Automated manufacture of ΔNPM1 TCR-engineered T cells for AML therapy.

Acute myeloid leukemia (AML) is a heterogeneous malignancy that requires further therapeutic improvement, especially for the elderly and for subgroups with poor prognosis. A recently discovered T cell receptor (TCR) targeting mutant nucleophosmin 1 (ΔNPM1) presents an attractive option for the development of a cancer antigen-targeted cellular therapy. Manufacturing of TCR-modified T cells, however, is still limited by a complex, time-consuming, and laborious procedure. Therefore, this study specifically addressed the requirements for a scaled manufacture of ΔNPM1-specific T cells in an automated, closed, and good manufacturing practice-compliant process. Starting from cryopreserved leukapheresis, 2E8 CD8-positive T cells were enriched, activated, lentivirally transduced, expanded, and finally formulated. By adjusting and optimizing culture conditions, we additionally reduced the manufacturing time from 12 to 8 days while still achieving a clinically relevant yield of up to 5.5E9 ΔNPM1 TCR-engineered T cells. The cellular product mainly consisted of highly viable CD8-positive T cells with an early memory phenotype. ΔNPM1 TCR CD8 T cells manufactured with the optimized process showed specific killing of AML in vitro and in vivo. The process has been implemented in an upcoming phase 1/2 clinical trial for the treatment of NPM1-mutated AML.

Individual and environmental determinants of serum ferritin levels: A structural equation model.

BACKGROUND AND OBJECTIVES: Serum ferritin levels are increasingly being used to assess iron stores. Considerable variation in ferritin levels within and between individuals has been observed, but our current understanding of factors that explain this variation is far from complete. We aim to combine multiple potential determinants in an integrative model, and investigate their relative importance and potential interactions. METHODS: We use ferritin measurements collected by Sanquin Blood Bank on both prospective (N = 59 596) and active blood donors (N = 78 318) to fit a structural equation model with three latent constructs (individual characteristics, donation history, and environmental factors). Parameters were estimated separately by sex and donor status. RESULTS: The model explained 25% of ferritin variance in prospective donors, and 40% in active donors. Individual characteristics and donation history were the most important determinants of ferritin levels in active donors. The association between environmental factors and ferritin was smaller but still substantial; higher exposure to air pollution was associated with higher ferritin levels, and this association was considerably stronger for active blood donors than for prospective donors. DISCUSSION: In active donors, individual characteristics explain 20% (17%) of ferritin variation, donation history explains 14% (25%) and environmental factors explain 5% (4%) for women (men). Our model presents known ferritin determinants in a broader perspective, allowing for comparison with other determinants as well as between new and active donors, or between men and women.

Identification of patient-specific CD4+ and CD8+ T cell neoantigens through HLA-unbiased genetic screens.

Cancer neoantigens that arise from tumor mutations are drivers of tumor-specific T cell responses, but identification of T cell-recognized neoantigens in individual patients is challenging. Previous methods have restricted antigen discovery to selected HLA alleles, thereby limiting the breadth of neoantigen repertoires that can be uncovered. Here, we develop a genetic neoantigen screening system that allows sensitive identification of CD4+ and CD8+ T cell-recognized neoantigens across patients' complete HLA genotypes.

Mortality among rough sleepers, squatters, residents of homeless shelters or hotels and sofa-surfers: a pooled analysis of UK birth cohorts.

BACKGROUND: Homelessness encompasses a wide spectrum of experience. Rough sleepers and people attending homeless shelters have been found to be at an increased risk of mortality. It is unclear whether risks are also elevated in those squatting, living temporarily in low-cost hotels or 'sofa-surfing' with friends or family members. This study examines mortality in a representative nationwide sample of people who have slept rough, squatted, lived in shelters or low-cost hotels and sofa-surfed. METHODS: Using unpublished data from two national birth cohorts, namely the National Child Development Study and the 1970 British Birth Cohort study, Cox proportional-hazards models and random-effects meta-analyses were used to analyse associations between homelessness and different types of homeless experience (rough sleeping, squatting, staying in a homeless shelter or low-cost hotel, and sofa-surfing) and mortality. RESULTS: Out of the 23 678 participants, 1444 (6.1%) reported having been homeless and 805 (3.4%) deaths occurred. Homelessness was associated with an increased risk of mortality [hazard ratio (HR) 1.68, 95% confidence interval (CI) 1.24-2.26]. Mortality risk was raised across the spectrum of homeless experience, from sleeping rough (HR 4.71, 95% CI 2.38-9.30), to squatting (HR 6.35, 95% CI 2.73-14.75), staying in a homeless shelter (HR 4.89, 95% CI 2.36-10.11), staying in a low-cost hotel (HR 3.38, 95% CI 1.30-8.79 through to sofa-surfing (HR 2.86, 95% CI 1.84-4.42). Associations remained after separate control for socio-economic status, mental health, substance use, accidents and assaults, and criminality. CONCLUSIONS: Mortality rates were raised across all types of homeless experience. This included squatting and sofa-surfing that have not previously been reported. Studies that have omitted the less severe, but more prevalent, use of low-cost hotels and sofa-surfing may have underestimated the impacts of homelessness on mortality.

