bluuebunny/crossref_metadata_2025 huggingface dataset:
Putting Their Money Where Their Mouths Are: A Multidisciplinary Approach to Addressing Disparities in Health Research, Grant Funding, and Recruitment of Black Women to Breast Cancer Clinical Trials
Authors: Vogel Katherine , Okoruwa Oseme Precious , Ridley-Merriweather Katherine E. , Hoffman-Longtin Krista J. , Ashburn-Nardo Leslie
Published: January 2023

Background and Objective: On this literature review and textual analysis, we apply theories from health communication and psychology to better understand the cultural and structural factors that may discourage Black women’s participation in clinical trials, paying particular attention to the public and private organizations and agencies that fund this research. We use breast cancer clinical trials as an extended example to illustrate the wider issue of a lack of funding for formative and community-based research.
Design: We begin by reviewing the recruitment of Black women to breast cancer research and identifying themes in the literature where gaps exist. We then examine the social and organizational psychology literature on influencing the recruitment of women and racial/ethnic minorities, suggesting this field may be helpful for addressing the recognized gaps. Throughout the work, we identify points at which disparities may arise. We suggest that these theoretical approaches are helpful in reducing disparities and illuminating the structural factors that discourage participation from minoritized groups. Finally, we suggest future research opportunities for using these perspectives to increase participation of Black women in breast cancer and other health research.
Results: Researchers could benefit from employing psychology principles of contingency cues and identity safety when recruiting Black women into clinical trials, especially for breast cancer. Further, funding organizations such as the NIH must fund formative research which seeks out the voices of Black women and other marginalized populations to fully address and eliminate health disparities.
Conclusions: This work highlights the need for increased cross-sectional literature in the fields of communication and psychology. As scholars explore directions for future research, we encourage them to consider the expertise of Black women, themselves, in solving these problems.
Husisha: Training Peer-Counselors in Adolescent Problem-Solving Therapy in Eldoret, Kenya: Implementation, Adaptation, and Outcomes
Authors: Vander Missen Marissa , Barasa Julius , Jaguga Florence , Kwobah Edith , Turissini Matthew , Ott Mary A. , Puffer Eve , Kusgei Gilliane , Rono Wilter , Njiriri Faith , Giusto Ali
Published: January 2023

Background and Objective:In Kenya, youth needing mental health (MH) treatment vastly outnumber licensed mental health professionals. Task-sharing MH treatment to non-professionals has potential to reduce the treatment gap. For youth, non-professional peer counselors have the benefit of increasing engagement and reducing stigma. Problem-solving therapy (PST) is one treatment shown to alleviate MH symptoms even when delivered by non-professionals. Here we (1) evaluate the implementation of a PST training and (2) codify adaptations for PST implementation at a youth drop-in center at Family Health Options Kenya.
Experimental Design or Project Methods:A 2-week training for peer mentors was conducted. Curriculum included core counseling skills, overview of MH, and PST introduction and application.Peer Outcomes: Mean scores from pre- and post-written exams were compared using paired t-tests. Standard role plays were evaluated using the Working with children – Assessment of Competencies Tool (WeACT). Consensus WeACT scores from pre- and post-role plays were compared using paired t-tests.Adaptations: Feedback on cultural acceptability, language comprehensibility, and intervention flow was collected. Proposed intervention changes were coded within the Ecological Validity Model framework.
Results:Based on a written exam, mentors’ knowledge of MH conditions and core counseling skills improved after training (Pre: 10.88±4.36; Post: 15.38±2.88; p=.026). Competence in application of counseling skills evaluated with WeACT improved after training (Pre: 20.63±6.61; Post: 28±1.69). Examples of adaptations to PST include: changes to address stigma for MH treatment in Kenya and redistribution of content between sessions.
Conclusion and Potential Impact:A 2-week PST training improved peers' counseling skills and ability to deliver a manualized PST treatment. Training allowed contextual, conceptual, and methodological adaptations to PST for use in a Kenyan context. Implementing and improving lay-counselor trainings for MH interventions in Kenya has the potential to increase access to preliminary MH treatment.
Associations Between Psychiatric Symptoms and Cognitive Impairment in Post-Acute Sequelae of COVID
Authors: Todd Noah , Pater Jessica , Li Jason , Bohn Camden , Henriksen Brian , Carroll Jeanne , Chang Fen-Lei
Published: January 2023

Introduction: Post-acute sequelae of COVID (PASC) is an emerging postviral syndrome affecting patients who have recovered from acute COVID infection. Symptoms can affect many different organ systems, including the CNS. Neuropsychiatric complications are common in PASC, affecting roughly 1 in 3 patients who recover from COVID. The present study aims to investigate relationships between depressive, anxious, somatic, and general psychiatric symptom burden and cognitive impairment in PASC.
Methods: Patients were recruited from the Parkview Post-COVID Clinic (PPCC) from April 2021 to March 2022. Patients were deemed eligible if data were available for both Brief Symptomatic Inventory (BSI) and BrainCheck scores. Eligible patients were divided into low or normal cognitive function based on BrainCheck guidelines, and differences in psychiatric symptom severity were assessed via Mann-Whitney U testing. Strength and direction of associations were determined using Pearson’s correlation coefficient. A smaller subset of patients was analyzed for differences in PASC symptom severity based on Omicron or Delta infection.
Results: Generalized cognitive and mental flexibility complaints were associated with more severe psychiatric symptom burden in all domains measured by BSI (p = 0.003 and p = 0.01, respectively). Impaired attention was not related to any psychiatric symptom domain (p > 0.05). Impaired memory was related to more severe symptom burden in all psychiatric domains except depression (immediate recall p = 0.015; delayed recall p = 0.015).
Conclusion: General cognitive and mental flexibility complaints can be used to prompt clinical suspicion of concurrent psychiatric symptoms. Complaints regarding attention are not linked to any psychiatric symptom domain. Complaints regarding memory can also provide impetus for investigating concurrent psychiatric symptoms, excluding depression. These associations can be used to facilitate earlier diagnosis and treatment of cognitive impairment and psychiatric illness in the context of PASC.
Combinatorial Therapeutic Strategy of Stem Cell Retinal Organoids and Neurotropic Factor for Glaucoma
Authors: Ho Kathleen , Shahul Hameed Shahna , Sharma Tasneem
Published: January 2023

