Deep mutational scanning reveals a correlation between degradation and toxicity of thousands of aspartoacylase variants.

Unstable proteins are prone to form non-native interactions with other proteins and thereby may become toxic. To mitigate this, destabilized proteins are targeted by the protein quality control network. Here we present systematic studies of the cytosolic aspartoacylase, ASPA, where variants are linked to Canavan disease, a lethal neurological disorder. We determine the abundance of 6152 of the 6260 ( ~ 98%) possible single amino acid substitutions and nonsense ASPA variants in human cells. Most low abundance variants are degraded through the ubiquitin-proteasome pathway and become toxic upon prolonged expression. The data correlates with predicted changes in thermodynamic stability, evolutionary conservation, and separate disease-linked variants from benign variants. Mapping of degradation signals (degrons) shows that these are often buried and the C-terminal region functions as a degron. The data can be used to interpret Canavan disease variants and provide insight into the relationship between protein stability, degradation and cell fitness.

Long-acting reversible contraception use and unmet desire among patients after the Zika Contraception Access Network Program in Puerto Rico.

OBJECTIVES: To describe unmet desire for long-acting reversible contraception (LARC) after the Zika Contraception Access Network (Z-CAN) in Puerto Rico during the 2016-2017 Zika outbreak. STUDY DESIGN: Z-CAN patients completed surveys about contraception experiences over a 3-year period. RESULTS: Of 1809 respondents, 3% never used LARC but reported wanting it since their initial visit. As reasons for not getting LARC, nearly 50% indicated a provider-related reason and 25% reported cost. CONCLUSIONS: Few Z-CAN patients who never used LARC had unmet desire. Provider training in contraception guidelines and strategies to address costs can expand access to the full range of reversible contraception. IMPLICATIONS: Three years after a short-term program provided reversible contraception in Puerto Rico, few respondents had never used but wanted a long-acting reversible contraception method. Nearly half reported provider-related reasons for not receiving long-acting reversible contraception, and 25% reported cost. Provider awareness of contraceptive guidance and method availability can support client-centered care.

Perceptions of the Zika Virus, Contraceptive Access, and Motivation to Participate in the Zika Contraception Access Network Program: Qualitative Analysis of Focusgroup Discussions with Puerto Rican Women.

OBJECTIVE: During the 2016-2017 Zika virus outbreak in Puerto Rico, the Zika Contraception Access Network (Z-CAN) provided client-centered contraceptive counseling and access to the full range of reversible contraceptive methods at no cost to prevent unintended pregnancies and thereby to reduce Zika-related birth outcomes. METHODS: To understand how Puerto Rican women's perceptions of the Zika virus affected contraceptive decisions and assess how they heard about the Z-CAN program and what influenced their participation, or lack thereof, 24 focus-group discussions were conducted among women of reproductive age who did and did not participate in Z-CAN. RESULTS: Women who participated in the discussions often had heard about Z-CAN from their physician or friends; non-participants had heard about Z-CAN from Facebook or friends. Women expressed satisfaction on finding a Z-CAN clinic and valued the same-day provision of contraceptives. When a preferred contraceptive method or a first appointment was not readily available, women reconsidered accessing the program. Women's perceptions and trust of reproductive healthcare providers, their engagement in social networks, and their ability to choose a contraceptive method that best meets their needs can influence participation in contraception-access programs. CONCLUSION: Focus groups can be used to understand women's knowledge of the Zika virus, barriers and facilitators to contraception access, and motivations for participation in the Z-CAN program.

Community-based organizations at the front lines of the COVID-19 pandemic: Strategies for building equitable COVID-19 vaccine uptake and distribution.

