Digital Commons Network^™

Comprehensive Analysis Of The Secreted Proteome Of Adult Necator Americanus Hookworms, Jayden Logan, Young-Jun Choi, Makedonka Mitreva, Et Al

Open Access Publications

The human hookworm Necator americanus infects more than 400 million people worldwide, contributing substantially to the poverty in these regions. Adult stage N. americanus live in the small intestine of the human host where they inject excretory/secretory (ES) products into the mucosa. ES products have been characterized at the proteome level for a number of animal hookworm species, but until now, the difficulty in obtaining sufficient live N. americanus has been an obstacle in characterizing the secretome of this important human pathogen. Herein we describe the ES proteome of N. americanus and utilize this information along with RNA Seq data …

The Trajectory Of Gait Development In Mice, Shyam K Akula, Katherine B Mccullough, Claire Weichselbaum, Joseph D Dougherty, Susan E Maloney

Open Access Publications

OBJECTIVE: Gait irregularities are prevalent in neurodevelopmental disorders (NDDs). However, there is a paucity of information on gait phenotypes in NDD experimental models. This is in part due to the lack of understanding of the normal developmental trajectory of gait maturation in the mouse.

MATERIALS AND METHODS: Using the DigiGait system, we have developed a quantitative, standardized, and reproducible assay of developmental gait metrics in commonly used mouse strains that can be added to the battery of mouse model phenotyping. With this assay, we characterized the trajectory of gait in the developing C57BL/6J and FVB/AntJ mouse lines.

RESULTS: In both …

Nodal And Planar Cell Polarity Signaling Cooperate To Regulate Zebrafish Convergence And Extension Gastrulation Movements, Margot L.K. Williams, Lilianna Solnica-Krezel

Open Access Publications

During vertebrate gastrulation, convergence and extension (C and E) of the primary anteroposterior (AP) embryonic axis is driven by polarized mediolateral (ML) cell intercalations and is influenced by AP axial patterning. Nodal signaling is essential for patterning of the AP axis while planar cell polarity (PCP) signaling polarizes cells with respect to this axis, but how these two signaling systems interact during C and E is unclear. We find that the neuroectoderm of Nodal-deficient zebrafish gastrulae exhibits reduced C and E cell behaviors, which require Nodal signaling in both cell- and non-autonomous fashions. PCP signaling is partially active in Nodal-deficient …

Cd4+ T Cell Expression Of The Il-10 Receptor Is Necessary For Facial Motoneuron Survival After Axotomy, Elizabeth M Runge, Abhirami K Iyer, Deborah O Setter, Felicia M Kennedy, Virginia M Sanders, Kathryn J Jones

Open Access Publications

BACKGROUND: After peripheral nerve transection, facial motoneuron (FMN) survival depends on an intact CD4+ T cell population and a central source of interleukin-10 (IL-10). However, it has not been determined previously whether CD4+ T cells participate in the central neuroprotective IL-10 cascade after facial nerve axotomy (FNA).

METHODS: Immunohistochemical labeling of CD4+ T cells, pontine vasculature, and central microglia was used to determine whether CD4+ T cells cross the blood-brain barrier and enter the facial motor nucleus (FMNuc) after FNA. The importance of IL-10 signaling in CD4+ T cells was assessed by performing adoptive transfer of IL-10 receptor beta (IL-10RB)-deficient …

Reinforcement Biases Subsequent Perceptual Decisions When Confidence Is Low, A Widespread Behavioral Phenomenon, Armin Lak, Emily Hueske, Junya Hirokawa, Paul Masset, Torben Ott, Anne E. Urai, Tobias H. Donner, Matteo Carandini, Susumu Tonegawa, Naoshige Uchida, Adam Kepecs

Open Access Publications

Learning from successes and failures often improves the quality of subsequent decisions. Past outcomes, however, should not influence purely perceptual decisions after task acquisition is complete since these are designed so that only sensory evidence determines the correct choice. Yet, numerous studies report that outcomes can bias perceptual decisions, causing spurious changes in choice behavior without improving accuracy. Here we show that the effects of reward on perceptual decisions are principled: past rewards bias future choices specifically when previous choice was difficult and hence decision confidence was low. We identified this phenomenon in six datasets from four laboratories, across mice, …

Cd300lf Is The Primary Physiologic Receptor Of Murine Norovirus But Not Human Norovirus, Vincent R. Graziano, Forrest C. Walker, Elizabeth A. Kennedy, Ebrahim Hassan, Arthur S. Kim, Megan T. Baldridge, Et Al.

