Melanoma mutations modify melanocyte dynamics in co-culture with keratinocytes or fibroblasts.

Melanocytic cell interactions are integral to skin homeostasis, and affect the outcome of multiple diseases, including cutaneous pigmentation disorders and melanoma. By using automated-microscopy and machine-learning-assisted morphology analysis of primary human melanocytes in co-culture, we performed combinatorial interrogation of melanocyte genotypic variants and functional assessment of lentivirus-introduced mutations. Keratinocyte-induced melanocyte dendricity, an indicator of melanocyte differentiation, was reduced in the melanocortin 1 receptor (MC1R) R/R variant strain and by NRAS.Q61K and BRAF.V600E expression, while expression of CDK4.R24C and RAC1.P29S had no detectable effect. Time-lapse tracking of melanocytes in co-culture revealed dynamic interaction phenotypes and hyper-motile cell states that indicated that, in addition to the known role in activating mitogenic signalling, MEK-pathway-activating mutations may also allow melanocytes to escape keratinocyte control and increase their invasive potential. Expanding this combinatorial platform will identify other therapeutic target mutations and melanocyte genetic variants, as well as increase understanding of skin cell interactions.

Platelets kill circulating parasites of all major Plasmodium species in human malaria.

Platelets are understood to assist host innate immune responses against infection, although direct evidence of this function in any human disease, including malaria, is unknown. Here we characterized platelet-erythrocyte interactions by microscopy and flow cytometry in patients with malaria naturally infected with Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, or Plasmodium knowlesi Blood samples from 376 participants were collected from malaria-endemic areas of Papua, Indonesia, and Sabah, Malaysia. Platelets were observed binding directly with and killing intraerythrocytic parasites of each of the Plasmodium species studied, particularly mature stages, and was greatest in P vivax patients. Platelets preferentially bound to the infected more than to the uninfected erythrocytes in the bloodstream. Analysis of intraerythrocytic parasites indicated the frequent occurrence of platelet-associated parasite killing, characterized by the intraerythrocytic accumulation of platelet factor-4 and terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling of parasite nuclei (PF4+TUNEL+ parasites). These PF4+TUNEL+ parasites were not associated with measures of systemic platelet activation. Importantly, patient platelet counts, infected erythrocyte-platelet complexes, and platelet-associated parasite killing correlated inversely with patient parasite loads. These relationships, taken together with the frequency of platelet-associated parasite killing observed among the different patients and Plasmodium species, suggest that platelets may control the growth of between 5% and 60% of circulating parasites. Platelet-erythrocyte complexes made up a major proportion of the total platelet pool in patients with malaria and may therefore contribute considerably to malarial thrombocytopenia. Parasite killing was demonstrated to be platelet factor-4-mediated in P knowlesi culture. Collectively, our results indicate that platelets directly contribute to innate control of Plasmodium infection in human malaria.

Correction: Siegel et al. PEN-13: A New Generic 13-Item Questionnaire for Measuring Patient Enablement (German Version). Int. J. Environ. Res. Public Health 2019, 16, 4867.

In the original publication [...].

Addressing future work ability of employees in GP consultations: results of a cross-sectional study.

OBJECTIVES: In this study we examined to what extent members of a best-practice integrated healthcare model in Germany discussed their subjective future work ability with their general practitioner (GP); furthermore, we examined independent variables which explain whether future work ability is discussed. METHODS: In a cross-sectional survey, 1168 (out of 3218 invited) integrated healthcare members responded to a standardized questionnaire. This study includes n = 475 employed respondents who were at most 65 years old. We determined the (relative) frequency of employed members up to 65 years who had already discussed their subjective future work ability with their GP. By means of logistic regression analysis, explanatory variables were identified which statistically explained the discussion of future work ability with their GP. RESULTS: N = 80 (16.8%) respondents stated they had discussed their future work ability with their GP. A multiple logistic regression analysis showed the following results: The odds ratio for discussing future work ability is increased the more satisfied respondents are with their general practitioner, the worse they assess their current work ability in relation to the physical demands of the job, and when respondents suffer from one or more chronic diseases (Nagelkerke's pseudo-R2 = 0.13). CONCLUSIONS: Even in this healthcare setting, employees up to the age of 65 rarely discussed their subjective future work ability with their GP. This suggests that the issue 'future work ability' is even less commonly discussed in other community-based care settings in Germany. It seems that health care providers involved in acute care only sporadically take this issue into consideration - despite the great importance of maintaining work ability.