Purpose: Glaucoma is a group of optic neuropathies characterized by retinal ganglion cell (RGC) death and visual field loss. A degenerative mechanism associated with RGC death is disrupted delivery of neurotropic factors due to glaucomatous axonal damage. This transport is critical for protection of long-term neuronal function. Thus, we investigate a potential therapeutic target, human Neuritin 1 (NRN1), which has demonstrated neuroprotective effects in rodent axonal injury models. Further, the advent of induced pluripotent stem cell (iPSC) technology allows iPSC-RGCs to be generated in-vitro from commercial iPSCs and reprogrammed corneal fibroblasts. We hypothesize delivery of NRN1 and transplantation of iPSC-RGCs will sustain survival of RGCs and ultimately slow progression of glaucoma-induced neuronal death in our ocular translaminar autonomous system (TAS) perfusion model system. This will allow us to analyze a potential therapeutic approach for both early-stage glaucoma (NRN1 therapy) to protect dying RGCs, and late-stage (iPSC-RGCs), when most RGCs are lost.
Methods: Human donor eyes were obtained from eye banks according to Declaration of Helsinki. The iPSC-RGCs transfected with AAV2-GFP were seeded into human posterior cups with NRN1 and cultured in TAS model under pressurized conditions for 2 days. Survival of iPSC-RGCs, gliotic and fibrotic pathways were measured through expression by qRT-PCR and immunohistochemistry. Retinal function post-treatment was measured through electroretinogram analysis.
Results: We successfully maintained the human posterior eye cups in translaminar differentials for 2 days. In contrast to controls, we observed increased RGC survival and retinal function after combination therapy of NRN1 and iPSC-RGC. Additionally, we found differential gene expression of apoptosis, inflammation, and gliotic markers.
Conclusion: Our study identified that NRN1 in conjunction with iPSC-RGC transplantation treatment promotes RGC survival under glaucomatous conditions. This suggests that NRN1 and iPSC-RGCs could be utilized as a potential combination therapy to save retinal neurons and prevent neurodegeneration in glaucoma patients.
Tracking the Neurodegeneration and Behavioral Changes in Mice Model of Prodromal Phase Alpha-Synucleinopathy
Authors: Bari Yasin , Khan Hammad , Jayant Krishna
Published: January 2023

Background:
The aggregation of misfolded alpha-synuclein (αSyn) has been shown to cause neurodegeneration in a mouse model and lead to behavioral changes. While others have examined end-stage (after neurodegeneration has set in) behavioral changes in mice after αSyn preformed fibrils (PFF) seeding, the precise timeline of circuit changes and associated behavioral deficits during the prodromal phase of synucleinopathic spread is poorly understood. We set out to measure and characterize the extent to which behavior is affected by primary motor cortex (M1) PFF seeding in mice.
Experimental Design:
We quantified the neuronal cell body density at M1 layers 2/3 and 5 using immunohistochemistry cell assays to determine if there were neurodegenerative effects under the PFF seeding strategy. Furthermore, using Bonsai, a visual reactive programming language, we tracked the behavior of the PFF seeded mice for exploratory motives, decreases in which can be used to show cognitive decline. To further ascertain the effect of αSyn on vulnerable excitatory pathways in the cortex, we performed prodromal injections of αSyn PFF into the M1 of ChATCre mice along with projection tracing in the basal forebrain.
Results:
During the first 3 months following PFF seeding, the density of neuronal cell bodies in M1 layers 2/3 and 5 remains unchanged in PFF seeded mice compared to control mice. The exploratory index of PFF seeded mice was comparable to that of control mice. These results suggest insignificant neurodegeneration and behavioral change in the first 3 months following PFF seeding.
Conclusion:
Work done previously in this lab tracking the spread of PFF in M1 shows significant spread of αSyn for 3 months following prodromal injection. The results from this work show that there is little neurodegeneration or behavioral change in that timeframe despite the significant spread of αSyn. Future work hinges on further probing this phenomenon.
Evaluation of Gait Development Trajectory in Mice
Authors: Thayer Amanda , Villarreal Cameron , Chan Deva
Published: January 2023

