bluuebunny/crossref_metadata_2025 huggingface dataset:
431 Evaluating defective Transcription Coupled-Nucleotide Excision Repair as a mechanism for sensorineural hearing loss in a zebrafish model of Cockayne Syndrome
Authors: Hernandez-Herrera Gabriel , Dugdale Joseph , Schimmenti Lisa A. , Clark Karl , Cotter Ryan
Published: April 2023

OBJECTIVES/GOALS: The characterization of the zebrafish as an animal model for Cockayne Syndrome may guide us towards role of Transcription-Coupled Nucleotide Excision Repair (TC-NER) defects in sensorineural hearing loss. METHODS/STUDY POPULATION: To examine our model, we have developed a zebrafish line with a 9+1 base-pair deletion in the ercc6 gene using TALENs. Mutation has since been confirmed by PCR and subsequent restriction digest with StuI. A series of assays evaluating hair cell morphology, structure and function, as well as ribbon synapse structure, will be used to analyze potential differences between the ercc6 mutant zebrafish line a their wild-type siblings. Additionally, electron microscopy will be used to assess differences in hair cell ultrastructure between the ercc6 mutant zebrafish line a their wild-type siblings. Finally, UVC exposure assays will be used to determine the role TC-NER plays in our novel zebrafish model, and evaluate its potential implications in sensorineural hearing loss. RESULTS/ANTICIPATED RESULTS: We anticipate that biallelic loss of function mutations in the zebrafish ercc6 gene will result in abnormalities in hair cell structure, mechanotransduction, or cell number. Additionally, we anticipate that hair cell ultrastructure and ribbon synapse structure will be impacted by loss of ercc6 expression. DISCUSSION/SIGNIFICANCE: Hearing loss mechanisms associated with defects in TC-NER are yet to be described. We believe our model will provide the tools for a faster and efficient way to carry out Cockayne Syndrome studies while laying the groundwork for the association between TC-NER and hearing loss.
434 Ionizing radiation acoustic imaging (iRAI) for volumetric mapping the dose deep in the liver during radiation therapy
Authors: Zhang Wei , Oraiqat Ibrahim , Litzenberg Dale , Chang Kai-Wei , Hadley Scott , Ba Sunbul Noora , Matuszak Martha M. , Tichacek Christopher , Moros Eduardo G. , Carson Paul L. , Cuneo Kyle C. , Wang Xueding , El Naqa Issam
Published: April 2023

OBJECTIVES/GOALS: The goal of this study was to develop a clinically applicable technique to increase the precision of in vivo dose monitoring during radiation therapy by mapping the dose deposition and resolving the temporal dose accumulation while the treatment is being delivered in real time. METHODS/STUDY POPULATION: Ironizing radiation acoustic imaging (iRAI) is a novel imaging concept with the potential to map the delivered radiation dose on anatomic structure in real time during external beam radiation therapy without interrupting the clinical workflow. The iRAI system consisted of a custom-designed two-dimensional (2D) matrix transducer array with integrated preamplifier array, driven by a clinic-ready ultrasound imaging platform. The feasibility of iRAI volumetric imaging in mapping dose delivery and real-time monitoring of temporal dose accumulation in a clinical treatment plan were investigated with a phantom, a rabbit model, and a cancer patient. RESULTS/ANTICIPATED RESULTS: The total dose deposition and temporal dose accumulation in 3D space of a clinical C-shape treatment plan in a targeted region were first imaged and optimized in a phantom. Then, semi-quantitative iRAI measurements were achieved in an in vivo rabbit model. Finally, for the first time, real-time visualization of radiation dose delivered deep in a patient with liver metastases was performed with a clinical linear accelerator. These studies demonstrate the potential of iRAI to monitor and quantify the radiation dose deposition during treatment. DISCUSSION/SIGNIFICANCE: Described here is the pioneering role of an iRAI system in mapping the 3D radiation dose deposition of a complex clinical radiotherapy treatment plan. iRAI offers a cost-effective and practical solution for real-time visualization of 3D radiation dose delivery, potentially leading to personalized radiotherapy with optimal efficacy and safety.
437 Novel Targeting of Semaphorin 7a-Mediated Survival Signaling in ER+ Breast Cancer to Mitigate Therapy Resistance
Authors: Steinmetz Rachel , Lyons Traci
Published: April 2023

