Deep Sequencing and Phenotyping in an Australian Tuberous Sclerosis Complex "No Mutations Identified" Cohort.

Tuberous sclerosis complex (TSC) is a variable multisystem disorder. The "no mutations identified" (NMI) group are reportedly phenotypically milder than those with an identified molecular cause, and often have mosaic or intronic variants not detected by standard sequencing methods. METHODS: We describe the phenotypes in an Australian TSC NMI group (n = 18) and a molecular testing strategy implementable in a diagnostic laboratory. Massively parallel sequencing (MPS) of the whole genomic regions of TSC1 and TSC2 was performed using DNA extracted from multiple tissue samples per participant. RESULTS: Our study showed that the phenotype in TSC NMI individuals can be similar to those with heterozygous, particularly TSC1, variants. Although neurodevelopmental outcomes can be less severe, the number of organ systems involved was similar to the non-mosaic groups. A diagnostic yield of 72% (13/18) was achieved, with the majority (10/13) being mosaic variants and the remainder heterozygous variants missed on previous testing. CONCLUSION: Testing DNA from multiple tissue samples allowed for validation of otherwise discarded low-level mosaic variants and detection of mosaic variants by MPS without excessive cost or the need for specialised techniques. Implementing this approach in a diagnostic setting is viable and allows optimal clinical care of patients with NMI TSC.

Biaryl Anion Radical Formation by Potassium Metal Reduction of Aryl Isocyanates and Triaryl Isocyanurates.

The potassium metal reduction of aryl isocyanates (aryl = phenyl, p-tolyl, 3,5-dimethylphenyl, 4-biphenylyl, and 1-naphthyl) in THF with 18-crown-6 or in HMPA results in the formation of the corresponding triaryl isocyanurate anion radicals. Continued exposure to potassium results in loss of the isocyanurate anion radical and the eventual formation of the respective biaryl anion radical. The 1,1'-binaphthyl anion radical is found to undergo a cyclodehydrogenation reaction, which leads to formation of the perylene anion radical. When authentic triaryl isocyanurates are reduced with metal, the heterocyclic ring undergoes fragmentation with elimination of carbon monoxide to produce a triarylbiuret dianion. This ring opening reaction is initiated by the two-electron reduction of the neutral isocyanurate species. The biaryl anion radical is formed when the biuret dianion is reduced further with metal. A possible mechanism for biaryl formation involves a heterolytic cleavage of an aryl C-N bond and release of an aryl radical once the triarylbiuret dianion is further reduced. A subsequent intermolecular reaction between two aryl radicals forms the corresponding biaryl, which can then be reduced to the anion radical. Notably, when a mixture of two different triaryl isocyanurate compounds is reduced in solution, the corresponding mixed biaryl anion radical is generated.

Oral health status and factors associated with oral health of primary school children in Gulu district, northern Uganda.

BACKGROUND: Globally, oral diseases remain a major public health problem. However, there is limited information about the oral health status and factors associated with oral disease among children in Uganda. The aim of this study was to examine the oral health status and factors associated with oral health of primary school children in urban and rural areas of the Gulu district of northern Uganda. METHODS: A comparative cross-sectional study was conducted among 356 school children aged 11-13 years attending six schools located in urban and rural areas. The children received a clinical oral examination and participated in a questionnaire survey that collected information on sociodemographic and oral health knowledge, attitude, and practices. All data were entered and analysed using IBM SPSS Statistics for Windows, Version 26.0. Armonk, NY: IBM Corp statistical software. Logistic regression analyses examined factors associated with dental caries and gingival bleeding. RESULTS: Of the 356 children (11-13 years) included, the mean age was 12.2 years, 140 (39.3%) were male and 176 (49.4%) were from urban areas. The proportion of school children with dental caries was 33.6% (n = 119), with the mean decayed, missing due to caries, and filled teeth (DMFT) index of 0.81 (25th percentile = 0; 75th percentile = 1.00). There was no significant difference in caries prevalence between rural and urban children (31.6% versus 35.6%, p = 0.33). Of the children involved in the study, 141(39.8%) had gum bleeding. The mean oral knowledge score was 2.85 ± 1.53 (range, 0-7), while the mean attitude, hygiene practice, frequency of sweets consumption, and oral health related impact scores were 4.25 ± 1.23 (range, 1-6), 5.40 ± 1.81 (range, 0-9), 25.66 ± 4.29 (range 9-54) and 2.1 ± 1.65 (range, 0-6), respectively. Using logistic regression analyses, as oral health knowledge score increased the odds of not having dental caries increased (aOR = 1.19, 95% CI 1.02-1.39). CONCLUSION: The prevalence of dental caries and gum bleeding of primary school children in Gulu district is high. Children lacked knowledge on causes of oral disease, and behaviour towards oral disease prevention. In addition, oral health knowledge scores were significantly associated with dental caries. Oral health education programs in schools should emphasise providing skills-based education.

