Chloroplast envelope K+/H+ antiporters are involved in cytosol pH regulation.

Variations in light intensity induce cytosol pH changes in photosynthetic tissues, providing a possible signal to adjust a variety of biochemical, physiological and developmental processes to the energy status of the cells. It was shown that these pH changes are partially due to the transport of protons in or out of the thylakoid lumen. However, the ion transporters in the chloroplast that transmit these pH changes to the cytosol are not known. KEA1 and KEA2 are K+/H+ antiporters in the chloroplast inner envelope that adjust stromal pH in light-to-dark transitions. We previously determined that stromal pH is higher in kea1kea2 mutant cells. In this study, we now show that KEA1 and KEA2 are required to attenuate cytosol pH variations upon sudden light intensity changes in leaf mesophyll cells, showing they are important components of the light-modulated pH signalling module. The kea1kea2 mutant mesophyll cells also have a considerably less negative membrane potential. Membrane potential is dependent on the activity of the plasma membrane proton ATPase and is regulated by secondary ion transporters, mainly potassium channels in the plasma membrane. We did not find significant differences in the activity of the plasma membrane proton pump but found a strongly increased membrane permeability to protons, especially potassium, of the double mutant plasma membranes. Our results indicate that chloroplast envelope K+/H+ antiporters not only affect chloroplast pH but also have a strong impact on cellular ion homeostasis and energization of the plasma membrane.

A self-unfolding proximity enabling device for oral delivery of macromolecules.

Oral delivery of macromolecules remains highly challenging due to their rapid degradation in the gastrointestinal tract and poor absorption across the tight junctions of the epithelium. In the last decade, researchers have investigated several medical devices to overcome these challenges using various approaches, some of which involve piercing through the intestine using micro and macro needles. We have developed a new generation of medical devices called self-unfolding proximity enabling devices, which makes it possible to orally deliver macromolecules without perforating the intestine. These devices protect macromolecules from the harsh conditions in the stomach and release their active pharmaceutical ingredients in the vicinity of the intestinal epithelium. One device version is a self-unfolding foil that we have used to deliver insulin and nisin to rats and pigs respectively. In our study, this device has shown a great potential for delivering peptides, with a significant increase in the absorption of solid dosage of insulin by ∼12 times and nisin by ∼4 times in rats and pigs, respectively. With the ability to load solid dosage forms, our devices can facilitate enhanced absorption of minimally invasive oral macromolecule formulations.

Recreational screen media use among Danish children aged 6-11 years: influence of parental screen media habits and attitudes.

AIMS: Little is known about the influence of parents' screen media habits and attitudes towards screen media on children's screen use. We investigated associations of parental screen use, their smartphone addiction and screen media attitudes, with children's recreational screen use. METHODS: This study was based on a population-based cross-sectional survey sent between May 2019 and November 2020 to a random sample of 6820 Danish parent-child dyads who answered questions regarding their screen media habits. Children were 6-11 years of age and had to reside with the parent. Multivariable adjusted regression analyses were conducted (in October 2021) separately for screen media use on weekdays and weekend days. RESULTS: The analyses included 5437 parents (41.0 years, 67.6% girls) and 5437 children (8.9 years, 48.2% girls). The adjusted relative odds of excessive amounts of screen use of children (>3 hours/weekday and >4 hours/weekend day) was 5.8 (95% confidence interval (CI) 4.6; 7.3) on weekdays and 7.2 (95% CI 5.9; 8.8) on weekend days comparing the fourth and first quartile of parental screen use. Children of parents in the fourth quartile of parental screen use had 2.1 (95% CI 1.7; 2.5) and 2.5 (95% CI 2.2; 3.0) greater odds of screen use before bedtime on all week and weekend days, respectively. Children of parents who had a positive attitude towards their child's screen use or were at high risk of smartphone addiction had significantly higher screen use and more frequent problematic screen use. CONCLUSIONS: Parent's screen media habits and attitudes were strongly associated with their children's recreational screen use.

