Digital Commons Network^™

Dual Engagement Of The Nlrp3 And Aim2 Inflammasomes By Plasmodium-Derived Hemozoin And Dna During Malaria, Parisa Kalantari, Rosane B. Deoliveira, Jennie Chan, Yolanda Corbett, Vijay A. K. Rathinam, Andrea Stutz, Eicke Latz, Ricardo T. Gazzinelli, Douglas T. Golenbock, Katherine A. Fitzgerald

Katherine A. Fitzgerald

Hemozoin (Hz) is the crystalline detoxification product of hemoglobin in Plasmodium-infected erythrocytes. We previously proposed that Hz can carry plasmodial DNA into a subcellular compartment that is accessible to Toll-like receptor 9 (TLR9), inducing an inflammatory signal. Hz also activates the NLRP3 inflammasome in primed cells. We found that Hz appears to colocalize with DNA in infected erythrocytes, even before RBC rupture or phagolysosomal digestion. Using synthetic Hz coated in vitro with plasmodial genomic DNA (gDNA) or CpG oligodeoxynucleotides, we observed that DNA-complexed Hz induced TLR9 translocation, providing a priming and an activation signal for inflammasomes. After phagocytosis, Hz and …

Nuclear Transport Of Single Molecules: Dwell Times At The Nuclear Pore Complex, Ulrich Kubitscheck, David Grunwald, Andreas Hoekstra, Daniel Rohleder, Thorsten Kues, Jan Peter Siebrasse, Reiner Peters

David Grünwald

The mechanism by which macromolecules are selectively translocated through the nuclear pore complex (NPC) is still essentially unresolved. Single molecule methods can provide unique information on topographic properties and kinetic processes of asynchronous supramolecular assemblies with excellent spatial and time resolution. Here, single-molecule far-field fluorescence microscopy was applied to the NPC of permeabilized cells. The nucleoporin Nup358 could be localized at a distance of 70 nm from POM121-GFP along the NPC axis. Binding sites of NTF2, the transport receptor of RanGDP, were observed in cytoplasmic filaments and central framework, but not nucleoplasmic filaments of the NPC. The dwell times of …

Intranuclear Binding Kinetics And Mobility Of Single Native U1 Snrnp Particles In Living Cells, David Grunwald, Beatrice Spottke, Volker Buschmann, Ulrich Kubitscheck

David Grünwald

Uridine-rich small nuclear ribonucleoproteins (U snRNPs) are splicing factors, which are diffusely distributed in the nucleoplasm and also concentrated in nuclear speckles. Fluorescently labeled, native U1 snRNPs were microinjected into the cytoplasm of living HeLa cells. After nuclear import single U1 snRNPs could be visualized and tracked at a spatial precision of 30 nm at a frame rate of 200 Hz employing a custom-built microscope with single-molecule sensitivity. The single-particle tracks revealed that most U1 snRNPs were bound to specific intranuclear sites, many of those presumably representing pre-mRNA splicing sites. The dissociation kinetics from these sites showed a multiexponential decay …

Conversion Of Red Fluorescent Protein Into A Bright Blue Probe, Oksana M. Subach, Illia S. Gundorov, Masami Yoshimura, Fedor V. Subach, Jinghang Zhang, David Grunwald, Ekaterina A. Souslova, Dmitriy M. Chudakov, Vladislav V. Verkhusha

David Grünwald

We used a red chromophore formation pathway, in which the anionic red chromophore is formed from the neutral blue intermediate, to suggest a rational design strategy to develop blue fluorescent proteins with a tyrosine-based chromophore. The strategy was applied to red fluorescent proteins of the different genetic backgrounds, such as TagRFP, mCherry, HcRed1, M355NA, and mKeima, which all were converted into blue probes. Further improvement of the blue variant of TagRFP by random mutagenesis resulted in an enhanced monomeric protein, mTagBFP, characterized by the substantially higher brightness, the faster chromophore maturation, and the higher pH stability than blue fluorescent proteins …

