Copper Micro-Flowers for Electrocatalytic Sensing of Nitrate Ions in Water.

The progressive increase in nitrate's (NO3-) presence in surface and groundwater enhances environmental and human health risks. The aim of this work is the fabrication and characterization of sensitive, real-time, low-cost, and portable amperometric sensors for low NO3- concentration detection in water. Copper (Cu) micro-flowers were electrodeposited on top of carbon screen-printed electrodes (SPCEs) via cyclic voltammetry (with voltage ranging from -1.0 V to 0.0 V at a scan rate of 0.1 V s-1). The obtained sensors exhibited a high catalytic activity toward the electro-reduction in NO3-, with a sensitivity of 44.71 µA/mM. They had a limit of detection of 0.87 µM and a good dynamic linear concentration range from 0.05 to 3 mM. The results were compared to spectrophotometric analysis. In addition, the devices exhibited good stability and a maximum standard deviation (RSD) of 5% after ten measurements; reproducibility, with a maximum RSD of 4%; and repeatability after 10 measurements with the RSD at only 5.63%.

African Swine Fever Virus Protein-Protein Interaction Prediction.

The African swine fever virus (ASFV) is an often deadly disease in swine and poses a threat to swine livestock and swine producers. With its complex genome containing more than 150 coding regions, developing effective vaccines for this virus remains a challenge due to a lack of basic knowledge about viral protein function and protein-protein interactions between viral proteins and between viral and host proteins. In this work, we identified ASFV-ASFV protein-protein interactions (PPIs) using artificial intelligence-powered protein structure prediction tools. We benchmarked our PPI identification workflow on the Vaccinia virus, a widely studied nucleocytoplasmic large DNA virus, and found that it could identify gold-standard PPIs that have been validated in vitro in a genome-wide computational screening. We applied this workflow to more than 18,000 pairwise combinations of ASFV proteins and were able to identify seventeen novel PPIs, many of which have corroborating experimental or bioinformatic evidence for their protein-protein interactions, further validating their relevance. Two protein-protein interactions, I267L and I8L, I267L__I8L, and B175L and DP79L, B175L__DP79L, are novel PPIs involving viral proteins known to modulate host immune response.

CRISPR screens identify genes essential for in vivo virulence among proteins of hyperLOPIT-unassigned subcellular localization in Toxoplasma.

The research field to identify and characterize genes essential for in vivo virulence in Toxoplasma gondii has been dramatically advanced by a series of in vivo clustered regularly interspaced short palindromic repeats (CRISPR) screens. Although subcellular localizations of thousands of proteins were predicted by the spatial proteomic method called hyperLOPIT, those of more than 1,000 proteins remained unassigned, and their essentiality in virulence was also unknown. In this study, we generated two small-scale gRNA libraries targeting approximately 600 hyperLOPIT-unassigned proteins and performed in vivo CRISPR screens. As a result, we identified several genes essential for in vivo virulence that were previously unreported. We further characterized two candidates, TgGTPase and TgRimM, which are localized in the cytoplasm and the apicoplast, respectively. Both genes are essential for parasite virulence and widely conserved in the phylum Apicomplexa. Collectively, our current study provides a resource for estimating the in vivo essentiality of Toxoplasma proteins with previously unknown localizations.IMPORTANCEToxoplasma gondii is a protozoan parasite that causes severe infection in immunocompromised patients or newborns. Toxoplasma possesses more than 8,000 genes; however, the genes essential for in vivo virulence were not fully identified. The apicomplexan parasites, including Toxoplasma, developed unique organelles that do not exist in other model organisms; thus, determining the subcellular location of parasite proteins is important for understanding their functions. Here, we used in vivo genetic screens that enabled us to investigate hundreds of genes in Toxoplasma during mouse infection. We screened approximately 600 parasite proteins with previously unknown subcellular localizations. We identified many novel genes that confer parasite virulence in mice. Among the top hits, we characterized two genes essential for in vivo virulence, TgGTPase and TgRimM, which are widely conserved in the phylum Apicomplexa. Our findings will contribute to understanding how apicomplexans adapt to the host environment and cause disease.