OBJECTIVES/GOALS: Estrogen receptor-positive (ER+) breast cancers (BC) comprise >70% of all BC and cause the most BC-related deaths in women worldwide. Despite available therapies against ER+ BC, recurrence often arises due to development of endocrine therapy resistance. Thus, our goal is to identify novel biomarkers that could serve as therapeutic targets for ER+ BC. METHODS/STUDY POPULATION: We have identified a neuroimmune molecule, Semaphorin 7a (SEMA7A), as a potential biomarker for endocrine therapy resistance, relapse, and poor survival in ER+ BC patients. SEMA7A promotes tumor growth, angiogenesis, epithelial-to-mesenchymal transition, metastasis, and endocrine therapy resistance in our pre-clinical models. Specifically, using in vivo models we have shown that SEMA7A+ MCF7 tumors result in lung metastases that do not respond to fulvestrant, in part via downregulation of ER, posing the need to identify novel, druggable targets for SEMA7A+ ER+ BC patients. SEMA7A is a membrane-bound protein that can inhibit tumor cell death via integrin-mediated PI3K/Akt pro-survival signaling. Thus, we hypothesized that SEMA7A+ ER+ BC may be sensitive to PI3K inhibition in combination with fulvestrant. RESULTS/ANTICIPATED RESULTS: Our preliminary studies confirmed that high SEMA7A expression associates with increased phospho-Akt levels and decreased apoptosis of tumor cells in forced suspension conditions. We also observed that human MCF7 ER+ SEMA7A overexpressing (OE) cells are sensitive to the PI3K (P110ï ¡ï€©ï€ inhibitor, Alpelisib. Also, the combination of Alpelisib with fulvestrant inhibited tumor cell viability in MCF7 cells, which was further enhanced in the SEMA7A OE counterparts. The combination also decreased proliferation and tumor sphere formation. There are currently no therapies that directly target SEMA7A, and here I propose an innovative hypothesis that PI3K inhibition will block SEMA7A signaling. DISCUSSION/SIGNIFICANCE: The role of SEMA7A has been studied in several cancer types, but its function in ER+ BC remains less well understood. In future studies, we will explore the mechanisms by which SEMA7A signals in the cell and to promote tumor cell survival. Delineating these mechanisms will help optimize treatment combinations to improve BC patient survival.
440 Role of angiotensin II in SARS-CoV-2 pathophysiology in a hamster model of COVID-19
Authors: Stanic Branka Milicic , Dugal Nisha , Shults Nataliia , Ji Hong , de Souza Aline M.A. , Hawkins Seth , Wu Xie , Saavedra Juan M. , Sandberg Kathryn
Published: April 2023

OBJECTIVES/GOALS: Hamsters develop COVID-19 similarly to people because the SARS-CoV-2 spike protein binds with high affinity to hamster ACE2 resulting in host cell entry and replication. Our goal was to establish a hamster model that mirrors the lung and brain pathophysiology observed in COVID-19. METHODS/STUDY POPULATION: Hamsters infected with SARS CoV-2 are sacrificed on day 1 and day 6 postinfection. Lung histopathology scoring model was implemented for assessment all pathological relevant changes in the lungs of infected animals on tissue sections stained with hematoxylin and eosin. To quantify the extent and severity of lung pathology, two scoring systems were used: the first evaluated all relevant changes in the lungs of the infected animals and the second evaluated only the pathology associated with the pulmonary vasculature. Percentage of airway affected, airway severity, bronchiolar epithelial hyperplasia, alveoli affected, alveolar severity, type II pneumocyte hyperplasia and vessels affected were analyzed. Total airway score plus total lung alveolar score give lung histopathology score. RESULTS/ANTICIPATED RESULTS: Compared to the control hamster, the hamsters day 1 postinfection, exhibited a higher total airway score [9.00 ± 1.35 vs. 0.25 ± 0.1; p DISCUSSION/SIGNIFICANCE: Establishing this outstanding small animal model of COVID-19 will facilitate studies investigating diagnostics, prognosis and response to treatment in COVID-19 disease. These studies will provide insights that will complement on-going clinical trials on angiotensin type 1 receptor (AT1R) blockers (ARBs) in COVID-19.
441 Southeastern Wisconsin Community-Based Participatory Research using the All of Us Researcher Workbench
Authors: Thareja Suma , Yang Xin , Mia Md Raihan , Upama Paramita Basak , Torres Shary Pérez , Coroft Linda Jackson , McGaughey Kari , Franco Zeno , Whittle Jeff
Published: April 2023

OBJECTIVES/GOALS: Using a community-based participatory research (CBPR) approach, the All of Us (AoU) Wisconsin Froedtert & the Medical College of Wisconsin (MCW) site aimed to improve public understanding of science by creating a Special Interest Group (SIG) of community members and scientists and driving healthcare research relevant to Southeastern Wisconsin. METHODS/STUDY POPULATION: We recruited community members from the AoU advisory boards and scientists from Froedtert & MCW. These SIG participants will converge on a hypothesis that aligns with community priorities and conduct research using the national AoU database, which combines EHR, survey, and whole genome sequencing data. A year of monthly SIG meetings from 2022-2023 will serve as focus group sessions that will be recorded and transcribed. In addition to descriptive statistics on participant demographics and affiliations, we plan to qualitatively characterize transcripts, field notes, and feedback, such as participation satisfaction or reflection on self- and group-driven impact. We will publish alongside all SIG members on both the research outcomes and the SIG-driven process to inform future similar endeavors. RESULTS/ANTICIPATED RESULTS: To date, we held three SIG meetings, which together formed the discover and define stages in the human-centered design framework. We narrowed down SIG membership to 15 consistent participants. Our coalition building conversations emphasized group priorities and future payoffs for independent research and community partnerships. Realizing a crucial need for bioinformaticians to prosecute SIG-driven research questions, we created a data subgroup of trainees and scientists to familiarize themselves with the AoU Researcher Workbench. Through shared decision making during this community engaged process, we are converging on a research direction centering on specific diseases or adverse outcomes with relevance to Southeastern Wisconsin communities and with implications for personalized medicine. DISCUSSION/SIGNIFICANCE: SIG scientists will learn about CBPR and access the AoU database. SIG community members will better understand the research process and share their lessons with local communities. The AoU Wisconsin research team will disseminate findings on the collective research process and empower scientist use of this nationally curated AoU database.
446 To Stay or Not to Stay: Multidisciplinary Collaboration After NSF Funding
Authors: Mohammed S. , Tirrell B. , Davis C. , Zhang T. , Basore C. , Liao X. , Miller G.
Published: April 2023

