ADAR regulates APOL1 via A-to-I RNA editing by inhibition of MDA5 activation in a paradoxical biological circuit.

APOL1 risk variants are associated with increased risk of kidney disease in patients of African ancestry, but not all individuals with the APOL1 high-risk genotype develop kidney disease. As APOL1 gene expression correlates closely with the degree of kidney cell injury in both cell and animal models, the mechanisms regulating APOL1 expression may be critical determinants of risk allele penetrance. The APOL1 messenger RNA includes Alu elements at the 3' untranslated region that can form a double-stranded RNA structure (Alu-dsRNA) susceptible to posttranscriptional adenosine deaminase acting on RNA (ADAR)-mediated adenosine-to-inosine (A-to-I) editing, potentially impacting gene expression. We studied the effects of ADAR expression and A-to-I editing on APOL1 levels in podocytes, human kidney tissue, and a transgenic APOL1 mouse model. In interferon-γ (IFN-γ)-stimulated human podocytes, ADAR down-regulates APOL1 by preventing melanoma differentiation-associated protein 5 (MDA5) recognition of dsRNA and the subsequent type I interferon (IFN-I) response. Knockdown experiments showed that recognition of APOL1 messenger RNA itself is an important contributor to the MDA5-driven IFN-I response. Mathematical modeling suggests that the IFN-ADAR-APOL1 network functions as an incoherent feed-forward loop, a biological circuit capable of generating fast, transient responses to stimuli. Glomeruli from human kidney biopsies exhibited widespread editing of APOL1 Alu-dsRNA, while the transgenic mouse model closely replicated the edited sites in humans. APOL1 expression in mice was inversely correlated with Adar1 expression under IFN-γ stimuli, supporting the idea that ADAR regulates APOL1 levels in vivo. ADAR-mediated A-to-I editing is an important regulator of APOL1 expression that could impact both penetrance and severity of APOL1-associated kidney disease.

Mycobacterial RNase E cleaves with a distinct sequence preference and controls the degradation rates of most Mycolicibacterium smegmatis mRNAs.

The mechanisms and regulation of RNA degradation in mycobacteria have been subject to increased interest following the identification of interplay between RNA metabolism and drug resistance. Mycobacteria encode multiple ribonucleases predicted to participate in mRNA degradation and/or processing of stable RNAs. RNase E is hypothesized to play a major role in mRNA degradation because of its essentiality in mycobacteria and its role in mRNA degradation in gram-negative bacteria. Here, we defined the impact of RNase E on mRNA degradation rates transcriptome-wide in the nonpathogenic model Mycolicibacterium smegmatis. RNase E played a rate-limiting role in degradation of the transcripts encoded by at least 89% of protein-coding genes, with leadered transcripts often being more affected by RNase E repression than leaderless transcripts. There was an apparent global slowing of transcription in response to knockdown of RNase E, suggesting that M. smegmatis regulates transcription in responses to changes in mRNA degradation. This compensation was incomplete, as the abundance of most transcripts increased upon RNase E knockdown. We assessed the sequence preferences for cleavage by RNase E transcriptome-wide in M. smegmatis and Mycobacterium tuberculosis and found a consistent bias for cleavage in C-rich regions. Purified RNase E had a clear preference for cleavage immediately upstream of cytidines, distinct from the sequence preferences of RNase E in gram-negative bacteria. We furthermore report a high-resolution map of mRNA cleavage sites in M. tuberculosis, which occur primarily within the RNase E-preferred sequence context, confirming that RNase E has a broad impact on the M. tuberculosis transcriptome.

Contaminant Organism Growth in Febrile Infants at Low Risk for Invasive Bacterial Infection.

In this multicenter, cross-sectional, secondary analysis of 4042 low-risk febrile infants, nearly 10% had a contaminated culture obtained during their evaluation (4.9% of blood cultures, 5.0% of urine cultures, and 1.8% of cerebrospinal fluid cultures). Our findings have important implications for improving sterile technique and reducing unnecessary cultures.

