Digital Commons Network^™

Caspase-8 Scaffolding Function And Mlkl Regulate Nlrp3 Inflammasome Activation Downstream Of Tlr3., Seokwon Kang, Teresa Fernandes-Alnemri, Corey Rogers, Lindsey Mayes, Ying Wang, Christopher Dillon, Linda Roback, William Kaiser, Andrew Oberst, Junji Sagara, Katherine A Fitzgerald, Douglas R Green, Jianke Zhang, Edward S Mocarski, Emad S Alnemri

Department of Biochemistry and Molecular Biology Faculty Papers

TLR2 promotes NLRP3 inflammasome activation via an early MyD88-IRAK1-dependent pathway that provides a priming signal (signal 1) necessary for activation of the inflammasome by a second potassium-depleting signal (signal 2). Here we show that TLR3 binding to dsRNA promotes post-translational inflammasome activation through intermediate and late TRIF/RIPK1/FADD-dependent pathways. Both pathways require the scaffolding but not the catalytic function of caspase-8 or RIPK1. Only the late pathway requires kinase competent RIPK3 and MLKL function. Mechanistically, FADD/caspase-8 scaffolding function provides a post-translational signal 1 in the intermediate pathway, whereas in the late pathway it helps the oligomerization of RIPK3, which together with …

The Sox9 Upstream Region Prone To Chromosomal Aberrations Causing Campomelic Dysplasia Contains Multiple Cartilage Enhancers, Baojin Yao, Qiuqing Wang, Chia-Feng Liu, Pallavi Bhattaram, Wei Li, Timothy J. Mead, James F. Crish, Véronique Lefebvre

Biomedical Sciences

Two decades after the discovery that heterozygous mutations within and around SOX9 cause campomelic dysplasia, a generalized skeleton malformation syndrome, it is well established that SOX9 is a master transcription factor in chondrocytes. In contrast, the mechanisms whereby translocations in the –350/–50-kb region 5 of SOX9 cause severe disease and whereby SOX9 expression is specified in chondrocytes remain scarcely known. We here screen this upstream region and uncover multiple enhancers that activate Sox9-promoter transgenes in the SOX9 expression domain. Three of them are primarily active in chondrocytes. E250 (located at – 250 kb) confines its activity to condensed prechondrocytes, …

Gonad: Genome-Editing Via Oviductal Nucleic Acids Delivery System: A Novel Microinjection Independent Genome Engineering Method In Mice., Gou Takahashi, Channabasavaiah B. Gurumurthy, Kenta Wada, Hiromi Miura, Masahiro Sato, Masato Ohtsuka

Journal Articles: Genetics, Cell Biology & Anatomy

Microinjection is considered the gold standard technique for delivery of nucleic acids (NAs; transgenes or genome editing tools such as CRISPR/Cas9 systems) into embryos, for creating genetically modified organisms. It requires sophisticated equipment as well as well-trained and highly skilled personnel to perform the micro-injection technique. Here, we describe a novel and simple microinjection-independent technique, called Genome-editing via Oviductal Nucleic Acids Delivery (GONAD). Using GONAD, we show that NAs (e.g., eGFP mRNA or Cas9 mRNA/sgRNAs) can be effectively delivered to pre-implantation embryos within the intact mouse oviduct by a simple electroporation method, and result in the desired genetic modification in …

Telemetric Blood Pressure Assessment In Angiotensin Ii-Infused Apoe-/- Mice: 28 Day Natural History And Comparison To Tail-Cuff Measurements, Christopher M. Haggerty, Andrea C. Mattingly, Ming C. Gong, Wen Su, Alan Daugherty, Brandon K. Fornwalt

Saha Cardiovascular Research Center Faculty Publications

Abdominal aortic aneurysm (AAA) is a disease of the aortic wall, which can progress to catastrophic rupture. Assessment of mechanical characteristics of AAA, such as aortic distensibility, may provide important insights to help identify at-risk patients and understand disease progression. While the majority of studies on this topic have focused on retrospective patient data, recent studies have used mouse models of AAA to prospectively evaluate the evolution of aortic mechanics. Quantification of aortic distensibility requires accurate measurement of arterial blood pressure, particularly pulse pressure, which is challenging to perform accurately in murine models. We hypothesized that volume/pressure tail-cuff measurements of …

