In patients with proximal hypospadias, often no genetic cause is identified despite extensive genetic testing. Many genes involved in sex development encode transcription factors with strict timing and dosing of the gene products. We hypothesised that there might be recurrent differences in DNA methylation in boys with hypospadias and that these might differ between patients born small versus appropriate for gestational age. Genome-wide Methylated DNA sequencing (MeD-seq) was performed on 32bp LpnPI restriction enzyme fragments after RE-digestion in leucocytes from 16 XY boys with unexplained proximal hypospadias, one with an unexplained XX testicular disorder/difference of sex development (DSD) and twelve, healthy, sex- and age-matched controls. Five of seven differentially methylated regions (DMRs) between patients and XY controls were in the Long Intergenic Non-Protein Coding RNA 665 (LINC00665; CpG24525). Three patients showed hypermethylation of MAP3K1. Finally, no DMRs in XX testicular DSD associated genes were identified in the XX boy versus XX controls. In conclusion, we observed no recognizable epigenetic signature in 16 boys with XY proximal hypospadias and no difference between children born small versus appropriate for gestational age. Comparison to previous methylation studies in individuals with hypospadias did not show consistent findings, possibly due to the use of different inclusion criteria, tissues and methods.

Medical education is experiencing a paradigm shift towards more interactive and collaborative pedagogical approaches. Barcamps, also known as unconferences, offer an interactive, participant-driven learning approach. This study aims to evaluate the feasibility of using barcamps as an educational model in medical education. Furthermore, the possibility of establishing barcamps in medical education as a pedagogical tool is discussed. The study integrates two evaluations to achieve a multifaceted understanding: a pilot study conducted in 2022 and a subsequent feasibility study in 2023. Participation in the barcamps and their evaluations was voluntary. We combined quantitative data, primarily from post-event evaluations, and qualitative data from open-ended survey questions. These methods were designed to capture a broad range of participant experiences and perceptions. The pilot study in 2022 included 11 participants and produced a response rate for the evaluations. The feasibility study in 2023 had 34 participants but a lower evaluation response rate of 53%. Both barcamps were generally positively evaluated by the participants, indicating a high level of satisfaction and perceived value. Regarding the active participation of participants, the wide range of presented topics highlights the adaptability and flexibility of the barcamp model. Attendees at the pilot mentioned a significantly higher previous experience with barcamps. The study suggests that barcamps are generally well received as an interactive and collaborative educational tool in medical education, reflected by high participant evaluation scores. The wide range of topics presented and discussed indicates that barcamps can accommodate diverse educational needs and interests. However, the study also identified areas for improvement, most notably in the structure of topic plans and the overall learning environment. Finally, the principals for barcamps might adapt to other educational methods by adding more interaction, choice, collaboration, communication, critical thinking, creativity, and caring to the learning process.

UNASSIGNED: The aim was to evaluate the effectiveness of a marine omega fatty acid-containing multimodal wound matrix (MWM) in reducing bacterial contamination and supporting wound area reduction (WAR) in patients with hard-to-heal wounds of varying aetiologies.
UNASSIGNED: A prospective, single-site, pilot case series of patients with hard-to-heal wounds. All wounds were considered non-healing prior to inclusion as they had failed to achieve at least 50% WAR after at least four weeks of standard of care (SoC) treatments. Patients were seen once weekly for wound assessments, matrix application and dressing changes. Baseline and weekly fluorescence images, standard wound images and wound measurements were obtained.
UNASSIGNED: A total of three patients, two with venous leg ulcers (VLUs) and one with a diabetic foot ulcer (DFU) were enrolled in this pilot study. The mean baseline wound age prior to study enrolment was 24 weeks, with a mean baseline wound size of 8.61cm2. The two VLUs went on to complete closure. The DFU displayed a total WAR of 53% by six weeks, when the patient was lost to follow-up due to a geographical relocation. The mean percentage area reduction of all wounds combined was 82% upon study completion.
UNASSIGNED: The use of MWM proved to be effective and safe in this patient cohort. The wounds included in this case series failed to enter a healing trajectory with SoC wound therapies. The MWM supported wound closure and reduced bacterial loads in this patient cohort.

