bluuebunny/crossref_metadata_2025_split huggingface dataset:
Evaluating the effects of RAS and CK2 Inhibitors in ESR1 Mutated Breast Cancer
Authors: Suescum Camilo , Nakshatri Harikrishna 
Published: December 2021

Background and Hypothesis:
Estrogen Receptor alpha (ESR1) is rarely mutated in primary breast cancers but is frequently mutated in metastasis that can appear after many years of anti-estrogen therapy. Mutations to ESR1 can result in estrogen-independent activity of ESR1 causing anti-estrogen to become ineffective. Previous work on this project has led to the hypothesis that RAS pathway activation in metastatic cancer cells as a result of ESR1 mutation leads to elevated CK2 activity which ultimately results in metastatic progression. Therefore, we hypothesize that the use of RAS signaling inhibitors or CK2 inhibitors have efficacy in blocking or reducing the metastatic progression of metastatic breast cancers with hyperactive RAS pathways.
Experimental Design or Project Methods:
The estrogen receptor positive, anti-estrogen sensitive breast cancer cell line MCF-7 and the same cell line genomically modified to replace wild type ESR1 to breast cancer metastasis enriched Y537S or D538G ESR1 mutation were used in this study. Cells were treated with various concentrations of the RAS pathway inhibitor Salirasib or CK2 inhibitor Silmitasertib and cell proliferation rates were measured using bromodeoxyuridine incorporation ELISA.
Results:
Thus far, the use of RAS signaling inhibitors or CK2 inhibitors have not shown efficacy in decreasing the proliferation rates of modified ESR1 MCF-7 cells. While there is a general trend of growth inhibition by these inhibitors at a higher concentration, there is no significant difference between the ESR1 mutant expressing cells and their respective controls.
Conclusion and Potential Impact:
This study will establish the feasibility of using RAS signaling inhibitors or CK2 inhibitors in the treatment of metastatic estrogen receptor-positive breast cancer. Future studies testing the effects of these drugs either alone or in combination with the clinically used anti-estrogen Fulvestrant for not only primary tumor growth but also metastasis in clinically relevant in vivo models may ultimately lead to clinical translation. Finally, demonstrating efficacy in these types of drugs may fuel the further refinement of drugs targeting these pathways to treat metastatic breast cancer.
Metabolic Syndrome and SIRT1 Mutation Impair Ca2+ Channels in Coronary Smooth Muscle Cells of Ossabaw Miniature Swine
Authors: Thamba Aish , Reed John , Strobel John S. , Byrd James , Alloosh Mouhamad , Coutts Alex , Sturek Michael 
Published: December 2021

Background:
Changes in Ca2+ regulation have been implicated in various pathologies such as coronary artery disease and metabolic syndrome (MetS), thereby potentiating these diseases. Our lab has shown that MetS decreases voltage-gated Ca2+ channel (VGCC) activity and sarcoplasmic reticulum (SR) Ca2+ release in coronary smooth muscle cells and increases coronary artery disease in Ossabaw miniature swine. Furthermore, decreased SIRT1 enzyme function can impair Ca2+ signaling and increase coronary disease and MetS. We hypothesized that impaired SIRT1 and MetS would decrease VGCC function and SR calcium store.
Methods:
CRISPR/Cas9 methods delivered a leucine to proline point mutation in SIRT1 (SIRT1L100P) into the Ossabaw swine genome to compare to wild type (WT), mimicking the naturally occurring mutation in humans which decreases SIRT1 activity. Four treatment groups of juvenile swine were based on genotype and diet: WT Lean, SIRT1 Lean, WT MetS, and SIRT1 MetS. Lean swine were fed normal chow and MetS were fed a hypercaloric, atherogenic diet for 7 months. The left anterior descending coronary artery was harvested and enzymatically digested to obtain cells. Fluorescence microscopy measured the Ca2+ indicator fura-2 in single cells. Depolarization of cells with perfusion of 80 mM K+ was used to elicit Ca2+ influx through VGCC. Caffeine (5 mM) exposure activated the Ca2+ release channel (ryanodine receptor) on the SR.
Results:
MetS was confirmed by increased body weight, impaired glucose tolerance, hyperinsulinemia, and hypercholesterolemia. Coronary atherosclerosis was shown by angiography, intravascular ultrasound, and gross imaging. A two-way analysis of variance revealed statistically significant overall effects of genotype (p=0.02), diet (p<0.0001), and an interaction (p<0.0001) between these variables to decrease VGCC function. In contrast, no effect was observed on SR Ca2+ release.
Conclusion and Potential Impact:
SIRT1 inhibition and MetS decreased VGCC function independently, but not additively or synergistically. (Support: NIH T35HL110854, DK120240, DK09751.)
Using Lyve1-Cre+ Mice to Visualize Lymphatics for Laser Capture Microdissection
Authors: Patel Aadarsh , Mohan Ganesh , Khan Imran , Sinha Mithun , Hassanein Aladdin H. 
Published: December 2021