Donation-induced iron depletion is significantly associated with low hemoglobin at subsequent donations.

BACKGROUND: Blood donation is associated with a loss of hemoglobin (Hb)-bound iron. Hb levels recover relatively fast by using stored iron. However, it takes more time to replenish iron stores, potentially resulting in iron deficiency. STUDY DESIGN: Hb and ferritin levels were measured in 5056 new, first-time, and repeat whole blood donors. We investigated whether increasing numbers of donations are associated with lower ferritin levels. Furthermore, we tested whether low ferritin levels are associated with low-Hb deferral at the subsequent donation attempt by performing logistic regression adjusted for age and stratified by sex. RESULTS: Whereas mean Hb levels are relatively stable, ferritin levels significantly decrease with increasing numbers of donations and were approximately 50% lower for donors with >50 donations compared with those with 2-10 donations. Despite the poor correlation of ferritin and Hb levels, cross-sectional, iron-deficient donors (ferritin <15 ng/ml) had 21.8 (8.5-55.6) higher odds in men, 10.1 (6.1-16.5) in premenopausal women, and 11.7 (5.2-26.4) in postmenopausal women for Hb deferral at a subsequent visit. DISCUSSION: To conclude, repeated donations may induce iron deficiency, which corresponds with an over tenfold increased risk of having insufficiently restored Hb levels at a subsequent donation attempt. Longer donation intervals and/or higher dietary or supplemental iron intake are warranted to prevent accumulated iron depletion and subsequent low-Hb deferral in whole blood donors.

T cells expanded from renal cell carcinoma display tumor-specific CD137 expression but lack significant IFN-γ, TNF-α or IL-2 production.

Metastatic renal cell carcinoma (RCC) has a poor prognosis. Recent advances have shown beneficial responses to immune checkpoint inhibitors, such as anti-PD-1/PD-L1 antibodies. As only a subset of RCC patients respond, alternative strategies should be explored. Patients refractory to anti-PD-1 therapy may benefit from autologous tumor-infiltrating lymphocyte (TIL) therapy. Even though efficient TIL expansion was reported from RCC lesions, it is not well established how many RCC TIL products are tumor-reactive, how well they produce pro-inflammatory cytokines in response to autologous tumors, and whether their response correlates with the presence of specific immune cells in the tumor lesions. We here compared the immune infiltrate composition of RCC lesions with that of autologous kidney tissue of 18 RCC patients. Tcell infiltrates were increased in the tumor lesions, and CD8+ Tcell infiltrates were primarily of effector memory phenotype. Nine out of 16 (56%) tested TIL products we generated were tumor-reactive, as defined by CD137 upregulation after exposure to autologous tumor digest. Tumor reactivity was found in particular in TIL products originating from tumors with ahigh percentage of infiltrated Tcells compared to autologous kidney, and increased CD25 expression on CD8+ Tcells. Importantly, although TIL products had the capacity to produce the key effector cytokines IFN-γ, TNF-α or IL-2, they failed to produce significant amounts in response to autologous tumor digests. In conclusion, TIL products from RCC lesions contain tumor-reactive Tcells. Their restricted tumor-specific cytokine production requires further investigation of immunosuppressive factors in RCC and subsequent optimization of RCC-derived TIL culture conditions.

Stroke in COVID-19: A systematic review and meta-analysis.

