Medicine and Health Sciences Commons^™

Lc-Ms/Ms Identification Of A Bromelain Peptide Biomarker From Ananas Comosus Merr, Eric R. Secor, Steven M. Szczepanek, Linda Guernsey, Prabitha Natarajan, Karim Rezaul, David K. Han, Roger S. Thrall, Lawrence K. Silbart

UCHC Articles - Research

Bromelain (Br) is a cysteine peptidase (GenBank AEH26024.1) from pineapple, with over 40 years of clinical use. The constituents mediating its anti-inflammatory activity are not thoroughly characterized and no peptide biomarker exists. Our objective is to characterize Br raw material and identify peptides in the plasma of Br treated mice. After SDS-PAGE in-gel digestion, Br (VN#3507; Middletown, CT, USA) peptides were analyzed via LC/MS/MS using 95% protein probability, 95% peptide probability, and a minimum peptide number = 5. Br spiked mouse plasma (1 ug/ul) and plasma from i.p. treated mice (12 mg/kg) were assessed using SRM. In Br raw material, …

Bone-Specific Overexpression Of Npy Modulates Osteogenesis, I Matic, B G. Matthews, T Kizivat, J C. Igwe, D J. Adams, Ivo Kalajzic

UCHC Articles - Research

Objectives

Neuropeptide Y (NPY) is a peptide involved in the regulation of appetite and energy homeostasis. Genetic data indicates that NPY decreases bone formation via central and peripheral activities. NPY is produced by various cell types including osteocytes and osteoblasts and there is evidence suggesting that peripheral NPY is important for regulation of bone formation. We sought to investigate the role of bone-derived NPY in bone metabolism.

Methods

We generated a mouse where NPY was over-expressed specifically in mature osteoblasts and osteocytes (Col2.3NPY) and characterized the bone phenotype of these mice in vivo and in vitro.

Results

Trabecular and cortical …

Distinct Mechanisms Mediate Naïve And Memory Cd8 T-Cell Tolerance, Evan R. Jellison, Elizabeth G. Lingenheld, Li Zu, Lynn Puddington, Leo Lefrancois

UCHC Articles - Research

Peripheral tolerance to developmentally regulated antigens is necessary to sustain tissue homeostasis. We have now devised an inducible and reversible system that allows interrogation of T-cell tolerance induction in endogenous naïve and memory CD8 T cells. Our data show that peripheral CD8 T-cell tolerance can be preserved through two distinct mechanisms, antigen addiction leading to anergy for naïve T cells and ignorance for memory T cells. Induction of antigen in dendritic cells resulted in substantial expansion and maintenance of endogenous antigen-specific CD8 T cells. The self-reactive cells initially exhibited effector activity but eventually became unresponsive. Upon antigen removal, the antigen-specific …

Logic Modeling And The Ridiculome Under The Rug, Michael L. Blinov, Ion I. Moraru

UCHC Articles - Research

Logic-derived modeling has been used to map biological networks and to study arbitrary functional interactions, and fine-grained kinetic modeling can accurately predict the detailed behavior of well-characterized molecular systems; at present, however, neither approach comes close to unraveling the full complexity of a cell. The current data revolution offers significant promises and challenges to both approaches - and could bring them together as it has spurred the development of new methods and tools that may help to bridge the many gaps between data, models, and mechanistic understanding.

Have you used logic modeling in your research? It would not be surprising …

Defining The Developmental Parameters Of Temper Loss In Early Childhood: Implications For Developmental Psychopathology, Heide Hullsiek, Margaret J. Briggs-Gowan

UCHC Articles - Research

Abstract

Background

Temper modulation problems are both a hallmark of early childhood and a common mental health concern. Thus, characterizing specific behavioral manifestations of temper loss along a dimension from normative misbehaviors to clinically significant problems is an important step toward identifying clinical thresholds.

Methods

Parent-reported patterns of temper loss were delineated in a diverse community sample of preschoolers (n = 1,490). A developmentally sensitive questionnaire, the Multidimensional Assessment of Preschool Disruptive Behavior (MAP-DB), was used to assess temper loss in terms of tantrum features and anger regulation. Specific aims were: (a) document the normative distribution of temper loss in …

Swimming Dynamics Of The Lyme Disease Spirochete, Dhruv K. Vig, Charles W. Wolgemuth

UCHC Articles - Research

The Lyme disease spirochete, Borrelia burgdorferi, swims by undulating its cell body in the form of a traveling flat-wave, a process driven by rotating internal flagella. We study B. burgdorferi ’s swimming by treating the cell body and flagella as linearly elastic filaments. The dynamics of the cell are then determined from the balance between elastic and resistive forces and moments. We find that planar, traveling waves only exist when the flagella are effectively anchored at both ends of the bacterium and that these traveling flat-waves rotate as they undulate. The model predicts how the undulation frequency is related …