CarboGrove: a resource of glycan-binding specificities through analyzed glycan-array datasets from all platforms.

Glycan arrays continue to be the primary resource for determining the glycan-binding specificity of proteins. The volume and diversity of glycan-array data are increasing, but no common method and resource exist to analyze, integrate, and use the available data. To meet this need, we developed a resource of analyzed glycan-array data called CarboGrove. Using the ability to process and interpret data from any type of glycan array, we populated the database with the results from 35 types of glycan arrays, 13 glycan families, 5 experimental methods, and 19 laboratories or companies. In meta-analyses of glycan-binding proteins, we observed glycan-binding specificities that were not uncovered from single sources. In addition, we confirmed the ability to efficiently optimize selections of glycan-binding proteins to be used in experiments for discriminating between closely related motifs. Through descriptive reports and a programmatically accessible Application Programming Interface, CarboGrove yields unprecedented access to the wealth of glycan-array data being produced and powerful capabilities for both experimentalists and bioinformaticians.

Surface-Enhanced Raman Spectroscopy for the Detection of a Metabolic Product in the Headspace Above Live Bacterial Cultures.

In situ surface-enhanced Raman spectra of the headspace above cultures of six bacterial species showed strong characteristic bands from chemisorbed methyl sulfide. This marker compound is created by dissociation of dimethyl disulfide (DMDS), a fermentative metabolite of bacteria, on the surface of the enhancing Au or Ag nanoparticle films. Kinetic binding plots of media spiked with DMDS and of live cultures showed that the Au-based substrates were more suitable for the rapid detection of bacteria than Ag-based substrates. For E. coli DH5α, the sensitivity limit for headspace SERS detection was 1.5×107  CFU mL-1 , which corresponded to detection 15 min after inoculation of the growth medium. Since the metabolites are only produced by viable bacteria, antibiotic (gentamicin) treatment stopped the normal signal growth of the marker peak. This work is a promising step towards rapid bedside detection of bacterial infections and rapid screening of antibiotics against bacteria.

Building pH sensors into paper-based small-molecular logic systems for very simple detection of edges of objects.

Genetically engineered bacteria and reactive DNA networks detect edges of objects, as done in our retinas and as also found within computer vision. We now demonstrate that simple molecular logic systems (a combination of a pH sensor, a photo acid generator, and a pH buffer spread on paper) without any organization can achieve this relatively complex computational goal with good fidelity. This causes a jump in the complexity achievable by molecular logic-based computation and extends its applicability. The molecular species involved in light dose-driven "off-on-off" fluorescence is diverted in the "on" state by proton diffusion from irradiated to unirradiated regions where it escapes a strong quencher, thus visualizing the edge of a mask.

Fluorescent logic systems for sensing and molecular computation: structure-activity relationships in edge-detection.

Molecular logic-based computation continues to throw up new applications in sensing and switching, the newest of which is the edge detection of objects. The scope of this phenomenon is mapped out by the use of structure-activity relationships, where several structures of the molecules and of the objects are examined. The different angles and curvatures of the objects are followed with good fidelity in the visualized edges, even when the objects are in reverse video.

Perceived life threat in children during the COVID-19 pandemic: associations with posttraumatic stress, anxiety, and depressive symptoms.

