Digital Commons Network^™

Tfeb-Vacuolar Atpase Signaling Regulates Lysosomal Function And Microglial Activation In Tauopathy, Baiping Wang, Heidi Martini-Stoica, Chuangye Qi, Tzu-Chiao Lu, Shuo Wang, Wen Xiong, Yanyan Qi, Yin Xu, Marco Sardiello, Hongjie Li, Hui Zheng

Center on Aging Staff Publications

Transcription factor EB (TFEB) mediates gene expression through binding to the coordinated lysosome expression and regulation (CLEAR) sequence. TFEB targets include subunits of the vacuolar ATPase (v-ATPase), which are essential for lysosome acidification. Single-nucleus RNA sequencing of wild-type and PS19 (Tau) transgenic mice expressing the P301S mutant tau identified three unique microglia subclusters in Tau mice that were associated with heightened lysosome and immune pathway genes. To explore the lysosome-immune relationship, we specifically disrupted the TFEB-v-ATPase signaling by creating a knock-in mouse line in which the CLEAR sequence of one of the v-ATPase subunits, Atp6v1h, was mutated. CLEAR mutant exhibited …

Health Disinformation-Gaining Strength, Becoming Infinite, Peter J Hotez

Faculty, Staff and Students Publications

No abstract provided.

Point Of Care Ultrasound (Pocus) For Advanced Practice Providers (Apps), Crystal S. Rogers

DNP Projects

Background: Advanced practice providers (APPs) routinely lack the skillset to perform Point of Care Ultrasound (POCUS) because currently, there is not a standardized curriculum within scholarly programs (Santos et al., 2018) and/or inadequate hospital training to perform these competencies (Huang et al., 2021). The absence of a standard curriculum has limited the ability of APPs to practice with full scope, provide care that is comparable to physician partners in the acute care setting, and deliver highest quality of patient care. Implementing standard education to train APPs for POCUS addresses this identified gap.

Purpose: The purpose of this project was to …

Autonomic Modulation Of Cardiac Function In Hypertrophic Cardiomyopathy, Jenna Rose Disser

Dissertations, Master's Theses and Master's Reports

Hypertrophic cardiomyopathy (HCM) is the most common genetic heart disorder. HCM is characterized by cardiac hypertrophy, fibrosis, and an increased risk of fatal arrhythmias and sudden cardiac death. Previous studies in humans with HCM have demonstrated increased cardiac norepinephrine spillover and reduced beta-1 receptor (β1R) expression in the left ventricle (LV). We have previously demonstrated that cardiac sympathetic tone is elevated in an alpha-tropomyosin mutant mouse model of HCM. We hypothesized that HCM mice would demonstrate reduced β1R expression in the LV and attenuated heart rate (HR) and dP/dt max responses to ramped infusion of the β1R agonist, dobutamine. …

Addressing Healthcare Disparities Faced By Aging Down Syndrome Populations, Rhea Mistry

Pitzer Senior Theses

Down syndrome (DS) is a chromosomal genetic condition that impacts the development of the body and brain. The life expectancy of individuals with DS has drastically improved over the decades, increasing from 9 years in 1929 to 25 years in 1983 and further reaching 60 years by 2002. Individuals with DS face a variety of prominent co-occurring health conditions that are subject to change over time, including cognitive, visual, and hearing impairments, sleep apnea, cardiovascular defects, Down syndrome regression disorder, and dementia. Despite this increase in life span, DS is still most familiar to pediatric care providers, resulting in healthcare …

Inflammation And Tumor Progression: The Differential Impact Of Saa In Breast Cancer Models, Daniel Wilhelm Olivier, Carla Eksteen, Manisha Du Plessis, Louis De Jager, Lize Engelbrecht, Nathaniel Wade Mcgregor, Preetha Shridas, Frederick C. De Beer, Willem J. S. De Villiers, Etheresia Pretorius, Anna-Mart Engelbrecht

Saha Cardiovascular Research Center Faculty Publications

Background: Previous research has shown that the Serum Amyloid A (SAA) protein family is intricately involved in inflammatory signaling and various disease pathologies. We have previously demonstrated that SAA is associated with increased colitis disease severity and the promotion of tumorigenesis. However, the specific role of SAA proteins in breast cancer pathology remains unclear. Therefore, we investigated the role of systemic SAA1 and SAA2 (SAA1/2) in a triple-negative breast cancer mouse model.