Small molecule FICD inhibitors suppress endogenous and pathologic FICD-mediated protein AMPylation.

The AMP transferase, FICD, is an emerging drug target finetuning stress signaling in the endoplasmic reticulum (ER). FICD is a bi-functional enzyme, catalyzing both AMP addition (AMPylation) and removal (deAMPylation) from the ER resident chaperone BiP/GRP78. Despite increasing evidence linking excessive BiP/GRP78 AMPylation to human diseases, small molecules to inhibit pathogenic FICD variants are lacking. Using an in-vitro high-throughput screen, we identify two small-molecule FICD inhibitors, C22 and C73. Both molecules significantly inhibit FICD-mediated BiP/GRP78 AMPylation in intact cells while only weakly inhibiting BiP/GRP78 deAMPylation. C22 and C73 also efficiently inhibit pathogenic FICD variants and improve proinsulin processing in ß cells. Our study identifies and validates FICD inhibitors, highlighting a novel therapeutic avenue against pathologic protein AMPylation.

A genome-wide CRISPR/Cas9 screen identifies calreticulin as a selective repressor of ATF6α.

Activating transcription factor 6 (ATF6) is one of three endoplasmic reticulum (ER) transmembrane stress sensors that mediate the unfolded protein response (UPR). Despite its crucial role in long-term ER stress adaptation, regulation of ATF6 alpha (α) signalling remains poorly understood, possibly because its activation involves ER-to-Golgi and nuclear trafficking. Here, we generated an ATF6α/Inositol-requiring kinase 1 (IRE1) dual UPR reporter CHO-K1 cell line and performed an unbiased genome-wide CRISPR/Cas9 mutagenesis screen to systematically profile genetic factors that specifically contribute to ATF6α signalling in the presence and absence of ER stress. The screen identified both anticipated and new candidate genes that regulate ATF6α activation. Among these, calreticulin (CRT), a key ER luminal chaperone, selectively repressed ATF6α signalling: Cells lacking CRT constitutively activated a BiP::sfGFP ATF6α-dependent reporter, had higher BiP levels and an increased rate of trafficking and processing of ATF6α. Purified CRT interacted with the luminal domain of ATF6α in vitro and the two proteins co-immunoprecipitated from cell lysates. CRT depletion exposed a negative feedback loop implicating ATF6α in repressing IRE1 activity basally and overexpression of CRT reversed this repression. Our findings indicate that CRT, beyond its known role as a chaperone, also serves as an ER repressor of ATF6α to selectively regulate one arm of the UPR.

Measurement Method Options to Investigate Digital Screen Technology Use by Children and Adolescents: A Narrative Review.

The role and potential impact of digital screen technology in the lives of children is heavily debated. Current evidence is limited by the weakness of measures typically used to characterise screen use, predominantly proxy- or self-reports with known inaccuracy and bias. However, robust and detailed evidence is needed to provide practical trustworthy guidance to families and professionals working with families. The purpose of this paper is to support researchers to select measurement method(s) that will provide robust and detailed evidence. The paper outlines the challenges in measuring contemporary screen use by children, using a child-technology interaction model to organise considerations. A range of different methods used to measure digital screen technology use in children and adolescents (i.e., questionnaires, diaries, electronically prompted sampling, direct observation, fixed room cameras, wearable/portable cameras, audio recorders, screen-device onboard logging, remote digital trace logging and proximity logging) are described along with examples of their use and constructs typically measured as well as a summary of the advantages and disadvantages of each method. A checklist and worked examples are provided to support researchers determining the best methods or combination of methods for a research project.

Impact of Digital Media, School Problems, and Lifestyle Factors on Youth Psychosomatic Health: A Cross-Sectional Survey.

