bluuebunny/crossref_metadata_2025 huggingface dataset:
500 Quantification of the HIV reservoir in the gut-associated lymphoid tissue
Authors: Cossarini Francesca , D’Souza Darwin , Roudko Vladimir , Aberg Judith , Mehandru Saurabh
Published: April 2024

OBJECTIVES/GOALS: The major obstacle to an effective cure or remission for HIV infection is the integration of HIV into the genome of long-lived resting cells which constitute the so-called viral reservoir. With this study we want to elucidate the changes of the gut-associated HIV reservoir at different stages of viral suppression METHODS/STUDY POPULATION: Recent studies have shown that after long-term (>7 years) clinical suppression of peripheral HIV RNA, the circulating viral reservoir does not seem to decline further and, in fact may expand. The gastrointestinal associated lymphoid tissue (GALT) harbors by far the largest fraction of the latently infected cells, however not much is known about its changes over time.We thus quantified the HIV viral reservoir in the GALT by identifying HIV viral transcripts via 10X single-cell RNA sequencing at two GALT-sites in five PWH and compared the amount of HIV RNA found in the group of PWH with early (< 7years) vs late (> 7years) peripheral virological suppression (plasma HIV RNA <20copies/mL). RESULTS/ANTICIPATED RESULTS: Study participants had been diagnosed with HIV infection for a median (IQR) of 31 (32-34) years and had consistently undetectable peripheral blood HIV RNA for the previous 8 (4-15) years. In PWH with consistent viral suppression < 7yrs, 4 (2-6) HIV transcripts were identified in the ileum and 25 (13 – 38) in the colon. In PWH with consistent viral suppression > 7yrs, 0 (0-4) HIV transcripts were identified in the ileum and 7 (14-11) in the colon. Based on these preliminary results we plan to expand our cohort and confirm these results using Proviral DNA quantification. We anticipate that the viral decay in the GALT will follow a slower dynamic than what has been reported for the peripheral blood achieving a steady state after more than 7 years of peripheral viral suppression. DISCUSSION/SIGNIFICANCE: Despite the remarkable progress the survival and quality of life of PWH, after forty years from its first discovery, HIV infection remains uncurable. Considering its critical role, efforts are needed to better understand the dynamics of the GALT-associated HIV reservoir.
569 Investigating the Role of FOXA2 During the Transition to Neuroendocrine Prostate Cancer
Authors: Garner Richard , Brady Nicholas , Dunmore Kate , Singh Richa , Bagadion Alyssa , Sboner Andrea , Elemento Olivier , Robinson Brian , Beltran Himisha , Rickman David S.
Published: April 2024

OBJECTIVES/GOALS: The goal of this project is to characterize the efficacy of FOXA2 as a potential biomarker for patients with metastatic castrate-resistant prostate adenocarcinoma (CRPC) before transitioning to neuroendocrine prostate cancer (NEPC). NEPC currently has no therapeutic options and poor mechanistic understand of its origins. METHODS/STUDY POPULATION: In our study, we have utilized a multi-omics approach to characterize the potential efficacy of FOXA2 as a prognostic biomarker in numerous patient-derived castrate-resistant prostate cancer (CRPC) models. We have performed ATAC-, ChIP-, RNA-seq and proteomics to fully characterize where FOXA2 is binding genome-wide, how FOXA2 alters chromatin accessibility dynamics, identify regulatory gene targets of FOXA2, and identify FOXA2 protein-protein interactors. We have supported these findings using publicly available data from independent CRPC and NEPC patient cohorts and prostate cancer cell models. RESULTS/ANTICIPATED RESULTS: Our findings show that FOXA2 overexpression suppressed androgen signaling and promoted progression to a NEPC phenotype under short- and long-term androgen deprivation conditions, respectively. Further, FOXA2 redirected the chromatin accessibility landscape to be consistent with an NEPC gene expression program, including increased chromatin accessibility for key NEPC transcription factors. FOXA2 ChIP-seq showed FOXA2 to be bound at known NEPC driver genes and epigenetic modifiers across multiple stages of prostate cancer progression. Lastly, we discovered that FOXA2 physically interacts with key NEPC TFs and epigenetic regulators, suggesting that these FOXA2 physical interactions are required for NEPC progression. DISCUSSION/SIGNIFICANCE: This project will determine the efficacy of FOXA2 as a biomarker in advanced prostatecancer samples, which will translate as a potentially useful tool for clinicians to use for treatment of advanced prostate cancer patients.
504 Dihydroxyacetone, a combustion of electronic cigarettes, promotes cardiac-specific injury through metabolic and mitochondrial imbalances
Authors: Hernandez Arlet , Gwin M , Wiggins LA , Bryant H , Vasilyev M , Dal Zotto VL , Bates ML , Schuler M , Gassman NR
Published: April 2024

