated by a toxic RNA gain of function. Current attempts to develop therapy for this disease mainly aim at destroying or blocking abnormal properties of mutant DMPK (CUG)n RNA. Here, we explored a DNA-directed strategy and demonstrate that single clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-cleavage in either its 5' or 3' unique flank promotes uncontrollable deletion of large segments from the expanded trinucleotide repeat, rather than formation of short indels usually seen after double-strand break repair. Complete and precise excision of the repeat tract from normal and large expanded DMPK alleles in myoblasts from unaffected individuals, DM1 patients, and a DM1 mouse model could be achieved at high frequency by dual CRISPR/Cas9-cleavage at either side of the (CTG⋅CAG)n sequence. Importantly, removal of the repeat appeared to have no detrimental effects on the expression of genes in the DM1 locus. Moreover, myogenic capacity, nucleocytoplasmic distribution, and abnormal RNP-binding behavior of transcripts from the edited DMPK gene were normalized. Dual sgRNA-guided excision of the (CTG⋅CAG)n tract by CRISPR/Cas9 technology is applicable for developing isogenic cell lines for research and may provide new therapeutic opportunities for patients with DM1.

Risk assessment for job burnout with a mobile health web application using questionnaire data: a proof of concept study.

BACKGROUND: Job burnout has become a rampant epidemic in working societies, causing high productivity loss and healthcare costs. An easy accessible tool to detect clinically relevant risk may bear the potential to timely avert the dire sequelae of burnout. As a start, we performed a proof of concept study to test the utilization of a mobile health web application for a free and anonymous burnout risk assessment with established questionnaires. METHODS: We designed a client-side javascript web application for users who filled out demographic and psychometric data forms over the internet. Users were recruited through social media, back links from hospital websites, and search engine optimization. Similar to population-based studies, we used the Maslach Burnout Inventory-General Survey (MBI-GS) to calculate a burnout risk index (BRIX). As additional mental health burden indices, users filled out the Perceived Stress Scale, Insomina Severity Index, and Profile of Mood States. RESULTS: Within six months, the MBI-GS was completed by 11,311 users (median age 33 years, 85 % women) of whom 20.0 % had no clinically relevant burnout risk, 54.7 % had mild-to-moderate risk, and 25.3 % had high risk. In the 2947 users completing all questionnaires, female sex (B = -0.03), cohabiting (B = -0.03), negative affect (B = 0.46), positive affect (B = -0.20), perceived stress (B = 0.18), and insomnia symptoms (B = 0.04) explained 56.2 % of the variance in the continuously scaled BRIX. The reliability was good to excellent for all psychometric scales. The weighting of the BRIX with mental health burden indices primarily modified the risk in users with mild-to-moderate burnout risk. CONCLUSIONS: A low-threshold web application can reliably assess the risk of job burnout. As the bulk of users had clinically relevant burnout scores, a web application may be useful to target employees at risk. The clinical value of the BRIX and its modification with coexistent/absent mental health burden awaits evaluation with work and health outcomes.

A low absolute number of expanded transcripts is involved in myotonic dystrophy type 1 manifestation in muscle.

Muscular manifestation of myotonic dystrophy type 1 (DM1), a common inheritable degenerative multisystem disorder, is mainly caused by expression of RNA from a (CTG·CAG)n-expanded DM1 locus. Here, we report on comparative profiling of expression of normal and expanded endogenous or transgenic transcripts in skeletal muscle cells and biopsies from DM1 mouse models and patients in order to help us in understanding the role of this RNA-mediated toxicity. In tissue of HSA(LR) mice, the most intensely used 'muscle-only' model in the DM1 field, RNA from the α-actin (CTG)250 transgene was at least 1000-fold more abundant than that from the Dmpk gene, or the DMPK gene in humans. Conversely, the DMPK transgene in another line, DM500/DMSXL mice, was expressed ∼10-fold lower than the endogenous gene. Temporal regulation of expanded RNA expression differed between models. Onset of expression occurred remarkably late in HSA(LR) myoblasts during in vitro myogenesis whereas Dmpk or DMPK (trans)genes were expressed throughout proliferation and differentiation phases. Importantly, quantification of absolute transcript numbers revealed that normal and expanded Dmpk/DMPK transcripts in mouse models and DM1 patients are low-abundance RNA species. Northern blotting, reverse transcriptase-quantitative polymerase chain reaction, RNA-sequencing and fluorescent in situ hybridization analyses showed that they occur at an absolute number between one and a few dozen molecules per cell. Our findings refine the current RNA dominance theory for DM1 pathophysiology, as anomalous factor binding to expanded transcripts and formation of soluble or insoluble ribonucleoprotein aggregates must be nucleated by only few expanded DMPK transcripts and therefore be a small numbers game.

Cell membrane integrity in myotonic dystrophy type 1: implications for therapy.

