bluuebunny/crossref_metadata_2025 huggingface dataset:
429 Elevated Phosphate Levels Induce Lung Inflammation and Exacerbate Pulmonary Fibrosis
Authors: Bollenbecker Seth , Easter Molly , Vang Shia , Hirsch Meghan , Barnes Jarrod , Faul Christian , Krick Stefanie
Published: April 2022

OBJECTIVES/GOALS: Using a cell culture model, we will determine the effects of phosphate on primary lung cell cultures and use our results to delineate a pathway through which these changes are carried out. Using animal models, we will determine the effects of phosphate on inflammatory and fibrotic lung injury, both in the presence and absence of CKD. METHODS/STUDY POPULATION: For our in vitro experiments, human lung fibroblasts were treated with concentrations of 1 to 5 mM sodium phosphate and FGFR inhibitors. Expression levels of interleukin (IL)-1beta, IL-6, and IL-8 were analyzed by qPCR and secretion of these cytokines was measured by ELISA. Phosphorylation of PLCy and ERK was measured by western blot. Using an in vivo approach, we placed C57Bl/6 mice on a high phosphate (3%) diet to elevate serum phosphate levels in the absence of kidney injury and administered bleomycin via oropharyngeal aspiration to generate an acute inflammatory response. Serum FGF23 levels were measured by ELISA and serum analysis for phosphate and renal function were obtained. Furthermore, expression of FGF23 pathway and inflammatory markers were analyzed in murine lung tissue using qPCR and western blotting. RESULTS/ANTICIPATED RESULTS: Augmented phosphate concentrations led to increased cytokine expression and secretion from human lung fibroblasts as well as a concomitant increase in PLCy and ERK phosphorylation. Inhibition of FGFR1 reversed the effects of phosphate on the inflammatory cytokines and PLCy/ERK phosphorylation. Serum FGF23 levels were significantly upregulated in mice on a high phosphate diet and further increased in mice subjected to a high phosphate diet with exposure to bleomycin. Both serum phosphate and creatinine levels were significantly elevated as well. Additionally, high phosphate and bleomycin increased local FGF23 expression in murine lung tissue, when compared to controls or each stimulus alone. DISCUSSION/SIGNIFICANCE: Phosphate has a significant impact on inflammation and fibrosis in the lung, indicating that the existence of pulmo-renal crosstalk exaggerates pulmonary injury and that there are biological pathways that may be targeted therapeutically to mediate these effects. These results could have a substantial impact on the quality of life for CKD patients.
430 Time restricted feeding in diet induced obesity mouse model reduces aortic stiffness and inflammatory T cells
Authors: Edell Claudia , Pati Paramita , Colson Jackson , Molina Patrick , Pollock David M , Pollock Jennifer S
Published: April 2022

OBJECTIVES/GOALS: Time restricted feeding (TRF) in diet induced obesity (DIO) has several health benefits, including improved metabolic rhythms and inflammation. Our lab has shown that TRF in DIO significantly reduces renal and aortic damage. The main goal of our research is to understand how TRF impacts aortic function, organ damage, and T cell activation in DIO. METHODS/STUDY POPULATION: We will use a 20-week DIO model, where mice will be on 20 weeks of normal fat diet (ND) or high fat diet (HFD). During weeks 18-20, mice will go through TRF intervention
where food is restricted to the 12-hour active period or continue ad libitum feeding. At the end of the 2-week TRF intervention or continued ad libitum feeding, aortic stiffness will be measured via pulse wave velocity measurements. We will also collect kidney, aorta, and small intestine at the end of the 20-week protocol for flow cytometric analysis of tissue T cell activation as well as histological assessments. This will allow us to determine the relationship with organ damage, organ function, and the T cell response. We will also analyze tissue and circulating levels of inflammatory T cell-derived cytokines such as interleukin-17A (IL-17A) via ELISA. RESULTS/ANTICIPATED RESULTS: DIO mice showed significantly increased aortic stiffness (measured by pulse wave velocity) compared to mice on ND. Interestingly, TRF intervention in DIO mice reduced aortic stiffness compared to DIO ad libitum. Histological assessments also showed that TRF abolished aortic and kidney fibrosis suggesting a role for the timing of feeding in regulating aortic function and organ damage from chronic HFD. We have several ongoing experiments to determine the T cell response with TRF in DIO mice. We predict that TRF in DIO mice will significantly decrease inflammatory T cells and reduce cytokine abundance in target organs. DISCUSSION/SIGNIFICANCE: Our lab has shown that TRF reduces aortic thickness and aortic and kidney fibrosis, but the driving mechanisms are unknown. We propose that TRF reduces T cell activation in DIO mice leading to reduced organ damage. Our work will provide insight on how TRF in DIO regulates the T cell response and may improve inflammation in the kidney and aorta.
433 Is miR let-7c protective against Acute Chest Syndrome in Sickle Cell Disease?
Authors: Fan James , Gemel Joanna , Lang Theodore , Beyer Eric , Lapping-Carr Gabrielle
Published: April 2022