Open Access Publications

Murine norovirus (MNoV) is an important model of human norovirus (HNoV) and mucosal virus infection more broadly. Viral receptor utilization is a major determinant of cell tropism, host range, and pathogenesis. The bona fide receptor for HNoV is unknown. Recently, we identified CD300lf as a proteinaceous receptor for MNoV. Interestingly, its paralogue CD300ld was also sufficient for MNoV infection in vitro. Here we explored whether CD300lf is the sole physiologic receptor in vivo and whether HNoV can use a CD300 ortholog as an entry receptor. We report that both CD300ld and CD300lf are sufficient for infection by diverse MNoV strains …

Pharmacologic Inhibition Of Rgd-Binding Integrins Ameliorates Fibrosis And Improves Function Following Kidney Injury, Jeannine Basta, Lynn Robbins, Lisa Stout, Michael J Prinsen, David W Griggs, Michael Rauchman

Open Access Publications

Fibrosis is a final common pathway for many causes of progressive chronic kidney disease (CKD). Arginine-glycine-aspartic acid (RGD)-binding integrins are important mediators of the pro-fibrotic response by activating latent TGF-β at sites of injury and by providing myofibroblasts information about the composition and stiffness of the extracellular matrix. Therefore, blockade of RGD-binding integrins may have therapeutic potential for CKD. To test this idea, we used small-molecule peptidomimetics that potently inhibit a subset of RGD-binding integrins in a murine model of kidney fibrosis. Acute kidney injury leading to fibrosis was induced by administration of aristolochic acid. Continuous subcutaneous administration of CWHM-12, …

Reduced Neurosteroid Potentiation Of Gabaa Receptors In Epilepsy And Depolarized Hippocampal Neurons, Suchitra Joshi, William H Roden, Jaideep Kapur, Laura A Jansen

Open Access Publications

OBJECTIVE: Neurosteroids regulate neuronal excitability by potentiating γ-aminobutyric acid type-A receptors (GABARs). In animal models of temporal lobe epilepsy, the neurosteroid sensitivity of GABARs is diminished and GABAR subunit composition is altered. We tested whether similar changes occur in patients with epilepsy and if depolarization-induced increases in neuronal activity can replicate this effect.

METHODS: We determined GABAR α4 subunit expression in cortical tissue resected from pediatric epilepsy patients. Modulation of human GABARs by allopregnanolone and Ro15-4513 was measured in Xenopus oocytes using whole-cell patch clamp. To extend the findings obtained using tissue from epilepsy patients, we evaluated GABAR expression and …

Differences In The Chitinolytic Activity Of Mammalian Chitinases On Soluble And Insoluble Substrates, Benjamin A Barad, Lin Liu, Roberto E Diaz, Ralp Basilio, Steven J Van Dyken, Richard M Locksley, James S Fraser

Open Access Publications

Chitin is an abundant polysaccharide used by many organisms for structural rigidity and water repulsion. As such, the insoluble crystalline structure of chitin poses significant challenges for enzymatic degradation. Acidic mammalian chitinase, a processive glycosyl hydrolase, is the primary enzyme involved in the degradation of environmental chitin in mammalian lungs. Mutations to acidic mammalian chitinase have been associated with asthma, and genetic deletion in mice increases morbidity and mortality with age. We initially set out to reverse this phenotype by engineering hyperactive acidic mammalian chitinase variants. Using a screening approach with commercial fluorogenic substrates, we identified mutations with consistent increases …

Incomplete Vesicular Docking Limits Synaptic Strength Under High Release Probability Conditions, Gerardo Malagon, Takafumi Miki, Van Tran, Laura C. Gomez, Alain Marty

