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1

Molecular typing and characterization of a new serotype of human enterovirus (EV-B111) identified in China.  

PubMed

Molecular methods, based on sequencing the region encoding the complete VP1 or P1 protein, have enabled the rapid identification of new enterovirus serotypes. In the present study, the complete genome of a newly discovered enterovirus serotype, strain Q0011/XZ/CHN/2000 (hereafter referred to as Q0011), was sequenced and analyzed. The virus, isolated from a stool sample from a patient with acute flaccid paralysis in the Tibet region of China in 2000, was characterized by amplicon sequencing and comparison to a GenBank database of enterovirus nucleotide sequences. The nucleotide sequence encoding the complete VP1 capsid protein is most closely related to the sequences of viruses within the species enterovirus B (EV-B), but is less than 72.1% identical to the homologous sequences of the recognized human enterovirus serotypes, with the greatest homology to EV-B101 and echovirus 32. Moreover, the deduced amino acid sequence of the complete VP1 region is less than 84.7% identical to those of the recognized serotypes, suggesting that the strain is a new serotype of enterovirus within EV-B. The virus was characterized as a new enterovirus type, named EV-B111, by the Picornaviridae Study Group of the International Committee on Taxonomy of Viruses. Low positive rate and titer of neutralizing antibody against EV-B111 were found in the Tibet region of China. Nearly 50% of children ?5 years had no neutralizing antibody against EV-B111. So the extent of transmission and the exposure of the population to this new EV are very limited. This is the first identification of a new serotype of human enterovirus in China, and strain Q0011 was designated the prototype strain of EV-B111. PMID:24503225

Zhang, Yong; Hong, Mei; Sun, Qiang; Zhu, Shuangli; Tsewang; Li, Xiaolei; Yan, Dongmei; Wang, Dongyan; Xu, Wenbo

2014-04-01

2

Human enterovirus 71 subgenotype B3 lacks coxsackievirus A16-like neurovirulence in mice infection  

PubMed Central

Background At least three different EV-71 subgenotypes were identified from an outbreak in Malaysia in 1998. The subgenotypes C2 and B4 were associated with the severe and fatal infections, whereas the B3 virus was associated with mild to subclinical infections. The B3 virus genome sequences had ?85% similarity at the 3' end to CV-A16. This offers opportunities to examine if there are characteristic similarities and differences in virulence between CV-A16, EV-71 B3 and EV-71 B4 and to determine if the presence of the CV-A16-liked genes in EV-71 B3 would also confer the virus with a CV-A16-liked neurovirulence in mice model infection. Results Analysis of human enterovirus 71 (EV-71) subgenotype B3 genome sequences revealed that the 3D RNA polymerase and domain Z of the 3'-untranslating region RNA secondary structure had high similarity to CV-A16. Intracerebral inoculation of one-day old mice with the virus resulted in 16% of the mice showing swollen hind limbs and significantly lower weight gain in comparison to EV-71 B4-infected mice. None of the mice presented with hind leg paralysis typical in all the CV-A16 infected mice. CV-A16 genome sequences were amplified from the CV-A16-infected mice brain but no amplification was obtained from all the EV-71-inoculated mice suggesting that no replication had taken place in the suckling mice brain. Conclusion The findings presented here suggest that EV-71 B3 viruses had CV-A16-liked non-structural gene features at the 3'-end of the genome. Their presence could have affected virulence by affecting the mice general health but was insufficient to confer the EV-71 B3 virus a CV-A16-liked neurovirulence in mice model infection. PMID:16122396

Chan, Yoke-Fun; AbuBakar, Sazaly

2005-01-01

3

Genetic Relationship between Cocirculating Human Enteroviruses Species C  

E-print Network

Genetic Relationship between Cocirculating Human Enteroviruses Species C Mae¨l Bessaud1 Recombination events between human enteroviruses (HEV) are known to occur frequently and to participate to the Human enterovirus species C (HEV-C) that had been cocirculating in a small geographic area of Madagascar

Paris-Sud XI, Université de

4

Accuracy of Diagnostic Methods and Surveillance Sensitivity for Human Enterovirus, South Korea, 1999-2011  

PubMed Central

The epidemiology of enteroviral infection in South Korea during 19992011 chronicles nationwide outbreaks and changing detection and subtyping methods used over the 13-year period. Of 14,657 patients whose samples were tested, 4,762 (32.5%) samples were positive for human enterovirus (human EV); as diagnostic methods improved, the rate of positive results increased. A seasonal trend of outbreaks was documented. Genotypes enterovirus 71, echovirus 30, coxsackievirus B5, enterovirus 6, and coxsackievirus B2 were the most common genotypes identified. Accurate test results correlated clinical syndromes to enterovirus genotypes: aseptic meningitis to echovirus 30, enterovirus 6, and coxsackievirus B5; hand, foot and mouth disease to coxsackievirus A16; and hand, foot and mouth disease with neurologic complications to enterovirus 71. There are currently no treatments specific to human EV infections; surveillance of enterovirus infections such as this study provides may assist with evaluating the need to research and develop treatments for infections caused by virulent human EV genotypes. PMID:23876671

Hyeon, Ji-Yeon; Hwang, Seoyeon; Kim, Hyejin; Song, Jaehyoung; Ahn, Jeongbae; Kang, Byunghak; Kim, Kisoon; Choi, Wooyoung; Chung, Jae Keun; Kim, Cheon-Hyun; Cho, Kyungsoon; Jee, Youngmee; Kim, Jonghyun; Kim, Kisang; Kim, Sun-Hee; Kim, Min-Ji

2013-01-01

5

Epidemiology and seroepidemiology of human enterovirus 71 among Thai populations  

PubMed Central

Background Human enterovirus 71 (EV71) is an important pathogen caused large outbreaks in Asian-Pacific region with severe neurological complications and may lead to death in young children. Understanding of the etiological spectrum and epidemic changes of enterovirus and populations immunity against EV71 are crucial for the implementation of future therapeutic and prophylactic intervention. Results A total of 1,182 patients who presented with the symptoms of hand foot and mouth disease (67.3%) or herpangina (HA) (16.7%) and admitted to the hospitals during 2008-2013 were tested for enterovirus using pan-enterovirus PCR targeting 5?-untranslated region and specific PCR for viral capsid protein 1 gene. Overall, 59.7% were pan-enterovirus positive comprising 9.1% EV71 and 31.2% coxsackievirus species A (CV-A) including 70.5% CV-A6, 27.6% CV-A16, 1.1% CV-A10, and 0.8% CV-A5. HFMD and HA occurred endemically during 2008-2011. The number of cases increased dramatically in June 2012 with the percentage of the recently emerged CV-A6 significantly rose to 28.4%. Co-circulation between different EV71 genotypes was observed during the outbreak. Total of 161 sera obtained from healthy individuals were tested for neutralizing antibodies (NAb) against EV71 subgenotype B5 (EV71-B5) using microneutralization assay. The seropositive rate of EV71-B5 was 65.8%. The age-adjusted seroprevalence for individuals was found to be lowest in children aged >6months to 2years (42.5%). The seropositive rate remained relatively low in preschool children aged?>?2years to 6years (48.3%) and thereafter increased sharply to more than 80% in individuals aged?>?6years. Conclusions This study describes longitudinal data reflecting changing patterns of enterovirus prevalence over 6years and demonstrates high seroprevalences of EV71-B5 NAb among Thai individuals. The rate of EV71 seropositive increased with age but without gender-specific significant difference. We identified that relative lower EV71 seropositive rate in early 2012 may demonstrate widely presented of EV71-B5 in the population before account for a large outbreak scale epidemic occurred in 2012 with due to a relatively high susceptibility of the younger population. PMID:24548776

2014-01-01

6

[Research progress in human enterovirus recombination].  

PubMed

Complex genetic variation has been known to occur during the transmission of human enterovirus (HEV), and the HEV virulence and pathogenicity enhanced by genetic recombination also pose a serious threat to human health. In recent years, the interest in recombination mechanism of genetic plasticity has been renewed with the emergence of pathogenic recombinant circulating vaccine-derived polioviruses, which were implicated in poliomyelitis outbreaks in several regions of the world with insufficient vaccination coverage. This paper reviews recent research progress in HEV genome, including evolutionary characteristics, recombination types, and in vitro recombinant construction. PMID:24923173

Wang, Hai-Yan; Jiang, Ping; Chen, Peng; Tao, Ze-Xin; Zhang, Wen-Qiang; Xu, Ai-Qiang

2014-03-01

7

Crystal Structure of Human Enterovirus 71  

SciTech Connect

Enterovirus 71 is a picornavirus associated with fatal neurological illness in infants and young children. Here, we report the crystal structure of enterovirus 71 and show that, unlike in other enteroviruses, the 'pocket factor,' a small molecule that stabilizes the virus, is partly exposed on the floor of the 'canyon.' Thus, the structure of antiviral compounds may require a hydrophilic head group designed to interact with residues at the entrance of the pocket.

Plevka, Pavel; Perera, Rushika; Cardosa, Jane; Kuhn, Richard J.; Rossmann, Michael G. (Purdue); (Sentinext)

2013-04-08

8

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

2013-01-01

9

Characterization of a Novel Enterovirus Serotype and an Enterovirus EV-B93 Isolated from Acute Flaccid Paralysis Patients  

PubMed Central

Non-polio enteroviruses (NPEVs) are among the most common viruses infecting humans worldwide. Most of these infections are asymptomatic but few can lead to systemic and neurological disorders like Acute Flaccid Paralysis (AFP). Acute Flaccid Paralysis is a clinical syndrome and NPEVs have been isolated frequently from the patients suffering from AFP but little is known about their causal relationship. The objective of this study was to identify and characterize the NPEV serotypes recovered from 184 stool samples collected from AFP patients in Federally Administered Tribal Areas (FATA) in north-west of Pakistan. Overall, 44 (95.6 %) isolates were successfully typed through microneutralization assay as a member of enterovirus B species including echovirus (E)-2, E-3, E-4, E-6, E-7, E-11, E-13, E-14, E-21 and E-29 while two isolates (PAK NIH SP6545B and PAK NIH SP1202B) remained untypeable. The VP1 and capsid regions analysis characterized these viruses as EV-B93 and EV-B106. Phylogenetic analysis confirmed that PAK NIH isolates had high genetic diversity and represent distinct genotypes circulating in the country. Our findings highlight the role of NPEVs in AFP cases to be thoroughly investigated especially in high disease risk areas, with limited surveillance activities and health resources. PMID:24244603

Shaukat, Shahzad; Angez, Mehar; Alam, Muhammad Masroor; Sharif, Salmaan; Khurshid, Adnan; Mahmood, Tariq; Zaidi, Syed Sohail Zahoor

2013-01-01

10

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

PubMed

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

2008-08-15

11

Enterovirus infections with special reference to enterovirus 71.  

PubMed

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

2000-03-01

12

Deferoxamine compensates for decreases in B cell counts and reduces mortality in enterovirus 71-infected mice.  

PubMed

Enterovirus 71 is one of the major causative agents of hand, foot and mouth disease in children under six years of age. No vaccine or antiviral therapy is currently available. In this work, we found that the number of B cells was reduced in enterovirus 71-infected mice. Deferoxamine, a marine microbial natural product, compensated for the decreased levels of B cells caused by enterovirus 71 infection. The neutralizing antibody titer was also improved after deferoxamine treatment. Furthermore, deferoxamine relieved symptoms and reduced mortality and muscle damage caused by enterovirus 71 infection. This work suggested that deferoxamine has the potential for further development as a B cell-immunomodulator against enterovirus 71. PMID:25003792

Yang, Yajun; Ma, Jing; Xiu, Jinghui; Bai, Lin; Guan, Feifei; Zhang, Li; Liu, Jiangning; Zhang, Lianfeng

2014-07-01

13

Complete Genome Characterization of a Novel Enterovirus Type EV-B106 Isolated in China, 2012  

PubMed Central

Human enterovirus B106 (EV-B106) is a recently identified member of enterovirus species B. In this study, we report the complete genomic characterization of an EV-B106 strain (148/YN/CHN/12) isolated from an acute flaccid paralysis patient in Yunnan Province, China. The new strain had 79.281.3% nucleotide and 89.194.8% amino acid similarity in the VP1 region with the other two EV-B106 strains from Bolivia and Pakistan. When compared with other EV serotypes, it had the highest (73.3%) VP1 nucleotide similarity with the EV-B77 prototype strain CF496-99. However, when aligned with all EV-B106 and EV-B77 sequences available from the GenBank database, two major frame shifts were observed in the VP1 coding region, which resulted in substantial (20.5%) VP1 amino acid divergence between the two serotypes. Phylogenetic analysis and similarity plot analysis revealed multiple recombination events in the genome of this strain. This is the first report of the complete genome of EV-B106. PMID:24584702

Tang, Jingjing; Tao, Zexin; Ding, Zhengrong; Zhang, Yong; Zhang, Jie; Tian, Bingjun; Zhao, Zhixian; Zhang, Lifen; Xu, Wenbo

2014-01-01

14

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

PubMed

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

2001-01-01

15

First Full Genome Sequence of a Human Enterovirus A120, Isolated in Madagascar  

PubMed Central

We report the first complete genome sequence of an enterovirus isolate belonging to the human enterovirus A species of the Picornaviridae family and to type A120 (EV-A120). The EV-A120 isolate MAD-2741-11 was obtained from the stool of a healthy child living on Madagascar Island. The isolate genome was amplified by a reverse transcription-PCR method, and the consensus sequence was determined. PMID:24948760

Joffret, Marie-Line; Delpeyroux, Francis; Heraud, Jean-Michel

2014-01-01

16

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 , 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

2011-01-01

17

Characterization of the genome of human enteroviruses: design of generic primers for amplification and sequencing of different regions of the viral genome  

E-print Network

1 Characterization of the genome of human enteroviruses: design of generic primers.01.027 #12;2 Abstract Human enteroviruses are among the most common viruses infecting humans and can cause. Biodiversity and evolution of human enterovirus genomes are shaped by frequent recombination events. Therefore

Boyer, Edmond

18

Molecular identification and analysis of nonserotypeable human enteroviruses.  

PubMed

Conventional approaches to characterizing human enteroviruses (HEVs) are based on viral isolation and neutralization. Molecular typing methods depend largely on reverse transcription-PCR (RT-PCR) and nucleotide sequencing of the entire or partial VP1 gene. A modified RT-PCR-based reverse line blot (RLB) hybridization assay was developed as a rapid and efficient way to characterize common and nonserotypeable (by neutralization) HEVs. Twenty HEV serotypes accounted for 87.1% of all HEVs isolated at a reference laboratory from 1979 to 2007; these common serotypes were identified using one sense and three antisense primers and a set of 80 serotype-specific probes in VP1 (F. Zhou et al., J. Clin. Microbiol. 47:2737-2743, 2009). In this study, one HEV-specific primer pair, two probes in the 5' untranslated region (UTR), and a new set of 80 serotype-specific probes in VP1 were designed. First, we successfully applied the modified RT-PCR-RLB (using two HEV-specific probes and two sets of serotype-specific probes) to synchronously detect the 5' UTR and VP1 regions of 131/132 isolates previously studied (F. Zhou et al., J. Clin. Microbiol. 47:2737-2743, 2009). Then, this method was used to identify 73/92 nonserotypeable HEV isolates; 19 nonserotypeable isolates were hybridized only with HEV-specific probes. The VP1 region of 92 nonserotypeable HEV isolates was sequenced; 73 sequences corresponded with one or both RLB results and 19 (not belonging to the 20 most common genotypes) were identified only by sequencing. Two sets of serotype-specific probes can capture the majority of strains belonging to the 20 most common serotypes/genotypes simultaneously or complementarily. Synchronous detection of the 5' UTR and VP1 region by RT-PCR-RLB will facilitate the identification of HEVs, especially nonserotypeable isolates. PMID:20164278

Zhou, Fei; Kong, Fanrong; McPhie, Kenneth; Ratnamohan, Mala; Gilbert, Gwendolyn L; Dwyer, Dominic E

2010-04-01

19

Enteroviruses and the pathogenesis of type 1 diabetes revisited: cross-reactivity of enterovirus capsid protein (VP1) antibodies with human mitochondrial proteins.  

PubMed

Current or recent enteroviral infections show an association with type 1 diabetes. However, evidence for this has mainly been generated using a particular mouse monoclonal antibody (clone 5-D8/1) which binds the viral capsid protein VP1. Difficulty in confirming these findings using other independent methods has led to the concern that this might be artefactual. To address this, we examined the potential cross-reactivity of clone 5-D8/1 with normal islet proteins. Western blotting, two-dimensional gel electrophoresis, and mass spectrometry were used to identify human islet proteins bound by the clone 5-D8/1. We found a distinct reactivity with two mitochondrial proteins, creatine kinase B-type and ATP synthase beta subunit. Immunohistochemistry using the clone 5-D8/1 revealed a granular cytoplasmic staining pattern in mitochondria-rich cells, ie hepatocytes, ductal epithelial cells, vascular endothelial cells, skeletal muscle cells, and the neoplastic salivary gland oncocytoma cells, whereas connective tissue and infiltrating immune cells were negative. Staining on islets of Langerhans from subjects with recent-onset type 1 diabetes, but not on isolated human islets infected in vitro with enteroviruses, could be blocked after mixing the clone 5-D8/1 with the mitochondrial proteins. Collectively, our data show that the clone 5-D8/1 detects two human mitochondrial enzymes in addition to enteroviral VP1. The notion that the previously reported VP1 positivity in islets of recent-onset type 1 diabetes patients could reflect cross-reactivity to native islet proteins and not the presence of EV is supported by difficulties in demonstrating EV infection by independent techniques such as PCR or in situ hybridization. These findings call for revisiting the presence of enteroviruses in pancreatic islets of patients with type 1 diabetes. PMID:23335350

Hansson, Sara F; Korsgren, Stella; Pontn, Fredrik; Korsgren, Olle

2013-04-01

20

Survey of human enterovirus occurrence in fresh and marine surface waters on Long Island.  

PubMed Central

A variety of surface water systems, including a lake, a creek, and two marine embayments, were analyzed on a monthly basis for indigenous human enteroviruses and coliform bacteria. Findings are discussed in terms of the probable pollution sources to each system and their relationship to data from previous studies. PMID:229767

Vaughn, J M; Landry, E F; Thomas, M Z; Vicale, T J; Penello, W F

1979-01-01

21

Estimation of Contamination Sources of Human Enteroviruses in a Wastewater Treatment and Reclamation System by PCR-DGGE.  

PubMed

A polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) method was employed to estimate the contamination sources of human enteroviruses and understand how their dominant strains vary in a wastewater treatment and reclamation system consisting of sewage collection, wastewater treatment with membrane bioreactor and open lakes for reclaimed water storage and reuse. After PCR-DGGE using a selected primer set targeting enteroviruses, phylogenetic analysis of acquired enterovirus gene sequences was performed. Enteroviruses identified from the septic tank were much more diverse than those from grey water and kitchen wastewater. Several unique types of enterovirus different from those in wastewater samples were dominant in a biological wastewater treatment unit. Membrane filtration followed by chlorination was proved effective for physically eliminating enteroviruses; however, secondary contamination likely occurred as the reclaimed water was stored in artificial lakes. Enterovirus 71 (EV71), a hand-foot-and-mouth disease (HFMD) viral pathogen, was detected mainly from the artificial lakes, implying that wastewater effluent was not the contamination source of EV71 and that there were unidentified non-point sources of the contamination with the HFMD viral pathogen in the reclaimed water stored in the artificial lakes. The PCR-DGGE targeting enteroviruses provided robust evidence about viral contamination sources in the wastewater treatment and reclamation system. PMID:24715657

Ji, Zheng; Wang, Xiaochang C; Xu, Limei; Zhang, Chongmiao; Funamizu, Naoyuki; Okabe, Satoshi; Sano, Daisuke

2014-06-01

22

Use of the 5' untranslated region and VP1 region to examine the molecular diversity in enterovirus B species.  

PubMed

Human enteroviruses evolve quickly. The 5' untranslated region (UTR) is fundamentally important for efficient viral replication and for virulence; the VP1 region correlates well with antigenic typing by neutralization, and can be used for virus identification and evolutionary studies. In order to investigate the molecular diversity in EV-B species, the 5' UTR and VP1 regions were analysed for 208 clinical isolates from a single public-health laboratory (serving New South Wales, Australia), representing 28 EV-B types. Sequences were compared with the 5' UTR and VP1 regions of 98 strains available in GenBank, representing the same 28 types. The genetic relationships were analysed using two types of software (mega and BioNumerics). The sequence analyses of the 5' UTR and VP1 regions of 306 EV-B strains demonstrated that: (i) comparing the two regions gives strong evidence of epidemiological linkage of strains in some serotypes; (ii) the intraserotypic genetic variation within each gene reveals that they evolve distinctly largely due to their different functions; and (iii) mutation and possible recombination in the two regions play significant roles in the molecular diversity of EV-B. Understanding the tempo and pattern of molecular diversity and evolution is of great importance in the pathogenesis of EV-B enteroviruses, information which will assist in disease prevention and control. PMID:25038138

Zhou, Fei; Wang, Qinning; Sintchenko, Vitali; Gilbert, Gwendolyn L; O'Sullivan, Matthew V N; Iredell, Jonathan R; Dwyer, Dominic E

2014-10-01

23

Molecular detection and characterization of human enteroviruses in Korean surface water.  

PubMed

In this study, the genetic epidemiology of enteroviruses (EVs) in Korean surface water was evaluated by conducting phylogenetic analyses of the nucleotide sequences of the 5' non-coding region (5' NCR), which was determined by RT-PCR analysis of total culturable virus assay-positive samples. The results showed that the nucleotide sequences of the EVs could be classified into 4 genetic clusters, and that the predominant presence of Korea EVs were very similar to echoviruses type 30. Interestingly, two nucleotide sequences were very similar to those of coxsackievirus type B1 isolated from aseptic meningitis patients in Seoul, Korea, implying the possibility of a common source for the viruses circulated in water systems and humans. In addition, 3 nucleotide sequences clustered strongly with the nucleotide sequences from China or Japan, and one fell into the same cluster as echovirus type 11 from Taiwan, which suggests that EVs in Asia may have evolved in a region-specific manner. Taken together, the results of this study revealed that EVs from Korea surface waters could be genetically classified as coxsackieviruses or echoviruses, and that they evolved in Asia in a region-specific manner. PMID:18604502

Lee, Gyucheol; Lee, Chanhee

2008-06-01

24

Outbreak of herpangina in the Brazilian Amazon in 2009 caused by Enterovirus B.  

PubMed

In October 2009, our laboratory was contacted by a Brazilian Public Health organization regarding a severe community outbreak of an acute exanthematic and febrile disease in the Brazilian Amazon that primarily affected children. A total of 44 patients with febrile disease were identified by the local public health system, 37 of whom were children between 1 and 9 years of age. Molecular virological and phylogenetic characterization revealed that enterovirus B was the etiological agent of this outbreak, which was characterized by a clinical presentation known as herpangina. PMID:24197788

Oliveira, D B; Campos, R K; Soares, M S; Barros, R B; Batista, T C A; Ferreira, P C P; Bonjardim, C A; Trindade, G S; Abraho, J S; Kroon, Erna Geessien

2014-05-01

25

Attenuation of Human Enterovirus 71 High-Replication-Fidelity Variants in AG129 Mice  

PubMed Central

ABSTRACT In a screen for ribavirin resistance, a novel high-fidelity variant of human enterovirus 71 (EV71) with the single amino acid change L123F in its RNA-dependent RNA polymerase (RdRp or 3D) was identified. Based on the crystal structure of EV71 RdRp, L123 locates at the entrance of the RNA template binding channel, which might form a fidelity checkpoint. EV71 RdRp-L123F variants generated less progeny in a guanidine resistance assay and virus populations with lower mutation frequencies in cell culture passage due to their higher replication fidelity. However, compared with wild-type viruses, they did not show growth defects. In vivo infections further revealed that high-fidelity mutations L123F and G64R (previously reported) negatively impacted EV71 fitness and greatly reduced viral pathogenicity alone or together in AG129 mice. Interestingly, a variant with double mutations, RG/B4-G64R/L123F (where RG/B4 is an EV71 genotype B4 virus constructed by reverse genetics [RG])showed higher fidelity in vitro and less virulence in vivo than any one of the above two single mutants. The 50% lethal dose (LD50) of the double mutant increased more than 500 times compared with the LD50 of wild-type RG/B4 in mice. The results indicated that these high-fidelity variants exhibited an attenuated pathogenic phenotype in vivo and offer promise as a live attenuated EV71 vaccine. IMPORTANCE The error-prone nature of the RNA-dependent RNA polymerase (RdRp) of RNA viruses during replication results in quasispecies and aids survival of virus populations under a wide range of selective pressures. Virus variants with higher replication fidelity exhibit lower genetic diversity and attenuated pathogenicity in vivo. Here, we identified a novel high-fidelity mutation L123F in the RdRp of human enterovirus 71 (EV71). We further elucidated that EV71 variants with the RdRp-L123F mutation and/or the previously identified high-fidelity mutation RdRp-G64R were attenuated in an AG129 mouse model. As EV71 has emerged as a serious worldwide health threat, especially in developing countries in the Asia-Pacific region, we urgently need EV71 vaccines. Learning from the poliovirus vaccination, we prefer live attenuated EV71 vaccines to inactivated EV71 vaccines in order to effectively control EV71 outbreaks at low cost. Our results imply a new means of attenuating EV71 and reducing its mutation rate at the same time. PMID:24623423

Meng, Tao

2014-01-01

26

A neonatal gnotobiotic pig model of human enterovirus 71 infection and associated immune responses  

PubMed Central

Vaccine development and pathogenesis studies for human enterovirus 71 are limited by a lack of suitable animal models. Here, we report the development of a novel neonatal gnotobiotic pig model using the non-pig-adapted neurovirulent human enterovirus 71 strain BJ110, which has a C4 genotype. Porcine small intestinal epithelial cells, peripheral blood mononuclear cells and neural cells were infected in vitro. Oral and combined oralnasal infection of 5-day-old neonatal gnotobiotic pigs with 5108 fluorescence forming units (FFU) resulted in shedding up to 18 days post-infection, with viral titers in rectal swab samples peaking at 2.22108 viral RNA copies/mL. Viral capsid proteins were detected in enterocytes within the small intestines on post-infection days (PIDs) 7 and 14. Additionally, viral RNA was detected in intestinal and extra-intestinal tissues, including the central nervous system, the lung and cardiac muscle. The infected neonatal gnotobiotic pigs developed fever, forelimb weakness, rapid breathing and some hand, foot and mouth disease symptoms. Flow cytometry analysis revealed increased frequencies of both CD4+ and CD8+ IFN-?-producing T cells in the brain and the blood on PID 14, but reduced frequencies were observed in the lung. Furthermore, high titers of serum virus-neutralizing antibodies were generated in both orally and combined oralnasally infected pigs on PIDs 7, 14, 21 and 28. Together, these results demonstrate that neonatal gnotobiotic pigs represent a novel animal model for evaluating vaccines for human enterovirus 71 and for understanding the pathogenesis of this virus and the associated immune responses.

Yang, Xingdong; Li, Guohua; Wen, Ke; Bui, Tammy; Liu, Fangning; Kocher, Jacob; Jortner, Bernard S; Vonck, Marlice; Pelzer, Kevin; Deng, Jie; Zhu, Runan; Li, Yuyun; Qian, Yuan; Yuan, Lijuan

2014-01-01

27

Recombination strategies and evolutionary dynamics of the Human enterovirus A global gene pool.  

PubMed

We analysed natural recombination in 79 Human enterovirus A strains representing 13 serotypes by sequencing of VP1, 2C and 3D genome regions. The half-life of a non-recombinant tree node in coxsackieviruses 2, 4 and 10 was only 3.5 years, and never more than 9 years. All coxsackieviruses that differed by more than 7?% of the nucleotide sequence in any genome region were recombinants relative to each other. Enterovirus 71 (EV71), on the contrary, displayed remarkable genetic stability. Three major EV71 clades were stable for 19-29 years, with a half-life of non-recombinant viruses between 13 and 18.5 years in different clades. Only five EV71 strains out of over 150 recently acquired non-structural genome regions from coxsackieviruses, while none of 80 contemporary coxsackieviruses had non-structural genes transferred from the three EV71 clades. In contrast to earlier observations, recombination between VP1 and 2C genome regions was not more frequent than between 2C and 3D regions. PMID:24425417

Lukashev, Alexander N; Shumilina, Elena Yu; Belalov, Ilya S; Ivanova, Olga E; Eremeeva, Tatiana P; Reznik, Vadim I; Trotsenko, O E; Drexler, Jan Felix; Drosten, Christian

2014-04-01

28

Antigenic and Genetic Diversity of Human Enterovirus 71 from 2009 to 2012, Taiwan  

PubMed Central

Different subgenogroups of enterovirus 71 (EV-71) have caused numerous outbreaks of hand, foot, and mouth disease worldwide, especially in the Asia-Pacific region. During the development of a vaccine against EV-71, the genetic and antigenic diversities of EV-71 isolates from Taiwan were analyzed by phylogenetic analyses and neutralization tests. The results showed that the dominant genogroups had changed twice, from B to C and from C to B, between 2009 and 2012. The subgenogroup B5 (B5b cluster) was dominant in 2008-2009 but was replaced by subgenogroup C4 in 2010-2011. From the end of 2011 to 2012, the re-emerging subgenogroup B5 (B5c cluster) was identified as the dominant subgenogroup of EV-71 outbreaks, and subgenogroups C2 and C4 were detected in sporadic cases. Interestingly, the amino acid substitution at position 145 in the VP1 gene was observed in some strains isolated from patients with acute flaccid paralysis. Furthermore, thirty-five strains and their corresponding serum samples were used to analyze the cross-protections and antigenic diversities among different subgenogroups (C4a, C5, B4, B5b, B5c, and C2-like) of EV-71. Evident antigenic diversity existed only for the C2-like subgenogroup, which was not effectively neutralized by other serum samples. In contrast, the anti-C2-like serum sample showed broad cross-reactivity against all other subgenogroups. Therefore, these results may provide valuable information for the selection of EV-71 vaccine candidates and the evolution of EV-71 subgenogroups in Taiwan from 2009 to 2012. PMID:24348916

Lin, Ting-Han; Wu, Ho-Sheng

2013-01-01

29

Seroepidemiology of human enterovirus71 and coxsackievirusA16 among children in Guangdong province, China  

PubMed Central

Background Hand, foot and mouth disease (HFMD) is a common pediatric illness. Mainly induced by the Enterovirus 71 and Coxsackievirus A 16 infections, the frequently occurred HFMD outbreaks have become a serious public health problem in Southeast Asia. Currently,only a few studies have investigated the human immunity to HFMD in China. In this study, we conducted a cohort study in Guangdong province, China. Methods Stored serum samples from children less than 10 years old were analyzed. The levels of EV71 and CA16 specific antibodies before, during and shortly after the 2008 large outbreak of HFMD were evaluated by the microneutralization test. The geometric mean titer (GMT) was calculated and compared. Statistical significance was taken as P?

2013-01-01

30

Development of A Sensitive and Specific Epitope-Blocking ELISA for Universal Detection of Antibodies to Human Enterovirus 71 Strains  

PubMed Central

Background Human Enterovirus 71 (EV71) is a common cause of hand, foot and mouth disease (HFMD) in young children. It is often associated with severe neurological diseases and mortalities in recent outbreaks across the Asia Pacific region. Currently, there is no efficient universal antibody test available to detect EV71 infections. Methodology/Principal Finding In the present study, an epitope-blocking ELISA was developed to detect specific antibodies to human EV71 viruses in human or animal sera. The assay relies on a novel monoclonal antibody (Mab 1C6) that specifically binds to capsid proteins in whole EV71 viruses without any cross reaction to any EV71 capsid protein expressed alone. The sensitivity and specificity of the epitope-blocking ELISA for EV71 was evaluated and compared to microneutralization using immunized animal sera to multiple virus genotypes of EV71 and coxsackieviruses. Further, 200 serum sample from human individuals who were potentially infected with EV71 viruses were tested in both the blocking ELISA and microneutralization. Results indicated that antibodies to EV71 were readily detected in immunized animals or human sera by the epitope blocking ELISA whereas specimens with antibodies to other enteroviruses yielded negative results. This assay is not only simpler to perform but also shows higher sensitivity and specificity as compared to microneutralization. Conclusion The epitope-blocking ELISA based on a unique Mab 1C6 provided highly sensitive and 100% specific detection of antibodies to human EV71 viruses in human sera. PMID:23383215

He, Fang; Kiener, Tanja K.; Lim, Xiao Fang; Tan, Yunrui; Raj, Kattur Venkatachalam Ashok; Tang, Manli; Chow, Vincent T. K.; Chen, Qingfeng; Kwang, Jimmy

2013-01-01

31

Recombinant tandem multi-linear neutralizing epitopes of human enterovirus 71 elicited protective immunity in mice  

PubMed Central

Background Human Enterovirus 71 (EV71) has emerged as the leading cause of viral encephalitis in children, especially in the Asia-Pacific regions. EV71 vaccine development is of high priority at present, and neutralization antibodies have been documented to play critical roles during in vitro and in vivo protection against EV71 infection. Results In this study, a novel strategy to produce EV71 vaccine candidate based on recombinant multiple tandem linear neutralizing epitopes (mTLNE) was proposed. The three well identified EV71 linear neutralizing epitopes in capsid proteins, VP1-SP55, VP1-SP70 and VP2-SP28, were sequentially linked by a Gly-Ser linker ((G4S)3), and expressed in E.coli in fusion with the Trx and His tag at either terminal. The recombinant protein mTLNE was soluble and could be purified by standard affinity chromatography. Following three dosage of immunization in adult mice, EV71-specific IgG and neutralization antibodies were readily induced by recombinant mTLNE. IgG subtyping demonstrated that lgG1 antibodies dominated the mTLNE-induced humoral immune response. Especially, cytokine profiling in spleen cells from the mTLNE-immunized mice revealed high production of IL-4 and IL-6. Finally, in vivo challenge experiments showed that passive transfer with anti-mTLNE sera conferred full protection against lethal EV71 challenge in neonatal mice. Conclusion Our results demonstrated that this rational designed recombinant mTLNE might have the potential to be further developed as an EV71 vaccine in the future. PMID:24885030

2014-01-01

32

Genetic Diversity of Human Enterovirus 68 Strains Isolated in Kenya Using the Hypervariable 3?- End of VP1 Gene  

PubMed Central

Reports of increasing worldwide circulation of human enterovirus-68 (EV68) are well documented. Despite health concerns posed by resurgence of these viruses, little is known about EV68 strains circulating in Kenya. In this study, we characterized 13 EV68 strains isolated in Kenya between 2008 and 2011 based on the Hypervariable 3?- end of the VP1 gene. Viral RNA was extracted from the isolates and partial VP1 gene amplified by RT-PCR, followed by nucleotide sequencing. Alignment of deduced amino acid sequences revealed substitutions in Kenyan EV68 isolates absent in the prototype reference strain (Fermon). The majority of these changes were present in the BC and DE-loop regions, which are associated with viral antigenicity and virulence. The Kenyan strains exhibited high sequence homology with respect to those from other countries. Natural selection analysis based on the VP1 region showed that the Kenyan EV68 isolates were under purifying selection. Phylogenetic analysis revealed that majority (84.6%) of the Kenyan strains belonged to clade A, while a minority belonged to clades B and C. Overall, our results illustrate that although EV68 strains isolated in Kenya were genetically and antigenically divergent from the prototype strain (Fermon), they were closely related to those circulating in other countries, suggesting worldwide transmissibility. Further, the presence of shared mutations by Kenyan EV68 strains and those isolated in other countries, indicates evolution in the VP1 region may be contributing to increased worldwide detection of the viruses. This is the first study to document circulation of EV68 in Kenya. PMID:25054861

Opanda, Silvanos M.; Wamunyokoli, Fred; Khamadi, Samoel; Coldren, Rodney; Bulimo, Wallace D.

2014-01-01

33

Enterovirus 2Apro Targets MDA5 and MAVS in Infected Cells  

PubMed Central

ABSTRACT RIG-I-like receptors (RLRs) MDA5 and RIG-I are key players in the innate antiviral response. Upon recognition of viral RNA, they interact with MAVS, eventually inducing type I interferon production. The interferon induction pathway is commonly targeted by viruses. How enteroviruses suppress interferon production is incompletely understood. MDA5 has been suggested to undergo caspase- and proteasome-mediated degradation during poliovirus infection. Additionally, MAVS is reported to be cleaved during infection with coxsackievirus B3 (CVB3) by the CVB3 proteinase 3Cpro, whereas MAVS cleavage by enterovirus 71 has been attributed to 2Apro. As yet, a detailed examination of the RLR pathway as a whole during any enterovirus infection is lacking. We performed a comprehensive analysis of crucial factors of the RLR pathway, including MDA5, RIG-I, LGP2, MAVS, TBK1, and IRF3, during infection of CVB3, a human enterovirus B (HEV-B) species member. We show that CVB3 inhibits the RLR pathway upstream of TBK1 activation, as demonstrated by limited phosphorylation of TBK1 and a lack of IRF3 phosphorylation. Furthermore, we show that MDA5, MAVS, and RIG-I all undergo proteolytic degradation in CVB3-infected cells through a caspase- and proteasome-independent manner. We convincingly show that MDA5 and MAVS cleavages are both mediated by CVB3 2Apro, while RIG-I is cleaved by 3Cpro. Moreover, we show that proteinases 2Apro and 3Cpro of poliovirus (HEV-C) and enterovirus 71 (HEV-A) exert the same functions. This study identifies a critical role of 2Apro by cleaving MDA5 and MAVS and shows that enteroviruses use a common strategy to counteract the interferon response in infected cells. IMPORTANCE Human enteroviruses (HEVs) are important pathogens that cause a variety of diseases in humans, including poliomyelitis, hand, foot, and mouth disease, viral meningitis, cardiomyopathy, and more. Like many other viruses, enteroviruses target the host immune pathways to gain replication advantage. The MDA5/MAVS pathway is responsible for recognizing enterovirus infections in the host cell and leads to expression of type I interferons (IFN-I), crucial antiviral signaling molecules. Here we show that three species of HEVs all employ the viral proteinase 2A (2Apro) to proteolytically target MDA5 and MAVS, leading to an efficient blockade upstream of IFN-I transcription. These observations suggest that MDA5/MAVS antagonization is an evolutionarily conserved and beneficial mechanism of enteroviruses. Understanding the molecular mechanisms of enterovirus immune evasion strategies will help to develop countermeasures to control infections with these viruses in the future. PMID:24390337

Feng, Qian; Langereis, Martijn A.; Lork, Marie; Nguyen, Mai; Hato, Stanleyson V.; Lanke, Kjerstin; Emdad, Luni; Bhoopathi, Praveen; Fisher, Paul B.; Lloyd, Richard E.