OBJECTIVES/GOALS: We have shown that small extracellular vesicles (exosomes) isolated from patients with a history of ACS disrupt the endothelium in vitro. Sequencing of miRNA contents of these vesicles suggested that miR let-7c was differentially expressed. The current study was designed to determine the relationship between miR let-7c levels and ACS. METHODS/STUDY POPULATION: We identified 16 subjects from the SCD Lungomics biobank at the University of Chicago Comer and La Rabida Childrens Hospitals who had samples obtained at baseline. Among them, 9 had a history of ACS (ACS(+)) and 7 did not (ACS(-)). For all subjects, we reviewed clinical data relevant to their SCD and laboratory data (including hemoglobin, absolute reticulocyte count, white blood cell count) obtained at the same time as the baseline samples. RNA was isolated from the plasma and miR let7c was quantified using quantitative RT-PCR. RESULTS/ANTICIPATED RESULTS: Subjects were similar clinically, except that those with a history of ACS were more likely to be on hydroxyurea (p<0.05) and to have obstructive sleep apnea (p<0.05). Hematologic laboratory values were similar irrespective of ACS history. The mean miR let-7c level was 2-fold less in subjects with a history of ACS than in those who had never had ACS (p<0.05). A plasma miR let-7c level < 1 (normalized to the control subjects) had a positive predictive value of 0.78 for history of ACS and a sensitivity of 78% and specificity of 71%. Among subjects with a history of ACS, the let7c levels did not correlate with time since the last ACS event. However, among subjects who developed ACS following the baseline samples, higher miR let-7c levels correlated with increased length of time to next ACS event (R=0.8). DISCUSSION/SIGNIFICANCE: Our results in a group of subjects with SCD show that plasma miR let-7c levels are decreased in subjects with a history of ACS. They suggest that miR let-7c may be protective against development of ACS and that measurement of its levels could be a useful biomarker to assess or predict risk for this complication of SCD.
436 Examining the Role of Obesity and Leptin Signaling in Triple Negative Breast Cancer
Authors: Brock Courtney , Wright Maryl K. , Nguyen Khoa , Hebert Katherine , Alzoubi Madlin , Cheng Thomas , Collins-Burow Bridgette M. , Burow Matthew E.
Published: April 2022

OBJECTIVES/GOALS: In triple negative breast cancer (TNBC), obesity is associated with poor outcomes. Adipose stem cells (ASCs) from obese patients (obASCs) secrete higher levels of adipokines compared to ASCs from lean individuals. Leptin, one of these adipokines, has been implicated in many cancers. This study seeks to examine the role of leptin signaling in TNBC. METHODS/STUDY POPULATION: Previous work in conjunction with a collaborating lab has shown that leptin signaling promotes metastasis and increased expression of epithelial-mesenchymal transition (EMT) markers in triple negative breast cancer cell lines. This project expands upon this work through using both patient-derived cell lines and and patient-derived xenografts (PDX), and examines the role of leptin signaling both in vitro and in vivo. To determine the effects of obesity upon a PDX model of TNBC, a high fat diet was used to induce obesity in vivo. A pharmacological inhibitor of the leptin receptor was used to test the requirement for leptin signaling both in vivo and in vitro. RESULTS/ANTICIPATED RESULTS: Exposure to conditioned media harvested from obASCs increased the percentage of TNBC cells that expressed cancer stem cell markers, whereas exposure to an inhibitor of the leptin receptor decreased the percentage of cells with cancer stem cell markers. PDX tumors implanted into mice with diet-induced obesity had an increased volume compared to tumors implanted into lean controls. Further analysis will be conducted on metastasis, circulating tumor cells, and survival in both lean and obese mice. DISCUSSION/SIGNIFICANCE: Understanding the complex signaling events in the obese tumor microenvironment is essential, as these molecular differences may contribute to different outcomes for obese and lean individuals with triple negative breast cancer. Therefore, study of the crosstalk between obASCs and TNBC cells is critical.
438 “How do you define Resilience?” Examining the Psychological Resilience of Black Adults Living with Sickle Cell Disease (SCD)
Authors: Conn Corinne April Iolanda , Creary Melissa , Bonham Vence
Published: April 2022

OBJECTIVES/GOALS: This study examines psychological resilience in adults living with Sickle Cell Disease (SCD) in the U.S. aiming to explore how psychological resilience is defined by this community. METHODS/STUDY POPULATION: Participants were recruited between 2014 – 2018, from across the U.S. as part of an ongoing cross-sectional study: Insights into Microbiome and Environmental Contributions to Sickle Cell Disease and Leg Ulcers Study (INSIGHTS). Inclusion criteria included age of 18 or older, with a clinical history of SCD, and were interviewed if they completed the Brief Resilience Scale (BRS) as part of INSIGHTS. 150 study participants were separated by their BRS scores into “High” and “Low” scoring quartiles. 30 participants were randomly selected, 15 from the lowest quartile and 15 from the highest. All participants completed the Connor Davidson Resilience (CD) measure at the end of their qualitative interview. All identified as Black with an average age of 42.5 (13 F, 17 M). RESULTS/ANTICIPATED RESULTS: Three main concepts emerged within both groups in response to the question “How do you define resilience?” (a) not giving up (b) how one deals with challenges and (c) moving forward. DISCUSSION/SIGNIFICANCE: Our analysis shows that the BRS may not be a precise or accurate indicator of the resilience of adults living with SCD. Therefore, it remains to be seen if these measures are descriptive of these individuals true psychological or physiological state as they have not been used in this community until now.
439 The effect of non-invasive transcutaneous auricular vagus nerve stimulation (taVNS) on hypoxic-ischemic injury in newborn rats
Authors: Wiley Melanie Gail , Sims-Robinson Catrina , Boger Heather A. , Jenkins Dorothea D. , George Mark S. , Johnson Ralph H.
Published: April 2022