Open Access Publications

Central mammalian synapses release synaptic vesicles in dedicated structures called docking/release sites. It has been assumed that when voltage-dependent calcium entry is sufficiently large, synaptic output attains a maximum value of one synaptic vesicle per action potential and per site. Here we use deconvolution to count synaptic vesicle output at single sites (mean site number per synapse: 3.6). When increasing calcium entry with tetraethylammonium in 1.5 mM external calcium concentration, we find that synaptic output saturates at 0.22 vesicle per site, not at 1 vesicle per site. Fitting the results with current models of calcium-dependent exocytosis indicates that the 0.22 …

The Sterlet Sturgeon Genome Sequence And The Mechanisms Of Segmental Rediploidization, Kang Du, Chad Tomlinson, Et Al

Open Access Publications

Sturgeons seem to be frozen in time. The archaic characteristics of this ancient fish lineage place it in a key phylogenetic position at the base of the ~30,000 modern teleost fish species. Moreover, sturgeons are notoriously polyploid, providing unique opportunities to investigate the evolution of polyploid genomes. We assembled a high-quality chromosome-level reference genome for the sterlet, Acipenser ruthenus. Our analysis revealed a very low protein evolution rate that is at least as slow as in other deep branches of the vertebrate tree, such as that of the coelacanth. We uncovered a whole-genome duplication that occurred in the Jurassic, early …

Unc-4 Acts To Promote Neuronal Identity And Development Of The Take-Off Circuit In The Drosophila Cns, Haluk Lacin, W. Ryan Williamson, Gwyneth M. Card, James B. Skeath, James W. Truman

Open Access Publications

The

Loss Of H3k36 Methyltransferase Setd2 Impairs V(D)J Recombination During Lymphoid Development, S Haihua Chu, Putzer J Hung, Et Al

Open Access Publications

Repair of DNA double-stranded breaks (DSBs) during lymphocyte development is essential for V(D)J recombination and forms the basis of immunoglobulin variable region diversity. Understanding of this process in lymphogenesis has historically been centered on the study of RAG1/2 recombinases and a set of classical non-homologous end-joining factors. Much less has been reported regarding the role of chromatin modifications on this process. Here, we show a role for the non-redundant histone H3 lysine methyltransferase, Setd2, and its modification of lysine-36 trimethylation (H3K36me3), in the processing and joining of DNA ends during V(D)J recombination. Loss leads to mis-repair of Rag-induced DNA DSBs, …

Long Non-Coding Rna Graslnd Enhances Chondrogenesis Via Suppression Of The Interferon Type Ii Signaling Pathway, Nguyen P.T. Huynh, Catherine C. Gloss, Jeremiah Lorentz, Ruhang Tang, Jonathan M. Brunger, Audrey Mcalinden, Bo Zhang, Farshid Guilak

Open Access Publications

The roles of long noncoding RNAs (lncRNAs) in musculoskeletal development, disease, and regeneration remain poorly understood. Here, we identified the novel lncRNA

Abnormally Increased Carotid Intima Media-Thickness And Elasticity In Patients With Morquio A Disease, Raymond Y Wang, Kyle D Rudser, Donald R Dengel, Nicholas Evanoff, Julia Steinberger, Nina Movsesyan, Robert Garrett, Katherine Christensen, Deborah Boylan, Stephen R Braddock, Marwan Shinawi, Qi Gan, Adriana M Montaño

Open Access Publications

BACKGROUND: Cardiovascular disease frequently causes morbidity and mortality in mucopolysaccharidoses (MPS); however, cardiovascular anatomy and dysfunction in MPS IVA (Morquio A disease) is not well described. Consequently, the study aimed to compare carotid artery structure and elasticity of MPS IVA patients with other MPS patients and healthy control subjects, and quantitate frequency of MPS IVA cardiac structural and functional abnormalities.