OBJECTIVES/GOALS: NSF often requires cross-disciplinary team composition to be competitive for funding. To what extent do research teams have multidisciplinary authorships after they win an award, given that awards are not contracts? We examined the quantity and quality of multidisciplinary collaboration of NSF-funded teams before and after receiving their award. METHODS/STUDY POPULATION: Our sample was 150 PIs and Co-PIs (67% male) from 58 NSF-funded EAGER (EArly-concept Grants for Exploratory Research) grants between 2013 and 2019. Using publicly available information, we collected the number of conference papers, publications, and grants PIs/co-PIs produced with each other (all PIs and co-PIs in a team or a partial subset). Based on Ph.D. fields, we also cataloged whether the combination of PIs/co-PI authors on outputs represented unidisciplinary or multidisciplinary collaboration after their NSF award. Multidisciplinary collaboration consisted of multidivisional (Ph.D. disciplines across NSF divisions, e.g., political science and cognitive psychology) or multidirectorate (Ph.D. disciplines across NSF directorates, e.g., psychology and engineering) authorship. RESULTS/ANTICIPATED RESULTS: Of the 74% of PI and co-PI teams who collaborated after their EAGER award, almost 9 out of 10 chose to work with a collaborator from a different discipline, and almost 8 out of 10 chose to work with a researcher from an extremely diverse discipline from their own (e.g., computer science and psychology). Research on interdisciplinary teams largely emphasizes the challenges and problems they face but the current research demonstrated that 90% of the sample chose to continue working together across disciplines after EAGER awards. Therefore, future research should dedicate more attention to the nontangible benefits members receive in interdisciplinary teams. Moreover, quality measures revealed higher H-indices for multidisciplinary than unidisciplinary journals and conferences. DISCUSSION/SIGNIFICANCE: Our archival results revealed that NSF EAGER grants are having their intended effect of being a catalyst for 1) continued multidisciplinary (and especially multidirectorate) collaboration) and 2) high-quality multidisciplinary publication and conference output. These results have contributed to NSF policy changes to reinstate the EAGER grant.
453 Rapid SARS-CoV-2 testing with duplexed recombinase polymerase amplification and a bacteriophage internal control
Authors: Martin Coleman , Bender Andrew T. , Sullivan Benjamin P. , Lillis Lorraine , Boyle David S. , Posner Jonathan
Published: April 2023

OBJECTIVES/GOALS: Current COVID-19 rapid molecular tests require cartridge-reader detection, expensive circuitry, and complex microfluidics making the most accurate tests unavailable to the masses. Here we present a rapid molecular diagnostic leveraging isothermal amplification and paper-based microfluidics for a low-cost ultra-sensitive COVID-19 assay. METHODS/STUDY POPULATION: We designed a reverse transcription recombinase polymerase amplification (RT-RPA) assay for the detection of SARS-CoV-2 and bacteriophage MS2 RNA. RT-RPA is a sequence specific, ultrasensitive, rapid isothermal DNA amplification technique that is well suited to home based testing due to its rapid assay time, robustness, ease of use, and readout options. RT-RPA reagents are added to a tube and incubated at 39Â°C in a fluorometer. Realtime fluorometer data gives results in under 15 minutes. This assay also provides visual detection via lateral flow readout with results in 23 minutes. RESULTS/ANTICIPATED RESULTS: We have developed a rapid multiplexed nucleic acid amplification assay with an internal process control for SARS-CoV-2 using single-pot RT-RPA. We screened 21 primer combinations to select primers that demonstrated excellent performance and target specificity against common respiratory viruses. We demonstrate the ability to multiplex SARS-CoV-2 and MS2 detection, utilizing MS2 as an internal process control for lysis, reverse transcription, amplification, and readout. We show duplexed detection using both fluorescence readout and visual readout using lateral flow strips. Duplexed fluorescence detection shows a limit of detection of 25 copies per reaction. Duplexed lateral flow readout shows a limit of detection of 50 copies per reaction DISCUSSION/SIGNIFICANCE: We developed a duplexed RT-RPA assay for SARS-CoV-2 with fluorescence or lateral flow readout. Our assay does not re-quire expensive reader, circuity, or fluid handling. The low material cost, temperature, and robustness make it ideal for a more accurate home-based COVID-19 diagnostic.
459 Caspase-1 mediated inflammatory response - a critical player in concussive mild traumatic brain injury (mTBI) associated long term pain
Authors: Nguyen Tyler , Talley Sarah , Nguyen Natalie , Cochran Ashlyn G. , Al-Juboori Mohammed , Smith Jared A. , Saxena Saahil , Campbell Edward M. , Obukhov Alexander G. , White Fletcher A.
Published: April 2023