OBJECTIVES/GOALS: Neonatal hypoxic-ischemic encephalopathy (HIE) is an acute neurologic syndrome where decreased blood flow and oxygen to the brain causes acute and chronic brain dysfunction. The only proven neuroprotective intervention for HIE is hypothermia treatment started within 6 hours of birth and 50% of survivors have long-term deficits. METHODS/STUDY POPULATION: Pre-clinical adult stroke studies demonstrated that vagus nerve stimulation (VNS) has anti-inflammatory effects and attenuates brain damage. Transcutaneous auricular VNS (taVNS) is safe and feasible in infants and may improve the motor skill of bottle feeding. We hypothesize that a combined hypothermia-taVNS treatment shortly after HIE birth will have neuroprotective effects, improve motor function, attenuate infarct volume inflammation compared to hypothermia alone. The HIE model includes ligation of the right common carotid artery in postnatal day 7 (P7) rats followed by 90min hypoxia (8% oxygen) and 2hr hypothermia. taVNS or sham taVNS was administered using a bipolar electrode placed on the auricular concha region for 30min, [30sec trains, 0.5msec duration, 20Hz frequency, followed by 4.5min breaks] RESULTS/ANTICIPATED RESULTS: Experimental groups include +HIE/+taVNS, +HIE/-taVNS, and -HIE/-taVNS. To assess motor function, grasping reflex and forelimb grip strength tasks were assessed prior to surgery through P10. Infarct volume was assessed at 72h after injury by staining coronal sections with cresyl-violet. Thirty-four rat pups underwent surgery with an 8.82% mortality rate. taVNS was well tolerated by the P7 rats when delivered below perceptual threshold (0.4-1.1mA). There was no difference in elementary motor function or infarct volume between any group. DISCUSSION/SIGNIFICANCE: Future studies will include 2.5hr hypoxia for a more severe brain injury and a -HIE/+taVNS control group. These initial pre-clinical studies in neonates are important in determining whether taVNS may translate as a treatment to improve outcomes after neonatal HIE.
443 COVID19 disease severity influences the expression of markers of durability in memory B cells
Authors: Reyes Raphael , Clarke Kathleen , Cantwell Angelene M. , Catano Gabriel , Tragus Robin E. , Patterson Thomas F. , Bol Sebastiaan , Bunnik Evelien M
Published: April 2022

OBJECTIVES/GOALS: Studies have shown that SARS-CoV-2 specific memory B cells can be maintained at least a year after exposure. However, reports show an altered B cell response during infection in severe COVID-19 cases. This study aims to describe the B cell response during COVID-19 convalescence with a focus on signatures that contribute to durable and robust immunity. METHODS/STUDY POPULATION: Our study cohort consisted of individuals who had recovered from non-severe (hospitalized) or severe (hospitalized and requiring invasive mechanical ventilation) COVID-19. In our comparative analysis, samples from both groups were carefully matched to fall within 4-5 weeks post-symptom onset. We also performed a longitudinal analysis of non-severe patients with sampling ending 5 months post-symptom onset. Using high parameter flow cytometry, we characterized the phenotype of memory B cells using 19 distinct cell markers and fluorescently labeled probes to identify B cells reactive with SARS-CoV-2 spike and receptor-binding domain protein. Additionally, serum collected from individuals was used to quantify antibody titers. RESULTS/ANTICIPATED RESULTS: The frequency of spike-specific B cells and serum antibody titers were similar between severe and non-severe groups. However, we observed that individuals recovered from severe COVID-19 have a significantly reduced frequency of spike specific IgG+ memory B cells expressing Tbet and FcRL5 (markers associated with long lived immunity). In the non-severe patients, we observed IgG+Tbet+ B cells targeting the spike protein peak at 2-3 weeks post-symptom onset, decrease by almost fifty percent 4-5 weeks post-symptom onset, and return to baseline 5 months post-symptom onset. Our study also validated previous findings of a short-lived primary response of IgM+ B cells targeting the spike protein. DISCUSSION/SIGNIFICANCE: Our findings highlight potential implications for long-term immunity against re-infection or severity of the resulting disease in patients with severe COVID-19. Further investigation will be necessary to determine whether the maintenance of immunological protection is hindered in patients who overcame severe COVID-19.
444 Post-translational role of RNA modifications in sRNA chaperone Hfq
Authors: Nurse Jalisa , Joseph Aubee , Thompson Karl M
Published: April 2022

OBJECTIVES/GOALS: The goal of this study is to determine the role of the tRNA modifications in the translation of Hfq. Hfq is an RNA chaperone that acts as a co-factor for the action of the largest class of small RNAs in E. coli. RNA modifications have been known to play critical roles in the translational fidelity of many cellular proteins in bacteria. METHODS/STUDY POPULATION: In this study, we used an hfq-lacZ translation fusion to screen several RNA modification mutant genes to uncover additional RNA modifications that may play a role in Hfq translation. We measured hfq-lacZ activity in genetic backgrounds mutated for several additional RNA modification enzymes previously untested for Hfq effects. RESULTS/ANTICIPATED RESULTS: We identified 5 RNA modification genes that were defective for hfq-lacZ fusion activity, and we subsequently performed western blot analysis on the Hfq protein in the absence of these modification mutant genes to determine the effect of these mutants more directly on Hfq protein levels. We identified 2 out of these 5 RNA modification mutants that also affect Hfq protein levels. DISCUSSION/SIGNIFICANCE: Since Hfq is critically important for small RNA function is a wide range of bacteria, it is possible tRNA modifications regulate Hfq expression in other bacteria. These processes, when further investigated, could provide us with the basic information to develop new antibiotics needed to address emerging antibiotic resistance.
445 Inhibition of GPR30 Reveals Putative Genes Involved in the Pathogenesis of Inflammatory Breast Cancer
Authors: Carrion-Laureano Melanie , Acosta-Montalvo Melvyn , Bittman-Soto Xavier , Peterson-Peguero Esther , Pérez-Santiago Josué
Published: April 2022