Background/Objective: Movement and walking are huge facets in how we go about our lives and experience the world. Gait is compromised in numerous conditions, from injury and chronic pain to neurological conditions and arthritis. Movement biomechanics have therefore been studied extensively in humans for decades. Although animal models are often used to examine the progression of various health conditions, translational gait research in animal models are less complete, and there is limited information on gait changes as mice grow.
Methods: We recorded video segments of wild-type C57Bl/6J mice during skeletal growth (5 weeks to 16 weeks) walking (n = 4 Female, n = 2 Male) at 20 cm/s on the DigiGait system, a transparent treadmill equipped with a high-speed camera. Videos captured a ventral view of the mouse and were processed using DigiGait Analysis software, which uses local thresholding and position-finding algorithms to examine paw placement on the treadmill belt. Manual adjustments were made to eliminate artifacts and confirm heel-strike and toe-off times. After these corrections, the software returns stride parameters that were compared between sexes.
Results: Over 30 gait metrics, including stride length, frequency, stance/swing times, and stance width, were calculated for all four limbs per animal for several time points. Stride length increased from 4.5 ± .5 cm at week 5 to 5.0 ± .8 cm at week 16, and frequency decreased from 4.6 ± .6 to 4.2 ± .6 steps per second. No notable changes in stance width, symmetry, or stance/swing ratio were identified.
Conclusion and Potential Impact: Future directions for this research include conducting a principal component analysis to investigate the variability within the gait data. Identifying trends in gait parameters will help create a more complete picture of gait maturation. Establishing this baseline data allows for its comparison against treatment groups and the potentially meaningful investigation of translatable therapeutics and interventions.
Brain folding increases in sharpness and complexity over third trimester-equivalent development
Authors: Basinski Christopher , Garcia Kara
Published: January 2023

Background and Hypothesis:
Gyrification, or convolution, of the cerebral cortex is a promising transdiagnostic marker for early neurodevelopment. Previous studies have related differences in sulcogyral shape to schizophrenia, bipolar disorder, and autism spectrum disorder, but the physical mechanisms underlying these differences remain poorly understood. The focus of this study was to explore decomposed curvature metrics, the principal curvatures, as physically meaningful quantitative biomarkers to track brain development. We hypothesize that the average sharpness and complexity of sulci and gyri, reflected by principal curvatures, increase throughout third trimester-equivalent development.
Methods:
Cortical surfaces generated from magnetic resonance imaging (MRI) were obtained from the developing Human Connectome Project. Global sharpness was calculated from the principal curvature of maximum magnitude, with average sharpness defined separately for gyral (positive) and sulcal (negative) curvatures. Global complexity of folds (eg., curviness along the length of a fold) was calculated from variance in the principal curvature with minimum magnitude. Trajectory of each summary metric was fit over time using polynomial regression.
Results:
Forty-three subjects were removed due to incomplete curvature analysis or missing subject information, such that 541 preterm and term-born infants were evaluated with scan age ranging from 27 to 45 weeks postmenstrual age (PMA). Across this developmental range, sharpness and complexity increased until a plateau around term-equivalent. Average sharpness of gyri was best correlated with age of scan (R2 = 0.877).
Conclusion and Potential Impact:
During the pre- and postnatal development period, total cortical surface area continues to increase after birth, but the overall sharpness and complexity of folding plateaus at ~37 weeks post-menstrual age. Exploring these physically meaningful curvature metrics can provide improved parameters for comparison to mechanistic models of brain folding.
Evaluation of Gait Parameters using Wearable Sensors in Simulated Freezing of Gait Episodes
Authors: Szilagyi Halie , Yadav Amol P.
Published: January 2023

Background and Hypothesis: Freezing of gait (FoG), the failure to initiate or maintain locomotion, is a common symptom in Parkinsonism that severely impacts patients’ quality of life. Wearable technology such as inertial measurement units (IMU) are being widely investigated as a method of detecting these episodes with increasing sensitivity and specificity. The aim of this initial study is to collect objective gait data using IMU sensors via Perception Neuron software and to develop an analysis pipeline for quick in-clinic calculation of gait parameters.
Experimental Design: 16 IMU’s were attached to the feet, legs, arms, waist, back, and head in individuals without any gait abnormalities. Subjects were instructed to walk 10 meters normally, followed by 10 meters with interspersed, simulated FoG episodes. This data was processed and analyzed through Axis Studio and MATLAB to identify kinematic markers associated with gait. The correlation between the right and left foot gyroscopic data in the z-axis was determined and plotted against time, with any correlation less than 0.5 being considered a FoG episode. Further, parameters of gait including stride duration, stance duration, swing duration, and stride length were calculated from input of the raw data to establish a control comparison for future studies with Parkinson’s patients.
Results: Data analysis demonstrated increased stride duration, stance duration and swing duration with shorter stride length in the stimulated FoG episodes as compared to normal gait. The developed MATLAB code was further able to accurately distinguish the simulated FoG episodes from normal gait given the correlation parameter difference of <0.5.
Conclusion: Our findings showed that data obtained from Perception Neuron IMU can be processed to succinctly detect FoG episodes and calculate gait parameters. Ultimately, this data will be used as the baseline for a future study investigating the use of spinal cord stimulation as treatment for FoG in individuals with Parkinson’s Disease.
Ethanol Induces Blood-Brain Barrier Dysfunction in a Familial Alzheimer’s Human Stem Cell-Derived Model
Authors: Bell Kameron T. , Hughes Jason M. , Canfield Scott G.
Published: January 2023