Effect of perioperative management on early graft function in living donor paediatric kidney transplantation.

BACKGROUND: Paediatric kidney transplantation has an increased risk of surgical and vascular complications, with intensive care monitoring required postoperatively. This study aimed to determine if perioperative management affects early graft function in living donor paediatric kidney transplantation. METHODS: Clinical data was extracted from the electronic medical record for living donor kidney transplants at two paediatric centres covering the state of New South Wales (NSW), Australia from 2009 to 2021. Estimated glomerular filtration rate (eGFR) of 7 days and 1-month post-transplant were calculated as measures of early graft function. RESULTS: Thirty-nine eligible patients (female n (%) 13 (33%)) with a median (IQR) age of 6 (3-9) years and pre-transplant eGFR of 7 (6-10) mL/min/1.73 m2 were analysed. Mean (SD) central venous pressure (CVP) after revascularisation was 11 (4) mmHg. Intraoperatively, mean volume of fluid administered was 84 (39) mL/kg, and 34 (87%) patients received vasoactive agents. Average systolic blood pressure (BP) in the first 24-h post-transplant was 117 (12) mmHg. Postoperatively, median volume of fluid administered in the first 24 h was 224 (159-313) mL/kg, and 17 (44%) patients received vasoactive agents. Median eGFR 7 days and 1-month post-transplant were 115 (79-148) and 103 (83-115) mL/min/1.73 m2, respectively. Linear regression analyses demonstrated that after adjusting for age, the average CVP after revascularisation and average systolic BP in the first 24-h post-transplant were not associated with eGFR in the first month post-transplant. CONCLUSIONS: Targeted intraoperative and postoperative fluid and haemodynamic characteristics were achieved but did not correlate with early graft function.

The Role of Humeral Torsion on Glenohumeral Rotation in Injured Baseball Players.

Background: Differences in rotational range of motion (ROM) compared to humeral retrotorsion (HRT)-corrected rotational ROM exist in healthy baseball athletes, but it is unclear whether these differences exist in a pathological population. Purpose/Hypothesis: The purpose of this study was to determine if there are disparities between objectively measured differences in ROM and HRT-corrected deficits in injured baseball players. It was hypothesized that disparities would exist between (1) the side-to-side difference in glenohumeral external rotation (GER) and the HRT-corrected glenohumeral external rotation deficit (GERD) and (2) the side-to-side difference in glenohumeral internal rotation (GIR) and the HRT-corrected glenohumeral internal rotation deficit (GIRD). Study Design: Cross-sectional study; Level of evidence, 3. Methods: Data from 172 baseball players with shoulder or elbow injuries (45 shoulder, 127 elbow) were reviewed in July 2023. GER and GIR were measured on the injured and noninjured sides of all players, and diagnostic ultrasound was used to measure HRT. Dependent t tests were run to compare the side-to side differences in GER and GIR with the HRT-corrected GERD and GIRD, respectively. Results: In the players with a shoulder injury, there was a significant disparity between the side-to-side difference in GER and the HRT-corrected GERD (2°± 14° vs -13°± 15°, respectively) and between the side-to-side difference in GIR and the HRT-corrected GIRD (-14°± 8° vs 2°± 9°, respectively) (P < .001 for both). Similarly, players with an elbow injury had significant disparities between the side-to-side difference in GER and the HRT-corrected GERD (6°± 9° vs -10°± 9°, respectively) and between the side-to-side difference in GIR and the HRT-corrected GIRD (-12°± 8° vs 4°± 10°, respectively) (P < .001 for both). Conclusion: The results supported our hypothesis that there were disparities between objectively measured differences in GER and GIR compared with the HRT-corrected GERD and GIRD in injured baseball players. Consideration must be given to osseous adaptations that occur at the glenohumeral joint when evaluating and treating this population.

