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Improved Adenovirus Type 5 Vector-Mediated Transduction of Resistant Cells by Piggybacking on Coxsackie B-Adenovirus Receptor-Pseudotyped Baculovirus?  

PubMed Central

Taking advantage of the wide tropism of baculoviruses (BVs), we constructed a recombinant BV (BVCAR) pseudotyped with human coxsackie B-adenovirus receptor (CAR), the high-affinity attachment receptor for adenovirus type 5 (Ad5), and used the strategy of piggybacking Ad5-green fluorescent protein (Ad5GFP) vector on BVCAR to transduce various cells refractory to Ad5 infection. We found that transduction of all cells tested, including human primary cells and cancer cell lines, was significantly improved using the BVCAR-Ad5GFP biviral complex compared to that obtained with Ad5GFP or BVCARGFP alone. We determined the optimal conditions for the formation of the complex and found that a high level of BVCAR-Ad5GFP-mediated transduction occurred at relatively low adenovirus vector doses, compared with transduction by Ad5GFP alone. The increase in transduction was dependent on the direct coupling of BVCAR to Ad5GFP via CAR-fiber knob interaction, and the cell attachment of the BVCAR-Ad5GFP complex was mediated by the baculoviral envelope glycoprotein gp64. Analysis of the virus-cell binding reaction indicated that the presence of BVCAR in the complex provided kinetic benefits to Ad5GFP compared to the effects with Ad5GFP alone. The endocytic pathway of BVCAR-Ad5GFP did not require Ad5 penton base RGD-integrin interaction. Biodistribution of BVCAR-Ad5Luc complex in vivo was studied by intravenous administration to nude BALB/c mice and compared to Ad5Luc injected alone. No significant difference in viscerotropism was found between the two inocula, and the liver remained the preferred localization. In vitro, coagulation factor X drastically increased the Ad5GFP-mediated transduction of CAR-negative cells but had no effect on the efficiency of transduction by the BVCAR-Ad5GFP complex. Various situations in vitro or ex vivo in which our BVCAR-Ad5 duo could be advantageously used as gene transfer biviral vector are discussed. PMID:19357170

Granio, Ophelia; Porcherot, Marine; Corjon, Stephanie; Kitidee, Kuntida; Henning, Petra; Eljaafari, Assia; Cimarelli, Andrea; Lindholm, Leif; Miossec, Pierre; Boulanger, Pierre; Hong, Saw-See



Myocardial imaging. Coxsackie myocarditis  

SciTech Connect

A 3-week-old male neonate with heart failure associated with Coxsackie virus infection was imaged with Tc-99m PYP and TI-201. The abnormal imaging pattern suggested myocardial infarction. Autopsy findings indicated that the cause was myocardial necrosis secondary to an acute inflammatory process. Causes of abnormal myocardial uptake of Tc-99m PYP in pediatrics include infarction, myocarditis, cardiomyopathy, bacterial endocarditis, and trauma. Myocardial imaging cannot provide a specific cause diagnosis. Causes of myocardial infarction in pediatrics are listed in Table 1.

Wells, R.G.; Ruskin, J.A.; Sty, J.R.



136. Vector host interactions of group B adenoviruses in CD46 transgenic mice  

Microsoft Academic Search

Adenovirus (Ad) is a useful tool for gene delivery into a wide variety of cell types. Most adenoviral vectors used for gene therapy are based on serotype 5, which binds to cells via interactions between the viral fiber protein and its receptor, called Coxsackie and Adenovirus Receptor (CAR). Despite the widespread use of Ad5-based vectors for gene delivery, many therapeutic

Anuj Gaggar; Dmitry M. Shayakhmetov; Sonia Marwah; Shaoheng Ni; Andre M. Lieber



Model generation of viral channel forming 2B protein bundles from polio and coxsackie viruses  

E-print Network

Model generation of viral channel forming 2B protein bundles from polio and coxsackie viruses by enteroviruses such as polio and coxsackie viruses with two transmembrane domains. The protein is found to make this has on the in vivo activity of 2B. Keywords: 2B, polio virus, coxsackie virus, membrane proteins

Watts, Anthony


Reduced coxsackie antibody titres in Type 1 (insulin-dependent) diabetic patients presenting during an outbreak of coxsackie B 3 and B 4 infection  

Microsoft Academic Search

Summary  The potential role of antecedent viral infection in the pathogenesis of Type 1 (insulin-dependent) diabetes was investigated by measuring antibody titres to several viruses in serum obtained at the time of diagnosis of diabetes. An outbreak of Coxsackie B 4 infection folio wed by a wave of Coxsackie B 3 and B 5 infections occurred in Seattle during the time

J. P. Palmer; M. K. Cooney; R. H. Ward; J. A. Hansen; J. B. Brodsky; C. G. Ray; J. R. Crossley; C. M. Asplin; R. H. Williams



Susceptibility of Skeletal Muscle to Coxsackie A2 Virus Infection: Effects of Botulinum Toxin and Denervation  

NASA Astrophysics Data System (ADS)

Coxsackie A viruses can infect denervated but not innervated mature skeletal muscles. The role of synaptic transmission in preventing susceptibility to Coxsackievirus infection was studied by surgically denervating leg muscles of mice or injecting the muscles with botulinum toxin to block quantal release of acetylcholine. Control muscles were injected with heat-inactivated toxin. Subsequent injection of Coxsackie A2 virus resulted in extensive virus replication and tissue destruction in the denervated and botulinum toxin-treated muscles, while the control muscles showed only minimal changes. This suggests that the susceptibility of skeletal muscle to Coxsackievirus infection is regulated by synaptic transmission.

Andrew, Clifford G.; Drachman, Daniel B.; Pestronk, Alan; Narayan, Opendra



A study on dissociation of Coxsackie B4 virus-antibody complex  

Microsoft Academic Search

Dissociation of neutralizing antibody from Coxsackie B4 virus was achieved by acid treatment; pH 2.5 was found to be optimal for the reaction. Reactivated virus could be reneutralized by the dissociated antibody, but at a slower rate than the initial neutralization. Simple dilution was an effective method for recovering infectious virus from virus-antibody mixtures only if these mixtures were first

F Pinheiro; F. Hsiung




EPA Science Inventory

A seroepidemiological study was conducted to measure the antibody prevalence for eight different enteric viruses. These include seven 'classical' enteroviruses, ie, Coxsackie virus types A9, B1, B3, B4 and three ECHO virus types 4,7, and 9, as well as hepatitis A virus (HAV), rec...


The dual impact of coxsackie and adenovirus receptor expression on human prostate cancer gene therapy.  


In a recent paper, we reported a significant difference in coxsackie and adenovirus receptor (CAR) from several human bladder cancer cell lines that correlated with their sensitivities to adenoviral infection (Y. Li, R-C. Pong, J. M. Bergelson, M. C. Hall, A. I. Sagalowsky, C-P. Tseng, Z. Wang, and J. T. Hsieh, Cancer Res., 59: 325-330, 1999). In human prostate cancer, CAR protein is down-regulated in the highly tumorigenic PC3 cell line, which suggests that, in addition to its function as a viral receptor, CAR may have a pathophysiological role in prostate cancer progression. In this paper, we document that CAR does not merely enhance the viral sensitivity of prostate cancer cells but also acts as a tumor inhibitor for androgen-independent prostate cancer cells. Our data indicate that CAR is a potential therapeutic agent for increasing the efficacy of prostate cancer therapy. PMID:11016624

Okegawa, T; Li, Y; Pong, R C; Bergelson, J M; Zhou, J; Hsieh, J T



MDA5 and MAVS Mediate Type I Interferon Responses to Coxsackie B Virus?  

PubMed Central

Coxsackie B viruses (CVB) are enteroviruses that have been associated with a variety of human diseases, including myocarditis. In the present study, we found that MDA5 and its adaptor molecule MAVS are critical for type I interferon responses to CVB, since the absence of either MAVS or MDA5 leads to deficient type I interferon production and early mortality in mice infected with CVB. Pancreatic and hepatic necrosis were observed on histopathological examination of MAVS and MDA5 knockout mice infected with CVB. Inflammatory cytokine production in response to systemic CVB infection was independent of MAVS. Surprisingly, virus titers were not elevated in MAVS-deficient mice, despite significant reductions in type I interferon levels. These data highlight the importance of type I interferon in host defense and provide insight on the mechanisms of innate immune responses following coxsackievirus infection. PMID:19846534

Wang, Jennifer P.; Cerny, Anna; Asher, Damon R.; Kurt-Jones, Evelyn A.; Bronson, Roderick T.; Finberg, Robert W.



Temporal changes in stem cells in the circulation and myocardium of mice with Coxsackie virus B3-induced myocarditis  

Microsoft Academic Search

Our goal was to investigate temporal changes in stem cell in the circulation and myocardium of mice with Coxsackie virus B3-induced myocarditis. Groups of mice were administered Eagle's minimal essential medium or virus solution. The animals were further divided into six subgroups based on the following time points post-inoculation: 1, 3, 7, 14, 21, and 28 days. Ten animals were studied

Lan Zhao; Shuangjie Li; Junbo Ge; Aijun Sun; Yunzeng Zou; Shaoheng Zhang



Isoform-specific regulation and localization of the coxsackie and adenovirus receptor in human airway epithelia.  


Adenovirus is an important respiratory pathogen. Adenovirus fiber from most serotypes co-opts the Coxsackie-Adenovirus Receptor (CAR) to bind and enter cells. However, CAR is a cell adhesion molecule localized on the basolateral membrane of polarized epithelia. Separation from the lumen of the airways by tight junctions renders airway epithelia resistant to inhaled adenovirus infection. Although a role for CAR in viral spread and egress has been established, the mechanism of initial respiratory infection remains controversial. CAR exists in several protein isoforms including two transmembrane isoforms that differ only at the carboxy-terminus (CAR(Ex7) and CAR(Ex8)). We found low-level expression of the CAR(Ex8) isoform in well-differentiated human airway epithelia. Surprisingly, in contrast to CAR(Ex7), CAR(Ex8) localizes to the apical membrane of epithelia where it augments adenovirus infection. Interestingly, despite sharing a similar class of PDZ-binding domain with CAR(Ex7), CAR(Ex8) differentially interacts with PICK1, PSD-95, and MAGI-1b. MAGI-1b appears to stoichiometrically regulate the degradation of CAR(Ex8) providing a potential mechanism for the apical localization of CAR(Ex8) in airway epithelial. In summary, apical localization of CAR(Ex8) may be responsible for initiation of respiratory adenoviral infections and this localization appears to be regulated by interactions with PDZ-domain containing proteins. PMID:20361046

Excoffon, Katherine J D A; Gansemer, Nicholas D; Mobily, Matthew E; Karp, Philip H; Parekh, Kalpaj R; Zabner, Joseph



Outbreak of acute hemorrhagic conjunctivitis due to coxsackie A24 variant--Taiwan.  


Outbreaks of acute hemorrhagic conjunctivitis caused by enterovirus 70 and several serotypes of adenovirus have occurred in Taiwan since 1971. In 1980-1981, there was a pandemic of acute hemorrhagic conjunctivitis in southeast Asia caused by coxsackie A24 variant (CA24v); however, this virus did not affect Taiwan. In October 1985, CA24v was isolated for the first time from patients with acute hemorrhagic conjunctivitis in southern Taiwan. The following summer, a large epidemic of acute hemorrhagic conjunctivitis due to CA24v occurred. An epidemiologic investigation of patients seen at one ophthalmology clinic in Taipei City revealed that school-age children were the most likely group to introduce illness into households (p less than 0.001) and that males were more often household index cases than were females (p less than 0.01). Multiple case households tended to be more crowded (3.0 vs. 2.5 persons per bathroom; p less than 0.05) and had illness introduced by younger family members (median age of index case = 10 vs. 17 years; p less than 0.01). It is unknown whether this outbreak is an isolated occurrence or represents another resurgence of CA24v in the area. PMID:2833097

Chou, M Y; Malison, M D



Immunological and Biochemical Characterization of Coxsackie Virus A16 Viral Particles  

PubMed Central

Background Coxsackie virus A16 (CVA16) infections have become a serious public health problem in the Asia-Pacific region. It manifests most often in childhood exanthema, commonly known as hand-foot-and-mouth disease (HFMD). There are currently no vaccine or effective medical treatments available. Principal Finding In this study, we describe the production, purification and characterization of CVA16 virus produced from Vero cells grown on 5 g/L Cytodex 1 microcarrier beads in a five-liter serum-free bioreactor system. The viral titer was found to be >106 the tissue culture's infectious dose (TCID50) per mL within 7 days post-infection when a multiplicity of infection (MOI) of 10?5 was used for initial infection. Two CVA16 virus fractions were separated and detected when the harvested CVA16 viral concentrate was purified by a sucrose gradient zonal ultracentrifugation. The viral particles detected in the 24–28% sucrose fractions had low viral infectivity and RNA content. The viral particles obtained from 35–38% sucrose fractions were found to have high viral infectivity and RNA content, and composed of four viral proteins (VP1, VP2, VP3 and VP4), as shown by SDS-PAGE analyses. These two virus fractions were formalin-inactivated and only the infectious particle fraction was found to be capable of inducing CVA16-specific neutralizing antibody responses in both mouse and rabbit immunogenicity studies. But these antisera failed to neutralize enterovirus 71. In addition, rabbit antisera did not react with any peptides derived from CVA16 capsid proteins. Mouse antisera recognized a single linear immunodominant epitope of VP3 corresponding to residues 176–190. Conclusion These results provide important information for cell-based CVA16 vaccine development. To eliminate HFMD, a bivalent EV71/CVA16 vaccine formulation is necessary. PMID:23226233

Chong, Pele; Guo, Meng-Shin; Lin, Fion Hsiao-Yu; Hsiao, Kuang-Nan; Weng, Shu-Yang; Chou, Ai-Hsiang; Wang, Jen-Ren; Hsieh, Shih-Yang; Su, Ih-Jen; Liu, Chia-Chyi



Myocarditis: an expected health hazard associated with water resources contaminated with Coxsackie viruses type B.  


Enteroviruses, especially Coxsackie B viruses (CBVs), are responsible for approximately 50% of cases of viral myocarditis. In the present study, serum samples (160) were collected from acute myocarditis patients at different age groups and 104 samples of the same age groups as a control. Cholesterol, LDH, CPK, and GOT were measured for all serum samples (264). Also, to study the source of virus transmission, 72 water and 72 wastewater samples were collected from water and wastewater treatment plants at intakes and outlets. Water and wastewater samples were concentrated by filtration through Zeta-plus filter cartridges and reconcentrated by the PEG-6000 precipitation method. Serum, water, and wastewater samples were inoculated in BGM cells for three successive passages. RT-PCR with enterovirus primers was carried out directly for serum samples and for 1st and 3rd cell culture passages. The positive samples were used for neutralization assay using anti-CBV sera pool to determine the CBV followed by neutralization with separate antisera. The results showed that 50 (31.25%) serum samples from acute myocarditis patients and two (1.4%) samples from the controls were positive for enterovirus RT-PCR. For water and wastewater samples enteroviruses were present in 63.8% and 8.3% for intake and outlet of water treatment plants and, 66.6% and 47.2% for intake and outlet of wastewater treatment plants, respectively. The level of CBV serotypes was varied where CBV3 was dominant for all age groups of myocarditis patients and CBV2 and CBV5 were also detected while CBV2 was the main CBV in water samples and CBV2, 3 and 5 were detected in wastewater samples. The integration of cell culture-PCR reduces the time required for virus detection and enhances the sensitivity of the test. PMID:12909557

Ali, M A; Abdel-Dayem, T M K



Pathological Changes in Pregnant Mice Infected with Coxsackie B3 Virus as a Possible Cause of Retarded Foetal Development  

PubMed Central

Coxsackie B3 virus injected into mice on the eighth day of pregnancy resulted in foetal wastage and growth retardation. Although in apparent good health, the pregnant animals ate more food than the controls yet failed to increase in body weight as normal. This observation, together with the maternal autopsy findings of pancreatic acinar atrophy and hepatitis, suggests that the animals are subject to a manifestation of dietary deficiency attributable to an inability to break down and digest protein in their diet. It would seem that whilst the possibility of the virus exerting a direct effect on the foetuses cannot be ignored, the action of the virus in reducing the state of health of the pregnant mother is largely responsible for the foetal effects seen. ImagesFigs. 2-4Figs. 5-7Figs. 8-10 PMID:4857690

Lansdown, A. B. G.; Coid, C. R.