Background:
Alcohol consumption has been directly linked to neurodegenerative diseases, including Alzheimer's disease (AD). Additionally, heavy consumption of alcohol has been shown to cause a faster cognitive decline in AD patients. A subset of studies demonstrated that alcohol can diminish BBB integrity and independently AD patients have suppressed barrier properties, but the direct effect of alcohol on barrier integrity in AD patients remains unclear. In this study, we utilize a human stem cell-derived AD BBB model with near in vivo properties to investigate the effects of alcohol on critical barrier properties.
Methods:
Brain microvascular endothelial cells (BMECs) were derived from healthy (IMR90) and AD (PSEN 1, PSEN 2, and APP) human induced pluripotent stem cells (iPSCs). Healthy and AD cell lines were treated with physiologically relevant concentrations of alcohol (5, 25, and 50 mM). Following exposure, several critical barrier properties were monitored for up to 5 days post-exposure, including trans-endothelial electrical resistance (TEER), sodium fluorescein permeability, and tight junction localization.
Results:
Moderate to severe alcohol exposure (25mM and 50mM) decreased barrier integrity in both healthy and AD-derived BMECs, as observed by an increase in sodium fluorescein permeability and a reduction in TEER. Furthermore, alcohol increased the number of discontinuous tight junctions directly contributing to the diminished barrier integrity. Interestingly, our preliminary results demonstrate that AD-derived BMECs are more susceptible to ethanol- induced barrier injury at lower concentrations of ethanol (5mM) compared to healthy-derived BMECs.
Conclusion and Potential Impact:
Our results indicate that alcohol can diminish critical barrier properties in healthy-derived BMECs similarly to other non-human established BBB models. For the first time, we observed an increase sensitivity to alcohol-induced BBB dysfunction in a familial AD-derived BBB model. These data suggest that mild alcohol consumption could significantly alter the BBB and contribute to the development or exacerbation of AD-induced barrier dysfunction.
Toxoplasma Gondii Induces Blood-Brain Barrier Dysfunction in a Human Stem Cell- Derived Model
Authors: Bell Kameron T. , Bitters Abigail , López-Yglesias Américo H. , Canfield Scott G.
Published: January 2023

Background:
Toxoplasma gondii is a single-celled obligate intracellular parasite commonly found in cat feces and animal issues and is known to cause cerebral infections in immune- compromised individuals and neonates. Once ingested, T. gondii reaches the host's blood supply and reaches the brain, where infection ensues. However, it remains unclear how T. gondii crosses the blood-brain barrier to reach the brain. In this study, we utilize a human stem cell-derived BBB model with near in vivo properties to investigate the effects of T. gondii on critical barrier properties that permit entry into the brain parenchyma.
Methods:
Human-induced pluripotent stem cells (iPSCs) were used to derive brain microvascular endothelial cells (BMECs). BMECs were exposed to tachyzoites, the infectious rapid replicating stage of T. gondii, at a multiplicity of infection (MOI) of 0.1 and 1.0 for 24 hours. Following exposure, several critical barrier properties were monitored, including trans- endothelial electrical resistance (TEER) and tight junction analysis.
Results:
BMECs treated with both MOI of 0.1 and 1.0 showed a significant reduction in all three tight junction proteins (Claudin-5, Occludin, and ZO-1), as indicated by an increase in discontinuous junctions and a decrease in area fraction index. Barrier integrity is directly associated with tight junctional continuity and localization. Furthermore, tachyzoites at an MOI of 0.1 and 1.0 demonstrated a reduction in barrier tightness, as observed by a decrease in TEER.
Conclusion and Potential Impact:
These results indicate that T. gondii may, in part, have access to the brain by inducing disruption in tight junction proteins, thus increasing barrier leakiness via a para-cellular route. These data may help guide future investigations on T. gondii's ability to evoke blood-brain barrier dysfunction and reach the vulnerable brain tissue.
Cellular Fractionation to Characterize the Interaction of Nucleolin with Alpha-COP
Authors: Iurillo Alyssa , Custer Sara , Androphy Elliot
Published: January 2023

Background and Hypothesis:The goal was to further characterize the interaction between Nucleolin and the alpha subunit of the COPI coatomer complex. Nucleolin contains a C-terminal dilysine motif, which mediates interactions between the WD40 domain of alpha-COP and COPI-interacting proteins. Previous work in the lab showed that this C-terminal dilysine is required for co-immunoprecipitation of alpha-COP with Nucleolin. Because alpha-COP is exclusively found in the cytoplasmic compartment, we hypothesized that the interaction between alpha-COP and Nucleolin is exclusively cytoplasmic but previous co-immunoprecipitations had only been performed from whole cell lysates. Alpha-COP has been shown to bind mRNA, but it was unclear whether this interaction was direct or whether alpha-COP was binding an RNA binding protein (RBP) which would act as a bridge between the mRNA and alpha-COP. Nucleolin acts as an RBP in both the nuclear and cytoplasmic compartments. We hypothesize that some of the mRNA bound to alpha-COP are present due to their association with Nucleolin.
Experimental Design or Project Methods:The first aim of the project was to optimize a reproducible cell fractionation protocol to reliably separate nuclear and cytoplasmic compartments. The second goal of the project was to identify mRNA that would co-immunoprecipitated with Nucleolin in HEK293T cells where we can easily express tagged versions of alpha-COP. Nucleolin-bound mRNA had previously been identified in Hela cells. We began by testing for the expression of these mRNAs in 293-TT cells using the published RT-PCR primers.
Results:After these tests identified Ftl (Ferritin light polypeptide) as a highly abundant transcript in 293-TTs, we performed RNA immunoprecipitation from cells expressing epitope tagged Nucleolin or alpha-COP. We confirmed that both Nucleolin and alpha-COP are in complex with Ftl mRNA.
Potential Impact:Future experiments will use short-hairpin RNA to knockdown Nucleolin and determine whether the levels of Ftl mRNA that co-immunoprecipitated with alpha-COP are reduced in the absence of Nucleolin.
Cellular Metabolism in B Cells in Type 1 Diabetes
Authors: Swar-Eldahab Mugtaba , Conway Holly , Felton Jamie L.
Published: January 2023

