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Abstract CRISPR-based gene activation (CRISPRa) is a strategy for upregulating gene expression by targeting promoters or enhancers in a tissue/cell-type specific manner. Here, we describe an experimental framework that combines highly multiplexed perturbations with single-cell RNA sequencing (sc-RNA-seq) to identify cell-type-specific, CRISPRa-responsive cis-regulatory elements and the gene(s) they regulate. Random combinations of many gRNAs are introduced to each of many cells, which are then profiled and partitioned into test and control groups to test for effect(s) of CRISPRa perturbations of both enhancers and promoters on the expression of neighboring genes. Applying this method to a library of 493 gRNAs targeting candidate cis-regulatory elements in both K562 cells and iPSC-derived excitatory neurons, we identify gRNAs capable of specifically upregulating intended target genes and no other neighboring genes within 1 Mb, including gRNAs yielding upregulation of six autism spectrum disorder (ASD) and neurodevelopmental disorder (NDD) risk genes in neurons. A consistent pattern is that the responsiveness of individual enhancers to CRISPRa is restricted by cell type, implying a dependency on either chromatin landscape and/or additional trans-acting factors for successful gene activation. The approach outlined here may facilitate large-scale screens for gRNAs that activate genes in a cell type-specific manner.

Critical reading comprehension is a crucial skill for lifelong learning, but many learners struggle with effectively extracting meaning from texts. This study aimed to address this issue by investigating how a strategy-based instruction intervention could impact learners’ critical reading comprehension competence. The study employed a quantitative approach with a quasi-experimental pre- and post-test control group design. The participants were 155 Grade 9 English first additional language (FAL) learners from two high schools in Gauteng, South Africa. The intervention programme was based on Bloom’s taxonomy categories for learning, teaching and assessing, modified into six strategies: (1) knowledge (pre-reading), (2) comprehension (extracting meaning), (3) application (asking questions), (4) analysis (reflection), (5) evaluation (outlining key concepts and decision-making) and (6) synthesis (summarising). Using a two-way mixed analysis of variance (ANOVA) paired sample t-tests, the study found that while both groups’ critical reading comprehension competence increased, the experimental group that received the strategy-based instruction intervention performed significantly better in the post-test, showing a significant improvement compared to the control group. The findings demonstrated that providing clear explanations, instruction and support on reading strategies can enhance learners’ critical reading comprehension, highlighting the mutual reliance between teaching and learning. The teacher’s instructional approach in guiding learners to effectively use reading strategies is crucial for enhancing critical reading comprehension in the English FAL classroom. In conclusion, the results suggest that the intervention effectively enhanced critical reading comprehension competence, and if implemented by teachers, it could potentially help break the cycle of poor critical reading comprehension among English FAL learners. Contribution: The contribution of this study lies in its provision of empirically supported perspectives and insights into the effectiveness of strategy-based instruction interventions. It emphasises the necessity of employing explicit reading comprehension pedagogies and targeted interventions, along with explicit instructional strategies, to address the reading comprehension challenges encountered by English FAL learners in South Africa and beyond.

Abstract Cassava brown streak disease (CBSD) caused by Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV) is the most economically important viral disease of cassava. As cassava is a vegetatively propagated crop, the development of rapid and sensitive diagnostics would aid in the identification of virus-free planting material and development of effective management strategies. In this study, a rapid, specific and sensitive real-time reverse transcription recombinase polymerase amplification (RT-RPA) assay was developed for real-time detection of CBSV and UCBSV. The RT-RPA was able to detect as little as 2 pg/µl of purified RNA obtained from infected cassava leaves, a sensitivity equivalent to that obtained by quantitative real-time reverse transcription PCR (qRT-PCR), within 20 min at 37 °C. Further, the RT-RPA detected each target virus directly from crude leaf and stem extracts, avoiding the tedious and costly isolation of high-quality RNA. The developed RT-RPA assay provides a valuable diagnostic tool that can be adopted by cassava seed certification and virus resistance breeding programs to ensure distribution of virus-free cassava planting materials to farmers. This is the first report on the development and validation of crude sap-based RT-RPA assay for the detection of cassava brown streak viruses (UCBSV and CBSV) infection in cassava plants.