The coronavirus disease 2019 (COVID-19) pandemic highlighted the disproportionate negative health outcomes faced by racial and ethnic minority communities as a result of various factors that affect health equity. In response to the COVID-19 pandemic and the overwhelming disparities within communities, public health agencies have aimed to establish partnerships with community-based organizations (CBOs) to distribute information about COVID-19 to populations expressing vaccine hesitancy, recognizing that they are necessary partners in ensuring and maintaining equitable distribution of resources. In line with the National Foundation for the Centers for Disease Control and Prevention (CDC Foundation)'s Health Equity Strategy, the CDC Foundation's Response, Crisis and Preparedness Unit (RCPU) team is responsible for managing the portfolio of more than 110 grants to CBOs across 14 grant opportunities by private and federal funding that supports the COVID-19 response. The goal is to promote activities, including support for sharing of accurate, culturally appropriate vaccine information to address concerns and the development and strengthening of strategic partnerships to support effective community outreach and vaccination. Most, if not all of the CBOs use similar techniques to address equitable vaccine uptake and distribution by partnering or becoming vaccine providers via mobile or pop-up clinics, engaging community health workers, door-to-door canvasing, conducting listening sessions, providing incentives, and hosting community events and communications campaigns. As of June 23, 2022, the funded CBOs have collectively reached over 14.8 million individuals with COVID-19 safety and vaccine education messaging, administered 288,197 COVID-19 vaccinations within their communities, and partnered with 416 public health jurisdictions and 853 community organizations. Two case studies are provided to share specifics on strategies and activities done toward providing equitable information and vaccine distribution. This rapid and flexible grant management process al-lowed CBOs to focus on the areas of greatest need in their community and select strategies that would be well-received by community members.

A mutational atlas for Parkin proteostasis.

Proteostasis can be disturbed by mutations affecting folding and stability of the encoded protein. An example is the ubiquitin ligase Parkin, where gene variants result in autosomal recessive Parkinsonism. To uncover the pathological mechanism and provide comprehensive genotype-phenotype information, variant abundance by massively parallel sequencing (VAMP-seq) is leveraged to quantify the abundance of Parkin variants in cultured human cells. The resulting mutational map, covering 9219 out of the 9300 possible single-site amino acid substitutions and nonsense Parkin variants, shows that most low abundance variants are proteasome targets and are located within the structured domains of the protein. Half of the known disease-linked variants are found at low abundance. Systematic mapping of degradation signals (degrons) reveals an exposed degron region proximal to the so-called "activation element". This work provides examples of how missense variants may cause degradation either via destabilization of the native protein, or by introducing local signals for degradation.

The Stress of Advancement: A Nurse Practitioner's Exploration in Providing Culturally Competent Obesity Prevention Counseling in Black Women.

BACKGROUND: Black women are diagnosed, disabled, and die from obesity and associated chronic diseases at higher rates than any other sex or race. Advanced practice registered nurses (APRN) can potentially improve culturally relevant health education and counseling by using health literacy communication tools. OBJECTIVE: Explore individualized barriers and APRNs' role in providing obesity prevention education and counseling by assessing the efficacy of the Teach-Back Method (TBM) to understand health habits and attitudes. METHODS: Black women aged 18-45, previously diagnosed as overweight or obese, and identified with perceived barriers were recruited from a predominantly Black church in Atlanta. They engaged in weekly, 1-hour educational sessions via Zoom, addressing four common barriers identified in the literature. Sessions ended with a 5-10 minute Teach-Back session. Pre- and post-intervention Readiness to Change Questionnaire (RCQ) were completed. Descriptive statistics and quantitative data from surveys and pre- and post-RCQ were analyzed. RESULTS: Twenty women completed the intervention. Paired sample t-test revealed no statistical significance or correlation between pre- and post-RCQ scores after using TBM in educational sessions. However, Pearson's correlation showed positive associations between elevated body mass index levels as one advances their education and annual income, with a p-value of 0.05. DISCUSSION: Increased rates of obesity are experienced despite higher educational attainment or pay. Stress and high-coping mechanisms contributed to disordered eating, decreased physical activity engagement, and decreased motivation toward habit change. Clinicians should be held accountable for delivering culturally sensitive care using the TBM, addressing social determinants of health, performing routine stress assessments, and checking their implicit biases.