Methods: Syngeneic breast tumors were established in wild-type mice, and mice lacking the SAA1/2 (SAADKO). Subsequently, tumor volume was monitored, species survival determined, the inflammatory profiles of mice …

Impact Of Tiered Implementation Of Clinical Decision Support System For Acute Kidney Injury And Nephrotoxin Exposure In Cardiac Surgery Patients, Christopher M. Justice Ii

Graduate Theses, Dissertations, and Problem Reports (ETD)

Background: Nephrotoxin exposure may worsen kidney injury and impair kidney recovery if continued in patients with acute kidney injury (AKI).

Objective: To determine if tiered implementation of a clinical decision support system (CDSS) would reduce nephrotoxin use in cardiac surgery patients with AKI.

Methods: We assessed patients admitted to the cardiac surgery intensive care unit at a tertiary care center from January 2020 to December 2021, and August 2022 to September 2023. A passive electronic AKI alert was activated in Jul 2020, followed by an electronic nephrotoxin alert in March 2023. In this alert, active nephrotoxic medication orders resulted in …

Combinatorial Atoh1, Gfi1, Pou4f3, And Six1 Gene Transfer Induces Hair Cell Regeneration In The Flat Epithelium Of Mature Guinea Pigs, Yujie Liu, Lin Yang, Sunita Singh, Lisa A Beyer, Diane M Prieskorn, Donald L Swiderski, Andrew K Groves, Yehoash Raphael

Children’s Nutrition Research Center Staff Publications

Flat epithelium (FE) is a condition characterized by the loss of both hair cells (HCs) and supporting cells and the transformation of the organ of Corti into a simple flat or cuboidal epithelium, which can occur after severe cochlear insults. The transcription factors Gfi1, Atoh1, Pou4f3, and Six1 (GAPS) play key roles in HC differentiation and survival in normal ears. Previous work using a single transcription factor, Atoh1, to induce HC regeneration in mature ears in vivo usually produced very few cells and failed to produce HCs in severely damaged organs of Corti, especially those with FE. Studies in vitro …

Receipt Of Follow‑Up Care Plans On Colorectal Cancer Screening Among Breast, Prostate, And Lung Cancer Survivors, Meng-Han Tsai, Malcolm S. Bevel, Sydney E. Andrzejak, Justin X. Moore

Behavioral Science Faculty Publications

Purpose Our study aimed to examine whether receipt of follow-up care plans is associated with greater guideline-concordant CRC screening stratified by breast, prostate, and lung cancer survivors.

Methods We used data from years 2016, 2018, and 2020 of the Behavioral Risk Factor Surveillance System on 3339 eligible treatment-utilizing cancer survivors with complete treatment. We performed descriptive statistics and multivariable logistic regression to examine the mentioned association.

Results We observed that 83.9% of breast and 88.2% of prostate cancer survivors with follow-care plans received CRC screening (p-value < 0.001). The lowest CRC screening use was observed among lung cancer (70.8%). In multivariable analysis, receipt of follow-up care plans was strongly associated with greater odds of receiving CRC screening in breast (OR, 2.67; 95% CI: 1.71–4.16) and prostate (OR, 3.81; 95% CI: 2.30–6.31) cancer survivors. Regardless of provider type, 84 to 88% reduced likelihood of receipt of CRC screening when they received follow-up care plans among lung cancer survivors. Among those without follow-up care plans, breast (OR, 0.29; 95% CI: 0.09–0.92) and lung (OR, 0.05; 95% CI: 0.01–0.25) cancer survivors who received care from general practices were less likely to receive CRC screening compared to those who received care from non-general practices.