BACKGROUND: Coronavirus disease 2019 (COVID-19) has become a global pandemic, affecting millions of people. However, the relationship between COVID-19 and acute cerebrovascular diseases is unclear. AIMS: We aimed to characterize the incidence, risk factors, clinical-radiological manifestations, and outcome of COVID-19-associated stroke. METHODS: Three medical databases were systematically reviewed for published articles on acute cerebrovascular diseases in COVID-19 (December 2019-September 2020). The review protocol was previously registered (PROSPERO ID = CRD42020185476). Data were extracted from articles reporting ≥5 stroke cases in COVID-19. We complied with the PRISMA guidelines and used the Newcastle-Ottawa Scale to assess data quality. Data were pooled using a random-effect model. SUMMARY OF REVIEW: Of 2277 initially identified articles, 61 (2.7%) were entered in the meta-analysis. Out of 108,571 patients with COVID-19, acute CVD occurred in 1.4% (95%CI: 1.0-1.9). The most common manifestation was acute ischemic stroke (87.4%); intracerebral hemorrhage was less common (11.6%). Patients with COVID-19 developing acute cerebrovascular diseases, compared to those who did not, were older (pooled median difference = 4.8 years; 95%CI: 1.7-22.4), more likely to have hypertension (OR = 7.35; 95%CI: 1.94-27.87), diabetes mellitus (OR = 5.56; 95%CI: 3.34-9.24), coronary artery disease (OR = 3.12; 95%CI: 1.61-6.02), and severe infection (OR = 5.10; 95%CI: 2.72-9.54). Compared to individuals who experienced a stroke without the infection, patients with COVID-19 and stroke were younger (pooled median difference = -6.0 years; 95%CI: -12.3 to -1.4), had higher NIHSS (pooled median difference = 5; 95%CI: 3-9), higher frequency of large vessel occlusion (OR = 2.73; 95%CI: 1.63-4.57), and higher in-hospital mortality rate (OR = 5.21; 95%CI: 3.43-7.90). CONCLUSIONS: Acute cerebrovascular diseases are not uncommon in patients with COVID-19, especially in those whom are severely infected and have pre-existing vascular risk factors. The pattern of large vessel occlusion and multi-territory infarcts suggests that cerebral thrombosis and/or thromboembolism could be possible causative pathways for the disease.

Dietary intake of heme iron is associated with ferritin and hemoglobin levels in Dutch blood donors: results from Donor InSight.

Whole blood donors, especially frequently donating donors, have a risk of iron deficiency and low hemoglobin levels, which may affect their health and eligibility to donate. Lifestyle behaviors, such as dietary iron intake and physical activity, may influence iron stores and thereby hemoglobin levels. We aimed to investigate whether dietary iron intake and questionnaire-based moderate-to-vigorous physical activity were associated with hemoglobin levels, and whether ferritin levels mediated these associations. In Donor InSight-III, a Dutch cohort study of blood and plasma donors, data on heme and non-heme iron intake (mg/day), moderate-to-vigorous physical activity (10 minutes/day), hemoglobin levels (mmol/L) and ferritin levels (µg/L) were available in 2,323 donors (1,074 male). Donors with higher heme iron intakes (regression coefficients (ß) in men and women: 0.160 and 0.065 mmol/L higher hemoglobin per 1 mg of heme iron, respectively) and lower non-heme iron intakes (ß: -0.014 and -0.017, respectively) had higher hemoglobin levels, adjusted for relevant confounders. Ferritin levels mediated these associations (indirect effect (95% confidence interval) in men and women respectively: 0.074 (0.045; 0.111) and 0.061 (0.030; 0.096) for heme and -0.003 (-0.008;0.001) and -0.008 (-0.013;-0.003) for non-heme). Moderate-to-vigorous physical activity was negatively associated with hemoglobin levels in men only (ß: -0.005), but not mediated by ferritin levels. In conclusion, higher heme and lower non-heme iron intake were associated with higher hemoglobin levels in donors, via higher ferritin levels. This indicates that donors with high heme iron intake may be more capable of maintaining iron stores to recover hemoglobin levels after blood donation.

Challenges in Establishing Pure Lung Cancer Organoids Limit Their Utility for Personalized Medicine.