BACKGROUND: Post-pandemic psychosomatic complaints in children and adolescents have been underreported. This study investigated psychosomatic complaints in children and adolescents in Northern Italy in 2023, with the aim of identifying changes in predictors and vulnerable subgroups. METHODS: Cross-sectional data representative of scholars from a northern Italian province were analyzed using the Health Behavior in School-aged Children (HBSC) checklist. The sum scores, count data, and dichotomized data were examined, and predictor effects were expressed using a linear regression model for the sum scores. RESULTS: Data from 4525 participants (aged 7-19 years, 50.5% girls) were analyzed. Psychosomatic symptoms remained unchanged after the pandemic. Significant gender differences were noted, especially in older age groups, with girls reporting more complaints. Factors such as children's health, digital media use, and school problems significantly influence psychosomatic outcomes. While migration background negatively affected girls' psychosomatic well-being, boys showed improvement through sports. CONCLUSION: The psychosomatic well-being of children and adolescents did not improve after the pandemic. These findings indicate the need for targeted interventions, improved health literacy, and ongoing monitoring to support the mental well-being of this vulnerable population. Reducing screen time remains a critical strategy for enhancing youth well-being.

Examination of the Life Habits of Preschool Children Based on Their Screen Use Patterns.

Children who open their eyes to a digital world begin interacting with screens in the early years of life. The interaction between screens and children starts from the very first moments of life and intensifies over time. The aim of this study is to examine the effects of screen use patterns on the life habits of preschool children. In this context, the relationships between problematic media use, eating habits, and self-care skills among preschool children are analyzed from various perspectives. Structured according to a mixed-methods research approach, this study employs both qualitative and quantitative research designs. The study group consists of 582 children continuing their education in preschool institutions across seven different regions and nine different provinces in Turkey, along with 20 of their parents. Random and non-random sampling methods were used to form the study group. Quantitative data were collected using the Problematic Media Use Measure, Children's Eating Behavior Inventory, and Preschool Children (36-72 months) Self-Care Skills Scale-Teacher Form, while qualitative data were gathered through a semi-structured parent interview form. The study results indicate that problematic media use and eating behaviors significantly vary according to the screen time of children, with an increase in problematic media use linked to a rise in negative eating behaviors. Parents are found to perceive the use of media devices during mealtime as a necessity, thus employing them, and believe that their children's social behaviors are shaped according to screen content.

Leisure Time Habits and Levels of Physical Activity in Children and Adolescents.

BACKGROUND/OBJECTIVES: Childhood and adolescence are important stages of life for acquiring healthy habits. There is a high prevalence of sedentary lifestyles worldwide during these ages, which negatively impacts health. This is attributed, in part, to excessive time spent engaging in sedentary behaviors. The aim of this study was to assess the time spent on sedentary behaviors and their relationship with physical activity levels in children and adolescents in the Community of Madrid. METHODS: A total of 26,729 participants aged 10-17 from various schools and institutes took part in this study. The International Physical Activity Questionnaire Short Form (IPAQ-SF) was used, and they were asked about the time they spent on different leisure time habits (specific sedentary behaviors and organized physical activity). A generalized linear model was used to analyze the association between the time spent in sedentary activities and the time spent in physical activity. RESULTS: The results revealed that children and adolescents engage in low levels of physical activity and most of them spend considerable time in sedentary behaviors such as studying, watching television, or using social media. Completing school homework (Coef: 1.23, 95% CI: -0.51 to 2.97, p = 0.167) or using social media for more than 2 h (Coef: 1.29, 95% CI: -2.98 to 0.40, p = 0.133) compared to not dedicating time to them did not show a significant association with daily physical activity time. Watching television for more than 2 h was associated with a decrease of 2.60 min (95% CI: -4.41 to -0.78, p = 0.005). Thus, no or only irrelevant associations were found between time spent in sedentary activities and physical activity time. CONCLUSIONS: Despite the drawbacks of spending time engaging in sedentary behaviors, they seem to be compatible with physical activity levels. Therefore, it is important to continue research on physical activity adherence strategies to promote overall health and well-being.

The Correlation between Motor Skill Proficiency and Academic Performance in High School Students.