OBJECTIVES/GOALS: Patients who have experienced conjunctive mild traumatic brain injuries (mTBIs) suffer from a number of comorbidities, including chronic pain. Despite extensive studies investigating the underlying mechanisms of mTBI-associated chronic pain, the role of inflammation after mTBI and its contribution to long-term pain are still poorly understood. METHODS/STUDY POPULATION: Given the shifting dynamics of inflammation, it is important to understand the spatial-longitudinal changes and their effects on TBI-related pain. Utilizing a recently developed transgenic caspase-1 luciferase reporter mouse, we characterized the bioluminescence signal evident in both in vivo and ex vivo tissues following repetitive closed head mTBIs. This allowed us to reveal the spatiotemporal dynamics of caspase-1 activation in individual animals over time. Furthermore, we utilize various proteomic and behavioral assays to evaluate the role of caspase-1 mediated inflammation in the development and progression of injury-associated chronic pain. Lastly, by blocking inflammasome caspase-1 activation with a specific inhibitor, we assess its clinical potential as the next therapeutic approach to pain. RESULTS/ANTICIPATED RESULTS: We established that there were significant increases in bioluminescent signals upon protease cleavage in the brain, thorax, abdomen, and paws in vivo, which lasted for at least one week after each injury. Enhanced inflammation was also observed in ex vivo brain slice preparations following injury events that lasted for at least 3 days. Concurrent with the in vivo detection of the bioluminescent signal were persistent decreases in mouse hind paw withdrawal thresholds that lasted for more than two months postinjury. Using MCC950, a potent small molecule inhibitor of NLRP3 inflammasome-caspase 1 activity, we observed reductions in both caspase-1 bioluminescent signals in vivo and caspase-1 p45 expression by immunoblotting and an increase in hind paw withdrawal thresholds. DISCUSSION/SIGNIFICANCE: Overall, these findings suggest that neuroinflammation in the brain following repeated mTBIs is coincidental with a chronic nociplastic pain state, and repeated mTBI-associated events can be ameliorated by a highly specific small molecule inhibitor of NLRP3 inflammasome activation.
469 Electroencephalographic Correlate of Sensory Over-Responsivity in Adults with Chronic Tic Disorders
Authors: Isaacs David A. , Conley Alexander C. , Key Alexandra P. , Cascio Carissa J. , Walker Harrison C. , Wallace Mark T. , Claassen Daniel O.
Published: April 2023

OBJECTIVES/GOALS: To identify an electroencephalographic (EEG) signature of SOR in adults with TS METHODS/STUDY POPULATION: We will recruit 60 adults with CTD and 60 sex- and age-matched healthy controls to complete scales assessing severity of SOR (Sensory Gating Inventory, SGI), tics, and psychiatric symptoms. Subjects will then be monitored on dense-array scalp EEG during sequential auditory and tactile sensory gating paradigms, as such paradigms have been shown to correlate with self-report measures of SOR in other populations. Single-trial EEG data will be segmented into 100-ms epochs and spectrally deconvoluted into standard frequency bands (delta, theta, alpha, beta, gamma) for pre-defined regions of interest. We will conduct between-group contrasts (Wilcoxon rank-sum) of band-specific sensory gating indices and within-group correlations (Spearman rank correlations) between sensory gating indices and SGI scores. RESULTS/ANTICIPATED RESULTS: We hypothesize that, relative to controls, adults with CTD exhibit impaired sensory gating and that extent of impairment correlates with severity of SOR. 14 adults with CTD (9 men, 5 women) and 16 controls (10 men, 6 women) have completed the protocol to date. Within this sample, adults with CTD showed significantly reduced sensory gating compared to controls in frontal (CTD median 0.12 dB (interquartile range -0.15–0.70 dB); control -0.37 dB (-0.80–-0.13 dB); p = 0.01) and parietal (CTD 0.17 dB (-0.08–0.50 dB); control -0.20 dB (-0.43–0.10 dB); p = 0.01) gamma band during the 100-200 ms epoch in the tactile paradigm. No significant between-group differences were evident for the auditory paradigm. Among adults with CTD, multiple sensory gating indices significantly correlated with SGI scores. Enrollment continues. DISCUSSION/SIGNIFICANCE: Results aim to clarify the extent of sensory gating impairment in TS and identify a clinical correlate of neurophysiologic dysfunction in the disorder. Such knowledge has direct implications for identification of candidate neurophysiologic biomarkers, an express goal of the National Institutes of Health.
474 Exploring the Genetic Contribution to Oxidative Stress in Myalgic Encephalomyelitis/Chronic Fatigue Syndrome
Authors: Hampilos Nicholas Henry , Germain Arnaud , Mao Xiangling , Hanson Maureen R. , Shungu Dikoma C.
Published: April 2023

OBJECTIVES/GOALS: Strong evidence has implicated oxidative stress (OS) as a disease mechanism in myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS). The study aim was to assess whether a C>T single nucleotide polymorphism (SNP) (rs1800668), which reduces the activity of glutathione peroxidase 1 (GPX1), is associated with brain OS in patients with ME/CFS. METHODS/STUDY POPULATION: Study population: The study enrolled 20 patients with ME/CFS diagnosed according to Canadian Consensus Criteria, and 11 healthy control (HC) subjects. Genotyping: DNA was extracted from whole blood samples, amplified by PCR, and purified. Sanger sequencing was used for genotyping. 1H MRS: Proton magnetic resonance spectroscopy (1H MRS) was used to measure levels of glutathione (GSH)â€” a primary tissue antioxidant and OS markerâ€” in a 3x3x2 cm3 occipital cortex (OCC) voxel. GSH spectra were recorded in 15 minutes with the standard J-editing technique. The resulting GSH peak area was normalized to tissue water level in the voxel. Statistical Analysis: T-tests were used to compare OCC GSH levels between ME/CFS and HC groups, and between the study’s genotype groups (group 1: CC, group 2: combined TC and TT). RESULTS/ANTICIPATED RESULTS: Clinical characteristics: ME/CFS and HC groups were comparable on age and BMI but not on sex (p = 0.038). Genotype frequencies: Genotype frequencies in the ME/CFS group were 0.55 (CC), 0.25 (TC) and 0.2 (TT); and 0.636 (CC), 0.364 (TC), and 0 (TT) in the HC group. GSH levels: There was a trend-level lower mean OCC GSH in ME/CFS than in HC (0.0015 vs 0.0017; p = 0.076). GSH levels by genotype group interaction: Within the ME/CFS group but not in the combined ME/CFS and HC group or HC group alone, GSH levels were lower in the TC and TT genotypes than in CC genotypes (0.00143 vs 0.00164; p = 0.018). DISCUSSION/SIGNIFICANCE: This study found that the presence of a C>T SNP in GPX1 is associated with lower mean GSH levels and, hence, brain oxidative stress, in ME/CFS patients. If validated in a larger cohort, this finding may support targeted antioxidant therapy based on their genotype as a potentially effective treatment for patients with ME/CFS.
477 Impact of COVID-19 Pandemic on Oral Cleft Services in Puerto Rico
Authors: Victoria Yinayra , Buxo Carmen , Elias Augusto , Debs Natalio
Published: April 2023

