bluuebunny/crossref_metadata_2025 huggingface dataset:
Telomerase Reverse Transcriptase and p53 Regulate Mammalian Peripheral Nervous System and CNS Axon Regeneration Downstream of c-Myc
Authors: Ma Jin-Jin , Ju Xin , Xu Ren-Jie , Wang Wei-Hua , Luo Zong-Ping , Liu Chang-Mei , Yang Lei , Li Bin , Chen Jian-Quan , Meng Bin , Yang Hui-Lin , Zhou Feng-Quan , Saijilafu
Published: October 2019

Although several genes have been identified to promote axon regeneration in the CNS, our understanding of the molecular mechanisms by which mammalian axon regeneration is regulated is still limited and fragmented. Here by using female mouse sensory axon and optic nerve regeneration as model systems, we reveal an unexpected role of telomerase reverse transcriptase (TERT) in regulation of axon regeneration. We also provide evidence that TERT and p53 act downstream of c-Myc to control sensory axon regeneration. More importantly, overexpression of p53 in sensory neurons and retinal ganglion cells is sufficient to promote sensory axon and optic never regeneration, respectively. The study reveals a novel c-Myc-TERT-p53 signaling pathway, expanding horizons for novel approaches promoting CNS axon regeneration.

SIGNIFICANCE STATEMENTDespite significant progress during the past decade, our understanding of the molecular mechanisms by which mammalian CNS axon regeneration is regulated is still fragmented. By using sensory axon and optic nerve regeneration as model systems, the study revealed an unexpected role of telomerase reverse transcriptase (TERT) in regulation of axon regeneration. The results also delineated a c-Myc-TERT-p53 pathway in controlling axon growth. Last, our results demonstrated that p53 alone was sufficient to promote sensory axon and optic nerve regenerationin vivo. Collectively, the study not only revealed a new mechanisms underlying mammalian axon regeneration, but also expanded the pool of potential targets that can be manipulated to enhance CNS axon regeneration.
Microglial P2Y12 Receptor Regulates Seizure-Induced Neurogenesis and Immature Neuronal Projections
Authors: Mo Mingshu , Eyo Ukpong B. , Xie Manling , Peng Jiyun , Bosco Dale B. , Umpierre Anthony D. , Zhu Xiaoqin , Tian Dai-Shi , Xu Pingyi , Wu Long-Jun
Published: October 2019

Seizures are common in humans with various etiologies ranging from congenital aberrations to acute injuries that alter the normal balance of brain excitation and inhibition. A notable consequence of seizures is the induction of aberrant neurogenesis and increased immature neuronal projections. However, regulatory mechanisms governing these features during epilepsy development are not fully understood. Recent studies show that microglia, the brain's resident immune cell, contribute to normal neurogenesis and regulate seizure phenotypes. However, the role of microglia in aberrant neurogenic seizure contexts has not been adequately investigated. To address this question, we coupled the intracerebroventricular kainic acid model with current pharmacogenetic approaches to eliminate microglia in male mice. We show that microglia promote seizure-induced neurogenesis and subsequent seizure-induced immature neuronal projections above and below the pyramidal neurons between the DG and the CA3 regions. Furthermore, we identify microglial P2Y12 receptors (P2Y12R) as a participant in this neurogenic process. Together, our results implicate microglial P2Y12R signaling in epileptogenesis and provide further evidence for targeting microglia in general and microglial P2Y12R in specific to ameliorate proepileptogenic processes.

SIGNIFICANCE STATEMENTEpileptogenesis is a process by which the brain develops epilepsy. Several processes have been identified that confer the brain with such epileptic characteristics, including aberrant neurogenesis and increased immature neuronal projections. Understanding the mechanisms that promote such changes is critical in developing therapies to adequately restrain epileptogenesis. We investigated the role of purinergic P2Y12 receptors selectively expressed by microglia, the resident brain immune cells. We report, for the first time, that microglia in general and microglial P2Y12 receptors in specific promote both aberrant neurogenesis and increased immature neuronal projections. These results indicate that microglia enhance epileptogenesis by promoting these processes and suggest that targeting this immune axis could be a novel therapeutic strategy in the clinic.
The Drosophila CPEB Protein Orb Specifies Oocyte Fate by a 3′UTR-Dependent Autoregulatory Loop
Authors: Barr Justinn , Gilmutdinov Rudolf , Wang Linus , Shidlovskii Yulii , Schedl Paul
Published: October 2019

orb encodes one of the two fly CPEB proteins. These widely conserved proteins bind to the 3′UTRs of target messenger RNAs (mRNAs) and activate or repress their translation. We show here that a positive autoregulatory loop driven by the orb gene propels the specification of oocyte identity in Drosophila egg chambers. Oocyte fate specification is mediated by a 3′UTR-dependent mechanism that concentrates orb mRNAs and proteins in one of the two pro-oocytes in the 16-cell germline cyst. When the orb 3′UTR is deleted, orb mRNA and protein fail to localize and all 16 cells become nurse cells. In wild type, the oocyte is specified when orb and other gene products concentrate in a single cell in region 2b of the germarium. A partially functional orb 3′UTR replacement delays oocyte specification until the egg chambers reach stage 2 of oogenesis. Before this point, orb mRNA and protein are unlocalized, as are other markers of oocyte identity, and the oocyte is not specified. After stage 2, ∼50% of the chambers successfully localize orb in a single cell, and this cell assumes oocyte identity. In the remaining chambers, the orb autoregulatory loop is not activated and no oocyte is formed. Finally, maintenance of oocyte identity requires continuous orb activity.
Authentic Representation and Author Identity: Exploring Mental Illness in The Hobbit Fanfiction
Authors: Rogers Jennifer
Published: October 2019