Myotonic Dystrophy type 1 (DM1) is a multisystemic disease caused by toxic RNA from a DMPK gene carrying an expanded (CTG•CAG)n repeat. Promising strategies for treatment of DM1 patients are currently being tested. These include antisense oligonucleotides and drugs for elimination of expanded RNA or prevention of aberrant binding to RNP proteins. A significant hurdle for preclinical development along these lines is efficient systemic delivery of compounds across endothelial and target cell membranes. It has been reported that DM1 patients show elevated levels of markers of muscle damage or loss of sarcolemmal integrity in their serum and that splicing of dystrophin, an essential protein for muscle membrane structure, is abnormal. Therefore, we studied cell membrane integrity in DM1 mouse models commonly used for preclinical testing. We found that membranes in skeletal muscle, heart and brain were impermeable to Evans Blue Dye. Creatine kinase levels in serum were similar to those in wild type mice and expression of dystrophin protein was unaffected. Also in patient muscle biopsies cell surface expression of dystrophin was normal and calcium-positive fibers, indicating elevated intracellular calcium levels, were only rarely seen. Combined, our findings indicate that cells in DM1 tissues do not display compromised membrane integrity. Hence, the cell membrane is a barrier that must be overcome in future work towards effective drug delivery in DM1 therapy.

[Multidisciplinary health care needs of psychologically distressed cancer patients in a Comprehensive Cancer Center]. / Multidisziplinärer Versorgungsbedarf psychisch belasteter Tumorpatienten an einem Comprehensive Cancer Center.

BACKGROUND AND OBJECTIVE: Considering the prolonged life-expectancies and the resulting demands that are placed on cancer patients and their relatives, the importance of specific counseling and support services including psycho-oncology, social services, nutritional, and exercise counseling has profoundly increased. The main focus of the current study was to evaluate the multidisciplinary health care needs of emotionally distressed cancer patients whoe were treated in a Comprehensive Cancer Center. METHODS AND STUDYGROUP: 831 out-patients were evaluated with regard to their psychological distress level and their multidisciplinary health care needs for specialist services of psycho-oncology, social services, nutritional, and exercise counseling using a tablet-PC assisted screening questionnaire. Separate analyses were completed for patients with and without psychological distress. RESULTS: One third of the screened patients showed clinically relevant psychological distress. Health care needs for all specialist services were significantly greater among these patients compared to patients without psychological distress (all p-values < 0.005). The higher needs were foremost presented by the number of needed specialist services (p < 0.001): two thirds of the psychologically distressed patients demonstrated, besides the need for a psycho-oncological service, a need for two or three further specialist services, whereas among patients without psychological distress more than 70% showed a need for at most one specialist service. CONCLUSION: Multidisciplinary health care needs of psychologically distressed cancer patients should be systematically addressed in a Comprehensive Cancer Center, and patients should be offered a coordinated and integrated health care program.

[Smoking cessation in patients with COPD]. / Tabakentwöhnung bei COPD.

Chronic obstructive pulmonary disease (COPD) is a leading cause of death worldwide. Cigarette smoking is the main cause of COPD. Quitting smoking is thus the most effective treatment strategy and central in COPD prevention. A number of guidelines on prevention, diagnosis, therapy and rehabilitation of COPD have been published. To help implementing and standardizing smoking cessation in COPD a guideline was published 2008 in Germany focusing on this complex issue. The present guideline is an update of the 2008 guideline and has a high grade of evidence (S3 according to the AWMF; Arbeitsgemeinschaft wissenschaftlicher medizinischer Fachgesellschaften). The guideline gives comprehensive and practical information on how to integrate smoking cessation as an central part of COPD therapy.

How citrullination invaded rheumatoid arthritis research.

Citrullination and the immune response to citrullinated proteins have been fundamental for the early recognition of rheumatoid arthritis by serological tests and a better understanding of its pathophysiology. In the first years after the initial publications, the focus was on the antibodies directed to citrullinated proteins. It is now realized that citrullinating enzymes and citrullinated proteins may have important roles in the maintenance of the inflammatory processes in the joints. There is also accumulating evidence for a direct role of citrullination in tissue destruction in the rheumatoid synovium. Here we will discuss the development and importance of anti-citrullinated protein antibodies in rheumatoid arthritis as well as recent findings implicating citrullination in the pathophysiology of rheumatoid arthritis.

[Photodynamic therapy of bladder cancer. A new option]. / Die photodynamische Therapie des Harnblasenkarzinoms. Eine neue Option.

BACKGROUND: Bladder cancer (BCa) is the third most common tumor in Germany. Currently, resection therapy for superficial BCa (Ta, CIS) includes photodynamic diagnostics (PDD) using HEXVIX® for improved assessment of tumor spread. Trials using these photosensitizers for photodynamic therapy (PDT) showed only limited success. Especially low tissue penetration due to short-wave excitation was a limiting factor. METHODS: This study which was funded by the German Research Foundation (DFG) examined the feasibility of the novel photosensitizer tetrahydroporphyrin-tetratosylate (THPTS) for PDT in a rat bladder cancer model. RESULTS: As THPTS is very effectively excitable at a near infrared wavelength of 760 nm it is within the so-called phototherapeutic window and allows tissue penetration of up to 15 mm. Thus THPTS can also be used for PDT of larger, solid tumors as was previously demonstrated for other tumor entities. Therefore, effective treatment of even muscle-invasive bladder cancer (≥T2) may become an option using THPTS. In this current study the effectiveness and safety of THPTS-PDT was examined in an orthotopic bladder cancer rat model.