OBJECTIVES/GOALS: Evaluate the impact of COVID-19 on oral clefts services including surgical and dental treatments in Puerto Rico. METHODS/STUDY POPULATION: This Observational retrospective cohort study will consider patients 0-21 y/o with CL/P that visited the UPR school of Dental Medicine, Pediatric University Hospital Dr. Antonio Ortiz and ongoing case-control research project Face-Genes. Records to be used are classified as follow: Pandemic (March 15, 2020 to March 15 2022) Pre-pandemic (March 15, 2015 to March 15, 2017) Power analysis (power=0.80 alpha=0.05) will be calculated. Unavailable and incomplete medical records and those that did not attended study clinic during study period will be excluded. Data extraction instrument will be based on previous published study. Descriptive statistics, Chi-square, Odds Ratios at 95% confidence intervals and multiple logistic regression will be estimated. RESULTS/ANTICIPATED RESULTS: We hypothesize that surgical and dental services in Puerto Rico will be adversely impacted because of COVID-19 pandemic. DISCUSSION/SIGNIFICANCE: CL/P are common congenital diseases that require early interdisciplinary attention. Lack of timely care as well as surgery and treatment delays, could be associated with poorer prognosis, increased morbidity and mortality. If there is high risk of dh services during emergency situations, our findings will help to allocate the available resources
478 Incidence and Risk Factors for Comorbidities Following COVID-19 Disease in People Living with HIV
Authors: Castellano Cecilia , Moran Caitlin , Nguyen Minh , Lahiri Cecile ,   Emory University
Published: April 2023

OBJECTIVES/GOALS: COVID-19 disproportionately affects patients with prior health conditions and those living at a lower socioeconomic status. Persons living with HIV (PLWH) are infected with SARS-CoV-2 at a higher rate than seronegative patients. Risk factors and incidence of post-COVID-19 comorbidities in PLWH, specifically, are still unknown METHODS/STUDY POPULATION: We will study PLWH enrolled in the Emory Centers for AIDS Research (CFAR) Registry who receive care at the Grady Ponce de Leon Center in Atlanta, Georgia to 1) investigate the incidence of, and 2) identify risk factors that predispose PLWH to post-COVID-19 comorbidities. All PLWH with documented COVID-19 (by positive SARS-CoV-2 PCR or antigen test) between March 1, 2020, and September 30, 2021, with a clinic visit within 12 months will be included. We will identify comorbidities using problem list diagnoses and ICD9/10 codes. With a predicted sample size of 395, we will use a Cox proportional hazards model for time-to-detection of comorbidity, and bivariate and multivariate logistic regression models to identify predictors of incident comorbidity within 12 months of COVID-19. RESULTS/ANTICIPATED RESULTS: o Previous work demonstrated that in PLWH, age and non-AIDS comorbidities, but not HIV-related factors, were associated with hospitalization for COVID-19 in a dose dependent fashion.18 We anticipate that rate of incident comorbidities will be significantly higher in PLWH after COVID-19 compared to PLWH without a history of COVID-19. We also expect that pre-existing comorbidities including obesity and cardiovascular disease, male sex, Black race, and older age are associated with higher incidence of post-COVID-19 comorbidities in PLWH. When stratifying by organ system, we also anticipate that prior comorbidities of an organ system will predispose patients to later complications of that same system. DISCUSSION/SIGNIFICANCE: By understanding the incidence and risk factors associated with developing post-COVID-19 comorbidities
we can improve guidelines for treatment of groups experiencing the disproportionate impact of co-infection with HIV and SARS-CoV-2.
489 Nasal-derived Extracellular Vesicles (EVs) carry a cargo of antiviral and immunomodulatory molecules
Authors: Corsello Tiziana , Ivanciuc Teodora , Qu Yue , Casola Antonella , Garofalo Roberto P
Published: April 2023

OBJECTIVES/GOALS: The goals of this project are to: i) investigate the cargo such as immune mediators (cytokines) and small non coding RNAs (sncRNAs) of EVs derived from nasopharyngeal secretions (NPS) of children with episodes of viral infections and exposed to SHTS; ii) test the biological activity of EVs released from upper airway mucosa on target/recipient lung cells METHODS/STUDY POPULATION: EVs were isolated from ten NPS samples of children with episodes of acute respiratory infections. EVs were characterized by particle sizing (size and concentration), EV markers, and protein arrays for interferons, cytokines, and other immune mediators content. RNA was extracted from ten samples of NPS-derived EVs by column for next generation high throughput sequencing (NGS) to identify sncRNAs in EVs. In studies currently in progress, EVs will be isolated from RSV-infected human nose organoids (HNOs) cells in air liquid interface (ALI) culture, with or without pre-exposure to tobacco smoke. EVs will be then tested for antiviral activity on recipient RSV-infected lower airway cells. Viral titers will be measured in recipient infected lung cells. RESULTS/ANTICIPATED RESULTS: We isolated EVs from NPS samples and confirmed by immunoblot EV markers CD63 and Alix. The average size of NPS-derived EVs of virus positive and negative patients was 170 nm and 145 nm, respectively. We determined the particles number of EVs, concentrations of IFN-βand IFN-Î»in NPS and NPS-derived EVs of these children. While IFN-βlevels were below the limit of detection in both NPS and NPS-derived EVs of all children, IFN-Î»was detected in NPS and NPS-derived EVs from infected patients, except for the two patients with no viral infections. We extracted RNA from control-, virus infected- and/or SHTS- EVs and found that piR-36511, piR-40926, piR-49645, piR-32679 and piR- 53263 were all significantly enriched in EVs derived from NPS of children exposed to TS compared to those not exposed. DISCUSSION/SIGNIFICANCE: RSV leads to approximately 22,000 hospitalizations of children due to second-hand smoke. A vaccine is not currently available for RSV infection. EVs represent a novel translational approach to target undruggable. Airway mucosa EVs in viral respiratory infection function as antiviral messengers and tobacco smoke impairs the EV antiviral activity.
490 Nursing Professionals’Experiences during the COVID-19 Pandemic in Puerto Rico: A Phenomenological Study
Authors: Irene Lourdes , Díaz Andrea Rodríguez
Published: April 2023