Background/Objective: Type 1 diabetes (T1D) is an immune-mediated disease that results in the destruction of pancreatic beta cells. While beta cell destruction is classically considered T cell mediated, autoreactive B cells play important roles in disease progression. B cell depletion prevents disease in non-obese diabetic (NOD) mice, and B cell depletion temporarily slows disease progression in individuals with new-onset T1D. However, mechanisms of autoreactive B cell function in T1D are not fully known. Cellular metabolism has been shown to drive autoimmune B cell development in other mouse models. We hypothesize that metabolic characteristics of B cells from NOD mice are distinct from metabolic characteristics of B cells from non-autoimmune C57BL/6J (B6) mice; therefore, making cellular metabolic pathways viable targets for therapeutic intervention.
Methods: Lymphocytes from spleen, pancreas, pancreatic lymph nodes, and mesenteric/lumbar lymph nodes were processed into single-cell suspensions. Glucose uptake was measured using fluorescent glucose analog 2-NBDG. Mitochondrial polarity was measured using fluorescent probes for mass and membrane potential. Cells were stained for surface markers and analyzed on an Attune Nxt flow cytometer.
Results: No statistically significant differences in glucose uptake or mitochondrial polarity for lymphocyte subsets in the spleen or PLNs of NOD and B6 mice were identified. In NOD mice, polarity was significantly higher in B cells in the pancreas compared to the spleen and PLNs. Polarity was also higher in B cells in PLNs compared to non-specific lymph nodes in NOD mice.
Conclusions/Impact: While no differences in glucose uptake or polarity in lymphocytes from NOD and B6 mice ex vivo were identified, future studies are needed to determine whether their activation drives metabolic alterations. Differences in polarity in the pancreas in NOD mice suggest that cellular metabolism is influenced by the islet microenvironment and has the potential to influence their function at the site of autoimmune attack.
Characterization of a conserved cysteine residue in the papillomavirus E2 protein
Authors: Stoll Kennedy , Gonzalez Jessica , Androphy Elliot J.
Published: January 2023

Background:Human papillomaviruses (HPVs) are DNA tumor viruses that infect cutaneous and mucosal epithelium. While most infections are self-limiting, a small subset that infects the mucosal epithelium progresses to cancer. All papillomaviruses encode the protein E2 which regulates viral transcription and replication; a highly conserved cysteine residue in the DNA contact helix of E2 plays an unknown role. Previous research suggests the residue is not necessary for replication or binding to DNA. We hypothesize that post-translational modification of this conserved cysteine residue leads to release of viral DNA during packaging of progeny virions.
Methods:Mutations of the murine papillomavirus conserved E2 C307 residue to serine and phenylalanine were used to investigate its role in E2 function. C33A, HPV negative cervical cancer cells, were transfected with an E2-responsive luciferase reporter and either wild type or mutant C307 E2 vectors; luciferase assays were performed 48 hours post-transfection to assess transcriptional activity. Whole cell lysates from overexpressed C307 mutants were separated by SDS-PAGE and immunoblotted to assess expression levels relative to wild type. To examine protein localization, C33A cells were transfected with equal amounts of wild type or mutant E2 and fixed 48 hours post-transfection for immunofluorescence.
Results:C307S and C307F mutants are both capable of weakly activating transcription. Overexpression of the mutants resulted in a dose dependent increase in transcriptional activity. Both mutants are expressed at levels comparable to wild type E2 and are correctly localized to the nucleus.
Conclusion/Impact:The deficient transcription function displayed by the C307 mutants cannot be explained by poor expression or mislocalization. Continued study of this conserved cysteine will help to further understanding of papillomavirus biology and may offer insight into novel avenues for treatment or prevention of HPV-associated cancers.
Microplastic Effects on Thrombin-Fibrinogen Clotting Dynamics Measured via Turbidity
Authors: Stelflug Nathan , Tran Daniela Q. , Hall Abigail , Chakravarthula Tanmaye Nallan , Alves Nathan J.
Published: January 2023