Regulation of the mammalian-brain V-ATPase through ultraslow mode-switching.

Vacuolar-type adenosine triphosphatases (V-ATPases)1-3 are electrogenic rotary mechanoenzymes structurally related to F-type ATP synthases4,5. They hydrolyse ATP to establish electrochemical proton gradients for a plethora of cellular processes1,3. In neurons, the loading of all neurotransmitters into synaptic vesicles is energized by about one V-ATPase molecule per synaptic vesicle6,7. To shed light on this bona fide single-molecule biological process, we investigated electrogenic proton-pumping by single mammalian-brain V-ATPases in single synaptic vesicles. Here we show that V-ATPases do not pump continuously in time, as suggested by observing the rotation of bacterial homologues8 and assuming strict ATP-proton coupling. Instead, they stochastically switch between three ultralong-lived modes: proton-pumping, inactive and proton-leaky. Notably, direct observation of pumping revealed that physiologically relevant concentrations of ATP do not regulate the intrinsic pumping rate. ATP regulates V-ATPase activity through the switching probability of the proton-pumping mode. By contrast, electrochemical proton gradients regulate the pumping rate and the switching of the pumping and inactive modes. A direct consequence of mode-switching is all-or-none stochastic fluctuations in the electrochemical gradient of synaptic vesicles that would be expected to introduce stochasticity in proton-driven secondary active loading of neurotransmitters and may thus have important implications for neurotransmission. This work reveals and emphasizes the mechanistic and biological importance of ultraslow mode-switching.

Increased Soluble PD-1 Predicts Response to Nivolumab plus Ipilimumab in Melanoma.

BACKGROUND: Checkpoint inhibitors have revolutionized the treatment of metastatic melanoma, yielding long-term survival in a considerable proportion of the patients. Yet, 40-60% of patients do not achieve a long-term benefit from such therapy, emphasizing the urgent need to identify biomarkers that can predict response to immunotherapy and guide patients for the best possible treatment. Here, we exploited an unsupervised machine learning approach to identify potential inflammatory cytokine signatures from liquid biopsies, which could predict response to immunotherapy in melanoma. METHODS: We studied a cohort of 77 patients diagnosed with unresectable advanced-stage melanoma undergoing treatment with first-line nivolumab plus ipilimumab or pembrolizumab. Baseline and on-treatment plasma samples were tested for levels of PD-1, PD-L1, IFNγ, IFNß, CCL20, CXCL5, CXCL10, IL6, IL8, IL10, MCP1, and TNFα and analyzed by Uniform Manifold Approximation and Projection (UMAP) dimension reduction method and k-means clustering analysis. RESULTS: Interestingly, using UMAP analysis, we found that treatment-induced cytokine changes measured as a ratio between baseline and on-treatment samples correlated significantly to progression-free survival (PFS). For patients treated with nivolumab plus ipilimumab we identified a group of patients with superior PFS that were characterized by significantly higher baseline-to-on-treatment increments of PD-1, PD-L1, IFNγ, IL10, CXCL10, and TNFα compared to patients with worse PFS. Particularly, a high PD-1 increment was a strong individual predictor for superior PFS (HR = 0.13; 95% CI 0.034-0.49; p = 0.0026). In contrast, decreasing levels of IFNγ and IL6 and increasing levels of CXCL5 were associated with superior PFS in the pembrolizumab group, although none of the cytokines were individually predictors for PFS. CONCLUSIONS: In short, our study demonstrates that a high increment of PD-1 is associated with superior PFS in advanced-stage melanoma patients treated with nivolumab plus ipilimumab. In contrast, decreasing levels of IFNγ and IL6, and increasing levels of CXCL5 are associated with response to pembrolizumab. These results suggest that using serial samples to monitor changes in cytokine levels early during treatment is informative for treatment response.

Effects of Limiting Recreational Screen Media Use on Physical Activity and Sleep in Families With Children: A Cluster Randomized Clinical Trial.