2014-01-01

34

Detection of Infectious Enteroviruses, Enterovirus Genomes, Somatic Coliphages, and Bacteroides fragilis Phages in Treated Wastewater  

PubMed Central

In this study, three types of treated wastewater were tested for infectious enteroviruses, the enterovirus genome, somatic coliphages, and Bacteroides fragilis phages. The aim of this work was to determine whether the presence of the two types of bacteriophages or of the enterovirus genome was a good indicator of infectious enterovirus contamination. The enterovirus genome was detected by reverse transcription-polymerase chain reaction. Infectious enteroviruses were quantified by cell culturing (BGM cells), and the bacteriophages were quantified by plaque formation on the host bacterium (Escherichia coli or B. fragilis) in agar medium. Forty-eight samples of treated wastewater were analyzed. Sixteen samples had been subjected to a secondary treatment for 8 to 12 h (A), 16 had been subjected to a secondary treatment for 30 h (B1), and 16 had been subjected to both secondary and tertiary treatments (B2). The mean concentrations of somatic coliphages were 4.9 104 PFU liter?1 for treatment line A, 9.8 103 PFU liter?1 for B1, and 1.4 103 PFU liter?1 for B2, with all the samples testing positive (100%). The mean concentrations of B. fragilis phages were 1.7 103 PFU liter?1 for A (100% positive samples), 17 to 24 PFU liter?1 for B1 (44% positive samples), and 0.8 to 13 PFU liter?1 for B2 (6% positive samples). The mean concentrations of infectious enteroviruses were 4 most probable number of cytopathogenic units (MPNCU) liter?1 for A (31% positive samples) and <1 MPNCU liter?1 for B1 and B2 (0% positive samples). The percentages of samples testing positive for the enterovirus genome were 100% for A, 56% for B1, and 19% for B2. The percentages of samples testing positive for the enterovirus genome were significantly higher than those for infectious enteroviruses. This finding may have been due to the presence of noninfectious enteroviruses or to the presence of infectious enteroviruses that do not multiply in BGM cell cultures. However, under our experimental conditions, nondetection of the genome implies the absence of infectious viruses. There was a significant correlation between the concentration of somatic coliphages or B. fragilis phages and the presence of infectious enteroviruses or the presence of the enterovirus genome. However, the somatic coliphage concentration did not lead to fluctuations in the infectious enterovirus concentration, whereas the B. fragilis phage concentration did. PMID:9797281

Gantzer, C.; Maul, A.; Audic, J. M.; Schwartzbrod, L.

1998-01-01

35

Elucidating the host-pathogen interaction between human colorectal cells and invading Enterovirus 71 using transcriptomics profiling  

PubMed Central

Enterovirus 71 (EV71) is one of the main etiological agents for Hand, Foot and Mouth Disease (HFMD) and has been shown to be associated with severe clinical manifestation. Currently, there is no antiviral therapeutic for the treatment of HFMD patients owing to a lack of understanding of EV71 pathogenesis. This study seeks to elucidate the transcriptomic changes that result from EV71 infection. Human whole genome microarray was employed to monitor changes in genomic profiles between infected and uninfected cells. The results reveal altered expression of human genes involved in critical pathways including the immune response and the stress response. Together, data from this study provide valuable insights into the hostpathogen interaction between human colorectal cells and EV71. PMID:24918057

Lui, Yan Long Edmund; Tan, Tuan Lin; Timms, Peter; Hafner, Louise Marie; Tan, Kian Hwa; Tan, Eng Lee

2014-01-01

36

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

2013-01-01

37

Evaluation of methods using celite to concentrate norovirus, adenovirus and enterovirus from wastewater.  

PubMed

Enteroviruses, noroviruses and adenoviruses are among the most common viruses infecting humans worldwide. These viruses are shed in the feces of infected individuals and can accumulate in wastewater, making wastewater a source of a potentially diverse group of enteric viruses. In this study, two procedures were evaluated to concentrate noroviruses, adenoviruses and enteroviruses from primary effluent of wastewater. In the first procedure, indigenous enteroviruses, noroviruses and adenoviruses were concentrated using celite (diatomaceous earth) followed by centrifugation through a 30K MWCO filter and nucleic acid extraction. The second procedure used celite concentration followed by nucleic acid extraction only. Virus quantities were measured using qPCR. A second set of primary effluent samples were seeded with Coxsackievirus A7, Coxsackievirus B1, poliovirus 1 or enterovirus 70 before concentration and processed through both procedures for recovery evaluation of enterovirus species representatives. The pairing of the single step extraction procedure with the celite concentration process resulted in 47-98% recovery of examined viruses, while the celite concentration process plus additional centrifugal concentration before nucleic acid extraction showed reduced recovery (14-47%). The celite concentration process followed by a large volume nucleic acid extraction technique proved to be an effective procedure for recovering these important human pathogens from wastewater. PMID:23727118

Brinkman, Nichole E; Haffler, Tyler D; Cashdollar, Jennifer L; Rhodes, Eric R

2013-10-01

38

Mechanisms and consequences of enterovirus persistence in cardiac myocytes and cells of the immune system  

Microsoft Academic Search

In humans and experimental murine models enteroviruses, and in particular coxsackieviruses of group B (CVB), may induce chronic myocarditis associated with a persistent type of heart muscle infection. Persistent myocardial infection has been characterized by restricted viral replication and gene expression, which is capable of sustaining chronic inflammation. Altered replication and transcription of the virus, in addition to an immune

Reinhard Kandolf; Martina Sauter; Christian Aepinus; Jens-Jrg Schnorr; Hans-Christoph Selinka; Karin Klingel

1999-01-01

39

Enterovirus-71 Virus-Like Particles Induce the Activation and Maturation of Human Monocyte-Derived Dendritic Cells through TLR4 Signaling  

PubMed Central

Enterovirus 71 (EV71) causes seasonal epidemics of hand-foot-and-mouth disease and has a high mortality rate among young children. We recently demonstrated potent induction of the humoral and cell-mediated immune response in monkeys immunized with EV71 virus-like particles (VLPs), with a morphology resembling that of infectious EV71 virions but not containing a viral genome, which could potentially be safe as a vaccine for EV71. To elucidate the mechanisms through which EV71 VLPs induce cell-mediated immunity, we studied the immunomodulatory effects of EV71 VLPs on human monocyte-derived dendritic cells (DCs), which bind to and incorporate EV71 VLPs. DC treatment with EV71 VLPs enhanced the expression of CD80, CD86, CD83, CD40, CD54, and HLA-DR on the cell surface; increased the production of interleukin (IL)-12 p40, IL-12 p70, and IL-10 by DCs; and suppressed the capacity of DCs for endocytosis. Treatment with EV71 VLPs also enhanced the ability of DCs to stimulate nave T cells and induced secretion of interferon (IFN)-? by T cells and Th1 cell responses. Neutralization with antibodies against Toll-like receptor (TLR) 4 suppressed the capacity of EV71 VLPs to induce the production of IL-12 p40, IL-12 p70, and IL-10 by DCs and inhibited EV71 VLPs binding to DCs. Our study findings clarified the important role for TLR4 signaling in DCs in response to EV71 VLPs and showed that EV71 VLPs induced inhibitor of kappaB alpha (I?B?) degradation and nuclear factor of kappaB (NF-?B) activation. PMID:25360749

Lin, Yu-Li; Hu, Yu-Chen; Liang, Cheng-Chao; Lin, Shih-Yeh; Liang, Yu-Chih; Yuan, Hui-Ping; Chiang, Bor-Luen

2014-01-01

40

Occurrence of enteroviruses in community swimming pools.  

PubMed Central

Municipal swimming pools and wading pools were examined for the presence of human enteric viruses using a portable virus concentrator at the site to concentrate viruses from 100-gallon to 500-gallon samples. Ten of 14 samples contained viruses; three of these were positive for virus in the presence of residual free chlorine. Enteroviruses were isolated from two pools which exceeded the 0.4 ppm free residual chlorine standard. This study appears to be supportive of recent evidence that indicates a higher incidence of enterovirus infection among bathers. All seven wading pool samples contained virus. Coxsackieviruses B3 and B4, poliovirus 1, and echovirus 7 were isolated. Total coliform bacteria were not adequate indicators of the presence of virus, as six of the samples were positive for virus but negative for coliforms. Total plate counts appeared to provide a better indication of the sanitary quality of the pool water, but viruses could still be detected in samples that met currently recommended bacterial levels. It is possible that swimming and wading pools may serve as a means of transmission of enteroviral disease, especially in children, during summer months. PMID:6267950

Keswick, B H; Gerba, C P; Goyal, S M

1981-01-01

41

Enterovirus 68 Overview  

MedlinePLUS

... be accessed by typing cdc.gov/non-polio-enterovirus in most web browsers. While the website provides general information about non-polio enteroviruses, the section States with lab-confirmed EV-D68 infection provides a state-by-state tracking of ...

42

[Acute enterovirus uveitis in infants].  

PubMed

Enterovirus uveitis (EU) is a new infant eye disease that was first detected and identified in Russia in 1980-1981. Three subtypes of human echoviruses (EV19K, EV11A, and EV11/B) caused 5 nosocomial outbreaks of EU in different Siberian cities and towns in 1980-1989, by affecting more than 750 children mainly below one year of age. Sporadic and focal EU cases (more than 200) were also retrospectively diagnosed in other regions of Russia and in different countries of the former Soviet Union. There were following clinical manifestations: common symptoms of the infection; acute uveitis (rapid focal iridic destruction, pupillary deformities, formation of membranes in the anterior chamber of the eye); and in 15-30% of cases severe complications, cataract, glaucoma, vision impairments. Uveitis strains EV19 and EV11 caused significant uveitis in primates after inoculation into the anterior chamber of the eye, as well as sepsis-like fatal disease with liver necrosis after venous infection. The uveitis strains are phylogenetically and pathogenetically close for primates to strains EV19 and EV11 isolated from young children with sepsis-like disease. The contents of this review have been published in the Reviews in Medical Virology, 2004, vol. 14, p. 241-254. PMID:16078433

Lashkevich, V A; Koroleva, G A; Lukashev, A N; Denisova, E V; Katargina, L A; Khoroshilova-Maslova, I P

2005-01-01

43

Human enteroviruses are not the cause of neurological impairments in children at the Korle-Bu Teaching Hospital  

PubMed Central

Introduction Convulsions associated with fever and acute onset of unknown aetiology with case fatalities have become a long observed medical condition at the Child Health Department of the Korle-Bu Teaching Hospital. Children admitted to the department with seizures of undetermined origin and fever has been a source of diagnostic confusion. Studies from the Asia Pacific region suggest a link with non-polio enteroviruses. The aim of the study was to investigate the association between non-polio enterovirus and acute encephalopathy causing neurological morbidity in children. Methods One hundred and fifty cerebrospinal fluid (CSF), throat swab and serum samples were collected from participants at the Child Health Department of the Korle-Bu Teaching Hospital for virus isolation and characterization. Samples were cultured on cells and positive culture assayed by microneutralisation. Direct PCR as well as multiplex PCR were used to detect other viral agents present. Results Enterovirus isolation rate was approximately 0.67%. Intratypic differentiation by molecular characterization identified a poliovirus from vaccine origin. Further screening by real-time RT-PCR identified the virus as normal Sabin and not vaccine-derive poliovirus. No arbovirus was however detected. Conclusion Non-polio enteroviruses and chikugunya virus were found not to be the etiologic agent responsible for the convulsion with neurologic morbidity observed in the Ghanaian children. Investigation for other viral agents is recommended.

Tettey, Prudence; Badoe, Ebenezer; Adiku, Theophilus; Obodai, Eva; Odoom, John Kofi

2014-01-01

44

Infectious Diseases and Tropical Disease Pathology: SS16-1 UNDERSTANDING ENTEROVIRUS 71 INFECTION AND NEUROPATHOGENESIS: PERSPECTIVE FROM HUMAN AND ANIMAL MODEL STUDIES.  

PubMed

Enterovirus 71 (EV71) (family Picornaviridae, genus Human Enterovirus A) causes the relatively mild hand, foot and mouth disease in children. However, rare complications including meningitis, acute transverse myelitis, flaccid paralysis and encephalomyelitis may occur. Recently, many EV71 outbreaks with significant mortality in the Asia-Pacific region and beyond have posed serious public health concerns worldwide. Fatal encephalomyelitis is mainly the result of severe inflammation and neuronal damage in the brainstem, spinal cord and other parts of the brain. Medullary involvement is believed to result in neurogenic pulmonary oedema and sudden collapse typically found in fatal cases. A recent study has demonstrated viral replication in tonsillar crypt epithelium, suggesting that tonsil may be a portal for viral entry into the body. Presumably this is followed by viral replication in other sites (e.g. skin) and/or viraemia, leading to neuroinvasion possibly by a retrograde peripheral motor nerve route. There is also increasing evidence that the SCARB2 receptor may be an important receptor for human EV71 infection. A possible hypothesis for viral transmission into the body leading to CNS infection will be discussed. The value of human material and animal models to study viral infections in general and EV71 infection in particular is emphasized. PMID:25188114

Wong, Kum Thong; ONg, Kien Chai

2014-10-01

45

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

2003-01-01

46

An emerging recombinant human enterovirus 71 responsible for the 2008 outbreak of hand foot and mouth disease in Fuyang city of China.  

PubMed

Hand, foot and mouth disease (HFMD), a common contagious disease that usually affects children, is normally mild but can have life-threatening manifestations. It can be caused by enteroviruses, particularly Coxsackieviruses and human enterovirus 71 (HEV71) with highly variable clinical manifestations. In the spring of 2008, a large, unprecedented HFMD outbreak in Fuyang city of Anhui province in the central part of southeastern China resulted in a high aggregation of fatal cases. In this study, epidemiologic and clinical investigations, laboratory testing, and genetic analyses were performed to identify the causal pathogen of the outbreak. Of the 6,049 cases reported between 1 March and 9 May of 2008, 3023 (50%) were hospitalized, 353 (5.8%) were severe and 22 (0.36%) were fatal. HEV71 was confirmed as the etiological pathogen of the outbreak. Phylogenetic analyses of entire VP1 capsid protein sequence of 45 Fuyang HEV71 isolates showed that they belong to C4a cluster of the C4 subgenotype. In addition, genetic recombinations were found in the 3D region (RNA-dependent RNA polymerase, a major component of the viral replication complex of the genome) between the Fuyang HEV71 strain and Coxsackievirus A16 (CV-A16), resulting in a recombination virus. In conclusion, an emerging recombinant HEV71 was responsible for the HFMD outbreak in Fuyang City of China, 2008. PMID:20459851

Zhang, Yan; Zhu, Zhen; Yang, Weizhong; Ren, Jun; Tan, Xiaojuan; Wang, Yu; Mao, Naiying; Xu, Songtao; Zhu, Shuangli; Cui, Aili; Zhang, Yong; Yan, Dongmei; Li, Qun; Dong, Xiaoping; Zhang, Jing; Zhao, Yueping; Wan, Junfeng; Feng, Zijian; Sun, Junling; Wang, Shiwen; Li, Dexin; Xu, Wenbo

2010-01-01

47

Human adenovirus (HAdV), human enterovirus (hEV), and genogroup A rotavirus (GARV) in tap water in southern Brazil.  

PubMed

The effects of viral gastroenteritis are more devastating in children than in any other age category. Thus, children exposed to the consumption of low quality water are at an increased risk of infection, especially in regions where sanitation is inadequate. The present study aimed to provide a survey of the occurrence of representative enteric viruses: human adenovirus (HAdV), human enteroviruses (hEV), and genogroup A rotavirus (GARV) in tap water samples collected in public schools located at six municipalities of Rio Grande do Sul, southern Brazil. Seventy-three schools were included in the study and tap water samples were analyzed by conventional PCR for the presence of HAdV, hEV, and GARV genomes. hEV showed the highest detection rate (27.4%), followed by HAdV (23.3%), and GARV (16.4%). New approaches to water monitoring should be considered to promote a better water quality and reduce the risk of waterborne diseases, especially considering drinking water to be served to vulnerable individuals. PMID:25252356

Kluge, M; Fleck, J D; Soliman, M C; Luz, R B; Fabres, R B; Comerlato, J; Silva, J V S; Staggemeier, R; Vecchia, A D; Capalonga, R; Oliveira, A B; Henzel, A; Rigotto, C; Spilki, F R

2014-09-01

48

Antigenic Diversity of Enteroviruses Associated with Nonpolio Acute Flaccid Paralysis, India, 2007-2009  

PubMed Central

Because of the broadened acute flacid paralysis (AFP) definition and enhanced surveillance, many nonpolio AFP (NP-AFP) cases have been reported in India since 2005. To determine the spectrum of nonpolio enterovirus (NPEV) serotypes associated with NP-AFP from polio-endemic and -free regions, we studied antigenic diversity of AFP-associated NPEVs. Of fecal specimens from 2,786 children with NP-AFP in 1 polio-endemic and 2 polio-free states, 823 (29.5%) were positive for NPEVs in RD cells, of which 532 (64.6%) were positive by viral protein 1 reverse transcription PCR. We identified 66 serotypes among 581 isolates, with enterovirus 71 most frequently (8.43%) detected, followed by enterovirus 13 (7.1%) and coxsackievirus B5 (5.0%). Most strains within a serotype represented new genogropups or subgenogroups. Agents for ?35.0% and 70.0% of culture-positive and -negative cases, respectively, need to be identified. Association of human enterovirus with NP-AFP requires better assessment and understanding of health risks of NPEV infections after polio elimination. PMID:23092622

Yergolkar, Prasanna; Shankarappa, K. Subbanna

2012-01-01

49

Molecular Typing of Enteroviruses: Current Status and Future Requirements  

PubMed Central

Human enteroviruses have traditionally been typed according to neutralization serotype. This procedure is limited by the difficulty in culturing some enteroviruses, the availability of antisera for serotyping, and the cost and technical complexity of serotyping procedures. Furthermore, the impact of information derived from enterovirus serotyping is generally perceived to be low. Enteroviruses are now increasingly being detected by PCR rather than by culture. Classical typing methods will therefore no longer be possible in most instances. An alternative means of enterovirus typing, employing PCR in conjunction with molecular genetic techniques such as nucleotide sequencing or nucleic acid hybridization, would complement molecular diagnosis, may overcome some of the problems associated with serotyping, and would provide additional information regarding the epidemiology and biological properties of enteroviruses. We argue the case for developing a molecular typing system, discuss the genetic basis of such a system, review the literature describing attempts to identify or classify enteroviruses by molecular methods, and suggest ways in which the goal of molecular typing may be realized. PMID:9457433

Muir, Peter; Kmmerer, Ulrike; Korn, Klaus; Mulders, Mick N.; Pyry, Tuija; Weissbrich, Benedikt; Kandolf, Reinhard; Cleator, Graham M.; van Loon, Anton M.

1998-01-01

50

A Sabin 2-related poliovirus recombinant contains a homologous sequence of human enterovirus species C in the viral polymerase coding region.  

PubMed

A type 2 vaccine-related poliovirus (strain CHN3024), differing from the Sabin 2 strain by 0.44% in the VP1 coding region was isolated from a patient with vaccine-associated paralytic poliomyelitis. Sequences downstream of nucleotide position 6735 (3D(pol) coding region) were derived from an unidentified sequence; no close match for a potential parent was found, but it could be classified into a non-polio human enteroviruses species C (HEV-C) phylogeny. The virus differed antigenically from the parental Sabin strain, having an amino acid substitution in the neutralizing antigenic site 1. The similarity between CHN3024 and Sabin 2 sequences suggests that the recombination was recent; this is supported by the estimation that the initiating OPV dose was given only 36-75 days before sampling. The patient's clinical manifestations, intratypic differentiation examination, and whole-genome sequencing showed that this recombinant exhibited characteristics of neurovirulent vaccine-derived polioviruses (VDPV), which may, thus, pose a potential threat to a polio-free world. PMID:19946714

Zhang, Yong; Zhang, Fan; Zhu, Shuangli; Chen, Li; Yan, Dongmei; Wang, Dongyan; Tang, Ruiyan; Zhu, Hui; Hou, Xiaohui; An, Hongqiu; Zhang, Hong; Xu, Wenbo

2010-02-01

51

Complete genome sequence analysis of an enterovirus 75 isolate from China.  

PubMed

Enterovirus 75 (EV-B75) is a member of the species Enterovirus B (EV-B). So far, only the complete genome of the prototype strain from the United States is available. Here, we report the genome sequence of an EV-B75 isolate from an acute flaccid paralysis patient in China. Sequence analysis revealed high nucleotide sequence divergence from foreign EV-B75 strains and suggested several recombination events with other serotypes of EV-B. PMID:24852072

Tao, Zexin; Cui, Ning; Lin, Xiaojuan; Wang, Haiyan; Song, Lizhi; Li, Yan; Zhang, Li; Liu, Xiaolin; Wang, Suting; Xu, Aiqiang; Song, Yanyan

2014-10-01

52

Assessment of the risks for human health of adenoviruses, hepatitis a virus, rotaviruses and enteroviruses in the buffalo river and three source water dams in the eastern cape.  

PubMed

Buffalo River is an important water resource in the Eastern Cape Province of South Africa. The potential risks of infection constituted by exposure to human enteric viruses in the Buffalo River and three source water dams along its course were assessed using mean values and static quantitative microbial risk assessment (QMRA). The daily risks of infection determined by the exponential model [for human adenovirus (HAdV) and enterovirus (EnV)] and the beta-Poisson model (for hepatitis A virus (HAV) and rotavirus (RoV)) varied with sites and exposure scenario. The estimated daily risks of infection values at the sites where the respective viruses were detected, ranged from 7.31נ10(-3) to 1 (for HAdV), 4.23נ10(-2) to 6.54נ10(-1) (RoV), 2.32נ10(-4) to 1.73נ10(-1) (HAV) and 1.32נ10(-4) to 5.70נ10(-2) (EnV). The yearly risks of infection in individuals exposed to the river/dam water via drinking, recreational, domestic or irrigational activities were unacceptably high, exceeding the acceptable risk of 0.01% (10(-4) infection/person/year), and the guideline value used as by several nations for drinking water. The risks of illness and death from infection ranged from 6.58נ10(-5) to 5.0נ10(-1) and 6.58נ10(-9) to 5.0נ10(-5), respectively. The threats here are heightened by the high mortality rates for HAV, and its endemicity in South Africa. Therefore, we conclude that the Buffalo River and its source water dams are a public health hazard. The QMRA presented here is the first of its kinds in the Eastern Cape Province and provides the building block for a quantitatively oriented local guideline for water quality management in the Province. PMID:24676673

Chigor, Vincent N; Sibanda, Timothy; Okoh, Anthony I

2014-06-01

53

Enterovirus type 70-neutralizing IgM in animal sera.  

PubMed

The neutralizing activity against human enterovirus type 70 was found in serum samples from normal cattle, horses, sheep, goats, swine and chickens raised in Japan. The frequency was variable depending upon animal species and the year of bleeding. The neutralizing activity in bovine sera was shown to reside in IgM by sucrose gradient centrifugation and immune gel electrophoresis. These findings suggested that the neutralizing substance in domestic animal sera is the antibody of IgM class elicited by unidentified viruses antigenically related to human enterovirus type 70. PMID:6287063

Sasagawa, A; Miyamura, K; Kono, R

1982-04-01

54

Binding of Glutathione to Enterovirus Capsids Is Essential for Virion Morphogenesis  

PubMed Central

Enteroviruses (family of the Picornaviridae) cover a large group of medically important human pathogens for which no antiviral treatment is approved. Although these viruses have been extensively studied, some aspects of the viral life cycle, in particular morphogenesis, are yet poorly understood. We report the discovery of TP219 as a novel inhibitor of the replication of several enteroviruses, including coxsackievirus and poliovirus. We show that TP219 binds directly glutathione (GSH), thereby rapidly depleting intracellular GSH levels and that this interferes with virus morphogenesis without affecting viral RNA replication. The inhibitory effect on assembly was shown not to depend on an altered reducing environment. Using TP219, we show that GSH is an essential stabilizing cofactor during the transition of protomeric particles into pentameric particles. Sequential passaging of coxsackievirus B3 in the presence of low GSH-levels selected for GSH-independent mutants that harbored a surface-exposed methionine in VP1 at the interface between two protomers. In line with this observation, enteroviruses that already contained this surface-exposed methionine, such as EV71, did not rely on GSH for virus morphogenesis. Biochemical and microscopical analysis provided strong evidence for a direct interaction between GSH and wildtype VP1 and a role for this interaction in localizing assembly intermediates to replication sites. Consistently, the interaction between GSH and mutant VP1 was abolished resulting in a relocalization of the assembly intermediates to replication sites independent from GSH. This study thus reveals GSH as a novel stabilizing host factor essential for the production of infectious enterovirus progeny and provides new insights into the poorly understood process of morphogenesis. PMID:24722756

Thibaut, Hendrik Jan; Thys, Bert; Canela, Mara-Dolores; Aguado, Leire; Wimmer, Eckard; Paul, Aniko; Prez-Prez, Mara-Jess; van Kuppeveld, Frank J. M.; Neyts, Johan

2014-01-01

55

Identification of 20 common human enterovirus serotypes by use of a reverse transcription-PCR-based reverse line blot hybridization assay.  

PubMed

The more than 100 human enterovirus (HEV) serotypes can also be classified into four species, HEV-A to -D, based on phylogenetic analysis of multiple gene regions. Current molecular typing methods depend largely on reverse transcription-PCR (RT-PCR) amplification and nucleotide sequencing of the entire or 3' half of the VP1 gene. An RT-PCR-based reverse line blot (RLB) hybridization assay was developed as a rapid and efficient approach to characterize common HEVs. Twenty HEV serotypes accounted for 87.1% of all HEVs isolated at an Australian reference virology laboratory from 1979 to 2007. VP1 sequences of all known HEV prototype strains were aligned to design one sense primer and three antisense primers for RT-PCR. After sequencing of the complete VP1 genes of 37 previously serotyped examples of the commonest 20 serotypes and alignment of these VP1 sequences with GenBank sequences, four serotype-specific probes for each serotype were designed for RLB. The RT-PCR-RLB assay was then applied to 132 HEV isolates, made up of the previously sequenced 37 isolates and another 95 serotyped clinical isolates. The RT-PCR-RLB genotypes corresponded with the serotypes for 131/132 isolates; the one exception was confirmed by VP1 sequencing, and the genotype was confirmed by repeat conventional serotyping. Genotyping by RT-PCR-RLB complements traditional serotyping methods and VP1 sequencing and has the advantages of convenience, speed, and accuracy. RT-PCR-RLB allows detection of specific enteroviral serotypes or genotypes associated with HEV outbreaks and significant disease. PMID:19571022

Zhou, Fei; Kong, Fanrong; McPhie, Kenneth; Ratnamohan, Mala; Donovan, Linda; Zeng, Frank; Gilbert, Gwendolyn L; Dwyer, Dominic E

2009-09-01

56

Evaluation of a direct reverse transcription loop-mediated isothermal amplification method without RNA extraction for the detection of human enterovirus 71 subgenotype C4 in nasopharyngeal swab specimens.  

PubMed

Human enterovirus 71 (EV71) is the major causative agent of hand, foot, and mouth disease (HFMD) worldwide and has been associated with neurological complications which resulted in fatalities during recent outbreak in Asia pacific region. A direct reverse transcription loop-mediated isothermal amplification (direct RT-LAMP) assay using heat-treated samples without RNA extraction was developed and evaluated for the detection of EV71 subgenotype C4 in nasopharyngeal swab specimens. The analytical sensitivity and specificity of the direct RT-LAMP assay were examined. The detection limit of the direct RT-LAMP assays was 1.6 of a 50% tissue culture infective dose (TCID(50)) per reaction and no cross-reaction was observed with control viruses including Cosackievirus A (CVA) viruses (CVA2,4,5,7,9,10,14,16, and 24), Coxsackievirus B (CVB) viruses (CVB1,2,3,4, and 5) or ECHO viruses (ECHO3,6,11, and 19). The direct RT-LAMP assay was evaluated and compared to both RT-LAMP and quantitative real-time PCR (qRT-PCR) in detecting EV71 infection with 145 nasopharyngeal swab specimens. The clinical performance demonstrated the sensitivity and specificity of direct RT-LAMP was reported to be 90.3% and 100% respectively, compared to RT-LAMP, and 86.83% and 100% respectively, compared to qRT-PCR. These data demonstrated that the direct RT-LAMP assay can potentially be developed for the point of care screening of EV71 infection in China. PMID:23272248

Nie, Kai; Qi, Shun-Xiang; Zhang, Yong; Luo, Le; Xie, Yun; Yang, Meng-Jie; Zhang, Yi; Li, Jin; Shen, Hongwei; Li, Qi; Ma, Xue-Jun

2012-01-01

57

Evaluation of a Direct Reverse Transcription Loop-Mediated Isothermal Amplification Method without RNA Extraction for the Detection of Human Enterovirus 71 Subgenotype C4 in Nasopharyngeal Swab Specimens  

PubMed Central

Human enterovirus 71 (EV71) is the major causative agent of hand, foot, and mouth disease (HFMD) worldwide and has been associated with neurological complications which resulted in fatalities during recent outbreak in Asia pacific region. A direct reverse transcription loop-mediated isothermal amplification (direct RT-LAMP) assay using heat-treated samples without RNA extraction was developed and evaluated for the detection of EV71 subgenotype C4 in nasopharyngeal swab specimens. The analytical sensitivity and specificity of the direct RT-LAMP assay were examined. The detection limit of the direct RT-LAMP assays was 1.6 of a 50% tissue culture infective dose (TCID50) per reaction and no cross-reaction was observed with control viruses including Cosackievirus A (CVA) viruses (CVA2,4,5,7,9,10,14,16, and 24), Coxsackievirus B (CVB) viruses (CVB1,2,3,4, and 5) or ECHO viruses (ECHO3,6,11, and 19). The direct RT-LAMP assay was evaluated and compared to both RT-LAMP and quantitative real-time PCR (qRT-PCR) in detecting EV71 infection with 145 nasopharyngeal swab specimens. The clinical performance demonstrated the sensitivity and specificity of direct RT-LAMP was reported to be 90.3% and 100% respectively, compared to RT-LAMP, and 86.83% and 100% respectively, compared to qRT-PCR. These data demonstrated that the direct RT-LAMP assay can potentially be developed for the point of care screening of EV71 infection in China. PMID:23272248

Luo, Le; Xie, Yun; Yang, Meng-jie; Zhang, Yi; Li, Jin; Shen, Hongwei; Li, Qi; Ma, Xue-jun

2012-01-01

58

Evidence for an enterovirus as the cause of encephalitis lethargica  

PubMed Central

Background The epidemic of encephalitis lethargica (EL), called classical EL, was rampant throughout the world during 19171926, affecting half a million persons. The acute phase was lethal for many victims. Post-encephalitic parkinsonism (PEP) affected patients for decades. Our purpose was to investigate the cause of classical EL by studying the few available brain specimens. Cases of PEP and modern EL were also studied. Transmission electron microscopy (TEM) and immunohistochemistry were employed to examine brain from four classical EL cases, two modern EL cases and one PEP case. Methods Standard methods for TEM, immunohistochemistry and RTPCR were applied. Results 27?nm virus-like particles (VLP) were observed in the cytoplasm and nuclei of midbrain neurons in all classical EL cases studied. Large (50?nm) VLP and 27?nm intranuclear VLP were observed in the modern EL cases and the PEP case. Influenza virus particles were not found. VLP were not observed in the control cases. TEM of cell cultures inoculated with coxsackievirus B4 and poliovirus revealed both small and large intranuclear virus particles and small cytoplasmic particles, similar to the VLP in EL neurons. In the EL brains, nascent VLP were embedded in putative virus factories and on endoplasmic reticulum (ER). The VLP in the cases of classical EL survived, whereas ribosomes underwent autolysis due to the lack of refrigeration and slow formaldehyde fixation of whole brain. The VLP were larger than ribosomes from well preserved brain. Immunohistochemistry of classical EL cases using anti-poliovirus and anti-coxsackievirus B polyclonal antibodies showed significant staining of cytoplasm and nuclei of neurons as well as microglia and neuropil. Purkinje cells were strongly stained. A 97-bp RNA fragment of a unique virus was isolated from brain tissue from acute EL case #91558. Sequence analysis revealed up to 95% identity to multiple human Enteroviruses. Additional cases had Enterovirus positive reactions by real time PCR. Conclusions The data presented here support the hypothesis that the VLP observed in EL tissue is an Enterovirus. PMID:22715890

2012-01-01

59

CLINICAL SEVERITY OF RHINOVIRUS/ENTEROVIRUS COMPARED TO OTHER RESPIRATORY VIRUSES IN CHILDREN.  

E-print Network

?? Background: Human rhinovirus/enterovirus (HRV/ENT) infections are commonly identified in children with acute respiratory infections (ARIs), but data on their clinical severity remains limited. We (more)

Asner, Sandra Andrea, M.D

2013-01-01

60

Improved detection limit in rapid detection of human enterovirus 71 and coxsackievirus A16 by a novel reverse transcription-isothermal multiple-self-matching-initiated amplification assay.  

PubMed

Rapid detection of human enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) is important in the early phase of hand-foot-and-mouth disease (HFMD). In this study, we developed and evaluated a novel reverse transcription-isothermal multiple-self-matching-initiated amplification (RT-IMSA) assay for the rapid detection of EV71 and CVA16 by use of reverse transcriptase, together with a strand displacement DNA polymerase. Real-time RT-IMSA assays using a turbidimeter and visual RT-IMSA assays to detect EV71 and CVA16 were established and completed in 1 h, and the reported corresponding real-time reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assays targeting the same regions of the VP1 gene were adopted as parallel tests. Through testing VP1 RNAs transcribed in vitro, the real-time RT-IMSA assays exhibited better linearity of quantification, with R(2) values of 0.952 (for EV71) and 0.967 (for CVA16), than the real-time RT-LAMP assays, which had R(2) values of 0.803 (for EV71) and 0.904 (for CVA16). Additionally, the detection limits of the real-time RT-IMSA assays (approximately 937 for EV71 and 67 for CVA16 copies/reaction) were higher than those of real-time RT-LAMP assays (approximately 3,266 for EV71 and 430 for CVA16 copies/reaction), and similar results were observed in the visual RT-IMSA assays. The new approaches also possess high specificities for the corresponding targets, with no cross-reactivity observed. In clinical assessment, compared to commercial reverse transcription-quantitative PCR (qRT-PCR) kits, the diagnostic sensitivities of the real-time RT-IMSA assays (96.4% for EV71 and 94.6% for CVA16) were higher than those of the real-time RT-LAMP assays (91.1% for EV71 and 90.8% for CVA16). The visual RT-IMSA assays also exhibited the same results. In conclusion, this proof-of-concept study suggests that the novel RT-IMSA assay is superior to the RT-LAMP assay in terms of detection limit and has the potential to rapidly detect EV71 and CVA16 viruses. PMID:24648558

Ding, Xiong; Nie, Kai; Shi, Lei; Zhang, Yong; Guan, Li; Zhang, Dan; Qi, Shunxiang; Ma, Xuejun

2014-06-01

61

Structure determination of enterovirus 71  

SciTech Connect

Enterovirus 71 is a picornavirus that causes hand, foot and mouth disease but may induce fatal neurological illness in infants and young children. Enterovirus 71 crystallized in a body-centered orthorhombic space group with two particles in general orientations in the crystallographic asymmetric unit. Determination of the particle orientations required that the locked rotation function excluded the twofold symmetry axes from the set of icosahedral symmetry operators. This avoided the occurrence of misleading high rotation-function values produced by the alignment of icosahedral and crystallographic twofold axes. Once the orientations and positions of the particles had been established, the structure was solved by molecular replacement and phase extension.

Plevka, Pavel; Perera, Rushika; Cardosa, Jane; Kuhn, Richard J.; Rossmann, Michael G. (Purdue); (Sentinext)

2013-02-20

62

[Social and economic significance of enterovirus infection and its role in etiologic structure of infectious diseases in the world].  