Clinical implementation of tumor organoids for personalized medicine requires that pure tumor organoids can be reliably established. Here, we present our experience with organoid cultures from >70 non-small cell lung cancer (NSCLC) samples. We systematically evaluate several methods to identify tumor purity of organoids established from intrapulmonary tumors. Eighty percent of organoids from intrapulmonary lesions have a normal copy number profile, suggesting overgrowth by normal airway organoids (AOs). This is further supported by the failure to detect mutations found in the original tumor in organoids. Histomorphology alone is insufficient to determine tumor purity, but when combined with p63 immunostaining, tumor and normal AOs can be distinguished. Taking into account overgrowth by normal AOs, the establishment rate of pure NSCLC organoids is 17%. Therefore, current methods are insufficient to establish pure NSCLC organoids from intrapulmonary lesions. We discourage their use unless steps are taken to prevent overgrowth by normal AOs.

Lifestyle behaviours are not associated with haemolysis: results from Donor InSight.

BACKGROUND: Lifestyle behaviours such as physical activity, sedentary behaviour and dietary habits have been shown to influence blood lipid levels, and both lifestyle and blood lipids may be associated with haemolysis during storage of blood products. We aimed to investigate whether lifestyle behaviours are associated with degree of haemolysis in red cell concentrates (RCC), and if such associations are mediated by low-density lipoprotein (LDL) cholesterol and triglyceride levels. MATERIALS AND METHODS: Cross-sectional analyses were performed in data from 760 Dutch blood donors participating in Donor InSight, an observational cohort study. Linear regression analyses were conducted to assess associations of lifestyle behaviours with haemolysis levels in RCC 28 days after blood sampling. Lifestyle behaviours included moderate-to-vigorous physical activity and sedentary behaviour measured by accelerometry, and self-reported intake of a selection of foods potentially related to blood lipids, i.e. consumption of eggs, meat, nuts and fish. Potential mediating roles of both LDL cholesterol and triglyceride levels were investigated separately. All analyses were adjusted for relevant confounders. RESULTS: No statistically significant nor substantial associations of any of the lifestyle behaviours with haemolysis in RCC were found, nor were there any associations between lifestyle behaviours and blood lipids. We did find consistent positive associations of LDL cholesterol and triglyceride levels with haemolysis in RCC during storage. DISCUSSION: In this large cohort, blood lipid levels were consistently associated with haemolysis in RCC. Nonetheless, there was no evidence for an association between lifestyle behaviours and haemolysis in RCC, or for mediating effects by blood lipid levels.

Polyfunctional tumor-reactive T cells are effectively expanded from non-small cell lung cancers, and correlate with an immune-engaged T cell profile.

Non-small cell lung cancer (NSCLC) is the second most prevalent type of cancer. With the current treatment regimens, the mortality rate remains high. Therefore, better therapeutic approaches are necessary. NSCLCs generally possess many genetic mutations and are well infiltrated by T cells (TIL), making TIL therapy an attractive option. Here we show that T cells from treatment naive, stage I-IVa NSCLC tumors can effectively be isolated and expanded, with similar efficiency as from normal lung tissue. Importantly, 76% (13/17) of tested TIL products isolated from NSCLC lesions exhibited clear reactivity against primary tumor digests, with 0.5%-30% of T cells producing the inflammatory cytokine Interferon (IFN)-γ. Both CD4+ and CD8+ T cells displayed tumor reactivity. The cytokine production correlated well with CD137 and CD40L expression. Furthermore, almost half (7/17) of the TIL products contained polyfunctional T cells that produced Tumor Necrosis Factor (TNF)-α and/or IL-2 in addition to IFN-γ, a hallmark of effective immune responses. Tumor-reactivity in the TIL products correlated with high percentages of CD103+CD69+CD8+ T cell infiltrates in the tumor lesions, with PD-1hiCD4+ T cells, and with FoxP3+CD25+CD4+ regulatory T cell infiltrates, suggesting that the composition of T cell infiltrates may predict the level of tumor reactivity. In conclusion, the effective generation of tumor-reactive and polyfunctional TIL products implies that TIL therapy will be a successful treatment regimen for NSCLC patients.

The association between population density and blood lipid levels in Dutch blood donors.