Antibacterial Characterization Of Novel Synthetic Thiazole Compounds Against Methicillin-Resistant Staphylococcus Pseudintermedius, Haroon Mohammad, P.V. N. Reddy, Dennis Monteleone, Abdelrahman S. Mayhoub, Mark S. Cushman, G. Kenitra Hammac, Mohamed N. Seleem

Department of Comparative Pathobiology Faculty Publications

Staphylococcus pseudintermedius is a commensal organism of companion animals that is a significant source of opportunistic infections in dogs. With the emergence of clinical isolates of S. pseudintermedius (chiefly methicillin-resistant S. pseudintermedius (MRSP)) exhibiting increased resistance to nearly all antibiotic classes, new antimicrobials and therapeutic strategies are urgently needed. Thiazole compounds have been previously shown to possess potent antibacterial activity against multidrug-resistant strains of Staphylococcus aureus of human and animal concern. Given the genetic similarity between S. aureus and S. pseudintermedius, this study explores the potential use of thiazole compounds as novel antibacterial agents against methicillin-sensitive S. pseudintermedius (MSSP) and …

Low Level Lead Exposure Impairs Attentional Set Shifting Task Performance Depending Upon Sex And Developmental Periods Of Exposure, L.S. Neuwirth, D.W. Anderson, J. S. Schneider

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Introduction

Exposure to low levels of lead (Pb) impairs a variety of cognitive processes. Although children exposed to Pb developmentally present with a variety of cognitive impairments that include deficits in learning, memory, language, and executive functioning, experimental work on Pb toxicity in rats has focused mostly on learning and memory deficits and less on executive functions. However, detrimental effects on executive functioning could lead to or even underlie a variety of other cognitive problems attributed to Pb exposure. In this study, we examined the ability of Long Evans rats (control and Pb-exposed: 150ppm Pb-acetate in food given perinatally (gestation …

Amyloid Peptide Inactivating Enzyme To Treat Alzheimer's Disease Peripherally, Louis B. Hersh, Hanjun Guan

Molecular and Cellular Biochemistry Faculty Patents

Methods for treatment and/or prevention of Alzheimer's disease comprising inactivating peripheral AP in serum to a reduce A(3 in the brain. Methods comprise expression of amyloid peptide inactivating enzyme on bone marrow cells; and coupling of amyloid peptide inactivating enzyme to hematopoietic cells.

Fructose Mediated Non-Alcoholic Fatty Liver Is Attenuated By Ho-1-Sirt1 Module In Murine Hepatocytes And Mice Fed A High Fructose Diet, Komal Sodhi, Nitin Puri, Gaia Favero, Sarah Stevens, Charles Meadows, Nader G. Abraham, Rita Rezzani, Hayden A. Ansinelli, Edward Lebovics, Joseph I. Shapiro

Biochemistry and Microbiology

Background

Oxidative stress underlies the etiopathogenesis of nonalcoholic fatty liver disease (NAFLD), obesity and cardiovascular disease (CVD). Heme Oxygenase-1 (HO-1) is a potent endogenous antioxidant gene that plays a key role in decreasing oxidative stress. Sirtuin1 (SIRT1) belongs to the family of NAD-dependent de-acyetylases and is modulated by cellular redox.

Hypothesis

We hypothesize that fructose-induced obesity creates an inflammatory and oxidative environment conducive to the development of NAFLD and metabolic syndrome. The aim of this study is to determine whether HO-1 acts through SIRT1 to form a functional module within hepatocytes to attenuate steatohepatitis, hepatic fibrosis and cardiovascular dysfunction.