The randomized controlled trial (NAVKIDS2) of a patient navigator program created for children with chronic kidney disease.

Patient navigators enable adult patients to circumnavigate complex health systems, improving access to health care and outcomes. Here, we aimed to evaluate the effects of a patient navigation program in children with chronic kidney disease (CKD). In this multi-center, randomized controlled trial, we randomly assigned children (aged 0-16 years) with CKD stages 1-5 (including children on dialysis or with kidney transplants), from low socioeconomic status backgrounds, and/or residing in remote areas, to receive patient navigation at randomization (immediate) or at six months (waitlist). The primary outcome was self-rated health (SRH) of participating children at six months, using intention to treat analysis. Secondary outcomes included caregivers' SRH and satisfaction with health care, children's quality of life, hospitalizations, and missed school days. Repeated measures of the primary outcome from baseline to six months were analyzed using cumulative logit mixed effects models. Semi-structured interviews were thematically evaluated. Of 398 screened children, 162 were randomized (80 immediate and 82 waitlist); mean age (standard deviation) of 8.8 (4.8) years with 64.8% male. SRH was not significantly different between the immediate and wait-listed groups at six months. There were also no differences across all secondary outcomes between the two groups. Caregivers' perspectives were reflected in seven themes: easing mental strain, facilitating care coordination, strengthening capacity to provide care, reinforcing care collaborations, alleviating family tensions, inability to build rapport and unnecessary support. Thus, in children with CKD, self-rated health may not improve in response to a navigator program, but caregivers gained skills related to providing and accessing care.

A narrative review of little league shoulder: proximal humeral physis widening is only one piece of the puzzle, it is time to consider posterior glenoid dysplasia.

Baseball athletes across all levels of play are at an increased risk for upper extremity injury due to the supraphysiologic demands on the shoulder and elbow during overhead throwing. Little league baseball players present with a unique subset of injuries that can affect the growth plate, commonly at the shoulder or the elbow. Ascertaining a diagnosis and plan of care for little league shoulder (LLS) historically focuses on the proximal humeral physis in skeletally immature throwing athletes presenting with shoulder pain. However, while not a current standard of care, posterior glenoid dysplasia is often present in youth baseball athletes presenting with LLS, warranting a shift in the way clinicians evaluate for and treat the youth baseball athlete's pathologic shoulder. Therefore, purpose of this narrative review is 2-fold: first, to describe the current standard of care as it relates to a diagnosis of LLS, and second, to critically describe a comprehensive evaluation process for youth throwing athletes with shoulder pain that includes screening for evidence of posterior glenoid dysplasia. This paper summarizes the current state of the available evidence for anatomic considerations of LLS in the baseball athletes throwing shoulder. Additionally, we provide a framework for clinical evaluation using a multidisciplinary approach to evaluate the entire kinetic chain of the youth baseball athlete presenting with LLS and posterior glenoid dysplasia. A case study is presented to describe common presentations, clinical and objective examinations, and a plan of care from time of evaluation to return to throwing.