This paper addresses concerns with authenticity claims that surround mental illness and author identity in fanfiction. I will apply the critiques surrounding representation in media (see Mitchell and Snyder, 2001; Couser 2003, 2009) found in disability studies and fandom studies (see Jenkins 2012) to fanfiction. In this paper, I analyze two pieces of fanfiction which focus on Thorin II also known as Thorin Oakenshield, a character from J.R.R Tolkien’s The Hobbit novel and Peter Jackson’s film adaptations. I explore how in these texts the authors portray mental illness through their characterization of Thorin II. How the author’s actual or perceived personal mental health status may impact their writing, and readers’ responses to their writing, is explored through the lenses of identity politics (see Calhoun, 1994) and authenticity (Couser, 2009; van Dijk, 1989). In the context of disability and fandom studies, these fanfictions act as examples of a) combination fictional/ autobiographical writings which work to provide what the authors’ perceive as accurate portrayals of mental illness, and b) how the author’s mental health status impacts the perceived credibility of their work.
Normalizing Disability: Tagging and Disability Identity Construction through Marvel Cinematic Universe Fanfiction
Authors: Raw Adrienne E.
Published: October 2019

The exploration of identity is a common practice in fanfiction, and scholarship has consistently investigated this fan practice. Yet, despite the presence of disability and disabled characters in fanfiction, this aspect of identity exploration is only sparsely represented in scholarship. This article explores the intersection of disability studies and fanfiction studies through the lens of labelling and tagging, key elements of both fields. Labelling and classification in disability communities are often associated with medicalization, stereotyping, and erasure of individuality, while tagging in fanfiction provides a communicative framework between authors and readers. These differences in functions of labelling and tagging provide the foundation that enables tagging in fanfiction to function inclusively as a normalizing force, despite the problematic role of labelling in disability communities. Three trends in the ways disability is tagged in fanfiction are explored through a close reading of a selection of fanfiction from the Marvel Cinematic Universe: (1) disability is primarily tagged when it is a significant component of the plot, (2) the disability of canonically disabled characters is primarily tagged when that disability directly influences the plot of the story, and (3) mental disability/illness is significantly more represented than physical disability.
Seeking a Second Opinion on Social Media
Authors: Caruso Brown Amy E. , Arthur Joshua D. , Mutrie Lauren Hall , Lantos John D.
Published: October 2019

Social media pervades all aspects of our lives. In medicine, it has changed the ways that patients and parents get health information, advocate for particular treatments for themselves and their children, and raise money for expensive treatments. In this Ethics Rounds, we present a case in which the use of social media seemed to cross the boundaries of acceptable professionalism. What should the ground rules be for doctors who are tempted to give medical opinions online about patients whom they have never seen?
Penentuan Akar-Akar Sistem Persamaan Tak Linier dengan Kombinasi Differential Evolution dan Clustering
Authors: Badriyah Jamaliatul
Published: October 2019

Pada artikel ini akan dibahas tentang modifikasi dari Differential Evolution untuk menentukan akar-akar dari suatu sistem persamaan tak linier. Algoritma Differential Evolution ini nantinya akan dikombinasikan dengan teknik clustering. Algoritma Differential Evolution dengan clustering ini mampu untuk menentukan akar-akar dari system persamaan tak linier dalam satu kali running.
Model Autoregressive dengan Pendekatan Conditional Maximum Likelihood Untuk Prediksi Harga Saham
Authors: Rahmadayanti Cipta , Rabbani Hasbi , Rohmawati Aniq Atiqi
Published: October 2019

Jual beli saham merupakan salah satu bentuk investasi yang menjanjikan para investor, investasi berkaitan dengan return atau keuntungan yang didapatkan oleh suatu investor atas suatu investasi yang dilakukan terhadap saham tertentu. Untuk mendapatkan nilai return pada beberapa periode kedepan dapat dilakukan prediksi, pada dasarnya prediksi dapat dilakukan dengan menggunakan beberapa metode, namun dengan menggunakan model time series diharapkan menghasilkan prediksi yang baik karna karakteristik dari data saham merupakan data time series yang bergerak kontinu terhadap waktu. Pada penelitian ini digunakan model time series Autoregressive (AR) dengan pendekatan Conditional Maximum Likelihood untuk memprediksi nilai return serta dapat melihat pergerakan harga saham. Nilai parameter yang penting pada model Autoregressive orde 1 adalah . Hasil penaksiran parameter dengan Conditional Maximum Likelihood digunakan untuk memperoleh nilai hasil prediksi. Berdasarkan hasil analisis,  model Autoregressive dengan pendekatan Conditional Maximum Likelihood adalah model yang baik untuk memprediksi return dan harga saham NASDAQ dengan RMSE sebesar 0,0002578. Berdasarkan hasil prediksi model AR(1), maka para investor dapat membuat strategi untuk berinvestasi pada indek saham NASDAQ agar dapat menghasilkan keuntungan.
Peramalan Harga Saham Kioson di Era Ekonomi Digital
Authors: Windasari Wahyuni
Published: October 2019