Importance: Children and adults spend large amounts of their leisure time using screen media, which may affect their health and behavior. Objective: To investigate the effect of reducing household recreational screen media use on physical activity and sleep in children and adults. Design, Setting, and Participants: This was a cluster randomized clinical trial with a 2-week follow-up. Enrollment began on June 6, 2019, and ended on March 30, 2021. This study included a population-based sample from 10 Danish municipalities. A total of 89 families (181 children and 164 adults) were recruited based on a population-based survey on screen media habits in families with children. To be eligible, the responding parent had to list self-reported recreational screen use greater than the 40th percentile of recreational screen time use in the source population (>2.4 hours per day). In addition, the parent had to be full-time employed (with no regular night shifts) or enrolled in full-time education. Interventions: Families were randomly assigned to the screen media reduction intervention (45 families, 86 children, 82 adults) designed to ensure participant compliance to a maximum use of screen media (≤3 hours per week) for a 2-week period. Families randomly assigned to the control group (44 families, 95 children, 82 adults) were instructed to carry on as usual. Main Outcomes and Measures: The primary outcome was between-group difference in leisure nonsedentary activity (in minutes per day) measured by combined thigh and waist accelerometry. Secondary outcomes included other physical activity and sleep parameters measured by single-channel electroencephalography. Results: Among the 89 randomized families (intervention group [45 families]: 86 children; mean [SD] age, 8.6 [2.7] years; 44 boys [51%]; 42 girls [49%]; control group [44 families]: 95 children, mean [SD] age, 9.5 [2.5] years; 38 boys [40%]; 57 girls [60%]), 157 children (87%) had complete data on the primary outcome. Eighty-three children (97%) in the intervention group were compliant to the screen use reduction during the intervention. The mean (SD) change in leisure nonsedentary activity in the intervention group was 44.8 (63.5) minutes per day and in the control group was 1.0 (55.1) minute per day (intention-to-treat between-group mean difference, 45.8 minutes per day; 95% CI, 27.9-63.6 minutes per day; P < .001). No significant between-group mean differences were observed between intervention and control for the electroencephalography-based sleep outcomes. Conclusions and Relevance: In this cluster randomized clinical trial, a recreational screen media reduction intervention resulted in a substantial increase in children's engagement in physical activity. The large effect size suggests that the high levels of recreational screen media use seen in many children should be a public health concern. Trial Registration: ClinicalTrials.gov Identifier: NCT04098913.

TLR2 and TLR7 mediate distinct immunopathological and antiviral plasmacytoid dendritic cell responses to SARS-CoV-2 infection.

Understanding the molecular pathways driving the acute antiviral and inflammatory response to SARS-CoV-2 infection is critical for developing treatments for severe COVID-19. Here, we find decreasing number of circulating plasmacytoid dendritic cells (pDCs) in COVID-19 patients early after symptom onset, correlating with disease severity. pDC depletion is transient and coincides with decreased expression of antiviral type I IFNα and of systemic inflammatory cytokines CXCL10 and IL-6. Using an in vitro stem cell-based human pDC model, we further demonstrate that pDCs, while not supporting SARS-CoV-2 replication, directly sense the virus and in response produce multiple antiviral (interferons: IFNα and IFNλ1) and inflammatory (IL-6, IL-8, CXCL10) cytokines that protect epithelial cells from de novo SARS-CoV-2 infection. Via targeted deletion of virus-recognition innate immune pathways, we identify TLR7-MyD88 signaling as crucial for production of antiviral interferons (IFNs), whereas Toll-like receptor (TLR)2 is responsible for the inflammatory IL-6 response. We further show that SARS-CoV-2 engages the receptor neuropilin-1 on pDCs to selectively mitigate the antiviral interferon response, but not the IL-6 response, suggesting neuropilin-1 as potential therapeutic target for stimulation of TLR7-mediated antiviral protection.

Effects of limiting digital screen use on well-being, mood, and biomarkers of stress in adults.