The intricate relationship between physical health and cognitive development has been a focal point of multidisciplinary research, suggesting that motor skill proficiency could influence academic outcomes. This study aimed to investigate the correlation between motor control, mobility, stability-measured by the Functional Movement Screen (FMS)-and academic performance among high school students. Conducted with 201 participants from a public high school in Spain, this descriptive non-experimental research divided students into high and low academic performance groups based on their average grade scores, with the median used as the criterion for distinction. The FMS was utilized to assess fundamental motor skills, and academic performance was verified across mathematics, language, science, and physical education subjects. The findings revealed a significant positive relationship between the FMS scores and academic achievement (r = 0.691; p < 0.001), indicating that students with higher motor skill proficiency had higher academic achievement. This correlation persisted across the various subjects, highlighting the potential benefits of integrating physical education and motor skill development into educational strategies. The study's results suggest that educational policies should advocate for comprehensive physical education programs to foster both physical well-being and academic improvement, thereby promoting a holistic educational model that enhances student performance across disciplines.

Estimation of Individual Positive Anti-Islet Autoantibodies from 3 Screen ICA Titer.

The 3 Screen ICA ELISA is a novel assay capable of simultaneously measuring autoantibodies to glutamic acid decarboxylase (GADA), insulinoma-associated antigen-2 (IA-2A), and zinc transporter 8 (ZnT8A), making it a valuable tool for screening type 1 diabetes. Despite its advantages, it cannot specify which individual autoantibodies are positive or negative. This study aimed to estimate individual positive autoantibodies based on the 3 Screen ICA titer. Six hundred seventeen patients with type 1 diabetes, simultaneously measured for 3 Screen ICA and three individual autoantibodies, were divided into five groups based on their 3 Screen ICA titer. The sensitivities and contribution rates of the individual autoantibodies were then examined. The study had a cross-sectional design. Sixty-nine percent (424 of 617) of patients with type 1 diabetes had 3 Screen ICA titers exceeding the 99th percentile cut-off level (20 index). The prevalence of GADA ranged from 80% to 100% in patients with a 3 Screen ICA over 30 index and 97% of patients with a 3 Screen ICA ≥300 index. Furthermore, the prevalence of all individual autoantibodies being positive was 0% for ≤80 index and as high as 92% for ≥300 index. Significant associations were observed in specific titer groups: the 20-29.9 index group when all the individual autoantibodies were negative, the 30-79.9 index group when positive for GADA alone or IA-2A alone, the 30-299.9 index group when positive for ZnT8A alone, the 80-299.9 index group when positive for both IA-2A and ZnT8A, the 300-499.9 index group when positive for both GADA and ZnT8A, and the ≥300 index group when positive for all individual autoantibodies. These results suggest that the 3 Screen ICA titer may be helpful in estimating individual positive autoantibodies.

Rapid iPSC inclusionopathy models shed light on formation, consequence, and molecular subtype of α-synuclein inclusions.

The heterogeneity of protein-rich inclusions and its significance in neurodegeneration is poorly understood. Standard patient-derived iPSC models develop inclusions neither reproducibly nor in a reasonable time frame. Here, we developed screenable iPSC "inclusionopathy" models utilizing piggyBac or targeted transgenes to rapidly induce CNS cells that express aggregation-prone proteins at brain-like levels. Inclusions and their effects on cell survival were trackable at single-inclusion resolution. Exemplar cortical neuron α-synuclein inclusionopathy models were engineered through transgenic expression of α-synuclein mutant forms or exogenous seeding with fibrils. We identified multiple inclusion classes, including neuroprotective p62-positive inclusions versus dynamic and neurotoxic lipid-rich inclusions, both identified in patient brains. Fusion events between these inclusion subtypes altered neuronal survival. Proteome-scale α-synuclein genetic- and physical-interaction screens pinpointed candidate RNA-processing and actin-cytoskeleton-modulator proteins like RhoA whose sequestration into inclusions could enhance toxicity. These tractable CNS models should prove useful in functional genomic analysis and drug development for proteinopathies.

Adherence of Brazilian Minors to the 24-Hour Movement Guidelines after In-Person School Return.