Perkembangan teknologi dan perubahan ke era digital selain mempermudah investor untuk mendapatkan informasi mengenai suatu emiten di pasar modal juga berpengaruh terhadap keputusan investor untuk menanamkan sahamnya pada suatu emiten. Pesatnya kemajuan teknologi berimbas pada sentimen investor dalam memilih saham yang bergerak di bidang teknologi digital. Kioson merupakan salah satu perusahaan yang bergerak di sektor perdagangan online (e-commerce) dan teknologi yang mencatatkan sahamnya di Bursa Efek Indonesia (BEI). Ramainya investor yang berinvestasi pada saham KIOSON terjermin pada lonjakan pergerakan harga saham. Dalam penelitian ini, akan dilakukan peramalan harga saham Kioson dengan menggunakan metode ARIMA. Dari hasil penelitian menunjukkan model ARIMA AR(2) merupakan model terbaik untuk menggambarkan pergerakan harga saham kioson, dengan nilai MSE untuk data fittingnya sebesar 210,705.
Akurasi Peramalan Long Horizon dengan Singular Spectrum Analysis
Authors: Sakinah Awit Marwati
Published: October 2019

Perubahan iklim akhir-akhir ini tidak dapat dihindari. Salah satu penyebab perubahan iklim adalah perubahan suhu udara. Untuk itu, perlu dilakukan peramalan suhu agar penyimpangannya dapat diantisipasi. Dalam penelitian ini akan dibandingkan akurasi hasil peramalan dengan menggunakan model Singular Spectrum Analysis (SSA) dengan R-forecasting dan V-Forecasting. Peramalan dengan metode SSA R-forecasting dan V-Forecasting pada suhu Jakarta menggunakan window length L= 204 dan r=3 menghasilkan ramalan yang tidak jauh berbeda (aproksimasi kekontinuannya hampir sama). Berdasarkan hasil analisis, didapat MAPE untuk hasil permalan dengan SSA R-forecasting sebesar 5,0029 yang lebih besar dari MAPE SSA V-Forecasting sebesar 4,0067. Ini munjukkan bahwa peramalan suhu untuk long horizon lebih akurat dengan menggunakan V-Forecasting dibandingkan dengan R-Forecasting.
Storage Stability of Fish Waste Peptone at Ambient Temperature
Authors: Poernomo Achmad , Ariyani Farida , Murdinah Murdinah
Published: October 2019

Peptones from fish waste has been widely studied, however information about its shelf life is stilllimited. This study aims to test the storability of dried peptone from tuna and shrimp waste produced through hydrolysis using alcalase enzyme. Peptone powders were packed in HDPE plastic bottles and plastic coated aluminum foil, stored at room temperature, and periodically observed in quality (moisture content, aw, color and appearance). A test was also performed on their ability to support the growth of Staphylococcus aureusbacteria; all were compared to commercial peptone (Difco). Shrimp waste peptone had the highest moisture, ash calcium contents, while tuna peptone has the highest fat content. During five month storage at ambient temperature, all peptones experienced a slight decrease in quality. Aluminum foil performed better than HDPE bottles as a packaging material for peptones, i.e., able to maintain the moisture content, water activity, and appearance. Although the ability to support bacterial growth after five months of storage was slightly affected, the tested peptones were still able to beused as bacterial growing media. It can be concluded that fish waste peptones had comparable quality and shelf-life atambient temperature to commercial peptone.
Toehold switch assembly v1
Authors: Qian Junyan
Published: October 2019

General toehold assemble method using NEB Q5 High-Fidelity 2x Master Mix. Here we took Boir Noir toeholds as an example, it has four candidates we wanted to test. In order to have a higher yield, run 5 reactions for each candidates.
Silver Nanoparticles as a New Generation of Antimicrobial Prophylaxis
Authors: Garipov Ilnur T. , Khaydarov Renat R. , Gapurova Olga U. , Khaydarov Rashid A. , Lutfi Firdaus M. , Efimova, Irina L. , Evgrafova Svetlana Yu.
Published: October 2019

Over the last 20 years, metallic silver in the form of Ag nanoparticles has made a remarkable comeback as an example of a nanomaterial for control of microorganisms. The purpose of our study was a) to quantitatively estimate the antimicrobial effect of silver nanoparticles compared with that of silver ions and b) to check the efficacy of nanosilver as an antimicrobial agent against a range of microbes on the surface of water-soluble paint, 100% cotton fabric, and fibrous chemisorbent. Minimum inhibitory concentration tests quantitatively showed that Ag nanoparticles were less efficient than Ag+ ions against representatives of gram-positive / gram-negative bacteria and cosmopolitan saprotrophic fungi. Antifungal/antibacterial effects against Aspergillus niger, Penicillium phoeniceum, and Staphylococcus aureus were confirmed for nanosilver concentrations of even 1 μg/cm2 on the surface of cotton fabric and 0.8 μg/cm2 in water-soluble paint. As the concentration of nanosilver in water-soluble paint/cotton fabric was increased to 7 μg/cm2, the growth of Bacillus subtilis and Escherichia coli was suppressed as well. Microbiological tests conducted over a period of 60 days showed that there was no biofilm growth on the surface of a silver nanoparticle-coated fiber sorbent during its everyday operation as a household water treatment filter. Thus, silver nanoparticles as an add-on to water-soluble paints, textile fabrics or fiber chemisorbents had a remarkable antibacterial/antifungal effect, although some of the Ag nanoparticles were agglomerated into larger colloidal clusters
Nanotechnology-Based Delivery Systems: Highlights in Agricultural Applications
Authors: de Oliveira Jhones L.: , Campos Estefânia V.R. , Camara Marcela C. , de Carvalho Lucas B. , Monteiro Renata A. , de Lima Renata , Fraceto Leonardo F.
Published: October 2019