OBJECTIVES/GOALS: Inflammatory Breast Cancer (IBC) is the most aggressive form of breast cancer and does not have targeted therapy. GPR30, a 7-transmembrane estrogen receptor, may play a role in regulating cell growth and proliferation of cancerous cells. Here, we evaluated changes in gene expression while inhibiting GPR30 to determine putative targets to treat IBC. METHODS/STUDY POPULATION: IBC cell lines (SUM149PT) were cultured in medium with serum stripped from growth factor and hormones for 48 hours. Cells were then exposed to either G15 (GPR30 inhibitor) at a concentration of 1ÂµM or ETOH (vehicle negative control) 3 hours in triplicates. After exposure, total RNA was extracted using the Qiagen RNAeasy Mini kit and RNA was sequenced using the Illumina NextSeq (2 X 75bp). The higher-quality reads were aligned, annotated, and quantified to the human genome (HG38) using STAR and RSEM softwares. Gene expression analysis was performed in R statistical software (packages tximport and DESeq2). Functional and enrichment analyses were performed using Metascape and STRING database, respectively. RESULTS/ANTICIPATED RESULTS: There were 656 significantly expressed genes (p < 0.05) between groups (G15 vs. ETOH). The top 5 significant genes include: SMIM7, FANCG, ARID1A, MAML2, and ATF3. Significantly impacted biological processes and pathways include: electron transport chain, mitotic cell cycle process, microtubule cytoskeleton organization, cellular component morphogenesis and DNA-dependent DNA replication (adj p < 0.05). Additionally, physical and functional interaction networks showed 3 major clusters (â‰¥ 12 genes), which contained several gene hubs including BRCA1, BRCA2, FOS (proto-oncogene), PLK1 and PAK1 (both serine/threonine-protein kinases), among others. Interestingly, the network analysis showed the previously known interaction between FANCG and BRCA2, which were both dysregulated by GPR30 inhibition. DISCUSSION/SIGNIFICANCE: Through gene expression, functional and enrichment analyses we found several targets genes that could be associated with the pathogenesis of IBC. Validation of candidates genes (qRT-PCR and Western blot), and functional assays (cell proliferation, motility, and invasion) will be performed to understand the potential of these genes in treating IBC.
448 Isolation and identification of bioprospects capable of metabolizing 17-beta-estradiol and 17-alpha-ethinylestradiol using metagenomics and culture-dependent techniques in Puerto Rico
Authors: Ríos-Velázquez Carlos , Cabrera-Ruiz Melissa M. , Delgado-Serrano Kristal M. , Morales-Lajara Hailey N.
Published: April 2022

OBJECTIVES/GOALS: This research project aims to isolate and identify bioprospects capable of metabolizing estrogen using culture-dependent, culture independent methods and the identification of the gene/genes responsible for the metabolization of estrogen by the bioprospects. METHODS/STUDY POPULATION: For the culture dependent technique, samples were collected from the water treatment plant in MayagÃ¼ez, cultivated on TSA medium and selected specific and diverse colonies were patched on M-9NC (no carbon sources), M-9-glucose (M9G) and M-9-hormone mixture (M9H: 17-beta-Estradiol and 17-alpha-Ethynylestradiol). After the 48hrs incubation at 25 and 37 Celsius, growth was scored on the different media, to choose those potential bioprospects that use the hormones as the sole carbon source. For the culture independent approach, metagenomic clones from libraries generated from the Guajatacas water reservoir and Cabo Rojos Saltern microbial mats were cultivated on the appropriate M9NC, M-9G and M-9H, supplemented with Leucine and Thiamine for 48 hours at 37 Celsius. RESULTS/ANTICIPATED RESULTS: The culture-dependent approach showed a total of 6 potential bioprospects capable of growing on hormones as a sole carbon source on M9H at 25 and 37 Celsius and no-growth on the M-9 media used. Currently, we are in the process of determining the identities of the cultivable isolates genetically, any preference on hormone metabolization, monitoring a larger number of metagenomic clones and optimizing the selection conditions. DISCUSSION/SIGNIFICANCE: The identification of bioprospects capable of metabolizing hormones could potentially lead to the generation of new water treatments that could help remove unwanted chemicals, such as high concentrations of estrogens.
453 Efficacy of the Insulin Infusion Calculator Protocol in the Optimization of Perioperative Blood Glucose Levels in the Cardiac Surgical Patients at UAB
Authors: Bode John , Stewart Hayden , Ballenger Angela , Boutwell Gary , Zaky Ahmed
Published: April 2022