Cold Pressure Test Producing Coronary Spasm, Coronary Thrombosis and Myocardial Infarction in a Patient with IgM Antibodies against Coxsackie B Virus  

Microsoft Academic Search

Several lines of evidence have shown that viral infections are capable of causing coronary spasm and precipitating or mimicking clinical myocardial infarction. Here we report the case of a 41-year-old woman with recurrent angina who was admitted to our hospital because of ventricular tachycardia. Laboratory examination revealed positive IgM titers against Coxsackie B virus. Coronary angiography showed normal coronary arteries,

Werner Haberbosch; Norbert Roerich; Joerg Neuzner



Targeting Adenoviral Vectors by Using the Extracellular Domain of the Coxsackie-Adenovirus Receptor: Improved Potency via Trimerization  

PubMed Central

Adenovirus binds to mammalian cells via interaction of fiber with the coxsackie-adenovirus receptor (CAR). Redirecting adenoviral vectors to enter target cells via new receptors has the advantage of increasing the efficiency of gene delivery and reducing nonspecific transduction of untargeted tissues. In an attempt to reach this goal, we have produced bifunctional molecules with soluble CAR (sCAR), which is the extracellular domain of CAR fused to peptide-targeting ligands. Two peptide-targeting ligands have been evaluated: a cyclic RGD peptide (cRGD) and the receptor-binding domain of apolipoprotein E (ApoE). Human diploid fibroblasts (HDF) are poorly transduced by adenovirus due to a lack of CAR on the surface. Addition of the sCAR-cRGD or sCAR-ApoE targeting protein to adenovirus redirected binding to the appropriate receptor on HDF. However, a large excess of the monomeric protein was needed for maximal transduction, indicating a suboptimal interaction. To improve interaction of sCAR with the fiber knob, an isoleucine GCN4 trimerization domain was introduced, and trimerization was verified by cross-linking analysis. Trimerized sCAR proteins were significantly better at interacting with fiber and inhibiting binding to HeLa cells. Trimeric sCAR proteins containing cRGD and ApoE were more efficient at transducing HDF in vitro than the monomeric proteins. In addition, the trimerized sCAR protein without targeting ligands efficiently blocked liver gene transfer in normal C57BL/6 mice. However, addition of either ligand failed to retarget the liver in vivo. One explanation may be the large complex size, which serves to decrease the bioavailability of the trimeric sCAR-adenovirus complexes. In summary, we have demonstrated that trimerization of sCAR proteins can significantly improve the potency of this targeting approach in altering vector tropism in vitro and allow the efficient blocking of liver gene transfer in vivo. PMID:11799184

Kim, Jin; Smith,*, Theodore; Idamakanti, Neeraja; Mulgrew, Kathy; Kaloss, Michele; Kylefjord, Helen; Ryan, Patricia C.; Kaleko, Michael; Stevenson, Susan C.



Coxsackie- and adenovirus receptor (CAR) is expressed in lymphatic vessels in human skin and affects lymphatic endothelial cell function in vitro  

SciTech Connect

Lymphatic vessels play an important role in tissue fluid homeostasis, intestinal fat absorption and immunosurveillance. Furthermore, they are involved in pathologic conditions, such as tumor cell metastasis and chronic inflammation. In comparison to blood vessels, the molecular phenotype of lymphatic vessels is less well characterized. Performing comparative gene expression analysis we have recently found that coxsackie- and adenovirus receptor (CAR) is significantly more highly expressed in cultured human, skin-derived lymphatic endothelial cells (LECs), as compared to blood vascular endothelial cells. Here, we have confirmed these results at the protein level, using Western blot and FACS analysis. Immunofluorescence performed on human skin confirmed that CAR is expressed at detectable levels in lymphatic vessels, but not in blood vessels. To address the functional significance of CAR expression, we modulated CAR expression levels in cultured LECs in vitro by siRNA- and vector-based transfection approaches. Functional assays performed with the transfected cells revealed that CAR is involved in distinct cellular processes in LECs, such as cell adhesion, migration, tube formation and the control of vascular permeability. In contrast, no effect of CAR on LEC proliferation was observed. Overall, our data suggest that CAR stabilizes LEC-LEC interactions in the skin and may contribute to lymphatic vessel integrity.

Vigl, Benjamin; Zgraggen, Claudia; Rehman, Nadia; Banziger-Tobler, Nadia E.; Detmar, Michael [Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology, ETH Zurich, Wolfgang-Pauli Str. 10, CH-8093 Zurich (Switzerland); Halin, Cornelia [Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology, ETH Zurich, Wolfgang-Pauli Str. 10, CH-8093 Zurich (Switzerland)], E-mail:



The ?v?5 integrin of hematopoietic and nonhematopoietic cells is a transduction receptor of RGD-4C fiber-modified adenoviruses  

Microsoft Academic Search

Epithelial and endothelial cells expressing the primary Coxsackie virus B adenovirus (Ad) receptor (CAR) and integrin coreceptors are natural targets of human Ad infections. The fiber knob of species A, C, D, E and F Ad serotypes binds CAR by mimicking the CAR–homodimer interface, and the penton base containing arginine–glycine–aspartate (RGD) motifs binds with low affinity to ?v integrins inducing

H Nagel; S Maag; A Tassis; F O Nestlé; U F Greber; S Hemmi



The Murine CAR Homolog Is a Receptor for Coxsackie B Viruses and Adenoviruses  

Microsoft Academic Search

Complementary DNA clones encoding the murine homolog (mCAR) of the human coxsackievirus and adenovirus receptor (CAR) were isolated. Nonpermissive CHO cells transfected with mCAR cDNA became susceptible to infection by coxsackieviruses B3 and B4 and showed increased susceptibility to adenovirus- mediated gene transfer. These results indicate that the same receptor is responsible for virus interactions with both murine and human




[CANCER RESEARCH 63, 847853, February 15, 2003] Modulation of Coxsackie-Adenovirus Receptor Expression for Increased Adenoviral  

E-print Network

. Unfortunately, expression may often be low in advanced clinical cancers, including ovarian, colorectal, lung- pression by incubating ovarian cancer cells with various agents and then performing transgene expression in purified clinical ovarian cancer specimens. Possible clinical application was tested using i.p. administra

Hemminki, Akseli


First report of a Chinese strain of coxsackie B3 virus infection in a newborn in Germany in 2011: a case report  

PubMed Central

Introduction Enteroviruses commonly encounter babies and children and infections present in a wide variety of symptoms ranging from asymptomatic infection, benign illness, and aseptic meningitis, hand-foot-and-mouth disease to severe life-threatening disease. Some newborns develop severe disease in the first 2 weeks of life and long-term sequelae may occur among survivors. Case presentation We present a case report of a Caucasian newborn baby boy with severe encephalitis and systemic coxsackievirus B3 infection. The coincidence of maternal infection as well as previous mild respiratory illness in his sister suggests either prenatal or horizontal postnatal transmission. An electroencephalogram showed a severe pathologic pattern with theta-delta-rhythm and spike-wave complexes on both hemispheres. We also observed an unusual prolonged viremia for a period of 6 weeks. Due to the lack of specific antiviral treatment options, the supportive management included ventilation and medical treatment of seizures. Phylogenetic analysis revealed a genogroup D2 virus previously exclusively detected in China and now described in Europe for the first time. Conclusions Enteroviral infection is an important differential diagnosis in neonatal encephalitis. Prolonged viremia must be taken into account and might correlate with disease severity. The newly observed enterovirus genotype D2 is spreading from Asia to other continents. PMID:24885145



[Outbreak of acute enterovirus intestinal infection in Sakhalin region in August 2010].  


The investigation of cases of acute intestinal infections in the Sakhalin region of Russia in August, 2010 is described. Epidemiological and molecular biological studies were conducted. After initial PCR screening and determining the nucleotide sequences of the positive samples the following enteroviruses were found: Coxsackie A2 - 42 samples (45%), Coxsackie A4--31 sample (34%), Enterovirus 71--6 samples (6,5%), Coxsackievirus B5--6 samples (6,5%), Coxsackie B3--4 samples (4%) and Coxsackie B1--4 samples (4%). The phylogenetic analysis of sequences showed that the closest analogues for the nucleotide sequences of these genotypes were previously identified in Japan, Korea and China in 2000-2010. PMID:22642180

Demina, A V; Ternovo?, V A; Darizhapov, B B; Iakubich, T V; Sementsova, A O; Demina, O K; Protopopova, E V; Loktev, V B; Agafonov, A P; Netesov, S V



ORIGINAL ARTICLE Occurrence of adenovirus and other enteric viruses in  

E-print Network

ORIGINAL ARTICLE Occurrence of adenovirus and other enteric viruses in limited-contact freshwater Introduction Recreational outbreaks caused by adenoviruses, coxsackie- viruses, echoviruses and noroviruses (CSOs) and stormwater are sources of viruses which could significantly impact recreational water quality

Illinois at Chicago, University of


Integrin ?v?5 is a primary receptor for adenovirus in CAR-negative cells  

Microsoft Academic Search

BACKGROUND: Viruses bind to specific cellular receptors in order to infect their hosts. The specific receptors a virus uses are important factors in determining host range, cellular tropism, and pathogenesis. For adenovirus, the existing model of entry requires two receptor interactions. First, the viral fiber protein binds Coxsackie and Adenovirus Receptor (CAR), its primary cellular receptor, which docks the virus

Cynthia Lyle; Frank McCormick



379. Identification of Structural Aspects of Adenovirus Interactions with Host Factors In Vivo Using Tandem Mass Spectrometry  

Microsoft Academic Search

Intensive studies of adenovirus infectivity and bio-distribution in animal models demonstrated that liver tissue is responsible for clearance of the bulk of intravenously applied vector. We recently demonstrated that adenovirus binding to blood factors, specifically coagulation factor IX and C4 binding protein, leads to high level hepatocyte transduction and virus trapping by Kupffer cells. Apparently, virus interaction with the coxsackie-

Oleksandr Kalyuzhniy; Catalin Doneanu; Martin Sadilek; Dmitry M. Shayakhmetov



National Laboratory Poornima Upadhya  

E-print Network

of Energy. Competitive Advantage The technology can be used to engineer viral tropism. In additionBrookhaven National Laboratory Poornima Upadhya Office of Intellectual Property and Sponsored address: Human Coxsackie and Adenovirus Receptor - CAR Brookhaven National Laboratory


Benzophenone C-glucosides and gallotannins from mango tree stem bark with broad-spectrum anti-viral activity.  


The high mutation rate of RNA viruses has resulted in limitation of vaccine effectiveness and increased emergence of drug-resistant viruses. New effective antivirals are therefore needed to control of the highly mutative RNA viruses. The n-butanol fraction of the stem bark of Mangifera indica exhibited inhibitory activity against influenza neuraminidase (NA) and coxsackie virus 3C protease. Bioassay guided phytochemical study of M. indica stem bark afforded two new compounds including one benzophenone C-glycoside (4) and one xanthone dimer (7), together with eleven known compounds. The structures of these isolated compounds were elucidated on the basis of spectroscopic evidences and correlated with known compounds. Anti-influenza and anti-coxsackie virus activities were evaluated by determining the inhibition of anti-influenza neuraminidase (NA) from pandemic A/RI/5+/1957 H2N2 influenza A virus and inhibition of coxsackie B3 virus 3C protease, respectively. The highest anti-influenza activity was observed for compounds 8 and 9 with IC50 values of 11.9 and 9.2?M, respectively. Compounds 8 and 9 were even more potent against coxsackie B3 virus 3C protease, with IC50 values of 1.1 and 2.0?M, respectively. Compounds 8 and 9 showed weak cytotoxic effect against human hepatocellular carcinoma and human epithelial carcinoma cell lines through MTT assay. PMID:24613627

Abdel-Mageed, Wael M; Bayoumi, Soad A H; Chen, Caixia; Vavricka, Christopher J; Li, Li; Malik, Ajamaluddin; Dai, Huanqin; Song, Fuhang; Wang, Luoqiang; Zhang, Jingyu; Gao, George F; Lv, Yali; Liu, Lihong; Liu, Xueting; Sayed, Hanaa M; Zhang, Lixin



Action of commonly used disinfectants against enteroviruses.  


The virucidal effect of some of the most commonly used hospital disinfectants against Coxsackie B4, Echovirus 11, Poliovirus type 1 and Rotavirus have been evaluated. It was found that 'Chloros', 'Totacide 28' and methylated spirits were completely virucidal to all the viruses under study. 'Stericol' and 'Lysol' had a limited effect while 'Hibiscrub' and 'Savlon' had no effect at all. PMID:6195231

Narang, H K; Codd, A A



TRENDS IN MOLECULAR MEDICINE Adenoviruses for treatment of cancer  

E-print Network

that the primary receptor, the coxsackie-adenovirus receptor (CAR) expression in tumors may be highly variable to modify adenovirus tropism in order to circumvent CAR deficiency, including retargeting complexes or genetic capsid modifications. To further improve tumor penetration and local amplification of the anti-tumor

Hemminki, Akseli


Detection and Molecular Typing of Enteroviruses in Water Sources  

Microsoft Academic Search

In view of the major health impact of enteroviruses, sensitive and reliable techniques for the detection and identification of this large group of closely related viruses are essential. To date enteroviruses have generally been isolated by cell culture propagation and typed by neutralization tests using specific antisera. Inoculation of newborn mice remains the most common procedure to distinguish between Coxsackie

JC Vivier; CG Clay; WOK Grabow



CAR mediates efficient tumor engraftment of mesenchymal type lung cancer cells  

Microsoft Academic Search

The coxsackie-adenovirus receptor (CAR) is a developmentally regulated intercellular adhesion molecule that was previously observed to be required for efficient tumor formation. To confirm that observation, we compared the tumorigenicity of clonally derived test and control cell subsets that were genetically modified for CAR. Silencing CAR in lung cancer cells with high constitutive expression reduced engraftment efficiency. Conversely, overexpressing CAR

Mysore S Veena; Min Qin; Åsa Andersson; Sherven Sharma; Raj K Batra



Genome stability of adenovirus types 3 and 7 during a simultaneous outbreak in Greater Manchester, UK.  


A total of 96 isolates of species B adenovirus collected in Greater Manchester, UK and typed previously by serum neutralization were analyzed in five genome regions. Of these, 62 isolates were HAdV-B3 and HAdV-B7 collected during a simultaneous 15 months outbreak. The rest of the isolates were HAdV-B types 3 and 7 and other species B adenovirus types collected in different years following the outbreak. The phylogenetic analysis results of all the isolates in the structural regions hexon L2, penton, and fiber knob were found to be consistent and no mismatches were observed. Most of the isolates in the DNA polymerase and E1A regions had the same clustering patterns as the structural regions. However, one HAdV-B7 and one HAdV-B11 isolate changed their clustering patterns in the DNA polymerase region. In addition, HAdV-B16 isolates changed their clustering patterns in both DNA polymerase and E1A regions. The changes of the clustering patterns of some isolates is more likely related to natural variations rather than recombination which indicate that species B adenovirus genome is stable even when different types are circulating in a limited geographical area simultaneously. J. Med. Virol. 87: 117-124, 2015. © 2014 Wiley Periodicals, Inc. PMID:24801279

Alkhalaf, Moustafa Alissa; Guiver, Malcolm; Cooper, Robert J



High-throughput screening of viral entry inhibitors using pseudotyped virus.  


Virus entry into a host cell is an attractive target for therapy because propagation of virus can be blocked at an early stage, minimizing chances for the virus to acquire drug resistance. Anti-infective drug discovery for BSL-4 viruses like Ebola or Lassa hemorrhagic fever virus presents challenges due to the requirement for a BSL-4 laboratory containment facility. Pseudotyped viruses provide a surrogate model in which the native envelope glycoprotein of a BSL-2 level virus (e.g., vesicular stomatitis virus) is replaced with envelope glycoprotein of a foreign BSL-4 virus (e.g., Ebola virus). Because the envelope glycoprotein determines interaction of virus with its cellular receptors, pseudotyped viruses can mimic the viral entry process of the original virus. Moreover, they are competent for only a single cycle of infection, and therefore can be used in BSL-2 facilities. Pseudotyped viruses have been used in high-throughput screening of entry inhibitors for a number of BSL-4 level viruses. This unit includes protocols for preparing pseudotyped viruses using lentiviral vectors and use of pseudotyped viruses for high-throughput screening of viral entry inhibitors. PMID:21935898

Basu, Arnab; Mills, Debra M; Bowlin, Terry L



THE YALE JOURNAL OF BIOLOGY AND MEDICINE 62 (1989), 67-75 Bibliography  

E-print Network

4. Hsiung GD, Stafseth HJ: Canine distemper. II. The value of certain therapeutic agents against the bacterial infections in dogs affected with distemper. Cornell Vet 42:232-237, 1952 5. Hsiung GD, Haley LD: A comparative study in the isolation of Mycobacterium tuberculosis on artificial media and embryonated eggs. Am Rev Tuber 70:912-915, 1954 6. Hsiung GD, Haley LD: A modification on virulence of Mycobacterium tuberculosis by chick embryo adaptation. Am Rev Tuber 74:249-257, 1956 7. Hsiung GD, Melnick JL: Plaque formation with poliomyelitis, coxsackie and orphan (Echo) viruses in bottle cultures of monkey epithelial cells. Virology 1:533-535, 1955 8. Hsiung GD, Melnick JL: Morphologic characteristics of plaques produced on monkey kidney monolayer cultures by enteric viruses (poliomyelitis, coxsackie, and ECHO groups). J Immunol

G. D. Edith Hsiung, Ph.D.; Mich Agri; Exp Sta


Collection and Testing of Respiratory Samples

QIAGEN ResPlex II Advanced Panel; Influenza A; Influenza B; Respiratory Syncytial Virus Infections; Infection Due to Human Parainfluenza Virus 1; Parainfluenza Type 2; Parainfluenza Type 3; Parainfluenza Type 4; Human Metapneumovirus A/B; Rhinovirus; Coxsackie Virus/Echovirus; Adenovirus Types B/C/E; Coronavirus Subtypes 229E; Coronavirus Subtype NL63; Coronavirus Subtype OC43; Coronavirus Subtype HKU1; Human Bocavirus; Artus Influenza A/B RT-PCR Test; Influenza A, Influenza B,



Adenovirus triggers macropinocytosis and endosomal leakage together with its clathrin-mediated uptake  

Microsoft Academic Search

denovirus type 2 (Ad2) binds the coxsackie B virus Ad receptor and is endocytosed upon activation of thev integrin coreceptors. Here, we demonstrate that expression of dominant negative clathrin hub, eps15, or K44A-dynamin (dyn) inhibited Ad2 uptake into epithe- lial cells, indicating clathrin-dependent viral endocytosis. Surprisingly, Ad strongly stimulated the endocytic uptake of fluid phase tracers, coincident with virus internalization

Oliver Meier; Karin Boucke; Silvija Vig Hammer; Stephan Keller; Robert P. Stidwill; Silvio Hemmi; Urs F. Greber



Psoriasis Herpeticum due to Varicella Zoster Virus: A Kaposi's Varicelliform Eruption in Erythrodermic Psoriasis  

PubMed Central

Kaposi's varicelliform eruption (KVE) or eczema herpeticum is characterized by disseminated papulovesicular eruption caused by a number of viruses like Herpes simplex virus I and II, Coxsackie virus, and Vaccinia and Small pox viruses in patients with pre-existing skin disease. The occurrence of KVE with psoriasis has been reported recently as a new entity psoriasis herpeticum. The rare causation of psoriasis herpeticum due to Varicella zoster virus in a patient with underlying psoriasis is being reported for the first time. PMID:22707775

Garg, Geeta; Thami, Gurvinder P



Antimicrobial and cytotoxic activities of turbinariaconoides (j.agardh) kuetz.  