Background:
Lymphedema is characterized by limb swelling secondary to lymphatic dysfunction. Lymphedema most frequently develops following breast cancer treatment due to iatrogenic damage of the lymphatics from surgery and radiation. Lymphedema affects 20-40% of breast cancer survivors. There is no cure for this disease. For determination of successful delivery of gene-based therapies, target cells are often isolated and analyzed via real-time PCR. One method to isolate a region of cells within a tissue section is laser capture microdissection (LCM). This process involves outlining the desired regions, which are cut on membrane slides and captured using a laser. Using LCM requires visualization and identification of the target tissue. In the case of lymphedema therapies, the target tissue is lymphatic vasculature.

Rationale of Project:
While lymphatics can be visualized using immunohistochemistry antibodies specific to lymphatic markers, the process is time consuming and can interfere with RNA levels in the tissue. Another option to visualize lymphatics is to use Lyve1-Cre+ mice. These mice express enhanced green fluorescent protein on lymphatic cells. The purpose of this project is to assess the utility of Lyve1-Cre+ and develop the methodology to capture the lymphatics in the murine tail to enable utilization of this methodology in murine tail model of lymphedema.

Methodology Development:
Samples of Lyve1-Cre+ mouse tails were harvested. Sections (10µm) were captured on LCM slides and dehydrated. The slides were visualized on the LCM microscope system, lymphatics vessels fluoresced green. They were captured via laser dissection. The captured samples were analyzed for the lymphatic specific genes (Prox1 and Lyve1) via quantitative real-time PCR to determine the purity of capture.
Child Abuse Screening in the Emergency Department Before and After Implementation of a Formal Screening Tool
Authors: Robertson Erika , Kanis Jessica 
Published: December 2021

Background:
Child abuse and neglect is a leading cause of mortality and morbidity in children. Victims of abuse often present to emergency departments due to these injuries. Early identification of suspected child abuse by emergency department staff allows the opportunity for intervention prior to a devastating injury. Signs of child abuse can be subtle and may not always be identified. Recent efforts have worked to improve suspected child abuse identification by implementation of validated screening protocols. Specifically, the IU Health Riley Emergency Department has recently implemented a new child abuse screening tool (effective on 15Apr2021). The Covid-19 pandemic has further exacerbated this issue, creating social isolation and additional socioeconomic stressors that contribute to child abuse.
Method:
A retrospective chart review was conducted to analyze patients identified for child abuse evaluation in the Riley emergency department. Patients prior to the implementation of the screening tool were identified by those undergoing a social work evaluation for child abuse. For patients presenting after implementation of the screening tool, all patients with a positive screen were reviewed. Charts were then reviewed for baseline demographics, diagnostic testing performed, and injuries identified. All data was recorded in an electronic Redcap database.
Results:
Implementation of a formal screening tool increased the number of patients identified for a child abuse evaluation (1.34% vs 3.17%) and those evaluated who had an injury also increased (1.15% vs 1.81%). More patients received a head CT scan after implementation of the screening tool (0.89% vs 1.07%), while the percentage of patients with an abnormal head CT decreased (62% vs 45%).
Conclusions:
Implementation of a formal screening tool for child abuse in the emergency department successfully improved identification of patients that should be further evaluated for child abuse. These results may be used to support implementation of the tool in additional emergency departments.
Impact of Gabapentin and Pregabalin on Neurological Outcome After Ischemic Stroke
Authors: Weber Michael , Morton Caleb , Chang Fen-Lei 
Published: December 2021