Conclusions Receipt of follow-up care plans was associated with greater CRC screening use in …

Us Hospices’ Approach To Racial/ Ethnic Minority Inclusion: A Qualitative Study, M. Courtney Hughes, Erin Vernon, Monica Kowalczyk, Marlon Basco-Rodillas

Faculty Articles, Papers, and Other Scholarship

Objective: To examine hospices’ approaches to improving the current racial/ethnic hospice utilisation disparity.

Methods: During June and July 2020, we conducted in-depth, semistructured interviews with 22 hospice leaders from across the USA. The interviews focused on inclusive strategy approaches. We analysed the data using qualitative methods.

Results: Multiple themes emerged about racial/ ethnic minority inclusion strategies: (1) hospices tailor strategies to the local communities they serve; (2) improvement involves addressing social determinants of health that extend beyond end-of-life care; (3) costs of strategies are not a primary concern and the benefits are worth the costs; and (4) hospices want to …

Role Of Magnetic Resonance Imaging (Mri) In Evaluating The Indeterminate Breast Lesions, Jehan Ibrahim M. Altohamy, Nivine Abdel Moneim Chalabi, Mohamed Amr Farouk, Mina Awny Moawad, Shrouk Mohamed Awadallah

Journal of Medicine in Scientific Research

Background: the number of women who are diagnosed with breast cancer (BC) continues to rise, making it the utmost public cause of cancer death among ladies globally. It has been proposed that high-risk individual's get breast MRIs, which are the most sensitive breast cancer screening method that does not use ionizing radiation. Aim and objectives; This research effort assesses breast MRI in order to clarify the Breast Imaging Reporting and Data System categorization of indeterminate mammography or ultrasonography examines as well as evaluate the diagnostic reliability of MRI in assessing lesions discovered on screening or diagnostic mammography or ultrasonography that …

Understanding The Effects Of Hormone Treatments On The Transgender Singer: A Pedagogical Study And Voice Studio Guide, Erin M. Hannon

Theses and Dissertations--Music

Transitioning from one gender to another can be an arduous and emotionally charged experience, accompanied by many physical and mental transformations. The journey of self-discovery and vocal explorations is both unique and deeply personal. Modifications that emerge in an individual's vocal characteristics have a profound influence on their capacity to communicate and express themselves.

Transgender vocal students and professional singers must navigate the delicate balance between their vocal and gender identities while considering the possible risks of Gender Affirming Hormone Therapy. Though these therapies may significantly alter one's appearance and overall well-being, they are also capable of causing notable declines …

Telemedicine And Physical Visits: A Dynamic Duo In Obesity Management Among Adolescents, Egypt, Enas Sayed Abbas, Safaa Al Hussein Tawfik, Gehan Fouad Ahmad, Lamiaa Ibrahim Haroon, Taghreed K. M. Elhady, Amira Said Nassar

Journal of Medicine in Scientific Research

Childhood obesity, a serious global issue, leads to various health problems like diabetes, high blood pressure, high cholesterol, and psychological issues. In Egypt, inconsistent data shows a significant prevalence of obesity among adolescents, influenced by factors like gender, age, and lifestyle. The study aims to explore the effectiveness of telemedicine, in conjunction with physical visits, for managing obesity among adolescents in Egypt. Telemedicine, a cost-effective and engaging alternative to in-person visits, can help patients lose weight and provide a safe, remote alternative, expanding treatment access to hard-to-reach populations. The study is a controlled clinical trial on overweight or obese adolescents …

Tim/Tam Receptors: A Potential Biomarker For Predicting Sensitivity To Zika Virus-Induced Oncolysis In Non-Small Cell Lung Cancers, Shankari Somasekar

Honors Undergraduate Theses

Non-small cell lung cancers (NSCLC) constitute 80-85% of lung cancers and are the leading cause of cancer-related deaths globally. The most common cause is prolonged smoking. Current treatment options for NSCLC include surgery, radiation, chemotherapy, targeted drug therapy, and immunotherapy. Although these medications are effective in the short term, patients often face issues of drug resistance and debilitating side effects with prolonged use. Currently, the use of Zika virus (ZIKV) is being researched as a possible alternative treatment for cancer, which minimizes side effects and the risk of drug resistance. TIM/TAM proteins are identified as the putative ZIKV receptors on …

