Eos Promotes TH2 Differentiation by Interacting with and Propagating the Activity of STAT5.

The Ikaros zinc-finger transcription factor Eos has largely been associated with sustaining the immunosuppressive functions of regulatory T cells. Paradoxically, Eos has more recently been implicated in promoting proinflammatory responses in the dysregulated setting of autoimmunity. However, the precise role of Eos in regulating the differentiation and function of effector CD4+ T cell subsets remains unclear. In this study, we find that Eos is a positive regulator of the differentiation of murine CD4+ TH2 cells, an effector population that has been implicated in both immunity against helminthic parasites and the induction of allergic asthma. Using murine in vitro TH2 polarization and an in vivo house dust mite asthma model, we find that EosKO T cells exhibit reduced expression of key TH2 transcription factors, effector cytokines, and cytokine receptors. Mechanistically, we find that the IL-2/STAT5 axis and its downstream TH2 gene targets are one of the most significantly downregulated pathways in Eos-deficient cells. Consistent with these observations, we find that Eos forms, to our knowledge, a novel complex with and supports the tyrosine phosphorylation of STAT5. Collectively, these data define a regulatory mechanism whereby Eos propagates STAT5 activity to facilitate TH2 cell differentiation.

Cultural and Linguistic Adaptations of Stop the Bleed in Multi-Ethnic Refugee Communities.

INTRODUCTION: Stop the Bleed (STB), and other trainings that promote health education in basic trauma management techniques, is offered mostly in English and Spanish in the United States. Limited access to injury prevention training may contribute to inequities in health outcomes for individuals with limited English proficiency (LEP). Our study aims to determine the feasibility and effectiveness of STB training in 4 languages spoken in a super diverse refugee settlement community, Clarkston, GA. METHODS: Written STB educational materials were culturally adapted, translated, and back translated into 4 languages: Arabic, Burmese, Somali, and Swahili. Four 90-minute in-person STB trainings were conducted by medical personnel with community-based interpreters at a central and familiar location in the Clarkston community. Pre- and post-tests were administered in participant's preferred language to evaluate change in knowledge and beliefs as well as the effectiveness of the training method. RESULTS: A total of 46 community members were trained in STB, the majority of which were women (63%). Participants demonstrated improvement in their knowledge, confidence, and comfort using STB techniques. Participants reported that 2 aspects of the training were particularly beneficial: the presence of language concordant interpreters from the community and small group hands on sessions that allowed for practicing STB techniques. CONCLUSION: Cultural and linguistic adaptation of STB training is a feasible, cost-effective, and effective method for disseminating life-saving information and trauma education to immigrant populations who have LEP. Expansion of community training and partnerships to support the needs of diverse communities is both necessary and urgent.

The Effect COVID Has Had on the Wants and Needs of Children in Terms of Play: Text Mining the Qualitative Response of the Happen Primary School Survey with 20,000 Children in Wales, UK between 2016 and 2021.

Play is central to children's physical and social development. This study examines changes in children's response to questions on play opportunities between 2016 and 2021. Primary school children aged 8-11 in Wales participated in the HAPPEN survey between 2016 and 2021. The survey captures a range of information about children's health and wellbeing, including open-ended questions about what could make them happier. Text mining methods were used to examine how open-ended responses have changed over time in relation to play, before and, after the COVID enforced school closures. A total of 20,488 participant responses were analysed, 14,200 pre-school closures (2016 to pre-March 2020) and 6248 after initial school closures (September 2020-December 2021). Five themes were identified based on children's open-ended responses; (a) space to play (35%), (b) their recommendations on play (31%), (c) having permission to play (20%), (d) their feelings on health and wellbeing and play (10%) and (e) having time to play (4%). Despite differences due to mitigation measures, the predominant recommendation from children after COVID is that they would like more space to play (outside homes, including gardens), more time with friends and protected time to play with friends in school and at home.

Systematic content analysis of patient evaluations of START NOW psychotherapy reveals practical strategies for improving the treatment of opioid use disorder.