Brown alga, Turbinariaconoideswas successively extracted with n-hexane, cyclohexane, methanol and ethanol:water (1:1). The extracts were evaluated for antibacterial and antifungal activities by disc diffusion method. Minimal inhibitory concentration was determined for active extracts by broth dilution method. The antiviral activity and cytotoxicity of the extracts were tested in human embryonic lung (HEL) cells (herpes simplex virus-1, herpes simplex virus-2, vaccinia virus, vesicular stomatitis virus and herpes simplex virus-1 TK- KOS ACVr), human epithelial (HeLa) cells (vesicular stomatitis virus and coxsackie virus B4) and Vero cells (parainfluenza-3 virus, reovirus-1, sindbis virus coxsackie virus B4 and puntatoro virus). The results revealed that extracts exhibited cytotoxicity ranged from 20 to >100 ?g/mL. Moderate activity was demonstrated by n-hexane and cyclohexane extracts against viruses, whereas methanol and ethanol:water (1:1) extracts were not active. Ethanol:water (1:1) presented neither antibacterial nor antifungal activity against tested organisms. Cyclohexane extract possessed a broad array of antibacterial activity and exhibited remarkable antifungal property. It is noteworthy that minimal inhibitory concentration of cyclohexane extract against Aspergillusnigeris comparable with that of clotrimazole. This potentiality demonstrates that it could be used to treat bacterial and fungal infections. PMID:24381606

Kumar Shanmugam, Sadish; Kumar, Yatendra; Sardaryar, Khan Mohammad; Gupta, Vivek; De Clercq, Erik



Seroprevalence of Human Enterovirus 71 and Coxsackievirus A16 in Guangdong, China, in Pre- and Post-2010 HFMD Epidemic Period  

PubMed Central

Background Human Enterovirus 71 and Coxsackie A16 have caused many outbreaks in the last decade in mainland China, resulting in thousands of fatal cases. Seroepidemiology which provides important information to document population immunity is rare in China. Methodology/Principal Findings A cross sectional study of Enterovirus 71 (EV71) and Coxsackie A16 (CA16) seroprevalence was carried out in Guangdong, China, pre- and post- the 2010 hand, foot and mouth disease (HFMD) epidemic period. The levels of EV71 and CA16 specific antibodies were evaluated by a microneutralization test and the geometric mean titer (GMT) was calculated and compared. Our results indicated frequent infection by EV71 and CA16 in Guangdong before the 2010 epidemic. Only EV71 neutralizing antibody but not CA16 seroprevalence was significantly increased after the 2010 HFMD epidemic. Children less than 3 years old especially those aged 2 years showed the lowest positive rates for EV71 and CA16 NA before epidemic and the most significantly increased EV71 seroprevalence after epidemic. CA16 GMT values declined after the 2010 epidemic. Conclusions These results indicate EV71 was the major pathogen of HFMD in Guangdong during the 2010 epidemic. The infection occurs largely in children less than 3 years, who should have first priority to receive an EV71 vaccine. PMID:24324604

Su, Juan; Lu, Jing; Ke, Changwen; Zeng, Hanri; Guan, Dawei; Ma, Cong; Zhang, Wanly; Xiao, Hong; Li, Hui; Lin, Jinyan; Zhang, Yonghui



Chest wall myositis in a patient with acute coronary syndrome.  


We describe a case of a 42-year-old man who presented to the emergency department with severe left-sided chest pain and chest tenderness of 1-day duration. The pain was episodic and was aggravated by any chest wall movement. His initial blood tests and ECG were suggestive of acute coronary syndrome (ACS). However, his pattern of pain, lack of response to opiates, raised creatine kinase and signs of pleurisy on chest radiograph raised a suspicion of an alternative diagnosis. The patient showed a dramatic response in pain relief to non-steroidal anti-inflammatory medication. He was suspected to have chest wall myositis with pleural involvement in the form of pleurodynia. His serology test was positive for coxsackie virus antibodies. We will discuss in this case report the pathognomonic features, diagnosis and treatment of a rare infectious condition known as Bornholm disease. PMID:25312897

Hussein, Laila; Al-Rawi, Harith



Detection of Viral Pathogens by Reverse Transcriptase PCR and of Microbial Indicators by Standard Methods in the Canals of the Florida Keys  

PubMed Central

In order to assess the microbial water quality in canal waters throughout the Florida Keys, a survey was conducted to determine the concentration of microbial fecal indicators and the presence of human pathogenic microorganisms. A total of 19 sites, including 17 canal sites and 2 nearshore water sites, were assayed for total coliforms, fecal coliforms, Escherichia coli, Clostridium perfringens, enterococci, coliphages, F-specific (F+) RNA coliphages, Giardia lamblia, Cryptosporidium parvum, and human enteric viruses (polioviruses, coxsackie A and B viruses, echoviruses, hepatitis A viruses, Norwalk viruses, and small round-structured viruses). Numbers of coliforms ranged from <1 to 1,410, E. coli organisms from <1 to 130, Clostridium spp. from <1 to 520, and enterococci from <1 to 800 CFU/100 ml of sample. Two sites were positive for coliphages, but no F+ phages were identified. The sites were ranked according to microbial water quality and compared to various water quality standards and guidelines. Seventy-nine percent of the sites were positive for the presence of enteroviruses by reverse transcriptase PCR (polioviruses, coxsackie A and B viruses, and echoviruses). Sixty-three percent of the sites were positive for the presence of hepatitis A viruses. Ten percent of the sites were positive for the presence of Norwalk viruses. Ninety-five percent of the sites were positive for at least one of the virus groups. These results indicate that the canals and nearshore waters throughout the Florida Keys are being impacted by human fecal material carrying human enteric viruses through current wastewater treatment strategies such as septic tanks. Exposure to canal waters through recreation and work may be contributing to human health risks. PMID:10473424

Griffin, Dale W.; Gibson, Charles J.; Lipp, Erin K.; Riley, Kelley; Paul, John H.; Rose, Joan B.



Structural Basis for Antiviral Inhibition of the Main Protease, 3C, from Human Enterovirus 93 ?  

PubMed Central

Members of the Enterovirus genus of the Picornaviridae family are abundant, with common human pathogens that belong to the rhinovirus (HRV) and enterovirus (EV) species, including diverse echo-, coxsackie- and polioviruses. They cause a wide spectrum of clinical manifestations ranging from asymptomatic to severe diseases with neurological and/or cardiac manifestations. Pandemic outbreaks of EVs may be accompanied by meningitis and/or paralysis and can be fatal. However, no effective prophylaxis or antiviral treatment against most EVs is available. The EV RNA genome directs the synthesis of a single polyprotein that is autocatalytically processed into mature proteins at Gln?Gly cleavage sites by the 3C protease (3Cpro), which has narrow, conserved substrate specificity. These cleavages are essential for virus replication, making 3Cpro an excellent target for antivirus drug development. In this study, we report the first determination of the crystal structure of 3Cpro from an enterovirus B, EV-93, a recently identified pathogen, alone and in complex with the anti-HRV molecules compound 1 (AG7404) and rupintrivir (AG7088) at resolutions of 1.9, 1.3, and 1.5 Å, respectively. The EV-93 3Cpro adopts a chymotrypsin-like fold with a canonically configured oxyanion hole and a substrate binding pocket similar to that of rhino-, coxsackie- and poliovirus 3C proteases. We show that compound 1 and rupintrivir are both active against EV-93 in infected cells and inhibit the proteolytic activity of EV-93 3Cpro in vitro. These results provide a framework for further structure-guided optimization of the tested compounds to produce antiviral drugs against a broad range of EV species. PMID:21835784

Costenaro, Lionel; Kaczmarska, Zuzanna; Arnan, Carme; Janowski, Robert; Coutard, Bruno; Sola, Maria; Gorbalenya, Alexander E.; Norder, Helene; Canard, Bruno; Coll, Miquel



Synthesis, antiviral and cytotoxic activity of 6-bromo-2,3-disubstituted-4(3H)-quinazolinones.  


In the present study, a series of 6-bromo-2,3-disubstitued-4(3H)-quinazolinones was synthesized by condensation of 6-bromo-2-substituted-benzoxazin-4-one with trimethoprim, pyrimethamine and lamotrigine. The chemical structures of the synthesized compounds were confirmed by means of IR, (1)H-NMR and mass spectral and elemental analysis. The antiviral activity and cytotoxicity of the compounds were tested in E(6)SM (Herpes simplex-1 KOS, Herpes simplex-1 TK-KOS ACV, Herpes simplex-2 G, Vaccinia virus, Vesicular stomatitis virus, Parainfluenza-3 virus, Reovirus-1, Sindbis virus, Coxsackie virus B4 and Punta Toro virus) and HeLa cell culture (Vesicular stomatitis virus, Coxsackie virus B4 and Respiratory syncyticla virus). Investigation of anti-HIV activity was done against replication of HIV-1 (HTLV-III B LAI) in MT-4 cells. 6-Bromo-2-phenyl-3-[(4-amino-5-(4-chlorophenyl)-6-ethylpyrimidin-2-yl]-4(3H)-quinazolinone (4) exhibited the most potent antiviral activity with a MIC of 1.92 microg/ml against vaccinia virus in E(6)SM cell culture. The other compounds did not exhibit antiviral activity nor afford significant cytoprotection to the E(6)SM and HeLa cell culture when challenged with the viruses. The study implies that 4 may possess activity against Pox viruses including variola. In the anti-HIV study, 6-bromo-2-methyl-3-[(4-amino-5-(4-chlorophenyl)-6-ethylpyrimidin-2-yl]-4(3H)-quinazolinone (3) and 6-bromo-2-phenyl-3-[(4-amino-5-(4-chlorophenyl)-6-ethylpyrimidin-2-yl]-4(3H)-quinazolinone (4) exhibited the least cytotoxic concentration (0.424, 0.461 microg/ml) which is an index of the infective viability of mock infected MT-4 cells with HIV-1. None of the compounds exhibited significant anti-HIV activity. PMID:12951471

Dinakaran, Murugesan; Selvam, Periyaswamy; DeClercq, Erik; Sridhar, Seshaiah Krishnan



Anti-viral and anti-bacterial activities of an extract of blackcurrants (Ribes nigrum L.).  


The inhibitory effects of an extract of the blackcurrant (Ribes nigrum L.) against pathogens associated with oral, nasopharyngeal and upper respiratory infectious diseases; namely respiratory syncytial virus (RSV), influenza virus A and B (IFV-A and IFV-B), adenovirus (AdV), herpes simplex virus type 1, Haemophilus influenzae type B, Streptococcus pneumoniae and Streptococcus mutans, were investigated. Less than 1% concentration of extract of blackcurrant inhibited replication of RSV, IFV-A and -B and HSV-1 by over 50% and a 10% extract inhibited adsorption of these viruses onto the cell surface by over 95%. The effects on AdV were much less pronounced; the half minimal inhibitory concentration of AdV replication was 2.54 ± 0.26, and a 10% concentration of the extract inhibited AdV adsorption on the cell surface by 72.9 ± 3.4%. The antibacterial activities of the blackcurrant were evaluated based on its efficacy as a disinfectant. A 10% extract disinfected 99.8% of H. Influenzae type B and 78.9% of S. pneumoniae in 10 min, but had no demonstrable effect against S. mutans. The blackcurrant extract still showed antiviral and antibacterial activities after the pH had been made neutral with sodium hydroxide, suggesting that these activities are not the result of acidic reactions or of components precipitated at a neutral pH. These findings demonstrate the potential of blackcurrant extract as a functional food for oral care. PMID:22985050

Ikuta, Kazufumi; Hashimoto, Koichi; Kaneko, Hisatoshi; Mori, Shuichi; Ohashi, Kazutaka; Suzutani, Tatsuo



Adenovirus-pulsed dendritic cells stimulate human virus-specific T-cell responses in vitro.  

PubMed Central

Adenovirus infections cause significant morbidity and mortality in immunocompromised patients, yet little is known about the immune response to adenovirus infections. We established a system for the generation of a cytotoxic immune response to adenovirus in vitro. Cytotoxic T cells (CTLs) were derived from normal donors by using peripheral blood dendritic cells as antigen-presenting cells. The CTLs were found to contain a mixture of effector cells that recognized virus peptides in the context of both class I and class II antigens. Endogenous viral gene expression was not required to sensitize cells to lysis by adenovirus-specific CTLs. CTLs raised against subgroup C adenovirus type 5 can lyse cells infected with subgroup B adenovirus type 11, indicating that viruses of different subgroups have epitopes in common. This system holds promise for defining the human immune response to adenovirus, including characterization of the viral protein(s) against which the response is generated, and the identity of the effector cells. Such studies are in progress. PMID:8794310

Smith, C A; Woodruff, L S; Kitchingman, G R; Rooney, C M



Oxidative stress in coronary artery disease: epigenetic perspective.  


The association between oxidative stress and coronary artery disease (CAD) is well documented. However, the role of epigenetic factors contributing to oxidative stress is relatively unexplored. In this study, we aimed to explore the impact of DNA methylation profile in BCL2/E1B adenovirus interacting protein 3 (BNIP3), extracellular superoxide dismutase (EC-SOD) and glutathione-S-transferase P1 (GSTP1) on the oxidative stress in CAD. Further, the contribution of folate pathway genetic polymorphisms in regulating epigenome was elucidated. The expression of BNIP3, EC-SOD, and GSTP1 were studied by using Maxima@SYBR-green based real-time qPCR approach in peripheral blood samples. Combined bisulfite restriction analysis and methylation-specific PCR were used to study promoter CpG island methylation. Further, the effect of homocysteine on BNIP3 gene expression was studied in human aortic endothelial cells in vitro. CAD cases exhibited upregulation of BNIP3, downregulation of EC-SOD and GSTP1. Hypomethylation of BNIP3 and hypermethylation of EC-SOD were observed in CAD cases. The expression of BNIP3 was positively correlated with homocysteine, MDA, protein carbonyls, and methylene tetrahydrofolate reductase C677T, while showing inverse association with cytosolic serine hydroxymethyl transferase C1420T. The expressions of EC-SOD and GSTP1 showed positive association with thymidylate synthase (TYMS) 2R3R, while inverse association with MDA, protein carbonyls, and methionine synthase reductase (MTRR) A66G. In vitro analysis showed homocysteine-dependent upregulation of BNIP3. The results of this study suggest that the aberrations in one-carbon metabolism appear to induce altered gene expression of EC-SOD, GSTP1, and BNIP3, and thus contribute to the increased oxidative stress and increased susceptibility to CAD. PMID:23160801

Lakshmi, Sana Venkata Vijaya; Naushad, Shaik Mohammad; Reddy, Cheruku Apoorva; Saumya, Kankanala; Rao, Damera Seshagiri; Kotamraju, Srigiridhar; Kutala, Vijay Kumar



Vascular endothelial growth factor signalling in endothelial cell survival: A role for NF{kappa}B  

SciTech Connect

Angiogenesis is the development of blood capillaries from pre-existing vessels. Vascular endothelial growth factor (VEGF) is a key regulator of vessel growth and regression, and acts as an endothelial survival factor by protecting endothelial cells from apoptosis. Many genes involved in cell proliferation and apoptosis are regulated by the nuclear factor kappa B (NF{kappa}B) transcription factor family. This study aimed to address the hypothesis that VEGF-mediated survival effects on endothelium involve NF{kappa}B. Using an NF{kappa}B-luciferase reporter adenovirus, we observed activation of NF{kappa}B following VEGF treatment of human umbilical vein endothelial cells. This was confirmed using electrophoretic mobility shift assay and found to involve nuclear translocation of NF{kappa}B sub-unit p65. However, NF{kappa}B activation occurred without degradation of inhibitory I{kappa}B proteins (I{kappa}B{alpha}, I{kappa}B{beta}, and I{kappa}B{epsilon}). Instead, tyrosine phosphorylation of I{kappa}B{alpha} was observed following VEGF treatment, suggesting NF{kappa}B activation was mediated by degradation-independent dissociation of I{kappa}B{alpha} from NF{kappa}B. Adenovirus-mediated over-expression of either native I{kappa}B{alpha}, or of I{kappa}B{alpha} in which tyrosine residue 42 was mutated to phenylalanine, inhibited induction of NF{kappa}B-dependent luciferase activity in response to VEGF. Furthermore, VEGF-induced upregulation of mRNA for the anti-apoptotic protein Bcl-2 and cell survival following serum withdrawal was reduced following I{kappa}B{alpha} over-expression. This study highlights that different molecular mechanisms of NF{kappa}B activation may be involved downstream of stimuli which activate the endothelial lining of blood vessels.