The levels of adequacy of movement behaviors after returning to in-person school activities following the COVID-19 pandemic are not yet well understood. This study aimed to assess the adherence of Brazilian minors to the recommendations of the 24-Hour Movement Guidelines (moderate to vigorous physical activity, recreational screen time, and sleep duration), as well as overall adherence to these guidelines, after the relaxation of COVID-19 social isolation measures and the resumption of in-person schooling. A cross-sectional study was conducted with parents or guardians (39 ± 7.8) of minors aged up to 18 years of age (3.8 ± 2.5). A total of 172 individuals responded to the questionnaire. Data were compared with those obtained in the Survey of the Adequacy of Brazilian Children and Adolescents to the 24-Hour Movement Guidelines before and during the COVID-19 Pandemic. There was an 18.6 percentage decrease (p-value < 0.001) in overall adherence to the 24-Hour Movement Guidelines when comparing the periods before the COVID-19 pandemic (March 2020) and after the return to in-person schooling (March 2021). The largest percentage drop in adherence between these periods was observed for moderate to vigorous physical activity (27.4%; p-value < 0.001) and the lowest for sleep (10.5%; p-value < 0.001). Adherence to the 24-Hour Movement Guidelines did not return to pre-pandemic levels with the resumption of in-person school activities.

Parental Mental Health, Gender, and Lifestyle Effects on Post-Pandemic Child and Adolescent Psychosocial Problems: A Cross-Sectional Survey in Northern Italy.

BACKGROUND: The exacerbation of psychosocial problems among children and adolescents during the coronavirus disease 2019 (COVID-19) pandemic necessitates an assessment of the long-term mental health impact of pandemic interventions. Focusing on both pandemic-related factors and demographic variables, such as gender and daily habits, an analysis was conducted to understand how these elements continue to affect young populations in the post-pandemic era. METHODS: In April 2023, a comprehensive online survey was administered to families in South Tyrol, Italy, with children aged 7-19 years, to ensure age and gender representation. The survey included parent ratings and adolescent (11-19 years) self-reports using standardized instruments to measure the symptoms of mental health problems (Strengths and Difficulties Questionnaire, SDQ), anxiety (Screen for Child Anxiety Related Emotional Disorders, SCARED), and depression (Patient Health Questionnaire-2, PHQ-2). Statistical analyses included descriptive statistics, chi-square tests, and unadjusted odds ratios (ORs). RESULTS: Of the 4525 valid responses, 1831 were self-reported by adolescents. Notable gender differences in mental health outcomes were identified, along with significant demographic predictors, such as age, single parenthood, parental mental health problems, and immigrant background. Negative effects were associated with reduced family climate and increased screen time, whereas physical activity showed beneficial effects. Proxy reports overestimated adolescents' mental health problems, whereas self-reports tended to underestimate them. CONCLUSIONS: Persistent mental health problems and gender disparities highlight the need for a public health approach. This should include accessible support services, resilience building, targeted support for vulnerable families and gender-specific interventions.

Association of Screen Time Exposure With Autism Spectrum Disorder in Four to Six-Year-Old Children in Arar City, Saudi Arabia.

Background Autism spectrum disorder (ASD) is a psychopathologic disorder caused by several factors. The early signs include poor interaction and communication, delayed milestones, and repeated behavior patterns. This study aimed to assess the relationship between screen time and ASD severity and investigate the types of electronic devices associated with ASD in children aged four to six years in Arar City, Kingdom of Saudi Arabia (KSA). Methodology A cross-sectional study was conducted in primary healthcare centers (PHCs) in Arar City, KSA. The study enrolled all parents with children aged four to six years attending the PHCs in Arar City, KSA. Results The total sample size was 199 participants. Regarding the relationship between screen time exposure and ASD, there were variable screen time exposure durations, with 22.6% of children exposed for less than an hour, 30.7% for one to two hours, and 46.7% for more than two hours. Moreover, the type of electronic devices to which children were exposed varied, with smartphones being the most prevalent (68.3%). In terms of the age of children since exposure to electronic devices, the data indicated that 30.2% were exposed before the age of two, 35.2% between two and three years, and 34.7% after three years of age. Regarding the relationship with sociodemographic characteristics, there was a statistically significant relationship with the mother's age at birth (p = 0.050), mother's education level (p = 0.009), father's education level (p = 0.049), whether the child was suffering from any chronic or neurological disease (p = 0.008), age since the child was exposed to electronic devices (p = 0.049), and screen time exposure duration (p = 0.040). Conclusions The study highlights the significant association between screen time exposure and the development of ASD in children. Public awareness of this associated risk among caregivers is recommended to follow the protective guidelines. Further research and interventions are needed to better understand and address the impact of screen media use on children's neurodevelopment and overall well-being.