Work-Related Health Literacy: A Scoping Review to Clarify the Concept.

The overall aim of this scoping review is to outline the current state of research on health literacy (HL) in the workplace: the primary objective is to clarify the concept of individual work-related HL; the secondary aims are to report on interventions that promote individual work-related HL and to present respective measurement instruments validated to date. A high level of work-related HL could support work ability and in the longer run employability. These topics are becoming increasingly important in current circumstances and in view of ongoing developments (e.g., digitalization and "new" work). A basic understanding and measurement of HL as an individual competence in the context of working life is necessary to develop future interventions to promote HL among people of working age. According to the participants, concept, and context (PCC) framework, we included articles on health literacy (concept) in the target group of people of working age in the workplace (population and context). Key information sources were the databases PubMed, CINAHL, PsycInfo, and PSYNDEX. A total of 30 articles were included. There are several terms for "health literacy in the workplace" (including individual work-related or occupational HL). The conceptualizations of the individual employee's competence covered all aspects of HL ("access", "understand", "appraise", and "apply" health information). The conceptualizations differed, among others, in the covered time horizon (referring either only to employees' current work situation or additionally to their employability in the lifespan) or whether they referred also to the viability of the respective company. Published interventions attempting to promote individual work-related HL seem mostly to be targeted at the promotion of mental HL. A variety of outcomes have been measured in intervention studies, while specific measurement instruments for individual work-related HL seem to be scarce. We recommend the development of country-specific instruments for the assessment of individual work-related health literacy and to measure mental and physical work-related health literacy.

Attitudes of Company Executives toward a Comprehensive Workplace Health Management-Results of an Exploratory Cross-Sectional Study in Germany.

Workplace health management (WHM) in Germany aims at maintaining and increasing the health and well-being of employees. Little is known about company executives' attitudes toward WHM. To gain more insight, we conducted a large-scale survey in companies in the German county of Reutlingen in 2017. We sent a standardized questionnaire to 906 companies, containing inter alia 26 self-constructed declarative statements depicting company executives' opinions on various WHM aspects; 222 questionnaires could be evaluated. By exploratory factor analysis we assigned the 26 items to six factors reflecting different attitudes toward WHM. Factor values were standardized to a scale from 0 to 10. The attitude 'positive view of general health services in the company', for example, achieved by far the lowest mean agreement (3.3 points). For the attitude 'general skepticism toward WHM', agreement and disagreement were balanced (5.0 points). Using multiple regression analyses, we searched for variables that could partially explain respondents' agreement with attitudes. In conclusion, a general WHM skepticism was widespread, but not dominant. The idea that general health services should be offered in companies was predominantly rejected. Older respondents and respondents from smaller companies and craft enterprises were more skeptical than average about WHM and its possible extensions.

Dysregulated G2 phase checkpoint recovery pathway reduces DNA repair efficiency and increases chromosomal instability in a wide range of tumours.

Defective DNA repair is being demonstrated to be a useful target in cancer treatment. Currently, defective repair is identified by specific gene mutations, however defective repair is a common feature of cancers without these mutations. DNA damage triggers cell cycle checkpoints that are responsible for co-ordinating cell cycle arrest and DNA repair. Defects in checkpoint signalling components such as ataxia telangiectasia mutated (ATM) occur in a low proportion of cancers and are responsible for reduced DNA repair and increased genomic instability. Here we have investigated the AURKA-PLK1 cell cycle checkpoint recovery pathway that is responsible for exit from the G2 phase cell cycle checkpoint arrest. We demonstrate that dysregulation of PP6 and AURKA maintained elevated PLK1 activation to promote premature exit from only ATM, and not ATR-dependent checkpoint arrest. Surprisingly, depletion of the B55α subunit of PP2A that negatively regulates PLK1 was capable of overcoming ATM and ATR checkpoint arrests. Dysregulation of the checkpoint recovery pathway reduced S/G2 phase DNA repair efficiency and increased genomic instability. We found a strong correlation between dysregulation of the PP6-AURKA-PLK1-B55α checkpoint recovery pathway with signatures of defective homologous recombination and increased chromosomal instability in several cancer types. This work has identified an unrealised source of G2 phase DNA repair defects and chromosomal instability that are likely to be sensitive to treatments targeting defective repair.