BACKGROUND: In low and middle-income countries (LMIC), the total and LDL cholesterol and triglyceride levels of residents of urban areas are reported to be higher than those of rural areas. This may be due to differences in lifestyle behaviors between residents of urban areas and rural areas in LMIC. In this study, our aims were to (1) examine whether or not LDL cholesterol, total/HDL ratios and triglyceride levels of individuals in densely populated areas are higher than those of individuals living in less-densely populated areas in a high-income country (HIC) and (2) investigate the potential mediating roles of physical activity and sedentary behavior. METHODS: We used cross-sectional data from 2547 Dutch blood donors that participated in Donor InSight-III. Linear regression was used to analyze the association between population density and LDL cholesterol, total/HDL cholesterol ratio and HDL cholesterol. The mediating roles of moderate-to-vigorous physical activity (MVPA) and sedentary behavior were investigated in a subsample (n = 740) for which objectively measured MVPA/sedentary behavior data was available. Multiple mediation with linear regression analyses were performed and the product-of-coefficients method was used to calculate direct and indirect effects. RESULTS: Mean LDL cholesterol and median total cholesterol/HDL cholesterol ratio and triglyceride levels were 2.89, 3.43 and 1.29 mmol/L, respectively. Population density was not associated with LDL cholesterol [ß 0.00 (- 0.01; 0.01)], log transformed total/HDL cholesterol ratio [ß 1.00 (1.00; 1.00)] and triglyceride levels [ß 1.00 (0.99; 1.00)]. No statistically significant direct or indirect effects were found. CONCLUSION: Contrary to previous findings in LMIC, no evidence was found that population density is associated with blood lipid levels in blood donors in the Netherlands or that MVPA and sedentary behavior mediate this association. This may be the result of socioeconomic differences and, in part, may be due to the good health of the study population and the relatively high population density in the Netherlands. Also, compared to LMIC, differences in physical activity levels in more versus less populated areas may be less pronounced in HIC.

Urban-rural differences in the association between blood lipids and characteristics of the built environment: a systematic review and meta-analysis.

INTRODUCTION: The built environment defines opportunities for healthy eating and physical activity and may thus be related to blood lipids. The aim of this study is to systematically analyse the scientific evidence on associations between built-environment characteristics and blood lipid levels in adults. METHODS: PubMed, EMBASE and Web of Science were searched for peer-reviewed papers on population-based studies up to 9 October 2017. We included studies that reported on built-environment characteristics and blood lipid levels in adult populations (≥18 years). Two reviewers independently screened titles/abstracts and full-texts of papers and appraised the risk of bias of included studies using an adapted version of the Quality Assessment Tool for Quantitative Studies. We performed meta-analyses when five or more studies had sufficient homogeneity in determinant and outcome. RESULTS: After screening 6902 titles/abstracts and 141 potentially relevant full-text articles, we included 50 studies. Forty-seven studies explored associations between urban versus rural areas with blood lipid levels. Meta-analyses on urban versus rural areas included 133 966 subjects from 36 studies in total. Total cholesterol levels were significantly and consistently higher in urban areas as compared with rural areas (mean difference 0.37 mmol/L, 95% CI 0.27 to 0.48). Urban/rural differences in high density lipoprotein cholesterol were inconsistent across studies and the pooled estimate showed no difference (0.00 mmol/L 95% CI -0.03 to 0.04). Low density lipoprotein (LDL) cholesterol and triglyceride levels were higher in urban than in rural areas (mean difference 0.28, 95% CI 0.17 to 0.39 and 0.09, 95% CI 0.03 to 0.14, respectively). CONCLUSIONS: Total and LDL cholesterol levels and triglycerides were consistently higher in residents of urban areas than those of rural areas. These results indicate that residents of urban areas generally have less favourable lipid profiles as compared with residents of rural areas. PROSPERO REGISTRATION NUMBER: CRD42016043226.

Dutch midwives' views on and experiences with woman-centred care - A Q-methodology study.

BACKGROUND: Woman-centred care is a philosophy for midwifery care management of the childbearing woman. There is no mutually recognised internalised way in midwifery to provide woman-centred care. OBJECTIVE: To reveal midwives' distinct perspectives about woman-centred care. METHODS: A Q-methodology study amongst 48 Dutch community-based midwives who rank-ordered 39 statements on woman-centred care, followed by semi-structured interviews to motivate their ranking. By-person factor analysis was used to derive latent views, representing midwives (factors) with similar attitudes towards woman-centred care. The qualitative data was used to aid interpretation of the factors. RESULTS: Four distinct factors emerged: (1) the humane midwife, containing two twinning factors: (1+) The philosophical midwife, who is the woman's companion during childbearing in being an authentic individual human being; (1-) the human-rights midwife, who is the woman's advocate for achieving autonomy and self-determination regarding care during the childbearing period. (2) The quality-of-care midwife, who regards good perinatal health outcomes, responsive care and positive maternal experiences as benchmarks for the quality of woman-centred care. (3) The job-crafting midwife, who focuses on self-organisation while seeking balance between the childbearing woman, herself as a professional and an individual and as a colleague. CONCLUSION/IMPLICATIONS: Each factor represented specific perspectives feeding into woman-centred practice. Although the humane midwife seems to represent the dominant and preferable perspective of woman-centred care, awareness and exploration of and reflection on the thoughts patterns represented by the four different perspectives, should be considered in education and professional development of (student)midwives of be(com)ing a woman-centred midwife.