Methods …

Long-Term Hypoxia Alters Ovine Fetal Adrenal Enos And Cortisol Biosynthesis, Elizabeth Anne Newby

Loma Linda University Electronic Theses, Dissertations & Projects

Maintaining normal levels of cortisol in response to chronic stress, while retaining the ability to respond to acute stress, is important for ensuring normal fetal growth and development. Long-term hypoxia (LTH) causes adaptations in the fetal hypothalamopituitary- adrenal (HPA) axis that maintain basal cortisol levels but enhance production in response to a secondary stress. Nitric oxide (NO), produced by endothelial nitric oxide synthase (eNOS) in the adrenal cortex, plays a significant role in regulating cortisol production in the LTH fetus. The production of NO is regulated by eNOS activity which can be altered via phosphorylation through key signaling pathways. In …

Crotalus Snake Venom Preconditioning To Prevent Surgical Brain Injury, Cherine Hee-Sun Kim

Loma Linda University Electronic Theses, Dissertations & Projects

Preventive measures are increasingly relevant to medical practice. Preconditioning, a preemptive therapy that administers mildly harmful stimuli to induce endogenous protective mechanisms before major injury, has been shown to minimize injury in many animal models. Given the elective nature of most neurosurgical procedures, the surgical brain injury (SBI) rodent model provides an ideal platform for preconditioning. Our work shows that preconditioning with Crotalus rattlesnake venom, known for its hemorrhagic and inflammatory effects, mitigates some harmful effects of SBI. We have identified two proteins of interest in Crotalus venom: snake venom metalloproteinase (SVMP), an enzyme with hemorrhagic effects, and phospholipase A2 …

Lipid Regulation As A Critical Factor In The Development Of Alzheimer's Disease, Michael A. Castello

Loma Linda University Electronic Theses, Dissertations & Projects

Alzheimer’s disease (AD) is the most common form of dementia in the United States, representing around eighty percent of all cases. For more than two decades, researchers have been led by the amyloid cascade hypothesis, which assumes that accumulation of the amyloid peptide Aβ, derived by proteolytic processing from the amyloid precursor protein (APP), is the key pathogenic trigger in AD. To date, therapies have largely focused on removing Aβ from the brain, an approach that has produced disappointing clinical outcomes. I present an alternative hypothesis in which Aβ production and aggregation is a symptom of a larger, systemic disease …

Acclimatization To High-Altitude, Long-Term Hypoxia Alters Bk Channel Structure And Function, Xiaoxiao Tao

Loma Linda University Electronic Theses, Dissertations & Projects

We examined the major possible mechanisms for the left shift of the BK channel I-V relationship in native basilar artery myocytes from the two LTH groups. These mechanisms included: differential expression of the accessary BK -1 subunit; differential phosphorylation of the BK subunit; and splice variation of the BK subunit. Using molecular cloning, heterologous expression, and patch-clamp electrophysiology techniques, we elucidated a mechanism that, at least in part, contributes to the differences we observed between channels from native normoxic and LTH myocytes.

Molecular Characterization And Inhibition Of Antibodies Elicited Against Galactosyltransferase Knockout Pig Xenografts, John M. Stewart

Loma Linda University Electronic Theses, Dissertations & Projects

On average, 21 people in the United States die every day while waiting for a transplantable organ. Moreover, at the end of 2014, nearly 130,000 additional individuals were still on the transplant waiting list. Xenotransplantation, the transplantation of cells and/or organs from a member of another species, is both a viable supplement for, and bridge to, allotransplantation. Pigs are the favored organ donor because they: 1) breed and grow rapidly, 2) can be genetically modified, and 3) present few ethical concerns. However, due in part to their phylogenetic distance, several barriers must be successfully overcome before clinical xenotransplantation can be …

Tslp-Induced Mechanisms And Potential Therapies For Crlf2 B-Cell Acute Lymphoblastic Leukemia, Olivia L. Francis

Loma Linda University Electronic Theses, Dissertations & Projects

Childhood CRLF2 B-cell Acute Lymphoblastic Leukemia (CRLF2 B-ALL) is a high-risk form of leukemia that is associated with poor patient survival outcomes. CRLF2 B-ALL is five times more prevalent in Hispanic children than others and is associated with a higher rate of relapse, thus contributing significantly to childhood cancer health disparities. This disease occurs due to alterations of the CRLF2 gene, leading to overexpression of the CRLF2 protein- a component of the receptor signaling complex for the cytokine Thymic Stromal LymphoPoietin (TSLP) on the surface of B-ALL cells. TSLP has been shown to induce proliferation of human and mouse B-cell …

Nonparametric Inference In Hidden Markov Models Using P-Splines, Roland Langrock, Thomas Kneib, Alexander Sohn, Stacy L. Deruiter