Due to the excessive use of chemical agents in agriculture, numerous problems have arisen, such as contamination of the environment, intoxication of non-target organisms and the development of resistance mechanisms by pests. To overcome these challenges, several sustainable technological approaches are being explored, and nanotechnology is one of them. This review aims to provide insights into the use of nanotechnology related to the agricultural sector. Articles were selected using the Web of Science and Science Direct databases; more than 50 manuscripts between 2015 and 2019 were reviewed. This review includes systems based on nanotechnology, in particular, for the sustained release of active ingredients for pest control, nutrition and plant growth. Nanoparticle-based formulations have great potential to increase agricultural productivity and reduce health and environmental impacts. However, there are certain technological challenges that must be addressed to allow the adoption of this technology for wider use in agri-food production
Closure of Earth’s Biosphere: Evolution and Current State
Authors: Bartsev Sergey I. , Degermendzhi Andrey G. , Sarangova Antonina B.
Published: October 2019

The existence of the biosphere is determined by the presence of a constant circulation of substances, carried out by a highly branched trophic network of mainly closed material loops. How this largely self-contained system formed remains unclear. The theory of evolution cannot help answer this question since the closure of the biosphere is not an adaptive trait of an individual – this is the essence of the Vernadsky-Darwin paradox. The present paper discusses stages of the formation of the biosphere in the context of closure – a key property and parameter of the biosphere – and possible approaches to resolving the paradox. The authors assume that the appearance of the first living organisms did not mean the appearance of the biosphere as a system of interacting components. The formation of the biosphere in the true sense of the word was associated with the appearance of predation approximately 500 million years ago and the emergence of a highly branched trophic network. The authors obtain simple estimates showing that, on the one hand, living organisms are potentially capable of changing their environment at the global level in a negligible geological time period but, on the other hand, are capable of maintaining an accurate balance of global material cycling for several tens of thousands of years. A simple model was used to show the effect of stoichiometric constraints on the formation of closed material flow in simple ecosystems and to demonstrate the need for increased diversity at trophic levels to overcome these stoichiometric constraints
Seismicity Monitoring in North-Central New Mexico by the Los Alamos Seismic Network
Authors: House Leigh , Roberts Peter
Published: October 2019

The first seismic stations of the Los Alamos Seismic Network (LASN) were installed in 1973, as a part of research on monitoring of nuclear testing. The extent of the network rapidly expanded by the late 1970s. By the middle 1980s, spatial coverage of the network was drastically reduced, due to a loss of funding. Since then, however, it has been possible to expand the coverage of the network with additional stations and to slowly upgrade network equipment to match contemporary instrument standards. These improvements will make it possible to keep recording the network’s data and to initiate real-time exchange of data with other institutions to improve earthquake monitoring throughout New Mexico and neighboring states. During more than 40 yr of operation, the network has provided a slow but steady increase in the volume of earthquake seismograms available to study seismicity and tectonics in north-central New Mexico. The current network covers an area from the Valles Caldera on the west, to within the Rio Grande rift on the east. LASN has yielded locations for about 900 earthquakes in north-central New Mexico between 1973 and 2013 (the most recent locations available). Epicenters of these have a complex pattern, with some that can be attributed to the deformation of the rift, though most are spread in areas west of the rift. A lack of seismicity in and near the Valles Caldera reinforces an earlier observation, based on far fewer earthquakes, which first called attention to this paucity of seismicity and attributed it to ductile deformation resulting from elevated crustal temperatures.
Screening transposon-mutant library of methicillin-resistant Staphylococcus aureus for mutants with altered sensitivity toward three antimicrobial agents
Authors: Ahmed Mahmoud , Beshara George , Ubanwa Bryan , Bae Taeok
Published: October 2019

Background and Hypothesis: Methicillin-resistant Staphylococcus aureus (MRSA) is resistant to almost all beta-lactam antibiotics, including oxacillin. To combat MRSA infections, recently, new classes of drugs are developed, such as oxadiazole DR1 and ESP404. We hypothesize that, if we identify MRSA mutants with altered sensitivity toward the drugs, those mutants will help us understand the antimicrobial mechanisms of the drugs.
Experimental Design or Project Methods: The screening was carried out on the Nebraska library consisting of 1,920 non-redundant transposon mutants of USA300 strain in 96 well plates. First, the minimum inhibitory concentration (MIC) of each drug was determined. Then the transposon mutants were grown in tryptic soy broth (TSB) containing either 1/4× MIC or 4× MIC of the drugs for sensitive mutants and resistant mutants respectively. Due to the limited availability of ESP404, the drug was used to screen for sensitive mutants only.
Results: The first screening identified 81 mutants with altered drug sensitivity: 23 for oxadiazole DR1 (5 sensitive, 18 resistant); 20 for ESP404 (all sensitive); 38 for oxacillin (18 sensitive, 5 resistant).
Conclusion and Potential Impact: Screening of the transposon-mutant library identified multiple mutants that have altered sensitivity toward three drugs against S. aureus. Further verification of the phenotype and characterization of those mutants are expected to shed light on the antimicrobial mechanism of the drugs and open a door for the development of drug-potentiators.
Predictors of Successful Outcomes Following Revision Total Knee Arthroplasty for Flexion Instability
Authors: Arnold Payton K. , Madden Joseph A. , Ziemba-Davis Mary , Meneghini, MD R. Michael
Published: October 2019