METHODS: Prospective, multi-center echocardiogram and carotid ultrasound evaluations of 12 Morquio A patients were compared with other MPS and healthy control subjects. Average differences between groups were adjusted for age, sex, and height with robust variance estimation for confidence intervals …

T Cell Response Kinetics Determines Neuroinfection Outcomes During Murine Hsv Infection, Aisha G Lee, Jason M Scott, Maria Rita Fabbrizi, Xiaoping Jiang, Dorothy K Sojka, Mark J Miller, Megan T Baldridge, Wayne M Yokoyama, Haina Shin

Open Access Publications

Herpes simplex virus-2 (HSV-2) and HSV-1 both can cause genital herpes, a chronic infection that establishes a latent reservoir in the nervous system. Clinically, the recurrence frequency of HSV-1 genital herpes is considerably less than HSV-2 genital herpes, which correlates with reduced neuronal infection. The factors dictating the disparate outcomes of HSV-1 and HSV-2 genital herpes are unclear. In this study, we show that vaginal infection of mice with HSV-1 leads to the rapid appearance of mature DCs in the draining lymph node, which is dependent on an early burst of NK cell-mediated IFN-γ production in the vagina that occurs …

Cooperative Synaptic And Intrinsic Plasticity In A Disynaptic Limbic Circuit Drive Stress-Induced Anhedonia And Passive Coping In Mice, Marco Pignatelli, Hugo A Tejeda, Et Al

Open Access Publications

Stress promotes negative affective states, which include anhedonia and passive coping. While these features are in part mediated by neuroadaptations in brain reward circuitry, a comprehensive framework of how stress-induced negative affect may be encoded within key nodes of this circuit is lacking. Here, we show in a mouse model for stress-induced anhedonia and passive coping that these phenomena are associated with increased synaptic strength of ventral hippocampus (VH) excitatory synapses onto D1 medium spiny neurons (D1-MSNs) in the nucleus accumbens medial shell (NAcmSh), and with lateral hypothalamus (LH)-projecting D1-MSN hyperexcitability mediated by decreased inwardly rectifying potassium channel (IRK) function. …

Genome-Wide Association Study In Two Cohorts From A Multi-Generational Mouse Advanced Intercross Line Highlights The Difficulty Of Replication Due To Study-Specific Heterogeneity, Xinzhu Zhou, Celine L St Pierre, Natalia M Gonzales, Jennifer Zou, Riyan Cheng, Apurva S Chitre, Greta Sokoloff, Abraham A Palmer

Open Access Publications

There has been extensive discussion of the "Replication Crisis" in many fields, including genome-wide association studies (

Defining Stage-Specific Activity Of Potent New Inhibitors Of Cryptosporidium Parvum Growth In Vitro, Lisa J Funkhouser-Jones, Soumya Ravindran, L. David Sibley

Open Access Publications

No abstract provided.

Acetate Coordinates Neutrophil And Ilc3 Responses Against C. Difficile Through Ffar2, José Luís Fachi, Cristiane Sécca, Blanda Di Luccia, Marco Colonna, Et Al

Open Access Publications

Antibiotic-induced dysbiosis is a key predisposing factor for Clostridium difficile infections (CDIs), which cause intestinal disease ranging from mild diarrhea to pseudomembranous colitis. Here, we examined the impact of a microbiota-derived metabolite, short-chain fatty acid acetate, on an acute mouse model of CDI. We found that administration of acetate is remarkably beneficial in ameliorating disease. Mechanistically, we show that acetate enhances innate immune responses by acting on both neutrophils and ILC3s through its cognate receptor free fatty acid receptor 2 (FFAR2). In neutrophils, acetate-FFAR2 signaling accelerates their recruitment to the inflammatory sites, facilitates inflammasome activation, and promotes the release of …

Platelet Activation In Experimental Murine Neonatal Pulmonary Hypertension, Pavel Davizon-Castillo, Ayed Allawzi, Matthew Sorrells, Susan Fisher, Kristina Baltrunaite, Keith Neeves, Eva Nozik-Grayck, Jorge Dipaola, Cassidy Delaney

Open Access Publications

Serotonin (5-HT) contributes to the pathogenesis of experimental neonatal pulmonary hypertension (PH) associated with bronchopulmonary dysplasia (BPD). Platelets are the primary source of circulating 5-HT and is released upon platelet activation. Platelet transfusions are associated with neonatal mortality and increased rates of BPD. As BPD is often complicated by PH, we tested the hypothesis that circulating platelets are activated and also increased in the lungs of neonatal mice with bleomycin-induced PH associated with BPD. Newborn wild-type mice received intraperitoneal bleomycin (3 units/kg) three times weekly for 3 weeks. Platelets from mice with experimental PH exhibited increased adhesion to collagen under …