Simultaneous Measurement And Modulation Of Multiple Physiological Parameters In The Isolated Heart Using Optical Techniques, Leslie M. Loew, Ping Yan

UCHC Articles - Research

Whole-heart multi-parametric optical mapping has provided valuable insight into the interplay of electro-physiological parameters, and this technology will continue to thrive as dyes are improved and technical solutions for imaging become simpler and cheaper. Here, we show the advantage of using improved 2nd-generation voltage dyes, provide a simple solution to panoramic multi-parametric mapping, and illustrate the application of flash photolysis of caged compounds for studies in the whole heart. For proof of principle, we used the isolated rat whole-heart model. After characterising the blue and green isosbestic points of di-4-ANBDQBS and di-4-ANBDQPQ, respectively, two voltage and calcium mapping systems are …

Tlr8: The Forgotten Relative Revindicated, Jorge L. Cervantes, Bennett Weinerman, Chaitali Basole, Juan C. Salazar

UCHC Articles - Research

The endosomal Toll-like receptors (TLRs) TLR3, TLR7, TLR8 and TLR9 are important in sensing foreign nucleic acids encountered by phagocytes. Because TLR8 was initially thought to be non-functional in mice, less is known about TLR8 than the genetically and functionally related TLR7. Originally associated with the recognition of single-stranded RNA of viral origin, there is now evidence that human TLR8 is also able to sense bacterial RNA released within phagosomal vacuoles, inducing the production of both nuclear factor (NF)-κB-dependent cytokines and type I interferons (IFNs), such as IFN-β. The functions of TLR8 extend beyond the recognition of foreign pathogens and …

Looking In The Mouth For Noninvasive Gene Expression-Based Methods To Detect Oral, Oropharyngeal, And Systemic Cancer, Alexander J. Adami

UCHC Articles - Research

Noninvasive diagnosis, whether by sampling body fluids, body scans, or other technique, has the potential to simplify early cancer detection. A classic example is Pap smear screening, which has helped to reduce cervical cancer 75% over the last 50 years. No test is error-free; the real concern is sufficient accuracy combined with ease of use. This paper will discuss methods that measure gene expression or epigenetic markers in oral cells or saliva to diagnose oral and pharyngeal cancers, without requiring surgical biopsy. Evidence for lung and other distal cancer detection is also reviewed.

Herpes Simplex Viruses: Mechanisms Of Dna Replication, Sandra K. Weller

UCHC Articles - Research

Herpes simplex virus (HSV) encodes seven proteins necessary for viral DNA synthesis—UL9 (origin-binding protein), ICP8 (single-strand DNA [ssDNA]-binding protein), UL30/UL42 (polymerase), and UL5/UL8/UL52 (helicase/primase). It is our intention to provide an up-to-date analysis of our understanding of the structures of these replication proteins and how they function during HSV replication. The potential roles of host repair and recombination proteins will also be discussed.

Nf-Κb Contributes To The Detrimental Effects Of Social Isolation After Experimental Stroke, Venugopal Reddy Venna, Gillian Weston, Sharon E. Benashski, Sami Tarabishy, Fudong Liu, Jun Li, Lisa H. Conti, Louise D. Mccullough

UCHC Articles - Research

Social isolation (SI) is increasingly recognized as a risk factor for stroke. Individuals with lack of social support systems have an increased incidence of stroke, poorer recovery, and greater functional decline after injury compared to individuals with social support. Attesting to the importance of social factors in stroke outcome is that these same effects can be reproducibly demonstrated in animals; social interaction improves behavioral deficits and reduces damage after experimental stroke, whereas SI enhances injury. The mechanism by which SI exacerbates injury is unclear. We investigated the role of nuclear factor-kappaB (NF-κB) signaling in male mice that were pair housed …

Gabra2 Genotype, Impulsivity, And Body Mass, Lance O. Bauer

UCHC Articles - Research

Background

The goal of this study was to test a hypothesis associating impulsivity with an elevated body mass index (BMI).

Methods

To this end, we examined associations of BMI with putative genetic, neurophysiological, psychiatric, and psychological indicators of impulsivity in 78 women and 74 men formerly dependent on alcohol or drugs. A second analysis was designed to test the replicability of the genetic findings in an independent sample of 109 women and 111 men with a similar history of substance dependence.