OBJECTIVES/GOALS: The COVID-19 pandemic has impacted nursing frontline professionals. The aims of this study were to explore experiences of nursing professionals in Puerto Rico during the pandemic, examine the impact on their health and provide research development opportunities enhance research capacity. METHODS/STUDY POPULATION: This interpretative phenomenological study recruited graduate nurses who participated in one in-depth semi-structured virtual interviews. Interviews were audio recorded and transcribed. The data analysis process was guided using the following steps: 1. Reading and re-reading, 2. Initial noting, 3. Developing emergent themes, 4. Searching for connections across emergent themes, 5. Moving to the next case, 6. Looking for patterns across cases, and 7. Writing up. In addition, Van Manen's thematic structure of four foundations was used as a complement to guide reflection and interpretation. Faculty and students participated throughout the process. RESULTS/ANTICIPATED RESULTS: Seven nursing professionals’lived experiences caring for Covid-19 patients were gathered. Their ages ranged from 31 to 45 and had worked between 2 and 14 years providing direct care. Themes that emerged from narrations include compassion fatigue, teamwork, working beyond clinical role, and gratification. Nurses expressed dealing with a very difficult situation, fear of being infected , or infecting my family , and working together to get through it and better help patients . Nurses also expressed feelings of anxiety and lack of institutional support. Additionally, the impact of working with patients, feeling good for being there, good or bad and support from families. DISCUSSION/SIGNIFICANCE: Nurses’narrations point to the complexities of their experiences working during the pandemic. They had to transcend usual demands even though they often lacked needed support. We must recognize the value of nursing and reflect upon changes in healthcare that are essential to move nursing forward.
492 Phagocyte heterogeneity and T cell dependence of cellular host defense mechanisms in tuberculosis
Authors: Mathes Tailor , Ronayne Christine , Boyd Tyler
Published: April 2023

OBJECTIVES/GOALS: Phagocytes, diverse cells that ingest material, are the primary cell type infected by Mycobacterium tuberculosis (Mtb) and the executors of protective mechanisms. T cells play a critical role by helping phagocytes control the infection. Understanding the precise T cell-dependent mechanisms by which phagocytic cell types contain Mtb is critical. METHODS/STUDY POPULATION: To determine the impact T cells have on different phagocyte cell populations’ host defense mechanisms, groups of wild–type and T cell deficient TCRa-/- mice were infected with an Mtb strain expressing fluorescent mScarlet protein. At four weeks post-infection, a time when T cell help contributes to control of Mtb, lungs were homogenized and cells sorted based on detection of mScarlet, indicating Mtb-infected cells. Cell suspensions from each mouse background were underwent single-cell RNA sequencing analysis to reveal the heterogenous cellular transcriptional response of different phagocyte populations. RESULTS/ANTICIPATED RESULTS: We found that Mtb-infected phagocytes from wild-type and TCRa-/- mouse lungs contain the same dominant cell phenotypic clusters, but these have different patterns of gene expression. Without T cells, phagocytes are prone to a more inflammatory phenotype. DISCUSSION/SIGNIFICANCE: This will translate fundamental biological data to test the hypothesis that Mtb encounters different environmental stresses exerted by different phagocytic cell types. This work could reveal host intracellular niches that enable bacterial persistence and elucidate new pathways that could be targeted for traditional antibiotic therapies for TB.
508 A Study of Cortical Thickness in Bilingual Children with Reading Disability (Dyslexia)
Authors: Schug Alison , Eden Guinevere F.
Published: April 2023