Maximizing Student Outcomes in Schools: Data-Driven Individualized Education Program Goals and Objectives Aligned to the Standards.

PURPOSE: Federal and state laws require schools to be accountable for student performance on measures of academic achievement in literacy, mathematics, and science skills; monitor high school graduation rates; and track student growth and academic progression throughout the grade levels. Success on these measures gives students pathways to postsecondary options in the workforce, technical education, or college/university education. Speech-language pathologists (SLPs) can utilize existing data sources in the school in conjunction with their knowledge of diagnostics and treatment of cognition, language, and culture to maximize student outcomes beyond the therapy room and in the curriculum standards. METHOD: This clinical focus article will review methods of data collection from existing sources in the students' grade level and academic content curriculum. Utilization of information and input from parents, teachers, paraprofessionals, and others who support the student's academic endeavors will be discussed for Individualized Education Program Present Levels of Academic Achievement and Functional Performance and goal development. Procedures for the SLP to collect independent data aligned to standards-based curriculum will be explored. Consideration will be given for utilizing data to develop future goals and objectives. The benefits of data collection for monitoring student progress and success in the grade-level or alternate standards will be highlighted through case examples. Techniques for classroom-based services, model lessons, coaching, and co-teaching will be presented. CONCLUSIONS: SLPs are recognized as specialized instructional support personnel who have unique knowledge and expertise for diagnosing and treating speech/language impairments (S/LI) disabilities. By understanding how they can use existing data and apply it to developing intervention plans, SLPs can ensure students with S/LI disabilities experience the maximum potential for outcomes in academic and functional success. Data-driven tools and methods aligned to the grade-level standards and curriculum are one of the most powerful tools in an SLP's toolbox. SUPPLEMENTAL MATERIAL: https://doi.org/10.23641/asha.24869592.

Importance of Public and Private Partnership Supporting Data Disaggregation to Measure Racial, Sexual Orientation, and Gender Identity Disparities in COVID-19.

The COVID-19 pandemic has exposed shortcomings in the US public health data system infrastructure, including incomplete or disparate processes related to data collection, management, sharing, and analysis. Public health data modernization is critical to ensure health equity is at the core of preparedness and response efforts and policies that prioritize equitable responses to health emergencies. To address the inequitable uptake and distribution of COVID-19 vaccinations in communities most disproportionately impacted by the pandemic, the CDC Foundation's Response Crisis and Preparedness Unit began partnering with community-based organizations in March 2021 to provide education and outreach and facilitate access to vaccines. These organizations engaged with partners and communities to address vaccine-related concerns, develop innovative and culturally appropriate communication strategies, and promote timely vaccination. Two grantees, Out Boulder County in Colorado and the Coalition of Asian American Leaders in Minnesota, experienced issues related to public health data collection standards and practices for COVID-19. Data collection tools often lack the appropriate or necessary demographic variables or level of disaggregation needed to be able to assess prioritization and disparities within racial and ethnic groups and across sexual orientation and gender identity categories. In this case study, both grantee organizations document their experiences, challenges, and strategies to overcome barriers to implementing their projects resulting from a lack of meaningful data. These examples identify inequities and systems-level changes related to data collection and surveillance, and they provide recommendations and lessons learned to improve data surveillance for more equitable public health responses.

Provider Perceptions of Facilitators of and Barriers to Implementation of the Zika Contraception Access Network: A Qualitative Evaluation.