OBJECTIVES/GOALS: Electronic cigarettes have become increasingly popular, with various combustion products generated in the process. Dihydroxyacetone (DHA), a carbohydrate made during the heating process. Exposures may reach high micromolar to low millimolar doses of DHA per day and no studies have been done to understand the effects of DHA in the heart. METHODS/STUDY POPULATION: Here, we examine if DHA contributes to these using rat cardiomyocytes, H9c2 cells, and rat cardiac tissues to DHA evaluating metabolic and mitochondrial effects. Using the cells, we will investigate metabolic and mitochondrial pathways using Seahorse, protein expression changes in nutrient sensing pathways, and understand dose-dependent effects of DHA in the heart. Metabolite pools will also be evaluated to understand the changes promoted by DHA. Oxidative stress as previously observed in other cell models will also be measured. Key findings in the cardiac cells will be investigated in the cardiac tissues exposed to DHA. RESULTS/ANTICIPATED RESULTS: We have previously shown DHA induces oxidative stress, metabolic changes, and mitochondrial dysfunction in various cell line models. Interestingly, these effects are highly cell-type dependent. E-cigarettes are known to have toxic cardiac effects, including arterial stiffness, endothelial dysfunction, vascular injury, and oxidative stress. Changes in glycolytic, fatty acid synthesis, and the citric acid cycle enzymes and metabolites were found in the H9c2 cells. We also observed increased mitochondrial ROS and fuel changes due to DHA exposure. In DHA exposed cardiac tissues, we observed oxidative stress and mitochondrial fission and fusion dynamics altered. DISCUSSION/SIGNIFICANCE: These data suggest further study at physiologically relevant doses is warranted to understand how DHA inhaled impacts the long-term health of vapers. As well as the regulation of DHA in e-cigarettes as it has been deemed as safe for topical applications and warned against inhalation.
509 Regulatory Lens of a QA/QC Project Manager
Authors: Bennett Rachel , Caldwell Christine Sego
Published: April 2024

OBJECTIVES/GOALS: The primary purpose of the QA/QC Project Manager (PM), appointed under the NCATS UL1 administrative supplement award, is to facilitate quality and timely NCATS prior approval submissions preventing study start delays. Other goals include supporting these projects’ IRB applications and monitoring to ensure data quality and compliance. METHODS/STUDY POPULATION: At the Indiana CTSI, the QA/QC PM is assigned to the Regulatory Knowledge and Support program (RKS) and functions as a unique regulatory service provider. Through monitoring, auditing, and personalized consultations, the IN CTSI QA/QC PM provides study teams with regulatory, GCP, and other compliant study conduct insights while managing NCATS prior approval and RPPR submission quality and timeliness. In contrast to many CTSAs, this role is uniquely situated within RKS and provides QA/QC support through a regulatory lens. The Indiana CTSI QA/QC PM serves on the CTSA QA/QC Lead Team collaborating with NCATS and other CTSA QA/QC personnel. The Lead Team engages with NCATS to host monthly/quarterly meetings and participate in a discussion forum of NCATS and other CTSA QA/QC personnel. RESULTS/ANTICIPATED RESULTS: Not all CTSAs employ the QA/QC PM as regulatory support and the role and skill sets at each CTSA vary, yet the collaborative nature of these individuals across the CTSAs facilitates sharing of resources and knowledge. While prior approval and RPPR submissions vary widely, the QA/QC PMs can rely on their counterparts for guidance complying with the same regulations and policies within unique research settings and institutional nuances. The IN CTSI QA/QC PM, in collaboration with the QA/QC Lead Team, provided quality assurance revisions to the NCATS prior approval instructions which were adopted and published by NCATS January 2022 for implementation at all CTSAs. Ongoing, quality control efforts are accomplished through education, monitoring, and regulatory consultations. DISCUSSION/SIGNIFICANCE: As the research environment evolves, the QA/QC PM responsibilities shift in response to needs within RKS and NCATS. The versatility of the position enables QA/QC to occur at all stages of a study. QA/QC strategies aim to facilitate communication, quality NCATS prior approval and RPPR submissions, and compliance with proposed study conduct.
523 Measuring the Impact of an Operations Manager on a Study Team
Authors: Martusewicz Zoe J. , Cottler Linda B.
Published: April 2024

OBJECTIVES/GOALS: Having a strong leader in the form of an operations manager is crucial to lead and motivate teams. They're an additional set of problem-solving eyes to identify and address challenges. This poster demonstrates coordination techniques involved to work with teams of researchers, ensuring effective communication, deadlines met, and impact. METHODS/STUDY POPULATION: Stories, course corrections, publications, presentations and other metrics related to various projects and studies in the Lab will show such as the National Drug Early Warning System (NDEWS), Program to Alleviate National Disparities in Ethnic and Minority Immunizations in the Community (PANDEMIC, CDC), Polysubstance Use Study (PSU), and All of Us Consortium of CTSA/PACER Community Network. Additional data will be collected with project managers and students from each respective project to evaluate the effectiveness of an Operations Manager on the team across all studies in Our Lab. RESULTS/ANTICIPATED RESULTS: By sharing these results, showing the benefit to having an Operations Manager and the stories collected and shared for this poster, course corrections will set an example for future operations managers and their teams to continue to optimize efforts. Tis Could be a Special Interest Group (SIG)-related effort that would facilitate success to many labs. DISCUSSION/SIGNIFICANCE: Operations management focuses on effectively managing resources, ensuring that we meet deadlines, reduce delays, and maximize team productivity. By sharing our experiences, this can be a more standard practice among research labs to have smooth operations, and increase overall reputation of our team among those in the field.
524 Navigator: Providing a foundation for cross team collaboration and custom research service through the CTSA Hub
Authors: Swartz Kitt , Honoré Matt , Cook Kimberly , Funderburgh Heidi , Morris Cynthia , Ellison David
Published: April 2024