Autonomy And Robustness Of Translocation Through The Nuclear Pore Complex: A Single-Molecule Study, Thomas Dange, David Grunwald, Antje Grunwald, Reiner Peters, Ulrich Kubitscheck

David Grünwald

All molecular traffic between nucleus and cytoplasm occurs via the nuclear pore complex (NPC) within the nuclear envelope. In this study we analyzed the interactions of the nuclear transport receptors kapalpha2, kapbeta1, kapbeta1DeltaN44, and kapbeta2, and the model transport substrate, BSA-NLS, with NPCs to determine binding sites and kinetics using single-molecule microscopy in living cells. Recombinant transport receptors and BSA-NLS were fluorescently labeled by AlexaFluor 488, and microinjected into the cytoplasm of living HeLa cells expressing POM121-GFP as a nuclear pore marker. After bleaching the dominant GFP fluorescence the interactions of the microinjected molecules could be studied using video microscopy …

Nuclear Pore Component Nup98 Is A Potential Tumor Suppressor And Regulates Posttranscriptional Expression Of Select P53 Target Genes, Stephan Singer, Ruiying Zhao, Anthony M. Barsotti, Anette Ouwehand, Mina Fazollahi, Elias Coutavas, Kai Breuhahn, Olaf Neumann, Thomas Longerich, Tobias Pusterla, Maureen A. Powers, Keith M. Giles, Peter J. Leedman, Jochen Hess, David Grunwald, Harmen J. Bussemaker, Robert H. Singer, Peter Schirmacher, Carol Prives

David Grünwald

The p53 tumor suppressor utilizes multiple mechanisms to selectively regulate its myriad target genes, which in turn mediate diverse cellular processes. Here, using conventional and single-molecule mRNA analyses, we demonstrate that the nucleoporin Nup98 is required for full expression of p21, a key effector of the p53 pathway, but not several other p53 target genes. Nup98 regulates p21 mRNA levels by a posttranscriptional mechanism in which a complex containing Nup98 and the p21 mRNA 3'UTR protects p21 mRNA from degradation by the exosome. An in silico approach revealed another p53 target (14-3-3sigma) to be similarly regulated by Nup98. The expression …

Imp3 Expression Is Associated With Poor Outcome And Epigenetic Deregulation In Intrahepatic Cholangiocarcinoma, Yuanyuan Gao, Michelle Yang, Zhong Jiang, Bruce A. Woda, Arthur M. Mercurio, Jianjie Qin, Xinli Huang, Feng Zhang

Arthur M. Mercurio

IMP3 is a fetal protein not expressed in normal adult tissues. IMP3 is an oncoprotein and a useful biomarker for a variety of malignancies and is associated with reduced overall survival of a number of them. IMP3 expression and its prognostic value for patients with intrahepatic cholangiocarcinoma (ICC) have not been well investigated. The molecular mechanism underlying IMP3 expression in human cancer cells remains to be elucidated. Here we investigated IMP3 expression in ICC and adjacent nonneoplastic liver in 72 unifocal primary ICCs from a single institute by immunohistochemistry, immunoblotting, and real-time polymerase chain reaction. IMP3 was specifically expressed in …

Temporal Trends In Serum Concentrations Of Polychlorinated Dioxins, Furans, And Pcbs Among Adult Women Living In Chapaevsk, Russia: A Longitudinal Study From 2000 To 2009, Olivier Humblet, Oleg Sergeyev, Larisa Altshul, Susan A. Korrick, Paige L. Williams, Claude Emond, Linda S. Birnbaum, Jane S. Burns, Mary M. Lee, Boris Revich, Andrey Shelepchikov, Denis Feshin, Russ Hauser

Mary M. Lee

BACKGROUND: The present study assessed the temporal trend in serum concentrations of polychlorinated dibenzo-p-dioxins, dibenzofurans, and biphenyls (PCBs) among residents of a Russian town where levels of these chemicals are elevated due to prior industrial activity.

METHODS: Two serum samples were collected from eight adult women (in 2000 and 2009), and analyzed with gas chromatography-high-resolution mass spectrometry.