Background/Hypothesis:Widespread use of plastic has created a world where exposure to microplastics is inevitable leading to their presence in our circulatory system. This raises questions about microplastics impact on thrombosis. Aminated polystyrene (aPS) plastics have been shown to increase platelet aggregation and thrombus formation in animal models. Given this, we hypothesized that aPS administration will increase the rate of fibrin clot formation in a simple thrombin-fibrinogen clot model.
Project Methods:We evaluated how concentrations of 25-200 μg/mL of 100 nm aPS particles affect fibrin clot formation using turbidity assays. To determine the effect of surface charge, experiments were also performed with non-modified polystyrene (nPS) particles. Microplastics were pre-incubated either with physiological concentrations of fibrinogen or thrombin and the clot formation was measured using turbidity at 405 nm every 10 seconds over 45-minutes. Clotting parameters such as maximum turbidity (TurbMax), time to 90% maximum turbidity (TurbTime), and clot formation rate (Vmax) were determined and compared to controls without microplastics.
Results:When increasing concentrations of aPS were preincubated with thrombin or fibrinogen, there was less than a 2-fold change in Vmax, TurbMax, and TurbTime. When increasing concentrations of nPS were preincubated with thrombin, there was up to a 27-fold decrease in Vmax, 2.4-fold decrease in TurbMax, and 4.36-fold increase in TurbTime compared to the control. Whereas preincubation of nPS with fibrinogen resulted in 1.86-fold decrease in Vmax, 1.63-fold decrease in TurbMax, and 2.30-fold increase in TurbTime.
Potential Impact:In this simplified clotting model, it was surprising to find inhibitory effects on clot formation and that they were more pronounced with nPS than with aPS. However, these results align with increase prothrombin time observed in literature in presence of aPS. Therefore, future studies with more complex clotting models need to be performed before claims can be made on the impact of microplastics on thrombosis.
Validation of the Modified Radiographic Union Score for Tibia Fractures (mRUST) in Female Murine Femoral Fractures
Authors: Chen Larry , Lamb Nathan , Tucker Aamir , Nazzal Murad , Mostardo Sarah L. , Awosanya Olatundun , Blosser Rachel J. , Liu Ziyue , Kacena Melissa A.
Published: January 2023

Background: In the U.S, 1.1 million bone fractures require treatment each year. With bone fracture union predicting post-injury quality of life, investigation into therapies improving fracture outcomes is needed. Small animals like mice provide sound models of bone fracture healing. However, current in vivo methodology (μCT) assessing fractures in murine models is costly and may disturb fracture healing. The modified Radiographic Union Score for Tibia Fractures (mRUST) was developed to analyze human fractures efficiently using radiographs. This study hopes to demonstrate that mRUST provides comparable results to current standard measures of fracture healing in female murine models.
Methods: In 135, 12-week-old female C57BL/6 mice, femur fractures were induced by surgical procedure. Lateral radiographs were taken directly after surgery along with lateral and anterior posterior images at 7,10,14,17, 21, 24, 28, 35, and 42 days post-surgery (n=15/group). At each time point, 15 mice were sacrificed, and their bones collected for μCT, histomorphometry, and biomechanical analyses.   All radiographs will be randomized and assigned mRUST scores by orthopaedic surgeons blinded to groups. Callus and fracture line characteristics in four cortices of    radiographs dictate mRUST scores. The level of correlation between mRUST scores and current measures of fracture healing will be calculated.
Results: To date, all surgeries have been completed and all radiographs and specimens have been prepared for analyses. After analyses, statistical correlations between mRUST scores and μCT, histomorphometry, and biomechanics measures will be determined. We also plan to compare these findings to our previous findings with male mice to see if differences exist in fracture healing based on sex.
Conclusion: Recently, our group validated using mRUST scoring in male murine models. We predict the same for female models. This will bolster the use of radiographs to assess fracture condition in mice, providing a relatively inexpensive tool for fracture therapy assessment.
A Qualitative Study of a New Metric for Estimating the Risk of Early-onset Colorectal Cancer in Male Veterans
Authors: Cheng Michael , Matthias Marianne S. , Imperiale Thomas F.
Published: January 2023

Background: Identifying patients who benefit from screening prior to ages 45 or 50 through risk-prediction models may improve the uptake and effectiveness of colon cancer (CRC) screening. “Colon Age” is a way to estimate a person’s risk for CRC in Veterans younger than age 50. The purpose of this study is to determine what patients and primary care providers think about the “Colon Age” concept in terms of its comprehensibility, acceptability, and utility for decision-making about CRC screening.
Methods: Veteran patients < 50 years old and primary care providers (PCPs) were recruited from the Roudebush VAMC’s outpatient clinics. Semi-structured, qualitative interviews were conducted one-on-one and included Likert-scale and open-ended questions. The estimate of colon age is based on relating a person’s individual features to the 5-year age group in the U.S. population of men with the closest risk of CRC based on Surveillance, Epidemiology, and End Result cancer incidence data.
Results: 19 patients and 8 PCPs were recruited. The majority of patients (68%) had a colon age below that of their biological age, while 2 of 19 (11%) had a higher colon age. PCPs identified the tool’s potential to promote screening uptake, facilitate discussion between patients and PCPs, and adhere to current practices as facilitators. Identified barriers among PCPs and patients included questions about tool accuracy and validation, patient reluctance to receive any form of screening, and limited perceived utility for patients between the ages 45-49.
Conclusions and Potential Impact:  Using the concept of colon age to express individual patient risk for early-onset CRC was well-received among almost all veteran patients and physicians interviewed.Colon-age adapted screening tools   should be incorporated into theelectronic health record reminders to facilitate decision-making and screening uptake.Colon-age based personalized screening has the potential to improve cancer detection by providing individualized recommendations.
Circadian Rhythm Types in Individuals with Diabetic Macular Edema
Authors: Cory Matthew T. , Patel Janvi S. , Jusufbegovic Denis , Bhatwadekar Ashay
Published: January 2023