PubMed

Human enteroviruses comprised by more than 100 serotypes, they spread everywhere and can cause wide spectrum of diseases as well as significant social and economic loss. Influenza-like illness and mild forms of enterovirus infection (herpangina, exanthema) are widespread and causes of significant number of visits in clinics. Economic cost of mild form of enterovirus infection is not high although great number of cases (10 - 15 mln cases yearly in USA) determines its important economic significance. Single cases and outbreaks of enterovirus aseptic meningitis occur less frequently but lead to significant economic burden due to hospitalization costs. Enteroviruses are also cause up to 30% of sepsis-like disease in newborns and play important role in infant morbidity and mortality. Potential of enteroviruses as a source of new diseases in humans has a special significance for practical healthcare. In XX century enteroviruses became a cause of pandemics of paralytic poliomyelitis, hemorrhagic conjunctivitis, and foot-and-mouth-like disease, which caused vast social and economic loss, and emergence of new forms of enterovirus infection is quite possible in XXI century. PMID:21061587

Lukashev, A N; Ivanova, O E; Khudiakova, L V

2010-01-01

63

COPI Is Required for Enterovirus 71 Replication  

PubMed Central

Enterovirus 71 (EV71), a member of the Picornaviridae family, is found in Asian countries where it causes a wide range of human diseases. No effective therapy is available for the treatment of these infections. Picornaviruses undergo RNA replication in association with membranes of infected cells. COPI and COPII have been shown to be involved in the formation of picornavirus-induced vesicles. Replication of several picornaviruses, including poliovirus and Echovirus 11 (EV11), is dependent on COPI or COPII. Here, we report that COPI, but not COPII, is required for EV71 replication. Replication of EV71 was inhibited by brefeldin A and golgicide A, inhibitors of COPI activity. Furthermore, we found EV71 2C protein interacted with COPI subunits by co-immunoprecipitation and GST pull-down assay, indicating that COPI coatomer might be directed to the viral replication complex through viral 2C protein. Additionally, because the pathway is conserved among different species of enteroviruses, it may represent a novel target for antiviral therapies. PMID:22662263

Wang, Jianmin; Wu, Zhiqiang; Jin, Qi

2012-01-01

64

Enterovirus and herpesviridae family as etiologic agents of lymphomonocytary meningitis, Southern Brazil.  

PubMed

Viral meningitis is a common infectious disease of the central nervous system (CNS) that occurs worldwide. The aim of this study was to identify the etiologic agent of lymphomonocytary meningitis in Curitiba, PR, Brazil. During the period of July 2005 to December 2006, 460 cerebrospinal fluid (CSF) samples with lymphomonocytary meningitis were analyzed by PCR methodologies. Fifty nine (12.8%) samples were positive. Enteroviruses was present in 49 (83%) samples and herpes virus family in 10 (17%), of these 6 (10%) herpes simplex virus, 1 (2%) Epstein Barr virus, 2 (3%) human herpes virus type 6 and 1 (2%) mixed infection of enterovirus and Epstein Barr virus. As conclusion enterovirus was the most frequent virus, with circulation during summer and was observed with higher frequency between 4 to 17 years of age. PCR methodology is an important method for rapid detection of RNA enterovirus and DNA herpesvirus in CSF. PMID:21755125

Vidal, Luine Rosele Renaud; Almeida, Srgio Monteiro de; Messias-Reason, Iara Jos de; Nogueira, Meri Bordignon; Debur, Maria do Carmo; Pessa, Lus Felipe Cavalli; Pereira, Luciane Aparecida; Rotta, Indianara; Takahashi, Gislene Reche de Almeida; Silveira, Clyete Santos da; Arajo, Josianne Maria Reimann; Raboni, Sonia Mara

2011-06-01

65

Conformational Plasticity of the 2A Proteinase from Enterovirus 71  

PubMed Central

The 2A proteinase (2Apro) is an enterovirally encoded cysteine protease that plays essential roles in both the processing of viral precursor polyprotein and the hijacking of host cell translation and other processes in the virus life cycle. Crystallographic studies of 2Apro from enterovirus 71 (EV71) and its interaction with the substrate are reported here. EV71 2Apro was comprised of an N-terminal domain of a four-stranded antiparallel ? sheet and a C-terminal domain of a six-stranded antiparallel ? barrel with a tightly bound zinc atom. Unlike in other 2Apro structures, there is an open cleft across the surface of the protein in an open conformation. As demonstrated by the crystallographic studies and modeling of the complex structure, the open cleft could be fitted with the substrate. On comparison 2Apro of EV71 to those of the human rhinovirus 2 and coxsackievirus B4, the open conformation could be closed with a hinge motion in the bII2 and cII ? strands. This was supported by molecular dynamic simulation. The structural variation among different 2Apro structures indicates a conformational flexibility in the substrate-binding cleft. The open structure provides an accessible framework for the design and development of therapeutics against the viral target. PMID:23616646

Cai, Qixu; Yameen, Muhammad; Liu, Weihua; Gao, Zhenting; Li, Yaozong; Peng, Xuanjia; Cai, Yaxian; Wu, Caiming; Zheng, Qian

2013-01-01

66

[Fruit of the emergence of an enterovirus: acute haemorrhagic conjunctivitis].  

PubMed

First seen in Ghana and Indonesia in the early 70's, acute haemorrhagic conjunctivitis or "Apollo 11" disease is an eye infection caused by Enterovirus type 70 (EV70). The disease appeared to be a highly contagious conjunctivitis which spread rapidly all over the world. EV70 has been considered as an emerging virus and was classified as a new Enterovirus. No human or animal virus genetically similar to EV70 was known before the sudden outcome of the disease in Ghana, West Africa. EV70 appeared as a pretty demonstrative example of virus emergence and virus spreading. Studies of virus genetic mutations emphasized the variations of RNA virus within a short time period. The current review presents the EV70 infection and the genetic profile of the virus from its emergence to nowadays. PMID:18957336

Sane, F; Sauter, P; Fronval, S; Goffard, A; Dewilde, A; Hober, D

2008-01-01

67

112 The Development of Enterovirus 71 Vaccine  

PubMed Central

Enterovirus 71 (EV71) is a major cause of epidemic outbreaks of hand-foot-and-mouth disease (HFMD) worldwide. The virus belongs to the family Picornaviridae, genus Enterovirus. 3 genotypesA, B, and C and more than 10 sub-genotypes have been identified. After the initial identification of this virus in 1969, some large outbreaks of HFMD have been reported worldwide. EV71-induced HFMD is usually characterized by the formation of maculopapular or vesicular lesions on the skin and oral mucosa, especially on the palms, soles, and mouth. There were a greater number of fatal cases with brainstem encephalitis, pulmonary edema and/or hemorrhage, and cardiopulmonary collapse. Developing effective vaccines is considered a top choice among all control measures. We evaluated the ability of inactivated virus vaccine to elicit neutralizing antibody and to provide protection against lethal infection of EV71 in suckling mice. The purity of EV71 vaccine was up to 96.8% by HPLC identification. The purified EV71 vaccine induced high levels of neutralizing antibodies, these antibodies were shown to be protective against lethal infection when passively transferred to susceptible newborn mice. With a challenge dose of 50LD50 virus/mouse, suckling mice born to dams immunized with inactivated virus showed 100% survival. In preliminary animal trial, no side effects were detected when monkeys were immunized with purified EV71 vaccine either at normal or large doses. The vaccine was approved of the clinical evaluation in 2009. The phase 1/2/3 clinical trail was completed in March 2013. The data suggested that the inactivated EV71 vaccine had a clinically acceptable safety profile and good immunogenicity in healthy children and infants. Our data indicated that inactivated EV71 vaccine is the choice of vaccine preparation capable of fulfilling the demand for effective control.

Xiuling, Li; Zhongyang, Zhang; Xiaoxiao, Wang; Chunsheng, Hao; Yi, Li; Chong, Zhang; Chen, Zhang; Ming, Chen; haijing, Ning; Yu, Liu; Lei, Chen; Shuhua, Ma; Yongjuan, Yang; Dongmei, Song; Xinliang, Shen

2014-01-01

68

Human Vitamin B6 Requirements.  

National Technical Information Service (NTIS)

This is a report of a proceedings on human vitamin B6 requirements. Topics covered include: Vitamin B6 - chemistry, absorption, metabolism; vitamin B6 relationship in tryptophan metabolism; vitamin B6 and the metabolism of sulfur amino acids; analysis of ...

1978-01-01

69

High Susceptibility for Enterovirus Infection and Virus Excretion Features in Tunisian Patients with Primary Immunodeficiencies  

PubMed Central

To estimate the susceptibility to enterovirus infection and the frequency of long-term poliovirus excreters in Tunisian patients with primary immunodeficiencies (PIDs), enteroviruses were assessed in stool specimens of 82 patients with humoral, combined, and other PIDs. Isolated viruses were typed and intratyped by standard molecular techniques, and the whole VP1 region of poliovirus isolates was sequenced. Polioviruses were detected in 6 patients; all isolates were vaccine related. Five patients rapidly stopped excretion; one excreted a poliovirus type 1 isolate for several months, and the isolate accumulated up to 14 mutations in the VP1 region. Nonpolio enteroviruses were identified in 6 patients; 4 of them kept excreting the same strain for more than 6 months. The rate of enterovirus infection was 13.4% of the PID patients and 20.7% of those with an IgG defect; it greatly exceeded the rates generally found in Tunisian supposed-immunocompetent individuals (4.1% during the study period; P = 0.001 and P < 0.0001, respectively). Interestingly, patients with combined immunodeficiencies were at a higher risk for enterovirus infection than those with an exclusively B cell defect. A major histocompatibility complex (MHC) class II antigen expression defect was found in 54% of enterovirus-positive patients and in the unique long-term poliovirus excreter. The study results also suggest that substitutive immunoglobulin therapy may help clearance of a poliovirus infection and that most PID patients have the ability to stop poliovirus excretion within a limited period. However, the high susceptibility of these patients to enterovirus infection reinforces the need for enhanced surveillance of these patients until the use of oral poliovirus vaccine (OPV) is stopped. PMID:22914367

Driss, Nadia; Ben-Mustapha, Imen; Mellouli, Fethi; Ben Yahia, Ahlem; Touzi, Henda; Bejaoui, Mohamed; Ben Ghorbel, Mohamed; Barbouche, Mohamed-Ridha

2012-01-01

70

Transgenic mouse model for the study of enterovirus 71 neuropathogenesis  

PubMed Central

Enterovirus 71 (EV71) typically causes mild hand-foot-and-mouth disease in children, but it can also cause severe neurological disease. Recently, epidemic outbreaks of EV71 with significant mortality have been reported in the Asia-Pacific region, and EV71 infection has become a serious public health concern worldwide. However, there is little information available concerning EV71 neuropathogenesis, and no vaccines or anti-EV71 drugs have been developed. Previous studies of this disease have used monkeys and neonatal mice that are susceptible to some EV71 strains as models. The monkey model is problematic for ethical and economical reasons, and mice that are more than a few weeks old lose their susceptibility to EV71. Thus, the development of an appropriate small animal model would greatly contribute to the study of this disease. Mice lack EV71 susceptibility due to the absence of a receptor for this virus. Previously, we identified the human scavenger receptor class B, member 2 (hSCARB2) as a cellular receptor for EV71. In the current study, we generated a transgenic (Tg) mouse expressing hSCARB2 with an expression profile similar to that in humans. Tg mice infected with EV71 exhibited ataxia, paralysis, and death. The most severely affected cells were neurons in the spinal cord, brainstem, cerebellum, hypothalamus, thalamus, and cerebrum. The pathological features in these Tg mice were generally similar to those of EV71 encephalomyelitis in humans and experimentally infected monkeys. These results suggest that this Tg mouse could represent a useful animal model for the study of EV71 infection. PMID:23959904

Fujii, Ken; Nagata, Noriyo; Sato, Yuko; Ong, Kien Chai; Wong, Kum Thong; Yamayoshi, Seiya; Shimanuki, Midori; Shitara, Hiroshi; Taya, Choji; Koike, Satoshi

2013-01-01

71

Bovine enteroviruses: classification and serological characterization  

E-print Network

BOVINE ENTEROVIRUSES: CLASS IF ICATION AND SEROLOGICAL CHARACTERIZATION A Thesis by SIBA KUMAR SAMAL Submitted to the Graduate College of Texas A&M Uni vers i ty in partial fulfillment of the requirement for the degree of MASTER OF SCIENCE... August 1981 Major Subject: Veterinary Microbiology BOVINE ENTEROVIRUSES: CLASSIFICATION AND SEROLOGICAL CHARACTERIZATION A Thesis by SIBA KUMAR SAMAL Approved as to style and content by: Chairman o Committee Hem er Nem e Head of Oepartment...

Samal, Siba Kumar

2012-06-07

72

Bovine enteroviruses as indicators of fecal contamination.  

PubMed

Surface waters frequently have been contaminated with human enteric viruses, and it is likely that animal enteric viruses have contaminated surface waters also. Bovine enteroviruses (BEV), found in cattle worldwide, usually cause asymptomatic infections and are excreted in the feces of infected animals in large numbers. In this study, the prevalence and genotype of BEV in a closed herd of cattle were evaluated and compared with BEV found in animals in the immediate environment and in environmental specimens. BEV was found in feces from 76% of cattle, 38% of white-tailed deer, and one of three Canada geese sharing the same pastures, as well as the water obtained from animal watering tanks, from the pasture, from streams running from the pasture to an adjacent river, and from the river, which emptied into the Chesapeake Bay. Furthermore, BEV was found in oysters collected from that river downstream from the farm. These findings suggest that BEV could be used as an indicator of fecal pollution originating from animals (cattle and/or deer). Partial sequence analysis of the viral genomes indicates that different viral variants coexist in the same area. The possibility of identifying the viral strains found in the animals and in the contaminated areas by sequencing the RNA genome, could provide a tool to find the origin of the contamination and should be useful for epidemiological and viral molecular evolution studies. PMID:12089028

Ley, Victoria; Higgins, James; Fayer, Ronald

2002-07-01

73

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

2013-01-01

74

ADSORPTION OF ENTEROVIRUSES TO SOIL CORES AND THEIR SUBSEQUENT ELUTION BY ARTIFICIAL RAINWATER  

EPA Science Inventory

The adsorption and elution of a variety of human enteroviruses in a highly permeable, sandy soil was studied by using cores (43 by 125 mm) collected from an operating recharge basin on Long Island. Viruses studied included field and reference strains of polioviruses types 1 and 3...

75

Management of Atrial Tachycardia in the Newborn With Enterovirus Myocarditis  

PubMed Central

Neonatal enterovirus myocarditis is a rare but serious infection that is often an underrecognized cause of cardiovascular collapse. Enterovirus myocarditis in patients with such collapse should be suspected when signs of congestive heart failure and tachyarrhythmia are present. The majority of reported electrical disturbances associated with enterovirus myocarditis are ventricular in origin, but the infection can present as atrial tachyarrhythmia. Atrial tachyarrhythmias associated with enterovirus myocarditis are difficult to manage because of their resistance to conventional antiarrhythmic therapy. We present 2 cases of neonates with atrial tachycardia associated with enterovirus myocarditis who responded to a combination of amiodarone and flecainide. PMID:22778684

Petroni, Daniel H.; Yang, Song G.; Kattash, Mudar M.; Snyder, Christopher S.

2012-01-01

76

Comparison of human parechovirus and enterovirus detection frequencies in cerebrospinal fluid samples collected over a 5-year period in edinburgh: HPeV type 3 identified as the most common picornavirus type.  

PubMed

Human enteroviruses (EVs) and more recently parechoviruses (HPeVs) have been identified as the principal viral causes of neonatal sepsis-like disease and meningitis. The relative frequencies of specific EV and HPeV types were determined over a 5-year surveillance period using highly sensitive EV and HPeV PCR assays for screening 4,168 cerebrospinal fluid (CSF) specimens collected from hospitalized individuals between 2005 and 2010 in Edinburgh. Positive CSF samples were typed by sequencing of VP1. From the 201 EV and 31 HPeV positive (uncultured) CSF samples on screening, a high proportion of available samples could be directly typed (176/182, 97%). Highest frequencies of EV infections occurred in young adults (n = 43; 8.6%) although a remarkably high proportion of positive samples (n = 98; 46%) were obtained from young infants (<3 months). HPeV infections were seen exclusively in children under the age of 3 months (31/1,105; 2.8%), and confined to spring on even-numbered years (22% in March 2006, 25% in April 2008, and 22% in March 2010). In contrast, EV infections were distributed widely across the years. Twenty different EV serotypes were detected; E9, E6, and CAV9 being found most frequently, whereas all but one HPeVs were type 3. Over this period, HPeV3 was identified as the most prevalent picornavirus type in CNS-related infections with similarly high incidences of EV infection frequencies in very young children. The highly sensitive virus typing methods applied in this study will assist further EV and HPeV screening of sepsis and meningitis cases as well as in future molecular epidemiological studies and population surveillance. PMID:21412796

Harvala, Heli; McLeish, Nigel; Kondracka, Jasmina; McIntyre, Chloe L; McWilliam Leitch, E Carol; Templeton, Kate; Simmonds, Peter

2011-05-01

77

Enteroviruses and Bacteriophages in Bathing Waters  

Microsoft Academic Search

A new procedure for detecting and counting enteroviruses based on the VIRADEN method applied to 10 liters of seawater was examined. It improved the efficiency of detection by taking into account both the number of positive isolations and numbers found with traditional methods. It was then used to quantify viruses in bathing waters. A number of bacterial indicators and bacteriophages

L. Moce-Llivina; Francisco Lucena; Juan Jofre

2005-01-01

78

Enteroviruses in water environment--a potential threat to public health.  

PubMed

Enteroviruses belong to the Picornaviridae family and are the smallest, nonenveloped viruses known to infect both humans and animals. The spread of enteroviral infections is mainly by the faecal-oral and oral-oral route, but also through direct contact with secretions from ophthalmic and dermal lesions. Water, food and soil contaminated by infected faeces are an exogenous infection source which creates many opportunities for the transfer of the infection, and cause an epidemic outbreak in a short period of time. Enteroviruses are being isolated from all types of water: ground, sea, sewage and fresh water environments but also--and what is the most important from the epidemiological point of view--drinking water. They are resilient organisms, able to withstand high concentrations of sodium chloride (NaCl) and large changes in temperature. These abilities allow the viruses to flourish in a water environment, their natural reservoir. The number of infections in temperate climate regions peak in summer months and early autumn. Detection of enteroviruses in the water environment is performed by virus isolation in cell cultures and the use of molecular techniques. Many researches conducted in different countries with the use of methods mentioned above, reveal widespread environmental contamination by enteroviruses. PMID:19061255

Rajtar, Barbara; Majek, Magdalena; Pola?ski, ?ukasz; Polz-Dacewicz, Ma?gorzata

2008-12-01

79

Optimization of a Combined Human Parechovirus-Enterovirus Real-Time Reverse Transcription-PCR Assay and Evaluation of a New Parechovirus 3-Specific Assay for Cerebrospinal Fluid Specimen Testing  

PubMed Central

Human parechoviruses (HPeVs), particularly type 3 (HPeV3), are known central nervous system (CNS) pathogens, causing serious infections in infants similar to those caused by enteroviruses (EVs). The primary aim of this study was to combine and validate HPeV and EV real-time reverse transcription-PCR (RT-PCR) detection assays with the best available RT-PCR reagents and conditions for parallel detection of HPeV and EV on a single platform. The secondary aim was to develop and validate a newly developed HPeV3-specific real-time RT-PCR assay. Five commercially available RT-PCR kits were evaluated with the pan-HPeV and EV assays in one-step and two-step RT-PCRs. Two-step RT-PCR with the AgPath ID RT-PCR (AGP) kit performed best for both pan-HPeV and EV assays. The pan-HPeV-specific assay performed best with the AGP kit in a one-step RT-PCR. Frozen aliquots of 145 (for HPeV, n = 70; for EV, n = 75) previously characterized cerebrospinal fluid (CSF) specimens were tested by EV-, pan-HPeV-, and HPeV3-specific (HPeV specimens only) assays. The pan-HPeV and EV assays demonstrated 100% analytical sensitivity and specificity compared to historic results, while the HPeV3-specific assay demonstrated 97% sensitivity and 100% specificity. We propose a real-time pan-HPeV, EV two-step RT-PCR algorithm for simultaneous detection of HPeV and EV from CSF specimens on a single platform. The HPeV3-specific one-step RT-PCR assay can be used as a rapid and cost-effective assay to detect and identify HPeV3 in pan-HPeV RT-PCR assay-positive CSF specimens. PMID:23175256

Selvaraju, Suresh B.; Nix, W. Allan; Oberste, M. Steven

2013-01-01

80

Enterovirus surveillance--United States, 2000-2001.  

PubMed

Enteroviruses are common viruses associated with diverse clinical manifestations ranging from mild febrile illness to severe and potentially fatal syndromes including aseptic meningitis, encephalitis, neonatal systemic enteroviral disease, and paralytic poliomyelitis. A total of 64 enterovirus serotypes are recognized, including 61 nonpolio enteroviruses. Individual serotypes have different temporal patterns of circulation and often are associated with different clinical manifestations. This report describes temporal trends in reported enterovirus infections in the United States during 2000-2001, including widespread activity of two serotypes (echoviruses 13 and 18) that previously were detected rarely. Monitoring of circulating enterovirus serotypes helped identify these two agents as the primary causes of aseptic meningitis outbreaks in the United States in 2001. Further improvements in timeliness of reporting and geographic representation of the system are needed to allow more complete surveillance for enteroviruses. PMID:12487527

2002-11-22

81

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

PubMed

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

2012-01-01

82

Discovery of a Bovine Enterovirus in Alpaca  

PubMed Central

A cytopathic virus was isolated using Madin-Darby bovine kidney (MDBK) cells from lung tissue of alpaca that died of a severe respiratory infection. To identify the virus, the infected cell culture supernatant was enriched for virus particles and a generic, PCR-based method was used to amplify potential viral sequences. Genomic sequence data of the alpaca isolate was obtained and compared with sequences of known viruses. The new alpaca virus sequence was most similar to recently designated Enterovirus species F, previously bovine enterovirus (BEVs), viruses that are globally prevalent in cattle, although they appear not to cause significant disease. Because bovine enteroviruses have not been previously reported in U.S. alpaca, we suspect that this type of infection is fairly rare, and in this case appeared not to spread beyond the original outbreak. The capsid sequence of the detected virus had greatest homology to Enterovirus F type 1 (indicating that the virus should be considered a member of serotype 1), but the virus had greater homology in 2A protease sequence to type 3, suggesting that it may have been a recombinant. Identifying pathogens that infect a new host species for the first time can be challenging. As the disease in a new host species may be quite different from that in the original or natural host, the pathogen may not be suspected based on the clinical presentation, delaying diagnosis. Although this virus replicated in MDBK cells, existing standard culture and molecular methods could not identify it. In this case, a highly sensitive generic PCR-based pathogen-detection method was used to identify this pathogen. PMID:23950875

McClenahan, Shasta D.; Scherba, Gail; Borst, Luke; Fredrickson, Richard L.; Krause, Philip R.; Uhlenhaut, Christine

2013-01-01

83

[Experience in the use of chemical compounds for the primary identification and differentiation of enteroviruses].  

PubMed

Data on the usage of chemical inhibitors nifan and belvtazid, which possess a selective and antienteroviral effect, in the primary identification of enteroviruses and their differentiation into polio- and non-poliomyelytic ones isolated from human clinical materials or the environment by using the cell culture are presented in the article. The method is recommended for the practical use by the virology service in the diagnostics of enteroviral infections and in the identification of cytopathic agents isolated from the enviroment. PMID:12688219

Amvros'eva, T V; Votiakov, V I; Andreeva, O T; Kazinets, O N; Bogush, Z F; Sharko, R M; Kvacheva, Z B

2003-02-01

84

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

2000-01-01

85

Identification of human metapneumovirus genotypes A and B from clinical specimens by reverse transcription loop-mediated isothermal amplification.  

PubMed

Human metapneumovirus (hMPV) has been recognized as an important pathogen for acute respiratory infections in children worldwide and classified into genotypes A and B. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay is a rapid diagnostic method for detecting nucleic acids with a single step under isothermal conditions in less than 1h. RT-LAMP targeting the M gene of hMPV was developed for detecting and identifying hMPV genotypes A and B. The detection limit of the genotype-specific hMPV RT-LAMP assay was 10 times greater than that of conventional reverse transcription polymerase chain reaction (RT-PCR). No cross-reactivity was found with respiratory syncytial virus, parainfluenza virus 1-3, adenovirus, human bocavirus, human rhinovirus, influenza virus A and B, human coronaviruses and enteroviruses. One hundred and fifteen clinical specimens were detected for hMPV genotypes A and B with RT-LAMP, RT-PCR and real-time SYBR PCR. Kappa coefficients showed that there was a good agreement among these three methods. Compared with RT-PCR and real-time SYBR PCR, the genotype-specific RT-LAMP showed better specificity, sensitivity and is more convenient to perform with reduced turn-around time. PMID:24269205

Song, Qinwei; Zhu, Runan; Sun, Yu; Zhao, Linqing; Wang, Fang; Deng, Jie; Qian, Yuan

2014-02-01

86

Enter at Your Own Risk: How Enteroviruses Navigate the Dangerous World of Pattern Recognition Receptor Signaling  

PubMed Central

Enteroviruses are the most common human viral pathogens worldwide. This genus of small, non-enveloped, single stranded RNA viruses includes coxsackievirus, rhinovirus, echovirus, and poliovirus species. Infection with these viruses can induce mild symptoms that resemble the common cold, but can also be associated with more severe syndromes such as poliomyelitis, neurological diseases including aseptic meningitis and encephalitis, myocarditis, and the onset of type I diabetes. In humans, polarized epithelial cells lining the respiratory and/or digestive tracts represent the initial sites of infection by enteroviruses. Control of infection in the host is initiated through the engagement of a variety of pattern recognition receptors (PRRs). PRRs act as the sentinels of the innate immune system and serve to alert the host to the presence of a viral invader. This review assembles the available data annotating the role of PRRs in the response to enteroviral infection as well as the myriad ways by which enteroviruses both interrupt and manipulate PRR signaling to enhance their own replication, thereby inducing human disease. PMID:23764548

Harris, Katharine G; Coyne, Carolyn B

2013-01-01

87

Animal models of enterovirus 71 infection: applications and limitations  

PubMed Central

Human enterovirus 71 (EV71) has emerged as a neuroinvasive virus that is responsible for several outbreaks in the Asia-Pacific region over the past 15years. Appropriate animal models are needed to understand EV71 neuropathogenesis better and to facilitate the development of effective vaccines and drugs. Non-human primate models have been used to characterize and evaluate the neurovirulence of EV71 after the early outbreaks in late 1990s. However, these models were not suitable for assessing the neurovirulence level of the virus and were associated with ethical and economic difficulties in terms of broad application. Several strategies have been applied to develop mouse models of EV71 infection, including strategies that employ virus adaption and immunodeficient hosts. Although these mouse models do not closely mimic human disease, they have been applied to determine the pathogenesis of and treatment and prevention of the disease. EV71 receptor-transgenic mouse models have recently been developed and have significantly advanced our understanding of the biological features of the virus and the host-parasite interactions. Overall, each of these models has advantages and disadvantages, and these models are differentially suited for studies of EV71 pathogenesis and/or the pre-clinical testing of antiviral drugs and vaccines. In this paper, we review the characteristics, applications and limitation of these EV71 animal models, including non-human primate and mouse models. PMID:24742252

2014-01-01

88

Animal models of enterovirus 71 infection: applications and limitations.  

PubMed

Human enterovirus 71 (EV71) has emerged as a neuroinvasive virus that is responsible for several outbreaks in the Asia-Pacific region over the past 15 years. Appropriate animal models are needed to understand EV71 neuropathogenesis better and to facilitate the development of effective vaccines and drugs. Non-human primate models have been used to characterize and evaluate the neurovirulence of EV71 after the early outbreaks in late 1990s. However, these models were not suitable for assessing the neurovirulence level of the virus and were associated with ethical and economic difficulties in terms of broad application. Several strategies have been applied to develop mouse models of EV71 infection, including strategies that employ virus adaption and immunodeficient hosts. Although these mouse models do not closely mimic human disease, they have been applied to determine the pathogenesis of and treatment and prevention of the disease. EV71 receptor-transgenic mouse models have recently been developed and have significantly advanced our understanding of the biological features of the virus and the host-parasite interactions. Overall, each of these models has advantages and disadvantages, and these models are differentially suited for studies of EV71 pathogenesis and/or the pre-clinical testing of antiviral drugs and vaccines. In this paper, we review the characteristics, applications and limitation of these EV71 animal models, including non-human primate and mouse models. PMID:24742252

Wang, Ya-Fang; Yu, Chun-Keung

2014-01-01

89

Cell and tissue tropism of enterovirus 71 and other enteroviruses infections.  

PubMed

Enterovirus 71 (EV71) is a member of Picornaviridae that causes mild and self-limiting hand, foot, and mouth disease (HFMD). However, EV71 infections can progress to polio-like paralysis, neurogenic pulmonary edema, and fatal encephalitis in infants and young children. Large EV71 outbreaks have been reported in Taiwan, China, Japan, Malaysia, Singapore, and Australia. This virus is considered a critical emerging public health threat. EV71 is an important crucial neurotropic enterovirus for which there is currently no effective antiviral drug or vaccine. The mechanism by which EV71 causes severe central nervous system complications remains unclear. The interaction between the virus and the host is vital for viral replication, virulence, and pathogenicity. SCARB2 or PSGL-1 receptor binding is the first step in the development of viral infections, and viral factors (e.g., 5' UTR, VP1, 3C, 3D, 3' UTR), host factors and environments (e.g., ITAFs, type I IFN) are also involved in viral infections. The tissue tropism and pathogenesis of viruses are determined by a combination of several factors. This review article provides a summary of host and virus factors affecting cell and tissue tropism and the pathogenesis of enteroviruses. PMID:24602216

Lin, Jing-Yi; Shih, Shin-Ru

2014-01-01

90

Clinical severity of rhinovirus/enterovirus compared to other respiratory viruses in children  

PubMed Central

Background Human rhinovirus/enterovirus (HRV/ENT) infections are commonly identified in children with acute respiratory infections (ARIs), but data on their clinical severity remain limited. Objectives We compared the clinical severity of HRV/ENT to respiratory syncytial virus (RSV), influenza A/B (FLU), and other common respiratory viruses in children. Patients/Methods Retrospective study of children with ARIs and confirmed single positive viral infections on mid-turbinate swabs by molecular assays. Outcome measures included hospital admission and, for inpatients, a composite endpoint consisting of intensive care admission, hospitalization >5 days, oxygen requirements or death. Results A total of 116 HRV/ENT, 102 RSV, 99 FLU, and 64 other common respiratory viruses were identified. Children with single HRV/ENT infections presented with significantly higher rates of underlying immunosuppressive conditions compared to those with RSV (379% versus 136%; P < 0001), FLU (379% versus 22%; P = 0018) or any other single viral infection (379% versus 225%; P = 0024). In multivariable analysis adjusted for underlying conditions and age, children with HRV/ENT infections had increased odds of hospitalization compared to children with RSV infections (OR 26; 95% CI 14, 48; P < 0003) or FLU infections (OR 30; 95% CI 16, 58; <0001) and increased odds of severe clinical disease among inpatients (OR 30; 95% CI 16,56; P = 0001) when compared to those with FLU infections. Conclusions Children with HRV/ENT had a more severe clinical course than those with RSV and FLUA/B infections and often had significant comorbidities. These findings emphasize the importance of considering HRV/ENT infection in children presenting with severe acute respiratory tract infections. PMID:24801963

Asner, Sandra A; Petrich, Astrid; Hamid, Jemila S; Mertz, Dominik; Richardson, Susan E; Smieja, Marek

2014-01-01

91

Enterovirus infection in Korean children and anti-enteroviral potential candidate agents  

PubMed Central

Although most enterovirus infections are not serious enough to be life threatening, several enteroviruses such as enterovirus 71 are responsible for severe, potentially life-threatening disease. The epidemic patterns of enteroviruses occur regularly during the year, but they may change due to environmental shifts induced by climate change due to global warming. Therefore, enterovirus epidemiological studies should be performed continuously as a basis for anti-viral studies. A great number of synthesized antiviral compounds that work against enteroviruses have been developed but only a few have demonstrated effectiveness in vivo. No proven effective antiviral agents are available for enterovirus disease therapy. The development of a new antiviral drug is a difficult task due to poor selective toxicity and cost. To overcome these limitations, one approach is to accelerate the availability of other existing antiviral drugs approved for antiviral effect against enteroviruses, and the other way is to screen traditional medicinal plants. PMID:23133481

Park, Kwi Sung; Choi, Young Jin

2012-01-01

92

Viral Pollution in the Environment and in Shellfish: Human Adenovirus Detection by PCR as an Index of Human Viruses  

PubMed Central

A study of the presence of human viruses (adenoviruses, enteroviruses, and hepatitis A viruses [HAVs]) in environmental and shellfish samples was carried out by applying DNA and cDNA amplification techniques by PCR. The detection of human adenoviruses by PCR was also examined as a potential molecular test to monitor viral pollution. The samples studied were urban and slaughterhouse sewage, river water, seawater, and shellfish. Enteroviruses were quantified by PFU in Buffalo green monkey kidney cells and fecal coliforms and phages of Bacteroides fragilis HSP40 were also evaluated in some of the samples. The amplification of viral DNA and cDNA has shown a high prevalence of human viruses that would not be detected by the use of classical techniques, such as the quantification of PFU in cell lines. The results of the analysis of slaughterhouse sewage samples together with the test of farm animal feces indicate that the adenoviruses and the HAVs detected in the environment are mostly of human origin. A significative correlation between the detection of human viruses by PCR and the values of bacteriophages of B. fragilis HSP40 in urban raw sewage was observed. Human adenoviruses were the viruses most frequently detected throughout the year, and all the samples that were positive for enteroviruses or HAVs were also positive for human adenoviruses. The results suggest that the detection of adenoviruses by PCR could be used as an index of the presence of human viruses in the environment where a molecular index is acceptable. PMID:9726885

Pina, Sonia; Puig, Montserrat; Lucena, Francisco; Jofre, Joan; Girones, Rosina

1998-01-01

93

Seroepidemiology and Molecular Epidemiology of Enterovirus 71 in Russia  

PubMed Central

Enterovirus 71 (EV71) is an emerging human pathogen causing massive epidemics of hand, foot and mouth disease with severe neurological complications in Asia. EV71 also circulates in Europe, however it does not cause large outbreaks. The reason for distinct epidemiological patterns of EV71 infection in Europe and Asia and the risk of EV71 epidemic in Europe and Russia remain unknown. Seroepidemiology of EV71 and molecular epidemiology of occasional EV71 isolates were studied to explore circulation of EV71 in Russia. In six regions of Russian Federation, seroprevalence of EV71 in sera collected in 2008 ranged from 5% to 20% in children aged 12 years and from 19% to 83% in children aged 35 years. The seroprevalence among elder children was significantly higher (4183% vs. 1927%) in Asian regions of Russia. EV71 strains identified in Russia in 20012011 belonged to subtypes C1 and C2, while genotype C4 that was causing epidemics in Asia since 1998 emerged in 2009 and became dominant in 2013. PMID:24819617

Akhmadishina, Ludmila V.; Eremeeva, Tatiana P.; Trotsenko, Olga E.; Ivanova, Olga E.; Mikhailov, Mikhail I.; Lukashev, Alexander N.

2014-01-01

94

Molecular epidemiology of enterovirus 71 strains isolated from children in Yamagata, Japan, between 1990 and 2013.  

PubMed

Enterovirus 71 infections have become a major public issue in the Asia-Pacific region due to the large number of fatal cases. To clarify the longitudinal molecular epidemiology of enterovirus 71 (EV71) in a community, we isolated 240 strains from children, mainly with hand-foot-and-mouth diseases, between 1990 and 2013 in Yamagata, Japan. We carried out a sequence analysis of the VP1 region (891 bp) using 223 isolates and identified six subgenogroups (B2, B4, B5, C1, C2 and C4) during the study period. Subgenogroups C1 and B2 were found only between 1990 and 1993 and have not reappeared since. In contrast, strains in subgenogroups C2, C4 and B5 appeared repeatedly with genomic variations. Recent reports from several local communities in Japan have suggested that identical predominant subgenogroup strains, which have also been found in the Asia-Pacific region, have been circulating in a wide area in Japan. However, it is likely that there is a discrepancy between the major subgenogroups circulating in the Asia-Pacific region and those in Europe. It is necessary to continue the analysis of the longitudinal epidemiology of EV71 in local communities, as well as on regional and global levels, to develop strategies against severe EV71 infections. PMID:25053796

Mizuta, Katsumi; Aoki, Yoko; Matoba, Yohei; Yahagi, Kazue; Itagaki, Tsutomu; Katsushima, Fumio; Katsushima, Yuriko; Ito, Sueshi; Hongo, Seiji; Matsuzaki, Yoko

2014-10-01

95

High Susceptibility for Enterovirus Infection and Virus Excretion Features in Tunisian Patients with Primary Immunodeficiencies  

E-print Network

High Susceptibility for Enterovirus Infection and Virus Excretion Features in Tunisian Patients, Ministry of Health, Tunis, Tunisiad To estimate the susceptibility to enterovirus infection), enteroviruses were assessed in stool specimens of 82 patients with humoral, com- bined, and other PIDs. Isolated

Paris-Sud XI, Université de

96

Co-Circulation and Evolution of Polioviruses and Species C Enteroviruses in a District  

E-print Network

Co-Circulation and Evolution of Polioviruses and Species C Enteroviruses in a District and phenotypic characteristics and compared them with co-circulating enteroviruses. These VDPVs appeared poliovaccine (OPV) in the 59-half of the genome and sequences derived from unidentified species C enteroviruses

Paris-Sud XI, Université de

97

Characterization of the Non-Polio Enterovirus Infections Associated with Acute Flaccid Paralysis in South-Western India  

PubMed Central

Non-polio enteroviruses (NPEVs) have been reported frequently in association with acute flaccid paralysis (AFP) cases during Polio Surveillance Programs (PSPs) worldwide. However, there is limited understanding on the attributes of their infections. This study reports characteristics of NPEVs isolated from AFP cases, investigated during PSPs held in 20092010, in Karnataka and Kerala states of south-western India having varied climatic conditions. NPEV cell culture isolates derived from stool specimens that were collected from 422 of 2186 AFP cases (<114 years age) and 17 of 41 asymptomatic contacts; and details of all AFP cases/contacts were obtained from National Polio Laboratory, Bangalore. The distribution of NPEV infections among AFP cases and circulation pattern of NPEV strains were determined by statistical analysis of the data. Genotyping of all NPEV isolates was carried out by partial VP1 gene sequencing and phylogenetic analysis. NPEV positive AFP cases were significantly higher in children aged <2 years; with residual paralysis; in summer months; and in regions with relatively hot climate. Genotyping of NPEVs identified predominance of human enteroviruses (HEV)-B species [81.9%Echoviruses (E): 57.3%; coxsackieviruses (CV) B: 15%; numbered EVs: 8.9%; CVA9: 0.7%] and low levels of HEV-A [14.5%CVA: 6%; numbered EVs: 8.5%] and HEV-C [3.6%CVA: 2.6%; numbered EVs: 1%] species, encompassing 63 genotypes. EV76 (6.3%) and each of E3, CVB3 and E9 (4.97%) were found frequently during 2009 while E11 (6.7%), CVB1 (6.1%), E7 (5.1%) and E20 (5.1%) were detected commonly in 2010. A marked proportion of AFP cases from children aged <2 years; presenting with fever; and from north and south interior parts of Karnataka state was detected with E/numbered EVs than that found with CVA/CVB. This study highlights the extensive genetic diversity and diverse circulation patterns of NPEV strains in AFP cases from different populations and climatic conditions. PMID:23630606

Laxmivandana, Rongala; Yergolkar, Prasanna; Gopalkrishna, Varanasi; Chitambar, Shobha D.