RESULTS: The average total toxic equivalency (TEQ) decreased by 30% (from 36 to 25 pg/g lipid), and the average sum of PCB congeners decreased by 19% (from 291 to 211 ng/g lipid). Total TEQs decreased for seven of the eight women, and the …

Predictors Of Serum Dioxins And Pcbs Among Peripubertal Russian Boys, Jane S. Burns, Paige L. Williams, Oleg Sergeyev, Susan Korrick, Mary M. Lee, Boris Revich, Larisa Altshul, Donald G. Patterson, Wayman E. Turner, Larry L. Needham, Igor Saharov, Russ Hauser

Mary M. Lee

BACKGROUND: Although sources and routes of exposure to dioxins and polychlorinated biphenyls (PCBs) have been studied, information regarding exposure among children is limited. Breast-feeding and diet are two important contributors to early life exposure. To further understand other significant contributors to childhood exposure, we studied a cohort of children from a city with high environmental dioxin levels. OBJECTIVES: We investigated predictors of serum concentrations of polychlorinated dibenzo-p-dioxins (PCDDs)/polychlorinated dibenzofurans (PCDFs)/co-planar PCBs (C-PCBs), toxic equivalents (TEQs), and PCBs among 8- to 9-year-old boys in Chapaevsk, Russia. METHODS: We used general linear regression models to explore associations of log(10)-transformed serum concentrations of …

Association Of Blood Lead Levels With Onset Of Puberty In Russian Boys, Russ Hauser, Oleg Sergeyev, Susan Korrick, Mary M. Lee, Boris Revich, Elana Gitin, Jane S. Burns, Paige L. Williams

Mary M. Lee

BACKGROUND: Epidemiologic studies suggest a temporal trend of earlier onset and longer duration of puberty, raising concerns regarding the potential impact of environmental factors on pubertal development. Lead exposure has been associated with delayed pubertal onset in girls; however, epidemiologic data in boys are limited. METHODS: We used multivariable logistic regression models to explore the cross-sectional association of blood lead levels with growth and pubertal onset based on physician-assessed testicular volume (TV) and pubertal staging in 489 boys 8-9 years of age from Chapaevsk, Russia. We used multivariable linear regression models to assess associations of blood lead levels with somatic …

Blood Lead Levels And Serum Insulin-Like Growth Factor 1 Concentrations In Peripubertal Boys, Abby F. Fleisch, Jane S. Burns, Paige L. Williams, Mary M. Lee, Oleg Sergeyev, Susan Korrick, Russ Hauser

Mary M. Lee

BACKGROUND: Childhood lead exposure has been associated with growth delay. However, the association between blood lead levels (BLLs) and insulin-like growth factor 1 (IGF-1) has not been characterized in a large cohort with low-level lead exposure.

METHODS: We recruited 394 boys 8-9 years of age from an industrial Russian town in 2003-2005 and followed them annually thereafter. We used linear regression models to estimate the association of baseline BLLs with serum IGF-1 concentration at two follow-up visits (ages 10-11 and 12-13 years), adjusting for demographic and socioeconomic covariates.

RESULTS: At study entry, median BLL was 3 mug/dL (range, < 0.5-31 mug/dL), most boys (86%) were prepubertal, and mean +/- SD height and BMI z-scores were 0.14 +/- 1.0 and -0.2 +/- 1.3, respectively. After adjustment for covariates, the mean follow-up IGF-1 concentration was 29.2 ng/mL lower (95% CI: -43.8, -14.5) for boys with high versus low BLL (>/= 5 …

Predictors Of Serum Dioxin Levels Among Adolescent Boys In Chapaevsk, Russia: A Cross-Sectional Pilot Study, Russ Hauser, Paige L. Williams, Larisa Altshul, Susan Korrick, Lynne Peeples, Donald G. Patterson Jr., Wayman E. Turner, Mary M. Lee, Boris Revich, Oleg Sergeyev