BACKGROUND: Clinical trials provide consistent evidence for buprenorphine's efficacy in treating opioid use disorder (OUD). While the Drug Addiction Treatment Act of 2000 requires physicians to combine medication-assisted treatment (MAT) with behavioral intervention, there is no clear evidence for what form or elements of psychotherapy are most effective when coupled with MAT to treat OUD. This investigation involves focus groups designed to collect patient opinions about a specific psychotherapy, called START NOW, as well as general beliefs about various elements of psychotherapy for treating OUD. Our analysis reveals trends about patient preferences and strategies for improving OUD treatment. METHODS: Subjects included patients enrolled in buprenorphine/naloxone MAT at our institution's office-based opioid treatment program. All subjects participated in a single START NOW group session, which was led by a provider (physician or nurse practitioner trained and standardized in delivering START NOW). Consented subjects participated in satisfaction surveys and audio-recorded focus groups assessing individual beliefs about various elements of psychotherapy for treating OUD. RESULTS: Overall, 38 different focus groups, 92 participation events, and 44 unique subjects participated in 1-to-6 different START NOW session/audio-recorded focus group sessions led by a certified moderator. Demographic data from 36/44 subjects was collected. Seventy-five percent (33/44) completed the START NOW Assessment Protocol, which revealed self-reported behavioral trends. Analysis of all 92 START NOW Satisfaction Questionnaire results suggests that subjects' opinions about START NOW improved with increased participation. Our analysis of audio-recorded focus groups is divided into three subsections: content strategies for new psychotherapies, implementation strategies, and other observations. For example, participants request psychotherapies to target impulsivity and to teach future planning and build positive relationships. CONCLUSIONS: The results of this study may guide implementation of psychotherapy and improve the treatment of OUD, especially as it relates to improving the modified START NOW program for treating OUD. Our study also reveals a favorable outlook of START NOW with increased participation, suggesting that any initial reticence to this program can be overcome to allow for effective implementation.

Peritonsillar Abscess Model for Ultrasound Diagnosis Using Inexpensive Materials.

Audience: This low-cost peritonsillar abscess model is intended for the education of emergency medicine and otolaryngology residents and advanced care practitioners of all training levels. Introduction: With incidence rates as high as 124 per 100,000 in the 14-21 age range, peritonsillar abscesses (PTA) are one of the more common head and neck soft tissue infections encountered in the emergency department.1 Peritonsillar abscesses can present to the emergency department in critically ill patients with the dangers of airway compromise and further local spreading. Emergency medicine (EM) residents need practice to properly identify and to minimize procedural complications such as perforation of nearby vessels, aspiration pneumonitis, and airway compromise. A major tool used in the emergency department that can help prevent complications is the use of ultrasound, which the Accreditation Council for Graduate Medical Education (ACGME) requires residents to become proficient at.2 Historically, computed tomography (CT) scanning to diagnose along with blind drainage has been the method of choice. With a sensitivity of 95.2%, intraoral ultrasound can minimize both radiation and procedure related complications.3 The current simulators available come at significant capital expenditure and do not provide high-fidelity ultrasound experience. Here we design and implement a low-cost trainer for residents to use ultrasound to diagnose and drain a PTA. Educational Objectives: By the end of this instructional session learners should be able to: 1) identify and discuss the indications, contraindications, and complications associated with peritonsillar abscesses, 2) properly identify and measure a PTA through ultrasound, and 3) competently perform ultrasound-guided peritonsillar abscess drainage on a simulator and remove fluid. Educational Methods: This PTA model utilizes task trainers designed from Styrofoam wig heads. An airway was modeled using readily available wood shop tools and balloons filled with a fluid mixture containing coconut lotion, water, and fragrance beads, which were inserted into the airway. This unique mixture within the balloons creates a realistic echogenicity of an abscess with loculations. With emergency medicine clinical faculty guidance and the use of ultrasound, learners are able to identify a peritonsillar abscess and subsequently demonstrate drainage of fluid with a needle and syringe. Research Methods: This PTA model was tested with a group of 36 emergency medicine residents. Optional, anonymous post surveys were completed by 17 residents. A 5-point Likert Scale was used to assess utility of this model. Results: The majority of users agreed the model provides a realistic image of the disease for diagnosis by ultrasound with a score of 3.6 and felt more comfortable identifying and draining peritonsillar abscesses with scores of 3.7 and 3.6 respectively. Learners' surveys revealed the session was useful and improved their knowledge with both scoring 3.8. No critical feedback was given by learners or instructors. The efficacy of the content was assessed by evaluators observing proper ultrasound, procedure set up, and drainage of PTA. Discussion: This inexpensive model to expose residents to proper PTA drainage was effective considering learners' high response to post-procedure survey scales. The results of our pilot implementation showed this model has utility in teaching ultrasound guided identification and drainage of PTA's. With minimal build and optimized instruction time, we can improve residents' comfort in performing this procedure and allow for important simulation experience in a safe, controlled environment. Topics: Simulation, emergency medicine, peritonsillar abscess, otolaryngology.

Antibody responses to Zika virus proteins in pregnant and non-pregnant macaques.