Abstract Glioblastoma (GBM) is a highly lethal type of cancer. GBM recurrence following chemoradiation is typically attributed to the regrowth of invasive and resistant cells. Therefore, there is a pressing need to gain a deeper understanding of the mechanisms underlying GBM resistance to chemoradiation and its ability to infiltrate. Using a combination of transcriptomic, proteomic, and phosphoproteomic analyses, longitudinal imaging, organotypic cultures, functional assays, animal studies, and clinical data analyses, we demonstrate that chemoradiation and brain vasculature induce cell transition to a functional state named VC-Resist (vessel co-opting and resistant cell state). This cell state is midway along the transcriptomic axis between proneural and mesenchymal GBM cells and is closer to the AC/MES1-like state. VC-Resist GBM cells are highly vessel co-opting, allowing significant infiltration into the surrounding brain tissue and homing to the perivascular niche, which in turn induces even more VC-Resist transition. The molecular and functional characteristics of this FGFR1-YAP1-dependent GBM cell state, including resistance to DNA damage, enrichment in the G2M phase, and induction of senescence/stemness pathways, contribute to its enhanced resistance to chemoradiation. These findings demonstrate how vessel co-option, perivascular niche, and GBM cell plasticity jointly drive resistance to therapy during GBM recurrence.

Abstract Background In the face of drought and food insecurity emergency, evidence on access to health and nutrition services is important. Karamoja is one of the regions that have experienced extreme drought and food insecurity emergency in Uganda. As a part of the drought and food insecurity emergency response, World Health Organization (WHO) with Ministry of Health (MoH) has designed and implemented a qualitative study in 15 districts that have experienced drought and food insecurity emergency in north-east Uganda. Thus, we aimed to explore the barriers of access to health and nutrition services in drought and food insecurity emergency affected districts in north-east Uganda. Methods We employed a descriptive qualitative study design. We interviewed 30 patients and 20 Village Health Teams (VHT) from 15 districts. We employed an in-depth interview with semi-structured questions to collect data until information saturation reached. We used thematic data analysis approach by ATLAS.ti version 7.5.1.8 software. Results Of the 30 interviewed subjects, 15 were female, and the median age of the subjects was 29 years with interquartile range (IQR) of 23 to 37 years. Majority (68.8%) of subjects reported that access to health and nutrition services was harder to them. Four themes: sociocultural and economic; environmental; health system, and individual related factors were identified as the barriers of access to health and nutrition services. Conclusion The present study identified several modifiable barriers that hinder access to health and nutrition services in drought and food insecurity affected districts. Comprehensive interventions aimed at addressing sociocultural, economic, environmental, health system and subject related challenges are required to improve access to health and nutrition services in drought and food insecurity affected setups.

Abstract Background Essential health services can be disrupted due to several naturally occurring public health emergencies such as drought, flood, earthquake and outbreak of infectious diseases. However, little evidence exists on the status of essential health services delivery under the effect of drought and food insecurity. North-east Uganda is severely affected by prolonged drought that significantly affected the livelihood of the residents. Therefore, we aimed to determine the current status of essential health services and quality improvement (QI) actions in health facilities in north-east Uganda. Methods We used a descriptive cross-sectional study design to assess the availability of essential health service and quality improvement activities in drought and food insecurity affected districts of north-east Uganda. We included a total of 150 health facilities from 15 districts with proportionated multistage sampling method. We interviewed health facilities’ managers and services focal persons using structured questionnaire and observation checklist. We used a descriptive statistic to analyze the data with SPSS version 22. Results A few health facilities (8.7%) had mental health specialist. There was also lack of capacity building training on essential health services. Considerable proportion of health facilities had no non-communicable diseases (38.3%), mental health (47.0%), and basic emergency obstetric care (40.3%) services. Stock out of essential medicines were observed in 20% of health facilities. There was lack of supportive supervision, and poor documentation of QI activities. Conclusion Essential health service and QI were suboptimal in drought and food insecure emergency affected districts. Human resource deployment (especially mental health specialist), provision of capacity building training, improving non-communicable diseases, mental health and basic emergency obstetric care services are required to improve availability of essential health services. Supporting supply chain management to minimize stock out of medicines, and promoting QI activities are also vital to assure quality of health service in drought and food insecurity affected districts in north-Eastern Uganda.

The rapid development of technology has led to the emergence of innovative teaching approaches, such as high-fidelity patient simulation (HFPS). HFPSs have been shown to significantly enhance students’ decision-making and intellectual skills. This study aimed to investigate how students learn from the pre-briefing to observation period of the facilitation of the HFPS based on the original quasi-experimental studies. This study analyzed the narratives from 92 students in the intervention group about their learning during the pre-briefing and observation of facilitation. The results indicated that the students learned more independently, received better support and resources for learning, were provided with more appropriate and safe care for the simulated case, and developed higher-level intellectual skills, such as self-directed learning, critical thinking, and clinical reasoning. Importantly, the structured guidelines provided roles and responsibilities and guiding questions or aspects for observation that directed the students to learn more actively and effectively while performing their roles in the HFPS. The structured guidelines serve as a roadmap to instruct students on learning during pre-briefing and applying what they have learned during the facilitation of the HFPS. This roadmap includes the learning journey from novice to competence in knowledge and skills and also from knowing to application. Therefore, this study’s results have contributed important knowledge about well-structured HFPS guidelines for all stages of the HFPS, addressing the need for adequate guidance and learning support during the pre-briefing and observation of facilitation. The elements identified during the pre-briefing and observation of facilitation are crucial for directing students to learn and significantly enhance their understanding and application of knowledge and skills, ultimately promoting the development of higher-level intellectual skills, professionalism, and engagement. Nurse educators can incorporate these elements into HFPS training in curricula to enhance students’ involvement and optimize the HFPS as an effective teaching tool with structured guidelines providing guidance and support throughout various stages of HFPS training.