Background and Hypothesis: Flexion instability is a poorly understood cause of total knee arthroplasty (TKA) failure, often referred to as a “wastebasket” diagnosis for unexplained persistent pain. It remains a challenging diagnosis, with unpredictable outcomes following revision surgery. This study identified predictors of successful patient reported outcome measures (PROMS) following revision for flexion instability.
Methods: 121 consecutive revision TKAs for flexion instability from 2011-2018 at a single center were retrospectively reviewed. PROMS were prospectively obtained preoperatively and at minimum one-year follow-up. 33 potential predictors of PROMS encompassed: presenting symptoms, demographics/medical covariates, intraoperative observations, and pre/postoperative radiographic measurements. Variables related to outcomes with p≤0.20 in univariate analysis were entered in regression models. Nonsignificant predictors were pruned until only predictors with p<0.05 and the best model statistics were achieved.
Results: The sample was 63% female with mean age of 65±11 and BMI of 34±7 kg/m2. Pre- to postoperative changes in three radiographic measurements vital to the stability and function of TKA were related to function (p<0.001), pain (p=0.019), and satisfaction (p=0.030) after surgery. Pain with walking (p≤0.059) and stair climbing (p≤0.048) were almost exclusively related to demographic/medical covariates. Only one of the presenting symptoms commonly associated with flexion instability influenced early revision outcomes (p=0.021).
Conclusion and Potential Impact: Instability is one of the most common reasons for TKA failure leading to costly revision surgery. To our knowledge, this is the largest flexion instability cohort studied to date, including substantially more predictors in an attempt to elucidate factors indicative of successful treatment outcomes. With the exception of radiographic parameters reflecting surgical decisions, findings suggest that many factors related to outcomes may be beyond the control of arthroplasty surgeons.
Strategies to Increase Time in Range in Children Wearing Continuous Glucose Monitors
Authors: Bridgeman Will , DiMeglio Linda
Published: October 2019

Background and Hypothesis: Optimal management of Type 1 Diabetes (T1D) requires careful orchestration of insulin dosing with blood glucose values, food intake, activity levels, and concurrent illness. This is particularly burdensome for caretakers of very young children. Continuous glucose monitors (CGMs) provide useful real-time blood glucose information. Yet, data suggest that their use in young children can reduce severe hypoglycemia but does not consistently improve the time that blood glucose values are in an optimal range.
Project Methods: We first analyzed survey data from a study of CGM use in 143 children 2-8 years of age). We then developed a semi-structured qualitative interview, and queried parents of children using CGM under 9 years of age (mean age 5.2±1.6 years, A1c 7.8±0.9%).
Results: Recurrent themes identified included: CGMs and remote monitoring brought parents peace of mind and permitted more flexibility in care. Trend arrows indicating rapid glucose decreases induced “anxiety” and “panic” in caretakers, due to fear of hypoglycemia and immediate safety. Rising trend arrows induced frustration, but were less likely to change management decisions, due to lack of immediacy of risk of long-term complications. CGM data can be “overwhelming”, but third party apps and “experience” increase helpfulness.
Conclusion/Potential Impact: Our initial data suggest that behavioral/educational interventions to improve time in range must incorporate implementable strategies, potentially using apps, to encourage parents to respond to and reduce time spent in higher blood glucose ranges.
Cell cycle analysis of MDA-MB-157 and APC knockdown cells
Authors: Brown Jamie , Astarita Emily , Hoover Camden , Prosperi Jenifer R.
Published: October 2019

Background and Hypothesis: A majority of sporadic breast cancers include deficits in the expression of Adenomatous Polyposis Coli (APC), a tumor suppressor. Deficits in APC are more common in patients with TNBC (triple negative breast cancer), which is also a cancer prone to chemotherapy resistance. The Prosperi lab previously found that APC knockdown cells (APCKD) were resistant to Paclitaxel (PTX). We hypothesize that APCKD cells are resistant to PTX treatment through avoidance of G2/M arrest. This summer, my goal was to investigate the mechanism by which PTX works to arrest the cell cycle at G2/M. 
Experimental Design or Project Methods:
Cell Synchronization: To synchronize MDA-MB-157 and APCKD cells, we first tried serum starvation for 24-72 hours. Second, we tried synchronizing the cells using a double thymidine block as described (Bostock, 1971). In either protocol, cells were then stained with Propidium Iodide (PI) and flow cytometry was performed. Paclitaxel (PTX) treatment and cell cycle analysis: MDA-MB-157 and APCKD cells were grown to confluency and then treated with 0.078µM PTX for 12, 24, and 48 hours. Cells were stained with PI and flow cytometry was performed.
Results:
Cell synchronization: APCKD cells cells have an increased cell population in the G2/M phase than the parental cells after serum starvation. Importantly, APC knockdown cells are not impacted by serum starvation up to 72 hours. Additionally, a double thymidine block is insufficient to synchronize MDA-MB-157 and APCKD cells. A double thymidine block did shorten the S phase and move MDA-MB-157 and APCKD cells closer to G0-G1 arrest, but did not synchronize. Paclitaxel (PTX) treatment and cell cycle analysis: MDA-MB-157 and APCKD cells treated for longer intervals experienced more cell death and were further arrested in G2/M.
Conclusion and Potential Impact: We learned that MDA-MB-157 and APCKD cannot be easily synchronized using serum starving or a double thymidine block. Future investigations will require alternative methods of synchronization or will proceed without synchronization. Furthermore, APCKD cells do not avoid G2/M arrest when treated with Paclitaxel, indicating a different mechanism of PTX resistance.
Asthma in Indiana – Using a Community Health Matrix to Determine Asthma Health Factors for Indiana Counties
Authors: Burrell Mikayla , Casey, MPH Rachael , Savaiano, PhD Dennis
Published: October 2019