MANOCCA: a robust and computationally efficient test of covariance in high-dimension multivariate omics data.

Multivariate analysis is becoming central in studies investigating high-throughput molecular data, yet, some important features of these data are seldom explored. Here, we present MANOCCA (Multivariate Analysis of Conditional CovAriance), a powerful method to test for the effect of a predictor on the covariance matrix of a multivariate outcome. The proposed test is by construction orthogonal to tests based on the mean and variance and is able to capture effects that are missed by both approaches. We first compare the performances of MANOCCA with existing correlation-based methods and show that MANOCCA is the only test correctly calibrated in simulation mimicking omics data. We then investigate the impact of reducing the dimensionality of the data using principal component analysis when the sample size is smaller than the number of pairwise covariance terms analysed. We show that, in many realistic scenarios, the maximum power can be achieved with a limited number of components. Finally, we apply MANOCCA to 1000 healthy individuals from the Milieu Interieur cohort, to assess the effect of health, lifestyle and genetic factors on the covariance of two sets of phenotypes, blood biomarkers and flow cytometry-based immune phenotypes. Our analyses identify significant associations between multiple factors and the covariance of both omics data.

Medical student interactions with children in their daily lives.

Learning to communicate effectively with children in clinical interactions can be challenging. This study aimed to determine the extent to which medical students are exposed to children in their daily lives, in order to understand the experience students bring when entering paediatric rotations. METHODS: A cross-sectional survey of medical students entering paediatric rotations from two medical schools was conducted. Students were asked to rate the frequency of their interactions with infants, preschool-aged and school-aged children and their confidence in doing so. RESULTS: 339 out of 476 students participated in this study. Interactions with infants and preschool-aged children were rare, with most students reporting interactions once or two times per year or less (83% and 67%, respectively). Students interacted with school-aged children more frequently (43% most weeks or days). Students who interacted more frequently with children were more confident when entering their paediatric placements. CONCLUSIONS: Medical students have limited exposure to infants and preschool-aged children in their daily lives and this affects their confidence. Supervisors should incorporate activities aimed at building confidence interacting with young children early in clinical attachments.

Psychological wellbeing among parents of a child living with a serious chronic illness: A cross-sectional survey study.

Parents of a child with a chronic illness can experience greater distress than the average population, yet little is understood about differences between illness groups. This cross-sectional survey study aimed to compare parents' psychological distress and perceived wellbeing across five chronic illnesses. Parents from one Australian pediatric hospital completed the Kessler Psychological Distress Scale and seven purpose-designed items about their wellbeing. Data from 106 parents (cancer = 48, cystic fibrosis [CF] = 27, kidney disease = 12, gastrointestinal condition/disorder = 9, developmental and epileptic encephalopathy [DEE] = 10) was analysed using bivariate Pearson's Correlation and linear mixed-effects models. Parents' distress scores differed between groups (F(4,80) = 2.50, p = .049), with the DEE group reporting higher distress than the CF group (mean difference = 6.76, 95% CI [0.11, 13.42]). Distress scores were moderately correlated to parents' perceptions of their child's health and their own wellbeing. Parents' self-reported coping with their child's condition/treatments differed (F(4,81) = 3.24, p = .016), with the DEE group rating their coping as poorer than the CF group (mean difference = -25.32, 95% CI [-46.52, 4.11]). Across all groups, parents reported unmet needs, particularly for psychosocial support and practical/financial assistance. Support interventions may be most effective if tailored to the child's illness, with greater support potentially needed for parents who have a child with DEE and/or severe comorbidities.

Are serious games seriously good at preparing students for clinical practice?: A randomized controlled trial.