Purpose Pregnancy and the postpartum period are increasingly recognised as sensitive windows for cardiometabolic disease risk. Growing evidence suggests environmental exposures, including endocrine-disrupting chemicals (EDCs), are associated with an increased risk of pregnancy complications that are associated with long-term cardiometabolic risk. However, the impact of perinatal EDC exposure on subsequent cardiometabolic risk post-pregnancy is less understood. The Environmental Reproductive and Glucose Outcomes (ERGO) Study was established to investigate the associations of environmental exposures during the perinatal period with post-pregnancy parental cardiometabolic health.Participants Pregnant individuals aged ≥18 years without pre-existing diabetes were recruited at

No abstract available.

Background: The importance of oral health in older adults, especially those in long-term care institutions (LTCIs), has been widely recognized. This study aimed to evaluate the sustainability of an oral health educational program (OHEP) for healthcare providers by measuring changes in their knowledge, attitudes, and practice (KAP) towards oral care provision 3 and 6 months after completing the OHEP. Methods: A pragmatic direct care nursing education trial with a control group was conducted to evaluate the sustainability of an OHEP by examining changes in KAP 3 and 6 months after the OHEP. The OHEP comprised both knowledge and skills related to oral care, whereas the control group received standard support in accordance with usual oral care practice. Results: The study included 20 healthcare providers in the intervention group and 20 in the control group. At 6 months post-OHEP, a significant difference in knowledge was observed between the two groups, with the intervention group maintaining a positive effect (mean 13.90). Conversely, the control group showed a significant decline in knowledge (from mean 14.25 to 12.10). Both groups showed an improvement in attitudes regarding oral health, with the intervention group exhibiting better results 3 months post-OHEP. Intervention group participants rated oral care as a higher priority. Conclusions: An OHEP program for LTCI direct care staff provides enhanced knowledge and attitudes toward oral health care. Regular training in direct care and additional support may be needed to sustain optimal effects on oral care practice.

The coronaviral pandemic has led to a shift in traditional teaching methods to more innovative approaches, such as high-fidelity patient simulation (HFPS), which can improve students’ clinical judgment and decision making for quality patient care. A modified guideline was introduced to enhance students’ satisfaction and self-confidence in learning through HFPS. The study involved 189 baccalaureate nursing students, with 92 in the intervention group and 97 in the control group. The intervention group received the modified HFPS guideline, while the control group received standard treatment with basic instruction. After the HFPS debriefing session, students provided narrative feedback on their learning experiences. The quantitative results showed that students in the intervention group reported a significant improvement in satisfaction and self-confidence in learning compared to the control group. The modified HFPS guideline provided clear guidance for students to learn and apply knowledge and skills more effectively, leading to increased engagement during interactive simulation sessions. The results suggest that the HFPS guideline should be added to the curriculum to enhance students’ satisfaction and self-confidence in learning, even for junior students. After the pandemic, innovative teaching methods, such as HFPS, can be necessary and beneficial for healthcare professional training.

Background: In the education sector, technologies are (or can be) repurposed in the Fourth Industrial Revolution (4IR) to facilitate teaching and learning. However, this shift may impact on learners’ access to reading materials and reading motivation. Objectives: The aim of the study was to determine whether Grade 8 English First Additional Language (FAL) learners’ motivation to read is influenced by access to reading materials in a traditional classroom setting or through online reading platforms. Method: Likert-scale survey questionnaires were used to measure this relationship. Data from 466 Grade 8 English FAL learners from six South African township schools were collected and SPSS version 25 was used to analyse the data. Results: In traditional classrooms, the findings showed that increased access positively influenced motivation, whereas this effect was not evident in online reading platforms. However, in both settings, motivation increased when learners had access to a diverse selection of reading materials. Finally, the findings indicated a parallel relationship between learners’ low motivation and limited access to reading platforms. Conclusion: The study concluded that while traditional classrooms continue to play an important role in global education, the ongoing evolution of technology-based learning platforms has compelled a shift in reading practices toward online platforms, which are reshaping the educational landscape, demonstrating the increasing significance of technology-mediated learning in modern education. Contribution: The authors recommend enhanced collaboration between teachers and learners to devise innovative methods for motivating online reading, ensuring learners’ inclusion in the 4IR literacy pedagogy evolution.