Voices In End-Of-Life Care: Gathering Perspectives Of Speech-Language Pathologists On End-Of-Life Issues In Medical Settings, Kori Bess Benton

Online Theses and Dissertations

Despite healthcare professionals becoming increasingly aware of the needs of the end-of-life care population, little research exists regarding the roles speech-language pathologists play in providing care for that population. This study investigates perspectives of speech-language pathologists on their roles and preparedness for engaging in end-of-life care. Eight participants engaged in semi-structured interviews to share their experiences in end-of-life care. Interviews were conducted and transcribed, then analyzed to establish common themes amongst speech-language pathologists working in this setting. This study found that speech-language pathologists do not feel they received adequate training related to their role in end-of-life care and share unique …

Preferences And Feasibility Of Long-Acting Technologies For Treatment Of Hepatitis C Virus In Low- And Middle-Income Countries: A Survey Of Providers And Policymakers, Neil Gupta, Susan Swindells, Kimberly Scarsi, Renae Furl, David L. Thomas, Ethel D. Weld, Joelle Dountio Ofimboudem, Hailemichael Desalegn, Saeed Hamid, Alethse De La Torre Rosas, Angelica E. Miranda, Andrew Owen, Steve Rannard, Lindsey Hiebert, Katherine Sun, John W. Ward

Journal Articles: Infectious Diseases

Long-acting technologies (LATs) for hepatitis C virus (HCV) are under development as a strategy to improve linkage to care, treatment adherence and outcomes. We conducted a survey of HCV treatment prescribers and HCV policymakers in low- and middle-income countries (LMICs) regarding acceptability and feasibility of HCV LATs. We included one-time intramuscular injection, subdermal implant and transdermal patch as potential LAT options. We surveyed participants regarding optimal health system and patient characteristics, concerns, potential barriers, overall feasibility and preferences for HCV LAT as compared to daily oral medication. Overall, 122 providers and 50 policymakers from 42 LMICs completed the survey. Among …

Understanding Guardian Perceptions Of Whole-Foods Plant-Based Diets As Interventions In Pediatric Medicine, Vijay R. Luthra

Honors Undergraduate Theses

The obesity epidemic is widespread in American society, often affecting children who bear its consequences innocently. Due to a lack of nutrition knowledge, children rely on caregivers to meet their nutritional needs and instill healthy habits. This study examines parents' attitudes, knowledge, and behaviors, as well as perceived barriers to adopting a whole-foods plant-based diet (WFPD) for their children. Using a modified survey instrument by Morton et al., this study focused on parents and children regarding dietary choices. Participants were recruited from the waiting area of a pediatric clinic at the Medical College of Georgia between April and May 2023. …

Unraveling The Effects Of The Covid-19 Pandemic On Staphylococcus Aureus Carriage: A Comparative Research Of Pre- And Post-Pandemic Samples, Catherine G. Johnston

Honors Undergraduate Theses

The COVID-19 pandemic brought about significant changes in the epidemiology of Staphylococcus aureus carriage and disease. This study aims to assess these changes by leveraging respiratory samples collected from a longstanding cohort, comparing the genomic characteristics of S. aureus isolates from pre- and post-pandemic periods. We analyzed S. aureus genomes and phenotypically determined macrolide resistance to characterize population structure, antibiotic resistance, and virulence profiles. Changes in S. aureus carriage rates, macrolide resistance, disease occurrence, and population structure between samples collected pre- and post-pandemic were examined. By using respiratory samples, chart reviews, and risk factor questionnaires of 498 children from a …

Gambian Mothers Lack Obstetric Danger Sign Knowledge, But Educational Intervention Shows Promise, Kara Shannon, Jocelyn Burridge, Brodus Franklin, Sheena Bhushan, Susan Hilsenbeck, Elena V Petrova, James N'Dow, Ibezimako Iwuh, Sharmila Anandasabapathy, Jeffrey P Wilkinson

Faculty, Staff and Students Publications

BACKGROUND: The Gambia has the 12th highest maternal mortality rate in the world, with 80% of deaths resulting from avoidable causes. Unawareness of pregnancy danger signs (DS) has been shown to be a barrier to seeking obstetric care, while app-based education intervention has shown promise.