The pathogenesis of COVID-19 warrants unravelling. Genetic polymorphism analysis may help answer the variability in disease outcome. To determine the role of KIR and HLA polymorphisms in susceptibility, progression, and severity of SARS-CoV-2 infection, 458 patients and 667 controls enrolled in this retrospective observational study from April to December 2020. Mild/moderate and severe/death study groups were established. HLA-A, -B, -C, and KIR genotyping were performed using the Lifecodes® HLA-SSO and KIR-SSO kits on the Luminex® 200™ xMAP fluoroanalyser. A probability score using multivariate binary logistic regression analysis was calculated to estimate the likelihood of severe COVID-19. ROC analysis was used to calculate the best cut-off point for predicting a worse clinical outcome with high sensitivity and specificity. A p ≤ 0.05 was considered statistically significant. KIR AA genotype protected positively against severity/death from COVID-19. Furthermore, KIR3DL1, KIR2DL3 and KIR2DS4 genes protected patients from severe forms of COVID-19. KIR Bx genotype, as well as KIR2DL2, KIR2DS2, KIR2DS3 and KIR3DS1 were identified as biomarkers of severe COVID-19. Our logistic regression model, which included clinical and KIR/HLA variables, categorised our cohort of patients as high/low risk for severe COVID-19 disease with high sensitivity and specificity (Se = 94.29%, 95% CI [80.84-99.30]; Sp = 84.55%, 95% CI [79.26-88.94]; OR = 47.58, 95%CI [11.73-193.12], p < 0.0001). These results illustrate an association between KIR/HLA ligand polymorphism and different COVID-19 outcomes and remarks the possibility of use them as a surrogate biomarkers to detect severe patients in possible future infectious outbreaks.

BACKGROUND: This paper outlines a pilot of a new memory aide for breastfeeding conducted in the Northeast of England and North Cumbria between April and August 2023. The United Kingdom has some of the lowest rates of breastfeeding, particularly in the Northeast of England, and as such more needs to be done to support mothers to breastfeed for as long as they would like to. Good support from health professionals can be effective in influencing decisions to breastfeed as well as helping to ensure initiation and continuation of breastfeeding but there is evidence to suggest that professionals and students do not always feel adequately trained and it is here, where memory aides may have value.
METHODS: Key breastfeeding practitioners and educators were brought together to select one of two memory aides for principles of attachment for effective breastfeeding. The selected memory aide, LATCHS, was piloted with 57 participants with a key role in promotion and support of breastfeeding in the Northeast of England and North Cumbria.
RESULTS: Participants conveyed mixed views about the proposed memory aide with more experienced staff reporting more favourable opinions than student midwives and early years practitioners. Experienced staff felt the new memory aide would complement an early memory aide, CHINS, which focused on principles of positioning.
CONCLUSIONS: Findings of the pilot indicate there is a role for a mnemonic to help practitioners understand, recall, and retain theory around attachment for effective breastfeeding and that memory aides can play an important role in complementing existing approaches to education and practice. The participants felt the proposed memory aide had some limitations and suggested important ways for it to be improved, particularly in adding an E to reflect the expecting wording. This produced the final memory aide: LATCHES.
CONCLUSIONS: Using data from the pilot, the memory aide was refined, and the final version LATCHES agreed for wider dissemination. Future research is needed to understand the value of LATCHES on the wider breastfeeding workforce and whether any future improvements can be made to enhance its utility.

BACKGROUND: The chemotherapy regimens recommended for both rhabdomyosarcoma (RMS) and Ewing sarcoma (ES) patients are myelosuppressive and can reduce the absolute neutrophil count (ANC) and subsequently increase the risk of febrile neutropenia (FN). However, only a few studies have focused on the efficacy and safety of granulocyte-colony stimulating factor (G-CSF) drugs in pediatric and adolescent patients with RMS and ES. Our objective was to investigate the efficacy and safety of mecapegfilgrastim, a biosimilar of pegfilgrastim, in prophylaxis of FN for pediatric and adolescent patients with RMS or ES.
METHODS: In this single-arm, single-center, prospective study, pediatric and adolescent patients with RMS or ES were enrolled to receive either VAC (vincristine, cyclophosphamide, dactinomycin) regimen or VDC (vincristine, cyclophosphamide, doxorubicin) regimen in a 3-week cycle, followed by treatment with mecapegfilgrastim (100 μg/kg, maximum 6 mg) given at 24 h after completing chemotherapy. The primary endpoint was the incidence rate of FN. Secondary endpoints included the incidence rate of grade 4 neutropenia, duration of ANC ≤ 0.5 × 109/L, incidence rate of chemotherapy delay or reduction, use of antibiotics, and safety profile.
RESULTS: In total, 2 of the 30 (6.7%, 95% CI: 0.82-22.07) patients experienced FN after the first cycle of chemotherapy. Eight (26.7%, 95% CI: 12.28-45.89) patients experienced grade 4 neutropenia after receiving prophylactic mecapegfilgrastim. Eight patients experienced ANC ≤ 0.5 × 109/L with a median duration of 4.5 days; among them, 6 patients reached the lowest point of their ANC level on day 7, and 5 of them recovered by day 10. No dose reductions, delays, or discontinuation of chemotherapy was reported. Twenty-one (70.0%) patients received antibiotics during the treatment period. No patient experienced FN in the 0-5 years and the 13-18 years groups, and 2 patients experienced FN in the 6-12 years group. Two patients, 6 patients, and no patient experienced grade 4 neutropenia in the 0-5 years, 6-12 years, and 13-18 years groups, respectively.
CONCLUSIONS: Mecapegfilgrastim showed acceptable efficacy and safety profile in pediatric and adolescent patients with RMS or ES. Further randomized studies with large sample size are warranted.
BACKGROUND: This clinical trial was registered at Chictr.org.cn (No.ChiCTR1900022249). Registered on March 31, 2019.