Abstract Functional recovery following incomplete spinal cord injury (SCI) depends on the rewiring of motor circuits during which supraspinal connections form new contacts onto spinal relay neurons. We have recently identified a critical role of the presynaptic organizer FGF22 for the formation of new synapses in the remodeling spinal cord. Here, we now explore whether and how targeted overexpression of FGF22 can be used to mitigate the severe functional consequences of SCI. By targeting FGF22 expression to either long propriospinal neurons, excitatory interneurons, or a broader population of interneurons, we establish that FGF22 can enhance neuronal rewiring both in a circuit‐specific and comprehensive way. We can further demonstrate that the latter approach can restore functional recovery when applied either on the day of the lesion or within 24 h. Our study thus establishes viral gene transfer of FGF22 as a new synaptogenic treatment for SCI and defines a critical therapeutic window for its application.

Purpose: The purpose of this study is to examine the relationship between telehealth use, telehealth satisfaction, and chronic medical conditions among residents living in Appalachian and non-Appalachian communities. Sample: A COVID-19 public health survey was distributed via social media and healthcare clinics in the tri-state region of central Appalachia. Survey responses were limited to adults aged ≥18 years who consented to participate in the survey that self-identified as an individual with one or more chronic medical conditions (n=195). Method: Simple descriptive statistics including frequencies, percentages, means, and standard deviations (SDs) were calculated for variables of interest both overall and by subgroups of interest. Chi-squared tests were used to compare categorical outcomes between groups of interest, while two-sample t-tests were used for continuous outcomes. Significance for all tests was determined using an α level of 0.05. Findings: There is no statistically significant relationship between respondents with regard to using telehealth services, satisfaction rates related to telehealth use, or reasons for electing not to use telehealth services during the COVID-19 pandemic. However, there was a trending statistical relationship between county status and the use of telehealth services in Appalachia with those counties doing economically better being more likely to use telehealth services as compared to those fairing less well (p=0.053). Findings also suggest that people living in urban areas of Appalachia were more likely to be satisfied using telehealth services than those living in non-urban areas of Appalachia (p=0.01). Conclusions: Research is still limited as to how the expansion of broadband capabilities during the COVID-19 pandemic has benefited those residing in Appalachia in terms of managing chronic health conditions. Future research should focus on expanding participation among Appalachian respondents looking for specific differences related to location within Appalachia, age, gender, ethnicity, and socioeconomic status. Keywords: Telehealth, Satisfaction, Rural, Appalachian, COVID-19 DOI: https://doi.org/10.14574/ojrnhc.v22i2.715

The field of sleep research is both broad and rapidly evolving. It spans from the diagnosis of sleep-related disorders to investigations of how sleep supports memory consolidation. The study of sleep includes a variety of approaches, starting with the sole focus on the visual interpretation of polysomnography characteristics and extending to the emergent use of advanced signal processing tools. Insights gained using artificial intelligence (AI) are rapidly reshaping the understanding of sleep-related disorders, enabling new approaches to basic neuroscientific studies. In this opinion article, we explore the emergent role of AI in sleep research, along two different axes: one clinical and one fundamental. In clinical research, we emphasize the use of AI for automated sleep scoring, diagnosing sleep-wake disorders and assessing measurements from wearable devices. In fundamental research, we highlight the use of AI to better understand the functional role of sleep in consolidating memories. While AI is likely to facilitate new advances in the field of sleep research, we also address challenges, such as bridging the gap between AI innovation and the clinic and mitigating inherent biases in AI models. AI has already contributed to major advances in the field of sleep research, and mindful deployment has the potential to enable further progress in the understanding of the neuropsychological benefits and functions of sleep.

Poor oral health is a growing concern among older populations. It is often caused by a failure to maintain proper oral hygiene and inaccessible dental care. Poor oral health in older individuals in long-term care institutions (LTCIs) can be attributed to the fact that healthcare workers might be poorly trained in oral care assessment and practice. To address this issue, an assessment tool has been developed and validated to guide and evaluate healthcare workers’ oral care practices, ensuring the delivery of adequate care and early detection of dental diseases in LTCIs. The tool includes an oral health assessment and an assessment of oral care procedures. It was developed following a robust literature review, two stages of expert reviews, content validity checks, and a pilot study. A total of twenty-three items were developed and validated, with seven items related to oral health assessment and sixteen related to oral care procedures. The items were assessed for content validity and relevance, with high values of 1 obtained for all Item-level Content Validity Index (I-CVI), Scale-level Content Validity Index (S-CVI), and S-CVI/Universal Agreement (UA) scores. This indicates a high level of agreement among the experts (n = 12) regarding the relevance and importance of the items. A pilot study involving 20 nursing students confirmed the tool’s reliability, applicability, and feasibility, demonstrating its high appropriateness and applicability. The newly developed and validated assessment tool can effectively guide and evaluate healthcare workers’ oral care practices, enhancing their competence and improving the oral health of older residents.