Studies have linked higher digital screen use with poorer mental health. However, there is limited experimental evidence to suggest a causal relationship. In this trial, we aimed to investigate the effects of limiting recreational digital screen use on mental well-being, mood, and biomarkers of stress in healthy young and middle-aged adults. We randomly allocated 89 families (including 164 adults) to participate in an extensive screen media reduction intervention or control. Participants in the intervention group were instructed to decrease their recreational screen use to less than 3 hours/week/person. Intervention compliance was assessed using applications and tv-monitors. Overall subjective mental well-being and mood, and collected daily biomarkers of stress (salivary cortisol and cortisone) was assessed at baseline and 2-week follow-up. Reducing recreational digital screen use resulted in significantly improved self-reported well-being and mood in adults allocated to the intervention compared to control. We observed no intervention effects for biomarkers of stress. (ClinicalTrials.gov: NCT04098913, 23/09/2019).

Recreational screen media use in Danish school-aged children and the role of parental education, family structures, and household screen media rules.

Screen media use is part of most children's everyday lives, but organisations have advised that use should be limited. The aims of this study were to describe 6-11-year-old Danish children's screen device ownership and screen media use (weekdays and weekends), including the role of parental education, family structure and household screen media rules. We conducted a cross-sectional study including 5274 Danish children aged 6-11-years sampled from ten Danish municipalities from May 2019 to November 2020. Characteristics of the sample and source population were obtained from the Danish Health Data Authority. Parent's completed the SCREENS questionnaire, which was developed to assess children's screen media habits. We used inverse probability weighted logistic and linear regression models. Smartphone and laptop ownership was higher with increasing age, and use of screen media varied across day type, age and gender. The proportion of children using screen media more than 4 h/day was 13% (95% CI 12%;14%) for weekdays and 28% (95% CI 27%;29%) for weekend days. Children of parents with medium-length or long educations had statistically significant lower odds of using screen media more than 4 h/day. We found a statistically significant graded relationship between household screen media rules and children's screen media use; the less parents reported presence of rules, the more time their children spent on screen media engagements. Our results suggest that parental educational level and family structure are related to unfavourable screen media habits, and household screen media rules may play an important role for parents to limit children's screen use.

Cell-Extrinsic Priming Increases Permissiveness of CD4+ T Cells to Human Immunodeficiency Virus Infection by Increasing C-C Chemokine Receptor Type 5 Co-receptor Expression and Cellular Activation Status.

The chemokine receptor CCR5 is expressed on multiple cell types, including macrophages, dendritic cells, and T cells, and is the major co-receptor used during HIV transmission. Using a standard αCD3/CD28 in vitro stimulation protocol to render CD4+ T cells from PBMCs permissive to HIV infection, we discovered that the percentage of CCR5+ T cells was significantly elevated in CD4+ T cells when stimulated in the presence of peripheral blood mononuclear cells (PBMCs) as compared to when stimulated as purified CD4+ T cells. This indicated that environmental factors unique to the T-PBMCs condition affect surface expression of CCR5 on CD4+ T cells. Conditioned media from αCD3/CD28-stimulated PBMCs induced CCR5 expression in cultures of unstimulated cells. Cytokine profile analysis of these media suggests IL-12 as an inducer of CCR5 expression. Mass cytometric analysis showed that stimulated T-PBMCs exhibited a uniquely activated phenotype compared to T-Pure. In line with increased CCR5 expression and activation status in stimulated T-PBMCs, CD4+ T cells from these cultures were more susceptible to infection by CCR5-tropic HIV-1 as compared with T-Pure cells. These results suggest that in order to increase ex vivo infection rates of blood-derived CD4+ T cells, standard stimulation protocols used in HIV infection studies should implement T-PBMCs or purified CD4+ T cells should be supplemented with IL-12.

Manual Annotation of Time in Bed Using Free-Living Recordings of Accelerometry Data.