OBJECTIVES/GOALS: Insulin dosing is crucial for regulating blood glucose in cardiac surgery patients, yet it requires the use of cumbersome insulin dosing charts. To streamline this process, an electronic insulin calculator (IC) is trialed in the cardiac operating rooms and intensive care unit. This study compares glycemic control prior to and after institution of IC. METHODS/STUDY POPULATION: Using the EHR, a retrospective population of 3,164 cardiac surgery patient charts from 12/19/19 - 11/06/20 were obtained. Baseline, intra-operative, and ICU blood glucose values were obtained from each patients admission. Using this data, a baseline level of glycemic control throughout a cardiac surgery patients stay was established. A preliminary cohort of 244 patients were then chosen to be managed with the new IC. Baseline, intra-operative, and ICU blood glucose values were obtained in this group for comparison. Paired t tests were used to compare the IC group of patients to the retrospective group of patients. Additional subgroup analysis was performed to assess IC efficacy for on pump vs off pump cardiac surgeries. RESULTS/ANTICIPATED RESULTS: The 244 patients managed with the IC showed significantly reduced average blood glucose values during their time in the ICU compared to those previously not managed with the IC (185 mg% vs 153 mg%, p= 0.02). Additionally, a trend towards a reduction in last operating room blood glucose level was also noted. Lastly, average blood glucose levels were significantly reduced for patients undergoing on pump cardiac surgeries compared to off pump surgeries (157 mg% vs 149 mg%, p = 0.03). DISCUSSION/SIGNIFICANCE: Preliminary results suggest IIC to be associated with better intra and postoperative blood glucose control. More data is being collected to test its association with outcomes.
457 Semaphorin7a expression in breast cancer promotes susceptibility to immune checkpoint blockade
Authors: Elder Alan , Stoller Alexander , Kines Kelsey , Lyons Traci
Published: April 2022

OBJECTIVES/GOALS: The goal of this study is to decipher the mechanisms by which breast cancers evade the immune system to facilitate metastasis through the lymphatic vasculature. We believe this is caused, in part, by semaphorin 7A (SEMA7A) as its expression is associated with increased lymphatic vessels, tumor-associated macrophages, and metastasis in breast cancer. METHODS/STUDY POPULATION: We have observed that breast cancer cells, lymphatics, and macrophages exhibit SEMA7A-dependent expression of PD-L1 in vitro, which may make SEMA7A+ tumors more sensitive to anti-PD-1/PD-L1 immunotherapies. Therefore, we utilized multiple orthotopic, immunocompetent mouse models to determine if SEMA7A expression makes tumors more susceptible to immune checkpoint blockade. E0771 and 66cl4 mouse mammary carcinoma cells were injected orthotopically into the #4 mammary fat pads. Once tumors reached 50-100 mm^3 mice were injected intraperitoneally with 250ug of anti-PD-1, anti-PD-L1, or IgG control every third day. Tumors were measured with calipers every other day and harvested for flow cytometry to determine the effect of immune checkpoint blockade on the TME in SEMA7A+ tumors. RESULTS/ANTICIPATED RESULTS: We reveal that growth of SEMA7A overexpressing (OE) tumors, but not controls, was significantly slowed with both anti-PD-1 and anti-PD-L1 treatments. Flow cytometric analysis of cells from the TME revealed increased LECs, TAMs, and PoEMs in SEMA7A+ tumors, compared to controls – all populations had higher PD-L1 expression, which was decreased with both anti-PD-1 or anti-PD-L1. We also observed a decrease in PD-L1 expression on the tumor cells with treatment. Furthermore, LEC, TAM, and PoEM presence was decreased within the TME alongside increased presence of activated CD4 and CD8 T cells. Collectively, our results suggest that SEMA7A expression in breast cancers activates a major pathway associated with immune evasion and helps to establish a pro-tumor microenvironment. DISCUSSION/SIGNIFICANCE: SEMA7A expression establishes a pro-tumor microenvironment, which can be targeted with readily available FDA approved drugs such as immune checkpoint-based therapies. Since SEMA7A+ breast cancers have high rates of metastasis, more specific treatments for these patient populations should be explored.
458 Reproducibility of Maximum ATP Production Following Isotonic versus Isometric Exercise
Authors: Sanchez Noah Dionicio
Published: April 2022

OBJECTIVES/GOALS: This project aimed to investigate differences between isometric and isotonic systems for low and high frequency contractions. Intracellular changes in pH caused by the rate of exercise differ in their effect on PCr. The project assessed the reproducibility between these exercise methods. METHODS/STUDY POPULATION: Four subjects were studied (2F/2M, avg. age 27 years). 31P-MRS spectra were measured in the vastus lateralis (VL) using a 3T MRI scanner using a 31P/1H surface coil. In the isometric studies, the (supine) subjects legs were supported by a wedge pad and restrained by straps over the hips and knee with a resistance strap over one ankle. In the isometric studies, the (prone) subjects legs were fit into machine designed to induce isometric exercise. There were 4 exercise bouts that differed by exercise type and frequency and, consequently, duration. 31P-MR spectra were acquired from a 25mm slab of leg positioned parasagittal along the VL. Analyses were done using jMRUI and R. RESULTS/ANTICIPATED RESULTS: With an increase in sample size, we expect differences between recovery time constants, tau, of PCr between the different exercise frequency groups alluded to by other studies investigating recovery rates in bicep skeletal muscle. From the preliminary data, we obtained large limits of agreement of tau = 0.053 s-1 & tau =-0.074 s-1 from using Bland-Altman analysis. As we increase our sample size, we anticipate that we will see differences of PCr recovery rates between the exercise types as well. DISCUSSION/SIGNIFICANCE: Current studies that investigate the differences in PCr recovery rates between different metabolic states differ in the exercise methods employed on subjects. This study aims to standardize PCr recovery assessment in the VL for more interpretable and applicable assessments of metabolic function.
459 Parenting Stress and Drinking to Cope Association with Early Childhood Risk Behaviors
Authors: Bautista Tara , Sinha Rajita , Silverman Wendy , Jastreboff Ania
Published: April 2022