Background and Hypothesis:Diabetic Retinopathy (DR) is the leading cause of vision loss in patients with diabetes. Diabeticmacular edema (DME) can be associated with any stage of DR, causing thickening and swelling of  the  macula,  leading  to  vision  impairment.  Circadian  rhythm,  the  body’s  natural  clock  thatgoverns physiological processes and dictates our sleep-wake cycle, is known to be affected by DR and DME. This study hypothesizes that DME will be associated with a more extreme circadian rhythm type.
Methods:Individuals 18 years or older with a diagnosis of type I or type II diabetes complicated by DMEwere given a 19-item Morningness-Eveningness Questionnaire (MEQ) to determine their circadian rhythm and sleep  patterns. Information regarding demographics, metabolic parameters, diabetes, visual acuity and DME severity was collected.
Results:In  this ongoing study, we compiled data from 54 individuals out of a targeted enrollment of 160.  Overall,  the  MEQ  scores  characterized  more  individuals  as  morning  types,  followed  by  intermediate  and  evening  types.  The  preliminary  analysis shows  a  weak,  insignificant  positive correlation with retinal thickness in the right eye and a weak, insignificant negative in the left eye. In addition, a weak, insignificant negative correlation was also observed between MEQ and right eye visual acuity, with the opposite trend occurring in the left eye. DME variables such as mean central foveal thickness decreased insignificantly in right eyes from morning to evening groups but increased in the left eye.
Conclusion and Potential Impact:  Our study suggests DME individuals are mostly morning type instead of evening type. However, there is no strong relationship between MEQ scores and DME parameters. As we recruit more individuals in the future, we hope to establish a more significant relationship that could improve our understanding of how circadian rhythms and DME are interconnected and could lead to new treatments.
Longitudinal Study of COVID-19 Impact on Breast Cancer Screening in Northwest Indiana
Authors: Crowe Nicholas , Green Daniel , Han Amy
Published: January 2023

Background: During the COVID -19 pandemic, elective procedures such as mammography were suspended from late March to late April. Previous analyses done by Green and Han revealed that mammography screenings continued to be down in 2020 and 2021 from their baseline in 2019 prior to the pandemic. Continued efforts in monitoring how mammography screenings have been affected following the pandemic is crucial especially in understudied cities with predominantly black populations such as Gary, IN.
Methods: Data was provided from “Hospital A” on 22,693 mammography encounters and this data was analyzed to determine how mammography screenings have changed after the suspension period.  
Results: Following the lift on the suspension of elective procedures on April 27, 2020, there was a sharp increase of mammograms the following 8 weeks to near pre-pandemic levels. However, after this 8-week post suspension period of elective procedures on April 27, 2020, the weekly average began to decrease starting at the beginning of 2021. The weekly average went from 171 (SD 21.9) mammograms in 2019 to 108 (SD 40.0) mammograms in 2021. Furthermore, the weekly average has continued to decrease in 2022 with 101 (SD 46.8) mammograms.
Conclusion: “Hospital A” successfully returned mammography procedures to pre-pandemic levels initially during the 8 weeks following the suspension on elective procedures. However, strategies should be implemented to mitigate the continuing decrease in mammography procedures following the COVID-19 pandemic.
The Effect of Large Femoral Heads and Acetabular Cup Position on PROMs after Modern Posterior Approach THA
Authors: Darden Austin , Deckard Evan R. , Meneghini R. Michael
Published: January 2023

Introduction: Use of large femoral heads (≥40mm) in total hip arthroplasty (THA) decreases postoperative dislocation by increasing impingement-free range of motion, however, may leave patients more susceptible to groin pain.Also, limited data exist for the effect of large femoral heads and acetabular cup position on modern patient-reported outcome measures (PROMs). Therefore, the purpose of this study was to evaluate the effect of large femoral heads (≥40mm) and acetabular cup position on PROMs after primary THA.
Methods: 328 primary THAs performed by a single surgeon were retrospectively reviewed.Acetabular cup inclination and anteversion were measured using Martell Hip Analysis Suite software. Femoral head and acetabular cup sizes were recorded from the electronic medical record. Prospectively collected PROMs (and covariates) related to activity level, satisfaction, and overall hip health were evaluated.
Results: Age, covariates related to PROMs, and acetabular cup position did not differ between ≥40mm and <40mm femoral head groups (p≥0.177).The ≥40mm head group had significantly higher mean BMI and proportion of males (p≤0.022).UCLA Activity level and satisfaction scores did not differ preoperatively or postoperatively at 4-months or minimum 1-year follow-up between femoral head groups (p≥0.209 ). Preoperative HOOS JR scores did not differ by femoral head groups (p=0.538).At 4-months, mean HOOS JR score was significantly higher in the ≥40mm head group compared to the <40mm head group (p=0.027); however, both groups achieved similar mean HOOS JR scores by minimum 1-year follow-up (p=0.956). HOOS JR score >90 and being ‘very satisfied or satisfied’ correlated with wide ranges and several combinations of acetabular cup inclination and anteversion.
Conclusion: Patients achieved comparable PROMs regardless of femoral head size suggesting large femoral heads may not leave patients susceptible to groin pain in addition to reducing the risk of postoperative dislocation. Excellent patient outcomes correlated with wide ranges of acetabular cup position.
Complications Related to Prolonged Use of Topical Steroids Among Children with Uveitis: A retrospective study
Authors: Edwards Cameron , Taylor Nicole M. , Tarvin Stacey E. , Rodriguez Martha
Published: January 2023