With the emergence of machine learning for the classification of sleep and other human behaviors from accelerometer data, the need for correctly annotated data is higher than ever. We present and evaluate a novel method for the manual annotation of in-bed periods in accelerometer data using the open-source software Audacity®, and we compare the method to the EEG-based sleep monitoring device Zmachine® Insight+ and self-reported sleep diaries. For evaluating the manual annotation method, we calculated the inter- and intra-rater agreement and agreement with Zmachine and sleep diaries using interclass correlation coefficients and Bland-Altman analysis. Our results showed excellent inter- and intra-rater agreement and excellent agreement with Zmachine and sleep diaries. The Bland-Altman limits of agreement were generally around ±30 min for the comparison between the manual annotation and the Zmachine timestamps for the in-bed period. Moreover, the mean bias was minuscule. We conclude that the manual annotation method presented is a viable option for annotating in-bed periods in accelerometer data, which will further qualify datasets without labeling or sleep records.

Feasibility of home-based sampling of salivary cortisol and cortisone in healthy adults.

OBJECTIVE: Salivary cortisol and cortisone are used as biomarkers of physiological stress. Careful sampling of saliva for profiling of awakening response and the diurnal slope can be challenging in free-living environments, and validated sampling protocols are lacking. Therefore, we investigated (1) the level of compliance to a three-day home-based salivary sampling protocol, and (2) the within subject day-to-day variability of cortisol and cortisone outcomes and the required measuring days to obtain high reproducibility. RESULTS: Nineteen healthy adults (mean age: 42, 50% females) participated. Participants collected in total 434 salivary samples out of 456 scheduled (four samples per day over three consecutive days at two time points). We found high level of compliance to the proposed free-living salivary sampling protocol with 18 (95%) and 16 (84%) participants being compliant to numbers and timing of samples, respectively. The area under the curve for the morning salivary samples and peak-to-bed slope had moderate reproducibility for cortisol and cortisone (intraclass correlation coefficient: 0.51-0.68, and mean coefficient of variation: 14.7%-75.3%). Three-to-four measuring days were required for high reproducibility of the area under the curve for the morning salivary samples and peak-to-bed slope using this free-living salivary sampling protocol. Trial registration Clinical trial registered with www.clinicaltrials.gov (NCT03788525).

Normal tissue complication probability models for prospectively scored late rectal and urinary morbidity after proton therapy of prostate cancer.

BACKGROUND AND PURPOSE: Photons and protons have fundamentally different properties, i.e. protons have a reduced dose bath but a higher relative biological effectiveness. Photon-based normal tissue complication probability (NTCP) models may therefore not immediately be applicable to proton therapy (PT). The aim was to derive parameters of the Lyman-Kutcher-Burman (LKB) NTCP model using prospectively recorded late morbidity data from PT, focusing on rectal morbidity and prostate cancer. MATERIALS AND METHODS: Prospectively collected data were available for 1151 prostate cancer patients treated with passive scattering PT and prescribed target doses of 78-82 Gy (RBE = 1.1) in 2 Gy fractions. Morbidity data (CTCAE v3.0) consisted of two alternative late grade 2 rectal bleeding endpoints: Medical Grade2A (GR2A) and procedural Grade2B (GR2B), as well as late grade 3 + urinary morbidity. GR2A + 2B were observed in 156/1047 patients (15%), GR2B in 45/1047 patients (4%), and urinary grade 3 + in 51/1151 patients (4%). LKB NTCP model parameters (D50, m, and n) were derived by maximum likelihood estimation. RESULTS: For the rectum/rectal wall the volume parameter n was low (0.07-0.14) for both GR2A + 2B and GR2B, as was the m parameter (range: 0.16-0.20). For the bladder/bladder wall both parameters were high (n-range: 0.20-0.36; m-range: 0.32-0.36). D50 parameters were higher for GR2B of the rectum/rectal wall (95.9-98.0 Gy) and bladder/bladder wall (118.1-119.9 Gy), but lower for GR2A2B (71.7-73.6 Gy). CONCLUSION: PT specific LKB NTCP model parameters were derived from a population of more than 1000 patients. The D50 parameter differed for all structures and endpoints and deviated from typical photon-based LKB model values.