Background
The purpose of this study is to determine whether patients taking either gabapentin or pregabalin at the time of their stroke injury tend to have better outcomes than patients with similar injuries who were not taking one of the two medications. Prior studies have shown a potential neuro-protective effects of these two medications.
Methods
A retrospective chart review of 115 ischemic stroke patients from 2016-2021 were assessed for patient outcomes using two tools, the NIH Stroke Scale (NIHSS) and the modified Rankin Scale (mRS), in addition to their hospital length of stay. The outcomes of patients taking either gabapentin or pregabalin with stroke diagnoses are compared to patients with stroke diagnoses who were not taking either medication. Kruskal-Wallace and X2 were used for statistical analysis.
Results
There was significantly larger proportion of gabapentin patients that improved compared to patients in the control group when using the mRS tool for patient outcomes (X2; p=0.015). The gabapentin group showed a significantly larger improvement in the NIHSS scores from admission to discharge (Kruskal-Wallace; p=0.0005). Patients on gabapentin had a longer hospital stay than those not taking the medication by 1.7 days (t-test; p=0.041).
Conclusion
Our data support the potential neuro-protective effect of gabapentin/pregabalin with improved outcomes after an ischemic stroke using two parallel outcome measures of NIHSS and mRS scores. Of interest, patient hospital stays were longer on gabapentin/pregabalin, which may contribute to the improved outcomes. We need larger subject groups to confirm and further study our findings. This often can be facilitated by studies involving larger medical practices, insurance, or payer databases. In addition, impact of associated cost and care quality issues such as nosocomial infection and fall risk can be considered in the context of healthcare integration and value-based care emphasizing quality and cost management.
MicroRNASeq Target Analysis and Validation by Real-Time PCR in Abdominal Aortic Aneurysm
Authors: Sasso Willa , Moldovan Leni , Murphy Michael 
Published: December 2021

Background/ Objective: Abdominal aortic aneurysm (AAA) is an epigenetic event characterized by chronic inflammation and degeneration of the aortic wall leading to catastrophic rupture. Cigarette smoke exposure is the greatest environmental risk factor associated with AAA development. MicroRNAs (miRNA) regulate gene expression and may play a role in smoking-induced aortic inflammation. Epigenetic changes could include dysregulation of miRNA, causing post-transcriptional abnormalities pathogenic to AAA.

Methods: miRNA was extracted from plasma of 24 AAA patients and 7 risk factor matched (RFM) patients and analyzed by RNA sequencing. We compared previous (PS) and current smokers (CS) within and between both patient cohorts. Differential expression of miRNAs was analyzed by ANOVA (p≤ 0.05). Potential targets of significant differentially expressed miRNAs were predicted using cross-analysis of TargetScan and miRanda databases.

Results: Analysis revealed 7 significantly different miRNAs between AAA CS and AAA PS and 6 significantly different miRNAs between RFM CS and RFM PS. Of greatest significance, hsa-miR-223-3p was significantly downregulated as an effect of smoking cessation in AAA PS compared to AAA CS (p=0.000263), while also showing clinically relevant expression levels. Target genes of hsa-miR-223-3p include pro-inflammatory factors IL-6, TNFα, TGFβ, and MCP-1. Speculatively, as tissue levels of miR-223 tend to inversely correlate with plasma levels, we could hypothesize that the observed plasma upregulation of hsa-miR-223-3p in AAA CS contributes to the pro-inflammatory microenvironment of aortic tissue.

Conclusion: Cigarette smoke contributes to epigenetic changes impacting factors of immune regulation or inflammation, eventually leading to disease states such as AAA. Inflammatory-related hsa-miR-223-3p is upregulated in AAA CS, suggesting its potential role in the disease course.