Targeting Replication Stress Using CHK1 Inhibitor Promotes Innate and NKT Cell Immune Responses and Tumour Regression.

Drugs selectively targeting replication stress have demonstrated significant preclinical activity, but this has not yet translated into an effective clinical treatment. Here we report that targeting increased replication stress with a combination of Checkpoint kinase 1 inhibitor (CHK1i) with a subclinical dose of hydroxyurea targets also promotes pro-inflammatory cytokine/chemokine expression that is independent of cGAS-STING pathway activation and immunogenic cell death in human and murine melanoma cells. In vivo, this drug combination induces tumour regression which is dependent on an adaptive immune response. It increases cytotoxic CD8+ T cell activity, but the major adaptive immune response is a pronounced NKT cell tumour infiltration. Treatment also promotes an immunosuppressive tumour microenvironment through CD4+ Treg and FoxP3+ NKT cells. The number of these accumulated during treatment, the increase in FoxP3+ NKT cells numbers correlates with the decrease in activated NKT cells, suggesting they are a consequence of the conversion of effector to suppressive NKT cells. Whereas tumour infiltrating CD8+ T cell PD-1 and tumour PD-L1 expression was increased with treatment, peripheral CD4+ and CD8+ T cells retained strong anti-tumour activity. Despite increased CD8+ T cell PD-1, combination with anti-PD-1 did not improve response, indicating that immunosuppression from Tregs and FoxP3+ NKT cells are major contributors to the immunosuppressive tumour microenvironment. This demonstrates that therapies targeting replication stress can be well tolerated, not adversely affect immune responses, and trigger an effective anti-tumour immune response.

Validation of a German version of the Caregiver Quality of Life Index-Cancer (CQOLC) in a sample of significant others of breast and gynaecologic cancer patients.

There is no disease-specific instrument to measure the quality of life of significant others of cancer patients in Germany. In this study, we evaluated the reliability and construct validity of a German version of the Caregiver Quality of Life Index-Cancer (CQOLC) in a sample of 212 caregivers of breast and gynaecologic cancer patients. The CQOLC was administered along with the World Health Organization Quality of Life short version (WHOQOL-BREF) to caregivers of patients taking part in a randomized-controlled intervention study. Data of 212 caregivers were gained at the baseline of the study. Internal consistency was determined by Cronbach's α. Construct validity was examined by conducting a confirmatory factor analysis (CFA) and hypothesis testing. Correlations between change scores with patients' global health-related quality of life (HRQoL) were calculated for three time points to evaluate the responsiveness. The three subscales "burden", "disruptiveness", and "financial concerns" indicate to a good reliability of the instrument (Cronbach's α ranged between 0.754 and 0.832), while the subscale "positive adaptation" demonstrated low reliability (α = 0.579). A CFA based on data from the whole set of CQOLC items resulted in CFI levels < .90, and a CFA without problematic items resulted in CFI levels also < .90.The construct validity of the CQOLC could be approved by a moderate to high convergence with close variables as the global HRQoL. Mean differences between caregivers of curatively or palliatively treated patients were nonsignificant (p = 0.959) at T1. Correlations for responsiveness were low with correlation coefficients ranging from 0.030 to 0.326. These data indicate that additional research is needed to further verify the validity of the instrument. The German scale of the CQOLC might be appropriate for clinical and research use, if the wording of some items is refined and if content validity is also assessed by caregivers themselves. The assessment of cancer patients' caregiver's quality of life can contribute to a better understanding of the effects of patient-oriented interventions including also closely involved next of kin's around the cancer patients.

The Relationship between Health Literacy, Quality of Life, and Subjective Health: Results of a Cross-Sectional Study in a Rural Region in Germany.