Feasibility of two screen media reduction interventions: Results from the SCREENS pilot trial.

BACKGROUND: Advancements in screen media devices has transformed the way families engage with screen media. Although these modern devices offer many opportunities, e.g. communication and research online, an in-depth understanding of how these devices affect our health, is lacking. Before a definite randomized controlled trial, the SCREENS pilot study was conducted to assess compliance to and feasibility of two interventions, a measurement protocol, and a survey-based recruitment strategy. Also, the potential of the interventions to impact leisure time spent non-sedentary in children six-to-ten years of age was explored. METHODS: Families (N = 12) were recruited through a population-based survey sent out in October of 2018 to adults (N = 1,675) in the Municipality of Middelfart, Denmark. Families were randomized to one of two two-week interventions; an Evening Restriction intervention (no screen media use after six pm) and a General Restrict intervention (limit entertainment-based screen media to three hours/week/person). Intervention compliance was assessed objectively by measuring household TV usage, smartphone and tablet activity via an application, and via screen media diaries. During baseline and follow-up, as part of larger protocol, family members wore two triaxial accelerometers for seven consecutive days. The potential of the interventions to impact non-sedentary time was explored based on means and standard errors (SEs). RESULTS: Despite almost 85% and 75% reductions in leisure screen media use 0% and 50% of families were compliant in the Evening Restrict group and General Restrict group, respectively, based on strict a priori criteria. Participant feedback indicated that the General Restrict intervention generally was feasibly. Compliance to the accelerometry wear protocol was high (median non-wear was <1 hour/week). Moreover, the recruitment strategy was implemented and was feasible. The General restrict intervention might increase children's non-sedentary time (mean (SE): 36.6 (23) min/day, N = 6). CONCLUSIONS: The General Restriction intervention, the accelerometer wear protocol and recruitment strategy, appeared feasible. TRIAL REGISTRATION: NCT03788525 at https://clinicaltrials.gov [Retrospectively registered; 27th of December, 2018].

A Cognitive Functional Therapy+ Pathway Versus an Interdisciplinary Pain Management Pathway for Patients With Severe Chronic Low Back Pain (CONFeTTI Trial): Protocol for a Pragmatic Randomized Controlled Trial.

OBJECTIVE: Chronic low back pain (cLBP) is the leading cause of disability. Interdisciplinary pain management is recommended for patients with severe/high-impact cLBP. Such programs are expensive, not easily accessible, and have limited effect; therefore, new cost-effective strategies are warranted. Cognitive functional therapy (CFT) has shown promising results but has not been compared with an interdisciplinary pain management approach. The primary aim of this randomized controlled trial is to investigate if a pathway starting with CFT including psychologist support (CFT+) with the option of additional usual care (if needed) is superior in improving disability and more cost-effective at 12 months compared with an interdisciplinary pain management pathway (usual care). METHODS: This pragmatic, 2-arm, parallel-group randomized controlled trial will randomly allocate patients (n = 176) aged 18 to 75 years referred to an interdisciplinary pain center due to severe cLBP to 1 of 2 groups (1:1 ratio). Participants randomized to CFT+ will participate in a 3-month functional rehabilitation pathway with the option of additional usual care (if needed), and participants randomized to the interdisciplinary pain management pathway will participate in an individualized program of longer duration designed to best suit the individual's situation, needs, and resources. The primary outcome is the proportion of participants with an 8-point improvement in the Oswestry Disability Index score at 12 months. Exploratory outcomes are change in Oswestry Disability Index scores over time and an economic analysis of quality-adjusted life years using the 3-level version of the EuroQol EQ-5D. IMPACT: The study evaluates the cost-effectiveness of CFT+ with the option of additional usual care (if needed) for individuals with severe cLBP. Findings can potentially improve future care pathways and reduce cost for the health care system.