OBJECTIVES/GOALS: Parent stress and coping impacts reward and motivation circuits during child development which influence self-regulation. One well known maladaptive coping response is alcohol or drinking-to-cope (DTC). This study assessed differences in stress and child behaviors among DTC parents as compared to non-DTC parents. METHODS/STUDY POPULATION: Baseline data was used from parents of a 2-5-year-old who were screened for a larger study assessing a mindfulness-based parent stress reduction intervention to improve healthy choices for themselves and their families. The sample included 172 parent-child dyads, mean parent age was 34.4 (6.1) years old, 56.3% white, mean child age was 3.6 (1.2) years old, 52.3% male. Subjective stress was assessed using the Perceived Stress Scale (PSS), parent-specific stress was assessing using the Parenting Stress Index (PSI), DTC was assessed using the COPE inventory, and child behaviors were assessed using the Devereux Early Childhood Assessment. To investigate the differences in stress and child behaviors between DTC parents and non-DTC parents independent samples t-tests were conducted. RESULTS/ANTICIPATED RESULTS: DTC was significantly correlated with PSS (r= 0.23, p<.01), PSI (r= 0.26, p<.01), child self-control (r= -0.16, p= .03), child attention problems (r= 0.22, p<.01), and total behavioral concerns (r= 0.16, p=.04). After excluding those who do not drink alcohol at all, we found significantly higher perceived stress among DTC parents (M= 27.83, SD= 9.79) compared to non-DTC parents (M= 23.79, SD= 8.40), t(80)= 2.02, p= .02. For children, we found significantly higher aggression scores for children of DTC parents (M= 47.16, SD= 31.69) compared to children of non-DTC parents (M=35.83, SD=25.72), t(84)= 1.83, p= .04. And greater attention problems among children of DTC parents (M= 73.97, SD= 26.77) compared to children of non-DTC parents (M= 56.71, SD= 34.09), t(84)=2.63, p=.01. DISCUSSION/SIGNIFICANCE: Stress and DTC may contribute to negative behaviors in children. An intervention designed to decrease stress and increase adaptive coping mechanism in parents could benefit health child socioemotional and behavioral development. Future analyses will examine third variable effects in the relationship between stress, coping, and child behaviors.
462 Dysfunctional leukocyte mitochondrial metabolism is associated with immune paralysis in critically ill septic patients
Authors: Torres Lisa K , McConn Keith , Gomez-Escobar Luis , Cloonan Suzanne , Choi Augustine MK
Published: April 2022

OBJECTIVES/GOALS: The host immune response during sepsis is now recognized to have anti-inflammatory pathophysiology. We aim to determine whether mitochondrial dysfunction of leukocytes predicts which critically ill septic patients develop immune paralysis and to identify differences in cellular metabolites between patients with and without immune paralysis. METHODS/STUDY POPULATION: Critically ill septic and control adult patients were recruited from one of 6 ICUs in a single-center tertiary care academic hospital. After enrollment, peripheral blood mononuclear cells (PBMCs) were isolated from a tube of whole blood on day 0-1 after ICU admission. Flow cytometry to quantify monocyte HLA-DR was performed to determine whether patients were immune paralyzed or not. Mitochondrial functional assays of PBMCs were performed with inhibitors of the electron transport chain to assess for differences in oxidative phosphorylation and glycolysis utilization. Metabolic profiling of cell pellets was performed to evaluate for specific metabolites and pathways associated with immune paralyzed patients. RESULTS/ANTICIPATED RESULTS: A total of 101 patients were recruited, including 62 control and 39 septic patients. 81 patients had immune paralysis status available for analysis. 52% of all recruited subjects were immune paralyzed. Of these, 58% were controls and 75% were septic. Immune paralyzed septic and control patients showed features of reduced utilization of oxidative phosphorylation (ox phos) including reduced basal respiration, ATP production and maximal respiration compared with non-immune paralyzed septic and control patients. Immune paralyzed septic patients showed diminished glycolysis utilization compared with septic non-immune paralyzed patients. Finally, cellular kynurenine and quinolinate levels were low in both immune paralyzed control and septic patients compared with non-immune paralyzed patients. DISCUSSION/SIGNIFICANCE: The PBMCs of immune paralyzed septic patients show evidence of mitochondrial dysfunction, with reduced ox phos and glycolysis utilization. Low levels of kynurenine and quinolinate, metabolite precursors to NAD+, in immune paralyzed patients may signal key deficiencies and targetable therapeutic avenues for reversal of an immune paralyzed state.
467 Post-transcriptional regulation of the MiaA prenyl transferase by the small RNA CsrB in Escherichia coli (E. coli)
Authors: Aubee Joseph I , Adigun Alexandria , Branch Kirsten R , Conyer Ava C , Olusanya Olufolakemi , Williams Kinlyn , Thompson Karl M
Published: April 2022