Low health literacy is associated with poorer health and quality of life. An open question is whether a regional integrated healthcare system whose management strives to enable and empower its members with regard to health issues can contribute to a higher level of health literacy in the population. Against this background, in a cross-sectional study we surveyed a random selection of members of an integrated healthcare system in southwestern Germany (n = 180). The standardized questionnaire included, inter alia, questions on health literacy and subjective health. In this sample we wanted to (1) determine the proportion of respondents with sufficient health literacy and (2) investigate whether the association between health literacy and quality of life and subjective health status-as found in other studies-can be replicated. In our sample a health literacy score could be calculated in 126 subjects (70%). A sufficient level of health literacy was detected in 62% of respondents. Confirming the findings of a meta-analysis based on international studies, we found moderate correlations between health literacy and quality of life (r = 0.41) and health literacy and subjective health status (r = 0.40); these correlations hardly decreased when we controlled for various sociodemographic characteristics. As the proportion of respondents with sufficient health literacy was higher in our sample than in comparable studies conducted in Germany, we may hypothesize that an integrated healthcare system like the one we surveyed could have contributed to increased health literacy in the population. Thus, it could be worthwhile to investigate this research question with a more rigorous study design and a larger sample.

Host Porphobilinogen Deaminase Deficiency Confers Malaria Resistance in Plasmodium chabaudi but Not in Plasmodium berghei or Plasmodium falciparum During Intraerythrocytic Growth.

An important component in host resistance to malaria infection are inherited mutations that give rise to abnormalities and deficiencies in erythrocyte proteins and enzymes. Understanding how such mutations confer protection against the disease may be useful for developing new treatment strategies. A mouse ENU-induced mutagenesis screen for novel malaria resistance-conferring mutations identified a novel non-sense mutation in the gene encoding porphobilinogen deaminase (PBGD) in mice, denoted here as PbgdMRI58155. Heterozygote PbgdMRI58155 mice exhibited ~50% reduction in cellular PBGD activity in both mature erythrocytes and reticulocytes, although enzyme activity was ~10 times higher in reticulocytes than erythrocytes. When challenged with blood-stage P. chabaudi, which preferentially infects erythrocytes, heterozygote mice showed a modest but significant resistance to infection, including reduced parasite growth. A series of assays conducted to investigate the mechanism of resistance indicated that mutant erythrocyte invasion by P. chabaudi was normal, but that following intraerythrocytic establishment a significantly greater proportions of parasites died and therefore, affected their ability to propagate. The Plasmodium resistance phenotype was not recapitulated in Pbgd-deficient mice infected with P. berghei, which prefers reticulocytes, or when P. falciparum was cultured in erythrocytes from patients with acute intermittent porphyria (AIP), which had modest (20-50%) reduced levels of PBGD. Furthermore, the growth of Pbgd-null P. falciparum and Pbgd-null P. berghei parasites, which grew at the same rate as their wild-type counterparts in normal cells, were not affected by the PBGD-deficient background of the AIP erythrocytes or Pbgd-deficient mice. Our results confirm the dispensability of parasite PBGD for P. berghei infection and intraerythrocytic growth of P. falciparum, but for the first time identify a requirement for host erythrocyte PBGD by P. chabaudi during in vivo blood stage infection.

Caring for Workers' Health: Do German Employers Follow a Comprehensive Approach Similar to the Total Worker Health Concept? Results of a Survey in an Economically Powerful Region in Germany.

Similar to 'Total Worker Health' in the United States (USA), 'Workplace Health Management' in Germany is a holistic strategy to protect, promote, and manage employees' health at the workplace. It consists of four subcategories. While the subcategories 'occupational health and safety' and 'reintegration management' contain measures prescribed by law, 'workplace health promotion' and 'personnel development' can be designed more individually by the companies. The present study focused on the current implementation of voluntary and legally required measures of the four subcategories, as well as companies' satisfaction with the implementation. A total of N = 222/906 companies (small, medium, and big enterprises of one German county) answered a standardized questionnaire addressing the implementation of health-related measures, satisfaction with the implementation, and several company characteristics. In the subcategory 'occupational health and safety', 23.9% of the companies fulfilled all of the legally required measures, whereas in the category 'reintegration management', that rate amounted to 50.9%. There was a positive correlation between company size and the implementation grade, and as well between company size and the fulfilling of measures required by law. Companies tended to be more satisfied with higher implementation grades. Nevertheless, a surprisingly high proportion of the companies with poor implementation indicated satisfaction with the measures' implementation.

PEN-13: A New Generic 13-Item Questionnaire for Measuring Patient Enablement (German Version).