OBJECTIVE: We aim to assess patient awareness of DS, identify barriers to awareness, and evaluate potential for implementing smartphone-based technologies for education.

METHODS: A cross-sectional semi-structured survey was administered to Gambian women (n = 100) across five hospitals/health centers. Data and informed consent were collected via an online survey portal. Analysis included bivariate analysis and …

Mfgm-Enriched Whey Displays Antiviral Activity Against Common Pediatric Viruses, Evelien Kramer, Ketki Patil, Vassilis Triantis, Jan A H Bastiaans, Michela Mazzon, Sasirekha Ramani, Tim T Lambers

Faculty, Staff and Students Publications

BACKGROUND: Among the most common mucosal viral infections in infants are rotavirus, one of the main causes of severe gastroenteritis in infants and children up to 5 years, and respiratory syncytial virus (RSV), one of the leading causes of lower respiratory tract infections. Both human milk and bovine milk derived factors may provide protection against mucosal viral infections. More recently, a similar activity of milk derived proteins was suggested for SARS-CoV-2. The goal of the current study was to test antiviral activity of the bovine milkfat globule membrane (MFGM) against rotavirus, RSV and SARS-CoV-2 and to further characterize MFGM-enriched whey …

Autophagy Determines Distinct Cell Fates In Human Amnion And Chorion Cells, Mary Elise L Severino, Lauren Richardson, Ananth Kumar Kammala, Enkhtuya Radnaa, Kamil Khanipov, Leslie Michelle M Dalmacio, Indira U Mysorekar, Marian Kacerovsky, Ramkumar Menon

Faculty, Staff and Students Publications

Human fetal membranes (amniochorion) that line the intrauterine cavity consist of two distinct cell layers; single-layer amnion epithelial cells (AEC) and multilayer chorion trophoblast cells (CTC). These layers are connected through a collagen-rich extracellular matrix. Cellular remodeling helps support membrane growth and integrity during gestation and helps to maintain pregnancy. Preterm prelabor rupture of the human amniochorionic (fetal) membrane (pPROM) is antecedent to 40% of all spontaneous preterm birth. Oxidative stress (OS) induced activation of the p38 MAPK due to various maternal risk exposures and the amniochorion cells' senescence are reported pathological features of pPROM. Our transcriptomics analysis implicated dysregulated …

Select Gut Microbiota Impede Rotavirus Vaccine Efficacy, Vu L Ngo, Yanling Wang, Yadong Wang, Zhenda Shi, Robert Britton, Jun Zou, Sasirekha Ramani, Baoming Jiang, Andrew T Gewirtz

Faculty, Staff and Students Publications

BACKGROUND & AIMS: The protection provided by rotavirus (RV) vaccines is highly heterogeneous among individuals. We hypothesized that microbiota composition might influence RV vaccine efficacy.

METHODS: First, we examined the potential of segmented filamentous bacteria (SFB) colonization to influence RV vaccine efficacy in mice. Next, we probed the influence of human microbiomes on RV vaccination via administering mice fecal microbial transplants (FMTs) from children with robust or minimal RV vaccine responsiveness. Post-FMT, mice were subjected to RV vaccination followed by RV challenge.