PURPOSE: Serious games (SGs) have great potential for pediatric medical education. This study evaluated the efficacy of a SG in improving learner satisfaction, knowledge, and behavior. MATERIALS AND METHODS: This was an investigator-blinded randomized controlled trial (RCT) comparing a SG against two controls: (i) adaptive tutorial (AT), and (ii) low-stimulus control (LSC). SG is a highly immersive role-playing game in a virtual hospital. AT delivers interactive web-based lessons. LSC is paper-based clinical practice guidelines. Metropolitan senior medical students at UNSW were eligible. A total of 154 enrolled and were block randomized to one intervention. Participants had access to one intervention for 8 weeks which taught pediatric acute asthma and seizure assessment and management. Satisfaction was assessed with Likert-scale responses to 5 statements and 2 free-text comments. Knowledge was assessed with 10 multiple-choice questions (MCQs). Clinical behavior was assessed during a 30-point simulated clinical management scenario (CMS). Primary analysis was performed on a modified intention-to-treat basis and compared: (1) SG vs. AT; and (2) SG vs. LSC. RESULTS: A total of 118 participants were included in the primary analysis (modified intention-to-treat model). No significant differences in MCQ results between the SG and control groups. SG group outperformed the LSC group in the CMS, with a moderate effect (score out of 30: 20.8 (3.2) vs. 18.7 (3.2), respectively, d = 0.65 (0.2-1.1), p = 0.005). No statistically significant difference between SG and AT groups in the CMS (score: 20.8 (3.2) vs. 19.8 (3.1), respectively, d = 0.31 (-0.1 to 0.8), p = 0.18). A sensitivity analysis (per-protocol model) was performed with similar outcomes. CONCLUSIONS: This is the first investigator-blinded RCT assessing the efficacy of a highly immersive SG on learner attitudes, knowledge acquisition, and performance in simulated pediatric clinical scenarios. The SG demonstrated improved translation of knowledge to a simulated clinical environment, particularly compared to LSC. SGs show promise in pediatric medical education.

Upper Extremity Musculoskeletal Profiles in Tennis Players: A Systematic Review.

CONTEXT: Tennis-specific musculoskeletal (MSK) screening can assess range of motion (ROM) and muscular imbalances. Identifying normative values before implementing a MSK screen is essential in contributing to athlete performance and injury risk profiles. OBJECTIVE: To review upper extremity MSK data in healthy tennis players across age, sex, and level of play. DATA SOURCE: The Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines were followed for this review. A search was conducted in MEDLINE, SPORTDiscus, Embase, and CINAHL. STUDY SELECTION: This review included shoulder, elbow, and wrist ROM, isometric strength, or isokinetic strength in a tennis population. Each article was critically appraised to help identify the internal and external validity of each study. STUDY DESIGN: Systematic review. LEVEL OF EVIDENCE: Level 3. DATA EXTRACTION: A total of 41 studies met the search criteria. Each contributor organized the data elements of interest into data tables, with a second contributor assigned for review. Data elements of interest included player and study characteristics: ROM, isometric dynamometry, and isokinetic strength. RESULTS: A total of 3174 players were included in the final studies. Most of the players included were competitive adolescents and young adults; 15 studies included ROM data. Male tennis players consistently had more external rotation (ER) gain (range, 1.8º to 8.8º) and internal rotation (IR) loss (range, -15.3º to -3.0º) when compared with their female counterparts (ER range, -2.5º to 5.8º; IR range, -10.4º to -3º). Shoulder IR and ER strength were measured in the majority of all the strength studies, with the external rotators generating at least two-thirds the strength of the internal rotators. CONCLUSION: Overall MSK data of tennis players indicate that shoulder strength values are often larger than nontennis players, but equal to or slightly lower than comparable athletes in other overhead sports. Adaptive changes of the glenohumeral joint and subsequent rotational motion are similar to those of other overhead athletes.

The colibactin-producing Escherichia coli alters the tumor microenvironment to immunosuppressive lipid overload facilitating colorectal cancer progression and chemoresistance.