2013-01-01

98

Autoprocessing: an essential step for expression and purification of enterovirus 71 3C(pro) in Escherichia coli.  

PubMed

A gene encoding the 3BC of human enterovirus 71 (EV71) was cloned and inserted into a derivative of plasmid pET-32a(+) driven by T7 promoter. The expressed 3C protease (3C(pro)) autocatalytically cleaved itself from the recombinant protein Trx-3BC and the mature 3C(pro) partitioned in the soluble fraction of bacterial lysate. The 13-amino-acid peptide substrates with the junction of 3B/3C were used to verify the proteolysis activity of the purified 3C(pro). The EV71 3C(pro) had a Km value of 63 ?M (measured by a continuous fluorescence assay). The other solid-phase activity assay of the EV71 3C(pro) was developed using HPLC to analyze the proteolytic products. The combination of two activity assays contributes to promote the identification of the specific inhibitors targeted to the EV71 3C(pro). PMID:23881322

Huang, Shuqiong; Lyu, Yanning; Qing, Xianyun; Wang, Weiwei; Tang, Liang; Cheng, Kedi; Wang, Wei

2013-11-01

99

Comparison of diagnostic clinical samples and environmental sampling for enterovirus and parechovirus surveillance in Scotland, 2010 to 2012.  

PubMed

Human enteroviruses (EV) and parechoviruses (HPeV) within the family Picornaviridae are the most common causes of viral central nervous system (CNS)-associated infections including meningitis and neonatal sepsis-like disease. The frequencies of EV and HPeV types identified in clinical specimens collected in Scotland over an eight-year period were compared to those identified in sewage surveillance established in Edinburgh. Of the 35 different EV types belonging to four EV species (A to D) and the four HPeV types detected in this study, HPeV3 was identified as the most prevalent picornavirus in cerebrospinal fluid samples, followed by species B EV. Interestingly, over half of EV and all HPeV CNS-associated infections were observed in young infants (younger than three months). Detection of species A EV including coxsackievirus A6 and EV71 in clinical samples and sewage indicates that these viruses are already widely circulating in Scotland. Furthermore, species C EV were frequently identified EV in sewage screening but they were not present in any of 606 EV-positive clinical samples studied, indicating their likely lower pathogenicity. Picornavirus surveillance is important not only for monitoring the changing epidemiology of these infections but also for the rapid identification of spread of emerging EV and/or HPeV types. PMID:24762664

Harvala, H; Calvert, J; Van Nguyen, D; Clasper, L; Gadsby, N; Molyneaux, P; Templeton, K; McWilliams Leitch, C; Simmonds, P

2014-01-01

100

Characterization and specificity of the linear epitope of the enterovirus 71 VP2 protein  

PubMed Central

Background Enterovirus 71 (EV71) has emerged as a major causative agent of hand, foot and mouth disease in the Asia-Pacific region over the last decade. Hand, foot and mouth disease can be caused by different etiological agents from the enterovirus family, mainly EV71 and coxsackieviruses, which are genetically closely related. Nevertheless, infection with EV71 may occasionally lead to high fever, neurologic complications and the emergence of a rapidly fatal syndrome of pulmonary edema associated with brainstem encephalitis. The rapid progression and high mortality of severe EV71 infection has highlighted the need for EV71-specific diagnostic and therapeutic tools. Monoclonal antibodies are urgently needed to specifically detect EV71 antigens from patient specimens early in the infection process. Furthermore, the elucidation of viral epitopes will contribute to the development of targeted therapeutics and vaccines. Results We have identified the monoclonal antibody 7C7 from a screen of hybridoma cells derived from mice immunized with the EV71-B5 strain. The linear epitope of 7C7 was mapped to amino acids 142-146 (EDSHP) of the VP2 capsid protein and was characterized in detail. Mutational analysis of the epitope showed that the aspartic acid to asparagine mutation of the EV71 subgenogroup A (BrCr strain) did not interfere with antibody recognition. In contrast, the serine to threonine mutation at position 144 of VP2, present in recently emerged EV71-C4 China strains, abolished antigenicity. Mice injected with this virus strain did not produce any antibodies against the VP2 protein. Immunofluorescence and Western blotting confirmed that 7C7 specifically recognized EV71 subgenogroups and did not cross-react to Coxsackieviruses 4, 6, 10, and 16. 7C7 was successfully used as a detection antibody in an antigen-capture ELISA assay. Conclusions Detailed mapping showed that the VP2 protein of Enterovirus 71 contains a single, linear, non-neutralizing epitope, spanning amino acids 142-146 which are located in the VP2 protein's E-F loop. The S/T(144) mutation in this epitope confers a loss of VP2 antigenicity to some newly emerged EV71-C4 strains from China. The corresponding monoclonal antibody 7C7 was used successfully in an AC-ELISA and did not cross-react to coxsackieviruses 4, 6, 10, and 16 in immunofluorescence assay and Western blots. 7C7 is the first monoclonal antibody described, that can differentiate Coxsackievirus 16 from Enterovirus 71. PMID:22361222

2012-01-01

101

Cathepsin B expression in human tumors.  

PubMed

Cathepsin B has been linked to tumor progression through observations that its activity, secretion or membrane association are increased. The most malignant tumors, and specifically the cells at the invasive edge of those tumors, express the highest activity. Cathepsin B may facilitate invasion directly by dissolving extracellular matrix barriers like the basement membrane, or indirectly by activating other proteases capable of digesting the extracellular matrix. Cathepsin B also might play a role in tumor growth and angiogenesis. Cathepsin B activity is the result of several levels of regulation: transcription, post-transcription processing, translation and glycosylation, maturation and trafficking, and inhibition. The majority of reports on cathepsin B expression in tumors have focused on measurements of activity or protein staining. In some tumors, e.g. gliomas, a correlation between the amounts of cathepsin B mRNA, protein and activity and tumor progression has been established. Regulation of cathepsin B at the transcriptional and post-transcriptional levels is still poorly understood. Although the putative promoter regions have characteristics of housekeeping-type promoters, cathepsin B mRNA expression varies depending on the cell type and state of differentiation. We have evidence that more than one promoter could direct expression of human cathepsin B. Multiple transcript species have been detected, resulting from alternative splicing in the 5'- or 3'-untranslated regions, and possibly the use of alternative promoter regions. The existence of transcript variants indicates a potential for post-transcriptional control of expression. In support of this, ras-transformation of MCF-10A human breast epithelial cells results in an increase in protein levels without a concomitant increase in mRNA levels. Cathepsin B mRNA species with distinct 5'- or 3'-untranslated regions may differ in their stability and translatability. Variations in the coding region may also alter cathepsin B properties. We and Frankfater's group have observed transcript species that would encode a truncated protein, lacking the prepeptide and about half of the propeptide. This truncated protein, if synthesized in cells, would be expected to be cytosolic; therefore its function is unclear. Once the several mechanisms of regulation of cathepsin B expression and activity are better understood, they could provide us with new strategies to specifically reduce cathepsin B activity in tumors. PMID:8861022

Berquin, I M; Sloane, B F

1996-01-01

102

Characterization of a human B cell-specific antigen (B2) distinct from B1.  

PubMed

A human B lymphocyte-specific antigen (B2) was identified and characterized by the use of a monoclonal antibody. By indirect immunofluorescence and quantitative absorption, B2 was shown to be expressed exclusively on Ig+ B cells isolated from peripheral blood and lymphoid tissues. In contrast, B2 was not found on monocytes, resting and activated T cells, Null cells, or granulocytes, nor was it found on cell lines or tumor cells of T cell or myeloid origin. Functional studies demonstrated that only B2 antigen-positive splenocytes could be induced to differentiate into plasma cells under the stimulus of pokeweed mitogen, further confirming the B cell specificity of B2. It was then demonstrated that the B2 antigen was distinct from the previously described B cell-surface determinants including surface immunoglobulin, Ia-like antigens, and Fc and C3 receptors. More importantly, the B2 antigen has been clearly shown to be distinct from the previously described B cell-specific antigen, B1, by its m.w. and expression on normal and malignant B lymphocytes. The distinct distribution of B2 on normal and malignant lymphocytes supports the notion of B cell heterogeneity and provides further evidence for existence of subpopulations of human B lymphocytes. PMID:6163824

Nadler, L M; Stashenko, P; Hardy, R; van Agthoven, A; Terhorst, C; Schlossman, S F

1981-05-01

103

Recombination in the Evolution of Enterovirus C Species Sub-Group that Contains Types CVA-21, CVA-24, EV-C95, EV-C96 and EV-C99  

PubMed Central

Genetic recombination is considered to be a very frequent phenomenon among enteroviruses (Family Picornaviridae, Genus Enterovirus). However, the recombination patterns may differ between enterovirus species and between types within species. Enterovirus C (EV-C) species contains 21 types. In the capsid coding P1 region, the types of EV-C species cluster further into three sub-groups (designated here as AC). In this study, the recombination pattern of EV-C species sub-group B that contains types CVA-21, CVA-24, EV-C95, EV-C96 and EV-C99 was determined using partial 5?UTR and VP1 sequences of enterovirus strains isolated during poliovirus surveillance and previously published complete genome sequences. Several inter-typic recombination events were detected. Furthermore, the analyses suggested that inter-typic recombination events have occurred mainly within the distinct sub-groups of EV-C species. Only sporadic recombination events between EV-C species sub-group B and other EV-C sub-groups were detected. In addition, strict recombination barriers were inferred for CVA-21 genotype C and CVA-24 variant strains. These results suggest that the frequency of inter-typic recombinations, even within species, may depend on the phylogenetic position of the given viruses. PMID:24722726

Smura, Teemu; Blomqvist, Soile; Vuorinen, Tytti; Ivanova, Olga; Samoilovich, Elena; Al-Hello, Haider; Savolainen-Kopra, Carita; Hovi, Tapani; Roivainen, Merja

2014-01-01

104

A Retrospective Overview of Enterovirus Infection Diagnosis and Molecular Epidemiology in the Public Hospitals of Marseille, France (1985-2005)  

PubMed Central

Human enteroviruses (HEV) are frequent human pathogens and, associated in particular with large outbreaks of aseptic meningitis. Here, we have compiled a database of clinical HEV isolates from the Public Hospitals of Marseille, from 1985 to 2005. Amongst 654 isolates that could be characterized by complete sequencing of the VP1 gene, 98% belonged to species HEV-B; the most frequently isolated serotypes were Echovirus E30, E11, E7, E6 and E4. The high incidence of E30 and the recent emergence of E13 are consistent with reports worldwide and peak HEV isolation occurred mostly in the late spring and summer months. The proportion of echoviruses has decreased across the years, while that of coxsackieviruses has increased. Stool (the most frequent sample type) allowed detection of all identified serotypes. MRC5 (Human lung fibroblasts) cell line was the most conducive cell line for HEV isolation (84.9% of 10 most common serotype isolates, 96.3% in association with BGM (Buffalo green monkey kidney cells)). Previous seroneutralization-based serotype identification demonstrated 55.4% accuracy when compared with molecular VP1 analysis. Our analysis of a large number of clinical strains over 20 years reinforced the validity of VP1 serotyping and showed that comparative p-distance scores can be coupled with phylogenetic analysis to provide non-ambiguous serotype identification. Phylogenetic analysis in the VP1, 2C and 3D regions also provided evidence for recombination events amongst clinical isolates. In particular, it identified isolates with dissimilar VP1 but almost identical nonstructural regions. PMID:21437207

Tan, Charlene Y. Q.; Ninove, Laetitia; Gaudart, Jean; Nougairede, Antoine; Zandotti, Christine; Thirion-Perrier, Laurence; Charrel, Remi N.; de Lamballerie, Xavier

2011-01-01

105

Rapid One-Step Quantitative Reverse Transcriptase PCR Assay with Competitive Internal Positive Control for Detection of Enteroviruses in Environmental Samples  

Microsoft Academic Search

Human enteroviruses can serve as a more accurate indicator of human fecal contamination than conven- tional bacteriological fecal indicators. We describe here a quantitative reverse transcriptase PCR (qRT-PCR) assay specifically tailored to detect these viruses in environmental waters. The assay included a competitive internal positive control (CIPC) that allowed the inhibition of qRT-PCRs to be quantitatively assessed. Coamplification of the

Jason B. Gregory; R. Wayne Litaker; Rachel T. Noble

2006-01-01

106

Differential apoptosis gene expressions of rhabdomyosarcoma cells in response to enterovirus 71 infection  

PubMed Central

Background Enterovirus 71 (EV71) infection can induce the apoptosis of infected cells. The aim of this study is to explore the effect of EV71 infection on apoptosis mechanisms in virus-infected human rhabdomyosarcoma (RD) cells. Methods The apoptosis of RD cells was examined using annexin V-FITC/PI by flow cytometry and cytokines were detected by ELISA. Cellular RNA was extracted and transcribed to cDNA. PCR array was employed to analyze the expressions of 84 apoptotic genes from EV71-infected RD cells at 8 and 20 h postinfection, respectively. In addition, the expressions of FasL, caspase, AKT2, JNK1/2, c-Jun and NF-?B proteins were detected by western blotting. Results Flow cytometry demonstrated that the apoptosis or death of EV71-infected RD cells was increased by 37.1% with a multiplicity of infection (MOI) of 5 at 20 h postinfection. The production of IL-4, IL-10 and TNF-? was enhanced by the subsequent EV71 infection. PCR array revealed significant changes in the expressions of apoptotic genes. Among 84 genes, 42 genes were down-regulated after EV71 infection at 8 h, whereas 32 genes were up-regulated at 20 h postinfection. Moreover, the ligands of TNF superfamily such as FasL, CD40L and TNF-? were significantly up-regulated and enhanced the expressions of apoptosis-related cysteine peptidases, including caspase-10, -8, -7 and -3. In addition, EV71 infection induces the phosphorylation of AKT2, JNK1/2, c-Jun and NF-?B at 20 h postinfection. Conclusion PCR array for the determination of apoptosis gene expressions is an informative assay in elucidating biological pathways. During the early stage of EV71 infection, the apoptotic process of RD cells is significantly delayed. EV71 infection can also induce the expressions of FasL, TNF-? and CD40L, which contribute to the apoptosis of RD cells. PMID:23191987

2012-01-01

107

Sub-clinical enterovirus infections in Norwegian infants: A prospective cohort study on viral circulation and predictors of infection.  

E-print Network

??Background: Enteroviruses are common in infancy, but usually sub-clinical and self-limiting. Most previous data on enterovirus circulation derive from analyses of specimens from individuals with (more)

Wits, Elisabet

2009-01-01

108

RAG Mutations in Human B Cell-Negative SCID  

Microsoft Academic Search

Patients with human severe combined immunodeficiency (SCID) can be divided into those with B lymphocytes (B^+ SCID) and those without (B^- SCID). Although several genetic causes are known for B^+ SCID, the etiology of B^- SCID has not been defined. Six of 14 B^- SCID patients tested were found to carry a mutation of the recombinase activating gene 1 (RAG-1),

Klaus Schwarz; George H. Gauss; Leopold Ludwig; Ulrich Pannicke; Zhong Li; Doris Lindner; Wilhelm Friedrich; Reinhard A. Seger; Thomas E. Hansen-Hagge; Stephen Desiderio; Michael R. Lieber; Claus R. Bartram

1996-01-01

109

Functional Comparison of SCARB2 and PSGL1 as Receptors for Enterovirus 71  

PubMed Central

Human scavenger receptor class B, member 2 (SCARB2), and P-selectin glycoprotein ligand-1 (PSGL1) have been identified to be the cellular receptors for enterovirus 71 (EV71). We compared the EV71 infection efficiencies of mouse L cells that expressed SCARB2 (L-SCARB2) and PSGL1 (L-PSGL1) and the abilities of SCARB2 and PSGL1 to bind to the virus. L-SCARB2 cells bound a reduced amount of EV71 compared to L-PSGL1 cells. However, EV71 could infect L-SCARB2 cells more efficiently than L-PSGL1 cells. The results suggested that the difference in the binding capacities of the two receptors was not the sole determinant of the infection efficiency and that SCARB2 plays an essential role after attaching to virions. Therefore, we examined the viral entry into L-SCARB2 cells and L-PSGL1 cells by immunofluorescence microscopy. In both cells, we detected internalized EV71 virions that colocalized with an early endosome marker. We then performed a sucrose density gradient centrifugation analysis to evaluate viral uncoating. After incubating the EV71 virion with L-SCARB2 cells or soluble SCARB2 under acidic conditions below pH 6.0, we observed that part of the native virion was converted into an empty capsid that lacked both genomic RNA and VP4 capsid proteins. The results suggested that the uncoating of EV71 requires both SCARB2 and an acidic environment and occurs after the internalization of the virus-receptor complex into endosomes. However, the empty capsid formation was not observed after incubation with L-PSGL1 cells or soluble PSGL1 under any of the tested pH conditions. These results indicated that SCARB2 is capable of viral binding, viral internalization, and viral uncoating and that the low infection efficiency of L-PSGL1 cells is due to the inability of PSGL1 to induce viral uncoating. The characterization of SCARB2 as an uncoating receptor greatly contributes to the understanding of the early steps of EV71 infection. PMID:23302872

Yamayoshi, Seiya; Ohka, Seii; Fujii, Ken

2013-01-01

110

High Frequency and Diversity of Species C Enteroviruses in Cameroon and Neighboring Countries  

PubMed Central

Human enteroviruses (HEVs) are endemic worldwide and among the most common viruses infecting humans. Nevertheless, there are very limited data on the circulation and genetic diversity of HEVs in developing countries and sub-Saharan Africa in particular. We investigated the circulation and genetic diversity of HEVs among 436 healthy children in a limited area of the far north region of Cameroon in 2008 and 2009. We also characterized the genetic biodiversity of 146 nonpolio enterovirus (NPEV) isolates obtained throughout the year 2008 from stool specimens of patients with acute flaccid paralysis (AFP) in Cameroon, Chad, and Gabon. We found a high rate of NPEV infections (36.9%) among healthy children in the far north region of Cameroon. Overall, 45 different HEV types were found among healthy children and AFP patients. Interestingly, this study uncovered a high rate of HEVs of species C (HEV-C) among all typed NPEVs: 63.1% (94/149) and 39.5% (49/124) in healthy children and AFP cases, respectively. Besides extensive circulation, the most prevalent HEV-C type, coxsackievirus A-13, featured a tremendous intratypic diversity. Africa-specific HEV lineages were discovered, including HEV-C lineages and the recently reported EV-A71 genogroup E. Virtually all pathogenic circulating vaccine-derived polioviruses (cVDPVs) that have been fully characterized were recombinants between oral poliovaccine (OPV) strains and cocirculating HEV-C strains. The extensive circulation of diverse HEV-C types and lineages in countries where OPV is massively used constitutes a major viral factor that could promote the emergence of recombinant cVDPVs in the Central African subregion. PMID:23254123

Sadeuh-Mba, Serge Alain; Bessaud, Mael; Massenet, Denis; Joffret, Marie-Line; Endegue, Marie-Claire; Njouom, Richard; Reynes, Jean-Marc; Rousset, Dominique

2013-01-01

111

Human B cell defects in perspective.  

PubMed

While primary immune defects are generally considered to lead to severe and easily recognized disease in infants and children, a number of genetic defects impairing B cell function may not be clinically apparent or diagnosed until adult life. The commonest of these is common variable immune deficiency, the genetic origins of which are beginning to be at least partially understood. CVID affects ? 1/25,000 Caucasians and is characterized by a marked reduction in serum IgG, almost always in serum IgA, and reduced serum IgM in about half of all cases; these defects continue to provide an opportunity to investigate the genes necessary for B cell function in humans. Recently, a small number of genes necessary for normal B cell function have been identified in consanguineous families leading to varying degrees of hypogammaglobulinemia and loss of antibody production. In other studies, whole-exome sequencing and copy number variation, applied to large cohorts, have extended research into understanding both the genetic basis of this syndrome and the clinical phenotypes of CVID. PMID:22477523

Cunningham-Rundles, Charlotte

2012-12-01

112

Complete Genome Sequence of a Coxsackievirus B3 Isolated from a Sichuan Snub-Nosed Monkey  

PubMed Central

Coxsackievirus B3 (CVB3) is an enterovirus in the family Picornaviridae that is significant to human health, being associated with myocarditis, aseptic meningitis, and pancreatitis, among other conditions. In addition to humans, Sichuan snub-nosed monkeys can be infected and killed by CVB3. Here, we report the first complete genome sequence of a novel coxsackievirus B3 strain, SSM-CVB3, which was isolated from a deceased Sichuan snub-nosed monkey with severe myocarditis. Our findings may aid in understanding the evolutionary characteristics and molecular pathogenesis of this virus. PMID:23118460

He, Wenqi; Lu, Huijun; Zhao, Kui; Song, Deguang; Gai, Xianying

2012-01-01

113

Officials Report First Confirmed Child Death Due to Enterovirus D68  

MedlinePLUS

... Officials Report First Confirmed Child Death Due to Enterovirus D68 Virus, which can cause paralysis, has spread to ... 6, 2014 (HealthDay News) -- The U.S. outbreak of Enterovirus D68 has claimed its first confirmed victim, a 4- ...

114

Partial sequencing of the VP2 capsid gene for direct enterovirus genotyping in clinical specimens.  

PubMed

Typing of human enterovirus (EV) remains a major goal for diagnostic and epidemiological purposes. Whereas sequencing of the VP1 coding region is the reference standard for EV typing, a method relying on sequencing of the VP2 coding region has been proposed as an alternative; however, this has been validated only on cell culture supernatants. To avoid the selection of cultivable strains and to quicken the identification step, a new semi-nested PCR method targeting the VP2 region was developed by use of the CODEHOP strategy. After validation of the method on reference and clinical strains, a total of 352 clinical specimens found to be positive for EV RNA (138 with the GeneXpert EV kit and 214 with the Enterovirus R-gene kit) during a 3-year period (2010-2012) were analysed prospectively for VP2 genotyping. Overall, 204 (58%) specimens were typeable. A higher proportion of throat swab/stool specimens than of cerebrospinal fluid (CSF) specimens was found to be typeable (94 of 142 (66.2%) vs. 83 of 169 (49.1%), respectively, p<0.01 by the chi-square test). Moreover, the median Ct value obtained was lower for typeable specimens than for untypeable specimens (32.20 vs. 33.01, p<0.05, and 25.96 vs. 31.74, p<0.001, for the GeneXpert and R-gene tests, respectively, by the Mann-Whitney-Wilcoxon test). These results suggest that, in cases of EV meningitis, a peripheral specimen (i.e. throat swab or stool) that is susceptible to exhibiting a higher viral load should be used in preference to CSF for identifying the causative EV genotype by use of the VP2 typing method without cell culture isolation. PMID:24372815

Ibrahim, W; Boukhadra, N; Nasri-Zoghlami, D; Berthelot, P; Omar, S; Bourlet, T; Pozzetto, B; Pillet, S

2014-09-01

115

Human-Automation Interaction By Thomas B. Sheridan & Raja Parasuraman  

E-print Network

89 CHAPTER 2 Human-Automation Interaction By Thomas B. Sheridan & Raja Parasuraman Automation does not mean humans are replaced; quite the opposite. Increasingly, humans are asked to interact-free task for either the system designer or the human operator/automation supervisor, especially as computer

Parasuraman, Raja

116

A bird's eye view on human B cells  

Microsoft Academic Search

We show in this review that there is a continuum between the chicken B-cell system classified as the first GALT model described and the human B-cell system. We propose that humans have conserved for one B-cell subpopulation, the marginal zone B-cell subset in charge of T-independent responses, the strategies of diversification used by GALT species to generate their pre-immune repertoire.

Jean-Claude Weill; Sandra Weller; Claude-Agns Reynaud

2004-01-01

117

The ecology of enteroviruses in natural waters  

Microsoft Academic Search

More than 100 different enteric viruses are known to be excreted in human feces. More than 1 million viruses may be excreted per gram of feces, and concentrations as high as 500,000 infectious virus particles per liter have been detected in raw sewage. Certain enteric viruses can persist for long periods of time in the environment. Reported survival times range

Joseph L. Melnick; Charles P. Gerba; Gerald Berg

1980-01-01

118

Action of commonly used disinfectants against enteroviruses.  

PubMed

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

1983-06-01

119

A Novel Universal Neutralizing Monoclonal Antibody against Enterovirus 71 That Targets the Highly Conserved "Knob" Region of VP3 Protein  

PubMed Central

Hand, foot and mouth disease caused by enterovirus 71(EV71) leads to the majority of neurological complications and death in young children. While putative inactivated vaccines are only now undergoing clinical trials, no specific treatment options exist yet. Ideally, EV71 specific intravenous immunoglobulins could be developed for targeted treatment of severe cases. To date, only a single universally neutralizing monoclonal antibody against a conserved linear epitope of VP1 has been identified. Other enteroviruses have been shown to possess major conformational neutralizing epitopes on both the VP2 and VP3 capsid proteins. Hence, we attempted to isolate such neutralizing antibodies against conformational epitopes for their potential in the treatment of infection as well as differential diagnosis and vaccine optimization. Here we describe a universal neutralizing monoclonal antibody that recognizes a conserved conformational epitope of EV71 which was mapped using escape mutants. Eight escape mutants from different subgenogroups (A, B2, B4, C2, C4) were rescued; they harbored three essential mutations either at amino acid positions 59, 62 or 67 of the VP3 protein which are all situated in the knob region. The escape mutant phenotype could be mimicked by incorporating these mutations into reverse genetically engineered viruses showing that P59L, A62D, A62P and E67D abolish both monoclonal antibody binding and neutralization activity. This is the first conformational neutralization epitope mapped on VP3 for EV71. PMID:24875055

Meng, Tao; Chow, Vincent Tak Kwong; Kwang, Jimmy

2014-01-01

120

A novel universal neutralizing monoclonal antibody against enterovirus 71 that targets the highly conserved "knob" region of VP3 protein.  

PubMed

Hand, foot and mouth disease caused by enterovirus 71(EV71) leads to the majority of neurological complications and death in young children. While putative inactivated vaccines are only now undergoing clinical trials, no specific treatment options exist yet. Ideally, EV71 specific intravenous immunoglobulins could be developed for targeted treatment of severe cases. To date, only a single universally neutralizing monoclonal antibody against a conserved linear epitope of VP1 has been identified. Other enteroviruses have been shown to possess major conformational neutralizing epitopes on both the VP2 and VP3 capsid proteins. Hence, we attempted to isolate such neutralizing antibodies against conformational epitopes for their potential in the treatment of infection as well as differential diagnosis and vaccine optimization. Here we describe a universal neutralizing monoclonal antibody that recognizes a conserved conformational epitope of EV71 which was mapped using escape mutants. Eight escape mutants from different subgenogroups (A, B2, B4, C2, C4) were rescued; they harbored three essential mutations either at amino acid positions 59, 62 or 67 of the VP3 protein which are all situated in the "knob" region. The escape mutant phenotype could be mimicked by incorporating these mutations into reverse genetically engineered viruses showing that P59L, A62D, A62P and E67D abolish both monoclonal antibody binding and neutralization activity. This is the first conformational neutralization epitope mapped on VP3 for EV71. PMID:24875055

Kiener, Tanja K; Jia, Qiang; Meng, Tao; Chow, Vincent Tak Kwong; Kwang, Jimmy

2014-05-01

121

.. () B.B.A. (Human Resource Management) 895-171 (3)  

E-print Network

.. () B.B.A. (Human Resource Management) 135 1. 30 1.1 12 895-171 (3) Wisdom and Applications 460-211 (3) Operations Management 460-212 (3) Business Law 460-213 (3) Human Resource Resource Procurement 460-350 (3) Labor Relations 460-351 (3) Compensation Management 460-352 (3) Human

Laksanacharoen, Sathaporn

122

Bayesian modeling of human concept learning Joshua B. Tenenbaum  

E-print Network

Bayesian modeling of human concept learning Joshua B. Tenenbaum Department of Brain and Cognitive (Mitchell, 1997). This paper attempts to close the gap between human and machine concept learning dimension #12;+ + + + C - - - - + + + + + + + + + ++ + (a) (b) ( 0 c) Figure 1: (a) A rectangle concept

Tenenbaum, Josh

123

Bovine lactoferrin digested with human gastrointestinal enzymes inhibits replication of human echovirus 5 in cell culture.  

PubMed

Many infant formulas are enriched with lactoferrin (Lf) because of its claimed beneficial effects on health. Native bovine Lf (bLf) is known to inhibit in vitro replication of human enteroviruses, a group of pathogenic viruses that replicate in the gut as their primary infection site. On the basis of a model digestion and human gastrointestinal enzymes, we hypothesized that bLf could retain its antiviral properties against enterovirus in the gastrointestinal tract, either as an intact protein or through bioactive peptide fragments released by digestive enzymes. To test our hypothesis, bLf was digested with human gastric juice and duodenal juice in a 2-step in vitro digestion model. Two gastric pH levels and reduction conditions were used to simulate physiological conditions in adults and infants. The antiviral activity of native bLf and of the digested fractions was studied on echovirus 5 in vitro, using various assay conditions, addressing several mechanisms for replication inhibition. Both native and digested bLf fractions revealed a significant inhibitory effect, when added before or simultaneously with the virus onto the cells. Furthermore, a significant stronger sustained antiviral effect was observed when bLf was fully digested in the gastric phase with fast pH reduction to 2.5, compared with native bLf, suggesting the release of antiviral peptides from bLf during the human digestion process. In conclusion, this study demonstrates that bLf may have a role in the prevention of human gastrointestinal virus infection under physiological conditions and that food containing bLf may protect against infection in vivo. PMID:22901558

Furlund, Camilla B; Kristoffersen, Anja B; Devold, Tove G; Vegarud, Gerd E; Jonassen, Christine M

2012-07-01

124

Enterovirus 71 Can Directly Infect the Brainstem via Cranial Nerves and Infection Can Be Ameliorated by Passive Immunization.  

PubMed

Enterovirus 71 (EV71)-associated hand, foot, and mouth disease may be complicated by encephalomyelitis. We investigated EV71 brainstem infection and whether this infection could be ameliorated by passive immunization in a mouse model. Enterovirus 71 was injected into unilateral jaw/facial muscles of 2-week-old mice, and hyperimmune sera were given before or after infection. Harvested tissues were studied by light microscopy, immunohistochemistry, in situ hybridization, and viral titration. In unimmunized mice, viral antigen and RNA were detected within 24 hours after infection only in ipsilateral cranial nerves, motor trigeminal nucleus, reticular formation, and facial nucleus; viral titers were significantly higher in the brainstem than in the spinal cord samples. Mice given preinfection hyperimmune serum showed a marked reduction of ipsilateral viral antigen/RNA and viral titers in the brainstem in a dose-dependent manner. With optimum hyperimmune serum given after infection, brainstem infection was significantly reduced in a time-dependent manner. A delay in disease onset and a reduction of disease severity and mortality were also observed. Thus, EV71 can directly infect the brainstem, including the medulla, via cranial nerves, most likely by retrograde axonal transport. This may explain the sudden cardiorespiratory collapse in human patients with fatal encephalomyelitis. Moreover, our results suggest that passive immunization may still benefit EV71-infected patients who have neurologic complications. PMID:25289894

Tan, Soon Hao; Ong, Kien Chai; Wong, Kum Thong

2014-11-01

125

Detection of Enteroviruses and Rhinoviruses in Clinical Specimens by PCR and Liquid-Phase Hybridization  

Microsoft Academic Search

A sensitive method based on PCR followed by liquid-phase hybridization for detection of enterovirus and rhinovirus RNAs in clinical specimens and cell culture supernatants is described. RNA was extracted from stool samples, throat swabs, nasopharyngeal aspirates, cerebrospinal fluid, urine, and plasma with a com- mercial phenol-guanidinium-chloroform reagent and purified on a polysulfone membrane, on which the reversetranscriptasereactionwasalsodone.Twosetsofoligonucleotideprimersfromthe5*noncodingregion ofpicornaviruseswereselectedforDNAamplificationof153-bp(enterovirus)and120-bp(rhinovirus)regions. Double-stranded amplicons

PEKKA HALONEN; ELISABET ROCHA; JOHN HIERHOLZER; BRIAN HOLLOWAY; TIMO HYYPIA; PERTTI HURSKAINEN; ANDMARK PALLANSCH; Wallac Oy

1995-01-01

126

Identification and characterization of circulating human transitional B cells  

PubMed Central

Murine B-cell development begins in bone marrow and results in the generation of immature transitional B cells that transit to the spleen to complete their maturation. It remains unclear whether the same developmental pathway takes place in humans. Using markers characteristic of human bone marrow immature B cells, we have identified a population of circulating human B cells with a phenotype most similar to mouse transitional type I (T1) B cells, although these human counterparts express CD5. These cells die rapidly in culture, and B-cell activation factor member of the tumor necrosis factor (TNF) family (BAFF) does not effect their survival regardless of B-cell receptor (BCR) stimulation. In contrast, bone marrow stromal cells or interleukin-4 (IL-4) significantly enhanced their survival. In the presence of T-cell signals provided by IL-4 or CD40 ligation, BCR stimulation can induce progression into cell cycle. Interestingly, circulating B cells that phenotypically and functionally resemble murine T2 B cells are found in cord blood and adult peripheral blood, suggesting that B-cell maturation may not be restricted to the spleen. Notably, increased proportions of T1 B cells were found in blood of patients with systemic lupus erythematosus (SLE), although bone marrow production and selection appeared to be normal. PMID:15701725

Sims, Gary P.; Ettinger, Rachel; Shirota, Yuko; Yarboro, Cheryl H.; Illei, Gabor G.; Lipsky, Peter E.

2005-01-01

127

New insights in the regulation of human B cell differentiation  

PubMed Central

B lymphocytes provide the cellular basis of the humoral immune response. All stages of this process, from B cell activation to formation of germinal centers and differentiation into memory B cells or plasma cells, are influenced by extrinsic signals and controlled by transcriptional regulation. Compared to nave B cells, memory B cells display a distinct expression profile, which allows for their rapid secondary responses. Indisputably, many B cell malignancies result from aberrations in the circuitry controlling B cell function, particularly during the GC reaction. Here we review new insights into memory B cell subtypes, recent literature on transcription factors regulating human B cell differentiation, and further evidence for B cell lymphomagenesis emanating from errors during the GC cell reactions. PMID:19447676

Schmidlin, Heike; Diehl, Sean A.; Blom, Bianca

2009-01-01

128

Human Factors Analysis of Predator B Crash  

E-print Network

The 2006 crash of a Predator B in Arizona has prompted a great amount of scrutiny into Unmanned Arial System (UAS) operations. The direct cause of the Predator crash can be tied to an initial failure of the displays and a ...

Carrigan, Geoff

2008-06-10

129

Ultra Violet B Induced DNA Damage in Human Leucocyte Cells  

Microsoft Academic Search

The Single Cell Gel Electrophoresis (SCGE) Assay also known as Comet Assay is a rapid, simple and visual technique for assessing DNA damage in individual cells. Effect of UV-B was studied by irradiating human leucocytes. Irradiation of leucocyte increases the DNA instability, which can be assessed by Single Cell Gel Electrophoresis Assay. The blood samples were exposed to UV-B radiation

Abhay Singh Yadav; Manoj Kumar Sharma

2008-01-01

130

Streptozotocin is not toxic to the human fetal B cell  

Microsoft Academic Search

SummaryIt has been generally assumed that because streptozotocin is toxic to the adult B cell of most species, it should also damage B cells obtained at earlier stages of development. This paper examines whether this is true for human fetal pancreata obtained from the therapeutic termination of pregnancies during the first half of the second trimester. Experiments were carried out

B. E. Tuch; J. R. Turtle; C. J. Simeonovic

1989-01-01

131

Chlorogenic Acid Inhibits the Replication and Viability of Enterovirus 71 In Vitro  

PubMed Central

Enterovirus 71 (EV71) is an etiology for a number of diseases in humans. Traditional Chinese herbs have been reported to be effective for treating EV71 infection. However, there is no report about the antiviral effects of CHA against EV71. In this study, plaque reduction assay demonstrated that the inhibitory concentration 50% (IC50) of CHA on EV71 replication is 6.3 g/ml. When both CHA (20 g/ml) and EV71 were added, or added post-infection at different time points, CHA was able to effectively inhibit EV71 replication between 0 and 10 h. In addition, CHA inhibited EV71 2A transcription and translation in EV71-infected RD cells, but did not affect VP1, 3C, and 3D expression. Furthermore, CHA inhibited secretions of IL-6, TNF-?, IFN-? and MCP-1 in EV71-infected RD cells. Altogether, these results revealed that CHA may have antiviral properties for treating EV71 infection. PMID:24098754

Hou, Xueling; Peng, Hongjun; Zhang, Li; Shi, Mei; Ji, Yun; Wang, Yuyue

2013-01-01

132

Detection of enterovirus 71 using reverse transcription loop-mediated isothermal amplification (RT-LAMP).  