Organic Cation Modulation in Manganese Halides to Optimize Crystallization Process and X-Ray Response Toward Large-Area Scintillator Screen.

Manganese halides are one of the most potential candidates for large-area flat-panel detection owing to their biological safety and all-solution preparation. However, reducing photon scattering and enhancing the efficient luminescence of scintillator screens remains a challenge due to their uncontrollable crystallization and serious nonradiative recombination. Herein, an organic cation modulation is reported to control the crystallization process and enhance the luminescence properties of manganese halides. Given the industrial requirements of the X-ray flat-panel detector, the large-area A2MnBr4 screen (900 cm2) with excellent uniformity is blade-coated at 60 °C. Theoretical calculations and in situ measurements reveal that organic cations with larger steric hindrance can slow down the crystallization of the screen, thus neatening the crystal arrangement and reducing the photon scattering. Moreover, larger steric hindrance can also endow the material with higher exciton binding energy, which is beneficial for restraining nonradiative recombination. Therefore, the BPP2MnBr4 (BPP = C25H22P+) screen with larger steric hindrance exhibits a superior spatial resolution (>20 lp mm-1) and ultra-low detection limit (< 250 nGyair s-1). This is the first time steric hindrance modulation is used in blade-coated scintillator screens, and it believes this study will provide some guidance for the development of high-performance manganese halide scintillators.

Associations of 24-hour movement behaviors with emotional and behavioral problems among Chinese adolescents.

PURPOSE: The role of adherence to the recommendations for 24-hour movement behaviors (24-HMB), including physical activity (PA), screen time (ST), and sleep duration (SLP), in relation to emotional and behavioral problems in Chinese adolescents remains uncertain. This study aimed to investigate these associations and explore potential sex differences. METHODS: This school-based cross-sectional study included 15,071 Chinese adolescents with a mean age of 14.53 (SD: 1.65) years. Data on emotional and behavioral problems and 24-HMB (including PA, ST, and SLP) were collected. Analysis was performed using general linear mixed models, with additional sex-stratified analyses conducted. RESULTS: The number of 24-HMB recommendations met was negatively associated with total difficulties (ß estimate=-0.96, 95% CI: -1.07 to -0.85) and positively related to prosocial behavior (ß estimate = 0.41, 95% CI: 0.37 to 0.46) among adolescents. Compared with none of the recommendations met, meeting all recommendations (total difficulties: ß estimate=-2.98, 95% CI: -3.41 to -2.55; prosocial behaviors: ß estimate = 1.05, 95%CI: 0.87 to 1.24) demonstrated the strongest association with both difficulties and prosocial behaviors, followed by meeting recommendations for PA + ST (total difficulties: ß estimate=-2.15, 95% CI: -2.41 to -1.90; prosocial behaviors: ß estimate = 0.98, 95% CI: 0.87 to 1.09). These associations were consistently significant in both boys and girls. CONCLUSION: Adherence to more 24-HMB recommendations, particularly meeting all recommendations or combined PA + ST recommendations, could improve emotional and behavioral well-being among adolescent girls and boys. The significance of balanced movement behaviors for promoting adolescent mental health merits increased attention.

Screen Use Time and Its Association With Mental Health Issues in Young Adults in India: Protocol for a Cross-Sectional Study.