OBJECTIVES/GOALS: MiaA is a highly conserved prenyl transferase that catalyzes synthesis of the i6A37 tRNA modification in E. coli. While transcriptional regulation of MiaA is well characterized, there is no information on the MiaA post-transcriptional regulation. The aim of this study is to characterize the post-transcriptional regulation of the MiaA gene in E. coli. METHODS/STUDY POPULATION: To characterize the post-transcriptional regulation of miaA, we executed a targeted genetic screen of an E. coli small RNA library on a miaA-lacZ translational reporter fusion strain to identify small RNAs (sRNAs) that modulate MiaA translation or transcription termination. We also measured MiaA mRNA levels and miaA-lacZ activity in the absence or over-expression of candidate sRNA regulators of MiaA. We also measured MiaA mRNA levels in the absence of RNaseE and PNPase, two enzymes involved in mRNA turnover. Finally, we measured the ability of purified recombinant CsrA to bind to the MiaA mRNA transcript in vitro. RESULTS/ANTICIPATED RESULTS: We identified the carbon sensing sRNA CsrB and its cognate protein interaction partner CsrA, as potential post-transcriptional regulators of MiaA. Over-expression of CsrB fully repressed miaA-lacZ activity and MiaA mRNA levels. The absence of CsrA resulted in a defective miaA-lacZ activity and a 10-fold decrease in MiaA mRNA levels. We also identified an increase in the MiaA mRNA half-life particularly in the absence of RNaseE. Our results demonstrate an additional layer of regulation for the miaA operon by the CsrA/CsrB protein-sRNA system. DISCUSSION/SIGNIFICANCE: MiaA is a highly conserved bacterial protein. Our data may represent phenomena in an array of bacteria that could be targeted by novel antibiotics. The human MiaA homologue, TRIT1, plays a role in mitochondrial disorders. We anticipate that information garnered from MiaA studies will elucidate TRIT1 function and its role in mitochondrial disorders.
472 Indiana CTSI Think Tank Program
Authors: Garcia Kara , Portonovo Padma , Garci Kara
Published: April 2022

OBJECTIVES/GOALS: The Think Tanks’ aim to assist faculty innovators along the path from discovery to commercialization; serving as a one-stop-shop where investigators can access advice, pilot funds, and direction toward other available resources within the Indiana CTSI. METHODS/STUDY POPULATION: Faculty receive guidance from a pool of advisors in the drug and device industry, as well as their respective university commercialization offices; who serve as an essential resource for a wide range of scientific, technical, clinical, business, and regulatory questions. Investigators submit a simple intake form to connect with a project facilitator for detailed guidance prior to formal meetings. Projects are guided from start to finish with robust tracking and milestone-based funding to help generate data for Investigational New Drug (IND) or Investigational Device Exemption (IDE) applications. Project tracking is conducted using REDCap, which is used for all investigator submissions, with internal processes also tracked in REDCap. RESULTS/ANTICIPATED RESULTS: From Feb-Aug 2021; the Think Tanks provided feedback to 15 drug projects and 6 medical device projects. This included 12 from Indiana University; 4 from Purdue University; and 5 from the University of Notre Dame. Interestingly; drug innovations were submitted primarily by tenure track faculty (100%) with a history of NIH/NSF funding (50%); while device innovations were submitted primarily by clinical faculty without a history of NIH/NSF funding (66%). Based on first-round feedback, a total of approximately $3;400 in pilot funds were provided. Efforts are underway to obtain survey-based feedback from all applicants to date; which will be used to inform future program modifications. DISCUSSION/SIGNIFICANCE: The budding “Think Tank” program provides faculty with a broad perspective of the entire drug and medical device development process, helping investigators understand the critical interplay development stages. Future work seeks to enhance faculty engagement in, and understanding of the commercialization process across Indiana CTSI institutions.
478 You liked it, but did you learn anything? A process for redesigning follow-up surveys in attempts to measure success beyond satisfaction
Authors: North Gemma , Osuna Shardae , Dezfuli Ghazal
Published: April 2022

OBJECTIVES/GOALS: Project objectives: Collect, analyze, redesign, and redeploy the follow-up surveys sent after services rendered or educational offerings attended to improve outcome measurement. Presentation objectives: Provide a process for others to optimize their assessment surveys. METHODS/STUDY POPULATION: A team of interdisciplinary experts from Evaluation and Improvement, Workforce Development, and Administration took a systematic and collaborative approach to optimizing service and educational offering assessment. The team collected all 35 existing surveys currently in use at the CTSA, developed a matrix table to organize findings, cross-analyzed/normed to recognize and reduce bias, engaged other staff and faculty at specific intervals to encourage buy-in, and responded to feedback. The team then updated both surveys and format, programmed revised surveys into a centralized platform, and provided instructions and training for implementation. RESULTS/ANTICIPATED RESULTS: This effort resulted in streamlining both the assessment process and the surveys used by the Southern California Clinical and Translational Science Institute (SC CTSI). Instead of using different versions of surveys or none at all, now SC CTSI commits to using the same follow-up survey for events and for educational offerings institute-wide, and agrees to store and access that data via a single platform, REDCap, allowing any member to see data in real time. This will allow SC CTSI to monitor and evaluate its short-term outcomes at an institutional level, and determine areas for improvement or best practices. Future plans include training on survey data interpretation for decision making. DISCUSSION/SIGNIFICANCE: Presenters will share lessons learned and considerations when embarking on streamlining assessment practices institute-wide, highlighting the importance of leveraging educational methodology to go beyond measuring satisfaction and into measuring learning. Presenters will share revised surveys.
493 The use of visual arts to teach complex ideas on non-binary sex/gender traits and identities.
Authors: de Jesús Maria Inés , Bravo Laura
Published: April 2022