Grosjean, Jennifer [Kennedy Institute of Rheumatology, Faculty of Medicine, Imperial College, London (United Kingdom)]. E-mail:; Kiriakidis, Serafim [Kennedy Institute of Rheumatology, Faculty of Medicine, Imperial College, London (United Kingdom); Reilly, Kerri [Kennedy Institute of Rheumatology, Faculty of Medicine, Imperial College, London (United Kingdom); Feldmann, Marc [Kennedy Institute of Rheumatology, Faculty of Medicine, Imperial College, London (United Kingdom); Paleolog, Ewa [Kennedy Institute of Rheumatology, Faculty of Medicine, Imperial College, London (United Kingdom)



Homomultimerization of the Coxsackievirus 2B Protein in Living Cells Visualized by Fluorescence Resonance Energy Transfer Microscopy  

PubMed Central

The 2B protein of enteroviruses is the viral membrane-active protein that is responsible for the modifications in host cell membrane permeability that take place in enterovirus-infected cells. The 2B protein shows structural similarities to the group of lytic polypeptides, polypeptides that permeate membranes either by forming multimeric membrane-integral pores or, alternatively, by lying parallel to the lipid bilayer and disturbing the curvature and symmetry of the membrane. Our aim is to gain more insight into the molecular architecture of the 2B protein in vivo. In this study, the possible existence of multimers of the coxsackie B3 virus 2B protein in single living cells was explored by fluorescence resonance energy transfer (FRET) microscopy. FRET between fusion proteins 2B-ECFP and 2B-EYFP (enhanced cyan and yellow fluorescent variants of green fluorescent protein) was monitored by using spectral imaging microscopy (SPIM) and fluorescence lifetime imaging microscopy (FLIM). Both techniques revealed the occurrence of intermolecular FRET between 2B-ECFP and 2B-EYFP, providing evidence for the formation of protein 2B homomultimers. Putative models for the mode of action of the membrane-active 2B protein and the formation of membrane-integral pores by 2B multimers are discussed. PMID:12186926

van Kuppeveld, Frank J. M.; Melchers, Willem J. G.; Willems, Peter H. G. M.; Gadella, Jr., Theodorus W. J.



Solution Structure of the 2A Protease from a Common Cold Agent, Human Rhinovirus C2, Strain W12  

PubMed Central

Human rhinovirus strains differ greatly in their virulence, and this has been correlated with the differing substrate specificity of the respective 2A protease (2Apro). Rhinoviruses use their 2Apro to cleave a spectrum of cellular proteins important to virus replication and anti-host activities. These enzymes share a chymotrypsin-like fold stabilized by a tetra-coordinated zinc ion. The catalytic triad consists of conserved Cys (C105), His (H34), and Asp (D18) residues. We used a semi-automated NMR protocol developed at NMRFAM to determine the solution structure of 2Apro (C105A variant) from an isolate of the clinically important rhinovirus C species (RV-C). The backbone of C2 2Apro superimposed closely (1.41–1.81 Å rmsd) with those of orthologs from RV-A2, coxsackie B4 (CB4), and enterovirus 71 (EV71) having sequence identities between 40% and 60%. Comparison of the structures suggest that the differential functional properties of C2 2Apro stem from its unique surface charge, high proportion of surface aromatics, and sequence surrounding the di-tyrosine flap. PMID:24937088

Lee, Woonghee; Watters, Kelly E.; Troupis, Andrew T.; Reinen, Nichole M.; Suchy, Fabian P.; Moyer, Kylie L.; Frederick, Ronnie O.; Tonelli, Marco; Aceti, David J.; Palmenberg, Ann C.; Markley, John L.



Treatment of Cancer Patients With a Serotype 5/3 Chimeric Oncolytic Adenovirus Expressing GMCSF  

PubMed Central

Augmenting antitumor immunity is a promising way to enhance the potency of oncolytic adenoviral therapy. Granulocyte–macrophage colony–stimulating factor (GMCSF) can mediate antitumor effects by recruiting natural killer cells and by induction of tumor-specific CD8+ cytotoxic T-lymphocytes. Serotype 5 adenoviruses (Ad5) are commonly used in cancer gene therapy. However, expression of the coxsackie-adenovirus receptor is variable in many advanced tumors and preclinical data have demonstrated an advantage for replacing the Ad5 knob with the Ad3 knob. Here, a 5/3 capsid chimeric and p16-Rb pathway selective oncolytic adenovirus coding for GMCSF was engineered and tested preclinically. A total of 21 patients with advanced solid tumors refractory to standard therapies were then treated intratumorally and intravenously with Ad5/3-D24-GMCSF, which was combined with low-dose metronomic cyclophosphamide to reduce regulatory T cells. No severe adverse events occurred. Analysis of pretreatment samples of malignant pleural effusion and ascites confirmed the efficacy of Ad5/3-D24-GMCSF in transduction and cell killing. Evidence of biological activity of the virus was seen in 13/21 patients and 8/12 showed objective clinical benefit as evaluated by radiology with Response Evaluation Criteria In Solid Tumors (RECIST) criteria. Antiadenoviral and antitumoral immune responses were elicited after treatment. Thus, Ad5/3-D24-GMCSF seems safe in treating cancer patients and promising signs of efficacy were seen. PMID:20664527

Koski, Anniina; Kangasniemi, Lotta; Escutenaire, Sophie; Pesonen, Sari; Cerullo, Vincenzo; Diaconu, Iulia; Nokisalmi, Petri; Raki, Mari; Rajecki, Maria; Guse, Kilian; Ranki, Tuuli; Oksanen, Minna; Holm, Sirkka-Liisa; Haavisto, Elina; Karioja-Kallio, Aila; Laasonen, Leena; Partanen, Kaarina; Ugolini, Matteo; Helminen, Andreas; Karli, Eerika; Hannuksela, Paivi; Pesonen, Saila; Joensuu, Timo; Kanerva, Anna; Hemminki, Akseli



The novel complement inhibitor human CUB and Sushi multiple domains 1 (CSMD1) protein promotes factor I-mediated degradation of C4b and C3b and inhibits the membrane attack complex assembly.  


CUB and Sushi multiple domains 1 (CSMD1) is a transmembrane protein containing 15 consecutive complement control protein (CCP) domains, which are characteristic for complement inhibitors. We expressed a membrane-bound fragment of human CSMD1 composed of the 15 C-terminal CCP domains and demonstrated that it inhibits deposition of C3b by the classical pathway on the surface of Chinese hamster ovary cells by 70% at 6% serum and of C9 (component of membrane attack complex) by 90% at 1.25% serum. Furthermore, this fragment of CSMD1 served as a cofactor to factor I-mediated degradation of C3b. In all functional assays performed, well-characterized complement inhibitors were used as positive controls, whereas Coxsackie adenovirus receptor, a protein with no effect on complement, was a negative control. Moreover, attenuation of expression in human T47 breast cancer cells that express endogenous CSMD1 significantly increased C3b deposition on these cells by 45% at 8% serum compared with that for the controls. Furthermore, by expressing a soluble 17-21 CCP fragment of CSMD1, we found that CSMD1 inhibits complement by promoting factor I-mediated C4b/C3b degradation and inhibition of MAC assembly at the level of C7. Our results revealed a novel complement inhibitor for the classical and lectin pathways. PMID:23964079

Escudero-Esparza, Astrid; Kalchishkova, Nikolina; Kurbasic, Emila; Jiang, Wen G; Blom, Anna M



Exploring the dynamics of four RNA-dependent RNA polymerases by a coarse-grained model.  


In this article, we present a hybrid ENM/MARTINI coarse-grained model and examine the impact of reduced chemical detail on both static and dynamic properties by comparing against explicit atomistic simulations. This methodology complements the advanced molecular characterization and dynamics of proteins for medical and bioengineering applications by developing a fundamental understanding of how the motion and molecular characteristics of proteins, viruses, their precursors, and their interactions with the environment govern their behavior in different populations. As an example, we explore the dynamics of RNA-dependent RNA polymerases (RdRPs) from the following viruses: poliovirus, Coxsackie virus B3, human rhinovirus 16, and foot-and-mouth-disease virus. The hybrid coarse-grained model allows the microsecond time scales of interest for biological functions to be explored. Additionally, the ENM/MARTINI model captures the main features obtained from atomistic MD simulations for each of the RdRPs studied herein, including the higher flexibility of the pinky finger and thumb regions, as well as collective motions that might contribute significantly to the conformational transition between the open and closed states. PMID:23157550

Shen, Hujun; Moustafa, Ibrahim M; Cameron, Craig E; Colina, Coray M



Idiopathic Infantile Arterial Calcification: A Possible Cause of Refractory Cardiopulmonary Failure in Infancy  

PubMed Central

Idiopathic Infantile Arterial Calcification is a rare autosome recessive disease characterized by extensive calcification of medium and large arteries. Loss-of-function mutations in ectonucleotide pyrophosphatase/phosphodiesterase 1 gene have been described in more than 80% of the cases. Although the diagnosis is usually made at autopsy, it is possible to identify cases based on clinical presentation, radiology findings, and molecular studies. Appropriate treatment can be initiated and has been shown to successfully induce permanent remission. We report a 4-week-old neonate who initially presented with respiratory distress, heart failure, and Coxsackie B viremia suggestive of viral induced cardiomyopathy. His symptoms progressed to multiple organ failure and he eventually expired at four weeks of age. On autopsy, diffuse calcium deposition within the internal elastic lamina of medium and large arteries was identified, as well as narrowing of lumen due to myointimal proliferation. This case report will emphasize the importance of taking this rare curable disease into consideration in all cases of infants with cardiopulmonary failure. PMID:24660083

Nael, A.; Siaghani, P. J.; Chen, D.; Romansky, S. G.; Shane, L.



Interleukin-6 receptor inhibition modulates the immune reaction and restores titin phosphorylation in experimental myocarditis.  


Increased levels of interleukin-6 (IL-6) have been observed in patients with acute myocarditis and are associated with poor prognosis. This study was designed to examine whether treatment with anti-IL-6 receptor antibody improves cardiac dysfunction and left ventricular (LV) remodeling in experimental Coxsackie virus B3 (CVB3)-induced myocarditis. C57BL6/J mice were subjected to acute CVB3 infection. One day after viral infection mice were treated with a single injection of an anti-IL-6 receptor antibody (MR16-1, tocilizumab) or control IgG. Seven days after viral infection, LV function was examined by conductance catheter technique, cardiac remodeling assessed by estimation of titin phosphorylation, cardiac fibrosis, and inflammatory and antiviral response by immunohistochemistry, RT-PCR and cell culture experiments. Compared to controls, infected mice displayed an impaired systolic and diastolic LV function associated with an increase in cardiac inflammation, fibrosis and impaired titin phosphorylation. IL-6 receptor blockade led to a shift of the immune response to a Th1 direction and significant reduction of viral load. In addition, cardiac immune response, extracellular matrix regulation and titin function improved, resulting in a preserved LV function. IL-6 receptor blockade exerts cardiac beneficial effects by antiviral and immunomodulatory actions after induction of an acute murine CVB3 virus myocarditis. PMID:25344085

Savvatis, Konstantinos; Müller, Irene; Fröhlich, Matthias; Pappritz, Kathleen; Zietsch, Christin; Hamdani, Nazha; Grote, Karsten; Schieffer, Bernhard; Klingel, Karin; Van Linthout, Sophie; Linke, Wolfgang A; Schultheiss, Heinz-Peter; Tschöpe, Carsten



Enhanced prostate cancer gene transfer and therapy using a novel serotype chimera cancer terminator virus (Ad.5/3-CTV).  


Few options are available for treating patients with advanced prostate cancer (PC). As PC is a slow growing disease and accessible by ultrasound, gene therapy could provide a viable option for this neoplasm. Conditionally replication-competent adenoviruses (CRCAs) represent potentially useful reagents for treating PC. We previously constructed a CRCA, cancer terminator virus (CTV), which showed efficacy both in vitro and in vivo for PC. The CTV was generated on a serotype 5-background (Ad.5-CTV) with infectivity depending on Coxsackie-Adenovirus Receptors (CARs). CARs are frequently reduced in many tumor types, including PCs thereby limiting effective Ad-mediated therapy. Using serotype chimerism, a novel CTV (Ad.5/3-CTV) was created by replacing the Ad.5 fiber knob with the Ad.3 fiber knob thereby facilitating infection in a CAR-independent manner. We evaluated Ad.5/3-CTV in comparison with Ad.5-CTV in low CAR human PC cells, demonstrating higher efficiency in inhibiting cell viability in vitro. Moreover, Ad.5/3-CTV potently suppressed in vivo tumor growth in a nude mouse xenograft model and in a spontaneously induced PC that develops in Hi-myc transgenic mice. Considering the significant responses in a Phase I clinical trial of a non-replicating Ad.5-mda-7 in advanced cancers, Ad.5/3-CTV may exert improved therapeutic benefit in a clinical setting. PMID:23868767

Azab, Belal M; Dash, Rupesh; Das, Swadesh K; Bhutia, Sujit K; Sarkar, Siddik; Shen, Xue-Ning; Quinn, Bridget A; Dent, Paul; Dmitriev, Igor P; Wang, Xiang-Yang; Curiel, David T; Pellecchia, Maurizio; Reed, John C; Sarkar, Devanand; Fisher, Paul B



Human enterovirus 71 epidemics: what's next?  

PubMed Central

Human enterovirus 71 (EV71) epidemics have affected various countries in the past 40 years. EV71 commonly causes hand, foot and mouth disease (HFMD) in children, but can result in neurological and cardiorespiratory complications in severe cases. Genotypic changes of EV71 have been observed in different places over time, with the emergence of novel genotypes or subgenotypes giving rise to serious outbreaks. Since the late 1990s, intra- and inter-typic recombination events in EV71 have been increasingly reported in the Asia-Pacific region. In particular, ‘double-recombinant’ EV71 strains belonging to a novel genotype D have been predominant in mainland China and Hong Kong over the last decade, though co-circulating with a minority of other EV71 subgenotypes and coxsackie A viruses. Continuous surveillance and genome studies are important to detect potential novel mutants or recombinants in the near future. Rapid and sensitive molecular detection of EV71 is of paramount importance in anticipating and combating EV71 outbreaks. PMID:24119538

Yip, Cyril C. Y.; Lau, Susanna K. P.; Woo, Patrick C. Y.; Yuen, Kwok-Yung



Cleavage of eukaryotic initiation factor eIF5B by enterovirus 3C proteases.  


The enteroviruses poliovirus (PV), Coxsackie B virus (CVB) and rhinovirus (HRV) are members of Picornaviridae that inhibit host cell translation early in infection. Enterovirus translation soon predominates in infected cells, but eventually also shuts off. This complex pattern of modulation of translation suggests regulation by a multifactorial mechanism. We report here that eIF5B is proteolytically cleaved during PV and CVB infection of cultured cells, beginning at 3 hours post-infection and increasing thereafter. Recombinant PV, CVB and HRV 3Cpro cleaved purified native rabbit eukaryotic initiation factor (eIF) 5B in vitro at a single site (VVEQG, equivalent to VMEQG479 in human eIF5B) that is consistent with the cleavage specificity of enterovirus 3C proteases. Cleavage separates the N-terminal domain of eIF5B from its essential conserved central GTPase and C-terminal domains. 3Cpro-mediated cleavage of eIF5B may thus play an accessory role in the shutoff of translation that occurs in enterovirus-infected cells. PMID:18572216

de Breyne, Sylvain; Bonderoff, Jennifer M; Chumakov, Konstantin M; Lloyd, Richard E; Hellen, Christopher U T



[Factors causing damage and destruction of beta-cells of the islets of Langerhans in the pancreas].  