Defining children's "trauma exposure" in the context of the COVID-19 pandemic has been a source of debate. Children were exposed to threatening messaging about COVID-19 but might interpret this information differently than adults. Perceived life threat (PLT), the belief that one's life is in danger, has been identified as a robust predictor of posttraumatic stress symptoms (PTSS), and may be a better predictor of PTSS than actual life threat (ALT). This study investigated parent reports of children's self-PLT (belief that they might die from COVID-19) and family-PLT (belief that a family member might die from COVID-19). The aims were to compare PLT to ALT, evaluate their associations with children's psychological functioning, and identify risk factors associated with PLT. We hypothesized an association between PLT and children's psychological functioning in the context of the COVID-19 pandemic. Parents (N = 140) reported on their child's (M age = 9.81 years, 47% female) pandemic experiences, psychological functioning, and both self-PLT and family-PLT. Results revealed self-PLT for 10% of the children and family-PLT for 43% of the children, yet only 6% experienced ALT (i.e., they or their parent tested positive for COVID-19). Children with reported self- or family-PLT had higher PTSS, depressive symptoms, anxiety symptoms, and functional impairment compared to children without these reported beliefs. PLT, but not ALT, was associated with psychological outcomes. Children with only PLT had greater PTSS and impairment than children with ALT. There were differences in parental functioning and pandemic-related information/media exposure between children with and without PLT. Children's perceptions, rather than objective experiences, may be more central to their psychological functioning. This has implications for screening for pandemic-related symptomatology in children as traditional trauma exposure measures may not adequately identify distressed children.

Race, Ethnicity, Language, and the Treatment of Low-Risk Febrile Infants.

Importance: Febrile infants at low risk of invasive bacterial infections are unlikely to benefit from lumbar puncture, antibiotics, or hospitalization, yet these are commonly performed. It is not known if there are differences in management by race, ethnicity, or language. Objective: To investigate associations between race, ethnicity, and language and additional interventions (lumbar puncture, empirical antibiotics, and hospitalization) in well-appearing febrile infants at low risk of invasive bacterial infection. Design, Setting, and Participants: This was a multicenter retrospective cross-sectional analysis of infants receiving emergency department care between January 1, 2018, and December 31, 2019. Data were analyzed from December 2022 to July 2023. Pediatric emergency departments were determined through the Pediatric Emergency Medicine Collaborative Research Committee. Well-appearing febrile infants aged 29 to 60 days at low risk of invasive bacterial infection based on blood and urine testing were included. Data were available for 9847 infants, and 4042 were included following exclusions for ill appearance, medical history, and diagnosis of a focal infectious source. Exposures: Infant race and ethnicity (non-Hispanic Black, Hispanic, non-Hispanic White, and other race or ethnicity) and language used for medical care (English and language other than English). Main Outcomes and Measures: The primary outcome was receipt of at least 1 of lumbar puncture, empirical antibiotics, or hospitalization. We performed bivariate and multivariable logistic regression with sum contrasts for comparisons. Individual components were assessed as secondary outcomes. Results: Across 34 sites, 4042 infants (median [IQR] age, 45 [38-53] days; 1561 [44.4% of the 3516 without missing sex] female; 612 [15.1%] non-Hispanic Black, 1054 [26.1%] Hispanic, 1741 [43.1%] non-Hispanic White, and 352 [9.1%] other race or ethnicity; 3555 [88.0%] English and 463 [12.0%] language other than English) met inclusion criteria. The primary outcome occurred in 969 infants (24%). Race and ethnicity were not associated with the primary composite outcome. Compared to the grand mean, infants of families that use a language other than English had higher odds of the primary outcome (adjusted odds ratio [aOR]; 1.16; 95% CI, 1.01-1.33). In secondary analyses, Hispanic infants, compared to the grand mean, had lower odds of hospital admission (aOR, 0.76; 95% CI, 0.63-0.93). Compared to the grand mean, infants of families that use a language other than English had higher odds of hospital admission (aOR, 1.08; 95% CI, 1.08-1.46). Conclusions and Relevance: Among low-risk febrile infants, language used for medical care was associated with the use of at least 1 nonindicated intervention, but race and ethnicity were not. Secondary analyses highlight the complex intersectionality of race, ethnicity, language, and health inequity. As inequitable care may be influenced by communication barriers, new guidelines that emphasize patient-centered communication may create disparities if not implemented with specific attention to equity.

Changes in Pediatricians' Child Passenger Safety Counseling With Initiation of a Booster Seat Program.