The specificity of the antibody response against Zika virus (ZIKV) is not well-characterized. This is due, in part, to the antigenic similarity between ZIKV and closely related dengue virus (DENV) serotypes. Since these and other similar viruses co-circulate, are spread by the same mosquito species, and can cause similar acute clinical syndromes, it is difficult to disentangle ZIKV-specific antibody responses from responses to closely-related arboviruses in humans. Here we use high-density peptide microarrays to profile anti-ZIKV antibody reactivity in pregnant and non-pregnant macaque monkeys with known exposure histories and compare these results to reactivity following DENV infection. We also compare cross-reactive binding of ZIKV-immune sera to the full proteomes of 28 arboviruses. We independently confirm a purported ZIKV-specific IgG antibody response targeting ZIKV nonstructural protein 2B (NS2B) that was recently reported in ZIKV-infected people and we show that antibody reactivity in pregnant animals can be detected as late as 127 days post-infection (dpi). However, we also show that these responses wane over time, sometimes rapidly, and in one case the response was elicited following DENV infection in a previously ZIKV-exposed animal. These results suggest epidemiologic studies assessing seroprevalence of ZIKV immunity using linear epitope-based strategies will remain challenging to interpret due to susceptibility to false positive results. However, the method used here demonstrates the potential for rapid profiling of proteome-wide antibody responses to a myriad of neglected diseases simultaneously and may be especially useful for distinguishing antibody reactivity among closely related pathogens.

Highly efficient maternal-fetal Zika virus transmission in pregnant rhesus macaques.

Infection with Zika virus (ZIKV) is associated with human congenital fetal anomalies. To model fetal outcomes in nonhuman primates, we administered Asian-lineage ZIKV subcutaneously to four pregnant rhesus macaques. While non-pregnant animals in a previous study contemporary with the current report clear viremia within 10-12 days, maternal viremia was prolonged in 3 of 4 pregnancies. Fetal head growth velocity in the last month of gestation determined by ultrasound assessment of head circumference was decreased in comparison with biparietal diameter and femur length within each fetus, both within normal range. ZIKV RNA was detected in tissues from all four fetuses at term cesarean section. In all pregnancies, neutrophilic infiltration was present at the maternal-fetal interface (decidua, placenta, fetal membranes), in various fetal tissues, and in fetal retina, choroid, and optic nerve (first trimester infection only). Consistent vertical transmission in this primate model may provide a platform to assess risk factors and test therapeutic interventions for interruption of fetal infection. The results may also suggest that maternal-fetal ZIKV transmission in human pregnancy may be more frequent than currently appreciated.

Heterologous Protection against Asian Zika Virus Challenge in Rhesus Macaques.

BACKGROUND: Zika virus (ZIKV; Flaviviridae, Flavivirus) was declared a public health emergency of international concern by the World Health Organization (WHO) in February 2016, because of the evidence linking infection with ZIKV to neurological complications, such as Guillain-Barre Syndrome in adults and congenital birth defects including microcephaly in the developing fetus. Because development of a ZIKV vaccine is a top research priority and because the genetic and antigenic variability of many RNA viruses limits the effectiveness of vaccines, assessing whether immunity elicited against one ZIKV strain is sufficient to confer broad protection against all ZIKV strains is critical. Recently, in vitro studies demonstrated that ZIKV likely circulates as a single serotype. Here, we demonstrate that immunity elicited by African lineage ZIKV protects rhesus macaques against subsequent infection with Asian lineage ZIKV. METHODOLOGY/PRINCIPAL FINDINGS: Using our recently developed rhesus macaque model of ZIKV infection, we report that the prototypical ZIKV strain MR766 productively infects macaques, and that immunity elicited by MR766 protects macaques against heterologous Asian ZIKV. Furthermore, using next generation deep sequencing, we found in vivo restoration of a putative N-linked glycosylation site upon replication in macaques that is absent in numerous MR766 strains that are widely being used by the research community. This reversion highlights the importance of carefully examining the sequence composition of all viral stocks as well as understanding how passage history may alter a virus from its original form. CONCLUSIONS/SIGNIFICANCE: An effective ZIKV vaccine is needed to prevent infection-associated fetal abnormalities. Macaques whose immune responses were primed by infection with East African ZIKV were completely protected from detectable viremia when subsequently rechallenged with heterologous Asian ZIKV. Therefore, these data suggest that immunogen selection is unlikely to adversely affect the breadth of vaccine protection, i.e., any Asian ZIKV immunogen that protects against homologous challenge will likely confer protection against all other Asian ZIKV strains.

A rhesus macaque model of Asian-lineage Zika virus infection.

Infection with Asian-lineage Zika virus (ZIKV) has been associated with Guillain-Barré syndrome and fetal abnormalities, but the underlying mechanisms remain poorly understood. Animal models of infection are thus urgently needed. Here we show that rhesus macaques are susceptible to infection by an Asian-lineage ZIKV closely related to strains currently circulating in the Americas. Following subcutaneous inoculation, ZIKV RNA is detected in plasma 1 day post infection (d.p.i.) in all animals (N=8, including 2 pregnant animals), and is also present in saliva, urine and cerebrospinal fluid. Non-pregnant and pregnant animals remain viremic for 21 days and for up to at least 57 days, respectively. Neutralizing antibodies are detected by 21 d.p.i. Rechallenge 10 weeks after the initial challenge results in no detectable virus replication, indicating protective immunity against homologous strains. Therefore, Asian-lineage ZIKV infection of rhesus macaques provides a relevant animal model for studying pathogenesis and evaluating potential interventions against human infection, including during pregnancy.