Implications: Upregulation of hsa-miR-223-3p in AAA CS offers a link between disease state and the number one environmental factor attributed to AAA. This signature miRNA could serve as a biomarker for AAA or as a potential therapy target.
Three-dimensional morphometric analysis of human kidney nephron structures in health and disease
Authors: Bhaskara Suneil , Ferkowicz Michael , Barwinska Daria , Ashkar (El-Achkar) Tarek M. 
Published: December 2021

Background and Hypothesis: Chronic kidney disease (CKD) is very common and affects as many as 37 million people in the United States. Diabetes and hypertension are the most common causes of CKD. The pathogenesis of CKD is not fully elucidated. Morphological changes such as glomerulosclerosis and tubular atrophy are commonly observed with advanced disease. However, it is unclear if such changes occur at earlier stages of disease, a time when therapeutic interventions are likely to have the most benefit. Measurements of glomerular, vascular and tubular dimensions have been typically derived from thin section, but a pipeline for acquiring these morphometric measurements in 3-dimentions have not been performed. Using enhanced tissue clearing, confocal 3D imaging and volumetric segmentation and analysis, we aimed at developing an approach to measure the dimensions of various structures within the human kidney. Our overarching hypothesis is that such approach could allow the detection of morphometric changes in early disease.
Experimental Design or Project Methods: Kidney tissue were used either from donor nephrectomies or stored frozen biopsies from Biobank Biopsy Cohort of Indiana (BBCI). Enhanced tissue clearing and staining for nuclei, endothelium and proximal tubules were performed. Optical sectioning and 3D Imaging was done using confocal microscopy. Volume rendering, segmentation and analysis were done using the Imaris Software (Bitplane). Various built in and customized segmentation approaches were used.
Results: Volumes spanning 200 µm in thickness encompassing entire glomeruli and tubules were obtained. 3D Rendering of these volumes allowed visualization and enabled 3D segmentation. Dimensions of entire human glomeruli (maximum and minimum diameters, glomerular volumes and cellular densities), tubules and vessels were defined in control and samples with diabetic kidney disease.
Conclusion and Potential Impact: Our studies established an approach for accurate measurements of the dimensions of nephronal structures preserved in their 3D environment within tissue. We also established the feasibility of this approach in comparing changes with disease vs. control. Such methodology will open up a new line of investigations to better understand the pathogenesis of CKD, particularly at its early stages.
The Association of ADRA2A Genotype with Dexmedetomidine Dosage in Pediatric Patients
Authors: Biju Ashwin , Gallaway Katherine A. , Skaar Todd C. , Tillman Emma M. 
Published: December 2021

Background: Critically ill children in the pediatric intensive care unit (PICU) often require sedation with dexmedetomidine, an α2-adrenergic receptor (ADRA2A) agonist. In adults, evidence suggests an ADRA2A polymorphism (rs1800544) modulates the dexmedetomidine response. We hypothesize that pediatric patients with at least one wild-type allele (GG and GC) will require larger dexmedetomidine doses and a longer time to effective dose than patients with a variant genotype (CC).
Methods: This study was approved by the institutional review board (IRB). Patients in the PICU, 1 week through 25 years of age, and sedated with dexmedetomidine for ≥ 2 days were approached for enrollment. Genomic DNA was extracted from blood or saliva before genotyping for rs1800544 with a custom designed TaqMan® Assay on the QuantStudio 12K Flex platform. Patient genotype was determined using TaqMan® Genotyper software. Patient data were collected from the electronic medical record.
Results: Sixty-five patients were enrolled and genotyped. Twenty-six patients were homozygous variant (CC), 27 heterozygous (GC), and 12 wildtype (GG) for rs1800544. The maximum dose (mcg/kg/hr) was 0.75 ± 0.31, and 0.78 ± 0.32, in patients with and without a G allele, respectively (p=0.65). The mean doses were 0.59 ± 0.22 and 0.58 ± 0.24 in patients with and without a G allele, respectively (p=0.85). The time to effective dose (hrs) was 5.8 ± 12.0 for patients with a G allele and 4.0 ± 8.2 for patients with no G allele (p=0.74).
Conclusion:  Our hypothesis that the CC genotype would require lower doses of dexmedetomidine was not confirmed. Patients without a G allele required lower dexmedetomidine dosage and a longer time to achieve an effective dose, but these findings were not significant. This could be due to the small sample size or clinical factors that affect pediatric sedation. An adequately powered study could determine the association of ADRA2A genotype with dexmedetomidine.
Effect of CKD-MBD therapies on the gastrointestinal expression of phosphate transporters
Authors: Carlton Olivia , Chen Neal , Moe Sharon , Biruete Annabel 
Published: December 2021