OBJECTIVE: From May 2016 through September 2017, the Zika Contraception Access Network (Z-CAN) program increased access to contraception during the Zika virus outbreak in Puerto Rico by providing no-cost client-centered contraceptive counseling and (same-day) access to the full range of US Food and Drug Administration-approved reversible contraceptives to women desirous of not becoming pregnant. The purpose of this study was to identify areas for programmatic improvement and enhance the sustainability of services from the perspectives of participating Z-CAN physicians and other staff. METHODS: From April through July 2017, 49 in-depth key-informant interviews were conducted with Z-CAN physicians and clinic staff. Twenty-five clinics participating in the Z-CAN program were selected through a cluster randomization process. A semi-structured interview guide was developed to explore the participants' perceptions of the Z-CAN program and examine facilitators of and barriers to said implementation. A thematic analysis of the emerging topics was conducted. RESULTS: Our analysis encountered 4 common overarching themes: facilitators of the Z-CAN program; barriers to Z-CAN implementation; the perceived impact of Z-CAN on providers and communities; and the sustainability of contraception access after the Z-CAN program ended. The key findings were that provider training, mentor support, and communication campaigns facilitated program implementation and that delays in the acquisition and distribution of contraceptives were obstacles. CONCLUSION: Lessons learned from the implementation of Z-CAN from the perspective of physicians and other staff can be used to work towards sustainable contraceptive services in Puerto Rico and inform other contraception-access programs' design and implementation strategies.

Engaging Community-Based Organizations to Address Barriers in Public Health Programs: Lessons Learned From COVID-19 Vaccine Acceptance Programs in Diverse Rural Communities.

Factors such as geography, community hesitancy, the political landscape, and legislative efforts to limit public health authority have contributed to a disproportionate number of COVID-19 infections and deaths in US rural communities. Community-based organizations are trusted entities that provide social and educational services in the communities where they live and have proven to be effective public health partners in response to the COVID-19 pandemic. Recognizing the unique challenges faced by rural communities, coupled with higher rates of vaccine hesitancy, the CDC Foundation awarded grants to 21 community-based organizations serving rural communities in 7 Midwest states to support the equitable uptake and distribution of COVID-19 vaccines. In this case study, 2 grantees, the Missouri Center for Public Health Excellence and the Hmong American Center, provide case studies that document their experiences, challenges, and strategies for overcoming barriers during the implementation of COVID-19 vaccine acceptance projects in diverse rural communities. These case studies provide key lessons learned that can be applied to future public health emergency and nonemergency responses to ensure that all members of communities are served well and protected.

Building Global Health Systems Capacity During COVID-19 to Improve Vaccination Access and Reduce Hesitancy: Case Studies in Zambia and Tanzania.

During the COVID-19 pandemic, the CDC Foundation collaborated with implementing partners in Zambia and Tanzania to address challenges related to vaccination access and hesitancy through strategic partnership, technical assistance, and community engagement. These efforts were successful in fostering collaborations among community partners and health authorities and actively engaging the ministries of health. This article describes 2 case studies from Zambia and Tanzania involving different strategies to build health system capacity through projects that improved vaccination access and reduced hesitancy. Such projects illustrate how efforts that strengthen public health and healthcare systems have further positive implications for building localized response systems through context-tailored approaches and building capacity of local healthcare workers. The case studies are examples of public health emergency response projects that successfully increased vaccination access and reduced hesitancy in local communities by rapidly implementing projects to strengthen health system capacity and resilience.

A chemically controlled Cas9 switch enables temporal modulation of diverse effectors.

CRISPR-Cas9 has yielded a plethora of effectors, including targeted transcriptional activators, base editors and prime editors. Current approaches for inducibly modulating Cas9 activity lack temporal precision and require extensive screening and optimization. We describe a versatile, chemically controlled and rapidly activated single-component DNA-binding Cas9 switch, ciCas9, which we use to confer temporal control over seven Cas9 effectors, including two cytidine base editors, two adenine base editors, a dual base editor, a prime editor and a transcriptional activator. Using these temporally controlled effectors, we analyze base editing kinetics, showing that editing occurs within hours and that rapid early editing of nucleotides predicts eventual editing magnitude. We also reveal that editing at preferred nucleotides within target sites increases the frequency of bystander edits. Thus, the ciCas9 switch offers a simple, versatile approach to generating chemically controlled Cas9 effectors, informing future effector engineering and enabling precise temporal effector control for kinetic studies.

ctDNA guided adjuvant chemotherapy versus standard of care adjuvant chemotherapy after curative surgery in patients with high risk stage II or stage III colorectal cancer: a multi-centre, prospective, randomised control trial (TRACC Part C).