OBJECTIVES/GOALS: The Oregon Clinical and Translational Research Institute (OCTRI) Clinical Research Navigator program provides a single point of entry for clinical and translational research services, support, advice and guidance. We provide data to illustrate the Navigator model at OHSU and examine continued opportunities to optimize research resources. METHODS/STUDY POPULATION: Requests and activities performed by the OCTRI Navigator program, staffed by 3 FTE (2 Assistant Navigators and 1 Assistant Director) were analyzed. Navigator receives requests through multiple methods: a digital form (REDCap®), email, phone calls. Requests for services and support include focused need for a core or a broad request for multiple services for start-up: informatics, the clinical and translational research center, regulatory knowledge and support, recruitment, qualitative methods, community research, biostatistics or broad consultations. Requests are tracked in SPARCRequest. Navigator also supports wayfinding to institutional resources outside of the CTSA, matchmaking for sponsors seeking investigators, and serves as a connector and facilitator across programs. RESULTS/ANTICIPATED RESULTS: OCTRI Clinical Research Navigator triaged an average of 964 research requests for 613 projects with 388 unique investigators annually between 2018-2022. Navigator also fields more than 80 calls each year that are unrelated to CTSA projects. Project requests are examined to illustrate trends in projects requesting multiple services and display how Navigator simplifies project intake and connects researchers to resources they may have not recognized they needed. Project attributes including funding type and funding status are included in this review. DISCUSSION/SIGNIFICANCE: CTSA resources are essential to the infrastructure available to researchers. While absolute numbers of requests provide little insight into the impact each CTSA hub may have, the timing and clustering trends of projects with multiple program requests shows how a combination of technology and experienced staff can efficiently support researchers.
547 The viscous and fermentability properties of dietary fiber impact on chronic kidney disease-mineral and bone disorder
Authors: Biruete Annabel , Chen Neal X. , Srinivasan Shruthi , O’Neill Kalisha , Siles Samantha , Gallant Kathleen Hill , Moe Sharon M.
Published: April 2024

OBJECTIVES/GOALS: Dietary fiber has been used in other clinical populations to improve mineral disorders, but there is limited data in chronic kidney disease, despite the high prevalence of mineral and bone disorder (known as CKD-MBD). Our objective was to evaluate the effect of dietary fiber based on viscosity and fermentability on CKD-MBD outcomes. METHODS/STUDY POPULATION: 22-week-old male CKD rats (mild-to-moderate CKD) were randomly assigned to receive one of four fiber treatments (10% w/w each) based on fermentability and viscosity: 1) Cellulose (-fermentability, -viscosity), 2) Inulin (+fermentability, -viscosity), 3) Psyllium husk (-fermentability, +viscosity), or 4) Pectin (+ fermentability, +viscosity). Treatments lasted 10 weeks, and rats were euthanized at 32 weeks of age (kidney failure). Rats were placed in metabolic cages for 3 consecutive days the last week before euthanasia for mineral balance. At euthanasia, blood, tibia, heart, and aorta were collected for CKD-MBD assessment. Additional tissues collected included kidneys and all intestinal segments. RESULTS/ANTICIPATED RESULTS: Our preliminary data indicates that weight trajectories and survival were similar between treatment groups. At 33 weeks of age, kidney weight index (an indirect measurement of kidney function as this animal model develops polycystic kidneys) was lower in the psyllium-treated rats compared to all of the other treatments. Plasma phosphorus was lower with Psyllium and Pectin compared to Cellulose-treated rats. Left ventricular mass index was lower in the Inulin, Psyllium, and Pectin-treated rats compared to the Cellulose-treated rats. Ongoing tissue analyses include biochemical markers of mineral and bone metabolism (parathyroid hormone, fibroblast growth factor-23, and phosphorus balance), bone parameters (dynamic histomorphometry and microCT), and cardiovascular calcification. DISCUSSION/SIGNIFICANCE: Our preliminary data indicate that dietary fiber based on fermentability and viscosity impacts CKD-MBD outcomes and may be an innovative, low-cost intervention that can be trialed in people with CKD for the prevention and treatment of CKD-MBD.
30 Characteristics of Infant Emergency Department Utilization
Authors: Howard Mary Beth , Ryan Leticia M. , Badaki-Makun Oluwakemi
Published: April 2024