Insulin secretion in patients with manifested diabetes mellitus tends to disappear months to decades after the diagnosis, which is a clear sign of a gradual loss of pancreatic islet beta-cells. In our sample of 30 type 2 diabetic patients, whose disease manifested between 30 and 45 years of age, about a half have retained or even increased insulin secretion 30 years later, while the other half exhibit a much diminished or lost insulin secretion. Factors that can damage or destroy beta-cells can be divided into the following groups: Metabolic factors: hyperglycemia and glucotoxicity, lipotoxicity, hypoxia, reactive oxygen species; Pharmacological factors: antimicrobial medication pentamidine, SSRI antidepressants; Factors related to impaired insulin secretion: MODY type diabetes; Environmental toxic factors: rat poison Vacor, streptozotocin, polychlorinated and polybrominated hydrocarbons; Disorders of the exocrine pancreas: tumor infiltration, fibrous infiltration, chronic pancreatitis, cystic fibrosis; Infections, inflammation, autoimmunity, viral factors: Coxsackie viruses, H1N1 influenza, enteroviruses. We are currently working on finding other factors leading to beta-cell damage, studying their effect on apoptosis and necrosis and looking for possible protective factors to prevent this damage. We our increasing knowledge about the mechanisms of beta-cell damage and destruction we come ever closer to suggest measures for their prevention. In this review we offer a brief and simplified summary of some of the findings related to this area.Key words: pancreatic islet beta-cells of Langerhans - factors damaging or destroying beta-cells - insulin secretion. PMID:25294754

And?l, Michal; N?mcová, Vlasta; Pavlíková, Nela; Urbanová, Jana; Cecháková, Marie; Havlová, Andrea; Straková, Radka; Ve?e?ová, Livia; Mandys, Václav; Ková?, Jan; Heneberg, Petr; Trnka, Jan; Polák, Jan



Paleobiogeography of the Onondaga Limestone in southeastern New York: A second shelf to basin ramp  

SciTech Connect

As a logical consequence of outcrop pattern and density, research on the Middle Devonian Onondaga Limestone has concentrated on the formation in an essentially east-west transect between the mid-Hudson Valley and Buffalo. Paleogeographic analysis reveals a symmetrical pattern, with the axis of the Appalachian Basin in central New York and the Edgecliff reefs flourishing in the eastern and western shelf regions. The subsurface reef trend of western New York and Pennsylvania is consistent with this model and is suggestive of a parallel reef trend in the east. However, recent examination of the formation in the southeastern part of the state has led to a pronounced reinterpretation of Onondaga paleobiogeography in that area. The authors have determined that there was a shallow carbonate shelf in the Helderberg-Coxsackie areas, a thick accumulation of shelf-margin bryozoan bafflestone between Leeds and Saugerties, and an even thicker accumulation of sparse to packed biocalcisiltites deposited on a carbonate ramp dipping southward into the Port Jervis area. It appears that this ramp occupied the northern margin of a structural base depocenter, located to the east of and not directly related to the topographic basin of central New York, which was centered in the Tristates vicinity from the Late Silurian until the early Middle Devonian.

Lindemann, R.H. (Skidmore Coll., Saratoga Springs, NY (United States). Dept. of Geology); Feldman, H.R. (Touro Coll., New York, NY (United States). Biology Dept.)



The post-viral syndrome: a review  

PubMed Central

The post-viral syndrome is described and its aetiology is discussed. Many features of the syndrome point to hysteria and altered medical perception as causes but much evidence for organic disease is also presented. Current interest focuses on recent or persisting infection with Coxsackie viruses. A balanced view of the syndrome as a mixture of organic and psychiatric dysfunction is offered. Widely differing estimates of incidence are quoted, possibly owing to varying medical awareness of the syndrome. Many drug therapies have been tried without success and management of the post-viral syndrome is hampered by the reluctance of patients to accept psychiatric support once the diagnosis is known. Many names have been proposed for the syndrome, some implying a purely physical or purely psychogenic aetiology: post-viral syndrome is suggested as the most appropriate term. Increased awareness of the syndrome will lead to an increase in its diagnosis in general practice: the role of the Myalgic Encephalomyelitis Association in promoting a combined psychiatric and organic view of the disease among sufferers is emphasized. PMID:3320358

Archer, M.I.



Neural stem cell-derived exosomes mediate viral entry  

PubMed Central

Background Viruses enter host cells through interactions of viral ligands with cellular receptors. Viruses can also enter cells in a receptor-independent fashion. Mechanisms regarding the receptor-independent viral entry into cells have not been fully elucidated. Exosomal trafficking between cells may offer a mechanism by which viruses can enter cells. Methods To investigate the role of exosomes on cellular viral entry, we employed neural stem cell-derived exosomes and adenovirus type 5 (Ad5) for the proof-of-principle study. Results Exosomes significantly enhanced Ad5 entry in Coxsackie virus and adenovirus receptor (CAR)-deficient cells, in which Ad5 only had very limited entry. The exosomes were shown to contain T-cell immunoglobulin mucin protein 4 (TIM-4), which binds phosphatidylserine. Treatment with anti-TIM-4 antibody significantly blocked the exosome-mediated Ad5 entry. Conclusion Neural stem cell-derived exosomes mediated significant cellular entry of Ad5 in a receptor-independent fashion. This mediation may be hampered by an antibody specifically targeting TIM-4 on exosomes. This set of results will benefit further elucidation of virus/exosome pathways, which would contribute to reducing natural viral infection by developing therapeutic agents or vaccines. PMID:25364247

Sims, Brian; Gu, Linlin; Krendelchtchikov, Alexandre; Matthews, Qiana L



The system of fucoidans from the brown seaweed Dictyota dichotoma: chemical analysis and antiviral activity.  


Room-temperature acid (pH 2) extraction of Dictyota dichotoma thalli yielded 2.2% of sulfated polysaccharides. Further extraction with the same solvent at 70°C was conducted sequentially for nine times, with a total yield of 7.2%. Fucose was the main monosaccharide only in the room-temperature extract (EAR) and in the first 70°C extract (EAH1). The remaining fractions showed increasing amounts of mannose (the main neutral monosaccharide), xylose and uronic acids. Fractionation by means of cetrimide precipitation and redissolution in increasing sodium chloride solutions has allowed obtaining several subfractions from each extract. The fractions redissolved at lower NaCl concentrations have large amounts of uronic acids and lesser sulfate contents, whereas those redissolved at higher NaCl concentrations are heavily sulfated and have low uronic acid contents. For the fucose-rich extracts (EAR and EAH1), fractionation leads to uronoxylomannofucan-rich and galactofucan-rich fractions. The remaining extracts gave rise to complex mixtures, with mannose and uronic acid-rich polysaccharides. Moderate inhibitory effect against herpes virus (HSV-1) and Coxsackie virus (CVB3) were found for the galactofucan-rich fractions. Most of the other fractions were inactive against both viruses, although some xylomannan-rich fractions were also active against HSV-1. PMID:24299842

Rabanal, Melissa; Ponce, Nora M A; Navarro, Diego A; Gómez, Ricardo M; Stortz, Carlos A



High prevalence of enteroviral genomic sequences in myocardium from cases of endemic cardiomyopathy (Keshan disease) in China  

PubMed Central

OBJECTIVE—To verify the aetiological involvement of enterovirus and identify the viral genomic sequences in Keshan disease.?DESIGN—Formalin fixed, paraffin embedded myocardial necropsy tissue samples were collected in Keshan disease endemic regions. Fourteen cases with a histologically confirmed diagnosis of subacute or chronic Keshan disease were studied. Control tissue included 10 samples of myocardium from cases of cerebral trauma and one from accidental acid intoxication. One sample from a case of enteroviral myocarditis was used as a positive control. The presence of viral genomic RNA was investigated using an established reverse transcription nested polymerase chain reaction (PCR) coupled with direct nucleotide sequencing. Further investigations of PCR positive samples included in situ antigen detection or hybridisation to confirm positive results.?RESULTS—Nine of 14 myocardial samples from Keshan disease cases and the positive control were positive for the enteroviral RNA. All the controls were negative. Six of the PCR positive samples were investigated further by in situ enteroviral antigen or RNA detection and all were positive. DNA sequencing of six representative PCR products confirmed that they were homologous to the 5' non-translated region of enteroviral genomic RNA. Five had highest homology to coxsackievirus B genotypes and one was identical to poliovirus type 3.?CONCLUSIONS—These results support an aetiological role for enteroviral infection in Keshan disease. Nucleotide sequence data suggest that coxsackievirus B or coxsackie B like viruses are often involved in Keshan disease.???Keywords: enterovirus; coxsackievirus; cardiomyopathy; Keshan disease PMID:10814633

Li, Y; Peng, T; Yang, Y; Niu, C; Archard, L; Zhang, H



Detection and rapid differentiation of human enteroviruses in water sources by restriction enzyme analysis.  


The objective of this study was to assess the application and efficiency of molecular techniques for the detection and serotyping of enteroviruses from environmental water samples. Samples of water were collected at regular intervals upstream and downstream of an informal settlement. Techniques for the detection of enteroviruses included a reverse transcription polymerase chain reaction (RT-PCR), nested PCR (n-PCR) and Sabin-specific triplex PCR. A specific 297 bp fragment was amplified by the n-PCR and subjected to restriction enzyme (RE) analysis to differentiate between various serotypes of prototypical enteroviruses. Enteroviruses that gave inconclusive restriction patterns were typed by partial sequencing of the VP1 region. Results indicated a high incidence of enteroviruses, predominantly coxsackie B viruses. The results on polioviruses, as well as other enteroviruses, contributed valuable information on enteroviruses circulating in the community. The molecular approach described here proved suitable for the rapid, sensitive, specific and cost effective, simultaneous detection and typing of enteroviruses in water. PMID:11464758

Vivier, J C; Clay, C G; Grabow, W O



Homosecoiridoid alkaloids with amino acid units from the flower buds of Lonicera japonica.  


Nine new homosecoiridoid alkaloids, named lonijaposides O-W (1-9), along with 19 known compounds, were isolated from an aqueous extract of the flower buds of Lonicera japonica. Their structures and absolute configurations were determined by spectroscopic data analysis and chemical methods. Lonijaposides O-W have structural features that involve amino acid units sharing the N atom with a pyridinium (1-5) or nicotinic acid (6-9) moiety. The absolute configurations of the amino acid units were determined by oxidation of each pyridinium ring moiety with potassium ferricyanide, hydrolysis of the oxidation product, and Marfey's analysis of the hydrolysate. This procedure was validated by oxidizing and hydrolyzing synthetic model compounds. The phenylalanine units in compounds 4, 5, and 9 have the d-configuration, and the other amino acid units in 1-3 and 6-8 possess the l-configuration. Compounds 1, 4, 6, and 9 and the known compounds 3,4-di-O-caffeoylquinic acid, 3,5-di-O-caffeoylquinic acid, and 5'-O-methyladenosine exhibited antiviral activity against the influenza virus A/Hanfang/359/95 (H3N2) with IC50 values of 3.4-11.6 ?M, and 4 inhibited Coxsackie virus B3 replication with an IC50 value of 12.3 ?M. PMID:24279769

Yu, Yang; Zhu, Chenggen; Wang, Sujuan; Song, Weixia; Yang, Yongchun; Shi, Jiangong



Presence of the Coxsackievirus and Adenovirus Receptor (CAR) in human neoplasms: a multitumour array analysis  

PubMed Central

Background: The Coxsackie- and Adenovirus Receptor (CAR) has been assigned two crucial attributes in carcinomas: (a) involvement in the regulation of growth and dissemination and (b) binding for potentially therapeutic adenoviruses. However, data on CAR expression in cancer types are conflicting and several entities have not been analysed to date. Methods: The expression of CAR was assessed by immunohistochemical staining of tissue microarrays (TMA) containing 3714 specimens derived from 100 malignancies and from 273 normal control tissues. Results: The expression of CAR was detected in all normal organs, except in the brain. Expression levels, however, displayed a broad range from being barely detectable (for example, in the thymus) to high abundance expression (for example, in the liver and gastric mucosa). In malignancies, a high degree of variability was notable also, ranging from significantly elevated CAR expression (for example, in early stages of malignant transformation and several tumours of the female reproductive system) to decreased CAR expression (for example, in colon and prostate cancer types). Conclusion: Our results provide a comprehensive insight into CAR expression in neoplasms and indicate that CAR may offer a valuable target for adenovirus-based therapy in a subset of carcinomas. Furthermore, these data suggest that CAR may contribute to carcinogenesis in an entity-dependent manner. PMID:24022195

Reeh, M; Bockhorn, M; Gorgens, D; Vieth, M; Hoffmann, T; Simon, R; Izbicki, J R; Sauter, G; Schumacher, U; Anders, M



Viral myocarditis and dilated cardiomyopathy: mechanisms, manifestations, and management  

PubMed Central

Viral infection of the heart is relatively common and usually of little consequence. It can, however, lead to substantial cardiac damage and severe acute heart failure. It can also evolve into the progressive syndrome of chronic heart failure. Recent studies have gone some way towards unravelling the complex mechanisms underlying the heart muscle damage that occurs after viral infection. These studies have lent support to both immune and viral mediated (independent of an immune response) cardiac damage. Acute myocarditis can present in various ways, and it may be a cause of sudden death in an otherwise healthy young adult. New treatments for viral heart disease are awaited. In the meanwhile, the haemodynamic support of patients with acute left ventricular failure caused by myocarditis should be aggressive, to allow for the possibility of spontaneous recovery. Contemporary trials of treatment in chronic heart failure secondary to dilated cardiomyopathy support the use of angiotensin converting enzyme inhibitors, ? adrenoceptor blockers, and spironolactone in such patients.???Keywords: myocarditis; heart failure; coxsackie B virus; dilated cardiomyopathy PMID:11123385

Kearney, M; Cotton, J; Richardson, P; Shah, A



Serologic and immunologic responses in chronic fatigue syndrome with emphasis on the Epstein-Barr virus.  


Although patients with chronic fatigue syndrome (CFS) can be diagnosed by clinical criteria, the lack of specific laboratory criteria delays or prevents the diagnosis and contributes to the quasi-disease status of the syndrome. A resurgence of interest in the syndrome has followed reports suggesting that CFS may be associated with chronic active infection due to the Epstein-Barr virus. Analysis of reports to date shows that the mean titers of antibodies to viral capsid antigen and to early antigen are greater for patients with CFS than for healthy individuals; this is particularly evident in cases for which serial samples were tested. However, these differences do not prove the cause of CFS. Cell-mediated immune responses in patients with CFS vary from study to study, and the number and function of natural killer cells in those patients are the most variable factors. Rates of isolation of virus from saliva do not differ, but in one comparison study with a large number of subjects, more lymphocytes that contained virus were isolated from patients than from controls. Other viruses, such as the Coxsackie B virus, have been implicated as causes of CFS in studies from Great Britain. The use of a working definition of CFS and standardized tests to address abnormalities revealed by laboratory tests among homogeneous populations should allow determination of useful tests for the diagnosis of CFS and studies of its mechanisms. PMID:1850541

Jones, J F



Enterovirus related metabolic myopathy: a postviral fatigue syndrome  

PubMed Central

Objective: To detect and characterise enterovirus RNA in skeletal muscle from patients with chronic fatigue syndrome (CFS) and to compare efficiency of muscle energy metabolism in enterovirus positive and negative CFS patients. Methods: Quadriceps muscle biopsy samples from 48 patients with CFS were processed to detect enterovirus RNA by two stage, reverse transcription, nested polymerase chain reaction (RT-NPCR), using enterovirus group specific primer sets. Direct nucleotide sequencing of PCR products was used to characterise the enterovirus. Controls were 29 subjects with normal muscles. On the day of biopsy, each CFS patient undertook a subanaerobic threshold exercise test (SATET). Venous plasma lactate was measured immediately before and after exercise, and 30 minutes after testing. An abnormal lactate response to exercise (SATET+) was defined as an exercise test in which plasma lactate exceeded the upper 99% confidence limits for normal sedentary controls at two or more time points. Results: Muscle biopsy samples from 20.8% of the CFS patients were positive for enterovirus sequences by RT-NPCR, while all the 29 control samples were negative; 58.3% of the CFS patients had a SATET+ response. Nine of the 10 enterovirus positive cases were among the 28 SATET+ patients (32.1%), compared with only one (5%) of the 20 SATET- patients. PCR products were most closely related to coxsackie B virus. Conclusions: There is an association between abnormal lactate response to exercise, reflecting impaired muscle energy metabolism, and the presence of enterovirus sequences in muscle in a proportion of CFS patients. PMID:14570830