Results

The results of the first analysis showed that BMI was positively correlated with Total and Nonplanning Scale Scores …

Active Induction Of Experimental Autoimmune Encephalomyelitis By Mog35-55 Peptide Immunization Is Associated With Differential Responses In Separate Compartments Of The Choroid Plexus, Nivetha Murugesan, Debayon Paul, Yen Lemire, Bandana Shrestha, Shujun Ge, Joel S. Pachter

UCHC Articles - Research

Background

There is increasing awareness that, aside from producing cerebrospinal fluid, the choroid plexus (CP) might be a key regulator of immune activity in the central nervous system (CNS) during neuroinflammation. Specifically, the CP has recently been posited to control entry of sentinel T cells into the uninflamed CNS during the early stages of neuroinflammatory diseases, like multiple sclerosis (MS) and its animal model experimental autoimmune encephalomyelitis (EAE). As the CP is compartmentalized into a stromal core containing fenestrated capillaries devoid of typical blood–brain barrier properties, surrounded by a tight junction-expressing choroidal epithelium, each of these compartments might mount unique …

Fast Rebinding Increases Dwell Time Of Src Homology 2 (Sh2)-Containing Proteins Near The Plasma Membrane, Dongmyung Oh, Mari Ogiue-Ikeda, Joshua A. Jadwin, Kazuya Machida, Bruce J. Mayer, Ji Yu

UCHC Articles - Research

Receptor tyrosine kinases (RTKs) control a host of biological functions by phosphorylating tyrosine residues of intracellular proteins upon extracellular ligand binding. The phosphotyrosines (p-Tyr) then recruit a subset of ∼100 Src homology 2 (SH2) domain-containing proteins to the cell membrane. The in vivo kinetics of this process are not well understood. Here we use total internal reflection (TIR) microscopy and single-molecule imaging to monitor interactions between SH2 modules and p-Tyr sites near the cell membrane. We found that the dwell time of SH2 modules within the TIR illumination field is significantly longer than predictions based on chemical dissociation rate constants, …

Family-Based Genome-Wide Association Study Of Frontal Theta Oscillations Identifies Potassium Channel Gene Kcnj6, Victor M. Hesselbrock

UCHC Articles - Research

Event-related oscillations (EROs) represent highly heritable neuroelectric correlates of cognitive processes that manifest deficits in alcoholics and in offspring at high risk to develop alcoholism. Theta ERO to targets in the visual oddball task has been shown to be an endophenotype for alcoholism. A family-based genome-wide association study was performed for the frontal theta ERO phenotype using 634583 autosomal single nucleotide polymorphisms (SNPs) genotyped in 1560 family members from 117 families densely affected by alcohol use disorders, recruited in the Collaborative Study on the Genetics of Alcoholism. Genome-wide significant association was found with several SNPs on chromosome 21 in KCNJ6 …

Interleukin-1Β In Central Nervous System Injury And Repair, Nicole A. Jackman, Sandra J. Hewett, Robert J. Claycomb

UCHC Articles - Research

Acute inflammation is a self-limiting, complex biological response mounted to combat pathogen invasion, to protect against tissue damage, and to promote tissue repair should it occur. However, unabated inflammation can be deleterious and contribute to injury and pathology. Interleukin-1β (IL-1β), a prototypical “pro-inflammatory” cytokine, is essential to cellular defense and tissue repair in nearly all tissues. With respect to brain, however, studies suggest that IL-1β has pleiotrophic effects. It acts as a neuromodulator in the healthy central nervous system (CNS), has been implicated in the pathogenic processes associated with a number of CNS maladies, but may also provide protection to …

Spontaneous Spheroid Budding From Monolayers: A Potential Contribution To Ovarian Cancer Dissemination, Jillian C. Pease, Molly Brewer, Jennifer S. Tirnauer

UCHC Articles - Research

Ovarian cancer is the most lethal gynaecologic cancer, in large part because of its early dissemination and rapid development of chemotherapy resistance. Spheroids are clusters of tumor cells found in the peritoneal fluid of patients that are thought to promote this dissemination. Current models suggest that spheroids form by aggregation of single tumor cells shed from the primary tumor. Here, we demonstrate that spheroids can also form by budding directly as adherent clusters from a monolayer. Formation of budded spheroids correlated with expression of vimentin and lack of cortical E-cadherin. We also found that compared to cells grown in monolayers, …

Cytochrome B5 And Cytokeratin 17 Are Biomarkers In Bronchoalveolar Fluid Signifying Onset Of Acute Lung Injury, Antoine Ménoret, Sanjeev Kumar, Anthony T. Vella