What factors explain socioeconomic inequalities in adults' television-related sitting time?

BACKGROUND: There are considerable socioeconomic inequalities in television-related sitting time, but there is little evidence for the explanatory mechanisms. We used a cohort of Belgian adults (25-60 years) and older adults (≥65 years) to examine the social cognitive, home environmental and health-related factors contributing to socioeconomic differences in television-related sitting. METHODS: We included 301 adults and 258 older adults (total n = 559). Linear regression analyses were used to examine the associations of education and occupational status with television-related sitting time, adjusted for age and gender. We assessed the explanatory power of social cognitive, home environmental and health-related factors using the traditional 'change-in-estimation method'. RESULTS: Those with low and medium education, respectively, engaged in 54 and 28 minutes per day more television-related sitting time than those with high education. We found no association between occupational status and television-related sitting time. Social cognitive factors explained 54% of the difference in television-related sitting time between those with low and high education, while home environmental factors only explained 6%, and health-related variables explained 10% of these differences. CONCLUSION: We found no occupational inequalities in television-related sitting time. Social cognitive variables such as attitude and modelling of the partner explained a large part of the educational inequalities in television-related sitting time. If confirmed by future studies, a focus on social cognition may help reduce sedentary behaviours in low-educated adults and diminish inequalities in sedentary behaviours.

Donor InSight: characteristics and representativeness of a Dutch cohort study on blood and plasma donors.

BACKGROUND AND OBJECTIVES: More insight into donor health and behaviour may contribute to more efficient and focused strategies regarding donor care and management. Donor InSight (DIS) is a Dutch cohort study of blood and plasma donors. We aimed to outline the objectives and methods of DIS, describe the cohort, and compare it to the active Dutch donor population. MATERIALS AND METHODS: In 2007-2009 (DIS-I, n = 31 338) and 2012-2013 (DIS-II, 34 826, of whom 22 132 also participated in DIS-I) questionnaire data on demographics, donation, lifestyle, family composition, health and disease were collected. A second follow-up (DIS-III, n = 3046), including donors with differing haemoglobin trajectories, was completed in 2015-2016. DIS-III includes data on genetic determinants, iron and red cell indices. Representativeness of the DIS-I sample for the entire Dutch donor population was assessed by comparing characteristics of both. RESULTS: Donor InSight was initially set up because of a need for more detailed information and evidence as a basis for decision-making in blood banks. DIS-I sample is comparable to the total Dutch donor population in terms of age, body mass index, haemoglobin level, blood pressure, blood type and donation behaviour. CONCLUSION: Donor InSight is a cohort study representative of the Dutch donor population. It provides evidence to support evidence-based decision making.

Generation of Tumor-Reactive T Cells by Co-culture of Peripheral Blood Lymphocytes and Tumor Organoids.

Cancer immunotherapies have shown substantial clinical activity for a subset of patients with epithelial cancers. Still, technological platforms to study cancer T-cell interactions for individual patients and understand determinants of responsiveness are presently lacking. Here, we establish and validate a platform to induce and analyze tumor-specific T cell responses to epithelial cancers in a personalized manner. We demonstrate that co-cultures of autologous tumor organoids and peripheral blood lymphocytes can be used to enrich tumor-reactive T cells from peripheral blood of patients with mismatch repair-deficient colorectal cancer and non-small-cell lung cancer. Furthermore, we demonstrate that these T cells can be used to assess the efficiency of killing of matched tumor organoids. This platform provides an unbiased strategy for the isolation of tumor-reactive T cells and provides a means by which to assess the sensitivity of tumor cells to T cell-mediated attack at the level of the individual patient.

Exploring the cross-sectional association between outdoor recreational facilities and leisure-time physical activity: the role of usage and residential self-selection.