RESULTS: SFB colonization induced a phenotype that was reminiscent of RV vaccine failure (ie, failure to generate RV …

Pik-Iii Exerts Anti-Fibrotic Effects In Activated Fibroblasts By Regulating P38 Activation, Santiago Sanchez, Aaron K Mcdowell-Sanchez, Sharaz B Al-Meerani, Juan D Cala-Garcia, Alan R Waich Cohen, Scott A Ochsner, Neil J Mckenna, Lindsay J Celada, Minghua Wu, Shervin Assassi, Ivan O Rosas, Konstantin Tsoyi

Faculty, Staff and Students Publications

Systemic sclerosis (SSc), also known as scleroderma, is an autoimmune-driven connective tissue disorder that results in fibrosis of the skin and internal organs such as the lung. Fibroblasts are known as the main effector cells involved in the progression of SSc through the induction of extracellular matrix (ECM) proteins and myofibroblast differentiation. Here, we demonstrate that 4'-(cyclopropylmethyl)-N2-4-pyridinyl-[4,5'-bipyrimidine]-2,2'-diamine (PIK-III), known as class III phosphatidylinositol 3-kinase (PIK3C3/VPS34) inhibitor, exerts potent antifibrotic effects in human dermal fibroblasts (HDFs) by attenuating transforming growth factor-beta 1 (TGF-β1)-induced ECM expression, cell contraction and myofibroblast differentiation. Unexpectedly, neither genetic silencing of PIK3C3 nor other PIK3C3 inhibitors (e.g., …

Implementation Of A Targeted “Micro-Learning” Inpatient Diabetes Education Program In A Rural Hospital Setting, Melissa Lynn Fannon-Wisner

Graduate Theses, Dissertations, and Problem Reports (ETD)

Introduction/Background: Diabetes affects a large portion of Americans and West Virginians annually. Inpatient hospitalization provides an opportunity to provide evidence-based diabetes education to patients. A lack of access to diabetes education at rural hospitals across the United States exists, creating a need for patients who require education while hospitalized. An intervention needs to be explored to bridge the gap for patients that need education when the diabetes educator is not available.

Purpose: The purpose of this Doctor of Nursing Practice (DNP) project is to create and implement a video micro-learning diabetic education series that can be utilized for diabetes self-management …

Carfilzomib-Loaded Ternary Polypeptide Nanoparticles Stabilized By Polycationic Complexation, Preye Mike Agbana, Ji Eun Park, Piotr G. Rychahou, Kyung-Bo Kim, Younsoo Bae

Markey Cancer Center Faculty Publications

Carfilzomib (CFZ) is a second-generation proteasome inhibitor showing great efficacy in multiple myeloma treatment, yet its clinical applications for other diseases such as solid cancers are limited due to low aqueous solubility and poor biostability. Ternary polypeptide nanoparticles (tPNPs) are drug carriers that we previ- ously reported to overcome these pharmaceutical limitations by entrapping CFZ in the core of the nanopar- ticles and protecting the drugs from degradation in biological media. However, preclinical studies revealed that tPNPs would require further improvement in particle stability to suppress initial burst drug release and thus achieve prolonged inhibition of proteasome activity with CFZ …

A Community-Engaged, Mixed-Methods Approach To Prioritizing Needs In A Statewide Assessment Of Community Cancer Needs, Jessica R. Thompson, Todd Burus, Caree R. Mcafee, Christine Stroebel, Madeline Brown, Keeghan Francis, Melinda Rogers, Jennifer Redmond Knight, Elaine Russell, Connie L. Sorrell, Elizabeth A. Westbrook, Pamela C. Hull

Markey Cancer Center Faculty Publications

Introduction Kentucky has the highest all-site cancer incidence and death rate in the US. In 2021, the University of Kentucky Markey Cancer Center convened a steering committee to conduct a statewide community cancer needs assessment (CNA). The goal of the final CNA phase was to gather community input on prioritizing Kentucky’s cancer-related needs and ways to address them. Methods In 2021, we recruited 162 people to participate in online concept mapping, a participatory mixed method, to explore connections and identify priority areas. Fifty-one community members and 111 organizational partners participated in survey-based activities to prioritize 80 items representing key CNA …

Impact Of Age >70 Years On Oncological Outcomes In Patients With Non-Muscle-Invasive Bladder Cancer Treated With Bacillus Calmette-Guérin., Roberto Contieri, Valentina Grajales, Wei Shen Tan, Alberto Martini, Akshay Sood, Patrick J. Hensley, Kelly Bree, Niyati Lobo, Graciela M. Nogueras-Gonzalez, Charles C. Guo, Neema Navai, Colin P. Dinney, Ashish M. Kamat

Markey Cancer Center Faculty Publications

OBJECTIVE: To evaluate the impact of age on oncological outcomes in a large contemporary cohort of patients with non-muscle-invasive bladder cancer (NMIBC) treated with adequate Bacillus Calmette-Guérin (BCG).