OBJECTIVES/GOALS: Recent scientific discoveries show that human sex determination and differentiation is a spectrum of developmental processes and that sex/gender traits and identities not always fit binarity. This study aims to determine whether the visual arts can effectively transmit these complex scientific ideas on sex and gender variance. METHODS/STUDY POPULATION: A one-hour lecture for undergraduate students enrolled in a behavioral neuroendocrinology course included 17th century Spanish paintings and representative work from contemporary LGBTQIA+ artists in photography. Pre and post self-evaluation was conducted through five multiple choice questions. Chi Square test was employed for statistical analysis, which required elimination of no responses from item analysis. Statistical significance was defined as p value < 0.05. Three artistic images and two scientific images were included in the questions. Acquisition of content-specific knowledge on diverse body configurations, differences of sex development (DSD), transsexuality, histopathology of the gonad and neuroanatomy of the hypothalamus was evaluated. RESULTS/ANTICIPATED RESULTS: Twenty students completed the online pre- and post-test. We found that a 17th century Spanish painting was effective at making the distinction between transsexuality and intersexuality (p < 0.0005). Similarly, an artistic image that reinterprets the Vitruvius Man was effective for presenting the argument that diverse body configurations does not imply clinical pathology (p < 0.0001). Last, a scientific image showing the histopathology of an ovotestis was effective for introducing undergraduate students to diagnostic criteria for DSD (p < 0.02). No significant differences were found in the use of an artistic photograph depicting a young female to male transsexual individual nor in the use of a scientific image showing the neuroanatomical localization of the hypothalamus. DISCUSSION/SIGNIFICANCE: The learning of complex scientific concepts on human sex determination and differentiation can be affected by preconceived ideas, values, and attitudes towards sex and gender variance. The visual arts can provide a familiar ground of understanding between teachers and learners to transform such preconceptions.
498 Investing in Translational Science: Forging Critical Connections with Investment Professionals
Authors: Cook Samantha , Martin Bradley J. , Servoss Jonathan M.
Published: April 2022

OBJECTIVES/GOALS: Ask the Experts: A Biomedical Innovation Forum, presented by Fast Forward Medical Innovation (FFMI) at the University of Michigan, provided an opportunity to educate biomedical innovators on life science investment trends and technology assessment criteria. METHODS/STUDY POPULATION: FFMI, in partnership with the U.S. Economic Development Administration, recruited an expert group of panelists to be featured at this virtual event. These life science investment experts provided insight on the strategy, timing, and best method for innovators to engaged investors, the specifics of what investors look for in technologies and project teams, and expectations of investors and project teams after the investment is secured. The panel presentation was followed by a poster presentation highlighting projects from the FFMI Hub at the University of Michigan, allowing innovators to have an open and constructive conversation with experts and attendees. RESULTS/ANTICIPATED RESULTS: There was a total of 73 registrants including academic faculty, biomedical innovators, and life science investment professionals from 21 different academic institutions, private companies, and other organizations. 50 attended the panel presentation and poster session. Results (N=5) of an evaluation of the event revealed that 100% of the respondents strongly agreed or agreed that the event met their expectations, while 80% strongly agreed or agreed that they would recommend the event to a colleague. Feedback from poster presenters was also strong with presenters exclaiming they “enjoyed the panel discussion and getting one-on-one time with the panelists,” as well as “a lot of great advice was given by the experts” and “I really liked the poster presentation part in which I got feedback from the investors.” DISCUSSION/SIGNIFICANCE: The data demonstrates how accelerating technology mining activities, proactively seeking and strengthening external partnerships with investors, and scaling commercialization education programs can have a positive impact on the development and launch of biomedical innovations.
499 Visiting Scholars in Times of Pandemic
Authors: Moussatche Patricia , Meyer Jessica P. , Orasa Ellen , Tsao Peg , Asch Steven M.
Published: April 2022

OBJECTIVES/GOALS: The Virtual CTSA Visiting Scholars Program allows KL2 scholars to serve as visiting professors at host CTSA consortium institutions. This program facilitates connections with faculty outside scholars home institutions, as well as fostering collaboration among different CTSA hubs. Stanford aimed to incorporate participants into its KL2 program. METHODS/STUDY POPULATION: Stanford is one of 57 host institutions for the Virtual CTSA Visiting Scholars Program. The program includes a virtual meeting between each participant and a chosen faculty at the host institution. Program participants also give a virtual Grand Rounds lecture, open to the entire CTSA consortium. As well as encouraging Spectrum KL2 Scholars to participate in the virtual exchange, the Spectrum KL2 program incorporated the visiting scholars into weekly activities over the course of 3 months. Visiting scholars were invited to participate virtually in KL2 educational sessions, which provide career development training and mentoring. As meetings transitioned to in-person, KL2 scholars were allowed to attend virtually when needed. Hybrid in-person sessions were also conducive to participation by visiting scholars. RESULTS/ANTICIPATED RESULTS: Stanfords Spectrum KL2 program was virtual (Zoom) for academic year 2020-2021 and included 1:1 mentoring sessions, weekly career development seminars, and 1:1 peer virtual lunches to integrate the 9 junior faculty scholars. Visiting scholars joined the weekly Zoom meetings when their schedules allowed. KL2 faculty mentored visiting scholars for 3 months, exchanging ideas and forming collaborations. Each visiting participant was paired with a KL2 Scholar, who provided 1:1 peer mentorship. In addition, visiting scholars presented a work-in-progress seminar, to obtain feedback before the more formal Grand Rounds lecture. For academic year 2021-2022, Spectrum KL2 meetings include virtual and hybrid in-person sessions, allowing visiting scholars to join by Zoom during the 3-month virtual exchange program. DISCUSSION/SIGNIFICANCE: What set Stanfords Virtual CTSA Visiting Scholars Program apart was faculty engagement and mentorship provided to visiting participants. By incorporating visiting scholars virtually into the ongoing KL2 education program, participants could engage fully with mentors and scholars, even surpassing opportunities of pre-pandemic on-site visits.