OBJECTIVES/GOALS: Frequent utilizers of emergency departments (ED) make up a substantial share of overall ED use. Within pediatric emergency departments (PED), infants represent an age group that make up a disproportionate share of PED visits. The objective was to compare patterns of PED use for children less than 1 year of age by visit frequency and resource utilization. METHODS/STUDY POPULATION: Retrospective cohort study of infants less than 1 year presenting to 5 EDs in one health system over a 5-year period, with a 365-day follow-up after each index visit. Patient characteristics (age, sex, race/ethnicity, presence of chronic condition) and visit characteristics (arrival day/time, acuity level, disposition, testing (labs and radiographs, medications) were assessed. The relationship between patient and visit characteristics with utilization and repeat visits was assessed using multivariable regression. RESULTS/ANTICIPATED RESULTS: A total of 20,620 patients with 33,127 ED visits during study timeframe. Thirty three percent (n=6842) had more than one visit in a year; 3964 (19.2%) had two visits, 1542 (7.5%) had three visits, and 1336 (6.5%) had 4 or more visits. Across all visits, over half (52%) were low acuity. The most common diagnoses were respiratory diseases (27%), systemic states (including fever, viral illness, 23%), and gastrointestinal diseases (15%). These diagnoses remained the most common for those with 1, 2, 3, and ≥ 4 ED visits during follow up. As ED visit frequency increased, there was an increase in percentage of children who were older, non-Hispanic Black, and triaged as low acuity. Infants with ≥ 4 ED visits were more likely to be without a chronic condition, have no medications or testing ordered, and be discharged. DISCUSSION/SIGNIFICANCE: There was high ED utilization for those without chronic conditions who were least likely to need medications, testing, and hospital admission. With increasing attention paid to high-utilization in healthcare, it is important to assess why infants use the ED at high rates and develop systems to improve high value care while decreasing resource burden.
564 Platelets and Leukocytes Interact to Modulate Inflammation in Patients with CKD
Authors: Jain Nishank , Kore Rajshekhar , Arthur John M , Ware Jerry , Pathak Rupak
Published: April 2024

OBJECTIVES/GOALS: Platelets interact with leukocytes in the circulation to modulate inflammation in chronic diseases. In previous clinical study, we showed that platelet leukocyte interaction is reduced in the circulation of patients with CKD. Preclinical studies are needed to show whether these findings are a precursor to or a result of CKD. METHODS/STUDY POPULATION: We used mouse models (wild type and platelet-defect) and induced CKD with intraperitoneal cisplatin injections. We measured platelet leukocyte interactions before and after CKD induction in the two models. RESULTS/ANTICIPATED RESULTS: We found platelet-leukocyte interaction to reduce after CKD induction in both wild type and platelet-defect mice. This coincided with a pro-inflammatory state in these mice, as measured by serum TNFalpha levels. Specifically, pro-inflammatory state was exacerbated in CKD of mice with platelet-defects compared to the wild type. DISCUSSION/SIGNIFICANCE: These findings recapitulate translational findings in human CKD samples and confirm that CKD state results in reduced platelet-leukocyte interactions in the circulation, and this change imparts a pro-inflammatory state in the CKD state.
51 School Shootings and Mental Health in the United States
Authors: Rencken Camerin , Ellyson Alice , Rhew Isaac , Davis Carol A. , Rowhani-Rahbar Ali
Published: April 2024

OBJECTIVES/GOALS: It is estimated that 357,000 children have experienced a school shooting since 1999, yet due to limitations in the firearm violence field broadly, the sequalae are not well understood. The objective of this work is to examine the mental health impacts of school shootings, providing insight into the lasting effects of firearm violence on our communities. METHODS/STUDY POPULATION: We will first conduct a quasi-experimental study using controlled interrupted time series with repeated cross-sectional data to assess school shootings’ impact on US mental health. School shooting data is from the K-12 School Shooting Database, and mental health data will be collected via the Behavioral Risk Factor Surveillance System. Second, we will conduct focus groups with community organizations, school administrators, and the public. Interview guides will be developed to explore the mental health impacts of school shootings, to guide the quantitative results interpretation, and assess educational materials’ usefulness. Qualitative analysis will occur in NVivo software with codebook refinement through thematic analysis. Results will be triangulated through convergence coding. RESULTS/ANTICIPATED RESULTS: This research is situated within the context of the pervasive mental health challenges in the US, where mental illness poses significant health, social, and economic burdens. Thus, we anticipate finding an association between school shootings and decreased self-reported mental well-being among US adults. Literature suggests that there may be a stronger association among specific subgroups, such as parents with school-aged children or individuals living in close proximity to such incidents. We expect to find heterogeneity in the effect estimate based on school shooting attributes, such as the number of casualties. Through focus groups, we anticipate furthering our comprehension of the broad-ranging effects of school shootings on less quantifiable outcomes and the unique trajectories of recovery. DISCUSSION/SIGNIFICANCE: This project will contribute needed information on the impact of school shootings and mental health and assist in reducing the frequency and impact of school shootings. Furthermore, we aim to extend our findings beyond the scientific community, translating them into educational resources advocating for policy and public health interventions.
61 A Multifaceted Approach to Improving Fish Farming in Kenya’s Lake Victoria Region
Authors: Fiorella Kathryn , Teplitz Eric , Getchell Rodman , Gonzalez Grace E.
Published: April 2024