UCHC Articles - Research

Acute lung injury (ALI) is characterized by pulmonary edema and acute inflammation leading to pulmonary dysfunction and potentially death. Early medical intervention may ameliorate the severity of ALI, but unfortunately, there are no reliable biomarkers for early diagnosis. We screened for biomarkers in a mouse model of ALI. In this model, inhalation of S. aureus enterotoxin A causes increased capillary permeability, cell damage, and increase protein and cytokine concentration in the lungs. We set out to find predictive biomarkers of ALI in bronchoalveolar lavage (BAL) fluid before the onset of clinical manifestations. A cutting edge proteomic approach was used to …

Current And Emerging Therapies In Multiple Sclerosis: A Systematic Review, Wanda Castro-Borrero

UCHC Articles - Research

Multiple sclerosis (MS) is a potentially disabling chronic autoimmune neurological disease that mainly affects young adults. Our understanding of the pathophysiology of MS has significantly advanced in the past quarter of a century. This has led to the development of many disease-modifying therapies (DMTs) that prevent exacerbations and new lesions in patients with relapsing remitting MS (RRMS). So far there is no drug available that can completely halt the neurodegenerative changes associated with the disease. It is the purpose of this review to provide concise information regarding mechanism of action, indications, side effects and safety of Food and Drug Administration …

Determining The Functional Significance Of Mismatch Repair Gene Missense Variants Using Biochemical And Cellular Assays, Christopher D. Heinen

UCHC Articles - Research

With the discovery that the hereditary cancer susceptibility disease Lynch syndrome (LS) is caused by deleterious germline mutations in the DNA mismatch repair (MMR) genes nearly 20 years ago, genetic testing can now be used to diagnose this disorder in patients. A definitive diagnosis of LS can direct how clinicians manage the disease as well as prevent future cancers for the patient and their families. A challenge emerges, however, when a germline missense variant is identified in a MMR gene in a suspected LS patient. The significance of a single amino acid change in these large repair proteins is not …

Reaction-Diffusion Modeling In The Neuron Simulator, James C. Schaff

UCHC Articles - Research

The NEURON simulator is a widely used tool for studying detailed single cell and network models. In recognition of the growing importance of multi-scale modeling, we have expanded NEURON’s support for intracellular chemical dynamics. In particular, we discuss our work with stochastic reaction-diffusion models and three-dimensional simulations.

High Serum Cu And Cu/Zn Ratios Correlate With Impairments In Bone Density, Physical Performance And Overall Health In A Population Of Elderly Men With Frailty Characteristics, Eric D. Gaier, Alison Kleppinger, Richard E. Mains, Anne M. Kenny, Betty A. Eipper

UCHC Articles - Research

Serum Cu levels rise with age and high Cu/Zn ratios are linked with multiple-cause mortality in the elderly. The relationships of these parameters to measures of musculoskeletal health and frailty have not yet been analyzed. We used inductively coupled mass spectrometry to assess serum levels of Cu and Zn and probed for relationships between serum Cu levels and the Cu/Zn ratio with specific measures of bone, physical and overall health in a cohort of 144 frail elderly men. Subjects were divided into quintiles based on serum metal levels and comparisons for functional measures were made between the reference (middle) group …

Computational Analysis Of Calcium Signaling And Membrane Electrophysiology In Cerebellar Purkinje Neurons Associated With Ataxia, Leslie M. Loew, Sherry-Ann Brown

UCHC Articles - Research

Background

Mutations in the smooth endoplasmic reticulum (sER) calcium channel Inositol Trisphosphate Receptor type 1 (IP3R1) in humans with the motor function coordination disorders Spinocerebellar Ataxia Types 15 and 16 (SCA15/16) and in a corresponding mouse model, the IP3R1delta18/delta18 mice, lead to reduced IP3R1 levels. We posit that increasing IP3R1 sensitivity to IP3 in ataxias with reduced IP3R1 could restore normal calcium response. On the other hand, in mouse models of the human polyglutamine (polyQ) ataxias, SCA2, and SCA3, the primary finding appears to be hyperactive IP3R1-mediated calcium release. It has been suggested that the polyQ SCA1 mice may also …

A Gutsy Way To Extend Longevity, Ryan P. Rogers, Blanka Rogina

UCHC Articles - Research

The studies conducted by Rera et al. successfully demonstrate a new role for dPGC-1 and mitochondrial biogenesis in ISC homeostasis and longevity. How does it all work? dPGC-1 overexpression increases levels of anti-oxidative enzymes to decrease oxidative damage and conserve ISC homeostasis. Preserved ISC homeostasis is required for gut maintenance, which is necessary for normal energy supply and/or prevention of microbial or toxin overload. All of these factors may contribute to delaying the onset of age related phenotypes. As Zhou et al. (2011) suggested, it is also possible that an unknown factor/s released by PGC-1 overexpression in ISCs/EBs regulates longevity …