PubMed

Reverse transcription loop-mediated isothermal amplification (RT-LAMP), which is a visual assay for nucleic acids, is performed in a single step using one tube at 65 C for 1.5 h. In this study, RT-LAMP was established as a method for the detection of enterovirus 71 (EV71). The detection limit of the assay was approximately 10 copies, and no cross-reactivity was noted with Coxsackievirus A16, echovirus, human rotavirus (HRV) or norovirus. This assay, which offers greater sensitivity at a lower cost compared with the conventional reverse transcription polymerase chain reaction (RT-PCR), was validated using 252 clinical specimens that had been confirmed by laboratory diagnosis using RT-PCR. Both methods produced the same results with 52 positive samples. The RT-LAMP-based assay does not require specialised equipment, and therefore, it can be performed conveniently during an outbreak or under field conditions. In brief, the RT-LAMP-based assay provided a simple, rapid and efficient method for the detection of EV71 nucleic acid under field conditions. PMID:22155579

Wang, Xiang; Zhu, Jun-ping; Zhang, Qian; Xu, Zi-gang; Zhang, Fang; Zhao, Zhi-hui; Zheng, Wen-zhi; Zheng, Li-shu

2012-02-01

133

Rupintrivir is a promising candidate for treating severe cases of Enterovirus-71 infection  

PubMed Central

AIM: To evaluate the suitability of rupintrivir against Enterovirus 71 (EV71) induced severe clinical symptoms using computational methods. METHODS: The structure of EV71 3C protease was predicted by homology modeling. The binding free energies between rupintrivir and EV71 3C and human rhinovirus 3C protease were computed by molecular dynamics and molecular mechanics Poisson-Boltzmann/surface area and molecular mechanics generalized-born/surface area methods. EV71 3C fragments obtained from clinical samples collected during May to July 2008 in Shanghai were amplified by reverse-transcription and polymerase chain reaction and sequenced. RESULTS: We observed that rupintrivir had favorable binding affinity with EV71 3C protease (-10.76 kcal/mol). The variability of the 3C protein sequence in isolates of various outbreaks, including those obtained in our hospital from May to July 2008, were also analyzed to validate the conservation of the drug binding pocket. CONCLUSION: Rupintrivir, whose safety profiles had been proved, is an attractive candidate and can be quickly utilized for treating severe EV71 infection. PMID:20066739

Zhang, Xiao-Nan; Song, Zhi-Gang; Jiang, Ting; Shi, Bi-Sheng; Hu, Yun-Wen; Yuan, Zheng-Hong

2010-01-01

134

Antibody dependent enhancement infection of Enterovirus 71 in vitro and in vivo  

PubMed Central

Background Human enterovirus 71 (EV71) has emerged as a significant cause of acute encephalitis and deaths in young children. The clinical manifestations caused by EV71 varied from mild hand, foot and mouth disease to severe neurological complications and deaths, but its pathogenesis remains elusive. Antibody dependent enhancement (ADE) infection has been reported in various viruses and has been shown to contribute to disease severity. Results In this study, the presence of sub-neutralizing antibody was demonstrated to enhance EV71 infection in THP-1 cells and increase the mortality of EV71 infection in a suckling mouse model. Further, a secondary infection model was established to characterize the correlation between ADE and disease severity, and primary asymptomatic EV71 infection was shown to increase the mortality of the secondary EV71 infection in suckling mice. Conclusions Together, these in vitro and in vivo experiments strongly supported the hypothesis of ADE infection of EV71. The present findings indicate ADE might contribute to the pathogenesis of severe EV71 infection, and raise practical issues of vaccine development and antibody-based therapy. PMID:21385398

2011-01-01

135

Immunization of N terminus of enterovirus 71 VP4 elicits cross-protective antibody responses  

PubMed Central

Background Enterovirus 71 (EV71) is major cause of hand, foot and mouth disease. Large epidemics of EV71 infection have been recently reported in the Asian-Pacific region. Currently, no vaccine is available to prevent EV71 infection. Results The peptide (VP4N20) consisting of the first 20 amino acids at the N-terminal of VP4 of EV71 genotype C4 were fused to hepatitis B core (HBcAg) protein. Expression of fusion proteins in E. coli resulted in the formation of chimeric virus-like particles (VLPs). Mice immunized with the chimeric VLPs elicited anti-VP4N20 antibody response. In vitro microneutralization experiments showed that anti-chimeric VLPs sera were able to neutralize not only EV71 of genotype C4 but also EV71 of genotype A. Neonatal mice model confirmed the neutralizing ability of anti-chimeric VLPs sera. Eiptope mapping led to the identification of a core sequence responsible for antibody recognition within the peptide. Conclusions Immunization of chimeric VLPs is able to elicit antibodies displaying a broad neutralizing activity against different genotypes of EV71 in vitro. The core sequence of EV71-VP4 is highly conserved across EV71 genotypes. The chimeric VLPs have a great potential to be a novel vaccine candidate with a broad cross-protection against different EV71 genotypes. PMID:24320792

2013-01-01

136

3C Protease of Enterovirus 68: Structure-Based Design of Michael Acceptor Inhibitors and Their Broad-Spectrum Antiviral Effects  

E-print Network

3C Protease of Enterovirus 68: Structure-Based Design of Michael Acceptor Inhibitors have determined the cleavage specificity and the crystal structure of the 3C protease of enterovirus 68 on the one hand and classical enteroviruses on the other prompted us to use the crystal structure

Lübeck, Universität zu

137

Detection of Astroviruses, Enteroviruses, and Adenovirus Types 40 and 41 in Surface Waters Collected and Evaluated by the Information Collection Rule and an Integrated Cell Culture-Nested PCR Procedure  

Microsoft Academic Search

We evaluated the use of an integrated cell culture-reverse transcription-PCR (ICC-RT-PCR) procedure coupled with nested PCR to detect human astroviruses, enteroviruses, and adenovirus types 40 and 41 in surface water samples that were collected and evaluated by using the Information Collection Rule (ICR) method. The results obtained with the ICC-RT-PCR-nested PCR method were compared to the results obtained with the

CHRISTOPHER D. CHAPRON; NICOLA A. BALLESTER; JUSTIN H. FONTAINE; CHRISTINE N. FRADES; AARON B. MARGOLIN

2000-01-01

138

Age effects on B cells and humoral immunity in humans  

PubMed Central

Both humoral and cellular immune responses are impaired in aged individuals, leading to decreased vaccine responses. Although T cell defects occur, defects in B cells play a significant role in age-related humoral immune changes. The ability to undergo class switch recombination (CSR), the enzyme for CSR, AID (activation-induced cytidine deaminase) and the transcription factor E47 are all decreased in aged stimulated B cells. We here present an overview of age-related changes in human B cell markers and functions, and also discuss some controversies in the field of B cell aging. PMID:20728581

Frasca, Daniela; Diaz, Alain; Romero, Maria; Landin, Ana Marie; Blomberg, Bonnie B

2010-01-01

139

Age effects on B cells and humoral immunity in humans.  

PubMed

Both humoral and cellular immune responses are impaired in aged individuals, leading to decreased vaccine responses. Although T cell defects occur, defects in B cells play a significant role in age-related humoral immune changes. The ability to undergo class switch recombination (CSR), the enzyme for CSR, AID (activation-induced cytidine deaminase) and the transcription factor E47 are all decreased in aged stimulated B cells. We here present an overview of age-related changes in human B cell markers and functions, and also discuss some controversies in the field of B cell aging. PMID:20728581

Frasca, Daniela; Diaz, Alain; Romero, Maria; Landin, Ana Marie; Blomberg, Bonnie B

2011-07-01

140

Molecular analysis of enterovirus in Cameroon by partial 5'UTR-VP4 gene sequencing reveals a high genetic diversity and frequency of infections.  

PubMed

Enteroviruses (EVs) often infect humans, presenting as endemic or epidemic infections. In this study, the diversity of infecting EVs was studied among 146 children and 137 adults in a small community in Cameroon. The participants provided 2,458 fecal samples during 1-year monthly collection; 10 or more samples were obtained from 55%. Partial 5'UTR-VP4 region could be sequenced in 393/547 PCR positive samples obtained from 119 children and 85 adults. EV-RNA was detected in at least one sample from 235 participants (83%) during the study period. A total of 121 different strains were identified, 66 infected only children, 29 only adults, and 26 infected both children and adults. There were children with up to five episodes with different strains, and adults with up to four such episodes. Infants aged <5 years were significantly more often EV infected compared to older participants. Infections with species EV-C constituted two third of all cases, and overall EV infections were more common during the rainy season. Species EV-B more often infected children than adults. Most strains were detected only for certain months of the year; however five strains were observed during the time spans of 5-10 months. Two strains were excreted up to eight months in three children and one adult. In 11 of the 128 families with paired samples the child and the adult were infected simultaneously by the same strain, indicating common source of infection. The study revealed a surprising complexity of EV ecosystem in a single community. J. Med. Virol. 86:2092-2101, 2014. 2014 Wiley Periodicals, Inc. PMID:24634190

Ayukekbong, James Ayukepi; Fobisong, Cajetan; Lindh, Magnus; Nkuo-Akenji, Theresia; Bergstrm, Tomas; Norder, Helene

2014-12-01

141

Comparative Study of the Cytokine/Chemokine Response in Children with Differing Disease Severity in Enterovirus 71-Induced Hand, Foot, and Mouth Disease  

PubMed Central

Background Enterovirus 71 (EV71) infection can lead to a rapidly progressing, life-threatening, and severe neurological disease in young children, including the development of human hand, foot, and mouth disease (HFMD). This study aims to further characterize the specific immunological features in EV71mediated HFMD patients presenting with differing degrees of disease severity. Methodology Comprehensive cytokine and chemokine expression were broadly evaluated by cytokine antibody array in EV71infected patients hospitalized for HFMD compared to Coxsackievirus A16-infected patients and age-matched healthy controls. More detailed analysis using Luminex-based cytokine bead array was performed in EV71infected patients stratified into diverse clinic outcomes. Additionally, immune cell frequencies in peripheral blood and EV71specific antibodies in plasma were also examined. Principal Findings Expression of several cytokines and chemokines were significantly increased in plasma from EV71infected patients compared to healthy controls, which further indicated that: (1) GM-CSF, MIP-1?, IL-2, IL-33, and IL-23 secretion was elevated in patients who rapidly developed disease and presented with uncomplicated neurological damage; (2) G-CSF and MCP-1 were distinguishably secreted in EV71 infected very severe patients presenting with acute respiratory failure; (3) IP-10, MCP-1, IL-6, IL-8, and G-CSF levels were much higher in cerebrospinal fluid than in plasma from patients with neurological damage; (4) FACS analysis revealed that the frequency of CD19+HLADR+ mature B cells dynamically changed over time during the course of hospitalization and was accompanied by dramatically increased EV71specific antibodies. Our data provide a panoramic view of specific immune mediator and cellular immune responses of HFMD and may provide useful immunological profiles for monitoring the progress of EV71induced fatal neurological symptoms with acute respiratory failure. PMID:23840697

Wang, Linghang; Yang, Fan; Hu, Yongfeng; Ren, Xianwen; Li, Guojun; Yu, Yang; Sun, Shaoxia; Li, Yufen; Chen, Xinchun; Li, Xingwang; Jin, Qi

2013-01-01

142

Monoamine oxidase B activity is increased in human gliomas.  

PubMed

Glial tumours are the most common type of brain neoplasm in humans. Tumour classification and grading represent key factors for patient management. However, current grading schemes are still limited by subjective histological criteria. In this context, gliosis has been linked to increases in monoamine oxidase B (MAO-B) activity. Thus, in the present study, MAO-B activity in membranes of glial tumours (n=20), meningiomas (n=12) and non-pathological human brains (n=15) was quantified by [14C]PEA oxidation. MAO-B activity was significantly greater in glioblastoma multiformes than in postmortem control brains (p<0.01) or meningiomas (p<0.001). There were no significant differences in MAO-B activity between glioblastoma multiformes (n=11) and low-grade astrocytomas (n=3) or anaplastic astrocytomas (n=6). In conclusion, the present results demonstrate a significant and selective increase in MAO-B activity in human gliomas when compared with meningiomas or non-tumoural tissue. These results suggest that the quantification of MAO-B activity may be a useful diagnostic tool for differentiating glial tumours from other types of brain tumours or surrounding normal brain tissue. PMID:17624626

Gabilondo, Ane M; Hostalot, Cristina; Garibi, Jess M; Meana, J Javier; Callado, Luis F

2008-01-01

143

Cell Surface Vimentin Is an Attachment Receptor for Enterovirus 71  

PubMed Central

ABSTRACT Enterovirus 71 (EV71) is a highly transmissible pathogenic agent that causes severe central nervous system diseases in infected infants and young children. Here, we reported that EV71 VP1 protein could bind to vimentin intermediate filaments expressed on the host cell surface. Soluble vimentin or an antibody against vimentin could inhibit the binding of EV71 to host cells. Accompanied with the reduction of vimentin expression on the cell surface, the binding of EV71 to cells was remarkably decreased. Further evidence showed that the N terminus of vimentin is responsible for the interaction between EV71 and vimentin. These results indicated that vimentin on the host cell surface may serve as an attachment site that mediated the initial binding and subsequently increased the infectivity of EV71. IMPORTANCE This study delivers important findings on the roles of vimentin filaments in relation to EV71 infection and provides information that not only improves our understanding of EV71 pathogenesis but also presents us with potentially new strategies for the treatment of diseases caused by EV71 infections. PMID:24623428

Du, Ning; Cong, Haolong; Tian, Hongchao; Zhang, Hua; Zhang, Wenliang; Song, Lei

2014-01-01

144

Bcl-B Expression in Human Epithelial and Nonepithelial Malignancies  

PubMed Central

Purpose Apoptosis plays an important role in neoplastic processes. Bcl-B is an antiapoptotic Bcl-2 family member, which is known to change its phenotype upon binding to Nur77/TR3. The expression pattern of this protein in human malignancies has not been reported. Experimental Design We investigated Bcl-B expression in normal human tissues and several types of human epithelial and nonepithelial malignancy by immunohistochemistry, correlating results with tumor stage, histologic grade, and patient survival. Results Bcl-B protein was strongly expressed in all normal plasma cells but found in only18%of multiple myelomas (n = 133). Bcl-B immunostaining was also present in normal germinal center centroblasts and centrocytes and in approximately half of diffuse large B-cell lymphoma (n =48) specimens, whereas follicular lymphomas (n = 57) did not contain Bcl-B. In breast (n = 119), prostate (n = 66), gastric (n = 180), and colorectal (n = 106) adenocarcinomas, as well as in non small cell lung cancers (n = 82), tumor-specific overexpression of Bcl-B was observed. Bcl-B expression was associated with variables of poor prognosis, such as high tumor grade in breast cancer (P = 0.009), microsatellite stability (P = 0.0002), and left-sided anatomic location (P = 0.02) of colorectal cancers, as well as with greater incidence of death from prostate cancer (P = 0.005) and shorter survival of patients with small cell lung cancer (P = 0.009). Conversely, although overexpressed in many gastric cancers, Bcl-B tended to correlate with better outcome (P = 0.01) and more differentiated tumor histology (P < 0.0001). Conclusions Tumor-specific alterations in Bcl-B expressionmay define subsets of nonepithelial and epithelial neoplasms with distinct clinical behaviors. PMID:18483366

Krajewska, Maryla; Kitada, Shinichi; Winter, Jane N.; Variakojis, Daina; Lichtenstein, Alan; Zhai, Dayong; Cuddy, Michael; Huang, Xianshu; Luciano, Frederic; Baker, Cheryl H.; Kim, Hoguen; Shin, Eunah; Kennedy, Susan; Olson, Allen H.; Badzio, Andrzej; Jassem, Jacek; Meinhold-Heerlein, Ivo; Duffy, Michael J.; Schimmer, Aaron D.; Tsao, Ming; Brown, Ewan; Sawyers, Anne; Andreeff, Michael; Mercola, Dan; Krajewski, Stan; Reed, John C.

2014-01-01

145

CD19 and CD32b differentially regulate human B cell responsiveness.  

PubMed

B cell activation is regulated by a variety of signals. CD19 positively regulates B cell activation, augmenting signals delivered through the BCR complex. In contrast, CD32b contains an ITIM and negatively regulates BCR signaling. Importantly, there are drugs currently in clinical trials and preclinical development that cross-link CD32b to molecules within the BCR complex. We wanted to address how single engagement versus cotargeting these molecules affects human B cell function. When B cells from healthy individuals were activated by signals that mimic a T cell response (IL-21 costimulation), ligation of CD32b, but not CD19, inhibited B cell expansion and plasma cell (PC) differentiation. In contrast, when B cells were activated through TLR, anti-CD19, but not anti-CD32b, blunted the response. However, when both CD19 and CD32b were coengaged by a bispecific anti-CD19CD32b Ab, both types of stimuli were potently inhibited. Cross-linking CD19 with CD32b also inhibited Ab-independent functions of B cells, such as HLA upregulation, cytokine production, and the ability of B cells to prime CD4(+) T cells. Finally, although cross-linking CD19 and CD32b inhibited PC differentiation of primary B cells, it did not alter Ig production from pre-established PCs. These data elucidate the mechanism by which a complex set of signals determines the fate of B cell responsiveness. Although signals through CD19 influence TLR-driven activation, CD32b impacts the magnitude of the response following IL-21 costimulation. Therefore, simultaneous targeting of multiple surface molecules may be a necessary approach to comprehensively modulate B cell activation in vivo. PMID:24442430

Karnell, Jodi L; Dimasi, Nazzareno; Karnell, Fredrick G; Fleming, Ryan; Kuta, Ellen; Wilson, Mildred; Wu, Herren; Gao, Changshou; Herbst, Ronald; Ettinger, Rachel

2014-02-15

146

Exploiting Human Memory B Cell Heterogeneity for Improved Vaccine Efficacy  

PubMed Central

The major goal in vaccination is establishment of long-term, prophylactic humoral memory to a pathogen. Two major components to long-lived humoral memory are plasma cells for the production of specific immunoglobulin and memory B cells that survey for their specific antigen in the periphery for later affinity maturation, proliferation, and differentiation. The study of human B cell memory has been aided by the discovery of a general marker for B cell memory, expression of CD27; however, new data suggests the existence of CD27? memory B cells as well. These recently described non-canonical memory populations have increasingly pointed to the heterogeneity of the memory compartment. The novel B memory subsets in humans appear to have unique origins, localization, and functions compared to what was considered to be a classical memory B cell. In this article, we review the known B cell memory subsets, the establishment of B cell memory in vaccination and infection, and how understanding these newly described subsets can inform vaccine design and disease treatment. PMID:22566866

Pauli, Noel T.; Henry Dunand, Carole J.; Wilson, Patrick C.

2011-01-01

147

Biochemistry of B12-cofactors in human metabolism.  

PubMed

Vitamin B12, the "antipernicious anaemia factor", is a crystallisable cobalt-complex, which belongs to a group of unique "complete" corrinoids, named cobalamins (Cbl). In humans, instead of the "vitamin", two organometallic B12-forms are coenzymes in two metabolically important enzymes: Methyl-cobalamin, the cofactor of methionine synthase, and coenzyme B12 (adenosyl-cobalamin), the cofactor of methylmalonyl-CoA mutase. The cytoplasmatic methionine synthase catalyzes the transfer of a methyl group from N-methyl-tetrahydrofolate to homocysteine to yield methionine and to liberate tetrahydrofolate. In the mitochondrial methylmalonyl-CoA mutase a radical process transforms methylmalonyl-CoA (a remains e.g. from uneven numbered fatty acids) into succinyl-CoA, for further metabolic use. In addition, in the human mitochondria an adenosyl-transferase incorporates the organometallic group of coenzyme B12. In all these enzymes, the bound B12-derivatives engage (or are formed) in exceptional organometallic enzymatic reactions. This chapter recapitulates the physiological chemistry of vitamin B12, relevant in the context of the metabolic transformation of B12-derivatives into the relevant coenzyme forms and their use in B12-dependent enzymes. PMID:22116707

Krutler, Bernhard

2012-01-01

148

Disulfide Bond Configuration of Human Cytomegalovirus Glycoprotein B  

PubMed Central

Glycoprotein B (gB) is the most highly conserved of the envelope glycoproteins of human herpesviruses. The gB protein of human cytomegalovirus (CMV) serves multiple roles in the life cycle of the virus. To investigate structural properties of gB that give rise to its function, we sought to determine the disulfide bond arrangement of gB. To this end, a recombinant form of gB (gB-S) comprising the entire ectodomain of the glycoprotein (amino acids 1 to 750) was constructed and expressed in insect cells. Proteolytic fragmentation and mass spectrometry were performed using purified gB-S, and the five disulfide bonds that link 10 of the 11 highly conserved cysteine residues of gB were mapped. These bonds are C94-C550, C111-C506, C246-C250, C344-C391, and C573-C610. This configuration closely parallels the disulfide bond configuration of herpes simplex type 2 (HSV-2) gB (N. Norais, D. Tang, S. Kaur, S. H. Chamberlain, F. R. Masiarz, R. L. Burke, and F. Markus, J. Virol. 70:7379-7387, 1996). However, despite the high degree of conservation of cysteine residues between CMV gB and HSV-2 gB, the disulfide bond arrangements of the two homologs are not identical. We detected a disulfide bond between the conserved cysteine residue 246 and the nonconserved cysteine residue 250 of CMV gB. We hypothesize that this disulfide bond stabilizes a tight loop in the amino-terminal fragment of CMV gB that does not exist in HSV-2 gB. We predicted that the cysteine residue not found in a disulfide bond of CMV gB, cysteine residue 185, would play a role in dimerization, but a cysteine substitution mutant in cysteine residue 185 showed no apparent defect in the ability to form dimers. These results indicate that gB oligomerization involves additional interactions other than a single disulfide bond. This work represents the second reported disulfide bond structure for a herpesvirus gB homolog, and the discovery that the two structures are not identical underscores the importance of empirically determining structures even for highly conserved proteins. PMID:12021340

Lopper, Matthew; Compton, Teresa

2002-01-01

149

Disulfide bond configuration of human cytomegalovirus glycoprotein B.  

PubMed

Glycoprotein B (gB) is the most highly conserved of the envelope glycoproteins of human herpesviruses. The gB protein of human cytomegalovirus (CMV) serves multiple roles in the life cycle of the virus. To investigate structural properties of gB that give rise to its function, we sought to determine the disulfide bond arrangement of gB. To this end, a recombinant form of gB (gB-S) comprising the entire ectodomain of the glycoprotein (amino acids 1 to 750) was constructed and expressed in insect cells. Proteolytic fragmentation and mass spectrometry were performed using purified gB-S, and the five disulfide bonds that link 10 of the 11 highly conserved cysteine residues of gB were mapped. These bonds are C94-C550, C111-C506, C246-C250, C344-C391, and C573-C610. This configuration closely parallels the disulfide bond configuration of herpes simplex type 2 (HSV-2) gB (N. Norais, D. Tang, S. Kaur, S. H. Chamberlain, F. R. Masiarz, R. L. Burke, and F. Markus, J. Virol. 70:7379-7387, 1996). However, despite the high degree of conservation of cysteine residues between CMV gB and HSV-2 gB, the disulfide bond arrangements of the two homologs are not identical. We detected a disulfide bond between the conserved cysteine residue 246 and the nonconserved cysteine residue 250 of CMV gB. We hypothesize that this disulfide bond stabilizes a tight loop in the amino-terminal fragment of CMV gB that does not exist in HSV-2 gB. We predicted that the cysteine residue not found in a disulfide bond of CMV gB, cysteine residue 185, would play a role in dimerization, but a cysteine substitution mutant in cysteine residue 185 showed no apparent defect in the ability to form dimers. These results indicate that gB oligomerization involves additional interactions other than a single disulfide bond. This work represents the second reported disulfide bond structure for a herpesvirus gB homolog, and the discovery that the two structures are not identical underscores the importance of empirically determining structures even for highly conserved proteins. PMID:12021340

Lopper, Matthew; Compton, Teresa

2002-06-01

150

Acute human parvovirus b19 infection: cytologic diagnosis.  

PubMed

Human parvovirus B19 is highly tropic to human bone marrow and replicates only in erythroid progenitor cells. It is causative agent of transient aplastic crisis in patients with chronic haemolytic anemia. In immunocompromised patients persistent parvovirus B19 infection may develop and it manifests as pure red cell aplasia and chronic anaemia. Bone marrow is characterised morphologically by giant pronormoblast stage with little or no further maturation. We encountered a case of 6year old HIV positive male child presented with pure red cell aplasia due to parvovirus B19 infection. Bone marrow aspiration cytology revealed giant pronormoblast with prominent intranuclear inclusions led to suspicion of parvovirus B19 infection which was confirmed by DNA PCR. This case is presented to report classical morphological features of parvovirus B19 infection rarely seen on bone marrow examination should warrant the suspicion of human parvovirus B19 infection in the setting of HIV positive patient with repeated transfusions and confirmation should be done by PCR. PMID:25332559

Sharada Raju, Rane; Nalini Vinayak, Kadgi; Madhusudan Bapat, Vishnuprasad; Preeti Balkisanji, Agrawal; Shaila Chandrakant, Puranik

2014-09-01

151

Non-polio enteroviruses and their association with acute diarrhea in children in India.  

PubMed

A causative agent in approximately 40% of diarrheal cases still remains unidentified. Though many enteroviruses (EVs) are transmitted through fecal-oral route and replicate in the intestinal cells, their association with acute diarrhea has not so far been recognized due to lack of detailed epidemiological investigations. This long-term, detailed molecular epidemiological study aims to conclusively determine the association of non-polio enteroviruses (NPEVs) with acute diarrhea in comparison with rotavirus (RV) in children. Diarrheal stool specimens from 2161 children aged 0-2 years and 169 children between 2 and 9 years, and 1800 normal stool samples from age-matched healthy children between 0 and 9 years were examined during 2008-2012 for enterovirus (oral polio vaccine strains (OPVs) and NPEVs). Enterovirus serotypes were identified by complete VP1 gene sequence analysis. Enterovirus and rotavirus were detected in 19.01% (380/2330) and 13.82% (322/2330) diarrheal stools. During the study period, annual prevalence of EV- and RV-associated diarrhea ranged between 8% and 22%, but with contrasting seasonal prevalence with RV predominating during winter months and NPEV prevailing in other seasons. NPEVs are associated with epidemics-like outbreaks during which they are detected in up to 50% of diarrheic children, and in non-epidemic seasons in 0-10% of the patients. After subtraction of OPV-positive diarrheal cases (1.81%), while NPEVs are associated with about 17% of acute diarrhea, about 6% of healthy children showed asymptomatic NPEV excretion. Of 37 NPEV serotypes detected in diarrheal children, seven echovirus types 1, 7, 11, 13, 14, 30 and 33 are frequently observed, with E11 being more prevalent followed by E30. In conclusion, NPEVs are significantly associated with acute diarrhea, and NPEVs and rotavirus exhibit contrasting seasonal predominance. This study signifies the need for a new direction of research on enteroviruses involving systematic analysis of their contribution to diarrheal burden. PMID:23602838

Rao, Durga C; Ananda Babu, M; Raghavendra, A; Dhananjaya, D; Kumar, Sudheendra; Maiya, P P

2013-07-01

152

Human kidney anion exchanger 1 interacts with kinesin family member 3B (KIF3B)  

SciTech Connect

Highlights: {yields} Impaired trafficking of kAE1 causes distal renal tubular acidosis (dRTA). {yields} The interaction between kAE1 and kinesin family member 3B (KIF3B) is reported. {yields} The co-localization between kAE and KIF3B was detected in human kidney tissues. {yields} A marked reduction of kAE1 on the cell membrane was observed when KIF3B was knockdown. {yields} KFI3B plays an important role in trafficking of kAE1 to the plasma membrane. -- Abstract: Impaired trafficking of human kidney anion exchanger 1 (kAE1) to the basolateral membrane of {alpha}-intercalated cells of the kidney collecting duct leads to the defect of the Cl{sup -}/HCO{sub 3}{sup -} exchange and the failure of proton (H{sup +}) secretion at the apical membrane of these cells, causing distal renal tubular acidosis (dRTA). In the sorting process, kAE1 interacts with AP-1 mu1A, a subunit of AP-1A adaptor complex. However, it is not known whether kAE1 interacts with motor proteins in its trafficking process to the plasma membrane or not. We report here that kAE1 interacts with kinesin family member 3B (KIF3B) in kidney cells and a dileucine motif at the carboxyl terminus of kAE1 contributes to this interaction. We have also demonstrated that kAE1 co-localizes with KIF3B in human kidney tissues and the suppression of endogenous KIF3B in HEK293T cells by small interfering RNA (siRNA) decreases membrane localization of kAE1 but increases its intracellular accumulation. All results suggest that KIF3B is involved in the trafficking of kAE1 to the plasma membrane of human kidney {alpha}-intercalated cells.

Duangtum, Natapol [Medical Molecular Biology Unit, Office for Research and Development Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700 (Thailand) [Medical Molecular Biology Unit, Office for Research and Development Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700 (Thailand); Department of Anatomy, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700 (Thailand); Junking, Mutita; Sawasdee, Nunghathai [Medical Molecular Biology Unit, Office for Research and Development Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700 (Thailand)] [Medical Molecular Biology Unit, Office for Research and Development Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700 (Thailand); Cheunsuchon, Boonyarit [Department of Pathology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700 (Thailand)] [Department of Pathology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700 (Thailand); Limjindaporn, Thawornchai, E-mail: limjindaporn@yahoo.com [Department of Anatomy, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700 (Thailand)] [Department of Anatomy, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700 (Thailand); Yenchitsomanus, Pa-thai, E-mail: grpye@mahidol.ac.th [Medical Molecular Biology Unit, Office for Research and Development Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700 (Thailand)] [Medical Molecular Biology Unit, Office for Research and Development Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700 (Thailand)

2011-09-16

153

Cytotoxicity of (-)-vitisin B in human leukemia cells.  

PubMed

Vitis thunbergii var. taiwaniana (VTT) is an indigenous Taiwanese wild grape and is used as a folk medicine in Taiwan. VTT is rich in polyphenols, especially quercetin and resveratrol derivatives, which were demonstrated to exhibit inhibitory activities against carcinogenesis and prevent some neurodegenerative diseases. (-)-Vitisin B is one of the resveratrol tetramers extracted from VTT. In this study, we investigated the mechanisms of (-)-vitisin B on the induction of apoptosis in human HL-60 promyelocytic leukemia cells. First, (-)-vitisin B significantly inhibited cell proliferation through inducing cell apoptosis. This effect appeared to occur in a time- and dose-dependent manner. Cell-cycle distribution was also examined, and we found that (-)-vitisin B significantly induced a sub-G1 population in a dose-dependent manner. In addition, (-)-vitisin B exhibited stronger inhibitory effects on cell proliferation than resveratrol. Second, (-)-vitisin B dose dependently induced apoptosis-related protein expressions, such as the cleavage form of caspase-3, caspase-8, caspase-9, poly(ADP ribose) polymerase, and the proapoptotic Bax protein. Third, (-)-vitisin B treatment also resulted in increases in c-Jun N-terminal kinase (JNK) phosphorylation and Fas ligand (FasL) expression. Moreover, the (-)-vitisin B-induced FasL expression and caspase-3 activation could be reversed by a JNK inhibitor. These results suggest that (-)-vitisin B-induced apoptosis of leukemia cells might be mediated through activation of JNK and Fas death-signal transduction. PMID:23030068

Wu, Shing-Sheng; Chen, Lih-Geeng; Lin, Ren-Jye; Lin, Shyr-Yi; Lo, Yueh-E; Liang, Yu-Chih

2013-07-01

154

Molecular Analysis of Virulent Determinants of Enterovirus 71  

PubMed Central

Enterovirus 71 (EV71) is the most important causative agent of hand, foot and mouth disease (HFMD) in children. In most cases, it is a self-limiting illness. However some EV71 infectious cases can develop severe clinical outcomes, such as encephalitis, meningitis, poliomyelitis like paralysis, and even death. To identify the determinants of virulence, the deduced amino acid sequence of polyprotein and nucleotide sequence of 5?-NTR and 3?-NTR in 25 SC-EV71 strains (strains from severe cases) and 31 MC-EV71 strains (strains from mild cases) were analyzed. Results showed four amino acids on two positions (GlyP710/GlnP710/ArgP710 and GluP729) on the DE and EF loop of VP1, one (LysP930) on the surface of protease 2A and four nucleotides on three positions (GP272, UP488 and AP700/UP700) in the 5'-NTR region are associated with EV71 virulent phenotype. Predicted secondary structure of RNA using the consensus sequence of 5'-NTR by RNAStructure showed the mutation of nucleotide at position 488 in strain BJ08-Z004-3 (position 491 in prototype strain BrCr) can result in the discrepancy of an additional pair of nucleotides and thus change the stability of the second structure of IRES. Fragment base content analysis showed that in the region 696 to 714 bp at the 5'-NTR, where the AP700/UP700 was located, the nucleotide constitution ratios differed significantly between SC-EV71 and MC-EV71 strains. In conclusion, comparative genomic analysis showed that virulence of EV71 strains are mainly determined by the amino acids on two positions of VP1, one position of protease 2A and the nucleotides on three positions in 5'-NTR. PMID:22039449

Chen, Lijuan; Zhang, Herun; Wang, Yumei

2011-01-01

155

Enterovirus 71 infection: An experience in Korea, 2009.  

PubMed

Enterovirus 71 (EV71) has been recognized as a frequent cause of epidemics of hand-foot-and-mouth disease (HFMD) associated with severe neurological symptoms. In the spring of 2009, HFMD was epidemic in Korea. Severe cases with complication, including death, have been reported and it has become a public health issue. Most symptomatic EV71 infections commonly result in HFMD or herpangina. These clinical manifestations can be associated with neurologic syndromes frequently. Neurologic syndromes observed in EV71 include meningitis, meningoencephalomyelitis, poliomyelitis-like paralytic disease, Guillain-Barr syndrome, transverse myelitis, cerebellar ataxia, opsoclonus-myoclonus syndrome, benign intracranial hypertension, and brainstem encephalitis. Examinations for EV 71 were performed from the stools, respiratory secretion or CSF of the children by realtime PCR. Gene analysis showed that most of them were caused by EV71 subgenotype C4a which was prevalent in China, 2008. Public health measures including personal and environmental hygiene, must to target daycare centers, kindergartens, and schools where highly susceptible children congregate. To prevent the spread of infection, preschools where transmission persists for more than 2 incubation periods, have been recommended for closure, and trigger criteria for voluntary closure was instituted. During closure, operators are to thoroughly clean the centers before they are allowed to reopen. In addition, parents are advised to ensure that their children adopt a high standard of personal hygiene and to keep the infected child at home until full recovery. Because the outbreaks occur in a cyclical pattern, surveillance system to predict next outbreaks and adequate public health measures to control need to be planned for future. Control of EV71 epidemics through surveillance and public health intervention needs to be maintained in Korea. Future research should focus on understanding of EV71 virulence, identification of the receptor(s) for EV71, development of antiviral agents and development of vaccine. PMID:21189926

Kim, Kyung Hyo

2010-05-01

156

Implication of Human UGT2B7, 2B15, and 2B17 in 19-Norandrosterone Metabolism  

PubMed Central

Nandrolone (19-nortestosterone) is an anabolic androgenic steroid commonly abused for doping purposes. Nandrolone is mainly metabolized in the liver into 19-norandrosterone prior to glucuronidation and excretion through urine over an extended period of time. Several UGTs (i.e., UGT2B7, UGT2B15, and UGT2B17) are thought to be the major enzymes responsible for conjugation of androgens in human. An in vitro study using recombinant enzymes expressed in insect cells showed that UGT1A4 and UGT2B7 are the two main enzymes responsible of 19-norandrosterone glucuronidation. However, the identity of the enzyme involved in nandrolone metabolism in vivo together with their relative contribution and regulation remain unknown. Inhibition assays using human liver microsomes (HLM) incubated with 19-norandrosterone and selective inhibitors confirmed that UGT2B7 and UGT2B15 are involved in 19-norandrosterone glucuronidation, since the presence of the specific UGT2B7 and UGT2B15 inhibitors gemfibrozil and valproic acid inhibited the 19-norandrosterone glucuronidation by 35 and 45%, respectively. HLM were genotyped for UGT2B15 D85Y, UGT2B7 H268Y, and the UGT2B17 deletion polymorphism. The glucuronidation activity on 19-norandrosterone was significantly higher in UGT2B15 DD than in the other UGT2B15 genotypes (p?human liver cancer HepG2 cells were exposed to androgens to determine if the transcriptional activity of the genes of interest was affected. Only UGT2B7 mRNA expression was significantly increased (1.8-folds) after incubation with nandrolone decanoate. These results show that the UGT2B7 and UGT2B15 are involved in 19-norandrosterone glucuronidation and that the UGT2B15 polymorphism (D85Y) is the only UGT genetic variation that influences the glucuronidation activity. This could partly explain the inter-individual variation in 19-norandrosterone excretion. PMID:23805127

Strahm, Emmanuel; Sjoberg, Ulf; Garle, Mats; Rane, Anders; Ekstrom, Lena

2013-01-01

157

Structures of the Procapsid and Mature Virion of Enterovirus 71 Strain 1095  

PubMed Central

Enterovirus 71 (EV71) is an important emerging human pathogen with a global distribution and presents a disease pattern resembling poliomyelitis with seasonal epidemics that include cases of severe neurological complications, such as acute flaccid paralysis. EV71 is a member of the Picornaviridae family, which consists of icosahedral, nonenveloped, single-stranded RNA viruses. Here we report structures derived from X-ray crystallography and cryoelectron microscopy (cryo-EM) for the 1095 strain of EV71, including a putative precursor in virus assembly, the procapsid, and the mature virus capsid. The cryo-EM map of the procapsid provides new structural information on portions of the capsid proteins VP0 and VP1 that are disordered in the higher-resolution crystal structures. Our structures solved from virus particles in solution are largely in agreement with those from prior X-ray crystallographic studies; however, we observe small but significant structural differences for the 1095 procapsid compared to a structure solved in a previous study (X. Wang, W. Peng, J. Ren, Z. Hu, J. Xu, Z. Lou, X. Li, W. Yin, X. Shen, C. Porta, T. S. Walter, G. Evans, D. Axford, R. Owen, D. J. Rowlands, J. Wang, D. I. Stuart, E. E. Fry, and Z. Rao, Nat. Struct. Mol. Biol. 19:424429, 2012) for a different strain of EV71. For both EV71 strains, the procapsid is significantly larger in diameter than the mature capsid, unlike in any other picornavirus. Nonetheless, our results demonstrate that picornavirus capsid expansion is possible without RNA encapsidation and that picornavirus assembly may involve an inward radial collapse of the procapsid to yield the native virion. PMID:23637404

Cifuente, Javier O.; Lee, Hyunwook; Yoder, Joshua D.; Shingler, Kristin L.; Carnegie, Michael S.; Yoder, Jennifer L.; Ashley, Robert E.; Makhov, Alexander M.; Conway, James F.