Background: Uveitis can be described as inflammation of the eye. It is classified depending on the location of the inflammation in anterior, intermediate, posterior and panuveitis. The current guidelines for first-line treatment include glucocorticoids, topical with eyedrops, intraocular corticosteroid injections and systemic (oral). Potential complications of patients on prolonged topical steroids include, glaucoma, and cataracts, which can subsequently cause permanent visual loss. The aim of this study is to describe the complications related to the use of prolonged topical steroids among pediatric uveitis patients.
Methods: This retrospective observational study reviewed the medical records of children (age ≤ 18 years) with uveitis at Riley Hospital for Children between 1997-2022 data collected included age at onset of uveitis, sex, anatomic location of ocular inflammation, comorbid disease (including systemic inflammatory disease), ocular complications, relevant laboratory data, and treatment. Study data was collected and managed using REDCap. Statistical analysis was performed using SPSS software.
Results: Out of 122 subjects, anterior uveitis constituted 45.1% of patients, intermediate 15.6%and posterior uveitis 0.8%.  The median age of diagnosis was 10 years of age and 53.3% were female. Among these 54.1% of patients were placed on steroids, of which 49.2% were on topical steroids drops and 5.7% on oral steroids. Within the group of patients on chronic steroids 18% developed cataracts and 8.2% subsequently required corrective surgery, 10.7% developedglaucoma and 13.9% required intraocular pressure reducing drops. `
Conclusion and Potential Impact:  Uveitis treatment can be challenging, and some patients require longer periods of topical steroid use, which can be associated with complications including glaucoma and cataracts which can lead to blindness. In this cohort 18% developed cataracts and 10.7 % developed glaucoma. These effects can be mitigated with attentive monitoring. Balancing the benefits and risks of steroids and judicious use are vital to optimizing ocular health.
A Mineralized Allograft in Lumbar Interbody Fusion – A Retrospective Chart Review Study
Authors: Fiechter Jay , Smith Micah
Published: January 2023

Background: Lumbar interbody fusion is a routine surgical intervention for treatment of degenerative disc disease to treat lumbar stenosis. Minimally invasive surgical approaches do not afford the opportunity to harvest large amounts of local bone autograft, therefore iliac crest autograft and or additional allograft material is strongly relied upon to achieve an arthrodesis. This study uses a blend of a blend of Ventris Medical’s osteoinductive allograft; Allocell® and SurGenTec’s nanoputty OsteoFlo® as an effective autograft extension material.
Methods: A retrospective chart review was performed on patients that underwent lumbar interbody fusion between October 2021 and January 2022. Demographic information was recorded, three- and six-month follow-up radiographs were assessed, and arthrodesis was graded using the Bridwell Anterior Grading System.
Results: A total of 18 patients were included in the study, representing 27 lumbar level fusions. A Bridwell grade of I (successful fusion) was achieved at three months in 11.1% of cases and at six months in 85.2% of cases. Four fusions remained a Bridwell grade of II at six months and were subsequently evaluated for complete arthrodesis. No patients received a Bridwell grade of IV (lucency with collapse of graft) at three- or six-month follow-up.
Conclusions: The combination of OsteoFlo® and Allocell® contributed to successful arthrodesis status post lumbar fusion.
EPHA2 is a Potential Target for the Treatment of NF2-/-Vestibular Schwannoma
Authors: Foster Kéyana , Mitchell Dana K. , Flint Alyssa , Rodriguez Brooke , Mang Henry , Davis Chris , Angus Steven P. , Clapp D. Wade , Yates Charles
Published: January 2023

Neurofibromatosis type 2 (NF2) is an autosomal dominant cancer predisposition syndrome characterized by the development of bilateral vestibular (VS) and spinal schwannomas secondary to loss of heterozygosity of NF2 in Schwann cells or their precursors. While these tumors are largely benign, they can cause considerable morbidity due to compromised auditory, vestibular, facial, and vertebral nerve function. This may result in deafness, vertigo, facial muscle weakness, chronic neuropathic pain, and even death.There are currently no pharmacotherapies for VS, and surgical resection remains the standard of care, which is associated with significant morbidity. Thus, there is an urgent need to develop pharmaceutical approaches to halt or reverse the progression of tumor growth in NF2 patients who develop VS. Our lab previously identified the receptor tyrosine kinase inhibitors brigatinib and dasatinib as potentially efficacious agents for the treatment of VS and demonstrated that both agents targeted the Ephrin A2 receptor (EPHA2). EPHA2is a transmembrane receptor tyrosine kinase thatis involved in cell contact-mediated motility, adhesion,and migration. Additionally, EPHA2 modulates axon guidance, and synaptogenesis in developing brain. Here we demonstrate that EPHA2expression is increased in NF2-/-Schwanncells and NF2-/-cancers. We identify ponatinib, a receptor tyrosine kinase inhibitor targeting ABL1 that is FDA-approved for CML, as an additional agent that targets EPHA2. We demonstrate that ponatinib treatment impairs the viability of both human and murine NF2-/-Schwanncells in vitro and decreases EPHA2 protein expression. Accordingly, pharmacologic,and siRNA-mediated inhibition of EPHA2 also impaired the growth of human NF2-/-Schwann cells in vitro. Lastly, we demonstrate that both ponatinib and EPHA2 inhibition induce morphological changes in NF2-/-Schwann cells. Our findings suggest that ponatinib or the direct targeting of EPHA2 may be efficacious for the treatment of NF2-associated vestibular schwannoma. Future in vivo efficacystudies are warranted.