Mary M. Lee

BACKGROUND: Toxicological studies and limited human studies have demonstrated associations between exposure to polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and polychlorinated biphenyls (PCBs) and adverse developmental and reproductive health effects. Given that children may be particularly susceptible to reproductive and developmental effects of organochlorines, and the paucity of information available regarding childhood exposures to dioxins in particular, we undertook a pilot study to describe the distribution of, and identify potential predictors of exposure to, dioxin-like compounds and dioxins among adolescent boys in Chapaevsk, Russia. The pilot study was also designed to guide the development of a large prospective cohort study …

Parental Mastery Of Continuous Subcutaneous Insulin Infusion Skills And Glycemic Control In Youth With Type 1 Diabetes, Kathleen Mitchell, Kimberley Johnson, Karen Cullen, Mary M. Lee, Olga T. Hardy

Mary M. Lee

OBJECTIVE: The purpose of this study is to determine whether parental knowledge of the continuous subcutaneous insulin infusion (CSII) device affects glycemic control as measured by hemoglobin A1c (A1C) level. SUBJECTS AND METHODS: Parents of children with type 1 diabetes mellitus (T1DM) using CSII completed a 14-item questionnaire. Questions 1-10 were knowledge-based questions that required the parent to extract specific information from their child's CSII device. Questions 11-14 asked parents to provide a self-assessment of their CSII knowledge. RESULTS: Twenty-two parents of youth with T1DM participated in the study. Ten of the youth were in the Low-A1C group (A1C/=8%). Parents …

Hsisp Annotated Bibliography: Attitudes Toward Animals (1998-2013), Erich Yahner

Erich Yahner, MSLIS

No abstract provided.

Dengue Virus Nonstructural Protein Ns5 Induces Interleukin-8 Transcription And Secretion, Carey L. Medin, Katherine A. Fitzgerald, Alan L. Rothman

Alan Rothman

Elevated circulating levels of chemokines have been reported in patients with dengue fever and are proposed to contribute to the pathogenesis of dengue disease. To establish in vitro models for chemokine induction by dengue 2 virus (DEN2V), we studied a variety of human cell lines and primary cells. DEN2V infection of HepG2 and primary dendritic cells induced the production of interleukin-8 (IL-8), RANTES, MIP-1alpha, and MIP-1beta, whereas only IL-8 and RANTES were induced following dengue virus infection of HEK293 cells. Chemokine secretion was accompanied by an increase in steady-state mRNA levels. No chemokine induction was observed in HEK293 cells treated …

A Pilot Study Of Qigong For Reducing Cocaine Craving Early In Recovery, David A. Smelson, Kevin W. Chen, Douglas M. Ziedonis, Ken Andes, Amanda Lennox, Lanora Callahan, Stephanie Rodrigues, David Eisenberg

Douglas M. Ziedonis

OBJECTIVES: This pilot study examined the feasibility, preliminary efficacy, and determined the effect sizes of external qigong therapy (EQT) in reducing cue-elicited cocaine craving and associated symptoms among recently abstinent cocaine-dependent (CD) individuals. METHODS: This study randomized 101 CD subjects to either a real EQT (n=51) or sham EQT control (n=50) group. Subjects underwent a baseline assessment and a weekly cue-exposure session for 2 weeks. Total EQT or sham treatments ranged from 4 to 6 sessions in 2 weeks. RESULTS: EQT-treated subjects displayed a greater reduction in cue-elicited craving (p=0.06) and symptoms of depression (p<0.05) with medium effect sizes. …

Test Order Effects In Simultaneous Protocols, Abdulrazaq Imam, Timothy A. Warner

Abdulrazaq A. Imam

Simultaneous protocols typically yield poorer stimulus equivalence outcomes than do other protocols commonly used in equivalence research. Two independent groups of three 3-member equivalence sets of stimuli were used in conditional discrimination procedures in two conditions, one using the standard simultaneous protocol and the other using a hybrid simultaneous training and simple-to-complex testing. Participants completed the two conditions in one long session in Experiment 1, but in separate sessions in Experiment 2. The same stimulus sets used in Experiment 1 were randomized for the two conditions in Experiment 2. Overall, accuracy was better with the hybrid than with the standard …