Background: Much attention has been paid to advocate proper oral care/hygiene provision by healthcare providers in long-term care institutions (LTCIs). This study aimed to evaluate the effects of an oral health education (OHE) programme (intervention) on knowledge, attitudes, and practice (KAP) of healthcare providers in providing oral care/hygiene to older residents in LTCIs. Methods: A case control study was conducted at two LTCIs, with one assigned as the intervention group and the other as the control group. A KAP survey was administered before and after the intervention, and oral status was assessed by standardized clinical photographs taken before and after oral hygiene provision on three older residents. Results: A total of 40 healthcare providers (20 in intervention and 20 in control groups) participated, with the attitudes and overall KAP significantly improved in the intervention group after the OHE programme. Interestingly, the knowledge of those in the control LTCI was significantly declined at re-evaluation (mean scores were from 17.25 to 14.30), indicating inadequate oral health and care training despite having more experience in taking care of older people. Significant differences in practice were observed between the two groups after the OHE programme (p = 0.006). The three older residents exhibited poor oral health and multiple oral problems. Conclusions: This study revealed that the OHE programme effectively improved attitudes of the healthcare providers and provided a sustaining effect on attitude towards oral health and oral care. However, there were still inadequacies in oral hygiene provision by some healthcare providers, possibly due to unattended oral diseases and hygiene needs, as well as personal and environmental barriers that merit further investigation. Regular evaluation and enforcement of oral care/hygiene provision in LTCIs are necessary to maintain oral health and prevent dental and gum diseases in older residents. Immediate referral for dental treatment is recommended for older people with signs of dental/oral disease(s).

Reservoirs are human-made ecosystems with diverse purposes that benefit humans both directly and indirectly. They however cause changes in geomorphological processes such as sediment cycling and influence the composition and structure of aquatic biota. This study aimed to identify water and sediment quality parameters as drivers of macroinvertebrates and fish communities during the cool-dry and hot-wet seasons in the littoral zones of three subtropical reservoirs (Albasini, Thathe and Nandoni). Macroinvertebrates and fish were collected from three sites (n = 3 from each site) in each reservoir. A total of 501 and 359 macroinvertebrates and fish individuals were collected throughout the sampling period, respectively. The present study employed a two-way ANOVA in conjunction with redundancy analysis (RDA) to assess the relationships that exist between water and sediment variables, macroinvertebrates diversity and species abundances across seasons. Based on the two-way ANOVA model, significant differences were observed across reservoirs for evenness, Simpson’s diversity, and total abundance, while seasonal differences were observed for most metrics, with exception for evenness. The RDA results identified four water variables (i.e., water temperature, oxidation–reduction potential, pH and conductivity) and one sediment metal (Mg) as the most important parameters in driving the fish community structure. Field observations and metal results attest that the Nandoni reservoir shows high concentrations of metals in sediments as compared to other reservoirs, suggesting that anthropogenic activities such as car washing, brick making, recreation, fishing, wastewater treatment work and landfill site may be the major contributor of metals to the Nandoni reservoir, which accumulate in the littoral zones. Findings of this study highlight the need to analyze reservoir ecological conditions at several scales. The study of macroinvertebrates and fish, water, and sediment chemistry in the littoral zone laid the groundwork for proposing measures for conserving aquatic ecosystems.