This study aimed to create a Czech questionnaire for pediatric obstructive sleep apnea (POSA) risk screening, a first of its kind in the Czech Republic, where options for child polysomnography are limited. Compiling items from established English questionnaires and supplementing them with additional items, we designed the first version of the Czech questionnaire and tested it in a pilot study with parents of 30 children. After pilot feedback, a revised version with dichotomous and 5-item Likert scale questions was tested on 71 children\'s parents. All children (7-12 years old) underwent a home sleep apnea test to record their apnea-hypopnea index (AHI). The second (40-item) version showed high reliability (93%), with 17 items identified as the most significant. Findings from the final 17-item SEN CZ questionnaire correlated positively with AHI (p < 0.001), demonstrating 84% sensitivity, 86% specificity, and 93% reliability. Three factors, namely breathing problems, inattention, and hyperactivity (characterized by multiple items), were identified to form a higher-order factor of POSA risk, which was further supported by the correlations of their total scores with AHI (p < 0.001). The resulting SEN CZ questionnaire can serve as a tool for POSA risk screening in the Czech Republic without the need to involve medical professionals.

Secondhand vaping exposure is an emerging public health concern that remains understudied. In this study, saliva and exhaled emissions from ENDS users (secondhand) and non-ENDS users (baseline) were collected, firsthand emissions were generated using an automated ENDS aerosol generation system programmed to simulate puffing topography profiles collected from ENDS users. Particulate concentrations and sizes along with volatile organic compounds were characterized. We revealed puffing topography metrics as potential mediators of firsthand and secondhand particle and chemical exposures, as well as metabolic and respiratory health outcomes. Particle deposition modeling revealed that while secondhand emissions displayed smaller deposited mass, total and pulmonary particle deposition fractions were higher than firsthand deposition levels, possibly due to smaller secondhand emission particle diameters. Lastly, untargeted metabolomic profiling of salivary biomarkers of lung injury due to firsthand ENDS exposures revealed potential early indicators of respiratory distress that may also be relevant in bystanders exposed to secondhand vaping scenarios. By leveraging system toxicology, we identified 10 metabolites, including leukotriene D4, that could potentially serve as biomarkers for ENDS use, exposure estimation, and the prediction of vaping-related disease. This study highlights characterization of vaping behavior is an important exposure component in advancing our understanding of potential health effects in ENDS users and bystanders.

Although many neuroimaging studies have evaluated changes in the prefrontal cortex during mindfulness-based interventions, most of these studies were cross-sectional studies of skilled participants or involved pre-post comparisons before and after a single session. While functional near-infrared spectroscopy is a useful tool to capture changes in the hemodynamic response of the prefrontal cortex during continuous mindfulness-based intervention, its ability to detect the accumulated effects of continuous mindfulness-based intervention is currently unclear. We investigated whether a 12-wk online mindfulness-based intervention changed the hemodynamic response of the prefrontal cortex during a verbal fluency task. Eighty-two healthy university students were randomly allocated to a 12-wk online mindfulness-based intervention group or a wait-list control group. The integral values of oxygenated hemoglobin measured using functional near-infrared spectroscopy before and after the intervention were compared to the values in the wait-list group. The intervention condition showed significantly greater functional near-infrared spectroscopy signal activation than the control condition; however, the effect sizes before and after the intervention were small. Thus, continuous mindfulness-based intervention could alter prefrontal cortex function, and functional near-infrared spectroscopy could be useful for measuring the accumulated effects of continuous mindfulness-based interventions. With a better understanding of the association between mindfulness and functional near-infrared spectroscopy signals, functional near-infrared spectroscopy can be used for biofeedback analyses.

Pilot Free Trade Zones (PFTZs) are a crucial new platform for China to build a more open economic system. Existing literature primarily focuses on the \'Bring In\' effect of PFTZs, often overlooking the importance of \'Going Out\' aspects. To bridge this gap, this paper uses data from China\'s Shanghai and Shenzhen A-share listed enterprises from 2007 to 2021 and constructs a time-varying difference-in-differences (DID) model to test the impact of establishment of PFTZs on international expansion of enterprises. The study finds that establishment of PFTZs can significantly promote international expansion of enterprises, with a more pronounced effect on the scope of international expansion than on its depth. Mechanism analysis reveals that PFTZs can facilitate international expansion of enterprises by driving digital transformation, enhancing total factor productivity and management efficiency, and alleviating financing constraints. Notably, senior managers with overseas work experience play a crucial role in enhancing this relationship. Further, PFTZs not only have a linkage effect with the Belt and Road Initiative but also a radiation effect on neighboring cities. This study provides an analytical perspective and empirical evidence for evaluating policy effects of PFTZs and offers valuable insights that will enable PFTZ policies to be refined and facilitate successful implementation of the \'Going Out\' strategy.