Background and Hypothesis: Asthma and its appropriate treatment is a public health issue in Indiana that Indiana Joint Asthma Coalition (InJAC), a partnership within CTSI, is attempting to address. This is done through state-wide coalition building, which unifies efforts regarding asthma health and education and promotes interprofessional collaboration. Because time and resources are limited, InJAC must choose the areas that would benefit most from their focused work. A matrix was developed to establish the 10 counties with poorest asthma health and high vulnerability to social determinants to aid in this choice. We hypothesize that the 10 counties with the highest vulnerability to social determinants of health will have the worst asthma health.
Project Methods: Asthma health outcomes, contributing asthma-related variables, and social determinants of health were identified in all 92 counties in Indiana. Counties were compared by composite z scores to determine the top 10 counties with the poorest health statistics for asthma and social determinants. In addition, qualitative data will be used to identify local health coalitions that have the capacity and desire to work with InJAC to improve asthma treatment. InJAC will begin sessions with these counties to determine if long-term, sustainable, health promotions are feasible.
Results: The top 10 counites that were identified as having the poorest asthma health and factors were Lake, Grant, Madison, Marion, Huntington, Vanderburgh, Howard, La Porte, Blackford, and Noble. The top 10 counties with highest vulnerability to social determinants were Owen, Ripley, Daviess, La Grange, Fayette, Wayne, Elkhart, Newton, Switzerland, and Marion.
Potential Impact: The data from this matrix will help direct InJAC to the areas of Indiana with the most need for asthma coalition efforts. This will be done through improvement on education, awareness, and quality of care based on the Indiana State Asthma Plan.
Xenograft Bone-Derived Collagen Scaffold for Myelomeningocele Management
Authors: Khan Sadid , Campbell Natalie , Fathizadeh Sam , Bucherl Sean , Nauman, Ph.D. Eric
Published: October 2019

Background and Hypothesis: Spina bifida is a neural tube defect resulting from an incomplete closure of the caudal neuropore. The most debilitating form of spina bifida, myelomeningocele (MMC), can present with Chiari II malformation with concomitant hydrocephalus, bowel and bladder abnormalities, and impaired motor function of the lower limbs. The incidence rate of spina bifida is 3.4 per 10,000 live births reported within the US. On average, the US spends $1,176,000,000 annually on patient management and treatment. Advancements in existing treatment options, namely fetal surgery, can greatly decrease neurological injury and related costs, but at the risk of fetal and maternal complications. Various tissue engineering methods have been proposed including biodegradable and synthetic scaffolds, seeded with or without bioactive proteins and stem cells, sutured or glued to the defect, and administered fetoscopically or through open fetal surgery. However, no combination of these methods is fully biointegratable, watertight, provides complete coverage with adequate mechanical strength, and is able to be administered fetoscopically.
Experimental Design: This study utilizes bovine and porcine bone to create an organic, flexible collagen scaffold that can be seeded with bioactive proteins and attached with adhesive for successful coverage of MMC defects.
Conclusion and Potential Impact: The natural matrix may allow for quicker host cell integration and greater mechanical strength compared to existing models. This study will characterize the mechanical strength, permeability, and biointegration of the proposed management of spina bifida.
Obesity decreases the contribution of Kv channels to hypoxic coronary vasodilation
Authors: Clark Hannah E. , Baker Hana E. , Goodwill Adam G. , Blaettner Bianca S. , Kozlowski Michael C. , Tune Johnathan D.
Published: October 2019

Background and Hypothesis: Our group previously demonstrated that reductions in the functional expression of voltage-dependent K+ (Kv) channels contribute to impaired metabolic control of coronary blood flow in the setting of obesity. This study tested the hypothesis that obesity diminishes the contribution of Kv channels to coronary vasodilation in response to hypoxemia.
Experimental Design or Project Methods: Control lean (n = 7) and obese (n = 5) swine were anesthetized and the heart exposed via left lateral thoracotomy. Coronary blood flow was measured in response to hypoxemia, before and after inhibition of Kv channels by 4-aminopyridine (4-AP; 0.3 mg/kg, iv), by a flow probe placed about the left anterior descending coronary artery. Hypoxemia was induced by progressive increases in the amount of nitrogen introduced into the ventilator. Arterial blood samples were obtained at each reduction in arterial oxygenation via a catheter placed in the femoral artery.
Results: Blood pressure decreased from ~88 ± 5 mmHg to ~68 ± 6 mmHg (P = 0.01) as arterial PO2 was reduced below 50 mmHg in both lean and obese swine (P = 0.51). In lean swine, coronary flow progressively increased from ~0.6 to >3.0 ml/min/g as arterial PO2 was reduced. This response was decreased by ~40% in obese swine and by ~30% in lean swine treated with 4-AP. Administration of 4AP had no effect on coronary flow in obese swine.
Conclusion and Potential Impact: These data support that Kv channels contribute to increases in coronary flow in response to hypoxemia in lean swine and that reductions in Kv channel function contribute to impaired hypoxic coronary vasodilation in obese swine. We propose that therapeutic targeting of obesity associated pathways (angiotensin-aldosterone system) known to influence K+ channel expression could improve coronary microvascular function and cardiovascular outcomes in subjects with obesity. Supported by R01 HL136386; T35 HL 110854.
Defining the expression and function of COQ8B in TAA
Authors: Cuevas Christian C. , Ware Stephanie M. , Landis Benjamin J.
Published: October 2019