Can Exercise Ameliorate Treatment Toxicity During The Initial Phase Of Testosterone Deprivation In Prostate Cancer Patients? Is This More Effective Than Delayed Rehabilitation?, Robert Newton, Dennis Taaffe, Nigel Spry, Prue Cormie, Suzanne Chambers, Robert Gardiner, David Shum, David Joseph, Daniel Galvao

Prue Cormie

Background: There has been substantial increase in use of androgen deprivation therapy as adjuvant management of prostate cancer. However, this leads to a range of musculoskeletal toxicities including reduced bone mass and increased skeletal fractures compounded with rapid metabolic alterations, including increased body fat, reduced lean mass, insulin resistance and negative lipoprotein profile, increased incidence of cardiovascular and metabolic morbidity, greater distress and reduced quality of life. Numerous research studies have demonstrated certain exercise prescriptions to be effective at preventing or even reversing these treatment toxicities. However, all interventions to date have been of rehabilitative intent being implemented after a …

The Future Prospects Of Embedded Microchips In Humans As Unique Identifiers: The Risks Versus The Rewards, Katina Michael, M G. Michael

Professor Katina Michael

Microchip implants for humans are not new. Placing heart pacemakers in humans for prosthesis is now considered a straightforward procedure. In more recent times we have begun to use brain pacemakers for therapeutic purposes to combat illnesses such as epilepsy, Parkinson's disease, and severe depression. Microchips are even being placed inside prosthetic knees and hips during restorative procedures to help in the gathering of post-operative analytics that can aid rehabilitation further. While medical innovations that utilize microchips abound, over the last decade we have begun to see the potential use of microchip implants for non-medical devices in humans, namely for …

Effect Of The Planet Health Intervention On Eating Disorder Symptoms In Massachusetts Middle Schools, 2005-2008, S. Bryn Austin, Jennifer L. Spadano-Gasbarro, Mary L. Greaney, Emily A. Blood, Anne T. Hunt, Tracy K. Richmond, Monica L. Wang, Solomon Mezgebu, Stavroula K. Osganian, Karen E. Peterson

Monica L. Wang

INTRODUCTION: The Planet Health obesity prevention curriculum has prevented purging and abuse of diet pills (disordered weight control behavior [DWCB]) in middle-school girls in randomized trials, but the effects of Planet Health on DWCB when implemented by schools under dissemination conditions are not known. METHODS: Massachusetts Department of Public Health and Blue Cross Blue Shield of Massachusetts disseminated Planet Health as part of the 3-year, Healthy Choices obesity prevention program in middle schools. We conducted an evaluation in 45 schools from fall 2005 to spring 2008. We gathered data from school staff to quantify intervention activities, and we gathered anonymous …

Tom Regan On ‘Kind’ Arguments Against Animal Rights And For Human Rights, Nathan Nobis

Nathan M. Nobis, PhD

Prenatal Oral Health Education In U.S. Dental Schools And Obstetrics And Gynecology Residencies, Megan Curtis, Hugh Silk, Judith A. Savageau

Judith A. Savageau

Prenatal oral health (POH) is an important health issue, but dental and obstetrical clinicians are not meeting the oral health needs of pregnant patients. This study evaluates how training contributes to this paradox with a national survey of sixty dental school deans and 240 obstetrics and gynecology residency program directors. Response rates were 53 percent and 40 percent for deans and program directors, respectively. According to the respondents, 94 percent of responding dental schools provided POH education, only 39 percent of responding residencies taught POH, and 65 percent of responding deans and 45 percent of responding program directors were aware …

Determinants Of Inapparent And Symptomatic Dengue Infection In A Prospective Study Of Primary School Children In Kamphaeng Phet, Thailand, Timothy P. Endy, Kathryn B. Anderson, Ananda Nisalak, In-Kyu Yoon, Sharone Green, Alan L. Rothman, Stephen J. Thomas, Richard G. Jarman, Daniel H. Libraty, Robert V. Gibbons