Lane, R; Soteriou, B; Zhang, H; Archard, L



In Vivo Bioluminescence Tumor Imaging of RGD Peptide-modified Adenoviral Vector Encoding Firefly Luciferase Reporter Gene  

PubMed Central

Purpose The goal of this study is to demonstrate the feasibility of chemically modified human adenovirus (Ad) vectors for tumor retargeting. Procedures E1- and E3-deleted Ad vectors carrying firefly luciferase reporter gene under cytomegalovirus promoter (AdLuc) was surface-modified with cyclic arginine–glycine–aspartic acid (RGD) peptides through a bifunctional poly(ethyleneglycol) linker (RGD-PEG-AdLuc) for integrin ?v?3 specific delivery. The Coxsackie and adenovirus viral receptor (CAR) and integrin ?v?3 expression in various tumor cell lines was determined by reverse transcriptase PCR and fluorescence-activated cell sorting. Bioluminescence imaging was performed in vitro and in vivo to evaluate RGD-modified AdLuc infectivity. Results RGD-PEG-AdLuc abrogated the native CAR tropism and exhibited significantly enhanced transduction efficiency of integrin-positive tumors than AdLuc through intravenous administration. Conclusion This approach provides a robust platform for site-specific gene delivery and noninvasive monitoring of the transgene delivery efficacy and homing. PMID:17297551

Niu, Gang; Xiong, Zhengming; Cheng, Zhen; Cai, Weibo; Gambhir, Sanjiv S.; Xing, Lei; Chen, Xiaoyuan



Adenoviral virotherapy for malignant brain tumors  

PubMed Central

Glioblastoma multiforme (GBM) is the most common form of primary brain cancer. In the past decade, virotherapy of tumors has gained credence, particularly in glioma management, as these tumors are not completely resectable and tend to micro-metastasize. Adenoviral vectors have an advantage over other viral vectors in that they are relatively non-toxic and do not integrate in the genome. However, the lack of coxsackie and adenovirus receptors (CAR) on surface of gliomas provides for inefficient transduction of wild-type adenoviral vectors in these tumors. By targeting receptors that are over-expressed in gliomas, modified adenoviral constructs have been shown to efficiently infect glioma cells. In addition, by taking advantage of tumor specific promoter (TSP) elements, oncolytic adenoviral vectors offer the promise of selective tumor-specific replication. This dual targeting strategy has enabled specificity in both laboratory and pre-clinical settings. This review looks at current trends in adenoviral virotherapy of gliomas, with an emphasis on targeting modalities and future clinical applications. PMID:19456208

Nandi, Suvobroto; Lesniak, Maciej S



Possible Involvement of Opa-Interacting Protein 5 in Adipose Proliferation and Obesity  

PubMed Central

Obesity is an epidemic matter increasing risk for cardiovascular diseases and metabolic disorders such as type 2 diabetes. We recently examined the association between visceral fat adiposity and gene expression profile of peripheral blood cells in human subjects. In a series of studies, Opa (Neisseria gonorrhoeae opacity-associated)-interacting protein 5 (OIP5) was nominated as a molecule of unknown function in adipocytes and thus the present study was performed to investigate the role of OIP5 in obesity. Adenovirus overexpressing Oip5 (Ad-Oip5) was generated and infected to 3T3-L1 cells stably expressing Coxsackie-Adenovirus Receptor (CAR-3T3-L1) and to mouse subcutaneous fat. For a knockdown experiment, siRNA against Oip5 (Oip5-siRNA) was introduced into 3T3-L1 cells. Proliferation of adipose cells was measured by BrdU uptake, EdU-staining, and cell count. Significant increase of Oip5 mRNA level was observed in obese white adipose tissues and such increase was detected in both mature adipocytes fraction and stromal vascular cell fraction. Ad-Oip5-infected CAR-3T3-L1 preadipocytes and adipocytes proliferated rapidly, while a significant reduction of proliferation was observed in Oip5-siRNA-introduced 3T3-L1 preadipocytes. Fat weight and number of adipocytes were significantly increased in Ad-Oip5-administered fat tissues. Oip5 promotes proliferation of pre- and mature-adipocytes and contributes adipose hyperplasia. Increase of Oip5 may associate with development of obesity. PMID:24516558

Inoue, Kana; Maeda, Norikazu; Mori, Takuya; Sekimoto, Ryohei; Tsushima, Yu; Matsuda, Keisuke; Yamaoka, Masaya; Suganami, Takayoshi; Nishizawa, Hitoshi; Ogawa, Yoshihiro; Funahashi, Tohru; Shimomura, Iichiro



A pathogenic mechanism of chronic ongoing myocarditis.  


To clarify the pathogenetic mechanism of chronic ongoing myocarditis, we produced Coxsackievirus B3-induced myocarditis in A/J mice and immunopathologically examined the microcirculation in the chronic phase of myocarditis. Forty-two 3-week-old A/J mice were inoculated intraperitoneally with Coxsackievirus B3 (Nancy strain) 2 x 10(4) PFU (plaque-forming units) and sacrificed 7, 14, 21, 50, 90, or 120 days later. To evaluate myocardial microcirculation, 18 of the hearts were perfused from the thoracic aorta with warm 2% gelatin/carbon solution. The remaining hearts were quickly frozen for immunologic analysis with an enzyme immunostaining assay using monoclonal antibodies against CD4, CD8, macrophages, intercellular adhesion molecule-1 (ICAM-1) and major histocompatibility complex class I or II. The presence of viral RNA genome in the myocardium at 40, 50, or 60 days after inoculation was evaluated using the polymerase chain reaction. The lesions in chronic ongoing myocarditis consisted of myocardial damage, myocardial calcification, interstitial fibrosis, and infiltration of mononuclear cells. These infiltrated lymphocytes were predominantly CD4+ T cells. Furthermore, microvascular abnormalities, including dilatation, tortuosity, constriction, and abrupt termination, were observed around the lesions. There was marked infiltration by mononuclear cells around the microvessels. ICAM-1 was strongly expressed in the endothelial cells of the vessels. Coxsackie B3 viral genome was not detected in the myocardium of mice with chronic ongoing myocarditis in each stage examined. These results suggest that an autoimmune mechanism is involved in the persistent inflammation seen in chronic ongoing myocarditis. PMID:8889664

Nakamura, H; Yamamura, T; Fukuta, S; Matsumori, A; Matsuzaki, M



Identification of novel small molecule inhibitors of adenovirus gene transfer using a high throughput screening approach.  


Due to many favourable attributes adenoviruses (Ads) are the most extensively used vectors for clinical gene therapy applications. However, following intravascular administration, the safety and efficacy of Ad vectors are hampered by the strong hepatic tropism and induction of a potent immune response. Such effects are determined by a range of complex interactions including those with neutralising antibodies, blood cells and factors, as well as binding to native cellular receptors (coxsackie adenovirus receptor (CAR), integrins). Once in the bloodstream, coagulation factor X (FX) has a pivotal role in determining Ad liver transduction and viral immune recognition. Due to difficulties in generating a vector devoid of multiple receptor binding motifs, we hypothesised that a small molecule inhibitor would be of value. Here, a pharmacological approach was implemented to block adenovirus transduction pathways. We developed a high throughput screening (HTS) platform to identify small molecule inhibitors of FX-mediated Ad5 gene transfer. Using an in vitro fluorescence and cell-based HTS, we evaluated 10,240 small molecules. Following sequential rounds of screening, three compounds, T5424837, T5550585 and T5660138 were identified that ablated FX-mediated Ad5 transduction with low micromolar potency. The candidate molecules possessed common structural features and formed part of the one pharmacophore model. Focused, mini-libraries were generated with structurally related molecules and in vitro screening revealed novel hits with similar or improved efficacy. The compounds did not interfere with Ad5:FX engagement but acted at a subsequent step by blocking efficient intracellular transport of the virus. In vivo, T5660138 and its closely related analogue T5660136 significantly reduced Ad5 liver transgene expression at 48 h post-intravenous administration of a high viral dose (1×10¹¹ vp/mouse). Therefore, this study identifies novel and potent small molecule inhibitors of the Ad5 transduction which may have applications in the Ad gene therapy setting. PMID:23702233

Duffy, Margaret R; Parker, Alan L; Kalkman, Eric R; White, Katie; Kovalskyy, Dmytro; Kelly, Sharon M; Baker, Andrew H



mRNA translation is compartmentalized to the endoplasmic reticulum following physiological inhibition of cap-dependent translation  

PubMed Central

Eukaryotic cells utilize a cycle of ribosome trafficking on the endoplasmic reticulum (ER) to partition mRNAs between the cytosol and ER compartments. In this process, ribosomes engaged in the synthesis of signal sequence-bearing proteins are trafficked to the endoplasmic reticulum via the signal-recognition particle pathway and are released from the ER upon translation termination. Though the processes governing ribosome trafficking to the ER are well understood, little is known regarding the complementary ribosome release process. In this study, Coxsackie B virus (CBV) infection was used to inactivate the initiation stage of protein synthesis, thereby limiting translation to the elongation and termination stages. Ribosome partitioning between the cytosol and ER compartments was examined to determine the role of termination in ribosome release from the ER. CBV infection resulted in efficient cleavage of eIF4G and PABP, coincident with polyribosome breakdown in the cytosol and ER compartments. Termination resulted in the continued association of ribosomes with the ER compartment, rather than the expected process of ribosome release. Analyses of ribosome/mRNA loading patterns in the cytosol and ER revealed that CBV infection was accompanied by a suppression of mRNA translation in the cytosol and the sustained, although reduced, translation in the ER compartment. Direct biosynthetic labeling experiments demonstrated that protein synthesis on the ER was enhanced relative to the cytosol following CBV infection. In total, these data demonstrate that ribosome and mRNA release from the ER is regulated independent of translation termination and identify the ER as a privileged site for protein synthesis. PMID:16540694

Lerner, Rachel S.; Nicchitta, Christopher V.



Animal Models used to Examine the Role of the Environment in the Development of Autoimmune Disease: Findings from an NIEHS Expert Panel Workshopa  

PubMed Central

Autoimmunity is thought to result from a combination of genetics, environmental triggers, and stochastic events. Environmental factors, such as chemicals, drugs or infectious agents, have been implicated in the expression of autoimmune disease, yet human studies are extremely limited in their ability to test isolated exposures to demonstrate causation or to assess pathogenic mechanisms. In this review we examine the research literature on the ability of chemical, physical and biological agents to induce and/or exacerbate autoimmunity in a variety of animal models. There is no single animal model capable of mimicking the features of human autoimmune disease, particularly as related to environmental exposures. An objective, therefore, was to assess the types of information that can be gleaned from the use of animal models, and how well that information can be used to translate back to human health. Our review notes the importance of genetic background to the types and severity of the autoimmune response following exposure to environmental factors, and emphasizes literature where animal model studies have led to increased confidence about environmental factors that affect expression of autoimmunity. A high level of confidence was reached if there were multiple studies from different laboratories confirming the same findings. Examples include mercury, pristane, and infection with Streptococcus or Coxsackie B virus. A second level of consensus identified those exposures likely to influence autoimmunity but requiring further confirmation. To fit into this category, there needed to be significant supporting data, perhaps by multiple studies from a single laboratory, or repetition of some but not all findings in multiple laboratories. Examples include silica, gold, TCE, TCDD, UV radiation, and Theiler’s murine encephalomyelitis virus. With the caveat that researchers must keep in mind the limitations and appropriate applications of the various approaches, animal models are shown to be extremely valuable tools for studying the induction or exacerbation of autoimmunity by environmental conditions and exposures. PMID:22748431

Germolec, Dori; Kono, Dwight H.; Pfau, Jean C.; Pollard, K. Michael



“Bath salts” induced severe reversible cardiomyopathy  

PubMed Central

Patient: Male, 27 Final Diagnosis: Bath salt induced cardiomyopathy Symptoms: Agitation • fever • pedal edema Medication: Intravenous nor-epinephrine for less than 6 hours Clinical Procedure: — Specialty: Internal medicine • cardiology Objective: Unusual clinical course Background: “Bath salts” is the street name for a group of recently identified and increasingly abused stimulant synthetic cathinones that are associated with multiple systemic effects. We present a case of a patient who developed reversible dilated cardiomyopathy secondary to their use. Case Report: A 27 year old male with no past medical history was brought to emergency department with agitation. He had been inhaling and intravenously injecting “bath salts”, containing a mephedrone/Methylenedioxypyrovalerone (MDPV) combination. On presentation, he was tachycardic, hypotensive and febrile. His initial labs showed an elevated white count, creatinine and creatinine phosphokinase levels. His erythrocyte sedimentation rate; C-reactive protein; urinalysis; urine drug screen; Human Immunodeficiency Virus, hepatitis, coxsackie, and influenza serology were normal. EKG showed sinus tachycardia. An echocardiogram was done which showed dilated cardiomyopathy with an ejection fraction (EF) of 15–20% and global hypokinesia. A left heart catheterization was done and was negative for coronary artery disease. At a 20 week follow up, he had stopped abusing bath salts and was asymptomatic. A repeat echocardiogram showed an EF of 52%. Cocnlusions: Bath salts (MDPV, mephedrone) are synthetic cathinones with amphetamine/cocaine like properties with potential cardiotoxic effects. Cardiovascular manifestations reported include tachycardia, hypertension, myocardial infarction, arrhythmias and cardiac arrest. “Bath salts” can also cause severe reversible dilated cardiomyopathy. Prior to diagnosis, other causes of cardiomyopathy including ischemic, infectious, familial, immunological, metabolic and cytotoxic may need to be ruled out; as was done in our patient. PMID:23919103

Sivagnanam, Kamesh; Chaudari, Dhara; Lopez, Pablo; Sutherland, Michael E; Ramu, Vijay K.



The hemorrhagic fevers of Southern Africa with special reference to studies in the South African Institute for Medical Research.  


In this review of studies on the hemorrhagic fevers of Southern Africa carried out in the South African Institute for Medical Research, attention has been called to occurrence of meningococcal septicemia in recruits to the mining industry and South African Army, to cases of staphylococcal and streptococcal septicemia with hemorrhagic manifestations, and to the occurrence of plague which, in its septicemic form, may cause a hemorrhagic state. "Onyalai," a bleeding disease in tropical Africa, often fatal, was related to profound thrombocytopenia possibly following administration of toxic witch doctor medicine. Spirochetal diseases, and rickettsial diseases in their severe forms, are often manifested with hemorrhagic complications. Of enterovirus infections, Coxsackie B viruses occasionally caused severe hepatitis associated with bleeding, especially in newborn babies. Cases of hemorrhagic fever presenting in February-March, 1975 are described. The first outbreak was due to Marburg virus disease and the second, which included seven fatal cases, was caused by Rift Valley fever virus. In recent cases of hemorrhagic fever a variety of infective organisms have been incriminated including bacterial infections, rickettsial diseases, and virus diseases, including Herpesvirus hominis; in one patient, the hemorrhagic state was related to rubella. A boy who died in a hemorrhagic state was found to have Congo fever; another patient who died of severe bleeding from the lungs was infected with Leptospira canicola, and two patients who developed a hemorrhagic state after a safari trip in Northern Botswana were infected with Trypanosoma rhodesiense. An illness manifested by high fever and melena developed in a young man after a visit to Zimbabwe; the patient was found to have both malaria and Marburg virus disease. PMID:6897472

Gear, J H



Antiviral activity and mode of action of caffeoylquinic acids from Schefflera heptaphylla (L.) Frodin.  