BACKGROUND: Screen use time has increased in the past decade owing to the increased availability and accessibility of digital devices and the internet. Several studies have shown an association between increased screen use time and mental health issues such as anxiety and depression. However, studies in the young adult population-a demographic with high screen use-and in low- and middle-income country settings are limited. OBJECTIVE: This protocol describes a study that aims to measure self-reported screen use times and patterns in young adults (18-24 y) in India and assess if increased screen use time is associated with poorer mental well-being. METHODS: This protocol describes a cross-sectional study of a pan-India, web-based convenience sample of young adults (18-24 y) with access to digital devices with a screen and a minimum of secondary school education. Participants will be recruited through people in the professional networks of the investigators, which includes pediatricians. The survey will also be distributed via the social media pages of our organization (X [X Corp], Instagram [Meta], Facebook [Meta], etc). Sociodemographic details will be collected through a questionnaire designed by the authors; screen use time and patterns will be assessed using an adaptation of the Screen Time Questionnaire to include data on different apps and websites used on digital devices; and mental health parameters will be gauged using the Warwick-Edinburgh Mental Well-Being Scale, Generalized Anxiety Disorder Scale, Perceived Stress Scale, and Patient Health Questionnaire. For statistical analysis, we will consider the following variables: (1) the primary independent variable is screen use time; (2) other independent variables include age, gender, residence: rural or urban, educational qualifications, employment status, stress associated with familial financial status, average sleep time, number of people living in a house or rooms in that house, BMI, substance use, and past psychiatric history; and (3) dependent variables include mental well-being, depression, anxiety, and perceived stress. To quantify the association between screen use time and mental health, we will perform a Bayesian multivariate multiple regression analysis that models the possibility of multiple alternative hypotheses while accounting for relevant sociodemographic covariables. RESULTS: The survey instrument has been designed, and feedback has been obtained from the domain experts and members of our organization whose profile is similar to the potential study participants. The final data received after this study has been conducted will be analyzed and shared. As of January 2023, we have not yet initiated the data collection. CONCLUSIONS: Based on the findings of this study, we will be able to establish a correlation between device- and use-specific screen use time and various mental health parameters. This will provide a direction to develop screen use time and mental health guidelines among young adults. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): PRR1-10.2196/39707.

A random mutagenesis screen enriched for missense mutations in bacterial effector proteins.

To remodel their hosts and escape immune defenses, many pathogens rely on large arsenals of proteins (effectors) that are delivered to the host cell using dedicated translocation machinery. Effectors hold significant insight into the biology of both the pathogens that encode them and the host pathways that they manipulate. One of the most powerful systems biology tools for studying effectors is the model organism, Saccharomyces cerevisiae. For many pathogens, the heterologous expression of effectors in yeast is growth inhibitory at a frequency much higher than housekeeping genes, an observation ascribed to targeting conserved eukaryotic proteins. Abrogation of yeast growth inhibition has been used to identify bacterial suppressors of effector activity, host targets, and functional residues and domains within effector proteins. We present here a yeast-based method for enriching for informative, in-frame, missense mutations in a pool of random effector mutants. We benchmark this approach against three effectors from Legionella pneumophila, an intracellular bacterial pathogen that injects a staggering >330 effectors into the host cell. For each protein, we show how in silico protein modeling (AlphaFold2) and missense-directed mutagenesis can be combined to reveal important structural features within effectors. We identify known active site residues within the metalloprotease RavK, the putative active site in SdbB, and previously unidentified functional motifs within the C-terminal domain of SdbA. We show that this domain has structural similarity with glycosyltransferases and exhibits in vitro activity consistent with this predicted function.

LeishGEM: genome-wide deletion mutant fitness and protein localisations in Leishmania.

LeishGEM is a genome-wide functional annotation community resource for Leishmania mexicana, where deletion mutant growth in vitro and in vivo is measured and protein localisation is determined by endogenous tagging and LOPIT-DC (localisation of organelle proteins by isotope tagging with differential centrifugation) spatial proteomics. Data are being made available pre-publication via http://leishgem.org which allows data-driven identification of the mechanisms for Leishmania parasitism.