Sharone Green

BACKGROUND: Dengue viruses are a major cause of morbidity in tropical and subtropical regions of the world. Inapparent dengue is an important component of the overall burden of dengue infection. It provides a source of infection for mosquito transmission during the course of an epidemic, yet by definition is undetected by health care providers. Previous studies of inapparent or subclinical infection have reported varying ratios of symptomatic to inapparent dengue infection. METHODOLOGY/PRINCIPAL FINDINGS: In a prospective study of school children in Northern Thailand, we describe the spatial and temporal variation of the symptomatic to inapparent (S:I) dengue illness ratio. Our …

T Cell Receptor Vbeta Gene Usage In Thai Children With Dengue Virus Infection, Susan J. Gagnon, Anita M. Leporati, Sharone Green, Siripen Kalayanarooj, David W. Vaughn, Henry A. F. Stephens, Saroj Suntayakorn, Ichiro Kurane, Francis A. Ennis, Alan L. Rothman

Sharone Green

T lymphocyte activation during dengue is thought to contribute to the pathogenesis of dengue hemorrhagic fever (DHF). We examined the T cell receptor Vbeta gene usage by a reverse transcriptase-polymerase chain reaction assay during infection and after recovery in 13 children with DHF and 13 children with dengue fever (DF). There was no deletion of specific Vbeta gene families. We detected significant expansions in usage of single Vbeta families in six subjects with DHF and three subjects with DF over the course of infection, but these did not show an association with clinical diagnosis, viral serotype, or HLA alleles. Differences …

Prediction Of Dengue Disease Severity Among Pediatric Thai Patients Using Early Clinical Laboratory Indicators, James A. Potts, Robert V. Gibbons, Alan L. Rothman, Anon Srikiatkhachorn, Stephen J. Thomas, Pra-On Supradish, Stephenie C. Lemon, Daniel H. Libraty, Sharone Green, Siripen Kalayanarooj

Sharone Green

BACKGROUND: Dengue virus is endemic in tropical and sub-tropical resource-poor countries. Dengue illness can range from a nonspecific febrile illness to a severe disease, Dengue Shock Syndrome (DSS), in which patients develop circulatory failure. Earlier diagnosis of severe dengue illnesses would have a substantial impact on the allocation of health resources in endemic countries. METHODS AND FINDINGS: We compared clinical laboratory findings collected within 72 hours of fever onset from a prospective cohort children presenting to one of two hospitals (one urban and one rural) in Thailand. Classification and regression tree analysis was used to develop diagnostic algorithms using different …

Virus-Induced Decline In Soluble Vascular Endothelial Growth Receptor 2 Is Associated With Plasma Leakage In Dengue Hemorrhagic Fever, Anon Srikiatkhachorn, Chuanpis Ajariyakhajorn, Timothy P. Endy, Siripen Kalayanarooj, Daniel H. Libraty, Sharone Green, Francis A. Ennis, Alan L. Rothman

Sharone Green

Some individuals infected with dengue virus develop dengue hemorrhagic fever (DHF), a viral hemorrhagic disease characterized by a transient period of localized plasma leakage. To determine the importance of vascular endothelial growth factor A (VEGF-A) in this syndrome, we compared plasma levels of VEGF-A and the soluble forms of its receptors in patients with DHF to patients with dengue fever (DF), a milder form of dengue virus infection without plasma leakage. We observed a rise in the plasma levels of free, but not total VEGF-A in DHF patients at the time of plasma leakage. This was associated with a decline …

The Future Prospects Of Embedded Microchips In Humans As Unique Identifiers: The Risks Versus The Rewards, Katina Michael, M G. Michael

M. G. Michael

Microchip implants for humans are not new. Placing heart pacemakers in humans for prosthesis is now considered a straightforward procedure. In more recent times we have begun to use brain pacemakers for therapeutic purposes to combat illnesses such as epilepsy, Parkinson's disease, and severe depression. Microchips are even being placed inside prosthetic knees and hips during restorative procedures to help in the gathering of post-operative analytics that can aid rehabilitation further. While medical innovations that utilize microchips abound, over the last decade we have begun to see the potential use of microchip implants for non-medical devices in humans, namely for …