University Faculty Publications and Creative Works

Hidden Markov models (HMMs) are flexible time series models in which the distribution of the observations depends on unobserved serially correlated states. The state-dependent distributions in HMMs are usually taken from some class of parametrically specified distributions. The choice of this class can be difficult, and an unfortunate choice can have serious consequences for example on state estimates, and more generally on the resulting model complexity and interpretation. We demonstrate these practical issues in a real data application concerned with vertical speeds of a diving beaked whale, where we demonstrate that parametric approaches can easily lead to overly complex state …

Molecular Mechanisms That Govern Human Cardiac Stem Cell Age Disparity, Tania Fuentes

Loma Linda University Electronic Theses, Dissertations & Projects

Transplantation of adult endogenous cardiovascular progenitor cells for heart repair results in some clinical benefit, however these stem cells lack the regenerative capacity unique to neonatal cardiovascular stem cells. The goal of this work was to identify mechanisms that contribute to the decline of cardiac stem cell regenerative ability with age and investigate novel therapeutic strategies to improve cardiac stem cell function. When comparing neonatal and adult cardiovascular stem cell clones, both cell types were capable of cardiomyogenic differentiation. However, the expression levels of forty-one microRNAs were significantly altered with age. Expression differences were correlated with reduced proliferation and a …

Identifying The Molecular Mechanisms Of Early Cachexia Using Whole Transcriptome Sequencing In Muscle And Fat Biopsies From Cancer Patients, Amal Hussain Al Hadad

Dissertations

Cancer cachexia is responsible for one third of cancer–related deaths and contributes to the death of many others. More than 80% of cancer patients are cachectic towards the end of life. Despite intensive research, the mechanisms of cancer cachexia are still poorly understood. It is our hypothesis that identification of early changes in gene expression in cachexia will lead to an improved understanding of the mechanism that trigger this important problem in cancer patients. Thus, to shed light on the mechanisms involved in the major cachexia target tissues, we investigated the entire transcriptome in muscle and fat to identify altered …

Vitamin D As An Adjunctive Therapy In Asthma. Part 1: A Review Of Potential Mechanisms, Conor Kerley, Basil Elnazir, John Faul, Liam Cormican

Articles

Vitamin D deficiency (VDD) is highly prevalent worldwide. The classical role for vitamin D is to regulate calcium absorption form the gastrointestinal tract and influence bone health. Recently vitamin D receptors and vitamin D metabolic enzymes have been discovered in numerous sites systemically supporting diverse extra-skeletal roles of vitamin D, for example in asthmatic disease. Further, VDD and asthma share several common risk factors including high latitude, winter season, industrialization, poor diet, obesity, and dark skin pigmentation. Vitamin D has been demonstrated to possess potent immunomodulatory effects, including effects on T cells and B cells as well as increasing production …

Vitamin D As An Adjunctive Therapy In Asthma. Part 2: A Review Of Human Studies, Conor Kerley, Basil Elnazir, John Faul, Liam Cormican

Articles

Vitamin D deficiency (VDD) is highly prevalent worldwide, with adverse effects on bone health but also potentially other unfavorable consequences. VDD and asthma-incidence/severity share many common risk factors, including winter season, industrialization, poor diet, obesity, dark skin pigmentation, and high latitude. Multiple anatomical areas relevant to asthma contain both the enzyme responsible for producing activated vitamin D and the vitamin D receptor suggesting that activated vitamin D (1,25- dihydroxyvitamin D) may have important local effects at these sites. Emerging evidence suggests that VDD is associated with increased airway hyperresponsiveness, decreased pulmonary function, worse asthma control, and possibly decreased response to …

Pseudomonas Aeruginosa Reduces Vx-809 Stimulated F508del-Cftr Chloride Secretion By Airway Epithelial Cells, Bruce A. Stanton, Bonita Coutermarsh, Roxanna Barnaby, Deborah Hogan

Dartmouth Scholarship

Background:

P. aeruginosa is an opportunistic pathogen that chronically infects the lungs of 85% of adult patients with Cystic Fibrosis (CF). Previously, we demonstrated that P. aeruginosa reduced wt-CFTR Cl secretion by airway epithelial cells. Recently, a new investigational drug VX-809 has been shown to increase F508del-CFTR Cl secretion in human bronchial epithelial (HBE) cells, and, in combination with VX-770, to increase FEV1 (forced expiratory volume in 1 second) by an average of 3-5% in CF patients homozygous for the F508del-CFTR mutation. We propose that P. aeruginosa infection of CF lungs reduces VX-809 + VX-770- stimulated F508del-CFTR Cl secretion, and …