BACKGROUND: Circulating tumour DNA (ctDNA) to detect minimal residual disease (MRD) is emerging as a biomarker to predict recurrence in patients with curatively treated early stage colorectal cancer (CRC). ctDNA risk stratifies patients to guide adjuvant treatment decisions. We are conducting the UK's first multi-centre, prospective, randomised study to determine whether a de-escalation strategy using ctDNA to guide adjuvant chemotherapy (ACT) decisions is non-inferior to standard of care (SOC) chemotherapy, as measured by 3-year disease free survival (DFS) in patients with resected CRC with no evidence of MRD (ctDNA negative post-operatively). In doing so we may be able to spare patients unnecessary chemotherapy and associated toxicity and achieve significant cost savings for the National Health Service (NHS). METHODS: We are recruiting patients with fully resected high risk stage II and stage III CRC who are being considered for ACT into the study which uses results from a plasma-only ctDNA assay to guide treatment decisions. Eligible patients are randomised 1:1 to receive ctDNA-guided chemotherapy versus SOC chemotherapy. The primary endpoint is the difference in DFS at 3 years between the trial arms. Secondary endpoints include the proportion of patients in the ctDNA-guided arm who are ctDNA negative post-operatively and receive de-escalated ACT compared to the standard arm, the difference in overall survival (OS), neurotoxicity and quality of life between the arms, and the cost-effectiveness of ctDNA-guided therapy compared to SOC treatment. We hypothesise that using a ctDNA-guided approach to ACT decisions is non-inferior to SOC. Target accrual is 1621 patients over 4 years, which will provide a power of 80% with an alpha of 0.1 to demonstrate non-inferiority with a margin of 1.25 in survival of the ctDNA-guided approach compared to SOC. We anticipate approximately 50 UK centres will participate. The study opened with the Guardant Reveal plasma-only ctDNA assay in August 2022. DISCUSSION: The trial will determine whether ctDNA guided ACT is non-inferior to SOC ACT in patients with fully resected high risk stage II and stage III resected CRC, with the potential to significantly reduce unnecessary ACT and the toxicity associated with it. TRIAL REGISTRATION: NCT04050345.

Impact of a Health Communication Campaign on Uptake of Contraceptive Services during the 2016-2017 Zika Virus Outbreak in Puerto Rico.

The Zika Contraception Access Network (Z-CAN) was established during the 2016-2017 Zika virus outbreak in Puerto Rico as a short-term emergency response program providing client-centered contraceptive counseling and same-day access to the full range of reversible contraceptive methods at no cost to women wishing to delay pregnancy. An evidence-based communication campaign, Ante La Duda, Pregunta (ALDP), was launched to encourage utilization of Z-CAN services. We assessed the effectiveness of campaign tactics in increasing awareness of Z-CAN among women in Puerto Rico. Data on campaign exposure and awareness were obtained through a self-administered online survey approximately two weeks after an initial Z-CAN visit, while the number of searches for participating clinics were obtained from monitoring the campaign website. Findings demonstrated that the most common ways survey respondents learned about Z-CAN were through friends or family (38.3%), social media (23.9%), a clinical encounter (12.7%), and website (11.7%). Nearly two-thirds (61.1%) of respondents had heard of the ALDP campaign. Over the campaign's duration, there were 27,273 searches for Z-CAN clinics. Findings suggest that evidence-based communication campaigns may increase awareness of needed public health services during emergencies. Word of mouth, social media, and digital engagement may be appropriate communication tactics for emergency response mobilization.

A Prospective Cohort Protocol for the Remnant Investigation in Sepsis Study.