Background: Chronic kidney disease-mineral and bone disorder (CKD-MBD) is prevalent and encompasses biochemical abnormalities, bone alterations, and vascular calcifications. CKD-MBD treatments include phosphate binders and calcimimetics that effectively lower phosphorus and PTH, respectively, but the impact of these treatments on phosphate transporters in the gastrointestinal tract is still unknown. NaPi2B is considered the primary intestinal phosphate transporter. However, NaPi2B inhibition shows limited effectiveness, thus the importance of PIT1 and PIT2 requires further investigation.
Methods: We tested the effects phosphate binders (ferric citrate (FC) and calcium gluconate (Ca)) and the calcimimetic KP2326 (KP) in (Cy/+ male rat; CKD) and untreated normal littermates (NL). Treatments lasted 10 weeks until euthanasia at 28 weeks (moderate-to-advanced CKD), where we collected mucosa samples from duodenum, jejunum, and ileum. Blood was collected for biochemistry measurements. Total RNA was isolated, and qPCR performed to assess phosphate transporters expression (NaPi2B, PIT1, PIT2) normalized to b-actin. Results were analyzed via 2-way ANOVA.
Results: As expected, CKD had abnormal plasma concentrations of phosphorus, PTH, and FGF23. FC and KP effectively lowered phosphorus. KP and Ca lowered PTH, but Ca increased FGF23. NaPi2B was expressed in the duodenum and jejunum but not in the ileum, and its expression was upregulated with FC compared to NL. PIT1 was expressed in all segments, but the expression was the highest in the ileum, while PIT2 was constitutively expressed in all segments. Ca led to higher PIT1 and PIT2 expression in the duodenum and jejunum compared to NL, CKD, or KP. KP led to higher expression of PIT1 in the ileum compared to CKD, FC, and Ca.
Conclusions: CKD-MBD therapies differentially impacted biochemistries and phosphate transporters expression. The effect of Ca on gastrointestinal expression of PIT1 and PIT2 may explain the higher plasma phosphorus and should be further explored.
Impact of Neonatal Outcome on Language Used to Describe Periviable Birth Experience
Authors: Casbon Courtney , Patel Meera , Tucker Edmonds Brownsyne 
Published: December 2021

Motivation and Purpose: The experience of giving birth to a periviable neonate between 22-25 weeks gestation varies greatly among women depending on numerous factors. Studying periviable narrative experiences will allow us to isolate those factors that create a more positive experience, even in the instance of a negative neonatal outcome (infant death). Results may be used to advise future caregiving approaches so they may be tailored to the needs of women in these situations. This study aimed to qualitatively assess the extent of the relationship between neonatal outcome and the language used by women and their important others (IOs) to describe periviable birth experience three months postpartum.
Method: A semi-structured interview guide was used to investigate participants’ perceptions of their delivery experience and subsequent NICU experience, if applicable, at three months postpartum. Interviews were transcribed verbatim and coded for themes by three investigators. Conventional content analysis was conducted using NVivo 12.
Results: Twenty-three women and twelve IOs were recruited at two hospitals between 2016 and 2018. Women and IOs who experienced infant death negatively described their delivery experience if they also negatively described healthcare team (HCT) interactions, including providers’ communication, clinical judgement, and personality traits. Specifically, participants with overall negative birth experiences described not understanding various clinical decisions and felt uninformed on topics pertinent to their or their infant’s care. Alternatively, women and IOs who experienced infant death who positively described their delivery experience also positively described NICU care (including HCT interactions) and infant death experience (comfort care). These results highlight the impact of HCT interactions and communication on a patient’s perception of delivery despite a negative neonatal outcome.
Conclusion: These findings could suggest that improvements in communication and information dissemination from the HCT may have the ability to improve a patient’s periviable birth experience despite a negative neonatal outcome.