Booster seats reduce injury in motor vehicle crashes, yet they are used less frequently than car seats and seat belts. Primary care providers are well positioned to educate and encourage families to use booster seats. We aimed to assess how a booster seat distribution program affected the documentation of restraint usage and anticipatory guidance at well-child visits at a pediatric primary care practice. We performed a retrospective chart review of patients aged 4 to 12 years from June to December in 2019 and 2020, representing before and after a booster seat program. The most frequently documented restraints in 2019 and 2020 were seat belts (51% vs 30%), booster seats (25% vs 27%), and not documented/unclear (17% vs 25%) (P < .001). The program significantly increased referrals for booster seats (P < .001). Despite significant differences in the proportion of children in each restraint category, overall booster seat use was similar between years.

Continuing Education Evaluations of an Internal Medicine and a Pediatric Podcast.

INTRODUCTION: This study aimed to characterize practitioners who use podcasts as a form of continuing education (CE), evaluate attitudes regarding podcasting as a medium for CE, and assess intended practice change after listening to podcasts for CE. METHODS: We examined CE data from a mandatory postpodcast evaluation collected between February 2021 and August 2021 for two free podcasts. We analyzed linked episode data containing podcast downloads. RESULTS: Listeners downloaded 972,691 episodes and claimed 8182 CE credits (less than 1% of total downloads) over 7 months. CE credit was claimed by physicians, physician assistants, nurses, and pharmacists. Most listeners claiming CE credit were not affiliated with an academic institution. Motivation for listening to episodes included a topic of interest, relevance to a patient, and a topic of less comfort. Of individuals obtaining CE, 98% intended to implement change after listening. DISCUSSION: Although only a small number of podcast listeners claim CE credit, those who claim credit represent a diverse and interprofessional audience. Listeners select podcasts to fill self-identified learning needs. Overwhelmingly, listeners report podcast CE changes intended practice. Podcasts may be an effective modality for CE and practice change; further research should focus on facilitators and barriers to CE uptake and on patient health outcomes.

Mammalian sleep genetics.

Mammalian sleep is a complex phenomenon governed by the interplay of neural circuits and signaling systems. The impact of genetic manipulations on sleep-wake dynamics provides important insights into this complex behavior. Here we review the sleep-related phenotypes of over 50 transgenic animal models spanning a variety of signaling systems. This heterogeneous literature includes outcomes spanning motor activity patterns, sleep-wake stage architecture, responses to sleep deprivation, circadian rhythmicity, and other perturbations such as food restriction, temperature challenge, and infection exposure. Insights from these animal experiments hold potential to converge with the well-known sleep-wake neurocircuitry as well as the increasingly available human genetic information, especially in patient populations exhibiting sleep-wake pathology.

Give Me A Boost: A Child Passenger Safety Educational Intervention.

BACKGROUND: Motor vehicle crashes (MVCs) are a leading cause of morbidity and mortality among children, yet belt-positioning booster seats remain underutilized. This study analyzed the impact of a provider-oriented child passenger safety (CPS) educational intervention on pediatric resident confidence, knowledge, and counseling behavior in the primary care setting. METHODS: Pre- and post-educational intervention cross- sectional surveys were distributed to pediatric residents focusing on confidence, knowledge, barriers to, and frequency of CPS discussions. RESULTS: Pre-intervention, only 16% (95% CI: 6.2-32%) of respondents reported confidence in their knowledge of the American Academy of Pediatrics' (AAP) booster seat recommendations. Post-intervention frequency of CPS discussion, confidence and knowledge in all measured aspects increased. CONCLUSIONS: While pediatric residents are well positioned for CPS counseling, they lack adequate baseline knowledge of CPS recommendations, particularly for booster seats. Brief educational interventions can increase and maintain provider knowledge and confidence in counseling families on appropriate child car safety seats.

Cardiopulmonary Resuscitation Training in Select Rhode Island High Schools: A Descriptive Study.