OBJECTIVES/GOALS: This project adopts a multifaceted approach to improving aquaculture management practices in Kenya’s Lake Victoria region by identifying fish pathogens, measuring algal toxin levels in commonly consumed fish, surveying fish farming practices, and educating the public. METHODS/STUDY POPULATION: Limited existing data on the state of floating cage culture in Kenya influenced our decision to begin this portion of the project with a brief literature review of potential Nile tilapia pathogens. Databases were screened for mention of disease in either wild or caged Nile tilapia, with emphasis given to those in Lake Victoria. Results were compiled into a spreadsheet and analyzed for frequently occurring pathogens. The next portion involved creating an interview style survey to assess current cage culture management practices in the region. Editing was done to ensure questions remained unbiased, non-leading, culturally sensitive, multilingual and relevant to the situation. Data went through a quality control screening and analysis was conducted through the R programming language. RESULTS/ANTICIPATED RESULTS: Beginning with mortality, of the 93 farms surveyed, data analysis revealed that there is a higher probability that farms will have a mortality of approximately 20%, over the course of a production cycle. For biosecurity and fish health practices, data shows that 97% of farms do not disinfect scooping nets or other fish handling materials when moving from one cage to another. During the 2022-2023 production cycle, 44% of farms experienced fish kills of over 50 fish. 73% of the 93 farms do not contact any organization when a fish kill occurs. In a qualitative answer, it also appears that many farm workers dispose of their dead fish within the lake, feed it to livestock or dogs, or eat it. Algae blooms have been experienced at 80% of the farms surveyed and 43% of farms say they have seen fish gasping at the surface for air. DISCUSSION/SIGNIFICANCE: While farms are implementing good management practices in the areas of cage design, stocking, and feeding practices, there is room for improvement in fish health, biosecurity, and managing algal blooms. The findings provide insight into the areas that should be considered when taking action to improve the welfare of the region.
81 A rapid-cycle application of the Consolidated Framework for Implementation Research allows timely identification of barriers and facilitators to implementing the World Health Organization’s Emergency Care Toolkit in Zambia
Authors: Burkholder Taylor , Dixon Julia , Broccoli Morgan , Chenga Natasha , Chibesakunda Patricia , Kunda Winnie , Mwanza Kephas E , Nonde James , Chiluba Mwiche
Published: April 2024

OBJECTIVES/GOALS: Implementation science evaluations are often too time-intensive to provide actionable feedback during implementation, suggesting the need for more agile methods. We present an evaluation of the World Health Organization’s Emergency Care Toolkit implementation in Zambia using rapid qualitative methods to provide timely feedback. METHODS/STUDY POPULATION: We evaluated the implementation of the Emergency Care Toolkit in eight general and referral hospitals in Zambia in 2023 using a rapid-cycle, qualitative template analysis approach grounded in the Consolidated Framework for Implementation Research (CFIR). We gathered qualitative data from operational field notes, focus groups, and key informant interviews of administrators, clinicians, nurses, and support staff in all eight hospitals in Zambia. We parsimoniously applied CFIR constructs and tool-specific codes, focused on barriers and facilitators, to allow for rapid but comprehensive cross-case analysis. The results were used to generate a matrix of stakeholder-relevant, plain-language barriers and facilitators for each tool. RESULTS/ANTICIPATED RESULTS: We completed eight site visits with focus groups and interviews following initial implementation in September 2023 to gather firsthand knowledge related to implementation of the Toolkit. The CFIR-focused coding accelerated analysis by centering on barriers and facilitators for each tool while maintaining a comprehensive evaluation framework. Summary tables of barriers and facilitators were easily interpreted by lay stakeholders. Visualization in tables allowed for identification of common themes across tools and hospitals, making comprehensive recommendations to the implementation and dissemination process quickly possible. We anticipate the study findings will empower implementing partners to make timely, actionable improvements. DISCUSSION/SIGNIFICANCE: Rapid-cycle qualitative implementation evaluations allow for rigorous yet timely feedback on the implementation process compared to traditional methods. This efficient strategy is particularly important in resource-constrained environments where inefficient implementation wastes limited resources and create delays that cost lives.
85 Investigations of Clinical and Translational Science Roadblocks: a Survey of a Private Medical School and a Large Public University
Authors: Sippel Katherine H. , Kanwal Fasiha , Amos Christopher I. , Liao Gloria , Zhu Dakai , Minard Charles , Neuhauser Claudia , Dickinson Mary , Beech Bettina M.
Published: April 2024

OBJECTIVES/GOALS: Clinical and translational science needs to address roadblocks to translational processes. We conducted a survey at two institutions, a private medical school and a large public university, to understand the frequency and distribution of barriers and roadblocks to research. METHODS/STUDY POPULATION: We reviewed the literature to compile a pool of barriers and roadblocks and convened a panel of relevant stakeholders to develop a 20-item questionnaire. Survey respondents were asked to select and prioritize the five leading clinical and translational roadblocks, provide information regarding their academic degrees and rank/position, complete open-ended items regarding their areas of research, and optionally add additional remarks in a comment box. The survey was disseminated in August 2022 via REDCap to faculty and staff with active research protocols at Baylor College of Medicine and the University of Houston. RESULTS/ANTICIPATED RESULTS: In total, 227 respondents completed the survey. Their disciplines were basic science (29.5%), translational research (52.9%), clinical research (55.5%), community-engaged research (9.7%), and educational research (9.7%). Respondents identified 1) lack of access to trained research coordinators, 2) lack of understanding about different resources that facilitate research, 3) complex regulatory environment and delays, 4) fragmented infrastructure for administrative and fiscal processes, and 5) inadequate funding for pilot projects to foster new research. Other roadblocks included lack of established community stakeholder partnerships, inadequate access to medical record data, and limited biostatistical support. In the comments, several respondents noted that all items included were important. DISCUSSION/SIGNIFICANCE: Research workforce recruitment/training was the highest priority followed by lack of access to information and administrative bottlenecks. We are building an online portal to increase awareness and simplify access to competency-based training and research services. Initiatives are underway to address other roadblocks.
91 Using the Health Stigma Discrimination Framework for Understanding Stigma in the Context of Sexual Assault
Authors: Vernetti Erin , Flannery Marie , LeBlanc Natalie
Published: April 2024