Global Approaches To The Role Of Mirnas In Drug-Induced Changes In Gene Expression, Jodi E. Eipper-Mains, Betty A. Eipper, Richard E. Mains

UCHC Articles - Research

Neurons modulate gene expression with subcellular precision through excitation-coupled local protein synthesis, a process that is regulated in part through the involvement of microRNAs (miRNAs), a class of small non-coding RNAs. The biosynthesis of miRNAs is reviewed, with special emphasis on miRNA families, the subcellular localization of specific miRNAs in neurons, and their potential roles in the response to drugs of abuse. For over a decade, DNA microarrays have dominated genome-wide gene expression studies, revealing widespread effects of drug exposure on neuronal gene expression. We review a number of recent studies that explore the emerging role of miRNAs in the …

Approaches To The Study Of Neural Coding Of Sound Source Location And Sound Envelope In Real Environments, Shigeyuki Kuwada, Brian Bishop, Duck O. Kim

UCHC Articles - Research

The major functions of the auditory system are recognition (what is the sound) and localization (where is the sound). Although each of these has received considerable attention, rarely are they studied in combination. Furthermore, the stimuli used in the bulk of studies did not represent sound location in real environments and ignored the effects of reverberation. Another ignored dimension is the distance of a sound source. Finally, there is a scarcity of studies conducted in unanesthetized animals. We illustrate a set of efficient methods that overcome these shortcomings. We use the virtual auditory space method (VAS) to efficiently present sounds …

Model Selection For Cox Models With Time-Varying Coefficients, Jun Yan

UCHC Articles - Research

Cox models with time-varying coefficients offer great flexibility in capturing the temporal dynamics of covariate effects on right censored failure times. Since not all covariate coefficients are time-varying, model selection for such models presents an additional challenge, which is to distinguish covariates with time-varying coefficient from those with time-independent coefficient. We propose an adaptive group lasso method that not only selects important variables but also selects between time-independent and time-varying specifications of their presence in the model. Each covariate effect is partitioned into a time-independent part and a time-varying part, the latter of which is characterized by a group of …

Loss Of Vascular Early Response Gene Reduces Edema Formation After Experimental Stroke, Fudong Liu, L C. Turtzo, Jun Li, Neer Zeevi, Louise D. Mccullough

UCHC Articles - Research

Vascular Early Response Gene (Verge) is an immediate early gene (IEG) that is up-regulated in endothelial cells in response to a number of stressors, including ischemic stroke. Endothelial cell lines that stably express Verge show enhanced permeability. Increased Verge expression has also been associated with blood brain barrier breakdown. In this study we investigated the role of Verge in ischemic injury induced by middle cerebral artery occlusion (MCAO) in both Verge knockout (KO) and wild type (WT) mice. Verge KO mice had significantly less cerebral edema formation after MCAO compared to WT mice. However, stroke outcome (infarct size and neurological …

Genomic Analysis Of Pterostilbene Predicts Its Antiproliferative Effects Against Pancreatic Cancer In Vitro And In Vivo, David W. Mcfadden

UCHC Articles - Research

Background

To investigate the inhibitory role of pterostilbene in pancreatic cancer, we conducted a genomic analysis of pterostilbene-treated pancreatic cancer cells. We also investigated the effect of pterostilbene upon the carcinogenic markers, manganese superoxide dismutase, cytochrome C, Smac/DIABLO, and STAT3 phosphorylation in vitro. The antiproliferative effects of pterostilbene were further evaluated in an in vivo model.

Methods

Pancreatic cancer cells were treated with pterostilbene and evaluated with DNA microarray analysis. Pterostilbenetreated cells were analyzed for cytochrome C, Smac/DIABLO, manganese superoxide dismutase (MnSOD)/antioxidant activity, and STAT3 phosphorylation using ELISA. Data were statistically analyzed using ANOVA. Pterostilbene was then administered to nude …

Frequency Weighting For Vibration-Induced White Finger Compatible With Exposure-Response Models., Anthony J. Brammer

UCHC Articles - Research

An analysis has been performed to derive a frequency weighting for the development of vibration-induced white finger (VWF). It employs a model to compare health risks for pairs of population groups that are selected to have similar health outcomes from operating power tools or machines with markedly different acceleration spectra (rock drills, chain saws, pavement breakers and motorcycles). The model defines the Relative Risk, RR_f(trial), which is constructed from the ratio of daily exposures and includes a trial frequency weighting that is applied to the acceleration spectra. The trial frequency weighting consists of a frequency-independent primary frequency range, …