BACKGROUND: The availability of outdoor recreational facilities is associated with increased leisure-time physical activity (PA). We investigated how much of this association is attributable to selection effects, and explored whether usage of recreational facilities was an explanatory mechanism. METHODS: We analysed data from 5199 participants in the SPOTLIGHT survey residing in five European urban regions. Adults completed a survey and a Google Street View-based virtual audit was conducted to objectively measure the availability of outdoor recreational facilities in the residential neighbourhood. We used negative binomial GEE models to examine the association between objective and subjective availability of outdoor recreational facilities and leisure-time PA, and explored whether this association was attenuated after adjustment for socioeconomic status and preference for neighbourhoods with recreational facilities (as indicators of self-selection). We examined whether reported use of recreational facilities was associated with leisure-time PA (as explanatory mechanism), and summarized the most important motivations for (not) using recreational facilities. RESULTS: Subjective - but not objective - availability of outdoor recreational facilities was associated with higher levels of total leisure-time PA. After adjustment for self-selection (which attenuated the association by 25%), we found a 25% difference in weekly minutes of total leisure-time PA between individuals with and without self-reported availability of outdoor recreational facilities. For our study population, this translates to about 28 min per week. Participants who reported outdoor recreational facilities to be present but indicated not to use them (RR = 1.19, 95% CI = 1.03;1.22), and those reporting outdoor recreational facilities to be present and to use them (RR = 1.33, 95% CI = 1.22, 1.45) had higher levels of total leisure-time PA than those who reported outdoor recreational facilities not to be present. Proximity to outdoor recreational facilities was the most important motivation for use. CONCLUSION: The modest attenuation in the association between availability of outdoor recreational facilities and self-reported leisure-time PA suggests that individuals' higher activity levels may be due more to the perceived availability of outdoor recreational facilities than to self-selection. The use of these facilities seemed to be an important underlying mechanism, and proximity was the main motivator for using recreational facilities.

Woman-centered care 2.0: Bringing the concept into focus.

INTRODUCTION: Woman-centered care has become a midwifery concept with implied meaning. In this paper we aim to provide a clear conceptual foundation of woman-centered care for midwifery science and practice. METHODS: An advanced concept analysis was undertaken. At the outset, a systematic search of the literature was conducted in PubMed, OVID and EBSCO. This was followed by an assessment of maturity of the retrieved data. Principle-based evaluation was done to reveal epistemological, pragmatic, linguistic and logic principles, that attribute to the concept. Summative conclusions of each respective component and a detailed analysis of conceptual components (antecedents, attributes, outcomes, boundaries) resulted in a definition of woman-centered care. RESULTS: Eight studies were selected for analyses. In midwifery, woman-centered care has both a philosophical and a pragmatic meaning. There is strong emphasis on the woman-midwife relationship during the childbearing period. The concept demonstrates a dual and equal focus on physical parameters of pregnancy and birth, and on humanistic dimensions in an interpersonal context. The concept is epistemological, dynamic and multidimensional. The results reveal the concept's boundaries and fluctuations regarding equity and control. The role of the unborn child is not incorporated in the concept. CONCLUSION: An in-depth understanding and a broad conceptual foundation of womancentered care has evolved. Now, the concept is ready for research and educational purposes as well as for practical utility.

Viral dsRNA-activated human dendritic cells produce IL-27, which selectively promotes cytotoxicity in naive CD8+ T cells.

Viral recognition programs DCs to express Signal 3 molecules that promote the differentiation of effector CD8(+) T cells. Besides IL-12, another DC-derived IL-12 family member, IL-27, has been reported to contribute herein, but its specific role is not well understood. Here, we show that whereas IL-12 potently induces inflammatory cytokines (i.e., IFN-γ and TNF-α, but not IL-2), IL-27 excels in inducing proliferation and a cytotoxic profile (GrB, cytotoxicity of target cells) in human naïve CD8(+) T cells. Compared with bacterial cell-wall peptidoglycan, viral dsRNA-mimic poly (I:C) is superior in priming human BDCA1(+) peripheral blood DCs to produce IL-12 and IL-27, which promote inflammatory cytokines and a cytotoxic profile in differentiating CD8(+) T cells, respectively. These data support the concept that viral dsRNA-activated human DCs produce IL-27 to act as a specialized procytotoxic, antiviral cytokine in the development of effector CD8(+) T cells.