OBJECTIVES/GOALS: This theory analysis aims to evaluate a middle-range framework, the HSDF1, in the context of sexual assault stigma incorporating the myriad levels within within culture and society through which stigma can occur and be reinforced. METHODS/STUDY POPULATION: Databases: PubMed, CINAHL, EMBASE, Google Scholar, Organization websites, Citation searchesn = 32Mark Risjord’s “Middle-Range Theories as Models: New Criterion for Analysis and Evaluation” (2019) RESULTS/ANTICIPATED RESULTS: The innovative approach of the HSDF guides understanding of sexual assault stigma in a holistic way, incorporating individual and institutional stratum of the phenomenon. Understanding through integration of this theoretical framework alongside current knowledge may more succinctly inform trauma-informed care for survivors, policy, and cultural awareness for nurses, healthcare providers, police, social workers, and myriad others with whom survivors interact. DISCUSSION/SIGNIFICANCE: Applying the HSDF framework to sexual assault stigma could help break down barriers and raise survivors out of stigmatization, affecting population health through reduced negative health sequelae experienced by survivors.
98 The Crosstalk between Mitochondrial Dysfunction and Neurodevelopmental Outcomes in Preterm Infants with Pain/Stress in the NICU
Authors: Zhao Tingting , Chang Xiaolin , Biswas Subrata , Balsbaugh Jeremy , Liddle Jennifer , Chen Ming-hui , Matson Adam , Cong Xiaomei
Published: April 2024

OBJECTIVES/GOALS: Early life pain/stress impacts infants’ neurodevelopmental outcomes. Mitochondrial dysfunction may interface between infants’ stress and neurodevelopment. The study aims to investigate the associations between pain/stress, proteins associated with mitochondrial dysfunction, and neurobehavioral responses in preterm infants. METHODS/STUDY POPULATION: A prospective cohort study was conducted with 33 preterm infants enrolled between September 2017 and July 2022 at two affiliated NICUs in Hartford and Farmington, CT. Daily pain/stress experienced during NICU was documented. At 36-38 weeks post-menstrual age (PMA), neurobehavioral outcomes were evaluated using the NICU Network Neurobehavioral Scale (NNNS) and buccal swabs for Mass spectrometry-based proteomics analysis. Lasso statistical methods were conducted to study the association between protein abundance and infants’ NNNS summary scores. Multiple linear regression and Gene Ontology (GO) enrichment analyses were performed to examine how clinical characteristics and neurodevelopmental outcomes may be associated with protein levels and underlying molecular pathways. RESULTS/ANTICIPATED RESULTS: During NICU hospitalization, preterm premature rupture of membrane (PPROM) was negatively associated with neurobehavioral outcomes. The protein functions, including leptin receptor binding activity, glutathione disulfide oxidoreductase activity, and response to oxidative stress, lipid metabolism, phosphate, and proton transmembrane transporter activity, were negatively associated with neurobehavioral outcomes. In contrast, cytoskeletal regulation, epithelial barrier, and protection function were found to be positively associated with neurodevelopmental outcomes. In addition, mitochondrial dysfunction-related proteins (SPRR2A, PAIP1, S100A3, MT-CO2, PiC, GLRX, PHB2, and BNIPL-2, ABLIM1, UNC45A, Keratins, MUC1, and CYB5B) were found to be associated with neurobehavioral outcomes. DISCUSSION/SIGNIFICANCE: Mitochondrial dysfunction-related proteins were observed to be associated with early life pain/stress and neurodevelopmental outcomes in infants. Buccal proteins could be used to predict potential neurobehavioral outcomes. In addition, individualized skin integrity protection should be provided to preterm infants during their NICU stay.
Addressing the Legal Void: Ensuring Justice for Adult Male Rape Victims in Bangladesh
Authors: Alam Md. Tanvir , Abbas Ahmad , Mutmainnah Iin , Tonny Sabrina Afrin
Published: April 2024

The objective of this study is to explore and determine the legal constraints related to adult male rape and the consequences as they exist in the Bangladeshi criminal justice system. Primary and secondary data sources for the study were derived from the narratives of adult male rape survivors, blood relatives of rape survivors, police officers, and legal practitioners. A snowball sampling technique was applied. The study reveals that there is an absence of a police-support response to adult male rape victims, which is attributable to the lack of a gender-neutral definition of rape. This legal definition vacuum diverts complainants to a range of other futile options, such as submitting a general diary and/or adjacent complaints and seeking assistance from other law enforcement agencies. Finally, this study demonstrates that the landscape of the Bangladeshi criminal justice system is likely to remain unchanged until its legal foundation is better developed.
CONSTITUTIONAL VALUES IN THE COMMON LAW OF OBLIGATIONS
Authors: Sales Philip
Published: April 2024