2013-01-01

158

Antiviral effects of two Ganoderma lucidum triterpenoids against enterovirus 71 infection.  

PubMed

Enterovirus 71 (EV71) is a major causative agent for hand, foot and mouth disease (HFMD), and fatal neurological and systemic complications in children. However, there is currently no clinical approved antiviral drug available for the prevention and treatment of the viral infection. Here, we evaluated the antiviral activities of two Ganoderma lucidum triterpenoids (GLTs), Lanosta-7,9(11),24-trien-3-one,15;26-dihydroxy (GLTA) and Ganoderic acid Y (GLTB), against EV71 infection. The results showed that the two natural compounds display significant anti-EV71 activities without cytotoxicity in human rhabdomyosarcoma (RD) cells as evaluated by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell proliferation assay. The mechanisms by which the two compounds affect EV71 infection were further elucidated by three action modes using Ribavirin, a common antiviral drug, as a positive control. The results suggested that GLTA and GLTB prevent EV71 infection through interacting with the viral particle to block the adsorption of virus to the cells. In addition, the interactions between EV71 virion and the compounds were predicated by computer molecular docking, which illustrated that GLTA and GLTB may bind to the viral capsid protein at a hydrophobic pocket (F site), and thus may block uncoating of EV71. Moreover, we demonstrated that GLTA and GLTB significantly inhibit the replication of the viral RNA (vRNA) of EV71 replication through blocking EV71 uncoating. Thus, GLTA and GLTB may represent two potential therapeutic agents to control and treat EV71 infection. PMID:24845570

Zhang, Wenjing; Tao, Junyan; Yang, Xiaoping; Yang, Zhuliang; Zhang, Li; Liu, Hongsheng; Wu, Kailang; Wu, Jianguo

2014-07-01

159

Mutations in the non-structural protein region contribute to intra-genotypic evolution of enterovirus 71  

PubMed Central

Background Clinical manifestations of enterovirus 71 (EV71) range from herpangina, hand-foot-and-mouth disease (HFMD), to severe neurological complications. Unlike the situation of switching genotypes seen in EV71 outbreaks during 19982008 in Taiwan, genotype B5 was responsible for two large outbreaks in 2008 and 2012, respectively. In China, by contrast, EV71 often persists as a single genotype in the population and causes frequent outbreaks. To investigate genetic changes in viral evolution, complete EV71 genome sequences were used to analyze the intra-genotypic evolution pattern in Taiwan, China, and the Netherlands. Results Genotype B5 was predominant in Taiwans 2008 outbreak and was re-emergent in 2012. EV71 strains from both outbreaks were phylogenetically segregated into two lineages containing fourteen non-synonymous substitutions predominantly in the non-structural protein coding region. In China, genotype C4 was first seen in 1998 and caused the latest large outbreak in 2008. Unlike shifting genotypes in Taiwan, genotype C4 persisted with progressive drift through time. A majority of non-synonymous mutations occurred in residues located in the non-structural coding region, showing annual increases. Interestingly, genotype B1/B2 in the Netherlands showed another stepwise evolution with dramatic EV71 activity increase in 1986. Phylogeny of the VP1 coding region in 19711986 exhibited similar lineage turnover with genotype C4 in China; however, phylogeny of the 3D-encoding region indicated separate lineage appearing after 1983, suggesting that the 3D-encoding region of genotype B2 was derived from an unidentified ancestor that contributed to intra-genotypic evolution in the Netherlands. Conclusions Unlike VP1 coding sequences long used for phylogenetic study of enteroviruses due to expected host immune escape, our study emphasizes a dominant role of non-synonymous mutations in non-structural protein regions that contribute to (re-)emergent genotypes in continuous stepwise evolution. Dozens of amino acid substitutions, especially in non-structural proteins, were identified via genetic changes driven through intra-genotypic evolution worldwide. These identified substitutions appeared to increase viral fitness in the population, affording valuable insights not only for viral evolution but also for prevention, control, and vaccine against EV71 infection. PMID:24766641

2014-01-01

160

[Expression and activity analysis of Enterovirus 71 3C protease in Escherichia coli].  

PubMed

The recombinant plasmid carrying the gene encoding 3C protease of Enterovirus 71 (EV71) was constructed, the recombinant protein was then expressed and purified, the functional activity was also measured. Firstly, the 3C protease gene was inserted into pET28a vector, the constructed recombinant plasmid was transformed into E. coli BL21 (DE3) for expression under the induction of IPTG. The expressed protein was purified by affinity chromatography (Ni-NTA) and the N-terminus His-tag was cleaved by enterokinase from 3C protease. The activity of 3C protease was evaluated with fluorescent peptide substrates. It was verified by restriction analysis and sequencing that recombinant plasmid pET28a-3C was constructed correctly and functionally expressed in E. coli BL21 (DE3) resulting in the production of recombinant 3C protease with a size of 22kD. Both His-tag and non-His-tag (cleaved by enterokinase) 3C protease exhibited similar enzyme activity to 3B-3C fluorescent peptide with Km, Vmax and Kcat values of 22 microM, 434nM. Min(-1) and 0.0669 Min(-1), respectively. The optimial pH and temperature were 7.0 and 30-37 degrees C, respectively. The acquirement of recombinant purified 3C protease with high activity has paved the way of further studies on anti-viral inhibitors, structural protein assembly, vaccine development and detection methods of EV71. PMID:22764519

Chen, Li; Yang, Zhi-Jian; Zhou, Zheng; Cai, Wei-Te; Teng, Xin-Ze; Zhang, Gao-Xia

2012-05-01

161

Development of a quantitative method for the detection of enteroviruses in soil.  

PubMed Central

A method is described for efficiently concentrating enteroviruses from soil. Viruses were eluted from soil by mechanical agitation in high pH glycine buffer containing ethylenediaminetetraacetic acid. The eluted viruses were concentrated on a floc that formed de novo upon adjustment of the soil eluate to 0.06 M aluminum chloride and pH 3.5. Viruses not pelleted with the floc were concentrated by adsorption to and elution from membrane filters. This method yielded an average efficiency of 66% recovery from loamy sand soil for four enteroviruses. Virus recovery from soil was consistently high, with samples ranging in size from 25 to 500 g. The method was used successfully to isolate naturally occurring viruses from soil beneath a wastewater land treatment site. Recovery of enteroviruses by this method form different types of soil was dependent on percentage of clay, surface area, and cation exchange capacity. Recovery was not dependent on soil saturation pH or on percentage of organic matter. This method should prove useful for studying enterovirus migration and survival during the land application of domestic sewage. PMID:36845

Hurst, C J; Gerba, C P

1979-01-01

162

Co-Circulation and Evolution of Polioviruses and Species C Enteroviruses in a District of Madagascar  

Microsoft Academic Search

Between October 2001 and April 2002, five cases of acute flaccid paralysis (AFP) associated with type 2 vaccine-derived polioviruses (VDPVs) were reported in the southern province of the Republic of Madagascar. To determine viral factors that favor the emergence of these pathogenic VDPVs, we analyzed in detail their genomic and phenotypic characteristics and compared them with co-circulating enteroviruses. These VDPVs

Mala Rakoto-Andrianarivelo; Sophie Guillot; Jane Iber; Jean Balanant; Bruno Blondel; Franck Riquet; Javier Martin; Olen Kew; Bakolalao Randriamanalina; Lalatiana Razafinimpiasa; Dominique Rousset; Francis Delpeyroux

2007-01-01

163

Enterovirus, Cytomegalovirus, and Epstein-Barr Virus Infection Screening in Idiopathic Sudden Sensorineural Hearing Loss  

Microsoft Academic Search

Sudden sensorineural hearing loss (SSNHL) is frequently classified as idiopathic since the causative factor is not identified in most cases. In the present study we determined whether SSNHL is associated with common viral infections, namely enterovirus, cytomegalovirus (CMV) and Epstein-Barr virus (EBV). Between April 2004 and March 2005, we conducted a prospective cohort study on 48 unselected patients with unilateral

Menachem Gross; Dana G. Wolf; Josef Elidan; Ron Eliashar

2007-01-01

164

Sequence analysis of a porcine enterovirus serotype 1 isolate: relationships with other picornaviruses  

Microsoft Academic Search

The majority of the genomic sequence of a porcine enterovirus serotype 1 (PEV-1) isolate was determined. The genome was found to contain a large open reading frame which encoded a leader protein prior to the capsid protein region. This showed no sequence identity to other picornavirus leader regions and the sequence data suggested that it does not possess proteolytic activity.

Michelle Doherty; Daniel Todd; Neil McFerran; Elizabeth M. Hoey

1999-01-01

165

Salvage treatment with amphotericin B in progressive human alveolar echinococcosis.  

PubMed

Most patients with alveolar echinococcosis are diagnosed at a late stage when the disease has advanced to unresectable hepatic lesions. These patients require lifelong therapy with benzimidazoles, the only medical treatment currently available. To date, no treatment option remains for patients with benzimidazole intolerance or treatment failure. Amphotericin B was recently shown to exert antiparasitic activity in vitro. Here, we report the efficacy of amphotericin B in human alveolar echinococcosis. In three patients with extensive disease and without further treatment options, disease progression had been documented over several months. They were treated with amphotericin B intravenously at a dose of 0.5 mg/kg of body weight three times per week. Follow-up parameters were physical examination, laboratory parameters, and imaging techniques. Amphotericin B treatment effectively halted parasite growth in all three patients. The antiparasitic effect was most evident by spontaneous closure of cutaneous fistulae in two patients and by constant size of parasitic lesions during treatment, as assessed radiologically. Metabolic activity in parasitic areas was visualized by positron emission tomography and significantly decreased during treatment. However, progressive affection of the heart in one patient could not be stopped. All patients currently continue on amphotericin B and have been treated for 25, 17, and 14 months, respectively. We introduce amphotericin B as salvage treatment for alveolar echinococcosis patients with intolerance or resistance to benzimidazoles, as it effectively suppresses parasite growth. Amphotericin B is not parasitocidal; therefore long-term treatment has to be anticipated. PMID:14576122

Reuter, Stefan; Buck, Andreas; Grebe, Olaf; Nssle-Kgele, Karin; Kern, Peter; Manfras, Burkhard J

2003-11-01

166

Salvage Treatment with Amphotericin B in Progressive Human Alveolar Echinococcosis  

PubMed Central

Most patients with alveolar echinococcosis are diagnosed at a late stage when the disease has advanced to unresectable hepatic lesions. These patients require lifelong therapy with benzimidazoles, the only medical treatment currently available. To date, no treatment option remains for patients with benzimidazole intolerance or treatment failure. Amphotericin B was recently shown to exert antiparasitic activity in vitro. Here, we report the efficacy of amphotericin B in human alveolar echinococcosis. In three patients with extensive disease and without further treatment options, disease progression had been documented over several months. They were treated with amphotericin B intravenously at a dose of 0.5 mg/kg of body weight three times per week. Follow-up parameters were physical examination, laboratory parameters, and imaging techniques. Amphotericin B treatment effectively halted parasite growth in all three patients. The antiparasitic effect was most evident by spontaneous closure of cutaneous fistulae in two patients and by constant size of parasitic lesions during treatment, as assessed radiologically. Metabolic activity in parasitic areas was visualized by positron emission tomography and significantly decreased during treatment. However, progressive affection of the heart in one patient could not be stopped. All patients currently continue on amphotericin B and have been treated for 25, 17, and 14 months, respectively. We introduce amphotericin B as salvage treatment for alveolar echinococcosis patients with intolerance or resistance to benzimidazoles, as it effectively suppresses parasite growth. Amphotericin B is not parasitocidal; therefore long-term treatment has to be anticipated. PMID:14576122

Reuter, Stefan; Buck, Andreas; Grebe, Olaf; Nussle-Kugele, Karin; Kern, Peter; Manfras, Burkhard J.

2003-01-01

167

Isolation and structural organization of the human preproenkephalin B gene  

Microsoft Academic Search

The primary structure of porcine preproenkephalin B has been elucidated by cloning and sequencing cDNA1: it contains neoendorphin2,3, dynorphin4,5 and leumorphin1,6,7 (containing rimorphin as its ammo-terminus)8,9. These opioid peptides, each having a leucine-enkephalin structure10, act on the kappa-receptor7,11-14. We have now cloned a human genomic DNA segment containing the preproenkephalin B gene. The structural organization of this gene resembles those

Saburo Horikawa; Toshiyuki Takai; Mitsuyoshi Toyosato; Hideo Takahashi; Masaharu Noda; Hitoshi Kakidani; Tai Kubo; Tadaaki Hirose; Seiichi Inayama; Hidenori Hayashida; Takashi Miyata; Shosaku Numa

1983-01-01

168

Activation of human B-MYB by cyclins  

PubMed Central

B-MYB expression is associated with cell proliferation and recent studies have suggested that it promotes the S phase of mammalian cells. Based on its homology to the transcription factors c-MYB and A-MYB, B-MYB is thought to be involved in transcriptional regulation; however, its activity is not detectable in several cell lines. It was postulated that B-MYB function may depend on the presence of a cofactor, and recent studies suggested that B-MYB is phosphorylated specifically during S phase in murine fibroblasts. In this report we provide evidence that the product of the human B-myb gene can be activated in vivo by coexpression with cyclin A or cyclin E. Transfection studies showed that B-MYB was a weak transcriptional activator in SAOS-2 cells and was unable to promote their proliferation. In contrast, overexpression of both B-MYB and cyclin A or cyclin E caused a drastic increase in the number of SAOS-2 cells in S phase. Also, overexpression of cyclin A and cyclin E in SAOS-2 cells enhanced the ability of B-MYB, but not c-MYB, to transactivate various promoters, including the cdc2 promoter, the HIV-1-LTR, and the simian virus 40 minimal promoter. A direct role for cyclin-dependent activation of B-MYB was demonstrated using an in vitro transcription assay. These observations suggest that one mechanism by which cyclin A and E may promote the S phase is through modification and activation of B-MYB. PMID:9012818

Sala, Arturo; Kundu, Mondira; Casella, Ida; Engelhard, Andrew; Calabretta, Bruno; Grasso, Luigi; Paggi, Marco G.; Giordano, Antonio; Watson, Roger J.; Khalili, Kamel; Peschle, Cesare

1997-01-01

169

Human Papillomavirus Type 31b Infection of Human Keratinocytes and the Onset of Early Transcription  

PubMed Central

Human papillomaviruses (HPVs) cause a number of human tumors and malignancies, including cervical cancers. Epithelial differentiation is required for the complete HPV life cycle and can be achieved using the organotypic (raft) culture system. The CIN-612 9E cell line maintains episomal copies of HPV type 31b (HPV31b), an HPV type associated with cervical cancers. When grown in the raft system, CIN-612 9E cells form a differentiated epithelium such that infectious virions can be synthesized. Many aspects of the later stages of the HPV31b life cycle have been investigated in CIN-612 9E raft tissues. We used a biologically contained homogenization system for efficient virion extraction from raft epithelial tissues. Purified HPV31b virions were used to infect low-passage-number human foreskin keratinocytes and a variety of epithelial cell lines. Newly synthesized, spliced HPV31b transcripts were detected by reverse transcription and PCR (RT-PCR) following HPV31b infection. HPV31b infection was most efficient and reproducible in HaCaT cells. The onset of viral transcription following infection was also investigated using RT-PCR techniques. Spliced E1?I,E2 RNAs were present as early as 4 h postinfection (p.i.), whereas the other major viral transcripts were detected by 8 to 10 h p.i. Furthermore, we characterized the structures and temporal expression of seven novel spliced early transcripts expressed following infection. PMID:12388689

Ozbun, Michelle A.

2002-01-01

170

Adenosine production by human B cells and B cell-mediated suppression of activated T cells  

PubMed Central

Antibody-independent role of B cells in modulating T-cell responses is incompletely understood. Freshly isolated or cultured B cells isolated from the peripheral blood of 30 normal donors were evaluated for CD39 and CD73 coexpression, the ability to produce adenosine 5?-monophosphate (AMP) and adenosine (ADO) in the presence of exogenous adenosine triphosphate (ATP) as well as A1, A2A, A2B, and A3 adenosine receptor (ADOR) expression. Human circulating B cells coexpress ectonucleotidases CD39 and CD73, hydrolyze exogenous ATP to 5?-AMP and ADO, and express messenger RNA for A1R, A2AR, and A3R. 2-chloroadenosine inhibited B-cell proliferation and cytokine expression, and only A3R selective antagonist restored B-cell functions. This suggested that B cells use the A3R for autocrine signaling and self-regulation. Mediated effects on B-cell growth ADOR antagonists or agonists were tested in carboxyfluorescein diacetate succinimidyl ester assays. In cocultures, resting B cells upregulated functions of CD4+ and CD8+ T cells. However, in vitroactivated B cells downregulated CD73 expression, mainly produced 5?-AMP, and inhibited T-cell proliferation and cytokine production. These B cells acquire the ability to restrict potentially harmful effects of activated T cells. Thus, B cells emerge as a key regulatory component of T cellB cell interactions, and their dual regulatory activity is mediated by the products of ATP hydrolysis, 5?-AMP, and ADO. PMID:23678003

Saze, Zenichiro; Schuler, Patrick J.; Hong, Chang-Sook; Cheng, Dongmei; Jackson, Edwin K.

2013-01-01

171

Human parechoviruses: Biology, epidemiology and clinical significance  

Microsoft Academic Search

Human parechoviruses (HPeVs) are members of the large and growing family of Picornaviridae. Although originally described as echovirus 22 and 23 within human enteroviruses because of their clinical and morphological properties, they have since been shown to be distinct from this and other picornavirus groups in several features of their genome organisation, structure and replication. Human parechoviruses show genetic and

H. Harvala; P. Simmonds

2009-01-01

172

Histone demethylases KDM4B and KDM6B promotes osteogenic differentiation of human MSCs.  

PubMed

Human bone marrow mesenchymal stem/stromal cells (MSCs) are multipotent progenitor cells with multilineage differentiation potentials including osteogenesis and adipogenesis. While significant progress has been made in understanding transcriptional controls of MSC fate, little is known about how MSC differentiation is epigenetically regulated. Here we show that the histone demethylases KDM4B and KDM6B play critical roles in osteogenic commitment of MSCs by removing H3K9me3 and H3K27me3. Depletion of KDM4B or KDM6B significantly reduced osteogenic differentiation and increased adipogenic differentiation. Mechanistically, while KDM6B controlled HOX expression by removing H3K27me3, KDM4B promoted DLX expression by removing H3K9me3. Importantly, H3K27me3- and H3K9me3-positive MSCs of bone marrow were significantly elevated in ovariectomized and aging mice in which adipogenesis was highly active. Since histone demethylases are chemically modifiable, KDM4B and KDM6B may present as therapeutic targets for controlling MSC fate choices and lead to clues for newtreatment in metabolic bone diseases such as osteoporosis. PMID:22770241

Ye, Ling; Fan, Zhipeng; Yu, Bo; Chang, Jia; Al Hezaimi, Khalid; Zhou, Xuedong; Park, No-Hee; Wang, Cun-Yu

2012-07-01

173

Comparing Enterovirus 71 with Coxsackievirus A16 by analyzing nucleotide sequences and antigenicity of recombinant proteins of VP1s and VP4s  

PubMed Central

Background Enterovirus 71 (EV71) and Coxsackievirus A16 (CA16) are two major etiological agents of Hand, Foot and Mouth Disease (HFMD). EV71 is associated with severe cases but not CA16. The mechanisms contributed to the different pathogenesis of these two viruses are unknown. VP1 and VP4 are two major structural proteins of these viruses, and should be paid close attention to. Results The sequences of vp1s from 14 EV71 and 14 CA16, and vp4s from 10 EV71 and 1 CA16 isolated in this study during 2007 to 2009 HFMD seasons were analyzed together with the corresponding sequences available in GenBank using DNAStar and MEGA 4.0. Phylogenetic analysis of complete vp1s or vp4s showed that EV71 isolated in Beijing belonged to C4 and CA16 belonged to lineage B2 (lineage C). VP1s and VP4s from 4 strains of viruses expressed in E. coli BL21 cells were used to detect IgM and IgG in human sera by Western Blot. The detection of IgM against VP1s of EV71 and CA16 showed consistent results with current infection, while none of the sera were positive against VP4s of EV71 and CA16. There was significant difference in the positive rates between EV71 VP1 and CA16 VP1 (?2 = 5.02, P < 0.05) as well as EV71 VP4 and CA16 VP4 (?2 = 15.30, P < 0.01) in the detection of IgG against recombinant proteins with same batch of serum samples. The sera-positive rate of IgG against VP1 was higher than that against VP4 for both EV71 (?2 = 26.47, P < 0.01) and CA16 (?2 = 16.78, P < 0.01), which might be because of different positions of VP1 and VP4 in the capsid of the viruses. Conclusions EV71 and CA16 were highly diverse in the nucleotide sequences of vp1s and vp4s. The sera positive rates of VP1 and VP4 of EV71 were lower than those of CA16 respectively, which suggested a less exposure rate to EV71 than CA16 in Beijing population. Human serum antibodies detected by Western blot using VP1s and VP4s as antigen indicated that the immunological reaction to VP1 and VP4 of both EV71 and CA16 was different. PMID:22050722

2011-01-01

174

Recruitment of PI4KIII? to Coxsackievirus B3 Replication Organelles Is Independent of ACBD3, GBF1, and Arf1  

PubMed Central

ABSTRACT Members of the Enterovirus (poliovirus [PV], coxsackieviruses, and human rhinoviruses) and Kobuvirus (Aichi virus) genera in the Picornaviridae family rely on PI4KIII? (phosphatidylinositol-4-kinase III?) for efficient replication. The small membrane-anchored enteroviral protein 3A recruits PI4KIII? to replication organelles, yet the underlying mechanism has remained elusive. Recently, it was shown that kobuviruses recruit PI4KIII? through interaction with ACBD3 (acyl coenzyme A [acyl-CoA]-binding protein domain 3), a novel interaction partner of PI4KIII?. Therefore, we investigated a possible role for ACBD3 in recruiting PI4KIII? to enterovirus replication organelles. Although ACBD3 interacted directly with coxsackievirus B3 (CVB3) 3A, its depletion from cells by RNA interference did not affect PI4KIII? recruitment to replication organelles and did not impair CVB3 RNA replication. Enterovirus 3A was previously also proposed to recruit PI4KIII? via GBF1/Arf1, based on the known interaction of 3A with GBF1, an important regulator of secretory pathway transport and a guanine nucleotide exchange factor (GEF) of Arf1. However, our results demonstrate that inhibition of GBF1 or Arf1 either by pharmacological inhibition or depletion with small interfering RNA (siRNA) treatment did not affect the ability of 3A to recruit PI4KIII?. Furthermore, we show that a 3A mutant that no longer binds GBF1 was capable of recruiting PI4KIII?, even in ACBD3-depleted cells. Together, our findings indicate that unlike originally envisaged, coxsackievirus recruits PI4KIII? to replication organelles independently of ACBD3 and GBF1/Arf1. IMPORTANCE A hallmark of enteroviral infection is the generation of new membranous structures to support viral RNA replication. The functionality of these replication organelles depends on the concerted actions of both viral nonstructural proteins and co-opted host factors. It is thus essential to understand how these structures are formed and which cellular components are key players in this process. GBF1/Arf1 and ACBD3 have been proposed to contribute to the recruitment of the essential lipid-modifying enzyme PI4KIII? to enterovirus replication organelles. Here we show that the enterovirus CVB3 recruits PI4KIII? by a mechanism independent of both GBF1/Arf1 and ACBD3. This study shows that the strategy employed by coxsackievirus to recruit PI4KIII? to replication organelles is far more complex than initially anticipated. PMID:24352456

Dorobantu, Cristina M.; van der Schaar, Hilde M.; Ford, Lauren A.; Strating, Jeroen R. P. M.; Ulferts, Rachel; Fang, Ying; Belov, George

2014-01-01

175

An RNA replication-center assay for high content image-based quantifications of human rhinovirus and coxsackievirus infections  

PubMed Central

Background Picornaviruses are common human and animal pathogens, including polio and rhinoviruses of the enterovirus family, and hepatits A or food-and-mouth disease viruses. There are no effective countermeasures against the vast majority of picornaviruses, with the exception of polio and hepatitis A vaccines. Human rhinoviruses (HRV) are the most prevalent picornaviruses comprising more than one hundred serotypes. The existing and also emerging HRVs pose severe health risks for patients with asthma or chronic obstructive pulmonary disease. Here, we developed a serotype-independent infection assay using a commercially available mouse monoclonal antibody (mabJ2) detecting double-strand RNA. Results Immunocytochemical staining for RNA replication centers using mabJ2 identified cells that were infected with either HRV1A, 2, 14, 16, 37 or coxsackievirus (CV) B3, B4 or A21. MabJ2 labeled-cells were immunocytochemically positive for newly synthesized viral capsid proteins from HRV1A, 14, 16, 37 or CVB3, 4. We optimized the procedure for detection of virus replication in settings for high content screening with automated fluorescence microscopy and single cell analysis. Our data show that the infection signal was dependent on multiplicity, time and temperature of infection, and the mabJ2-positive cell numbers correlated with viral titres determined in single step growth curves. The mabJ2 infection assay was adapted to determine the efficacy of anti-viral compounds and small interfering RNAs (siRNAs) blocking enterovirus infections. Conclusions We report a broadly applicable, rapid protocol to measure infection of cultured cells with enteroviruses at single cell resolution. This assay can be applied to a wide range of plus-sense RNA viruses, and hence allows comparative studies of viral infection biology without dedicated reagents or procedures. This protocol also allows to directly compare results from small compound or siRNA infection screens for different serotypes without the risk of assay specific artifacts. PMID:20937137

2010-01-01

176

Human Rhinoviruses  

PubMed Central

Human rhinoviruses (HRVs), first discovered in the 1950s, are responsible for more than one-half of cold-like illnesses and cost billions of dollars annually in medical visits and missed days of work. Advances in molecular methods have enhanced our understanding of the genomic structure of HRV and have led to the characterization of three genetically distinct HRV groups, designated groups A, B, and C, within the genus Enterovirus and the family Picornaviridae. HRVs are traditionally associated with upper respiratory tract infection, otitis media, and sinusitis. In recent years, the increasing implementation of PCR assays for respiratory virus detection in clinical laboratories has facilitated the recognition of HRV as a lower respiratory tract pathogen, particularly in patients with asthma, infants, elderly patients, and immunocompromised hosts. Cultured isolates of HRV remain important for studies of viral characteristics and disease pathogenesis. Indeed, whether the clinical manifestations of HRV are related directly to viral pathogenicity or secondary to the host immune response is the subject of ongoing research. There are currently no approved antiviral therapies for HRVs, and treatment remains primarily supportive. This review provides a comprehensive, up-to-date assessment of the basic virology, pathogenesis, clinical epidemiology, and laboratory features of and treatment and prevention strategies for HRVs. PMID:23297263

Lamson, Daryl M.; St. George, Kirsten; Walsh, Thomas J.

2013-01-01

177

The variations of VP1 protein might be associated with nervous system symptoms caused by enterovirus 71 infection  

PubMed Central

Background The VP1 protein of enterovirus 71 (EV71) is an important immunodominant protein which is responsible for host-receptor binding. Nevertheless, the relationship between VP1 and neurovirulence is still poorly understood. In this study, we investigated the relationship between mutation of VP1 and neurovirulent phenotype of EV71 infection. Methods One hundred and eighty-seven strains from Genbank were included, with a clear clinical background. They were divided into two groups, one with nervous system symptoms and one with no nervous system symptoms. After alignment, the significance of amino acid variation was determined by using the ?2 test and a phylogenetic tree was constructed with MEGA software (version 5.1). Results We showed no significant difference in neurovirulence between genotype B and C. Interestingly, we found that variations of E145G/Q, E164D/K and T292N/K were associated with nervous system infection in genotype B. In the case of genotype C, the N31D mutation increased the risk for nervous complications, whereas I262V mutation decreased the risk of nervous complications. We used a 3D model of VP1 to demonstrate the potential molecular basis for EV71 nervous system tropism. Conclusions Distinct variations are shown to be associated with neurovirulent phenotype in the different genotype. Detection of variation in genotypes and subtypes may be important for the prediction of clinical outcomes. PMID:24886383

2014-01-01

178

Evidence for APOBEC3B mutagenesis in multiple human cancers  

PubMed Central

Thousands of somatic mutations accrue in most human cancers and causes are largely unknown. We recently showed that the DNA cytosine deaminase APOBEC3B accounts for up to half of the mutational load in breast carcinomas expressing this enzyme. Here, we address whether APOBEC3B is broadly responsible for mutagenesis in multiple tumor types. We analyzed gene expression data and mutation patterns, distributions, and loads for 19 different cancer types, totaling over 4,800 exomes and 1,000,000 somatic mutations. Remarkably, APOBEC3B is upregulated and its preferred target sequence is frequently mutated and clustered in at least 6 distinct cancers: bladder, cervix, lung (adeno- and squamous cell), head/neck, and breast. Interpreted in light of prior genetic, cellular, and biochemical studies, the most parsimonious conclusion based on these global analyses is that APOBEC3B catalyzed genomic uracil lesions are responsible for a large proportion of both dispersed and clustered mutations in multiple distinct cancers. PMID:23852168

Burns, Michael B.; Temiz, Nuri A.; Harris, Reuben S.

2014-01-01

179

Human Health Countermeasures (HHC) Element Management Plan: Human Research Program. Revision B  

NASA Technical Reports Server (NTRS)

NASA s Human Research Program (HRP) is an applied research and technology program within the Human Exploration and Operations Mission Directorate (HEOMD) that addresses human health and performance risk mitigation strategies in support of exploration missions. The HRP research and technology development is focused on the highest priority risks to crew health and safety with the goal of ensuring mission success and maintaining long-term crew health. Crew health and performance standards, defined by the NASA Chief Health and Medical Officer (CHMO), set the acceptable risk level for exploration missions. The HRP conducts research to inform these standards as well as provide deliverables, such as countermeasures, that ensure standards can be met to maximize human performance and mission success. The Human Health Countermeasures (HHC) Element was formed as part of the HRP to develop a scientifically-based, integrated approach to understanding and mitigating the health risks associated with human spaceflight. These health risks have been organized into four research portfolios that group similar or related risks. A fifth portfolio exists for managing technology developments and infrastructure projects. The HHC Element portfolios consist of: a) Vision and Cardiovascular; b) Exercise and Performance; c) Multisystem; d) Bone; and e) Technology and Infrastructure. The HHC identifies gaps associated with the health risks and plans human physiology research that will result in knowledge required to more fully understand risks and will result in validated countermeasures to mitigate risks.

Norsk, Peter; Baumann, David

2012-01-01

180

Purinoceptor-operated cationic channels in human B lymphocytes.  

PubMed Central

1. Using the patch clamp method in the outside-out configuration, purinoceptor-dependent unitary currents were measured in tonsillar and transformed tonsillar human B lymphocytes. 2. Single channel currents were evoked by ATP4-, the free-acid form of ATP, and by 2',3' O-benzoyl-4-benzoyl-ATP (BzATP) in the micromolar concentration range, but not by 10 mM ADP3- or 0.5 mM Mg(2+)-bound ATP. 3. The channels could be activated and deactivated several times for as long as 30 min even in the absence of intracellular ATP, GTP, or glucose. 4. The channels were selective for small cations and had a conductance of 9 pS with Cs+ as the intracellular and Na+ as the extracellular monovalent cation. 5. The half-maximal activation of the channels was obtained by 114 microM ATP4- and by 16 microM BzATP. The increase in the open probability after raising the ATP4- concentration was mainly due to a decrease in the times the channels spend in the closed state. 6. It is concluded that human B lymphocytes possess cationic channels directly gated by extracellular ATP4-. Their agonist binding characteristics are typical for P2z purinoceptors, but their permeation behaviour is different from the large non-specific pores formed by ATP4- in fibroblasts, macrophages and mast cells. PMID:9023774

Markwardt, F; Lohn, M; Bohm, T; Klapperstuck, M

1997-01-01

181

Diagnosis of horizontal enterovirus infections in neonates by nested PCR and direct sequence analysis  

Microsoft Academic Search

A hospital-acquired outbreak with febrile illness and\\/or rash occurred in our neonatal special care nursery (SCN) from September 1995 to September 1996. A total of 23 infants developed symptoms. We could not detect the etiological agents by routine virus isolation. In a retrospective study, however, enterovirus RNA was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) from four out of six

T Takami; S Sonoda; H Houjyo; H Kawashima; Y Takei; T Miyajima; K Takekuma; A Hoshika; T Mori; T Nakayama

2000-01-01

182

Comparison of Automated and Manual Nucleic Acid Extraction Methods for Detection of Enterovirus RNA  

Microsoft Academic Search

Automated nucleic acid extraction is an attractive alternative to labor-intensive manual methods. We compared two automated methods, the BioRobot M48 instrument (Qiagen, Inc.) and MagNA Pure (Roche Applied Sciences) methods, to two manual methods, the QIAamp Viral RNA Mini kit (Qiagen) and TRIzol (Invitrogen), for the extraction of enterovirus RNA. Analytical sensitivity was assessed by dilution analysis of poliovirus type

Julia H. Knepp; Melissa A. Geahr; Michael S. Forman; Alexandra Valsamakis

2003-01-01

183

Human Monoclonal Antibodies Broadly Neutralizing against Influenza B Virus  

PubMed Central

Influenza virus has the ability to evade host immune surveillance through rapid viral genetic drift and reassortment; therefore, it remains a continuous public health threat. The development of vaccines producing broadly reactive antibodies, as well as therapeutic strategies using human neutralizing monoclonal antibodies (HuMAbs) with global reactivity, has been gathering great interest recently. Here, three hybridoma clones producing HuMAbs against influenza B virus, designated 5A7, 3A2 and 10C4, were prepared using peripheral lymphocytes from vaccinated volunteers, and were investigated for broad cross-reactive neutralizing activity. Of these HuMAbs, 3A2 and 10C4, which recognize the readily mutable 190-helix region near the receptor binding site in the hemagglutinin (HA) protein, react only with the Yamagata lineage of influenza B virus. By contrast, HuMAb 5A7 broadly neutralizes influenza B strains that were isolated from 1985 to 2006, belonging to both Yamagata and Victoria lineages. Epitope mapping revealed that 5A7 recognizes 316G, 318C and 321W near the C terminal of HA1, a highly conserved region in influenza B virus. Indeed, no mutations in the amino acid residues of the epitope region were induced, even after the virus was passaged ten times in the presence of HuMAb 5A7. Moreover, 5A7 showed significant therapeutic efficacy in mice, even when it was administered 72 hours post-infection. These results indicate that 5A7 is a promising candidate for developing therapeutics, and provide insight for the development of a universal vaccine against influenza B virus. PMID:23408886

Yasugi, Mayo; Kubota-Koketsu, Ritsuko; Yamashita, Akifumi; Kawashita, Norihito; Du, Anariwa; Sasaki, Tadahiro; Nishimura, Mitsuhiro; Misaki, Ryo; Kuhara, Motoki; Boonsathorn, Naphatsawan; Fujiyama, Kazuhito; Okuno, Yoshinobu; Nakaya, Takaaki; Ikuta, Kazuyoshi

2013-01-01

184

Survival of enteroviruses in rapid-infiltration basins during the land application of wastewater.  