Tgfβ1 Orchestrates Renal Fibrosis Following Escherichia Coli Pyelonephritis, Teri N Hreha, Christina A Collins, Allyssa L Daugherty, Joy Twentyman, Nitin Paluri, David A Hunstad

Open Access Publications

Renal scarring after pyelonephritis is linked to long-term health risks for hypertension and chronic kidney disease. Androgen exposure increases susceptibility to, and severity of, uropathogenic Escherichia coli (UPEC) pyelonephritis and resultant scarring in both male and female mice, while anti-androgen therapy is protective against severe urinary tract infection (UTI) in these models. This work employed androgenized female C57BL/6 mice to elucidate the molecular mechanisms of post-infectious renal fibrosis and to determine how these pathways are altered by the presence of androgens. We found that elevated circulating testosterone levels primed the kidney for fibrosis by increasing local production of TGFβ1 before …

Age And Tightness Of Repair Are Predictors Of Heel-Rise Height After Achilles Tendon Rupture, Michael R Carmont, Jennifer A Zellers, Annelie Brorsson, Katarina Nilsson-Helander, Jón Karlsson, Karin Grävare Silbernagel

Open Access Publications

Background: Achilles tendon rupture leads to weakness of ankle plantarflexion. Treatment of Achilles tendon rupture should aim to restore function while minimizing weakness and complications of management.

Purpose: To determine the influence of factors (age, sex, body mass index [BMI], weight, time from injury to operative repair, and tightness of repair) in the initial surgical management of patients after an acute Achilles tendon rupture on 12-month functional outcome assessment after percutaneous and minimally invasive repair.

Study Design: Cohort study; Level of evidence, 3.

Methods: From May 2012 to January 2018, patients sustaining an Achilles tendon rupture receiving operative repair were …

The Safety Of Combined Triple Drug Therapy With Ivermectin, Diethylcarbamazine And Albendazole In The Neglected Tropical Diseases Co-Endemic Setting Of Fiji: A Cluster Randomised Trial, Myra Hardy, Josaia Samuela, Mike Kama, Meciusela Tuicakau, Lucia Romani, Margot J. Whitfeld, Christopher L. King, Gary J. Weil, Anneke C. Grobler, Leanne J. Robinson, John M. Kaldor, Andrew C. Steer

Open Access Publications

Lymphatic filariasis has remained endemic in Fiji despite repeated mass drug administration using the well-established and safe combination of diethylcarbamazine and albendazole (DA) since 2002. In certain settings the addition of ivermectin to this combination (IDA) remains a safe strategy and is more efficacious. However, the safety has yet to be described in scabies and soil-transmitted helminth endemic settings like Fiji. Villages of Rotuma and Gau islands were randomised to either DA or IDA. Residents received weight-based treatment unblinded with standard exclusions. Participants were actively found and asked by a nurse about their health daily for the first two days …

How To Stop The Bleed: First Care Provider Model For Developing Public Trauma Response Beyond Basic Hemorrhage Control, Joshua P Bobko, Dylan J Badin, Leila Danishgar, Kate Bayhan, Kevin J Thompson, William J Harris, R. Todd Baldridge, Gerald R Fortuna

Open Access Publications

INTRODUCTION: Since 2013, the First Care Provider (FCP) model has successfully educated the non-medical population on how to recognize life-threatening injuries and perform interventions recommended by the Committee for Tactical Emergency Casualty Care (C-TECC) and the Hartford Consensus in the disaster setting. Recent programs, such as the federal "Stop The Bleed" campaign, have placed the emphasis of public training on hemorrhage control. However, recent attacks demonstrate that access to wounded, recognition of injury, and rapid evacuation are equally as important as hemorrhage control in minimizing mortality. To date, no training programs have produced a validated study with regard to training …

Neural Mechanisms Of Economic Choices In Mice, Masaru Kuwabara, Ningdong Kang, Timothy E. Holy, Camillo Padoa-Schioppa