PATIENTS AND METHODS: We performed an Institutional Review Board-approved retrospective study analysing patients with NMIBC treated with adequate BCG at our institution from 2000 to 2020. Adequate BCG was defined as per United States Food and Drug Administration (FDA) guidelines as being receipt of at least five of six induction BCG instillations with a minimum of two additional doses (of planned maintenance or of re-induction) of BCG instillations within a span of …

Retinoic Acid-Mediated Inhibition Of Mouse Coronavirus Replication Is Dependent On Irf3 And Camkk, Justin H. Franco, Ryan A. Harris, William G. Ryan, Roger Travis Taylor, Robert E. Mccullumsmith, Saurabh Chattopadhyay, Zhixing K. Pan

Markey Cancer Center Faculty Publications

The ongoing COVID-19 pandemic has revealed the shortfalls in our understanding of how to treat coronavirus infections. With almost 7 million case fatalities of COVID-19 globally, the catalog of FDA-approved antiviral therapeutics is limited compared to other medications, such as antibiotics. All-trans retinoic acid (RA), or activated vitamin A, has been studied as a potential therapeutic against coronavirus infection because of its antiviral properties. Due to its impact on different signaling pathways, RA’s mechanism of action during coronavirus infection has not been thoroughly described. To determine RA’s mechanism of action, we examined its effect against a mouse coronavirus, mouse hepatitis …

Sedimentation Of Large, Soluble Proteins Up To 140 Kda For 1h-Detected Mas Nmr And 13c Dnp Nmr – Practical Aspects, Dallas Bell, Florian Lindemann, Lisa Gerland, Hanna Aucharova, Alexander Klein, Daniel Friedrich, Matthias Hiller, Kristof Grohe, Tobias Meier, Barth Van Rossum, Anne Diehl, Jon Hughes, Leonard J. Mueller, Rasmus Linser, Anne-Frances Miller, Hartmut Oschkinat

Markey Cancer Center Faculty Publications

Solution NMR is typically applied to biological systems with molecular weights < 40 kDa whereas magic-angle-spinning (MAS) solid-state NMR traditionally targets very large, oligomeric proteins and complexes exceeding 500 kDa in mass, including fibrils and crystalline protein preparations. Here, we propose that the gap between these size regimes can be filled by the approach presented that enables investigation of large, soluble and fully protonated proteins in the range of 40–140 kDa. As a key step, ultracentrifugation produces a highly concentrated, gel-like state, resembling a dense phase in spontaneous liquid-liquid phase separation (LLPS). By means of three examples, a Sulfolobus acidocaldarius bifurcat- ing electron transfer flavoprotein (SaETF), tryptophan synthases from Salmonella typhimurium (StTS) and their dimeric β-subunits from Pyrococcus furiosus (PfTrpB), we show that such samples yield well-resolved proton-detected 2D and 3D NMR spectra at 100 kHz MAS without heterogeneous broadening, similar to diluted liquids. Herein, we provide practical guidance on centrifugation conditions and tools, sample behavior, and line widths expected. We demonstrate that the observed chemical shifts correspond to those obtained from μM/low mM solutions or crystalline samples, indicating structural integrity. Nitrogen line widths as low as 20–30 Hz are observed. The presented approach is advantageous for proteins or nucleic acids that cannot be deuterated due to the expression system used, or where relevant protons cannot be re-incorporated after expression in deuterated medium, and it circumvents crystallization. Importantly, it allows the use of low-glycerol buffers in dynamic nuclear polarization (DNP) NMR of proteins as demonstrated with the cyanobacterial phytochrome Cph1.

Spinal Fusion Enhanced Recovery After Surgery, Children's Mercy Kansas City

Clinical Pathways

No abstract provided.