Background: The purpose of our study was to develop and psychometrically test a German-language survey instrument that measures patient enablement generically and in greater detail than previous instruments. Methods: A multidisciplinary team developed 13 items to capture individual aspects of patient enablement (PEN-13). A pre-test with 26 subjects was followed by a random sample survey of N = 1168 subjects. An exploratory factor analysis was conducted in a random split-half sample of the data to explore PEN-13's factor structure; a confirmatory factor analysis was conducted in the validation sample. The internal consistency of the factors was evaluated using Cronbach's alpha, PEN-13's construct validity was checked by means of additional hypothesis testing. Results: The two factors self-management and patient-practitioner interaction, detected in the exploratory analysis, were confirmed with a few modifications in the confirmatory factor analysis, with the comparative fit index (CFI) amounting to 0.903. The Cronbach's alpha values of those two factors amounted to α = 0.90 and α = 0.82, respectively. The correlations of the PEN-13 score with the 'general self-efficacy' and 'health literacy' (HLS-EU-Q16) scores further confirmed its construct validity; the respective correlation coefficients amounted to 0.57 and 0.60. Conclusion: The German version of the survey instrument Patient Enablement Scale-13 items (PEN-13) shows acceptable psychometric properties. Practical implications: PEN-13 seems particularly suitable for health services research purposes. We recommend checking the results in another sample as well as evaluating its responsiveness to enablement-enhancing interventions.

Defense Peptides Engineered from Human Platelet Factor 4 Kill Plasmodium by Selective Membrane Disruption.

Malaria is a serious threat to human health and additional classes of antimalarial drugs are greatly needed. The human defense protein, platelet factor 4 (PF4), has intrinsic antiplasmodial activity but also undesirable chemokine properties. We engineered a peptide containing the isolated PF4 antiplasmodial domain, which through cyclization, retained the critical structure of the parent protein. The peptide, cPF4PD, killed cultured blood-stage Plasmodium falciparum with low micromolar potency by specific disruption of the parasite digestive vacuole. Its mechanism of action involved selective penetration and accumulation inside the intraerythrocytic parasite without damaging the host cell or parasite membranes; it did not accumulate in uninfected cells. This selective activity was accounted for by observations of the peptide's specific binding and penetration of membranes with exposed negatively charged phospholipid headgroups. Our findings highlight the tremendous potential of the cPF4PD scaffold for developing antimalarial peptide drugs with a distinct and selective mechanism of action.

Validation of a German version of the Caregiver Quality of Life Index-Cancer (CQOLC) in a sample of significant others of breast and gynaecologic cancer patients

There is no disease-specific instrument to measure the quality of life of significant others of cancer patients in Germany. In this study, we evaluated the reliability and construct validity of a German version of the Caregiver Quality of Life Index-Cancer (CQOLC) in a sample of 212 caregivers of breast and gynaecologic cancer patients. The CQOLC was administered along with the World Health Organization Quality of Life short version (WHOQOL-BREF) to caregivers of patients taking part in a randomized-controlled intervention study. Data of 212 caregivers were gained at the baseline of the study. Internal consistency was determined by Cronbach's α. Construct validity was examined by conducting a confirmatory factor analysis (CFA) and hypothesis testing. Correlations between change scores with patients' global health-related quality of life (HRQoL) were calculated for three time points to evaluate the responsiveness. The three subscales "burden", "disruptiveness", and "financial concerns" indicate to a good reliability of the instrument (Cronbach's α ranged between 0.754 and 0.832), while the subscale "positive adaptation" demonstrated low reliability ( α = 0.579). A CFA based on data from the whole set of CQOLC items resulted in CFI levels < .90, and a CFA without problematic items resulted in CFI levels also < .90. The construct validity of the CQOLC could be approved by a moderate to high convergence with close variables as the global HRQoL. Mean differences between caregivers of curatively or palliatively treated patients were nonsignificant ( p = 0.959) at T1. Correlations for responsiveness were low with correlation coefficients ranging from 0.030 to 0.326. These data indicate that additional research is needed to further verify the validity of the instrument. The German scale of the CQOLC might be appropriate for clinical and research use, if the wording of some items is refined and if content validity is also assessed by caregivers themselves. The assessment of cancer patients' caregiver's quality of life can contribute to a better understanding of the effects of patient-oriented interventions including also closely involved next of kin's around the cancer patients.