Background and Hypothesis: Thoracic aortic aneurysm (TAA) is characterized by progressive enlargement of the aorta and predisposes to life-threatening aortic dissection. Even among patients with monogenic causes of TAA, variable expressivity and incomplete penetrance hinders clinical decisions. Recent studies identify the gene COQ8B, a key mediator of mitochondrial ubiquinone biosynthesis, as a candidate genetic modifier in TAA. First, we sought to describe the distribution and morphological appearance of mitochondria in aortic smooth muscle cells (SMCs). We further hypothesized that decreased expression of COQ8B alters the contractile homeostasis of SMCs and predisposes to SMC apoptosis.
Experimental Design or Project Methods: We cultured SMCs directly from aortic tissues acquired at the time of aortic replacement surgery or heart transplantation. Mitochondria were labeled in live and fixed SMCs using the MitoTracker dye. We knocked down expression of COQ8B in SMCs using siRNA (siCOQ8B). The effects of siCOQ8B were compared with SMCs treated with a non-targeting negative control siRNA (siNeg). RNA expression was measured using quantitative polymerase chain reaction (qPCR) at 48 to 72 hours after siRNA transfection. Apoptosis was measured using a TUNEL-based fluorescent assay.
Results: Mitochondria in aortic SMCs showed a predominantly perinuclear distribution along with the appearance of a network of thin, filamentous extensions outward from the nucleus. Compared with siNeg controls, siCOQ8B decreased COQ8B RNA levels by approximately 90% on average across 5 knockdown experiments (80 to 95% knockdown). siCOQ8B led to increased RNA levels of CNN1 across all experiments, ranging from 1.4 to 7.4-fold increases. Meanwhile, siCOQ8b led to lower levels of COL1A1 in 4 of 5 experiments, ranging from 1.1 to 1.9-fold decreases. siCOQ8B led to decrease in the percentage of apoptotic cells in comparison with siNeg. In the presence of 1.6mM of H2O2, there was a 15% increase in apoptosis in cells transfected with siCOQ8B, and a 4.5% increase in cells not treated with H2O2.
Conclusion and Potential Impact: Decreased expression of COQ8B via siRNA knockdown alters the expression of genes important for SMC homeostasis, causing the SMC to be more susceptible to oxidative stress and inducing apoptosis. Future studies will further explore the role of COQ8B on SMC contractility, matrix production, and apoptosis and elucidate the underlying mechanism. Ultimately, this insight may lead to novel clinical approaches and therapies for patients with TAA.
Buprenorphine Metabolism in Human Liver, Human Placenta, and Recombinant Cyp19 Microsomes
Authors: Enderle Kaitlyn , Zheng Xiaomei , Quinney,  PharmD, PhD Sara
Published: October 2019

Background and Hypothesis: Buprenorphine (BUP), a partial opioid agonist, is commonly used to treat opioid use disorder in pregnant women. BUP is metabolized in the liver by cytochrome P450s (CYP) and UDPGlucuronosyltransferases (UGT). Compared to non-pregnant women, higher doses of BUP are needed during pregnancy to maintain therapeutic concentrations. We hypothesize that the placenta has a role in the metabolism of BUP, thus contributing to the need for higher doses during pregnancy.
Experimental Design or Project Methods: : BUP was incubated with human liver microsome (HLM), human placenta microsome (HPM) or recombinant CYP19 microsome (rCYP19). Norbuprenorphine (Nor-BUP) formation was measured by HPLC/MS/MS (Thermo TSQ Quantum Ultra). Positive controls included midazolam for HLMs and testosterone for HPMs and rCYP19, and negative controls were without NADPH. Initial experiments assessed linearity with time and protein concentration. Subsequently, BUP concentrations were varied and Nor-BUP formation vs. BUP concentration was fitted to the Michaelis–Menten equation (GraphPad Prism) to calculate Km and Vmax.
Results: The Km was 11.89 μM and Vmax was 0.4627 μmol/min/mg for Nor-BUP formation following incubation of BUP (0-20 µM) with HLM. Higher concentrations of BUP indicated non-typical Michaelis–Menten kinetics. Results from the HPM show metabolism of BUP to Nor-BUP is present in the placenta. Incubation of BUP in rCYP19, an enzyme known to be in human placenta, resulted in Nor-BUP formation, indicating its role in BUP metabolism.
Conclusions and Potential Impact: : The Km and Vmax values generated from concentrations of 5-20 μM in HLM is comparable with existing data. Because multiple enzymes breakdown BUP with different Km and Vmax values, a curve not adhering to the normal Michaelis–Menten shape over 20 μM BUP is reasonable in HLMs. Formation of Nor-BUP following incubation of BUP in rCYP19 and HPMs suggest their role in the metabolism of BUP in pregnant women. Understanding the role of placenta in metabolizing BUP will provide insight to fetal drug exposure. Placental metabolism of BUP would possibly indicate lower exposure to BUP but higher exposure to nor-BUP to the fetus. Future directions of this study will quantify placental metabolism of BUP, investigate other pathways of BUP metabolism in the placenta, and determine drug interactions that may exacerbate Neonatal Abstinence Syndrome when coadministered with BUP.
Long Term Outcomes of Iliac Artery Aneurysm (IAA) and Abdominal Aortic Aneurysm (AAA) Repair with Endovascular Procedures at the Parkview Heart Institute
Authors: Gray, M.D. Douglas , Herrera, B.S. Christopher , Recht, N.P. Kim
Published: October 2019