OBJECTIVES/GOALS: Dyslexia is a common Reading Disability (RD) affecting 7-12% of the population and is associated with less cortical thickness (CT) in bilateral brain regions. However, the interaction between RD and a bilingual experience on CT is unknown, even though bilingualism is also associated with altered CT. METHODS/STUDY POPULATION: We studied 48 Bilinguals assigned to the Typical Reader group based on Oral Reading Recognition Test (ORRT) scores above 90 (avg=107 ± 14), 47 Bilinguals assigned to the RD group based on ORRT scores below 85 (avg=77 ± 5), 45 English Monolingual Typical Readers with ORRT scores above 90 (avg=102 ± 13) and 47 Monolinguals with RD based on ORRT scores below 85 (avg=78 ± 5). Participants (all from the Adolescent Brain & Cognitive Development Study) were 11.9 ± 0.7 years of age and the 4 groups were matched for sex, self-ratings of English, nonverbal reasoning, and combined household income. Structural magnetic resonance images were analyzed using CAT12 and all four groups were entered into a factorial analysis. RESULTS/ANTICIPATED RESULTS: Surprisingly, the main effect of Reading Ability did not reveal any regions where RD manifested less CT than Controls (raising the possibility that the findings from the only two prior reports were due to small samples). The main effect of Language Background revealed less CT in bilinguals in bilateral perisylvian regions (inferior frontal gyri, superior temporal gyri, and left Heschl's gyrus) consistent with prior reports. There was no interaction of Reading Ability by Language Background. Taken together, we found no differences in CT in those with RD relative to Typical readers and no evidence that the dual language experience affected this result in any way. DISCUSSION/SIGNIFICANCE: The lack of interaction between Reading Ability and Language Background indicates that a dual-language experience does not affect CT differently in those with RD and reduces concerns that RD in those who are bilingual needs to be given separate consideration in studies of CT neuroanatomy.
509 AMG487, A CXCR3 Antagonist, changes the Inflammatory Milieu in Familial Hemophagocytic Lymphohistiocytosis (FHL) Hepatitis
Authors: Diamond Tamir , Lau Michelle M. , Chu Niansheng , Behrens Edward M.
Published: April 2023

OBJECTIVES/GOALS: Familial Hemophagocytic Lymphohistiocytosis (FHL) is a systemic inflammatory disease, causing acute liver failure (ALF). Elevated Interferon gamma (IFN-γ) results in increased hepatic transcription of the chemokines CXCL9 and CXCL10. Inhibition of their receptor CXCR3 may reduce leukocyte recruitment and ameliorate hepatitis. METHODS/STUDY POPULATION: To determine the functional role of the IFN-γ-induced ligands, CXCL9 and CXCL10, in hepatic leukocyte recruitment via CXCR3 we used a prf-/- mouse infected with Lymphocytic Choriomeningitis Virus (LCMV) in our well-established model mimicking human FHL. We used AMG487, a small molecule CXCR3 antagonist, while maintaining intact IFN-γ signaling. Mice were sacrificed 10 days after infection when mice developed features of FHL: cytopenias, organomegaly, elevated serum ferritin and sCD25, and hepatic inflammation. Hepatic inflammation was characterized using flow cytometry, liver histology and noninvasive markers of hepatitis (ALT, liver size). RESULTS/ANTICIPATED RESULTS: AMG487 did not ameliorate the overall disease phenotype with mice developing similar FHL characteristics compared to control, including weight loss, elevation of ALT and sIL-2r as well as degree of thrombocytopenia and anemia. There was significant reduction of recruitment of CXCR3+CD4+ T cells and B cells in mice treated with AMG487. This indicates the importance of CXCR3 receptor in humoral response in FHL hepatitis. In addition, treatment with AMG487 resulted in reduction of CXCR3 expression in hepatic inflammatory monocyte (iMonos) measured by mean fluorescence intensity (MFI). DISCUSSION/SIGNIFICANCE: This is the first pre-clinical experience using AMG487, a small molecule CXCR3 antagonist, to treat FHL hepatitis. AMG487 changed the hepatic inflammatory milieu, reducing CD4 T-cell and B-cell recruitment, as well as CXCR3 expression on iMonos. However, it did not ameliorate FHL hepatitis and other therapeutic approaches should be pursued.
512 Odorant exposure decreases mortality in a Dravet Syndrome mouse model
Authors: Nobis William , Huffman Ragan , Mitchell Alyssa , Hannalla Martina
Published: April 2023

OBJECTIVES/GOALS: Our goal was to explore the actions of odorants on mortality and seizures in a DS mouse model (scn1a+/-), which have spontaneous seizures and high rate of SUDEP. We hypothesize that odorants that have actions on olfactory->;extended amygdala pathways will decrease SUDEP, potentially through attenuation of neuronal activation in the extended amygdala. METHODS/STUDY POPULATION: Dravet syndrome mice (heterozygous scn1a+/- ) were exposed for at least eight hours a day to either 2-phenylethanol (2PE, rose odor ), lemon extract, or vehicle odorant in group housed cages. This was repeated daily for 15 days starting at postnatal day 20/21. Mortality in each group was recorded. A subset of 2PE-exposed animals had an extended washout period following odorant exposure to continue to determine the long-term effect of odorant exposure on mortality. RESULTS/ANTICIPATED RESULTS: Our preliminary results show a strong trend for decreased mortality in the 2PE-exposed group (16.1% mortality (n=31) vs 38.5% mortality in vehicle control (n=26), p=0.06, Barnard’s test). Survival analyses show similar results (p=0.056 Kaplan-Meier curve, p=0.046 when removing those animals that died before completing day one of exposure). The lemon scent-exposed animals had a non-significant increase in mortality compared to controls from our preliminary experiments (50% mortality, n=8). Overall, these results suggest that mortality effect is dependent on specific odorants and that this effect is transient. DISCUSSION/SIGNIFICANCE: Our preliminary data support that odorant exposure can decrease mortality in a Dravet Syndrome mouse model, suggesting that more work to determine the mechanism of action and circuitry involved may illuminate new targets and therapies for preventing SUDEP in epilepsy patients.
A physics-based domain adaptation framework for modeling and forecasting building energy systems
Authors: Xuereb Conti Zack , Choudhary Ruchi , Magri Luca
Published: April 2023