Fewer than 15% of people who have out-of-hospital cardiac arrests survive, but chances of survival can be tripled with effective bystander cardiopulmonary resuscitation (CPR). The majority of states, including Rhode Island, require high school CPR training, yet the impact of this is not well studied. A 33-question REDCap survey regarding cardiac arrest preparedness, CPR education, and barriers to CPR training was emailed to high school staff in Rhode Island. There were 62 responses; 26% reported their school taught CPR and 94% felt it was important for students to have CPR certification. Barriers included time (85%), budget (82%), and materials (79%). Over 80% felt students would not be able to perform high-quality CPR or properly use a defibrillator. Despite laws requiring CPR training and the belief by school staff of the importance of CPR training, the majority of students are not receiving CPR training. Staff report students do not have the ability to perform effective CPR or use a defibrillator.

Learning Through Listening: A Scoping Review of Podcast Use in Medical Education.

PURPOSE: To improve understanding of podcast use in medical education by examining current research on descriptive attributes and educational outcomes, highlighting implications of the current evidence base for educational practices, and identifying research gaps to guide future investigation. METHOD: The authors conducted a scoping review, searching PubMed and Embase databases in June-July 2020 for English-language studies of audio-only medical education podcast use in undergraduate, graduate, and continuing medical education. The authors excluded studies without original data or with nonphysician data that could not be separated from physician data. From included studies, the authors extracted data regarding descriptive outcomes (e.g., podcast use, content areas, structure) and educational outcomes (classified using Kirkpatrick's 4 levels of evaluation). RESULTS: Of 491 unique articles, 62 met inclusion criteria. Descriptive outcomes were reported in 44 studies. Analysis of these studies revealed podcast use has increased over time, podcasts are a top resource for resident education, and podcasts are being incorporated into formal medical curricula. Educational outcomes were reported in 38 studies. The 29 studies that assessed learner reaction and attitudes to podcasts (Kirkpatrick level 1) showed learners value podcasts for their portability, efficiency, and combined educational and entertainment value. The 10 studies that assessed knowledge retention (Kirkpatrick level 2) showed podcasts to be noninferior to traditional teaching methods. The 11 studies that assessed behavior change (Kirkpatrick level 3) showed improved documentation skills in medical students and self-reported practice change in residents and practicing physicians after listening to podcasts. None of the studies reported system change or patient outcomes (Kirkpatrick level 4). CONCLUSIONS: Future research should focus on the optimal structure of podcasts for learning, higher-level outcomes of podcasts, and the implementation of podcasts into formal curricula. Podcasts may prove to be essential tools for disseminating and implementing the most current, evidence-based practices.

Atopy and Lifestyle Survey of Allergic Patients From Urban Environment in Romania: Preliminary Data From an Interactive Qualifying Project.

Introduction Respiratory allergies represent an important public health problem, with increasing prevalence and severity in Europe during the last decades. The rise of pollen allergies is an issue that continues to negatively impact people's daily lives across the globe and has become more important in the light of global warming and increasing air pollution. The aim of our paper is to evaluate the prevalence of declared atopy and the influence of lifestyle on allergic diseases, particularly on pollen allergies, in the urban environment from Romania. Methods The study is based on the cooperation Interactive Qualifying Project (IQP) called "Pollen Allergies in Romania: Optimizing Data Analysis in Raising Awareness", agreed and carried on between a group of North-American students and teachers from Worcester Polytechnic Institute and a hospital-based allergy team from Carol Davila University of Medicine and Pharmacy and Colentina Clinical Hospital from Bucharest. The project aimed to evaluate the prevalence of atopy and lifestyle practices of allergic patients and to develop a data analysis tool to determine correlations between pollen counts and other environmental factors in the city of Bucharest. Results The lifestyle survey revealed that about one-third of allergic patients declared history of atopy. Some of the declared lifestyle practices can be considered environmental risk factors for allergies. This IQP can be considered a model of international, interdisciplinary and intercultural collaboration. Conclusion We concluded that Romania is facing an increasing pollen allergies trend and some actual lifestyle aspects can significantly influence the risk of pollen allergies in the big city environment.