BACKGROUND: Sepsis is a common and deadly syndrome, accounting for more than 11 million deaths annually. To mature a deeper understanding of the host and pathogen mechanisms contributing to poor outcomes in sepsis, and thereby possibly inform new therapeutic targets, sophisticated, and expensive biorepositories are typically required. We propose that remnant biospecimens are an alternative for mechanistic sepsis research, although the viability and scientific value of such remnants are unknown. METHODS AND RESULTS: The Remnant Biospecimen Investigation in Sepsis study is a prospective cohort study of 225 adults (age ≥ 18 yr) presenting to the emergency department with community sepsis, defined as sepsis-3 criteria within 6 hours of arrival. The primary objective was to determine the scientific value of a remnant biospecimen repository in sepsis linked to clinical phenotyping in the electronic health record. We will study candidate multiomic readouts of sepsis biology, governed by a conceptual model, and determine the precision, accuracy, integrity, and comparability of proteins, small molecules, lipids, and pathogen sequencing in remnant biospecimens compared with paired biospecimens obtained according to research protocols. Paired biospecimens will include plasma from sodium-heparin, EDTA, sodium fluoride, and citrate tubes. CONCLUSIONS: The study has received approval from the University of Pittsburgh Human Research Protection Office (Study 21120013). Recruitment began on October 25, 2022, with planned release of primary results anticipated in 2024. Results will be made available to the public, the funders, critical care societies, laboratory medicine scientists, and other researchers.

Association Between Preexisting Heart Failure With Reduced Ejection Fraction and Fluid Administration Among Patients With Sepsis.

Importance: Intravenous fluid administration is recommended to improve outcomes for patients with septic shock. However, there are few data on fluid administration for patients with preexisting heart failure with reduced ejection fraction (HFrEF). Objective: To evaluate the association between preexisting HFrEF, guideline-recommended intravenous fluid resuscitation, and mortality among patients with community-acquired sepsis and septic shock. Design, Setting, and Participants: A cohort study was conducted of adult patients hospitalized in an integrated health care system from January 1, 2013, to December 31, 2015, with community-acquired sepsis and preexisting assessment of cardiac function. Follow-up occurred through July 1, 2016. Data analyses were performed from November 1, 2020, to August 8, 2022. Exposures: Preexisting heart failure with reduced ejection fraction (≤40%) measured by transthoracic echocardiogram within 1 year prior to hospitalization for sepsis. Main Outcomes and Measures: Multivariable models were adjusted for patient factors and sepsis severity and clustered at the hospital level to generate adjusted odds ratios (aORs) and 95% CIs. The primary outcome was the administration of 30 mL/kg of intravenous fluid within 6 hours of sepsis onset. Secondary outcomes included in-hospital mortality, intensive care unit admission, rate of invasive mechanical ventilation, and administration of vasoactive medications. Results: Of 5278 patients with sepsis (2673 men [51%]; median age, 70 years [IQR, 60-81 years]; 4349 White patients [82%]; median Sequential Organ Failure Assessment score, 4 [IQR, 3-5]), 884 (17%) had preexisting HFrEF, and 2291 (43%) met criteria for septic shock. Patients with septic shock and HFrEF were less likely to receive guideline-recommended intravenous fluid than those with septic shock without HFrEF (96 of 380 [25%] vs 699 of 1911 [37%]; P < .001), but in-hospital mortality was similar (47 of 380 [12%] vs 244 of 1911 [13%]; P = .83). In multivariable models, HFrEF was associated with a decreased risk-adjusted odds of receiving 30 mL/kg of intravenous fluid within the first 6 hours of sepsis onset (aOR, 0.63; 95% CI, 0.47-0.85; P = .002). The risk-adjusted mortality was not significantly different among patients with HFrEF (aOR, 0.92; 95% CI, 0.69-1.24; P = .59) compared with those without, and there was no interaction with intravenous fluid volume (aOR, 1.00; 95% CI, 0.98-1.03; P = .72). Conclusions and Relevance: The results of this cohort study of patients with community-acquired septic shock suggest that preexisting HFrEF was common and was associated with reduced odds of receiving guideline-recommended intravenous fluids.