Schefflera heptaphylla is a popular medicinal plant in southern China. Three caffeoylquinic acid derivatives, namely 3,4-di-O-caffeoylquinic acid, 3,5-di-O-caffeoylquinic acid, and 3-O-caffeoylquinic acid, were isolated from this plant and investigated for their antiviral activity against respiratory syncytial virus (RSV). 3,4-Di-O-caffeoylquinic acid and 3,5-di-O-caffeoylquinic acid possessed potent anti-RSV activity. The median inhibitory concentrations (IC50) of 3,4-di-O-caffeoylquinic acid and 3,5-di-O-caffeoylquinic acid against RSV were 2.33 microM (1.2 microg/ml) and 1.16 microM (0.6 microg/ml), respectively, in a plaque reduction assay. The dicaffeoylquinic acids exhibited minimal cytotoxicity against HEp-2 cells with median cytotoxic concentration (CC50) higher than 1000 microM. The maximal non-cytotoxic concentration (MNCC) of the two dicaffeoylquinic acids were about 96.7 microM, which suggested their anti-RSV effect was not due to cytotoxicity. The antiviral action of 3,4-di-O-caffeoylquinic acid and 3,5-di-O-caffeoylquinic acid was specific against RSV, as they had no obvious antiviral activity against influenza A (Flu A), Coxsackie B3 (Cox B3), and Herpes simplex type one (HSV-1) viruses. Studies were performed that indicated that the dicaffeoylquinic acids could inhibit RSV directly, extracellularly, but only at much higher concentrations than seen in standard assays. Moreover, they could not inhibit RSV attachment to host cells, and could not protect HEp-2 cells from RSV infection at lower concentrations. The data suggest that the compounds exerted their anti-RSV effects via the inhibition of virus-cell fusion in the early stage, and the inhibition of cell-cell fusion at the end of the RSV replication cycle. PMID:16140400

Li, Yaolan; But, Paul P H; Ooi, Vincent E C



Primary Murine CD4+ T Cells Fail to Acquire the Ability to Produce Effector Cytokines When Active Ras Is Present during Th1/Th2 Differentiation  

PubMed Central

Constitutive Ras signaling has been shown to augment IL-2 production, reverse anergy, and functionally replace many aspects of CD28 co-stimulation in CD4+ T cells. These data raise the possibility that introduction of active Ras into primary T cells might result in improved functionality in pathologic situations of T cell dysfunction, such as cancer or chronic viral infection. To test the biologic effects of active Ras in primary T cells, CD4+ T cells from Coxsackie-Adenovirus Receptor Transgenic mice were transduced with an adenovirus encoding active Ras. As expected, active Ras augmented IL-2 production in naive CD4+ T cells. However, when cells were cultured for 4 days under conditions to promote effector cell differentiation, active Ras inhibited the ability of CD4+ T cells to acquire a Th1 or Th2 effector cytokine profile. This differentiation defect was not due to deficient STAT4 or STAT6 activation by IL-12 or IL-4, respectively, nor was it associated with deficient induction of T-bet and GATA-3 expression. Impaired effector cytokine production in active Ras-transduced cells was associated with deficient demethylation of the IL-4 gene locus. Our results indicate that, despite augmenting acute activation of naïve T cells, constitutive Ras signaling inhibits the ability of CD4+ T cells to properly differentiate into Th1/Th2 effector cytokine-producing cells, in part by interfering with epigenetic modification of effector gene loci. Alternative strategies to potentiate Ras pathway signaling in T cells in a more regulated fashion should be considered as a therapeutic approach to improve immune responses in vivo. PMID:25397617

Janardhan, Sujit V.; Marks, Reinhard; Gajewski, Thomas F.



Species D Human Adenovirus Type 9 Exhibits Better Virus-Spread Ability for Antitumor Efficacy among Alternative Serotypes  

PubMed Central

Species C human adenovirus serotype 5 (HAdV-C5) is widely used as a vector for cancer gene therapy, because it efficiently transduces target cells. A variety of HAdV-C5 vectors have been developed and tested in vitro and in vivo for cancer gene therapy. While clinical trials with HAdV-C5 vectors resulted in effective responses in many cancer patients, administration of HAdV-C5 vectors to solid tumors showed responses in a limited area. A biological barrier in tumor mass is considered to hinder viral spread of HAdV-C5 vectors from infected cells. Therefore, efficient virus-spread from an infected tumor cell to surrounding tumor cells is required for successful cancer gene therapy. In this study, we compared HAdV-C5 to sixteen other HAdV serotypes selected from species A to G for virus-spread ability in vitro. HAdV-D9 showed better virus-spread ability than other serotypes, and its viral progeny were efficiently released from infected cells during viral replication. Although the HAdV-D9 fiber protein contains a binding site for coxsackie B virus and adenovirus receptor (CAR), HAdV-D9 showed expanded tropism for infection due to human CAR (hCAR)-independent attachment to target cells. HAdV-D9 infection effectively killed hCAR-negative cancer cells as well as hCAR-positive cancer cells. These results suggest that HADV-D9, with its better virus-spread ability, could have improved therapeutic efficacy in solid tumors compared to HAdV-C5. PMID:24503714

Uchino, Junji; Curiel, David T.; Ugai, Hideyo



Animal models used to examine the role of the environment in the development of autoimmune disease: findings from an NIEHS Expert Panel Workshop.  


Autoimmunity is thought to result from a combination of genetics, environmental triggers, and stochastic events. Environmental factors, such as chemicals, drugs or infectious agents, have been implicated in the expression of autoimmune disease, yet human studies are extremely limited in their ability to test isolated exposures to demonstrate causation or to assess pathogenic mechanisms. In this review we examine the research literature on the ability of chemical, physical and biological agents to induce and/or exacerbate autoimmunity in a variety of animal models. There is no single animal model capable of mimicking the features of human autoimmune disease, particularly as related to environmental exposures. An objective, therefore, was to assess the types of information that can be gleaned from the use of animal models, and how well that information can be used to translate back to human health. Our review notes the importance of genetic background to the types and severity of the autoimmune response following exposure to environmental factors, and emphasizes literature where animal model studies have led to increased confidence about environmental factors that affect expression of autoimmunity. A high level of confidence was reached if there were multiple studies from different laboratories confirming the same findings. Examples include mercury, pristane, and infection with Streptococcus or Coxsackie B virus. A second level of consensus identified those exposures likely to influence autoimmunity but requiring further confirmation. To fit into this category, there needed to be significant supporting data, perhaps by multiple studies from a single laboratory, or repetition of some but not all findings in multiple laboratories. Examples include silica, gold, TCE, TCDD, UV radiation, and Theiler's murine encephalomyelitis virus. With the caveat that researchers must keep in mind the limitations and appropriate applications of the various approaches, animal models are shown to be extremely valuable tools for studying the induction or exacerbation of autoimmunity by environmental conditions and exposures. PMID:22748431

Germolec, Dori; Kono, Dwight H; Pfau, Jean C; Pollard, K Michael



MiR-155-mediated loss of C/EBP? shifts the TGF-? response from growth inhibition to epithelial-mesenchymal transition, invasion and metastasis in breast cancer  

PubMed Central

During breast cancer progression, transforming growth factor-beta (TGF-?) switches from acting as a growth inhibitor to become a major promoter of epithelial-mesenchymal transition (EMT), invasion and metastasis. However, the mechanisms involved in this switch are not clear. We found that loss of CCAAT-enhancer binding protein beta (C/EBP?), a differentiation factor for the mammary epithelium, was associated with signs of EMT in triple-negative human breast cancer, and in invasive areas of mammary tumors in MMTV-PyMT mice. Using an established model of TGF-?-induced EMT in mouse mammary gland epithelial cells, we discovered that C/EBP? was repressed during EMT by miR-155, an oncomiR in breast cancer. Depletion of C/EBP? potentiated the TGF-? response towards EMT, and contributed to evasion of the growth inhibitory response to TGF-?. Furthermore, loss of C/EBP? enhanced invasion and metastatic dissemination of the mouse mammary tumor cells to the lungs after subcutaneous injection into mice. The mechanism by which loss of C/EBP? promoted the TGF-? response towards EMT, invasion and metastasis, was traced to a previously uncharacterized role of C/EBP? as a transcriptional activator of genes encoding the epithelial junction proteins E-cadherin and coxsackie virus and adenovirus receptor. The results identify miR-155-mediated loss of C/EBP? as a mechanism, which promotes breast cancer progression by shifting the TGF-? response from growth inhibition to EMT, invasion and metastasis. PMID:23955085

Johansson, J; Berg, T; Kurzejamska, E; Pang, M-F; Tabor, V; Jansson, M; Roswall, P; Pietras, K; Sund, M; Religa, P; Fuxe, J



The combination of an oxygen-dependent degradation domain-regulated adenovirus expressing the chemokine RANTES/CCL5 and NK-92 cells exerts enhanced antitumor activity in hepatocellular carcinoma  

PubMed Central

Oncolytic adenoviruses are modified based on adenovirus serotype 5 (Ad5), which belongs to subgroup C and depends on Coxsackie-adenovirus receptor (CAR) to recognize target cells. However, expression of CAR is generally low or lost in certain tumors including hepatocellular carcinoma (HCC). By contrast, CD46 is highly expressed in various types of malignant tumor cells. Therefore, we constructed an adenovirus vector expressing the human RANTES/CCL5 gene regulated by oxygen-dependent degradation domain (ODD) and analyzed its antitumor effects in vitro and in vivo. The human RANTES/CCL5 gene was fused with ODD by PCR and the recombinant oncolytic adenovirus containing RANTES-ODD, SG511-CCL5-ODD, was constructed by the Gateway system, which infected cells by binding CD46. Viral replication experiments were performed to evaluate the selective replication ability of SG511-CCL5-ODD. RANTES expression was determined by ELISA. The chemotactic test was used to analyze the ability of the expressed RANTES to recruit NK92 cells. The antitumor effects of SG511-CCL5-ODD were examined in HCC xenografts in nude mice. A chimeric oncolytic adenovirus, SG511-CCL5-ODD, was constructed successfully. Cells infected with the recombinant virus were able to express RANTES selectively in different environments controlled by ODD and the expressed RANTES was able to recruit NK92 cells by its chemotactic effect in vitro and improve the anticancer immune response in HCC xenografts in nude mice. The chimeric adenovirus SG511-CCL5-ODD highly expressed the RANTES-ODD fusion gene in the hypoxia of HCC under the control of the ODD and effectively attracted NK92 cells and a high number of immunocytes. These factors had complementary advantages and, in combination, exerted enhanced antitumor efficacy. PMID:23292657




Increased Adenovirus Type 5 Mediated Transgene Expression Due to RhoB Down-Regulation  

PubMed Central

Adenovirus type 5 (Ad5) is a non-enveloped DNA virus frequently used as a gene transfer vector. Efficient Ad5 cell entry depends on the availability of its primary receptor, coxsackie and adenovirus receptor, which is responsible for attachment, and integrins, secondary receptors responsible for adenovirus internalization via clathrin-mediated endocytosis. However, efficacious adenovirus-mediated transgene expression also depends on successful trafficking of Ad5 particles to the nucleus of the target cell. It has been shown that changes occurring in tumor cells during development of resistance to anticancer drugs can be beneficial for adenovirus mediated transgene expression. In this study, using an in vitro model consisting of a parental cell line, human laryngeal carcinoma HEp2 cells, and a cisplatin-resistant clone CK2, we investigated the cause of increased Ad5-mediated transgene expression in CK2 as compared to HEp2 cells. We show that the primary cause of increased Ad5-mediated transgene expression in CK2 cells is not modulation of receptors on the cell surface or change in Ad5wt attachment and/or internalization, but is rather the consequence of decreased RhoB expression. We propose that RhoB plays an important role in Ad5 post-internalization events and more particularly in Ad5 intracellular trafficking. To the best of our knowledge, this is the first study showing changed Ad5 trafficking pattern between cells expressing different amount of RhoB, indicating the role of RhoB in Ad5 intracellular trafficking. PMID:24466204

Majhen, Dragomira; Stojanovic, Nikolina; Vukic, Dunja; Pichon, Chantal; Leduc, Chloe; Osmak, Maja; Ambriovic-Ristov, Andreja



A hexon and fiber-modified adenovirus expressing CD40L improves the antigen presentation capacity of dendritic cells.  


CD40 ligand (CD40L), a strong stimulator of Th1 immune responses, acts via dendritic cells to trigger T-cell activation. AdCD40L therapy introduces the CD40L gene into the tumor microenvironment with an adenoviral vector and has shown promising results in experimental tumor models, dogs, and patients (phase I-II trials). The transduction efficiency of AdCD40L is dependent on the expression of CAR (coxsackie/adenovirus adhesion receptor), which is commonly downregulated on tumor cells. To enhance transduction efficiency, and therefore the therapeutic efficacy, a double-modified adenovirus was developed. The double-modified Ad5PTDf35(mCD40L) had a protein transduction domain (PTD) inserted into the hexon protein and the virus fiber is switched from serotype 5 to serotype 35. These modifications enable transduction of a wider range of cell types. In comparison with Ad5(mCD40L), Ad5PTDf35(mCD40L) showed increased transduction capacity on a variety of murine cells. Furthermore, antigen presentation was improved after transduction with Ad5PTDf35(mCD40L). This was demonstrated in an antigen presentation assay, both in vitro and in vivo, in which transduced dendritic cells were loaded with suboptimal concentrations of the human gp100 peptide and allowed to interact with gp100-specific transgenic T cells (pmel). Finally, Ad5PTDf35(mCD40L) could delay tumor growth in a murine cancer model at a particle load, wherein therapeutic efficacy of the Ad5(mCD40L) vector was lost. Hence, the Ad5PTDf35(CD40L) vector holds great promise as a second-generation immune stimulatory therapy, as it not only targets tumor cells but also antigen-presenting cells that are, among other cells, present in the tumor microenvironment. PMID:24598450

Liljenfeldt, Lina; Yu, Di; Chen, Liye; Essand, Magnus; Mangsbo, Sara M



Prevalence of Nonpolio Enteroviruses in the Sewage of Guangzhou City, China, from 2009 to 2012  

PubMed Central

The human-pathogenic viruses in urban sewage have been extensively monitored to obtain information on circulating viruses in human communities. Enteroviruses (EVs) excreted by patients who present with diverse clinical syndromes can remain infectious in the environment for several weeks, and limited data on circulating environmental EVs are available. A 4-year (2009 to 2012) surveillance study was conducted to detect nonpolio enteroviruses (NPEVs) in the urban sewage of Guangzhou city, China. After the viruses in the sewage samples were concentrated and isolated, molecular identification was used to detect and type the NPEVs. During the 4-year study, 17 different NPEV serotypes were identified in the sewage of Guangzhou city. The most common serotypes were echovirus 11 (ECHO11), ECHO6, ECHO7, and ECHO12 and coxsackie group B viruses 5 (CVB5) and CVB3. The predominant serotypes were influenced by spatial and temporal factors and differed each year. CVB5 was commonly detected in 2009 and 2010 but was rarely isolated in 2011 and 2012. In contrast, CVB3 was not observed in 2009 and 2010 but was increasingly detected in 2011 and 2012. Our study provides an overview of the serotype distribution and circulation patterns of NPEVs in the sewage of Guangzhou, China. In the absence of a systematic EV disease surveillance system, the detection and characterization of sewage-borne NPEVs will help us better understand the changes in EV disease trends and the epidemic background of circulating EVs, which could help interpret the EV trends and warn of future outbreaks in this area. PMID:24096418

Lu, Jing; Zhang, Yong; Yoshida, Hiromu; Guo, Xue; Liu, Leng; Li, Hui; Zeng, Hanri; Fang, Ling; Mo, Yanling; Yi, Lina; Chosa, Toru; Xu, Wenbo; Ke, Changwen



Multimerization of Adenovirus Serotype 3 Fiber Knob Domains Is Required for Efficient Binding of Virus to Desmoglein 2 and Subsequent Opening of Epithelial Junctions?  

PubMed Central

Recently, we identified desmoglein 2 (DSG2) as the main receptor for a group of species B adenoviruses (Ads), including Ad3, a serotype that is widely distributed in the human population (H. Wang et al., Nat. Med. 17:96–104, 2011). In this study, we have attempted to delineate structural details of the Ad3 interaction with DSG2. For CAR- and CD46-interacting Ad serotypes, attachment to cells can be completely blocked by an excess of recombinant fiber knob protein, while soluble Ad3 fiber knob only inefficiently blocks Ad3 infection. We found that the DSG2-interacting domain(s) within Ad3 is formed by several fiber knob domains that have to be in the spatial constellation that is present in viral particles. Based on this finding, we generated a small recombinant, self-dimerizing protein containing the Ad3 fiber knob (Ad3-K/S/Kn). Ad3-K/S/Kn bound to DSG2 with high affinity and blocked Ad3 infection. We demonstrated by confocal immunofluorescence and transmission electron microscopy analyses that Ad3-K/S/Kn, through its binding to DSG2, triggered the transient opening of intercellular junctions in epithelial cells. The pretreatment of epithelial cells with Ad3-K/S/Kn resulted in increased access to receptors that are localized in or masked by epithelial junctions, e.g., CAR or Her2/neu. Ad3-K/S/Kn treatment released CAR from tight junctions and thus increased the transduction of epithelial cells by a serotype Ad5-based vector. Furthermore, the pretreatment of Her2/neu-positive breast cancer cells with Ad3-K/S/Kn increased the killing of cancer cells by the Her2/neu-targeting monoclonal antibody trastuzumab (Herceptin). This study widens our understanding of how Ads achieve high avidity to their receptors and the infection of epithelial tissue. The small recombinant protein Ad3-K/S/Kn has practical implications for the therapy of epithelial cancer and gene/drug delivery to normal epithelial tissues. PMID:21525338

Wang, Hongjie; Li, ZongYi; Yumul, Roma; Lara, Stephanie; Hemminki, Akseli; Fender, Pascal; Lieber, Andre



Production of EV71 vaccine candidates  

PubMed Central

Enterovirus 71 (EV71) is now recognized as an emerging neurotropic virus in Asia and with Coxsackie virus (CV) it is the other major causative agent of hand-foot-mouth diseases (HFMD). Effective medications and/or prophylactic vaccines against HFMD are urgently needed. From a scientific (the feasibility of bioprocess, immunological responses and potency in animal challenge model) and business development (cost of goods) points of view, we in this review address and discuss the pros and cons of different EV71 vaccine candidates that have been produced and evaluated in animal models. Epitope-based synthetic peptide vaccine candidates containing residues 211–225 of VP1 formulated with Freund’s adjuvant (CFA/IFA) elicited low EV71 virus neutralizing antibody responses, but were protective in the suckling mouse challenge model. Among recombinant EV71 subunits (rVP1, rVP2 and rVP3) expressed in E. coli, purified and formulated with CFA/IFA, only VP1 elicited mouse antibody responses with measurable EV71-specific virus neutralization titers. Immunization of mice with either a DNA plasmid containing VP1 gene or VP1 expressed in Salmonella typhimurium also generated neutralizing antibody responses and protected animals against a live EV71 challenge. Recombinant EV71 virus-like particles (rVLP) produced from baculovirus formulated either with CFA/IFA or alum elicited good virus neutralization titers in both mice and non-human primates, and were found to be protective in the suckling mouse EV71 challenge model. Synthetic peptides or recombinant EV71 subunit vaccines (rVP1 and rVLP) formulated in alum were found to be poorly immunogenic in rabbits. Only formalin-inactivated (FI) EV71 virions formulated in alum elicited cross-neutralizing antibodies against different EV71 genotypes in mice, rabbits and non-human primates but induced weak neutralizing responses against CAV16. From a regulatory, economic and market acceptability standpoint, FI-EV71 virion vaccines are the most promising candidates and are currently being evaluated in human clinical trials. We further describe and analyze some new bioprocesses technologies that have great potential applications in EV71 vaccine development. This review also demonstrates the opportunities and challenges that the Asian vaccine industry faces today. PMID:22992566

Chong, Pele; Hsieh, Shih-Yang; Liu, Chia-Chyi; Chou, Ai-Hsiang; Chang, Jui-Yuan; Wu, Suh-Chin; Liu, Shih-Jen; Chow, Yen-Hung; Su, Ih-Jen; Klein, Michel



Enterovirus infections with special reference to enterovirus 71.  