Astrocyte Morphology, Heterogeneity, And Density In The Developing African Giant Rat (Cricetomys Gambianus), Matthew A. Olude, Oluwaseun A. Mustapha, Oluwatunde A. Aderounmu, James O. Olopade, Amadi O. Ihunwo

Brain and Mind Institute

Astrocyte morphologies and heterogeneity were described in male African giant rats (AGR; Cricetomys gambianus, Waterhouse) across three age groups (five neonates, five juveniles, and five adults) using Silver impregnation method and immunohistochemistry against glial fibrillary acidic protein. Immunopositive cell signaling, cell size and population were least in neonates, followed by adults and juveniles, respectively. In neonates, astrocyte processes were mostly detected within the glia limitans of the mid and hind brain; their cell bodies measuring 32 ± 4.8 μm in diameter against 91 ± 5.4 μm and 75 ± 1.9 μm in juveniles and adults, respectively. Astrocyte heterogeneity in …

Intestinal Colonization Dynamics Of Vibrio Cholerae, Salvador Almagro-Moreno, Kali Pruss, Ronald K. Taylor

Dartmouth Scholarship

To cause the diarrheal disease cholera, Vibrio cholerae must effectively colonize the small intestine. In order to do so, the bacterium needs to successfully travel through the stomach and withstand the presence of agents such as bile and antimicrobial peptides in the intestinal lumen and mucus. The bacterial cells penetrate the viscous mucus layer covering the epithelium and attach and proliferate on its surface. In this review, we discuss recent developments and known aspects of the early stages of V. cholerae intestinal colonization and highlight areas that remain to be fully understood. We propose mechanisms and postulate a model …

Angiotensin Ii Is Required To Induce Exaggerated Salt Sensitivity In Dahl Rats Exposed To Maternal Separation, Analia S. Loria, David M. Pollock, Jennifer S. Pollock

Pharmacology and Nutritional Sciences Faculty Publications

We previously reported that maternal separation, rat model of early life stress, enhances pressor responses to acute and chronic stressors. The aims of this study were to determine whether Dahl salt-sensitive (DS) rats subjected to maternal separation (MatSep-DS) as compared to normally reared DS (Ctl-DS) rats show exaggerated blood pressure responses to acute behavioral stressors, such as restraint stress or air jet stress (AJS), or, hypertensive stimuli including chronic high-salt diet (4% NaCl) and angiotensin II (AngII) infusion (200 ng/Kg/min) during 1 week. MatSep was performed in male DS rats for 3 h/day from postnatal days 2-14. At 8 weeks …

Transport Of Bovine Milk Exosomes In Human Endothelial Cell, Rio Jati Kusuma

Department of Nutrition and Health Sciences: Dissertations, Theses, and Student Research

MicroRNAs (miRNAs) can be synthesized endogenously and cause gene repression. Encapsulation of miRNAs in exosomes confers protection against degradation and a vehicle for shuttling between cells and tissues, and cellular uptake by endocytosis. Exosomes can be found in biological fluids and foods including milk. Evidence suggests that humans absorb cow’s milk exosomes and deliver the microRNA cargo to peripheral tissues, consistent with gene regulation by nucleic acids across species boundaries. There is strong evidence that milk exosomes may cross the mucosa without re-packaging in mice. Here, we tested the hypothesis that human vascular endothelial cells transport milk exosomes by endocytosis, …

Antibacterial Evaluation Of Polygonum Multiflorum, Andy S. Demianicz

Seton Hall University Dissertations and Theses (ETDs)

Polygonum multiflorum, commonly known as Chinese Knotweed, is a tonic herb primarily used to enhance bodily functions. Recently, it has been shown to contain strong antioxidant, antibacterial and antiviral activity. The extract from the roots of Chinese Knotweed was used to assess its antibacterial properties against a broad spectrum of bacterial species, including 5 Gram-positive (B. cereus, B. megaterium, S. epidermidis, S. mutans and S. pyogenes) and 4 Gram-negative (E. aerogenes, E. coli, P. vulgaris and P. aeruginosa). Microtiter assays were carried out to evaluate the antibacterial activity of Chinese Knotweed at 0.5%, 1%, 1.5% and 2.5% concentrations in order …