“Constitutional values” is a term which appears to relate to concepts of what is now called public law. By constitutional values, I mean the basic ideas and interests which structure relations between the individual and the state, and the obligations to which they give rise, which underlie the common law and to which it gives recognition in more or less articulated forms. These are ideas and interests such as liberty, private life, freedom of expression and access to justice. Constitutional values and human rights overlap, but they are not necessarily and always the same, either in content or in effect. In exploring this topic I hope to retrieve and bring to the surface an important aspect of the common law in terms of both private law and public law.
HOW DID HOMER'S TROILUS DIE?
Authors: Beck Bill
Published: April 2024

This article examines ancient depictions of the death of Troilus in art and literature and challenges the widespread belief that the Iliad implies an alternative version of the myth in which Troilus dies in battle. In particular, it argues that the death-in-battle interpretation is both insufficiently supported by the internal evidence and incompatible with the external evidence. Given the evident popularity of the story of Achilles’ ambush of Troilus in the Archaic period, it is hard to avoid the conclusion that the poet of the Iliad knew the story of Troilus’ death by ambush. That the poem's only reference to Troilus does not contradict this story, and possibly even alludes to it, should persuade critics of the strong likelihood that the popular story of Troilus’ ambush at the fountain was also the one in the poet's mind.
Carbon Dynamics of a Coastal Wetland Transitioning to Mangrove Forest
Authors: Yannick D. , Oberbauer S. , Staudhammer C. , Cherry J. , Starr G.
Published: April 2024

Coastal wetlands play a vital role in the global carbon cycle and are under pressure from multiple anthropogenic influences. Altered hydrology and land use change increase susceptibility of wetlands to sea‐level rise, saltwater intrusion, tidal flood events, and storm surges. Flooding from perigean spring tides and storm surges rapidly inundates coastal wetlands with saline waters, quickly surpassing vegetation tolerances, leading to shifts in soil microbial respiration, peat collapse, and plant mortality, followed by establishment of salt‐tolerant vegetation. The Southeast Saline Everglades (SESE) is facing many of these pressures, making it a model system to examine the impacts of ecosystem state transitions and their carbon dynamics. Saltwater flooding from Hurricane Irma (2017) initiated a transitional state, where less salt‐tolerant vegetation (e.g., Cladium jamaicense) is declining, allowing halophytic species such as Rhizophora mangle to colonize, altering the ecosystem's biogeochemistry. We utilized eddy covariance techniques in the SESE to measure ecosystem fluxes of CO2 and CH4 in an area that is transitioning to an alternative state. The landward expansion of mangroves is increasing leaf area, leading to greater physiological activity and higher biomass. Our site was presented initially as a small C source (47.0 g C m−2) in 2020, and by 2022 was a sink (−84.24 g C m−2), with annual greenhouse carbon balance ranging from −0.04 to 0.18. Net radiative forcing ranged from 2.04 to 2.27 W m−2 d−1. As the mangrove landward margin expands, this may lead the area to become a greater carbon sink and a potential offset to increasing atmospheric CO2 concentrations.
Progression from ductal carcinoma in situ to invasive breast cancer: molecular features and clinical significance
Authors: Wang Jing , Li Baizhou , Luo Meng , Huang Jia , Zhang Kun , Zheng Shu , Zhang Suzhan , Zhou Jiaojiao
Published: April 2024

Ductal carcinoma in situ (DCIS) represents pre-invasive breast carcinoma. In untreated cases, 25–60% DCIS progress to invasive ductal carcinoma (IDC). The challenge lies in distinguishing between non-progressive and progressive DCIS, often resulting in over- or under-treatment in many cases. With increasing screen-detected DCIS in these years, the nature of DCIS has aroused worldwide attention. A deeper understanding of the biological nature of DCIS and the molecular journey of the DCIS-IDC transition is crucial for more effective clinical management. Here, we reviewed the key signaling pathways in breast cancer that may contribute to DCIS initiation and progression. We also explored the molecular features of DCIS and IDC, shedding light on the progression of DCIS through both inherent changes within tumor cells and alterations in the tumor microenvironment. In addition, valuable research tools utilized in studying DCIS including preclinical models and newer advanced technologies such as single-cell sequencing, spatial transcriptomics and artificial intelligence, have been systematically summarized. Further, we thoroughly discussed the clinical advancements in DCIS and IDC, including prognostic biomarkers and clinical managements, with the aim of facilitating more personalized treatment strategies in the future. Research on DCIS has already yielded significant insights into breast carcinogenesis and will continue to pave the way for practical clinical applications.
Autophagy dysregulation via the USP20-ULK1 axis in the HERC2-related neurodevelopmental disorder
Authors: Sala-Gaston Joan , Pérez-Villegas Eva M. , Armengol José A. , Rawlins Lettie E. , Baple Emma L. , Crosby Andrew H. , Ventura Francesc , Rosa Jose Luis
Published: April 2024