The enteroviruses comprise a large group of immunologically distinct serotypes of viruses belonging to the family of Picornaviridae. Many enteroviruses cause diseases in human, but the infections are generally mild as asymptomatic, therefore, enteroviruses are considered to be unimportant as human pathogens. However, enteroviruses may also result in serious or even fatal disease (as shown in the enterovirus 71 (EV71) epidemic in Taiwan in 1998). There are three types of polioviruses, Coxsackievirus group A and group B viruses, and echoviruses group. All together a total of 67 types are available. Starting from enterovirus type 68 to 71, they are named as enterovirus types. Enterovirus type 72 is hepatitis A virus. Paralytic disease of poliomyelitis was recorded in ancient time but characterization of poliovirus was not reported until the turn of the 19th century that poliomyelitis was a viral disease. The major breakthrough for diagnosing and controlling of poliomyelitis was the discovery that poliovirus can be propagated in human embryonic tissues in cultures. As soon as cultures of human and monkey cells began to use for isolating polioviruses in stool specimen of patients, more unknown viruses were isolated which unlike polioviruses nor Coxsackie viruses; they were called "orphan" viruses or human enteric viruses, name later simplified to "echoviruses". Morphologically all enteroviruses are alike. They are small, ether insensitive viruses with an RNA genome. Their nucleic acid is single stranded, and the nucleocapsid has a cubic (icosahedral) symmetry, and is naked. The host ranges of enteroviruses vary greatly from one type to the next and even among strains of the same type. Polioviruses have a very restricted host range among laboratory animals. Virus isolation is the best method for diagnosis of enterovirus infection, but infection in the central nervous system (CNS) may be detected by polymerase chain reaction (PCR). Currently final identification and serotyping of enteroviruses are by indirect immunofluorescent tests using monoclonal antibody or by neutralization test using antiserum pools described by Lim and Benyesh-Melnick. The incidence and prevalence of diseases associated with the enterovirus infections are varied. The circulation of enteroviruses recently in Tainan and the epidemic of EV71 in Taiwan in 1998 are described in this review. Although poliovirus infection may be eradicated from the world due to the efficient vaccination program, there is no specific antiviral agents for either treatment or prevention for other enterovirus infections. In 1991, a new antiviral "pleconaril" which is a novel orally bioavailable and systematically acting small molecule inhibitor for picornaviruses. "Pleconaril" is currently in clinical trials for treatment of enterovirus meningitis and respiratory infections. PMID:10806956

Hsiung, G D; Wang, J R



Detection of viral bioagents using a shear horizontal surface acoustic wave biosensor.  


Viruses are of high medical and biodefense concern and their detection at concentrations well below the threshold necessary to cause health hazards continues to be a challenge with respect to sensitivity, specificity, and selectivity. Ideally, assays for accurate and real time detection of viral agents would not necessitate any pre-processing of the analyte, which would make them applicable for example to bodily fluids (blood, sputum) and man-made as well as naturally occurring bodies of water (pools, rivers). We describe herein a robust biosensor that combines the sensitivity of surface acoustic waves (SAW) generated at a frequency of 325MHz with the specificity provided by antibodies for the detection of viral agents. A lithium tantalate-based SAW transducer with silicon dioxide waveguide sensor platform featuring three test and one reference delay lines was used to adsorb antibodies directed against either Coxsackie virus B4 or the category A bioagent Sin Nombre virus (SNV), a member of the genus Hantavirus, family Bunyaviridae, negative-stranded RNA viruses. Rapid detection (within seconds) of increasing concentrations of viral particles was linear over a range of order of magnitude for both viruses, although the sensor was approximately 5 x 10(5)-fold more sensitive for the detection of SNV. For both pathogens, the sensor's selectivity for its target was not compromised by the presence of confounding Herpes Simplex virus type 1. The biosensor was able to detect SNV at doses lower than the load of virus typically found in a human patient suffering from hantavirus cardiopulmonary syndrome (HCPS). Further, in a proof-of-principle real world application, the SAW biosensor was capable to selectively detect SNV agents in complex solutions, such as naturally occurring bodies of water (river, sewage effluent) without analyte pre-processing. This is the first study that reports on the detection of viral agents using an antibody-based SAW biosensor that has the potential to be used as a hand-held and self-contained device for rapid viral detection in the field. PMID:18262781

Bisoffi, M; Hjelle, B; Brown, D C; Branch, D W; Edwards, T L; Brozik, S M; Bondu-Hawkins, V S; Larson, R S



Probing the structure and function of biopolymer-carbon nanotube hybrids with molecular dynamics  

NASA Astrophysics Data System (ADS)

Nanoscience deals with the characterization and manipulation of matter on the atomic/molecular size scale in order to deepen our understanding of condensed matter and develop revolutionary technology. Meeting the demands of the rapidly advancing nanotechnological frontier requires novel, multifunctional nanoscale materials. Among the most promising nanomaterials to fulfill this need are biopolymer-carbon nanotube hybrids (Bio-CNT). Bio-CNT consists of a single-walled carbon nanotube (CNT) coated with a self-assembled layer of biopolymers such as DNA or protein. Experiments have demonstrated that these nanomaterials possess a wide range of technologically useful properties with applications in nanoelectronics, medicine, homeland security, environmental safety and microbiology. However, a fundamental understanding of the self-assembly mechanics, structure and energetics of Bio-CNT is lacking. The objective of this thesis is to address this deficiency through molecular dynamics (MD) simulation, which provides an atomic-scale window into the behavior of this unique nanomaterial. MD shows that Bio-CNT composed of single-stranded DNA (ssDNA) self-assembles via the formation of high affinity contacts between DNA bases and the CNT sidewall. Calculation of the base-CNT binding free energy by thermodynamic integration reveals that these contacts result from the attractive pi--pi stacking interaction. Binding affinities follow the trend G > A > T > C. MD reveals that long ssDNA sequences are driven into a helical wrapping about CNT with a sub-10 nm pitch by electrostatic and torsional interactions in the backbone. A large-scale replica exchange molecular dynamics simulation reveals that ssDNA-CNT hybrids are disordered. At room temperature, ssDNA can reside in several low-energy conformations that contain a sequence-specific arrangement of bases detached from CNT surface. MD demonstrates that protein-CNT hybrids composed of the Coxsackie-adenovirus receptor are biologically active and function as a nanobiosensor with specific recognition of Knob proteins from the adenovirus capsid. Simulation also shows that the rigid CNT damps structural fluctuations in bound proteins, which may have important ramifications for biosensing devices composed of protein-CNT hybrids. These results expand current knowledge of Bio-CNT and demonstrate the effectiveness of MD for investigations of nanobiomolecular systems.

Johnson, Robert R.


Hand, Foot and Mouth Disease in China: Patterns of Spread and Transmissibility during 2008-2009  

PubMed Central

Background Large outbreaks of hand, foot and mouth disease (HFMD) were observed in both 2008 and 2009 in China. Methods Using the national surveillance data since May 2, 2008, epidemiological characteristics of the outbreaks are summarized, and the transmissibility of the disease and the effects of potential risk factors were evaluated via a susceptible-infectious-recovered transmission model. Results Children of 1.0–2.9 years were the most susceptible group to HFMD (odds ratios [OR] > 2.3 as compared to other age groups). Infant cases had the highest incidences of severe disease (ORs > 1.4) and death (ORs > 2.4), as well as the longest delay from symptom onset to diagnosis (2.3 days). Males were more susceptible to HFMD than females (OR=1.56 [95% confidence interval=1.56, 1.57]). An one day delay in diagnosis was associated with increases in the odds of severe disease by 40.3% [38.7%, 41.9%] and in the odds of death by 53.7% [43.6%, 64.5%]. Compared to Coxsackie A16, enterovirus (EV) 71 is more strongly associated with severe disease (OR=15.6 [13.4, 18.1]) and death (OR=40.7 [13.0, 127.3]). The estimated local effective reproductive numbers among prefectures ranged from 1.4 to 1.6 (median=1.4) in spring and stayed below 1.2 in other seasons. A higher risk of transmission was associated with temperatures in the range of 70-80F, higher relative humidity, wind speed, precipitation, population density, and the periods in which schools were open. Conclusion HFMD is a moderately transmittable infectious disease, mainly among pre-school children. EV71 was responsible for most severe cases and fatalities. Mixing of asymptomatically infected children in schools might have contributed to the spread of HFMD. Timely diagnosis may be a key to reducing the high mortality rate in infants. PMID:21968769

Wang, Yu; Feng, Zijian; Yang, Yang; Self, Steve; Gao, Yongjun; Longini, Ira M.; Wakefield, Jon; Zhang, Jing; Wang, Liping; Chen, Xi; Yao, Lena; Stanaway, Jeffrey D.; Wang, Zijun; Yang, Weizhong



Treatment of Radioresistant Stem-Like Esophageal Cancer Cells by an Apoptotic Gene-Armed, Telomerase-Specific Oncolytic Adenovirus  

PubMed Central

Purpose Radioresistance may be caused by cancer stem cells (CSCs). Because CSCs require telomerase to proliferate, a telomerase-specific oncolytic adenoviral vector carrying apoptotic TRAIL and E1A gene (Ad/TRAIL-E1) may preferentially target CSCs. Experimental Design We established two pairs of parental and radioresistant (R) esophageal carcinoma cell lines (Seg-1, Seg-1R and TE-2, TE-2R) by fractionated irradiation (FIR). Stem cell markers were measured by Western blotting and flow cytometry. Serial sorting was used to enrich stem-like side population (SP) cells. Telomerase activity, transgene expression, antitumor activity, apoptosis induction, and viral replication were determined in vitro and/or in vivo. Results Expression of the stem cell markers ?-catenin, Oct3/4, and ?1-integrin in Seg-1R cells was 29.4%, 27.5%, and 97.3%, respectively, compared with 4.8%, 14.9%, and 45.3% in Seg-1 cells (P < 0.05). SP levels in Seg-1R and TE-2R cells were 14.6% and 2.7%, respectively, compared with 3.4% and 0.3% in Seg-1 and TE-2 cells. Serial sorting of Seg-1R SP cells demonstrated enrichment of the SP cells. Telomerase activity in Seg-1R, Seg-1R SP and TE-2R cells were significantly higher than Seg-1, Seg-1R non-SP and TE-2 cells respectively (P < 0.05). Seg-1R and TE-2R cells were more sensitive to Ad/TRAIL-E1 than were parental cells. Increased Coxsackie-adenovirus receptor (CAR) and elevated transgene expressions were found in the radioresistant cells. Ad/TRAIL-E1 resulted in significant tumor growth suppression and longer survival in Seg-1R-bearing mice (P < 0.05) with no significant toxicity. Conclusion Radioresistant cells established by FIR display CSC-like cell properties. Ad/TRAIL-E1 preferentially targets radioresistant CSC-like cells. PMID:18451249

Zhang, Xiaochun; Komaki, Ritsuko; Wang, Li; Fang, Bingliang; Chang, Joe Y.



Serologic characterization and sero-epidemiologic studies on acute hemorrhagic conjunctivitis (AHC) virus.  


Serologic and sero-epidemiologic characteristics of AHC virus infection were studied by neutralization test (NT). Four-fold or greater virus neutralizing (VN) antibody response was demonstrated to the Japanese isolate of AHC virus (the J 670/71 strain) in 77.3% and 66.7% of paired sera from clinical AHC patients in Japan (1971-1973) and Tunisia (1973). The four patients from Indonesia studied in 1972 showed similar antibody response. Cross-neutralization tests of AHC virus isolated in Japan (1971), Taiwan (1971), Hong Kong (1971), Thailand (1972), Indonesia (1972), Singapore (1972), Morocco (1971) and England (1971) with three kinds of antisera prepared against Japanese, Hong Kong and Moroccan AHC virus isolates indicated their antigenic identity. However, isolates from Sinapore in 1970 (Singapore 70 virus) were not neutralized with the AHC virus antisera mentioned above: Singapore 70 virus constitutes another antigenic type, to which, however, no VN antibody rise was found in paired patients' sera from Japan, Tunisia and Indonesia. Thus, no serologic evidence supporting an etiologic role of this virus group in the development of AHC was found. Although cross-tests using monospecific antisera suggested some cross-relation between AHC and both echovirus type 4 (E4) and coxsackie A (CA), type 19, no serologic relationship between AHC and these viruses was found. Sera from healthy individuals collected before and after AHC outbreaks were tested for VN antibody against AHC virus in Japan and two epidemic foci, Ghana and Indonesia. Before the epidemic, 80 to 90% of the people lacked antibody in the three countries, but 39.7% and 45.2% of inhabitants posessed VN antibody of 1:8 or over in Ghana and Indonesia after the outbreak. In Japan, however, only a slight increase was found in VN antibody prevalence afterwards. Serologic study showed that 41.5% of horse sera were VN positive at dilutions of 1:8 or more; many cattle sera also had a low VN titer but few cynomologus monkey sera had VN activity. PMID:165719

Kono, R; Sasagawa, A; Miyamura, K; Tajiri, E



Formalin-Inactivated EV71 Vaccine Candidate Induced Cross-Neutralizing Antibody against Subgenotypes B1, B4, B5 and C4A in Adult Volunteers  

PubMed Central

Background Enterovirus 71 (EV71) has caused several epidemics of hand, foot and mouth diseases (HFMD) in Asia. No effective EV71 vaccine is available. A randomized and open-label phase I clinical study registered with #NCT01268787, aims to evaluate the safety, reactogenicity and immunogenicity of a formalin-inactivated EV71 vaccine candidate (EV71vac) at 5- and 10-µg doses. In this study we report the cross-neutralizing antibody responses from each volunteer against different subgenotypes of EV71 and CVA16. Methods Sixty eligible healthy adults were recruited and vaccinated. Blood samples were obtained on day 0, 21 and 42 and tested against B1, B4, B5, C2, C4A, C4B and CVA16 for cross-neutralizing antibody responses. Results The immunogenicity of both 5- and 10- µg doses were found to be very similar. Approximately 45% of the participants had <8 pre-vaccination neutralization titers (Nt) against the B4 vaccine strain. After the first EV71vac immunization, 95% of vaccinees have >4-fold increase in Nt, but there was no further increase in Nt after the second dose. EV71vac induced very strong cross-neutralizing antibody responses in >85% of volunteers without pre-existing Nt against subgenotype B1, B5 and C4A. EV71vac elicited weak cross-neutralizing antibody responses (?20% of participants) against a C4B and Coxsackie virus A16. Over 90% of vaccinated volunteers did not develop cross-neutralizing antibody responses (Nt<8) against a C2 strain. EV71vac can boost and significantly enhance the neutralizing antibody responses in volunteers who already had pre-vaccination antibodies against EV71 and/or CVA16. Conclusion EV71vac is efficient in eliciting cross-neutralizing antibody responses against EV71 subgenotypes B1, B4, B5, and C4A, and provides the rationale for its evaluation in phase II clinical trials. Trial Registration __NCT01268787 PMID:24278177

Chang, Jui-Yuan; Jiang, Renee; Hsieh, Yi-Chin; Tsao, Amanda; Wu, Chien-Long; Huang, Ju-Lan; Fung, Chang-Phone; Hsieh, Szu-Min; Wang, Ya-Fang; Wang, Jen-Ren; Hu, Mei-Hua; Chiang, Jen-Ron; Su, Ih-Jen; Chong, Pele Choi-Sing