Effect Of Chinese Knotweed Extract On Hsv-1 Infection Of Vero Cells, Derek J. Prince

Seton Hall University Dissertations and Theses (ETDs)

HSV-1 is an opportunistic virus responsible for infecting the majority of the global population. The ability of this virus to strategically alternate between lytic and latent states has made the development of effective treatments challenging. Polygonum multiflorum (Chinese Knotweed, CK) is a perennial plant local to the south central regions of China. The thunb (root) constituents of this compound have been used in traditional Chinese medicines to treat a broad range of illnesses for centuries. In the present study, CK root extracts were evaluated for potential anti-HSV-1 activity. Cytopathic effect (CPE) monitoring studies indicate that Vero cells infected with HSV-1 …

Cerium Oxide Nanoparticles Inhibit Lipopolysaccharide Inducedmap Kinase/Nf-Kb Mediated Severe Sepsis, Mani Maheshwari, Nandini Manne Phd, Shinichi Asano, Eric Blough, Kevin M. Rice, Niraj Nepa,, Erin Fankhanel

Pharmaceutical Science and Research

The life threatening disease of sepsis is associated with high mortality. Septic patient survivability with currently available treatments has failed to improve. The purpose of this study was to evaluate whether lipopolysaccharide (LPS) induced sepsis mortality and associated hepatic dysfunction can be prevented by cerium oxide nanoparticles (CeO2NPs) treatment in male Sprague Dawley rats. Here we provide the information about the methods processing of raw data related to our study published in Biomaterials (Selvaraj et al., Biomaterials, 2015, In press) and Data in Brief (Selvaraj et al., Data in Brief, 2015, In Press). The data present here provides confirmation …

Cannabinoid-Mediated Epigenetic Regulation Of Immune Functions, Jessica Margaret Sido

Theses and Dissertations

The cannabinoid system consisting of exogenous and endogenous ligands as well as dedicated receptors has been proposed to play a regulatory role in immune functions. The exogenous cannabinoid Δ9-tetrahydrocannabinol (THC), one of the most widely studied marijuana derivatives, has been associated with multiple anti-inflammatory properties over the years. The majority of THC research has centered on the shift from Th1 to Th2 responses however, the complexity of inflammation has since increased. Recent studies have revealed that epigenome, Th cell subsets, and immunoregulatory cell induction, are all known to impact inflammation. In the current research, we have attempted to look holistically …

Drug Predictive Cues Activate Aversion-Sensitive Striatal Neurons That Encode Drug Seeking, Daniel S. Wheeler, Mykel A. Robble, Emily M. Hebron, Matthew J. Dupont, Amanda L. Ebben, Robert A. Wheeler

Biomedical Sciences Faculty Research and Publications

Drug-associated cues have profound effects on an addict’s emotional state and drug-seeking behavior. Although this influence must involve the motivational neural system that initiates and encodes the drug-seeking act, surprisingly little is known about the nature of such physiological events and their motivational consequences. Three experiments investigated the effect of a cocaine-predictive stimulus on dopamine signaling, neuronal activity, and reinstatement of cocaine seeking. In all experiments, rats were divided into two groups (paired and unpaired), and trained to self-administer cocaine in the presence of a tone that signaled the immediate availability of the drug. For rats in the paired group, …

Tonb Not Directly Related To Efflux Of Antibiotics In E. Coli, Amber Gombash

Honors Projects

Studies in Pseudomonas aeruginosa have suggested that the TonB energy transduction system directly contributes to efflux-mediated antibiotic resistance, ostensibly by energizing one or more efflux systems. We have found ∆tonB strains of Escherichia coli to similarly be more sensitive to certain antibiotics relative to wild-type strains. To test the hypothesis that this enhanced sensitivity involved the energization of efflux systems, sensitivity patterns for a variety of antibiotics were evaluated using a set of strains differentially lacking genes encoding the Acr efflux system, the universal outer membrane efflux portal TolC, and TonB. No correlation was evident between the resistance phenotypes of …