Abstract Background We have a limited understanding on how to best integrate technologies to support antiretroviral therapy (ART) adherence in routine HIV care. Methods We conducted semi-structured interviews with multidisciplinary providers caring for pregnant and postpartum people with HIV and asked providers about their perspectives on utilizing adherence support technologies such as text messages, video check-ins with providers or automated with facial recognition for directly-observed-therapy, signaling pill bottle, and signaling pill to support ART adherence. Each approach generated an adherence report. The interview instrument was guided by the Consolidated Framework for Implementation Research and included questions on the implementation climate, barriers, and facilitators to the clinical integration of the adherence approach and strategies that could be used to maximize this integration. The order of adherence support technologies was randomized to minimize bias. We used a modified grounded theory to develop the coding structure and two coders applied the codebook to the transcripts after establishing strong inter-rater reliability with 20% of interviews (kappa = 0.82). Results Between March and December 2020, we conducted 26 in-depth, semi-structured interviews with providers who weighed several factors when considering each approach, including the approach’s effect on patient-provider interaction in and outside of the clinic visit, timing for and duration of the approach’s utility, threat of disclosing status, and added burden to providers (e.g., needing to act on generated information) or to patients (e.g., needing to hide the signaling pills, responding to text messages). Providers’ most preferred approach was text-messages, and the least preferred was the signaling pill. Barriers to acceptability varied by approach and included perceived surveillance, violation of privacy, added time demand for providers, potential inaccuracy of the adherence data generated, and negative impact on the patient-provider relationship, particularly if the approach was perceived as coercive. Payers anticipated regulatory hurdles with unfamiliar approaches, particularly the signaling pill and signaling pill bottle. Facilitators included strengthened therapeutic alliance, predictable reminder mechanisms, and options for customization according to patient preference. Conclusions Our study elucidates barriers and facilitators to integrating technology-based adherence support approaches in clinical care to support adherence of pregnant and postpartum people with HIV.

Summary: The piRNA pathway in mosquitoes differs substantially from other model organisms, with an expanded PIWI gene family and functions in antiviral defense. Here, we define core piRNA clusters as genomic loci that show ubiquitous piRNA expression in both somatic and germline tissues. These core piRNA clusters are enriched for non-retroviral endogenous viral elements (nrEVEs) in antisense orientation and depend on key biogenesis factors, Veneno, Tejas, Yb, and Shutdown. Combined transcriptome and chromatin state analyses identify transcriptional readthrough as a conserved mechanism for cluster-derived piRNA biogenesis in the vector mosquitoes Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Anopheles gambiae. Comparative analyses between the two Aedes species suggest that piRNA clusters function as traps for nrEVEs, allowing adaptation to environmental challenges such as virus infection. Our systematic transcriptome and chromatin state analyses lay the foundation for studies of gene regulation, genome evolution, and piRNA function in these important vector species.

Which contralesional circuits adapt after traumatic brain injury (TBI) is unclear. Here the authors used in vivo imaging, retrograde labeling, rabies tracing, clearing and functional imaging to demonstrate that callosal neurons selectively adapt after TBI in mice.

Presenting ALMA toolbox, an open source Python repository for automatic analysis of mouse locomotion using bodypart coordinates from markerless pose estimation tools. ALMA is capable of analyzing both healthy and CNS-injured mice. ALMA is also capable of predicting onset of disease in a multiple sclerosis model.

Serotonin is a monoamine neurotransmitter synthetized in various populations of brainstem neurons. In the spinal cord, descending serotonergic projections regulate postural muscle tone, locomotion and rhythm and coordination of movements via the Central Pattern Generator. Following a spinal cord injury, serotonergic projections to the lumbar spinal cord, where the Central Pattern Generators are located, are interrupted resulting in devastating locomotor impairments and changes in the expression and activation of serotonin and its spinal receptors. The molecular cues that control the precise patterning and targeting of serotonergic inputs onto Central Pattern Generator networks in healthy animals or after injury are still unknown. In our recent research work, we have been particularly interested in Semaphorin7A, which belongs to the Semaphorins family involved in guiding growing axons and controlling plasticity of synaptic connections. In this review, we discuss the role of Semaphorin7A signaling as an important molecular actor that instructs the patterning of serotonin inputs to spinal Central Pattern Generator networks. We show that Semaphorin7A controls the wiring of descending serotonin axons in the spinal cord. Our results reveal that mistargetting of serotonin fibers in the spinal cord is compensated in healthy uninjured Semaphorin7A deficient mice so that their gross locomotion proceeds accurately. We also demonstrate that when the system is challenged with a spinal lesion, the pattern of post-injury serotonin expression is significantly altered in Semaphorin7A deficient mice with specific ectopic targeting of serotonin fibers in the lumbar spinal cord. Compensatory mechanisms in place in uninjured Semaphorin7A deficient mice are lost and injured Semaphorin7A deficient mice exhibit a worsening of their post-injury locomotor abilities. Our findings identify Semaphorin7A as a critical determinant of serotonergic circuit formation in healthy or spinal cord injured mice.