Changes in Sick Visits at an Academic Pediatric Primary Care Practice due to the COVID-19 Pandemic.

BACKGROUND/OBJECTIVE: The COVID-19 pandemic decreased pediatric patient volumes; however, details regarding patterns of use within primary care sick visits are not well understood. METHODS: We performed a retrospective chart review of sick visits in an academic primary care clinic from March-August 2019 and 2020 and recorded demographics and visit diagnoses. Descriptive statistics, Chi-square, and Fisher's exact tests were used to compare the two time periods. RESULTS: Patient age, gender, and insurance type were similar across years. In 2020, there were 1,868 sick visits (247 telehealth, 4%-36% of monthly visits) compared to 4,007 (0 telehealth) in 2019. The proportion of infectious diagnoses decreased (35% vs 48%); non-infectious diagnoses increased, including dermatological (25% vs 19%) and genitourinary/reproductive (9% vs. 6%) diagnoses.  Conclusion: Similar to pediatric emergency departments, we found decreased primary care sick visits. Telehealth increased in 2020 and varied with COVID-19 community prevalence. Visits for contagious illnesses decreased in 2020, likely related to mitigation measures.

Visits for Possible COVID-19 in a Pediatric Primary Care Practice Early in the Pandemic.

BACKGROUND: Children with COVID-19 usually present with mild symptoms. We characterize visits with respect to symptoms and testing in the outpatient setting. METHODS: A retrospective chart review of sick visits in a pediatric academic primary care clinic April-August 2020. We included possible COVID-19 cases, or "persons under investigation" (PUIs), recording symptoms, positive contacts, and COVID-19 testing. Descriptive statistics and Chi-square or Fisher's exact tests for comparisons were used. RESULTS: 32% (476/1,474) of sick visits were PUIs; 20% were telehealth. Symptoms most commonly reported were fever, congestion/rhinorrhea and cough. 76% of PUIs were tested for COVID-19. Only presence of COVID-19 contacts and loss of taste/smell were significantly associated with positive tests (p<0.001). CONCLUSION: Nearly a third of sick visits in an academic pediatric practice were seen for possible COVID-19 symptoms and most were tested. The majority with and without COVID-19 had fever, congestion and/or cough. Our findings suggest low thresholds for testing in children.

Neck circumference in Latin America and the Caribbean: A systematic review and meta-analysis.

Background: High neck circumference (NC) is associated with high burden diseases in Latin American and the Caribbean (LAC). NC complements established anthropometric measurements for early identification of cardio-metabolic and other illnesses. However, evidence about NC has not been systematically studied in LAC. We aimed to estimate the mean NC and the prevalence of high NC in LAC. Methods: We conducted a systematic review in MEDLINE, Embase, Global Health and LILACS. Search results were screened and studied by two reviewers independently. To assess risk of bias of individual studies, we used the Hoy et al. scale and the Newcastle-Ottawa scale. We conducted a random-effects meta-analysis. Results: In total, 182 abstracts were screened, 96 manuscripts were reviewed and 85 studies (n= 51,978) were summarized. From all the summarized studies, 14 were conducted in a sample of the general population, 23 were conducted with captive populations and 49 studies were conducted with patients. The pooled mean NC in the general population was 35.69 cm (95% IC: 34.85cm-36.53cm; I²: 99.6%). In our patient populations, the pooled mean NC in the obesity group was 42.56cm (95% CI 41.70cm-43.42cm; I²: 92.40%). Across all studied populations, there were several definitions of high NC; thus, prevalence estimates were not comparable. The prevalence of high NC ranged between 37.00% and 57.69% in the general population. The methodology to measure NC was not consistently reported. Conclusions: Mean NC in LAC appears to be in the range of estimates from other world regions. Inconsistent methods and definitions hamper cross-country comparisons and time trend analyses. There is a need for consistent and comparable definitions of NC so that it can be incorporated as a standard anthropometric indicator in surveys and epidemiological studies.