PubMed

The downward migration through soil of seeded poliovirus type 1 and echovirus type 1 and of naturally occurring enteroviruses during infiltration of sewage effluent through rapid-infiltration basins was investigated. After 5 days of flooding, the amount of seeded poliovirus type 1 that had migrated 5 to 10 cm downward through the soil profile was found to be 11% of that remaining at the initial burial depth. The amount of echovirus type 1 determined to have moved an equal distance was at least 100-fold less. Migration of naturally occurring enteroviruses during infiltration of sewage effluent through soil could not be measured with accuracy because of the possibility of virus survival from previous applications of effluent. The rate of inactivation for seeded poliovirus 1 and echovirus 1 buried in the infiltration basins ranged between 0.04 and 0.15 log10 units per day during the time when the basins were flooded. Inactivation of these same seeded virus types and of indigenous enterovirus populations in the infiltration basins during the drying portion of the sewage application cycle ranged between 0.11 and 0.52 log10 units per day. The rate of virus inactivation was dependent upon the rate of soil moisture loss. These results indicate that drying cycles during the land application of wastewater enhance virus inactivation in the soil. PMID:6258471

Hurst, C J; Gerba, C P; Lance, J C; Rice, R C

1980-08-01

185

A comparison of two extraction methods for the detection of Enteroviruses in raw sludge.  

PubMed

The aim of this study was to compare two viral extraction methods for the detection of naturally occurring Enteroviruses in raw sludge. The first method (M1) is based on an ultracentrifugation step. In the second one (M2), viral RNA was extracted directly after viral elution from suspended solids. Genomes of enteroviruses were quantified by a quantitative real time PCR (qRT-PCR) in sludge samples. Somatic coliphages and F-specific RNA phages, considered as viral indicators of enteric viruses in sludge, were enumerated by the double layer agar technique. Results showed that direct assay of RNA extraction yielded higher genomic copies of enteric viruses (with an average of 5.07Log10 genomic copies/100 mL). After the ultracentrifugation assay in the second method, genomic copies number decreases (with an average of 4.39Log10 genomic copies/100 mL). This can be explained by an eventual concentration of inhibitors existing in sludge samples. Phages enumeration results showed their presence in all sludge samples with an average of (5.69Log10 PFU/100 mL) for somatic coliphages and (4Log10 PFU/100 mL) for F-specific RNA phages. This emphasizes the use of somatic coliphages as viral indicators for enteroviruses in environmental samples and especially in raw sludge samples in wastewater treatment plants prior to agricultural use. PMID:24503039

Jebri, Sihem; Hmaied, Fatma; Lucena, Francisco; Saavedra, Mari Eugeni; Yahya, Mariem; Hamdi, Moktar

2014-05-01

186

Outbreak of vertigo in Wyoming: possible role of an enterovirus infection.  

PubMed

An epidemiologic investigation was conducted to characterize and evaluate the possibility of a viral aetiology of an outbreak of acute vertigo in Hot Springs Country, Wyoming, during autumn 1992. Case-finding identified Hot Springs County residents who sought medical attention for new onset vertigo during 1 August, 1992-31 January 1993. Thirty-five case-patients and 61 matched controls were interviewed and serum specimens were obtained during January 1993. Case-patients were more likely than controls to report symptoms (e.g. fatigue, sore throat, fever, diarrhoea) of antecedent acute illness. Case-patients did not have a significantly greater prevalence or mean titre of IgG antibodies to respiratory syncytial virus, parainfluenza viruses, Epstein-Barr virus, and cytomegalovirus than controls. Serologic evidence of recent enterovirus infection (IgM antibodies) was found for 74% of case-patients compared with 54% of controls (P < 0.05), suggesting a possible association between vertigo and enterovirus infection. Future studies are needed to define the role of enteroviruses in innerear diseases. PMID:8760963

Simonsen, L; Khan, A S; Gary, H E; Hanson, C; Pallansch, M A; Music, S; Holman, R C; Stewart, J A; Erdman, D D; Arden, N H; Arenberg, I K; Schonberger, L B

1996-08-01

187

Concentration and purification of enterovirus 71 using a weak anion-exchange monolithic column  

PubMed Central

Background Enterovirus 71 (EV-71) is a neurotropic virus causing Hand, Foot and Mouth Disease (HFMD) in infants and children under the age of five. It is a major concern for public health issues across Asia-Pacific region. The most effective way to control the disease caused by EV-71 is by vaccination thus a novel vaccine is urgently needed. Inactivated EV-71 induces a strong, virus-neutralizing antibody response in animal models, protecting them against a lethal EV-71 challenge and it has been shown to elicit cross-neutralizing antibodies in human trials. Hence, the large-scale production of purified EV-71 is required for vaccine development, diagnosis and clinical trials. Methods CIM Monolith columns are single-piece columns made up of poly(glycidyl methacrylate co-ethylene dimethacrylate) as support matrix. They are designed as porous channels rather than beads with different chemistries for different requirements. As monolithic columns have a high binding capacity, flow rate and resolution, a CIM DEAE-8f tube monolithic column was selected for purification in this study. The EV-71 infected Rhabdomyosarcoma (RD) cell supernatant was concentrated using 8% PEG 8000 in the presence of 400mM sodium chloride. The concentrated virus was purified by weak anion exchange column using 50mM HEPES?+?1M sodium chloride as elution buffer. Results Highly pure viral particles were obtained at a concentration of 350mM sodium chloride as confirmed by SDS-PAGE and electron microscopy. Presence of viral proteins VP1, VP2 and VP3 was validated by western blotting. The overall process achieved a recovery of 55%. Conclusions EV-71 viral particles of up to 95% purity can be recovered by a single step ion-exchange chromatography using CIM-DEAE monolithic columns and 1M sodium chloride as elution buffer. Moreover, this method is scalable to purify several litres of virus-containing supernatant, using industrial monolithic columns with a capacity of up to 8L such as CIM cGMP tube monolithic columns. PMID:24884895

2014-01-01

188

Spatio-temporal analysis on enterovirus cases through integrated surveillance in Taiwan  

PubMed Central

Background Severe epidemics of enterovirus have occurred frequently in Malaysia, Singapore, Taiwan, Cambodia, and China, involving cases of pulmonary edema, hemorrhage and encephalitis, and an effective vaccine has not been available. The specific aim of this study was to understand the epidemiological characteristics of mild and severe enterovirus cases through integrated surveillance data. Methods All enterovirus cases in Taiwan over almost ten years from three main databases, including national notifiable diseases surveillance, sentinel physician surveillance and laboratory surveillance programs from July 1, 1999 to December 31, 2008 were analyzed. The Pearsons correlation coefficient was applied for measuring the consistency of the trends in the cases between different surveillance systems. Cross correlation analysis in a time series model was applied for examining the capability to predict severe enterovirus infections. Poisson temporal, spatial and space-time scan statistics were used for identifying the most likely clusters of severe enterovirus outbreaks. The directional distribution method with two standard deviations of ellipse was applied to measure the size and the movement of the epidemic. Results The secular trend showed that the number of severe EV cases peaked in 2008, and the number of mild EV cases was significantly correlated with that of severe ones occurring in the same week [r?=?0.553, p?

2014-01-01

189

[Non detection of enterovirus in the bivalve Anadara tuberculosa (Bivalvia:Arcidae) caused by chemical contamination in the Pacific of Costa Rica].  

PubMed

Anadara tuberculosa is one of the most abundant mollusks of commercial importance in Costa Rica. Its habitat water is a potential source of fecal and chemical contamination to humans. We wanted to asses enterovirus, mainly poliovirus and hepatitis A virus and chemicals such as sulphates and nitrates in meat and body fluids. Thirteen samples were taken from four sites in Nicoya Gulf, three sites in the Sierpe-Trraba mangrove (Pacific of Costa Rica) and from five fish markets in San Jos, the capital of Costa Rica. Samples were tested for 1) fecal coliforms (Most Probable Number/100 ml), 2) isolation of enterovirus in cell culture (Hep-2, FrhK-4), 3) cell cytotoxicity in Vero cells and 4) the ability to inactivate 10 ID50% of poliovirus in cell culture. The Most Probable Number/100 ml in surrounding water was higher than the accepted standard for recreational waters, although the number of fecal coliforms in meats and body fluids was lower than in the external water. No cytopathogenic agents were isolated, but we found nitrate and sulphate concentrations that exceeded maxima for human consumption and recreation. The intrinsic cytotoxicity of the samples was at a 1/8 dilution, but some samples were cytotoxic at dilutions of 1/128. Body fluids were more cytotoxic than meats, but a positive correlation between cytotoxicity and chemical contamination was not determined: apparently other pollutants not identified in this study were responsible. Fluid and meat capacity to inactivate 10 ID50% of poliovirus in cell culture was demonstrated. Samples that were toxic for cell cultures also showed a higher percentage of poliovirus inactivation. Monitoring chemical pollution in these waters is highly recommended. PMID:10883327

Libia Herrero, U; Alejandro Palacios, F; Laya Hun, O; Francisco Vega, A

1999-09-01

190

Constitutive and Inducible Expression of B7 Family of Ligands by Human Airway Epithelial Cells  

Microsoft Academic Search

Activated T cells have been implicated in chronic rhinosinusitis (CRS) and asthma and physically interact with epithelial cells in the airways. We now report that human airway epithelial cells display significant constitutive cell-surfaceexpression of costimulatory ligands, B7-H1, B7-H2, B7-H3, and B7-DC. Expression of B7-H1 and B7-DC was selectively induced by stimulation of either BEAS2B or primary nasal epithelial cells (PNEC)

Jean Kim; Allen C. Myers; Lieping Chen; Drew M. Pardoll; Quynh-Ai Truong-Tran; John F. McDyer; Lowella Fortuno; Robert P. Schleimer

2005-01-01

191

Molecular Cell Human LSD2/KDM1b/AOF1 Regulates  

E-print Network

Molecular Cell Article Human LSD2/KDM1b/AOF1 Regulates Gene Transcription by Modulating Intragenic to the regulation of transcriptional initiation. We now show that human LSD2/KDM1b/AOF1, the human homolog of LSD1 regions of its target genes. Genome-wide mapping reveals that LSD2 associates predominantly with the gene

Yuan, Guo-Cheng "GC"

192

Human Hepatocellular Carcinoma Cell Lines Secrete the Major Plasma Proteins and Hepatitis B Surface Antigen  

Microsoft Academic Search

Analysis of the cell culture fluid from two new human hepatoma-derived cell lines reveals that 17 of the major human plasma proteins are synthesized and secreted by these cells. One of these cell lines, Hep 3B, also produces the two major polypeptides of the hepatitis B virus surface antigen. When Hep 3B is injected into athymic mice, metastatic hepatocellular carcinomas

Barbara B. Knowles; Chin C. Howe; David P. Aden

1980-01-01

193

Organometallic B12 -DNA Conjugate: Synthesis, Structure Analysis, and Studies of Binding to Human B12 -Transporter Proteins.  

PubMed

Design, synthesis, and structural characterization of a B12 -octadecanucleotide are presented herein, a new organometallic B12 -DNA conjugate. In such covalent conjugates, the natural B12 moiety may be a versatile vector for controlled in vivo delivery of oligonucleotides to cellular targets in humans and animals, through the endogenous B12 transport systems. Binding of the organometallic B12 octadecanucleotide to the three important human proteins of B12 transport was studied, to examine its structural suitability for the task of eventual in vivo oligonucleotide delivery. Binding was efficient with transcobalamin (TC), but not so efficient with the homologous glycoproteins intrinsic factor and haptocorrin. Binding of the B12 octadecanucleotide to TC suggests the capacity of the B12 moiety to serve as a natural vector for specific transport of single stranded, organometallic oligonucleotide loads from the blood stream into cells. PMID:25168390

Hunger, Miriam; Mutti, Elena; Rieder, Alexander; Enders, Barbara; Nexo, Ebba; Krutler, Bernhard

2014-10-01

194

The Human Leukocyte Antigen-presented Ligandome of B Lymphocytes*  

PubMed Central

Peptides presented by human leukocyte antigen (HLA) molecules on the cell surface play a crucial role in adaptive immunology, mediating the communication between T cells and antigen presenting cells. Knowledge of these peptides is of pivotal importance in fundamental studies of T cell action and in cellular immunotherapy and transplantation. In this paper we present the in-depth identification and relative quantification of 14,500 peptide ligands constituting the HLA ligandome of B cells. This large number of identified ligands provides general insight into the presented peptide repertoire and antigen presentation. Our uniquely large set of HLA ligands allowed us to characterize in detail the peptides constituting the ligandome in terms of relative abundance, peptide length distribution, physicochemical properties, binding affinity to the HLA molecule, and presence of post-translational modifications. The presented B-lymphocyte ligandome is shown to be a rich source of information by the presence of minor histocompatibility antigens, virus-derived epitopes, and post-translationally modified HLA ligands, and it can be a good starting point for solving a wealth of specific immunological questions. These HLA ligands can form the basis for reversed immunology approaches to identify T cell epitopes based not on in silico predictions but on the bona fide eluted HLA ligandome. PMID:23481700

Hassan, Chopie; Kester, Michel G. D.; de Ru, Arnoud H.; Hombrink, Pleun; Drijfhout, Jan Wouter; Nijveen, Harm; Leunissen, Jack A. M.; Heemskerk, Mirjam H. M.; Falkenburg, J. H. Frederik; van Veelen, Peter A.

2013-01-01

195

BAFF ENHANCES CHEMOTAXIS OF PRIMARY HUMAN B CELLS. A PARTICULAR SYNERGY BETWEEN BAFF AND CXCL13 ON MEMORY B CELLS  

Microsoft Academic Search

B-cell-activating factor of the TNF family, (BAFF), and a proliferation-inducing li- gand (APRIL) regulate B-lymphocyte sur- vival and activation. We report that BAFF, but not APRIL, increased the chemotactic response of primary human B cells to CCL21, CXCL12, and CXCL13. The BAFF- induced increase in B-cell chemotaxis was totally abolished by blockade of BAFF-R and was strongly dependent on the

Gamal BADR; Gwenoline BORHIS; Eric A. LEFEVRE; Nada CHAOUL; Frederique DESHAYES; Valrie DESSIRIER; Genevieve LAPREE; Andreas TSAPIS; Yolande RICHARD

2008-01-01

196

Chimeric virus-like particle vaccines displaying conserved enterovirus 71 epitopes elicit protective neutralizing antibodies in mice through divergent mechanisms.  

PubMed

Enterovirus 71 (EV71) is a major causative agent of hand, food, and mouth disease, which frequently occurs in young children. Since there are 11 subgenotypes (A, B1 to B5, and C1 to C5) within EV71, an EV71 vaccine capable of protecting against all of these subgenotypes is desirable. We report here the vaccine potential and protective mechanism of two chimeric virus-like particles (VLPs) presenting conserved neutralizing epitopes of EV71. We show that fusions of hepatitis B core antigen (HBc) with the SP55 or SP70 epitope of EV71, designated HBcSP55 and HBcSP70, respectively, can be rapidly generated and self-assembled into VLPs with the epitopes displayed on the surface. Immunization with the chimeric VLPs induced carrier- and epitope-specific antibody responses in mice. Anti-HBcSP55 and anti-HBcSP70 sera, but not anti-HBc sera, were able to neutralize in vitro multiple genotypes and strains of EV71. Importantly, passive immunization with anti-HBcSP55 or anti-HBcSP70 sera protected neonatal mice against lethal EV71 infections. Interestingly, anti-HBcSP70 sera could inhibit EV71 attachment to susceptible cells, whereas anti-HBcSP55 sera could not. However, both antisera were able to neutralize EV71 infection in vitro at the postattachment stage. The divergent mechanism of neutralization and protection conferred by anti-SP70 and anti-SP55 sera is in part attributed to their respective ability to bind authentic viral particles. Collectively, our study not only demonstrates that chimeric VLPs displaying the SP55 and SP70 epitopes are promising candidates for a broad-spectrum EV71 vaccine but also reveals distinct mechanisms of neutralization by the SP55- and SP70-targeted antibodies. PMID:24131712

Ye, Xiaohua; Ku, Zhiqiang; Liu, Qingwei; Wang, Xiaoli; Shi, Jinping; Zhang, Yunfang; Kong, Liangliang; Cong, Yao; Huang, Zhong

2014-01-01

197

1 5 O C T O B E R Human Papillomavirus  

E-print Network

1 5 O C T O B E R Human Papillomavirus Vaccination Programs and Human Immunodeficiency Virus the potential impact of human papillomavirus (HPV) vaccina- tion programs in developing countries; HPV, polygamy, cultural and economic factors), but they did not as- sess the potential effect of human im

Blower, Sally

198

Polyclonal human T lymphocyte activation results in the secondary functional activation of the human B lymphocyte.  

PubMed Central

The polyclonal T lymphocyte activators Con A and PHA were demonstrated to induce secretion of IgM and IgG as well as IgA antibodies in cultures of human peripheral blood lymphocytes. A similar response was seen in one-way mixed lymphocyte cultures and the kinetics, dose-response characteristics and optimal culture conditions are presented. The presence of functional T lymphocytes is a prerequisite for in vitro B lymphocyte activation in response to T lymphocyte mitogens. A narrow dose-response profile is a characteristic for both Con A and PHA and polyclonal B cell activation occurred at what have been previously regarded as suboptimal concentrations of these agents. The use of higher doses of these activators failed to generate immunoglobulin-secreting cells despite the presence of early and optimal levels of DNA synthesis in the cultures. PMID:6454516

Bird, A G; Hammarstrom, L; Smith, C I; Britton, S

1981-01-01

199

High-affinity interaction of hnRNP A1 with conserved RNA structural elements is required for translation and replication of enterovirus 71  

PubMed Central

Human Enterovirus 71 (EV71) is an emerging pathogen of infectious disease and a serious threat to public health. Currently, there are no antivirals or vaccines to slow down or prevent EV71 infections, thus underscoring the urgency to better understand mechanisms of host-enterovirus interactions. EV71 uses a type I internal ribosome entry site (IRES) to recruit the 40S ribosomal subunit via a pathway that requires the cytoplasmic localization of hnRNP A1, which acts as an IRES trans-activating factor. The mechanism of how hnRNP A1 trans activates EV71 RNA translation is unknown, however. Here, we report that the UP1 domain of hnRNP A1 interacts specifically with stem loop II (SLII) of the IRES, via a thermodynamically well-defined biphasic transition that involves conserved bulge 5?-AYAGY-3? and hairpin 5?-RY(U/A)CCA-3? loops. Calorimetric titrations of wild-type and mutant SLII constructs reveal these structural elements are essential to form a high-affinity UP1-SLII complex. Mutations that alter the bulge and hairpin primary or secondary structures abrogate the biphasic transition and destabilize the complex. Notably, mutations within the bulge that destabilize the complex correlate with a large reduction in IRES-dependent translational activity and impair EV71 replication. Taken together, this study shows that a conserved SLII structure is necessary to form a functional hnRNP A1-IRES complex, suggesting that small molecules that target this stem loop may have novel antiviral properties. PMID:23727900

Levengood, Jeffrey D.; Tolbert, Michele; Li, Mei-Ling; Tolbert, Blanton S.

2013-01-01

200

Gene expression analysis of primary normal human hepatocytes infected with human hepatitis B virus  

PubMed Central

AIM: To find the relationship between hepatitis B virus (HBV) and hepatocytes during the initial state of infection by cDNA microarray. METHODS: Primary normal human hepatocytes (PNHHs) were isolated and infected with HBV. From the PNHHs, RNA was isolated and inverted into complement DNA (cDNA) with Cy3- or Cy5- labeled dUTP for microarray analysis. The labeled cDNA was hybridized with microarray chip, including 4224 cDNAs. From the image of the microarray, expression profiles were produced and some of them were confirmed by RT-PCR, immunoblot analysis, and NF-?B luciferase reporter assay. RESULTS: From the cDNA microarray, we obtained 98 differentially regulated genes. Of the 98 genes, 53 were up regulated and 45 down regulated. Interestingly, in the up regulated genes, we found the TNF signaling pathway-related genes: LT-?, TRAF2, and NIK. By using RT-PCR, we confirmed the up-regulation of these genes in HepG2, Huh7, and Chang liver cells, which were transfected with pHBV1.2, a plasmid encoding all HBV messages. Moreover, these three genes participated in HBV-mediated NF-?B activation. CONCLUSION: During the initial state of HBV infection, hepatocytes facilitate the activation of NF-?B through up regulation of LT-?, TRAF2, and NIK. PMID:16937494

Ryu, Hyun Mi; Park, Sung Gyoo; Yea, Sung Su; Jang, Won Hee; Yang, Young-Il; Jung, Guhung

2006-01-01

201

Human monoclonal antibodies specific to hepatitis B virus generated in a human/mouse radiation chimera: the Trimera system.  

PubMed Central

An approach to develop fully human monoclonal antibodies in a human/mouse radiation chimera, the Trimera system, is described. In this system, functional human lymphocytes are engrafted in normal strains of mice which are rendered immuno-incompetent by lethal total body irradiation followed by radioprotection with severe combined immunodeficient (SCID) mouse bone marrow. Following transplantation, human lymphocytes colonize murine lymphatic organs and secrete human immunoglobulins. We have established this system as a tool to develop fully human monoclonal antibodies, and applied it for the generation of monoclonal antibodies specific for hepatitis B virus surface antigen. A strong memory response to hepatitis B surface antigen was elicited in Trimera engrafted with lymphocytes from human donors positive for antibodies to hepatitis B surface antigen. The human specific antibody fraction in the Trimera was 10(2)-10(3)-fold higher as compared with that found in the donors. Spleens were harvested from Trimera mice showing high specific-antibody titres and cells were fused to a human-mouse heteromyeloma fusion partner. Several stable hybridoma clones were isolated and characterized. These hybridomas produce high-affinity, IgG, anti-hepatitis B surface antigen antibodies demonstrating the potential of the Trimera system for generating fully human monoclonal antibodies. The biological function and the neutralizing activity of these antibodies are currently being tested. Images Figure 1 Figure 2 Figure 4 PMID:9616363

Eren, R; Lubin, I; Terkieltaub, D; Ben-Moshe, O; Zauberman, A; Uhlmann, R; Tzahor, T; Moss, S; Ilan, E; Shouval, D; Galun, E; Daudi, N; Marcus, H; Reisner, Y; Dagan, S

1998-01-01

202

Human regulatory B cells combine phenotypic and genetic hallmarks with a distinct differentiation fate.  

PubMed

Regulatory B cells (B-reg) produce IL-10 and suppress inflammation in both mice and humans, but limited data on the phenotype and function of these cells have precluded detailed assessment of their contribution to host immunity. In this article, we report that human B-reg cannot be defined based on a phenotype composed of conventional B cell markers, and that IL-10 production can be elicited in both the CD27(+) memory population and naive B cell subset after only a brief stimulation in vitro. We therefore sought to obtain a better definition of IL-10-producing human B-regs using a multiparameter analysis of B cell phenotype, function, and gene expression profile. Exposure to CpG and anti-Ig are the most potent stimuli for IL-10 secretion in human B cells, but microarray analysis revealed that human B cells cotreated with these reagents resulted in only ?0.7% of genes being differentially expressed between IL-10(+) and IL-10(-) cells. Instead, connectivity map analysis revealed that IL-10-secreting B cells are those undergoing specific differentiation toward a germinal center fate, and we identified a CD11c(+) B cell subset that was not capable of producing IL-10 even under optimal conditions. Our findings will assist in the identification of a broader range of human pro-B-reg populations that may represent novel targets for immunotherapy. PMID:25080484

Lin, Wenyu; Cerny, Daniela; Chua, Edmond; Duan, Kaibo; Yi, June Tai Jing; Shadan, Nurhidaya Binte; Lum, Josephine; Maho-Vaillant, Maud; Zolezzi, Francesca; Wong, Siew Cheng; Larbi, Anis; Fink, Katja; Musette, Philippe; Poidinger, Michael; Calbo, Sbastien

2014-09-01

203

Sporadic isolation of sabin-like polioviruses and high-level detection of non-polio enteroviruses during sewage surveillance in seven Italian cities, after several years of inactivated poliovirus vaccination.  

PubMed

Sewage surveillance in seven Italian cities between 2005 and 2008, after the introduction of inactivated poliovirus vaccination (IPV) in 2002, showed rare polioviruses, none that were wild-type or circulating vaccine-derived poliovirus (cVDPV), and many other enteroviruses among 1,392 samples analyzed. Two of five polioviruses (PV) detected were Sabin-like PV2 and three PV3, based on enzyme-linked immunosorbent assay (ELISA) and PCR results. Neurovirulence-related mutations were found in the 5'noncoding region (5'NCR) of all strains and, for a PV2, also in VP1 region 143 (Ile>Thr). Intertypic recombination in the 3D region was detected in a second PV2 (Sabin 2/Sabin 1) and a PV3 (Sabin 3/Sabin 2). The low mutation rate in VP1 for all PVs suggests limited interhuman virus passages, consistent with efficient polio immunization in Italy. Nonetheless, these findings highlight the risk of wild or Sabin poliovirus reintroduction from abroad. Non-polio enteroviruses (NPEVs) were detected, 448 of which were coxsackievirus B (CVB) and 294 of which were echoviruses (Echo). Fifty-six NPEVs failing serological typing were characterized by sequencing the VP1 region (nucleotides [nt] 2628 to 2976). A total of 448 CVB and 294 Echo strains were identified; among those strains, CVB2, CVB5, and Echo 11 predominated. Environmental CVB5 and CVB2 strains from this study showed high sequence identity with GenBank global strains. The high similarity between environmental NPEVs and clinical strains from the same areas of Italy and the same periods indicates that environmental strains reflect the viruses circulating in the population and highlights the potential risk of inefficient wastewater treatments. This study confirmed that sewage surveillance can be more sensitive than acute flaccid paralysis (AFP) surveillance in monitoring silent poliovirus circulation in the population as well as the suitability of molecular approaches to enterovirus typing. PMID:24814793

Battistone, A; Buttinelli, G; Fiore, S; Amato, C; Bonomo, P; Patti, A M; Vulcano, A; Barbi, M; Binda, S; Pellegrinelli, L; Tanzi, M L; Affanni, P; Castiglia, P; Germinario, C; Mercurio, P; Cicala, A; Triassi, M; Pennino, F; Fiore, L

2014-08-01

204

Approaches to design non-covalent inhibitors for human granzyme B (hGrB).  

PubMed

A structure-based design campaign for non-covalent small molecule inhibitors of human granzyme B was carried out by means of a virtual screening strategy employing three constraints and probe site-mapping with FTMAP to identify ligand "hot spots". In addition, new scaffolds of diverse structures were subsequently explored with ROCS shape-based superposition methods, following by Glide SP docking, induced fit docking and analysis of QikProp molecular properties. Novel classes of moderately active small molecule blockers (?25 ?M IC50 values) from commercially available libraries were identified, and three novel scaffolds have been synthesized by multi-step procedures. Furthermore, we provide an example of a comprehensive structure-based drug discovery approach to non-covalent inhibitors that relies on the X-ray structure of a covalently bound ligand and suggest that the design path may be compromised by alternative and unknown binding poses. PMID:25277547

Kim, Mi-Sun; Buisson, Lauriane A; Heathcote, Dean A; Hu, Haipeng; Braddock, D Christopher; Barrett, Anthony G M; Ashton-Rickardt, Philip G; Snyder, James P

2014-10-21

205

The identification of a phospholipase B precursor in human neutrophils.  

PubMed

A phospholipase B (PLB) precursor was purified from normal human granulocytes using Sephadex G-75, Mono-S cation-exchange and hydroxyapatite columns. The molecular mass of the protein was estimated to be approximately 130 kDa by gel filtration and 22 and 42 kDa by SDS/PAGE. Tryptic peptide and sequence analyses by MALDI-TOF and tandem mass spectrometry (MS/MS) identified the protein as a FLJ22662 (Homo sapiens) gene product, a homologue of the amoeba Dictyostelium discoideum PLB. The native protein needed modifications to acquire deacylation activity against phospholipids including phosphatidylcholine, phosphatidylinositol, phosphatidylethanolamine and lysophospholipids. Enzyme activity was associated with fragments derived from the 42 kDa fragment. The enzyme revealed a PLB nature by removing fatty acids from both the sn-1 and sn-2 positions of phospholipids. The enzyme is active at a broad pH range with an optimum of 7.4. Immunoblotting of neutrophil postnuclear supernatant using antibodies against the 42 kDa fragment detected a band at a molecular mass of 42 kDa, indicating a neutrophil origin of the novel PLB precursor. The existence of the PLB precursor in neutrophils and its enzymatic activity against phospholipids suggest a role in the defence against invading microorganisms and in the generation of lipid mediators of inflammation. PMID:19019078

Xu, Shengyuan; Zhao, Linshu; Larsson, Anders; Venge, Per

2009-01-01

206

Metabolism of leukotriene B4 by activated human polymorphonuclear granulocytes.  

PubMed Central

Human polymorphonuclear granulocytes (PMNs) synthesize leukotriene B4 (LTB4) as a response of cell activation. Inactivation of the potent inflammatory mediator proceeds via omega-oxidation, resulting in the formation of 20-hydroxy- and 20-carboxy-LTB4. The main metabolite after stimulation with the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (FMLP) is 20-carboxy-LTB4, and after stimulation with the calcium ionophore A23187 is 20-hydroxy-LTB4. Differences in the LTB4 inactivation pathway were also observed when the catabolism of exogenously added LTB4 was analysed. In contrast to resting cells or cells preactivated with FMLP, prestimulation with the ionophore or with phorbol esters resulted in the inhibition of 20-carboxy-LTB4-generation. This decrease correlated with the reduction in specific [3H] LTB4-receptor expression. Studies with the non-penetrating diazonium salt of sulphanilic acid, which is known to interact with ectoenzymes, revealed that LTB4 is metabolized via receptor-mediated uptake. Our data suggest that the reduction in the amount of LTB4-receptor sites inhibits the conversion of 20-OH-LTB4 into 20-COOH-LTB4. PMID:2842254

Brom, J; Schonfeld, W; Konig, W

1988-01-01

207

Feto-placentary pathology in human parvovirus B19 infection.  

PubMed

In view of the scarce references concerning the histological data in congenital parvovirus human B19 infection, we intend to provide a description of the pathological features observed in six autopsies. The virus was detected by DNA hybridization (ISH-DBH), PCR and electronmicroscopy (EM) in paraffin-embedded feto-placentary tissues. These cases constitute a subset from 86 Non Immunologic Hydrops Fetalis (NIHF) cases, in which a systemic complex of inflammatory/degenerative lesions of unknown etiology was visualized by optical microscopy. In one case a syphilitic process was detected, typefying a double infection. All fetuses showed a similar pathology--hydrops, hepato-splenomegaly, lung hypoplasia and erythroblastemia, the specific histological feature being the presence of intranuclear inclusions in the erythroid progenitors, in the erythropoietic visceral tissue and in blood marrow. Complex cardiopathy allied to abnormal lung lobulation and polisplenia were observed once; in 2 cases endocardial fibroelastosis was diagnosed. The pulmonary lesions were represented by dysmaturity allied to interstitial mononuclear infiltration. The hepatic consisted of cholestasis, portal fibrosis, canalicular proliferation, hemossiderosis, focal necroses and giant cell transformation. The central nervous system lesions were predominantly anoxic although the autolysis impaired a correct diagnosis. PMID:9830727

Garcia, A G; Pegado, C S; Cubel, R de C; Fonseca, M E; Sloboda, I; Nascimento, J P

1998-01-01

208

Role of hepatitis B virus DNA integration in human hepatocarcinogenesis  

PubMed Central

Liver cancer ranks sixth in cancer incidence, and is the third leading cause of cancer-related deaths worldwide. Hepatocellular carcinoma (HCC) is the most common type of liver cancer, which arises from hepatocytes and accounts for approximately 70%-85% of cases. Hepatitis B virus (HBV) frequently causes liver inflammation, hepatic damage and subsequent cirrhosis. Integrated viral DNA is found in 85%-90% of HBV-related HCCs. Its presence in tumors from non-cirrhotic livers of children or young adults further supports the role of viral DNA integration in hepatocarcinogenesis. Integration of subgenomic HBV DNA fragments into different locations within the host DNA is a significant feature of chronic HBV infection. Integration has two potential consequences: (1) the host genome becomes altered (cis effect); and (2) the HBV genome becomes altered (trans effect). The cis effect includes insertional mutagenesis, which can potentially disrupt host gene function or alter host gene regulation. Tumor progression is frequently associated with rearrangement and partial gain or loss of both viral and host sequences. However, the role of integrated HBV DNA in hepatocarcinogenesis remains controversial. Modern technology has provided a new paradigm to further our understanding of disease mechanisms. This review summarizes the role of HBV DNA integration in human carcinogenesis. PMID:24876744

Hai, Hoang; Tamori, Akihiro; Kawada, Norifumi

2014-01-01

209

Role of hepatitis B virus DNA integration in human hepatocarcinogenesis.  

PubMed

Liver cancer ranks sixth in cancer incidence, and is the third leading cause of cancer-related deaths worldwide. Hepatocellular carcinoma (HCC) is the most common type of liver cancer, which arises from hepatocytes and accounts for approximately 70%-85% of cases. Hepatitis B virus (HBV) frequently causes liver inflammation, hepatic damage and subsequent cirrhosis. Integrated viral DNA is found in 85%-90% of HBV-related HCCs. Its presence in tumors from non-cirrhotic livers of children or young adults further supports the role of viral DNA integration in hepatocarcinogenesis. Integration of subgenomic HBV DNA fragments into different locations within the host DNA is a significant feature of chronic HBV infection. Integration has two potential consequences: (1) the host genome becomes altered ("cis" effect); and (2) the HBV genome becomes altered ("trans" effect). The cis effect includes insertional mutagenesis, which can potentially disrupt host gene function or alter host gene regulation. Tumor progression is frequently associated with rearrangement and partial gain or loss of both viral and host sequences. However, the role of integrated HBV DNA in hepatocarcinogenesis remains controversial. Modern technology has provided a new paradigm to further our understanding of disease mechanisms. This review summarizes the role of HBV DNA integration in human carcinogenesis. PMID:24876744

Hai, Hoang; Tamori, Akihiro; Kawada, Norifumi

2014-05-28

210

Detection of enterovirus 68 in serum from pediatric patients with pneumonia and their clinical outcomes  

PubMed Central

Enterovirus 68 (EV68) infection occasionally manifests with fatal outcomes. However, detection of EV68 in serum and its clinical outcomes are yet to be determined. In this study, we retrospectively tested stored serum samples collected from pediatric pneumonia patients whose nasopharyngeal specimens were positive for EV68. Of total 28 nasopharyngeal sample-positive patients, EV68 was detected in serum samples among 12 (43%) patients aged between 1 and 4 years. Our results suggest that EV68 can cause viremia by which the virus may exhibit systemic manifestations. PMID:24209770

Imamura, Tadatsugu; Suzuki, Akira; Lupisan, Socorro; Kamigaki, Taro; Okamoto, Michiko; Roy, Chandra Nath; Olveda, Remigio; Oshitani, Hitoshi

2014-01-01

211

Spectromicroscopy of boron in human glioblastomas following administration of Na2B12H11SH B. Gilbert,1,  

E-print Network

Spectromicroscopy of boron in human glioblastomas following administration of Na2B12H11SH B glioblastoma tissue from two patients administered and one not administered with BSH. As well as weak signals number s : 87.59. e I. INTRODUCTION Glioblastoma multiforme is the most malignant form of glioma cancer

Gilbert, Pupa Gelsomina De Stasio

212

Identification and mapping of neutralizing epitopes of human parvovirus B19 by using human antibodies.  

PubMed Central

We identified and mapped the regions responsible for neutralization in the human parvovirus B19 structural protein by using region-specific human antibodies derived from seropositive blood donors. The region-specific antibodies were purified by using affinity columns coupled with synthetic peptides of the hydrophilic regions including the beta-turn structure deduced by the predicted secondary structure of VP2. Fifteen highly specific antibodies against the synthetic peptides were obtained. Ten of them were able to precipitate the radiolabeled virus. Six of them proved to be able to protect the colony-forming unit erythroid cells in human bone marrow cell cultures from injury by the virus. The sequences recognized by the six neutralizing antibodies were sites corresponding to amino acids 253 to 272, 309 to 330, 325 to 346, 359 to 382, 449 to 468, and 491 to 515 from the amino-terminal portion of VP2. These observations suggest that the neutralizing epitopes were distributed in the region from amino acid 253 in the amino-terminal portion of VP2 to the carboxyl terminus of VP2. PMID:1716693

Sato, H; Hirata, J; Kuroda, N; Shiraki, H; Maeda, Y; Okochi, K

1991-01-01

213

Posttranslational modification of I-kappa B alpha activates NF?B in human monocytes exposed to 56 Fe ions  

Microsoft Academic Search

The objective of this study was to investigate whether heavy ion (56Fe) radiation exposure activates one of the key transcriptional regulators, nuclear factor-kappa B (NF-3B), in normal human monocytes (Mono Mac 6 cells: MM6). The study revealed that the exposure of MM6 cells to 56Fe ions resulted in increased NF-3B DNA-binding activity. The activation was both dose- and time-dependent, with

Mohan Natarajan; N. Aravindan; M. L. Meltz; T. S. Herman

2002-01-01

214

Parvovirus B19 DNA Is Commonly Harboured in Human Skin  

Microsoft Academic Search

Background: Parvovirus B19 is the aetiological agent of erythema infectiosum. The presence of B19 DNA in lesional skin of other cutaneous manifestations has frequently been reported although there is disagreement on the role of the B19 virus in tissues. Objectives: To investigate the presence of B19 DNA (1) in skin lesions of patients with a described B19-related disease, (2) in

F. Bonvicini; M. La Placa; E. Manaresi; G. Gallinella; G. A. Gentilomi; M. Zerbini; M. Musiani

2010-01-01

215

1,25-dihydroxyvitamin D{sub 3} impairs NF-{kappa}B activation in human naive B cells  

SciTech Connect

Highlights: {yields} In naive B cells, VDR activation by calcitriol results in reduced NF-{kappa}B p105 and p50 protein expression. {yields} Ligating the VDR with calcitriol causes reduced nuclear translocation of NF-{kappa}B p65. {yields} Reduced nuclear amount of p65 after calcitriol incubation results in reduced binding of p65 on the p105 promoter. {yields} Thus, vitamin D receptor signaling may reduce or prevent activation of B cells and unwanted immune responses, e.g. in IgE dependent diseases such as allergic asthma. -- Abstract: 1{alpha},25-dihydroxyvitamin D{sub 3} (calcitriol), the bioactive metabolite of vitamin D, modulates the activation and inhibits IgE production of anti-CD40 and IL-4 stimulated human peripheral B cells. Engagement of CD40 results in NF-{kappa}B p50 activation, which is essential for the class switch to IgE. Herein, we investigated by which mechanism calcitriol modulates NF-{kappa}B mediated activation of human naive B cells. Naive B cells were predominantly targeted by calcitriol in comparison with memory B cells as shown by pronounced induction of the VDR target gene cyp24a1. Vitamin D receptor activation resulted in a strongly reduced p105/p50 protein and mRNA expression in human naive B cells. This effect is mediated by impaired nuclear translocation of p65 and consequently reduced binding of p65 to its binding site in the p105 promoter. Our data indicate that the vitamin D receptor reduces NF-{kappa}B activation by interference with NF-{kappa}B p65 and p105. Thus, the vitamin D receptor inhibits costimulatory signal transduction in naive B cells, namely by reducing CD40 signaling.