Background: Favorable characteristics of the minimally invasive endovascular aneurysm repair (EVAR), such as a reduced recovery time, have led physicians and patients alike to elect for this procedure over open repair. Longitudinal study on endovascular repair has raised concerns regarding an increased rate of late failure leading to rupture and overall higher rates of re-intervention for patients.
Objective: This project compares the outcomes of endovascular stent graft repairs of AAA and IAA at Parkview Heart Institute over a certain time span.
Methods & Design: This study is a retrospective review of patients (N=89) with AAA/IAA repair since 2014. The EPIC online medical record is the main tool in data extraction while statistical analysis was conducted via Microsoft Excel (p = 0.05).
Results: Males, ages 51-75 y.o., White population, non-Hispanics, and BMI rating of 1-30 were observed to have more AAAs. A significant decrease in length of stay over historical averages was observed with at most 1-3 days and minimal complications. Most causes for re-interventions were due to endo-leaks. More than 60% of patients with AAA and EVAR have a significant past medical history of smoking, hypertension, and chronic obstructive pulmonary disease. Finally, most pre- and post-operative aneurysms sizes are within 41-60 mm in diameter.
Conclusion and Potential Impact: This study analyzed 89 patients that experienced AAA instead of IAA. Certain populations, such as being male, ages 51-75 y.o., White population, non-Hispanics, and BMI rating of 1-30 with past medical history characteristics are frequent in patients with EVAR. The procedure has minimal complications in comparison to open abdominal surgery, and this study will add to the literature by aiding physicians’ determination of the best course of action for treating patients with AAA/IAA.
Neonatal bowel resections occurring in the NICU are associated with lower survival rates compared to the OR
Authors: Howser Lauren , Goehring Carly , Anderson Cassandra , Turk Eamaan , Han Yan , Eckert George , Gray, MD Dr. Brian
Published: October 2019

Background and Hypothesis: Neonatal surgical procedures frequently occur at bedside in the neonatal intensive care unit (NICU) rather than in the operating room (OR). The goal of this study was to isolate risk factors for neonatal bowel resections occurring in the NICU, compare outcomes of bowel resections occurring both in the NICU and OR, and evaluate modifiable factors in both locations.
Experimental Design or Project Methods: We reviewed 139 neonatal (<30 days old) patients at Riley Hospital for Children who underwent bowel resection in either the NICU or the OR from 2014-2018. Statistical analysis included bivariate, multivariable, and propensity score matching analyses.
Results: When compared to neonates who had bowel resection in the OR, the NICU group had lower mean gestational age (EGA) at birth (26 vs 34 weeks, p=<0.0001), lower weight at surgery (72.7% NICU patients <1000 grams vs 7.6% OR patients, p=<0.0001), and had the operative diagnosis of spontaneous intestinal perforation (48.8%) or NEC (42.9%) compared to OR patients (18.7%) for both diagnoses (p=<0.0001, p=0.0003). Bivariate analysis revealed that EGA at birth (OR 0.668, p=<0.0001), APGAR 1 minute (OR 0.776, p=0.001), and pre-op respiratory FiO2 percentage (OR 2.514, p=<0.0001) were significant predictors of operation location. Propensity score matching (15 matched pairs) showed survival rates were lower in the NICU group (p=0.0156). Modifiable risk factors, such as the usage of a heating device during surgery (p=<0.0001) and temperature monitoring during surgery (p=<0.0001) were positively correlated with survival, while blood administration in the first 24 hours post-surgery (p=0.001) was correlated with worse survival.
Conclusions: There are definable characteristics of neonates who have bowel resections in the OR vs NICU, but the primary outcome difference between these groups is survival. We also identified modifiable factors that can be studied to further improve survival of patients with operations in both settings.
Impact of T Helper Cell Subtype on CAR-T Cell Therapy Efficacy
Authors: Khan, BS BA Ibrahim M. , Nelson, PhD Andrew S. , Kaplan, PhD Mark H.
Published: October 2019

Background and Hypothesis: Chimeric antigen receptors (CARs) are recombinant receptors with high affinity for the target antigen. Used for tumor therapy, CARs are transduced into patient T cells. CAR-T cells specific for CD19 are used to treat B cell acute lymphoblastic leukemia (B-ALL). Cancerous B cells are destroyed by CAR-T cells in an antigen-specific manner. Currently being used in conjunction with radiation and other cancer therapies to prohibit relapse, Dr. Marco Davila of the Moffitt Cancer Center, has shown that CAR-T therapy induces long term remission and B cell aplasia.
Experimental Design: In this experiment the CAR vector obtained from Dr. Davila was transduced into T helper cells cultured under varying conditions (Th0, Th9, and ThGranzyme A). B cell killing and longevity of transduced CAR-Th cells were monitored as part of the criteria for determining the most effective Th subtype for the CAR-T therapy. The target cell killing-mechanism was analyzed at the RNA level using quantitative polymerase chain reaction (qPCR) to analyze gene expression of cytotoxic molecules including granzymes A/B, perforins, Fas-FasL, and TNF-α. Th9 cells were expected to be among the most effective of the indicated subtypes due to their longevity and coordination of the immune response.
Results: T cells in all conditions were effectively transduced for CAR expression, although Th9 cells demonstrated a greater proportion of cultured cells that were transduced with the CAR. QPCR results suggest that there is specification of cytotoxic programs among the culture conditions. In Th9 cells, qPCR results suggest their use of perforin and TNF-α. Ongoing studies will compare cytotoxic activity.
Potential Impact: Further steps after determining the most effective culture conditions include injecting transduced Th cells of the optimized subtype into mice afflicted with BALL to assess cancer killing in vivo as well as the potential harm of the therapy to the patient.