AIM: To evaluate the graft rejection and visual outcomes after penetrating keratoplasty (PK) in the presence of various congenital corneal opacities in children. METHODS: In this retrospective cohort study, children who underwent PK were then followed for 5y. The patient's medical records were collected from June 2014 until June 2019 and analyzed in December 2019. All patients were children under three years old with congenital corneal opacities with or without microcornea who came to a pediatric ophthalmologist and underwent PK in Jakarta Eye Center (JEC). Beforehand, all children have participated in a thorough evaluation for PK. In the case of severe microcornea was not advised to undergo surgery. The visual outcomes and graft survival rate were described in percentages. The graft survival plot was presented with Kaplan-Meier, while the visual acuity was analyzed using the Wilcoxon signed ranks test. RESULTS: Sixteen eyes from eleven patients (seven girls and four boys) underwent PK. The graft survival rate of the first 6, 12, and 18 mo later of keratoplasty was 100%, 83.3%, and 66.7%, respectively. The overall mean survival time is 22mo (standard error 2.419), and no significant difference between the patients underwent PK before and after 36mo of their age (P=0.52). The graft failure was 50%, and post-surgery complications included cataract 43.7%, band keratopathy 12.5%, and scleromalasia 6.25%. Wilcoxon test analysis of visual acuity post keratoplasty was not statistically significant (P=0.34), while overall showed 44% improvements of visual outcome for 5y of follow-up. With a good survival at one year up to 22mo (83.3%), the visual acuity could be achieved (63%), and showed improvements (44%) during follow-up. CONCLUSION: The complications are frequent for pediatric PK. Thus, corneal surgery on infants requires careful case selection, adequate pre-operative evaluation, skilled surgery (optical correction), very close cooperation family–physician, intensive post-operation care, and amblyopia management in the future.

The sterilization of medical devices is paramount to achieve an acceptable level of sterility assurance and to prevent hospital-acquired infections. However, some medical devices cannot be sterilized by usual processes such as autoclave (AC) and gamma-ray irradiation (GI). A new non-thermal plasma (NTP) process using sealed bag that preserves the sterile state of the devices could be used as an alternative sterilization method. The aim of the study was to assess the cytocompatibility of titanium and poly(etheretherketone) (PEEK) surfaces after O2-NTP sterilization compared to GI and AC. MG-63 osteoblast-like cells were seeded on titanium (TA6V) and PEEK disks sterilized by AC, GI and O2-NTP. The cells’ viability and proliferation, determined by WST-1 and DNA quantification respectively, were enhanced whatever the material types from 3 to 10 days. When seeded on titanium, MG-63 cells showed a higher viability and proliferation after GI and O2-NTP treatment compared to AC treatment. When cultured on PEEK, MG-63 cells showed a higher viability after O2-NTP treatment. No difference of proliferation was observed whatever the sterilization processes. The cell colonization of the materials’ surface was confirmed by scanning electron microscopy. Lactate dehydrogenase (LDH) assay revealed no cytotoxicity. Thus, O2-NTP led to similar cell responses to AC and GI and could be a cost-effective alternative process to the usual sterilization methods for fragile medical devices.

The rat model can be used to assess ileal protein digestibility rapidly and in first intention, but no standardised method exists. Our objective was to compare methods to assess protein digestibility, depending on collection site (ileum/caecum) and use of a non-absorbable marker. A meal containing either casein, gluten or pea protein and chromium oxide as non-absorbable marker was given to male Wistar rats and the entire digestive content was collected 6 h later. Total chromium recovery was incomplete and variable, depending on protein source. We observed no significant difference in digestibility between the methods for any of the protein sources tested. Although none of the methods tested is optimal, our results suggest that caecal digestibility can be used as a proxy of ileal digestibility in rats without using a non-absorbable marker. This simple method makes it possible to evaluate protein digestibility of new alternative protein sources for human consumption.

The mechanisms by which early microbial colonizers of the neonate influence gut development are poorly understood. Bacterial bile salt hydrolase (BSH) acts as a putative colonization factor that influences bile acid signatures and microbe-host signaling pathways and we considered whether this activity can influence infant gut development. In silico analysis of the human neonatal gut metagenome confirmed that BSH enzyme sequences are present as early as one day postpartum. Gastrointestinal delivery of cloned BSH to immature gnotobiotic mice accelerated shortening of the colon and regularized gene expression profiles, with monocolonised mice more closely resembling conventionally raised animals. In situ expression of BSH decreased markers of cell proliferation (Ki67, Hes2 and Ascl2) and strongly increased expression of ALPI, a marker of cell differentiation and barrier function. These data suggest an evolutionary paradigm whereby microbial BSH activity potentially influences bacterial colonization and in-turn benefits host gastrointestinal maturation.