A conserved, buried cysteine near the P-site is accessible to cysteine modifications and increases ROS stability in the P-type plasma membrane H+-ATPase.

Sulfur-containing amino acid residues function in antioxidative responses, which can be induced by the reactive oxygen species generated by excessive copper and hydrogen peroxide. In all Na+/K+, Ca2+, and H+ pumping P-type ATPases, a cysteine residue is present two residues upstream of the essential aspartate residue, which is obligatorily phosphorylated in each catalytic cycle. Despite its conservation, the function of this cysteine residue was hitherto unknown. In this study, we analyzed the function of the corresponding cysteine residue (Cys-327) in the autoinhibited plasma membrane H+-ATPase isoform 2 (AHA2) from Arabidopsis thaliana by mutagenesis and heterologous expression in a yeast host. Enzyme kinetics of alanine, serine, and leucine substitutions were identical with those of the wild-type pump but the sensitivity of the mutant pumps was increased towards copper and hydrogen peroxide. Peptide identification and sequencing by mass spectrometry demonstrated that Cys-327 was prone to oxidation. These data suggest that Cys-327 functions as a protective residue in the plasma membrane H+-ATPase, and possibly in other P-type ATPases as well.

Screen-based media use and blood pressure in preschool-aged children: A prospective study in the Odense Child Cohort.

Aims: To examine prospective and cross-sectional associations between screen time and blood pressure (BP) in preschool children. Methods: The Odense Child Cohort study started in January 2010. Children who were born in the municipality of Odense underwent a clinical examination at 3 and 5 years of age and their parents were asked to complete a questionnaire. A total of 628 children were included in the prospective analysis and 964 children were included in two cross-sectional analyses at 5 years of age. Multivariable adjusted linear and logistic regression models were computed to examine prospective and cross-sectional associations between screen time and BP with adjustment for putative confounding factors. Results: No significant prospective association was found between a 2-year change in screen time and systolic BP (0.55 BP percentile change per 1 h increase in screen time, 95% confidence interval (CI) -1.51 to 2.60) and diastolic BP (0.74 BP percentile change per 1 h increase in screen time, 95% CI -1.09 to 2.57). No significant cross-sectional association was observed between screen time (⩽1 h/day, >1-2 h/day, >2 h/day) and the prevalence of high BP at 5 years of age. Exposure to screen time before bedtime 2-5 days/week and ⩾6 days/week was significantly associated with a greater prevalence of high BP compared with screen time before bedtime 0-1 day/week (odds ratios 1.57 (95% CI 1.02-2.42) and 1.82 (95% CI 1.18-2.89), respectively. Conclusions: No prospective association was found between screen time and BP. However, a significant cross-sectional association was found between screen time before bedtime and high BP in preschool children.

Mentalization-based treatment in groups for adolescents with Borderline Personality Disorder: 3- and 12-month follow-up of a randomized controlled trial.

Mentalization-based treatment in groups (MBT-G) has never been tested in adolescents with Borderline Personality Disorder (BPD) in a randomized controlled trial. The current study aimed to test the long-term effectiveness of MBT-G in an adolescent sample with BPD or BPD features (≥ 4 DSM-5 BPD criteria). Hundred and eleven patients with BPD (n = 106) or BPD features (n = 5) were randomized to either (1) a 1-year modified MBT-G program comprising three MBT introductory sessions, five individual case formulation sessions, 37 weekly MBT group sessions, and six MBT-Parent sessions, or (2) treatment as usual (TAU), defined as at least 12 individual monthly treatment sessions with follow-up assessments at 3 and 12 months post treatment. The primary outcome was the score on the Borderline Personality Features Scale for Children (BPFS-C), and secondary outcomes included clinician-rated BPD symptoms and global level of functioning as well as self-reported self-harm, depression, externalizing and internalizing symptoms, and caregiver reports. There were no statistically significant differences between MBT-G and TAU on the primary outcome measure or any of the secondary outcomes. Both groups showed improvement on the majority of clinical and social outcomes at both follow-up points, although remission rates were modest with just 35% in MBT-G and 39% in TAU 2 years after inclusion into the study. MBT-G was not superior to TAU in improving borderline features in adolescents. Although improvement was observed equally in both interventions over time, the patients continued to exhibit prominent BPD features, general psychopathology and decreased functioning in the follow-up period, which points to a need for more research and better understanding of effective components in early intervention programs. The ClinicalTrials.gov identifier is NCT02068326.