Intratumoral bacteria flexibly contribute to cellular and molecular tumor heterogeneity for supporting cancer recurrence through poorly understood mechanisms. Using spatial metabolomic profiling technologies and 16SrRNA sequencing, we herein report that right-sided colorectal tumors are predominantly populated with Colibactin-producing Escherichia coli (CoPEC) that are locally establishing a high-glycerophospholipid microenvironment with lowered immunogenicity. It coincided with a reduced infiltration of CD8+ T lymphocytes that produce the cytotoxic cytokines IFN-γ where invading bacteria have been geolocated. Mechanistically, the accumulation of lipid droplets in infected cancer cells relied on the production of colibactin as a measure to limit genotoxic stress to some extent. Such heightened phosphatidylcholine remodeling by the enzyme of the Land's cycle supplied CoPEC-infected cancer cells with sufficient energy for sustaining cell survival in response to chemotherapies. This accords with the lowered overall survival of colorectal patients at stage III-IV who were colonized by CoPEC when compared to patients at stage I-II. Accordingly, the sensitivity of CoPEC-infected cancer cells to chemotherapies was restored upon treatment with an acyl-CoA synthetase inhibitor. By contrast, such metabolic dysregulation leading to chemoresistance was not observed in human colon cancer cells that were infected with the mutant strain that did not produce colibactin (11G5∆ClbQ). This work revealed that CoPEC locally supports an energy trade-off lipid overload within tumors for lowering tumor immunogenicity. This may pave the way for improving chemoresistance and subsequently outcome of CRC patients who are colonized by CoPEC.

Targeted Metagenomic Databases Provide Improved Analysis of Microbiota Samples.

We report on Moonbase, an innovative pipeline that builds upon the established tools of MetaPhlAn and Kraken2, enhancing their capabilities for more precise taxonomic detection and quantification in diverse microbial communities. Moonbase enhances the performance of Kraken2 mapping by providing an efficient method for constructing project-specific databases. Moonbase was evaluated using synthetic metagenomic samples and compared against MetaPhlAn3 and generalized Kraken2 databases. Moonbase significantly improved species precision and quantification, outperforming marker genes and generalized databases. Construction of a phylogenetic tree from 16S genome data in Moonbase allowed for the incorporation of UniFrac-type phylogenetic information into diversity calculations of samples. We demonstrated that the resulting analysis increased statistical power in distinguishing microbial communities. This study highlights the continual evolution of metagenomic tools with the goal of improving metagenomic analysis and highlighting the potential of the Moonbase pipeline.

The 2022 world health organization reevaluation of human and mammalian toxic equivalency factors for polychlorinated dioxins, dibenzofurans and biphenyls.

In October 2022, the World Health Organization (WHO) convened an expert panel in Lisbon, Portugal in which the 2005 WHO TEFs for chlorinated dioxin-like compounds were reevaluated. In contrast to earlier panels that employed expert judgement and consensus-based assignment of TEF values, the present effort employed an update to the 2006 REP database, a consensus-based weighting scheme, a Bayesian dose response modeling and meta-analysis to derive "Best-Estimate" TEFs. The updated database contains almost double the number of datasets from the earlier version and includes metadata that informs the weighting scheme. The Bayesian analysis of this dataset results in an unbiased quantitative assessment of the congener-specific potencies with uncertainty estimates. The "Best-Estimate" TEF derived from the model was used to assign 2022 WHO-TEFs for almost all congeners and these values were not rounded to half-logs as was done previously. The exception was for the mono-ortho PCBs, for which the panel agreed to retain their 2005 WHO-TEFs due to limited and heterogenous data available for these compounds. Applying these new TEFs to a limited set of dioxin-like chemical concentrations measured in human milk and seafood indicates that the total toxic equivalents will tend to be lower than when using the 2005 TEFs.

Sleep disturbances in children and adolescents after kidney transplantation.