Abstract Coronavirus disease 2019 (COVID-19) pandemic is rapidly evolving and is a serious public health threat worldwide. Timely and effective control of the pandemic is highly dependent on preventive approaches. Perception of risk is a major determinant of health behavior. The current study explores the association between actual risk and perceived risk for one’s self, family/friends and friends, and community. A questionnaire was administered to participants in Central Appalachia (n = 102). The actual risk was based on the number of chronic conditions of the following conditions: hypertension, heart disease, cancer, diabetes, and chronic obstructive pulmonary disease. Participants were also queried about their perception of risk for COVID-19. Generalized Linear Models were used to independently evaluate the likelihood of perceived risk for one’s: self, family/friends, and community, based on actual risk. Actual risk for COVID-19 was significantly associated with higher likelihood of higher perception of risk for one’s self (b = 0.24; p = 0.04), but not with one’s family/friends (b = 0.05; p = 0.68), or one’s community (b = 0.14; p = 0.16). No health insurance was negatively associated with perception of risk for self (b = −0.59; p = 0.04) and family/friends (b = −0.92; p

The world’s population is growing continually and is projected to reach nine billion by the year 2050. This growth rate requires increased and economically viable food production and an adequate supply of quality water to sustain life. Increased food production and supply of water require adding fertilizers and possible recycling of wastewater, to address the improvement of soils’ nutritional status and potable water shortages, respectively. The objectives of this work were to determine the nutrients in sewage-impacted wastewater, borehole water, agricultural waste, and commercial fertilizer (control) materials, and their heavy metal content was also carried out to determine their suitability for use. In addition, Moringa seed pods and Morula nutshells were investigated as a bioremedial approach for the removal of toxic metals from aqueous samples. An attempt to regenerate sorbent was made since the saturated sorbents that contain the metal ions are not safe for disposal as they can pollute the environment. Nutrients were analyzed by HPLC, while metals were analyzed using a Varian 220FS Atomic Absorption Spectrometer operated with air/acetylene. Nonedible agricultural materials were found to contain appreciable amounts of plant nutrients such as nitrates (NO3-), nitrites (NO2-), and phosphates (PO43-) as well as metal ions such as magnesium, copper, and zinc, which are beneficial for plant growth. Results obtained from analysis of sewage water effluent showed that heavy metal and nutrient concentrations decreased in the treatment stage. The utilization of Moringa oleifera seed pods for metal removal from wastewater is viable and would reduce costs for waste disposal and can offer alternatives to conventional methods for the removal of unwanted or toxic species from the environment. It showed potential for removing selected metal ions such as Pb, Cd, Cu, Fe, and Zn from polluted water. This organically treated wastewater is environmentally friendly and may be used for applications which do not require potable water, such as irrigating golf courses, lawns, and crops, or for industrial purposes, if proper measures are taken to ensure its quality.

High-fidelity patient simulation (HFPS) is widely used in professional training to enhance students’ competence in clinical management. A guideline for HFPS provides a systematic approach to direct students to learning during the simulation process. Problem-solving (PS) and clinical reasoning (CR) skills are essential to developing students’ professional competence in safe and effective care. These two skills should be initiated in the early training. A structured guideline was developed for HFPS. This study aimed to investigate the effects of the structured HFPS guideline on the development of PS and CR skills in junior nursing students. The students were required to go through four sessions, pre-briefing, simulation design, facilitation, and debriefing, for the HFPS; the study utilized the Problem-Solving Inventory (PSI) and the Nurses’ Clinical Reasoning Scale (NCRS) to measure PS and CR abilities before and after HFPS. Bivariate analysis, a one-sample t-test, and an independent t-test were performed to evaluate the performance of the PS and CR skills during the two study periods. A total of 189 students were recruited, with 92 in the intervention group and 97 in the control group. The research assistant was responsible for student recruitment through email invitations and allocating the students into the control group or the intervention group. A Wilcoxon analysis was performed and revealed significant differences in PS and CR between the two groups (p < 0.001). The analytic results showed that the PSI, particularly in domains of Problem-Solving Confidence (PSC) (p < 0.001) and overall PS (p < 0.001), and the CR (p < 0.001) had significant improvement after HFPS, particularly in the intervention group. The study concluded that the structured HFPS guideline significantly improved the students’ problem-solving and clinical reasoning abilities. Nurse educators play an important role in providing explicit learning instructions in a simulation guideline that directs and guides students to learn at each stage of HFPS. The students can be directed to be engaged in their learning through HFPS to enhance their competence in knowledge and skill development (PS and CR) for their personal and professional development.

Sonic Hedgehog medulloblastoma (Shh-MB) comprises four subtypes each with distinct clinical traits. Here the authors characterize the genome, transcriptome, and methylome of Shh-MB subtypes, revealing a complex fusion landscape and the molecular convergence of MYCN and cAMP signaling pathways.

The arginine methyltransferase PRMT5 is over-expressed in cancer and has a role in the maintenance of stem cells. Here, the authors show that PRMT5 inhibitors can block the growth of patient derived glioblastoma stem cell cultures in vitro and in vivo, suggesting that PRMT5 inhibition may be a useful therapeutic strategy