State-of-the-art machine-learning-based models are a popular choice for modeling and forecasting energy behavior in buildings because given enough data, they are good at finding spatiotemporal patterns and structures even in scenarios where the complexity prohibits analytical descriptions. However, their architecture typically does not hold physical correspondence to mechanistic structures linked with governing physical phenomena. As a result, their ability to successfully generalize for unobserved timesteps depends on the representativeness of the dynamics underlying the observed system in the data, which is difficult to guarantee in real-world engineering problems such as control and energy management in digital twins. In response, we present a framework that combines lumped-parameter models in the form of linear time-invariant (LTI) state-space models (SSMs) with unsupervised reduced-order modeling in a subspace-based domain adaptation (SDA) approach, which is a type of transfer-learning (TL) technique. Traditionally, SDA is adopted for exploiting labeled data from one domain to predict in a different but related target domain for which labeled data is limited. We introduced a novel SDA approach where instead of labeled data, we leverage the geometric structure of the LTI SSM governed by well-known heat transfer ordinary differential equations to forecast for unobserved timesteps beyond available measurement data by geometrically aligning the physics-derived and data-derived embedded subspaces closer together. In this initial exploration, we evaluate the physics-based SDA framework on a demonstrative heat conduction scenario by varying the thermophysical properties of the source and target systems to demonstrate the transferability of mechanistic models from physics to observed measurement data.
Active screening and patient-placement and cohort-placement strategies to decrease carbapenem-resistant gram-negative bacilli colonization and infection in pediatric patients: A 5-year retrospective observational study in China
Authors: Yin LJ , He LY , Lu GP , Cao Y , Wang LS , Zhai XW , Wang CQ
Published: April 2023

Carbapenem-resistant gram-negative bacilli (CR-GNB) colonization screening was initiated across high-risk departments (PICU, NICU, neonatal wards, and hematology departments) in January 2017, and several CR-GNB cohort and patient-placement strategies were introduced throughout the hospital in January 2018. The colonization and infection rates decreased to varying degrees from 2017 to 2021.
Bacterial transfer and biofilm formation in needleless connectors in a clinically simulated in vitro catheter model
Authors: Ryder Marcia , deLancey-Pulcini Elinor , Parker Albert E. , James Garth A.
Published: April 2023

Objective:

Although needleless connectors (NCs) are widely used in clinical practice, they carry significant risk of bloodstream infection (BSI). In this study, we quantified differences in bacterial transfer and biofilm formation between various NCs.

Design:

Prospective, clinically simulated in vitro experimental study.

Methods:

We tested 20 NCs in a 5-day clinical simulation of Staphylococcus aureus inoculations onto NC septum surfaces, which were then flushed with saline and cultured for bacterial transfer. Biofilm formation was measured through destructive sampling of the connector-catheter system. Moreover, 8 NC design factors were evaluated for their influence on bacterial transfer and biofilm formation. This study was designed without a disinfection protocol to ascertain the intrinsic risk of each NC.

Results:

Clave Neutron and MicroClave had the lowest overall mean log density of bacteria in the flush compared to other NCs (P < .05), except there were no statistically significant differences between Clave Neutron, Microclave, SafeTouch, and SafeAccess (P ≥ .05). The amount of biofilm in the NC was positively associated with bacteria in the flush (P < .0005). Among 8 design factors, flow path was most important, with the internal cannula associated with a statistically significant 1 log reduction (LR) in bacteria in the flush (R2 = 49%) and 0.5–2 LR in the connector (R2 = 34%). All factors together best explained bacteria in the flush (R2 = 65%) and biofilm in the connector (R2 = 48%).

Conclusions:

Bacterial transfer and biofilm formation in the connector-catheter system varied statistically significantly between the 20 NCs, suggesting that NC choice can lower the risk of developing catheter-related BSIs.
Discontinuation of admission screening for coronavirus disease 2019 (COVID-19) and the impact on in-hospital clusters of COVID-19: Experience at a tertiary-care center
Authors: Honda Hitoshi , Nakagawa Masataka , Nisida Rie , Shintani Chiyo , Hamagishi Manami , Uehara Yuki , Iwata Mitsunaga , Doi Yohei
Published: April 2023

We evaluated the impact of discontinuing universal preadmission screening for severe acute respiratory coronavirus virus 2 (SARS-CoV-2) on the occurrence of nosocomial clusters of coronavirus disease 2019 (COVID-19) during the SARS-CoV-2 o (omicron) variant period. No increasing trend in nosocomial clusters was observed during community-based surges before and after discontinuation. This finding supports the safety of the practice change.
Secondary flows in turbulent boundary layers developing over truncated cone surfaces
Authors: Kaminaris Ioannis K. , Balaras Elias , Schultz Michael P. , Volino Ralph J.
Published: April 2023

The evolution of a turbulent boundary layer over truncated cones in staggered and random configurations is investigated using direct numerical simulations. Three random planform densities and one staggered, which closely match the configuration in the recent experiment by Womack et al. (J. Fluid Mech., vol. 933, 2022, A38), are considered. The experiment is also used to validate the present computations which are found to reproduce the flow measurements very accurately. Spanwise heterogeneity in the mean streamwise velocity in the form of high and low momentum pathways was detected in both the staggered and random arrangements, indicating the presence of secondary flows in the cross-stream plane. The momentum pathways found in the staggered arrangements are much weaker and confined near the roughness crest, while in the random arrangements they approach the edge of the boundary layer. We found a clear correlation of the momentum pathways with the leading edge of the roughness, while the correlation to the local topography was very weak for all the cases studied. Vorticity transport analysis is also employed to explore the nature of the secondary flows formed, highlighting the strong presence of vortex stretching near the leading edge of the roughness. The impact of the momentum pathways on the flow statistics, especially in the random arrangements, is demonstrated by the breakdown of outer-layer similarity.