BACKGROUND: Disturbances of sleep are prevalent among children with chronic kidney disease. However, the aetiology of sleep disorders in children particularly after kidney transplantation is not clear. We sought to ascertain the prevalence and type of sleep disturbances in paediatric kidney transplant recipients and to identify predictors of sleep disturbances in this population. METHODS: Caregivers of kidney transplant recipients completed online questionnaires about their child's sleep. The questionnaires utilised were the Sleep Disturbance Scale for Children (SDSC), the Epworth Sleepiness Scale for Children and Adolescents (ESS-CHAD), questions about sleep hygiene, and questions about restless leg syndrome. Demographic and clinical details were collected from medical records. RESULTS: Thirty-five children were included in the study, with a median (IQR) age of 14.1 years (9.5-16.1) and median years (IQR) since transplant of 3.7 (0.7-8.7) years, and 72.0% were identified to have at least one category of sleep disturbance according to scores on the SDSC. The most common sleep disturbances reported were disorders of initiating and maintaining sleep (DIMS) (40.0%) and disorders of excessive somnolence (DOES) (31.4%). Statistically significant predictors of sleep disturbances include low estimated glomerular filtration rate and increased age. Among children who screened positive for DIMS and DOES, the majority indicated use of electronic devices in 1 h before bed. CONCLUSIONS: A high prevalence of sleep disturbances has been identified in children after kidney transplants, and some risk factors may be modifiable. Further studies are required to understand whether there are other readily modifiable predictors of sleep disturbances.

Does the Nutritional Intake and Diet Quality of Children With Chronic Kidney Disease Differ From Healthy Controls? A Comprehensive Evaluation.

OBJECTIVE: Children with chronic kidney disease (CKD) experience many obstacles to achieving optimal dietary intake. Dietary intake patterns remain unexplored or poorly described. This study compares nutritional intake and diet quality of Australian children with CKD to controls. METHODS: A food frequency questionnaire captured intake data and was compared to controls. Nutritional intake was determined using individualized nutrient reference values, and diet quality described using the Australian Guide to Healthy Eating and the Australian Child and Adolescent Recommended Food Score. RESULTS: Children with CKD (n = 36) and controls (n = 82) were studied. Children with CKD had lower weight and height z scores, but higher body mass index (P < .0001 for all parameters). Children with CKD had adequate energy intake, and excessive protein and sodium intake (336% and 569%). They were significantly less likely to meet requirements for vitamin A (P < .001), thiamine (P = .006), folate (P = .01), vitamin C (P = .008), calcium (P < .0001), iron (P = .01), magnesium (P = .0009), and potassium (P = .002). No child met recommended vegetable intake; however, less than half of children with CKD met fruit (44%), grains (31%), and dairy serves (31%). They were also less likely to meet recommended fruit and dairy serves (P = .04 and P = .01, respectively). Non-core foods provided 36% of energy, and although comparable to controls, was contributed more by takeaway foods (P = .01). CONCLUSION: Children with CKD have reduced nutritional intake of key nutrients and consume more takeaways than controls. Attention to increasing core foods, limiting sodium intake, and managing restrictions while promoting nutrient density appears necessary.

Perceptions of oral health promotion in primary schools among health and education officials, community leaders, policy makers, teachers, and parents in Gulu district, northern Uganda: A qualitative study.

INTRODUCTION: One in every two cases of caries in deciduous teeth occurs in low- and middle-income countries (LMICs). The aim of the World Health Organisation's (WHO) Healthy Schools Program is to improve the oral health of children. This study explored perceptions of implementation of the Ugandan oral health schools' program in Gulu district, northern Uganda. METHODS: Semi-structured interviews were conducted with a purposive sample of 19 participants including health and education officials, community leaders, policy makers, teachers, and parents. All interviews were transcribed verbatim and analysed thematically. RESULTS: Our study identified three themes: (1) components of oral health promotion, (2) implementation challenges of oral health promotion, and (3) development of an oral health policy. The components of oral health promotion in schools included engagement of health workers, the community, companies, skills-based education, and oral health services. Participants were concerned about insufficient funding, unsatisfactory skills-based education, and inadequate dental screening. Participants reported that there was an urgent need to develop oral health policy to guide implementation of the program at scale. CONCLUSIONS: Schools provided oral health promotion that aligned with existing features of the WHO's health-promoting school framework. Implementation of this strategy could be enhanced with increased resources, adequate oral health education, and explicit development of oral health policy.