Sequence variants in the HERC2 gene are associated with a significant reduction in HERC2 protein levels and cause a neurodevelopmental disorder known as the HERC2-related disorder, which shares clinical features with Angelman syndrome, including global developmental delay, intellectual disability, autism, and movement disorders. Remarkably, the HERC2 gene is commonly deleted in individuals with Angelman syndrome, suggesting a potential contribution of HERC2 to the pathophysiology of this disease. Given the known critical role of autophagy in brain development and its implication in neurodevelopmental diseases, we undertook different experimental approaches to monitor autophagy in fibroblasts derived from individuals affected by the HERC2-related disorder. Our findings reveal alterations in the levels of the autophagy-related protein LC3. Furthermore, experiments with lysosomal inhibitors provide confirmation of an upregulation of the autophagy pathway in these patient-derived cells. Mechanistically, we corroborate an interaction between HERC2 and the deubiquitylating enzyme USP20; and demonstrate that HERC2 deficiency leads to increased USP20 protein levels. Notably, USP20 upregulation correlates with enhanced stability of the autophagy initiating kinase ULK1, highlighting the role of HERC2 as an autophagy regulator factor through the USP20-ULK1 axis. Moreover, we show that p38 acts as a modulator of this pathway, since p38 activation disrupts HERC2-USP20 interaction, leading to increased USP20 and LC3-II protein levels. Together, these findings uncover a previously unknown role for HERC2 in autophagy regulation and provide insights into the pathomolecular mechanisms underlying the HERC2-related disorder and Angelman syndrome.
RedOx regulation of LRRK2 kinase activity by active site cysteines
Authors: Trilling Chiara R. , Weng Jui-Hung , Sharma Pallavi Kaila , Nolte Viktoria , Wu Jian , Ma Wen , Boassa Daniela , Taylor Susan S. , Herberg Friedrich W.
Published: April 2024

Mutations of the human leucine-rich repeat kinase 2 (LRRK2) have been associated with both, idiopathic and familial Parkinson’s disease (PD). Most of these pathogenic mutations are located in the kinase domain (KD) or GTPase domain of LRRK2. In this study we describe a mechanism in which protein kinase activity can be modulated by reversible oxidation or reduction, involving a unique pair of adjacent cysteines, the “CC” motif. Among all human protein kinases, only LRRK2 contains this “CC” motif (C2024 and C2025) in the Activation Segment (AS) of the kinase domain. In an approach combining site-directed mutagenesis, biochemical analyses, cell-based assays, and Gaussian accelerated Molecular Dynamics (GaMD) simulations we could attribute a role for each of those cysteines. We employed reducing and oxidizing agents with potential clinical relevance to investigate effects on kinase activity and microtubule docking. We find that each cysteine gives a distinct contribution: the first cysteine, C2024, is essential for LRRK2 protein kinase activity, while the adjacent cysteine, C2025, contributes significantly to redox sensitivity. Implementing thiolates (R-S-) in GaMD simulations allowed us to analyse how each of the cysteines in the “CC” motif interacts with its surrounding residues depending on its oxidation state. From our studies we conclude that oxidizing agents can downregulate kinase activity of hyperactive LRRK2 PD mutations and may provide promising tools for therapeutic strategies.
A two-way street – cellular metabolism and myofibroblast contraction
Authors: Noom Anne , Sawitzki Birgit , Knaus Petra , Duda Georg N.
Published: April 2024

Tissue fibrosis is characterised by the high-energy consumption associated with myofibroblast contraction. Although myofibroblast contraction relies on ATP production, the role of cellular metabolism in myofibroblast contraction has not yet been elucidated. Studies have so far only focused on myofibroblast contraction regulators, such as integrin receptors, TGF-β and their shared transcription factor YAP/TAZ, in a fibroblast-myofibroblast transition setting. Additionally, the influence of the regulators on metabolism and vice versa have been described in this context. However, this has so far not yet been connected to myofibroblast contraction. This review focuses on the known and unknown of how cellular metabolism influences the processes leading to myofibroblast contraction and vice versa. We elucidate the signalling cascades responsible for myofibroblast contraction by looking at FMT regulators, mechanical cues, biochemical signalling, ECM properties and how they can influence and be influenced by cellular metabolism. By reviewing the existing knowledge on the link between cellular metabolism and the regulation of myofibroblast contraction, we aim to pinpoint gaps of knowledge and eventually help identify potential research targets to identify strategies that would allow switching tissue fibrosis towards tissue regeneration.
Registered multi-device/staining histology image dataset for domain-agnostic machine learning models
Authors: Ochi Mieko , Komura Daisuke , Onoyama Takumi , Shinbo Koki , Endo Haruya , Odaka Hiroto , Kakiuchi Miwako , Katoh Hiroto , Ushiku Tetsuo , Ishikawa Shumpei
Published: April 2024

Variations in color and texture of histopathology images are caused by differences in staining conditions and imaging devices between hospitals. These biases decrease the robustness of machine learning models exposed to out-of-domain data. To address this issue, we introduce a comprehensive histopathology image dataset named PathoLogy Images of Scanners and Mobile phones (PLISM). The dataset consisted of 46 human tissue types stained using 13 hematoxylin and eosin conditions and captured using 13 imaging devices. Precisely aligned image patches from different domains allowed for an accurate evaluation of color and texture properties in each domain. Variation in PLISM was assessed and found to be significantly diverse across various domains, particularly between whole-slide images and smartphones. Furthermore, we assessed the improvement in domain shift using a convolutional neural network pre-trained on PLISM. PLISM is a valuable resource that facilitates the precise evaluation of domain shifts in digital pathology and makes significant contributions towards the development of robust machine learning models that can effectively address challenges of domain shift in histological image analysis.