Open Access Publications

Economic choices entail computing and comparing subjective values. Evidence from primates indicates that this behavior relies on the orbitofrontal cortex. Conversely, previous work in rodents provided conflicting results. Here we present a mouse model of economic choice behavior, and we show that the lateral orbital (LO) area is intimately related to the decision process. In the experiments, mice chose between different juices offered in variable amounts. Choice patterns closely resembled those measured in primates. Optogenetic inactivation of LO dramatically disrupted choices by inducing erratic changes of relative value and by increasing choice variability. Neuronal recordings revealed that different groups of …

Agonist-Selective Recruitment Of Engineered Protein Probes And Of Grk2 By Opioid Receptors In Living Cells, Miriam Stoeber, Damien Jullié, Joy Li, Soumen Chakraborty, Susruta Majumdar, Nevin A. Lambert, Aashish Manglik, Mark Von Zastrow

Open Access Publications

G protein-coupled receptors (GPCRs) signal through allostery, and it is increasingly clear that chemically distinct agonists can produce different receptor-based effects. It has been proposed that agonists selectively promote receptors to recruit one cellular interacting partner over another, introducing allosteric 'bias' into the signaling system. However, the underlying hypothesis - that different agonists drive GPCRs to engage different cytoplasmic proteins in living cells - remains untested due to the complexity of readouts through which receptor-proximal interactions are typically inferred. We describe a cell-based assay to overcome this challenge, based on GPCR-interacting biosensors that are disconnected from endogenous transduction mechanisms. Focusing …

Proteogenomic Characterization Of Endometrial Carcinoma, Yongchao Dou, Daniel Cui Zhou, Alla Karpova, Yige Wu, Song Cao, Matthew A Wyczalkowski, Qingsong Gao, Sunantha Sethuraman, David Mutch, Katherine Fuh, Li Ding, Et Al

Open Access Publications

We undertook a comprehensive proteogenomic characterization of 95 prospectively collected endometrial carcinomas, comprising 83 endometrioid and 12 serous tumors. This analysis revealed possible new consequences of perturbations to the p53 and Wnt/β-catenin pathways, identified a potential role for circRNAs in the epithelial-mesenchymal transition, and provided new information about proteomic markers of clinical and genomic tumor subgroups, including relationships to known druggable pathways. An extensive genome-wide acetylation survey yielded insights into regulatory mechanisms linking Wnt signaling and histone acetylation. We also characterized aspects of the tumor immune landscape, including immunogenic alterations, neoantigens, common cancer/testis antigens, and the immune microenvironment, all of …

An Inducible Cre Mouse Line To Sparsely Target Nervous System Cells, Including Remak Schwann Cells, Darshan Sapkota, Joseph D Dougherty

Open Access Publications

Nerves of the peripheral nervous system contain two classes of Schwann cells: myelinating Schwann cells that ensheath large caliber axons and generate the myelin sheath, and Remak Schwann cells that surround smaller axons and do not myelinate. While tools exist for genetic targeting of Schwann cell precursors and myelinating Schwann cells, such reagents have been challenging to generate specifically for the Remak population, in part because many of the genes that mark this population in maturity are also robustly expressed in Schwann cell precursors. To circumvent this challenge, we utilized BAC transgenesis to generate a mouse line expressing a tamoxifen-inducible …

Continuous Representations Of Speed By Striatal Medium Spiny Neurons, Wambura C. Fobbs, Sebastiano Bariselli, Julia A. Licholai, Nanami L. Miyazaki, Bridget A. Matikainen-Ankney, Meaghan C. Creed, Alexxai V. Kravitz

Open Access Publications

The striatum is critical for controlling motor output. However, it remains unclear how striatal output neurons encode and facilitate movement. A prominent theory suggests that striatal units encode movements in bursts of activity near specific events, such as the start or end of actions. These bursts are theorized to gate or permit specific motor actions, thereby encoding and facilitating complex sequences of actions. An alternative theory has suggested that striatal neurons encode continuous changes in sensory or motor information with graded changes in firing rate. Supporting this theory, many striatal neurons exhibit such graded changes without bursting near specific actions. …