Hospital and urban wastewaters shape the matrix and active resistome of environmental biofilms.

Understanding the dynamics of antibiotic resistance gene (ARG) transfer and dissemination in natural environments remains challenging. Biofilms play a crucial role in bacterial survival and antimicrobial resistance (AMR) dissemination in natural environments, particularly in aquatic systems. This study focused on hospital and urban wastewater (WW) biofilms to investigate the potential for ARG dissemination through mobile genetic elements (MGEs). The analysis included assessing the biofilm extracellular polymeric substances (EPS), microbiota composition as well as metatranscriptomic profiling of the resistome and mobilome. We produced both in vitro and in situ biofilms and performed phenotypic and genomic analyses. In the in vitro setup, untreated urban and hospital WW was used to establish biofilm reactors, with ciprofloxacin added as a selective agent at minimal selective concentration. In the in situ setup, biofilms were developed directly in hospital and urban WW pipes. We first showed that a) the composition of EPS differed depending on the growth environment (in situ and in vitro) and the sampling origin (hospital vs urban WW) and that b) ciprofloxacin impacted the composition of the EPS. The metatranscriptomic approach showed that a) expression of several ARGs and MGEs increased upon adding ciprofloxacin for biofilms from hospital WW only and b) that the abundance and type of plasmids that carried individual or multiple ARGs varied depending on the WW origins of the biofilms. When the same plasmids were present in both, urban and hospital WW biofilms, they carried different ARGs.  We showed that hospital and urban wastewaters shaped the structure and active resistome of environmental biofilms, and we confirmed that hospital WW is an important hot spot for the dissemination and selection of antimicrobial resistance. Our study provides a comprehensive assessment of WW biofilms as crucial hotspots for ARG transfer. Hospital WW biofilms exhibited distinct characteristics, including higher eDNA abundance and expression levels of ARGs and MGEs, highlighting their role in antimicrobial resistance dissemination. These findings emphasize the importance of understanding the structural, ecological, functional, and genetic organization of biofilms in anthropized environments and their contribution to antibiotic resistance dynamics.

Optimizing Bioink Composition for Human Chondrocyte Expression of Lubricin.

The surface zone of articular cartilage is the first area impacted by cartilage defects, commonly resulting in osteoarthritis. Chondrocytes in the surface zone of articular cartilage synthesize and secrete lubricin, a proteoglycan that functions as a lubricant protecting the deeper layers from shear stress. Notably, 3D bioprinting is a tissue engineering technique that uses cells encapsulated in biomaterials to fabricate 3D constructs. Gelatin methacrylate (GelMA) is a frequently used biomaterial for 3D bioprinting cartilage. Oxidized methacrylated alginate (OMA) is a chemically modified alginate designed for its tunable degradation rate and mechanical properties. To determine an optimal combination of GelMA and OMA for lubricin expression, we used our novel high-throughput human articular chondrocyte reporter system. Primary human chondrocytes were transduced with PRG4 (lubricin) promoter-driven Gaussia luciferase, allowing for temporal assessment of lubricin expression. A lubricin expression-driven Design of Experiment screen and subsequent validation identified 14% GelMA/2% OMA for further study. Therefore, DoE optimized 14% GelMA/2% OMA, 14% GelMA control, and 16% GelMA (total solid content control) were 3D bioprinted. The combination of lubricin protein expression and shape retention over the 22 days in culture, successfully determined the 14% GelMA/2%OMA to be the optimal formulation for lubricin secretion. This strategy allows for rapid analysis of the role(s) of biomaterial composition, stiffness or other cell manipulations on lubricin expression by chondrocytes, which may improve therapeutic strategies for cartilage regeneration.