Predicted AS3MT Proteins Methylate Arsenic and Support Two Major Phylogenetic AS3MT Groups.

Inorganic arsenic is one of the most toxic and carcinogenic substances in the environment, but many organisms, including humans, methylate inorganic arsenic to mono-, di-, and trimethylated arsenic metabolites, which the organism can excrete. In humans and other eukaryotic organisms, the arsenite methyltransferase (AS3MT) protein methylates arsenite. AS3MT sequences from eukaryotic organisms group phylogenetically with predicted eubacterial AS3MT sequences, which has led to the suggestion that AS3MT was acquired from eubacteria by multiple events of horizontal gene transfer. In this study, we evaluated whether 55 (out of which 47 were predicted based on protein sequence similarity) sequences encoding putative AS3MT orthologues in 47 species from different kingdoms can indeed methylate arsenic. Fifty-three of the proteins showed arsenic methylating capacity. For example, the predicted AS3MT of the human gut bacterium Faecalibacterium prausnitzii methylated arsenic efficiently. We performed a kinetic analysis of 14 AS3MT proteins representing two phylogenetically distinct clades (Group 1 and 2) that each contain both eubacterial and eukaryotic sequences. We found that animal and bacterial AS3MTs in Group 1 rarely produce trimethylated arsenic, whereas Hydra vulgaris and the bacterium Rhodopseudomonas palustris in Group 2 produce trimethylated arsenic metabolites. These findings suggest that animals during evolution have acquired different arsenic methylating phenotypes from different bacteria. Further, it shows that humans carry two bacterial systems for arsenic methylation: one bacterium-derived AS3MT from Group 1 incorporated in the human genome and one from Group 2 in F. prausnitzii present in the gut microbiome.

Multivariate normal tissue complication probability models for rectal and bladder morbidity in prostate cancer patients treated with proton therapy.

BACKGROUND AND PURPOSE: Normal tissue complication probability (NTCP) models applied for model-based patient selection to proton therapy (PT) have usually been derived using dose/volume histogram (DVH) parameters from photon-based radiotherapy. This study aimed to derive PT-specific multivariate NTCP models that also accounted for the spatial dose distribution (rectum only) as well as non-dose/volume related factors. MATERIALS AND METHODS: The study included rectum and bladder DVHs, 2D rectal dose maps and relevant patient/treatment characteristics from 1151 prostate cancer cases treated with PT. Prospectively scored Grade 2 late rectal bleeding (CTCAE v3.0, also procedural interventions separately) (n = 156 (15%)) and Grade 3+ GU morbidity (n = 51 (4%)) were entered into a multivariate logistic regression analysis. Model evaluation included assessment of the area under the receiver operating characteristic curve (AUC). RESULTS: Anticoagulant use was a dominant predictor, chosen in four of the six rectum models and in the bladder model. Age was a dominant predictor in all procedural only rectum models while prostate volume, bladder D5% and V75Gy were predictors in the bladder model. The selection frequency of the dose/volume predictors varied widely, where the percentage of the anterior rectum surface receiving >=75 Gy was the most robust. AUC values ranged from 0.58 to 0.70 across all models, with no clear difference between the DVH- and spatial-based models for the rectum. CONCLUSION: Anticoagulant use and age were the most prominent predictors in the NTCP models. V75Gy of the rectal wall and the bladder was a predictor in the DVH-based models of the rectum and bladder respectively.