Geldmeyer-Hilt, Kerstin, E-mail: kerstin.hilt@charite.de [Allergie-Centrum-Charite, CCM, Klinik fuer Dermatologie und Allergologie, Charite - Universitaetsmedizin Berlin, Chariteplatz 1, 10117 Berlin (Germany)] [Allergie-Centrum-Charite, CCM, Klinik fuer Dermatologie und Allergologie, Charite - Universitaetsmedizin Berlin, Chariteplatz 1, 10117 Berlin (Germany); Heine, Guido, E-mail: guido.heine@charite.de [Allergie-Centrum-Charite, CCM, Klinik fuer Dermatologie und Allergologie, Charite - Universitaetsmedizin Berlin, Chariteplatz 1, 10117 Berlin (Germany) [Allergie-Centrum-Charite, CCM, Klinik fuer Dermatologie und Allergologie, Charite - Universitaetsmedizin Berlin, Chariteplatz 1, 10117 Berlin (Germany); Deutsches Rheuma-Forschungszentrum Berlin, Chariteplatz 1, 10117 Berlin (Germany); Hartmann, Bjoern, E-mail: bjoern.hartmann@charite.de [Allergie-Centrum-Charite, CCM, Klinik fuer Dermatologie und Allergologie, Charite - Universitaetsmedizin Berlin, Chariteplatz 1, 10117 Berlin (Germany)] [Allergie-Centrum-Charite, CCM, Klinik fuer Dermatologie und Allergologie, Charite - Universitaetsmedizin Berlin, Chariteplatz 1, 10117 Berlin (Germany); Baumgrass, Ria, E-mail: baumgrass@drfz.de [Deutsches Rheuma-Forschungszentrum Berlin, Chariteplatz 1, 10117 Berlin (Germany)] [Deutsches Rheuma-Forschungszentrum Berlin, Chariteplatz 1, 10117 Berlin (Germany); Radbruch, Andreas, E-mail: radbruch@drfz.de [Deutsches Rheuma-Forschungszentrum Berlin, Chariteplatz 1, 10117 Berlin (Germany)] [Deutsches Rheuma-Forschungszentrum Berlin, Chariteplatz 1, 10117 Berlin (Germany); Worm, Margitta, E-mail: margitta.worm@charite.de [Allergie-Centrum-Charite, CCM, Klinik fuer Dermatologie und Allergologie, Charite - Universitaetsmedizin Berlin, Chariteplatz 1, 10117 Berlin (Germany)] [Allergie-Centrum-Charite, CCM, Klinik fuer Dermatologie und Allergologie, Charite - Universitaetsmedizin Berlin, Chariteplatz 1, 10117 Berlin (Germany)

2011-04-22

216

Studies of lymphocyte reconstitution in a humanized mouse model reveal a requirement of T cells for human B cell maturation  

PubMed Central

The hematopoietic humanized mouse (hu-mouse) model is a powerful resource to study and manipulate the human immune system. However, a major and recurrent issue with this model has been the poor maturation of B cells that fail to progress beyond the transitional B cell stage. Interestingly, a similar problem has been reported in transplant patients that receive cord blood stem cells. In this study, we characterize the development of human B and T cells in the lymph nodes (LNs) and spleen of BALB/c-Rag2nullIl2r?null hu-mice. We find a dominant population of immature B cells in the blood and spleen early, followed by a population of human T cells, coincident with the detection of LNs. Notably, in older mice we observe a major population of mature B cells in LNs and in the spleens of mice with higher T cell frequencies. Moreover, we demonstrate that the T cells are necessary for B cell maturation, as introduction of autologous human T cells expedites the appearance of mature B cells, while in vivo depletion of T cells retards B cell maturation. The presence of the mature B cell population correlates with enhanced IgG and Ag-specific responses to both T-dependent and T-independent challenges, indicating their functionality. These findings enhance our understanding of human B cell development, provide increased details of the reconstitution dynamics of hu-mice, and validates the use of this animal model to study mechanisms and treatments for the similar delay of functional B cells associated with cord blood transplantations. PMID:23335750

Lang, Julie; Kelly, Margot; Freed, Brian M.; McCarter, Martin D.; Kedl, Ross M.; Torres, Raul M.; Pelanda, Roberta

2013-01-01

217

Structural basis for the recognition of human cytomegalovirus glycoprotein B by a neutralizing human antibody.  

PubMed

Human cytomegalovirus (HCMV) infections are life-threating to people with a compromised or immature immune system. Upon adhesion, fusion of the virus envelope with the host cell is initiated. In this step, the viral glycoprotein gB is considered to represent the major fusogen. Here, we present for the first time structural data on the binding of an anti-herpes virus antibody and describe the atomic interactions between the antigenic domain Dom-II of HCMV gB and the Fab fragment of the human antibody SM5-1. The crystal structure shows that SM5-1 binds Dom-II almost exclusively via only two CDRs, namely light chain CDR L1 and a 22-residue-long heavy chain CDR H3. Two contiguous segments of Dom-II are targeted by SM5-1, and the combining site includes a hydrophobic pocket on the Dom-II surface that is only partially filled by CDR H3 residues. SM5-1 belongs to a series of sequence-homologous anti-HCMV gB monoclonal antibodies that were isolated from the same donor at a single time point and that represent different maturation states. Analysis of amino acid substitutions in these antibodies in combination with molecular dynamics simulations show that key contributors to the picomolar affinity of SM5-1 do not directly interact with the antigen but significantly reduce the flexibility of CDR H3 in the bound and unbound state of SM5-1 through intramolecular side chain interactions. Thus, these residues most likely alleviate unfavorable binding entropies associated with extra-long CDR H3s, and this might represent a common strategy during antibody maturation. Models of entire HCMV gB in different conformational states hint that SM5-1 neutralizes HCMV either by blocking the pre- to postfusion transition of gB or by precluding the interaction with additional effectors such as the gH/gL complex. PMID:25299639

Spindler, Nadja; Diestel, Uschi; Stump, Joachim D; Wiegers, Anna-Katharina; Winkler, Thomas H; Sticht, Heinrich; Mach, Michael; Muller, Yves A

2014-10-01

218

Structural Basis for the Recognition of Human Cytomegalovirus Glycoprotein B by a Neutralizing Human Antibody  

PubMed Central

Human cytomegalovirus (HCMV) infections are life-threating to people with a compromised or immature immune system. Upon adhesion, fusion of the virus envelope with the host cell is initiated. In this step, the viral glycoprotein gB is considered to represent the major fusogen. Here, we present for the first time structural data on the binding of an anti-herpes virus antibody and describe the atomic interactions between the antigenic domain Dom-II of HCMV gB and the Fab fragment of the human antibody SM5-1. The crystal structure shows that SM5-1 binds Dom-II almost exclusively via only two CDRs, namely light chain CDR L1 and a 22-residue-long heavy chain CDR H3. Two contiguous segments of Dom-II are targeted by SM5-1, and the combining site includes a hydrophobic pocket on the Dom-II surface that is only partially filled by CDR H3 residues. SM5-1 belongs to a series of sequence-homologous anti-HCMV gB monoclonal antibodies that were isolated from the same donor at a single time point and that represent different maturation states. Analysis of amino acid substitutions in these antibodies in combination with molecular dynamics simulations show that key contributors to the picomolar affinity of SM5-1 do not directly interact with the antigen but significantly reduce the flexibility of CDR H3 in the bound and unbound state of SM5-1 through intramolecular side chain interactions. Thus, these residues most likely alleviate unfavorable binding entropies associated with extra-long CDR H3s, and this might represent a common strategy during antibody maturation. Models of entire HCMV gB in different conformational states hint that SM5-1 neutralizes HCMV either by blocking the pre- to postfusion transition of gB or by precluding the interaction with additional effectors such as the gH/gL complex. PMID:25299639

Stump, Joachim D.; Wiegers, Anna-Katharina; Winkler, Thomas H.; Sticht, Heinrich; Mach, Michael; Muller, Yves A.

2014-01-01

219

Human immunodeficiency virus-1 Tat activates NF-?B via physical interaction with I?B-? and p65  

PubMed Central

Nuclear factor (NF)-?B is a master regulator of pro-inflammatory genes and is upregulated in human immunodeficiency virus 1 (HIV-1) infection. Mechanisms underlying the NF-?B deregulation by HIV-1 are relevant for immune dysfunction in AIDS. We report that in single round HIV-1 infection, or single-pulse PMA stimulation, the HIV-1 Tat transactivator activated NF-?B by hijacking the inhibitor I?B-? and by preventing the repressor binding to the NF-?B complex. Moreover, Tat associated with the p65 subunit of NF-?B and increased the p65 DNA-binding affinity and transcriptional activity. The arginine- and cysteine-rich domains of Tat were required for I?B-? and p65 association, respectively, and for sustaining the NF-?B activity. Among an array of NF-?B-responsive genes, Tat mostly activated the MIP-1? expression in a p65-dependent manner, and bound to the MIP-1? NF-?B enhancer thus promoting the recruitment of p65 with displacement of I?B-?; similar findings were obtained for the NF-?B-responsive genes CSF3, LTA, NFKBIA and TLR2. Our results support a novel mechanism of NF-?B activation via physical interaction of Tat with I?B-? and p65, and may contribute to further insights into the deregulation of the inflammatory response by HIV-1. PMID:22187158

Fiume, Giuseppe; Vecchio, Eleonora; De Laurentiis, Annamaria; Trimboli, Francesca; Palmieri, Camillo; Pisano, Antonio; Falcone, Cristina; Pontoriero, Marilena; Rossi, Annalisa; Scialdone, Annarita; Fasanella Masci, Francesca; Scala, Giuseppe; Quinto, Ileana

2012-01-01

220

Expression and localization of a UT-B urea transporter in the human bladder.  

PubMed

Facilitative UT-B urea transporters have been shown to play an important role in the urinary concentrating mechanism. Recent studies have now suggested a link between UT-B allelic variation and human bladder cancer risk. UT-B1 protein has been previously identified in the bladder of various mammalian species, but not yet in humans. The aim of the present study was to investigate whether any UT-B protein was present in the human bladder. First, RT-PCR results confirmed that UT-B1 was strongly expressed at the RNA level in the human bladder, whereas UT-B2 was only weakly present. Initial Western blot analysis confirmed that a novel UT-B COOH-terminal antibody detected human UT-B proteins. Importantly, this antibody detected a specific 40- to 45-kDa UT-B signal in human bladder protein. Using a peptide-N-glycosidase F enzyme, this bladder UT-B signal was deglycosylated to a core 30-kDa protein, which is smaller than the predicted size for UT-B1 but similar to many proteins reported to be UT-B1. Finally, immunolocalization experiments confirmed that UT-B protein was strongly expressed throughout all urothelium layers except for the apical membrane of the outermost umbrella cells. In conclusion, these data confirm the presence of UT-B protein within the human bladder. Further studies are now required to determine the precise nature, regulation, and physiological role of this UT-B. PMID:25209859

Walpole, C; Farrell, A; McGrane, A; Stewart, G S

2014-11-01

221

Identification of DNA of human origin based on amplification of human-specific mitochondrial cytochrome b region  

Microsoft Academic Search

Species-specific differences in a non-polymorphic region of the mitochondrial cytochrome b gene appear to be large enough to allow human-specific amplification of forensic DNA samples. We therefore developed a PCR-based method using newly designed primers to amplify a 157-bp portion of the human mitochondrial cytochrome b gene. The forward and reverse primers were designed to hybridize to regions of the

Hirokazu Matsuda; Yasuhisa Seo; Eiji Kakizaki; Shuji Kozawa; Eri Muraoka; Nobuhiro Yukawa

2005-01-01

222

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

223

Regulation of VH Replacement by B Cell Receptor (BCR)-mediated Signaling in Human Immature B Cells  

PubMed Central

VH replacement provides a unique RAG-mediated recombination mechanism to edit non-functional IgH genes or IgH genes encoding self reactive B cell receptors (BCRs) and contributes to the diversification of antibody repertoire in mouse and human. Currently, it is not clear how VH replacement is regulated during early B lineage cell development. Here we show that crosslinking BCRs induces VH replacement in human EU12 ?HC+ cells and in the newly emigrated immature B cells purified from peripheral blood of healthy donors or tonsillar samples. BCR signaling-induced VH replacement is dependent on the activation of Syk and Src kinases; but is inhibited by CD19 co-stimulation, presumably through activation of the PI3 kinase pathway. These results show for the first time that VH replacement is regulated by BCR-mediated signaling in human immature B cells, which can be modulated by physiological and pharmacological treatments. PMID:23630348

Liu, Jing; Lange, Miles D.; Hong, Sang Yong; Xie, Wanqin; Xu, Kerui; Huang, Lin; Yu, Yangsheng; Ehrhardt, Gotz R. A.; Zemlin, Michael; Burrows, Peter D.; Su, Kaihong; Carter, Robert H.; Zhang, Zhixin

2013-01-01

224

SLEEP-DEPENDENT MOTOR MEMORY PLASTICITY IN THE HUMAN M. P. WALKER,a,b  

E-print Network

SLEEP-DEPENDENT MOTOR MEMORY PLASTICITY IN THE HUMAN BRAIN M. P. WALKER,a,b * R. STICKGOLD,b D of sleep- dependent learning in both humans and animals (Walker and Stickgold, 2004), already complemented reviews see Walker, in press). Demonstrations of over- night, sleep-dependent learning have now been

Walker, Matthew P.

225

Early B lymphocyte development: Similarities and differences in human and mouse  

PubMed Central

B lymphocytes differentiate from hematopoietic stem cells through a series of distinct stages. Early B cell development proceeds in bone marrow until immature B cells migrate out to secondary lymphoid tissues, such as a spleen and lymph nodes, after completion of immunoglobulin heavy and light chain rearrangement. Although the information about the regulation by numerous factors, including signaling molecules, transcription factors, epigenetic changes and the microenvironment, could provide the clinical application, our knowledge on human B lymphopoiesis is limited. However, with great methodological advances, significant progress for understanding B lymphopoiesis both in human and mouse has been made. In this review, we summarize the experimental models for studies about human adult B lymphopoiesis, and the role of microenvironment and signaling molecules, such as cytokines, transforming growth factor-? superfamily, Wnt family and Notch family, with point-by-point comparison between human and mouse.

Ichii, Michiko; Oritani, Kenji; Kanakura, Yuzuru

2014-01-01

226

Characterization of an Enterovirus species E isolated from naturally infected bovine in China.  

PubMed

Bovine enteroviruses, which belong to the Picornaviridae family, can cause clinical symptoms in cattle and are excreted in feces. In this study, a cytolytic virus was isolated from Madin-Darby bovine kidney (MDBK) cells from fecal samples of bovine with severe diarrhea and hemorrhagic intestinal mucosa that had been originally diagnosed with bovine viral diarrhea (BVD) by a bovine viral diarrhea virus Ag point-of-care test (IDEXX, American). Random priming PCR was used to amplify underlying viral sequences and identify the isolated virus. Phylogenetic analysis indicated that the isolated virus closely matches the EV-E2 species, which is different from other Chinese strains previously isolated. The newly identified virus was named HLJ-3531/2013. We infected the sulking mice with the isolated virus. Reverse-transcription PCR, hematoxylin and eosin (HE) staining, serum neutralization (SN) test, and virus isolation from various tissues revealed that HLJ-3531/2013 can infect the intestine, liver, and lung of suckling mice. The present work is the first to report the reproduction of clinical symptoms by an isolated virus in an experimental infection model of animals and lays a solid foundation for the development of the pathogenesis of bovine enteroviruses. PMID:25102330

Zhang, Haili; Liu, Hongtao; Bao, Jun; Guo, Yongli; Peng, Tongquan; Zhou, Pingping; Zhang, Wenlong; Ma, Bo; Wang, Junwei; Gao, Mingchun

2014-10-13

227

Molecular surveillance of non-polio enterovirus infections in patients with acute gastroenteritis in Western India: 2004-2009.  

PubMed

Acute gastroenteritis is a major cause of childhood morbidity and mortality worldwide. Rotavirus (RV) and Norovirus (NoV) are the leading cause of the disease. Despite the use of improved diagnostic methods a significant proportion of gastroenteritis cases remained undiagnosed. Though nonpolio enteroviruses (NPEVs) have been reported frequently in children with acute gastroenteritis, their etiologic role has not been established. To investigate the epidemiology of NPEVs in gastroenteritis cases which remained negative for leading causative agents, 955 RV and NoV negative stool specimens from children hospitalized for acute gastroenteritis were included in the study. A case control study was conducted which includes stool specimens from 450 children with gastroenteritis and 162 asymptomatic control subjects to determine the association of NPEVs with the disease. NPEV detection and typing was carried out by RT-PCR and sequencing. Presence of RV, NoV, Adenovirus, and Astrovirus was confirmed by ELISA or PCR/RT-PCR. Overall 14% NPEV prevalence was noted. The percentage of children with NPEV infection differed significantly between gastroenteritis and non-gastroenteritis patients (13.7% vs. 4.9%). NPEV was more prevalent among patients with gastroenteritis of undetectable etiology as compared to those detected positive for other viruses (17.9% vs. 7%) (P?B species (56.5%) followed by EV-C (16.7%), EV-A (13.8%) species and mixed NPEV infections (13%). These data support the association of NPEVs with acute gastroenteritis and highlights the clinical and epidemiological features of NPEV infections in patients with acute gastroenteritis from western India. J. Med. Virol. 87: 154-161, 2015. 2014 Wiley Periodicals, Inc. PMID:24903844

Patil, Pooja R; Chitambar, Shobha D; Gopalkrishna, V

2015-01-01

228

Molecular Surveillance of Enterovirus and Norwalk-Like Virus in Oysters Relocated to a Municipal-Sewage-Impacted Gulf Estuary  

Microsoft Academic Search

An 18-month survey was conducted to examine the prevalence of enteric viruses and their relationship to indicators in environmentally polluted shellfish. Groups of oysters, one group per 4 weeks, were relocated to a coastal water area in the Gulf of Mexico that is impacted by municipal sewage and were analyzed for enteroviruses, Norwalk-like viruses (NLV), and indicator microorganisms (fecal coliform,

Y. Carol Shieh; Ralph S. Baric; Jacquelina W. Woods; Kevin R. Calci

2003-01-01

229

The Seroprevalence and Seroincidence of Enterovirus71 Infection in Infants and Children in Ho Chi Minh City, Viet Nam  

Microsoft Academic Search

Enterovirus 71 (EV71)-associated hand, foot and mouth disease has emerged as a serious public health problem in South East Asia over the last decade. To better understand the prevalence of EV71 infection, we determined EV71 seroprevalence and seroincidence amongst healthy infants and children in Ho Chi Minh City, Viet Nam. In a cohort of 200 newborns, 55% of cord blood

Chau Bich Nguyen Tran; Hieu Trong Nguyen; Ha Thanh Thi Phan; Ngoc Van Tran; Bridget Wills; Jeremy Farrar; Joseph D. Santangelo; Cameron P. Simmons

2011-01-01

230

A Human Leukemia Cell with Both B and T Cell Surface Receptors  

PubMed Central

Bone-marrow-derived (B) and thymus-derived (T) lymphocytes can be distinguished by the presence of a number of receptors and differentiation antigens. The presence of these markers has facilitated the identification and characterization of the mononuclear cells in a number of animal and human lymphoid malignancies. We describe here the immunological properties of human leukemia cells that are highly unusual, since they simultaneously bear the receptor for sheep erythrocytes characteristic of human T lymphocytes and the receptor for antigen-antibody-complement complexes characteristic of human B lymphocytes. A small number (about 2%) of normal human lymphocytes bearing both of these receptors was also identified. Images PMID:4274561

Shevach, E.; Edelson, R.; Frank, M.; Lutzner, M.; Green, I.

1974-01-01

231

Mixed-mode fracture of human cortical bone Elizabeth A. Zimmermann a,b  

E-print Network

Mixed-mode fracture of human cortical bone Elizabeth A. Zimmermann a,b , Maximilien E. Launey Available online 1 July 2009 Keywords: Human cortical bone Mixed-mode fracture Fracture toughness Fracture of most fracture mechanics studies of human cortical bone, bones in vivo are invariably loaded

Ritchie, Robert

232

A Taxonomy of Distributed Human Computation Alexander J. Quinn, Benjamin B. Bederson  

E-print Network

A Taxonomy of Distributed Human Computation Alexander J. Quinn, Benjamin B. Bederson University of Maryland Department of Computer Science Human-Computer Interaction Lab College Park, MD 20742 USA {aq, bederson}@cs.umd.edu ABSTRACT Distributed Human Computation (DHC) holds great promise for using computers

Golbeck, Jennifer

233

H222c Introduction to Energy & Environment: Humans & Nature P. B. Rhines  

E-print Network

and The Population Explosion where it is called `PAT' as an aid to memory. McKibben's book Eaarth deals with humans. The human body, viewed as an engine that converts chemical energy from food into glucoseH222c Introduction to Energy & Environment: Humans & Nature P. B. Rhines spring 2012 Lecture 2 I

234

Phase behavior of mixtures of human lens proteins Gamma D and Beta B1  

E-print Network

mgmL (which is approximately the total protein concentration in a human eye lens) (14­17). Significant%) crystallins in human eye lens (20), studies of their mix- tures are of particular importance. In relationPhase behavior of mixtures of human lens proteins Gamma D and Beta B1 Ying Wanga , Aleksey Lomakina

Benedek, George B.

235

Tracking human antigen-specific memory B cells: a sensitive and generalized ELISPOT system  

Microsoft Academic Search

In the interest of better understanding the role of human memory B cells in protection against disease, we developed an assay to quantitate antigen-specific memory B cells in human blood. This assay utilizes a 6-day polyclonal stimulation of PBMC followed by an antigen-specific ELISPOT for the detection of memory B cells that have differentiated into antibody secreting cells (ASC) in

Shane Crotty; Rachael D. Aubert; John Glidewell; Rafi Ahmed

2004-01-01

236

Human Genome Diversity Project. Summary of planning workshop 3(B): Ethical and human-rights implications  

SciTech Connect

The third planning workshop of the Human Genome Diversity Project was held on the campus of the US National Institutes of Health in Bethesda, Maryland, from February 16 through February 18, 1993. The second day of the workshop was devoted to an exploration of the ethical and human-rights implications of the Project. This open meeting centered on three roundtables, involving 12 invited participants, and the resulting discussions among all those present. Attendees and their affiliations are listed in the attached Appendix A. The discussion was guided by a schedule and list of possible issues, distributed to all present and attached as Appendix B. This is a relatively complete, and thus lengthy, summary of the comments at the meeting. The beginning of the summary sets out as conclusions some issues on which there appeared to be widespread agreement, but those conclusions are not intended to serve as a set of detailed recommendations. The meeting organizer is distributing his recommendations in a separate memorandum; recommendations from others who attended the meeting are welcome and will be distributed by the meeting organizer to the participants and to the Project committee.

NONE

1993-12-31

237

Human SCARB2-Mediated Entry and Endocytosis of EV71  

PubMed Central

Enterovirus (EV) 71 infection is known to cause hand-foot-and-mouth disease (HFMD) and in severe cases, induces neurological disorders culminating in fatality. An outbreak of EV71 in South East Asia in 1997 affected over 120,000 people and caused neurological disorders in a few individuals. The control of EV71 infection through public health interventions remains minimal and treatments are only symptomatic. Recently, human scavenger receptor class B, member 2 (SCARB2) has been reported to be a cellular receptor of EV71. We expressed human SCARB2 gene in NIH3T3 cells (3T3-SCARB2) to study the mechanisms of EV71 entry and infection. We demonstrated that human SCARB2 serves as a cellular receptor for EV71 entry. Disruption of expression of SCARB2 using siRNAs can interfere EV71 infection and subsequent inhibit the expression of viral capsid proteins in RD and 3T3-SCARB2 but not Vero cells. SiRNAs specific to clathrin or dynamin or chemical inhibitor of clathrin-mediated endocytosis were all capable of interfering with the entry of EV71 into 3T3-SCARB2 cells. On the other hand, caveolin specific siRNA or inhibitors of caveolae-mediated endocytosis had no effect, confirming that only clathrin-mediated pathway was involved in EV71 infection. Endocytosis of EV71 was also found to be pH-dependent requiring endosomal acidification and also required intact membrane cholesterol. In summary, the mechanism of EV71 entry through SCARB2 as the receptor for attachment, and its cellular entry is through a clathrin-mediated and pH-dependent endocytic pathway. This study on the receptor and endocytic mechanisms of EV71 infection is useful for the development of effective medications and prophylactic treatment against the enterovirus. PMID:22272359

Tsou, Yueh-Liang; Chitra, Ebenezer; Hsiao, Kuang-Nan; Shao, Hsiao-Yun; Liu, Chia-Chyi; Sia, Charles; Chong, Pele; Chow, Yen-Hung

2012-01-01

238

J Neuropathol Exp Neurol . Author manuscript Differential distribution of erbB receptors in human glioblastoma  

E-print Network

in human glioblastoma multiforme: expression of erbB3 in CD133-positive putative cancer stem cells V Glioblastomas are the most common CNS tumors in adults, and they remain resistant to current treatments. ErbB1 is a promising target for molecular therapy. However, data on erbB signaling in glioblastomas are still sparse

Paris-Sud XI, Université de

239

[Studies on the antitumor activity of human recombinant interferon alpha-2b in vitro].  

PubMed

Correlation between antiproliferative and binding activities of interferon (IFN) alpha-2b to various human cell lines was examined using human recombinant IFN alpha-2b. Burkitt's lymphoma Daudi cells and human renal cell carcinoma OS-RC-2 cells were sensitive to IFN alpha-2b, whereas two EB-virus-transformed B cell lines, FS and L-KT3, and human A375 melanoma cells showed low or no sensitivity. 125I-IFN alpha-2b binding assay revealed that the difference in IFN alpha-2b sensitivity was related to the number and the affinity of IFN alpha-2b receptors per cell. Experiments were then performed to investigate the influence of recombinant IFN alpha-2b on the cytostatic activity of monocytes against A375 cells in vitro. IFN alpha-2b enhanced the cytostatic activity of monocytes against A375 cells which showed low sensitivity to the direct growth inhibitory effect of IFN alpha-2b. Depletion of NK cells from the monocyte preparations by anti-Leu-11b monoclonal antibody and complement did not affect the monocyte activation by IFN alpha-2b, indicating that NK cells were not involved in this system. PMID:2946265

Haraguchi, S; Koide, Y; Matsuo, T; Yoshida, T O

1986-10-01

240

Single-step immunoaffinity purification and characterization of dodecylmaltoside-solubilized human neutrophil flavocytochrome b  

Microsoft Academic Search

Flavocytochrome b (Cyt b) is a heterodimeric, integral membrane protein that serves as the central component of an electron transferase system employed by phagocytes for elimination of bacterial and fungal pathogens. This report describes a rapid and efficient single-step purification of Cyt b from human neutrophil plasma membranes by solubilization in the nonionic detergent dodecylmaltoside (DDM) and immunoaffinity chromatography. A

Ross M Taylor; James B Burritt; Thomas R Foubert; Meagan A Snodgrass; Kim C Stone; Danas Baniulis; Jeannie M Gripentrog; Connie Lord; Algirdas J Jesaitis

2003-01-01

241

Human vitamin B12 absorption measurement by accelerator mass spectrometry using specifically  

E-print Network

Human vitamin B12 absorption measurement by accelerator mass spectrometry using specifically to assimilate dietary vitamin B12. Assaying and understanding absorption and uptake of B12 is important because and showed plasma appearance and clearance curves consistent with the predicted behavior of the pure vitamin

California at Davis, University of

242

Invasin of Yersinia pseudotuberculosis activates human peripheral B cells.  

PubMed Central

The Yersinia pseudotuberculosis cell surface-located protein invasin was found to promote binding between the pathogen and resting peripheral B cells via beta 1 integrin receptors (CD29). B cells responded by expressing several activation markers and by growing, In contrast, T cells did not react, although these cells express CD29. An isogenic invA mutant failed to activate B cells. The mutation could be complemented by providing the invA+ gene in trans. Purified invasin alone did not activate B cells, although it was able to block the binding of bacteria to the cells. PMID:8641788

Lundgren, E; Carballeira, N; Vazquez, R; Dubinina, E; Branden, H; Persson, H; Wolf-Watz, H

1996-01-01

243

Eur J Immunol. Author manuscript Splenic marginal zone B cells in humans: where do they mutate their Ig  

E-print Network

Animals ; Antigens, CD27 ; immunology ; B-Lymphocyte Subsets ; cytology ; immunology ; Cell Lineage ; genetics ; Mutation ; Spleen ; cytology ; immunology Introduction: Memory B cell subsets in humans" " R. K

Boyer, Edmond

244

Human Apolipoprotein B RNA Editing Deaminase Gene (APOBEC1)  

Microsoft Academic Search

Genomic clones encoding the human APOBEC1 gene and its 5? flanking region have been isolated and characterized. The human gene contains five coding exons. The introns dividing these exons correspond exactly to those found in the mouse gene. The translation initiation site, ATG, is located in exon 2 at the same site as in the mouse. The 5? flanking sequence

Takahiro Fujino; Naveenan Navaratnam; James Scott

1998-01-01

245

Inhibition of Nuclear Factor-Kappa B Activation Decreases Survival of Mycobacterium tuberculosis in Human Macrophages  

PubMed Central

Nuclear factor-kappa B (NF?B) is a ubiquitous transcription factor that mediates pro-inflammatory responses required for host control of many microbial pathogens; on the other hand, NF?B has been implicated in the pathogenesis of other inflammatory and infectious diseases. Mice with genetic disruption of the p50 subunit of NF?B are more likely to succumb to Mycobacterium tuberculosis (MTB). However, the role of NF?B in host defense in humans is not fully understood. We sought to examine the role of NF?B activation in the immune response of human macrophages to MTB. Targeted pharmacologic inhibition of NF?B activation using BAY 11-7082 (BAY, an inhibitor of I?B? kinase) or an adenovirus construct with a dominant-negative I?B? significantly decreased the number of viable intracellular mycobacteria recovered from THP-1 macrophages four and eight days after infection. The results with BAY were confirmed in primary human monocyte-derived macrophages and alveolar macrophages. NF?B inhibition was associated with increased macrophage apoptosis and autophagy, which are well-established killing mechanisms of intracellular MTB. Inhibition of the executioner protease caspase-3 or of the autophagic pathway significantly abrogated the effects of BAY. We conclude that NF?B inhibition decreases viability of intracellular MTB in human macrophages via induction of apoptosis and autophagy. PMID:23634218

Chmura, Kathryn; Ovrutsky, Alida R.; Su, Wen-Lin; Griffin, Laura; Pyeon, Dohun; McGibney, Mischa T.; Strand, Matthew J.; Numata, Mari; Murakami, Seiji; Gaido, Loretta; Honda, Jennifer R.; Kinney, William H.; Oberley-Deegan, Rebecca E.; Voelker, Dennis R.; Ordway, Diane J.; Chan, Edward D.

2013-01-01

246

Cucurbitacin B inhibits human breast cancer cell proliferation through disruption of microtubule polymerization and nucleophosmin/B23 translocation  

PubMed Central

Background Cucurbitacin B, an oxygenated tetracyclic triterpenoid compound extracted from the Thai medicinal plant Trichosanthes cucumerina L., has been reported to have several biological activities including anti-inflammatory, antimicrobial and anticancer. Cucurbitacin B is great of interest because of its biological activity. This agent inhibits growth of various types of human cancer cells lines. Methods In this study, we explored the novel molecular response of cucurbitacin B in human breast cancer cells, MCF-7 and MDA-MB-231. The growth inhibitory effect of cucurbitacin B on breast cancer cells was assessed by MTT assay. The effects of cucurbitacin B on microtubules morphological structure and tubulin polymerization were analyzed using immunofluorescence technique and tubulin polymerization assay kit, respectively. Proteomic analysis was used to identify the target-specific proteins that involved in cucurbitacin B treatment. Some of the differentially expressed genes and protein products were validated by real-time RT-PCR and western blot analysis. Cell cycle distributions and apoptosis were investigated using flow cytometry. Results Cucurbitacin B exhibited strong antiproliferative effects against breast cancer cells in a dose-dependent manner. We show that cucurbitacin B prominently alters the cytoskeletal network of breast cancer cells, inducing rapid morphologic changes and improper polymerization of the microtubule network. Moreover, the results of 2D-PAGE, real-time RT-PCR, and western blot analysis revealed that the expression of nucleophosmin/B23 and c-Myc decreased markedly after cucurbitacin B treatment. Immunofluorescence microscopy showed that cucurbitacin B induced translocation of nucleophosmin/B23 from the nucleolus to nucleoplasm. Treatment with cucurbitacin B resulted in cell cycle arrest at G2/M phase and the enhancement of apoptosis. Conclusions Our findings suggest that cucurbitacin B may inhibit the proliferation of human breast cancer cells through disruption of the microtubule network and down-regulation of c-Myc and nucleophosmin/B23 as well as the perturbation in nucleophosmin/B23 trafficking from the nucleolus to nucleoplasm, resulting in G2/M arrest. PMID:23062075

2012-01-01

247

The signs B and B-bent in Israeli sign language according to the theory of Phonology as Human Behavior.  

PubMed

The purpose of the present research is to examine which of the two factors: (1) the iconic-semiotic factor; or (2) the human-phonetic factor is more relevant in explaining the appearance and distribution of the hand shape B-bent in Israeli Sign Language (ISL). The B-bent shape has been the subject of much attention in sign language research revolving around the question of its status as a phoneme. The arguments supporting the phonemic status of the B-bent hand shape have been primarily based on the semiotic opposition between the hand shape B and the hand shape B-bent. It has been claimed that in Italian Sign Language the hand shape B is perceptually distinct from the hand shape B-bent, i.e. in opposition to the general, neutral, unmarked meaning of the hand shape B, the iconic hand shape B-bent has a more narrow, specific and marked meaning: DELIMIT. The B-bent hand shape appears in spatial-temporal signs such as "a little before, ahead, postpone or behind". In these signs the iconic structure of the hand shape B-bent is utilized to mark borders in space and time. The arguments opposing the perceptual/phonemic distinction between these hand shapes is based on the human-phonetic factor, i.e. the need to reduce the effort on the part of the wrist joints in specific phonetic environments. We performed a quantitative and qualitative content analysis of the distribution of the basic units of 560 lexical signs taken from a stratified random sample from the ISL dictionary. The results were analyzed in the framework of the sign-oriented linguistic theory of the Columbia School including the theory of Phonology as Human Behavior. Our data revealed that the B-bent hand shape--as all the "building blocks" of the ISL--is a morpho-phonemic unit. We found that there is not only a phonemic distinction between hand shape B and hand shape B-bent in ISL (based on minimal pairs), but there is also a perceptual distinction between them. The qualitative analysis shows that the distribution of the B and the B-bent shapes in the lexicon of ISL is not random but rather motivated by a specific iconic meaning attached to each one of them. The results of our research lead to the conclusion that iconicity is the dominant explanatory factor in the Phonology of ISL. PMID:18415739

Fuks, Orit; Tobin, Yishai

2008-01-01

248

Draft Genome Sequences of Two Pairs of Human Intestinal Bifidobacterium longum subsp. longum Strains, 44B and 1-6B and 35B and 2-2B, Consecutively Isolated from Two Children after a 5-Year Time Period  

PubMed Central

We report the genome sequences of four isolates of a human gut symbiont, Bifidobacterium longum. Strains 44B and 35B were isolated from two 1-year-old infants, while 1-6B and 2-2B were isolated from the same children 5years later. The sequences permit investigations of factors enabling long-term colonization of bifidobacteria. PMID:23682142

Shkoporov, A. N.; Efimov, B. A.; Khokhlova, E. V.; Chaplin, A. V.; Kafarskaya, L. I.; Durkin, A. S.; McCorrison, J.; Torralba, M.; Gillis, M.; Sutton, G.; Weibel, D. B.; Nelson, K. E.

2013-01-01

249

Effects of Clostridium difficile toxin B on human monocytes and macrophages: possible relationship with cytoskeletal rearrangement.  

PubMed Central

Toxin B from Clostridium difficile is cytopathic in vitro for various types of cells, including polymorphonuclear cells, lymphocytes, and monocytes. Since intestine lamina propria is rich in macrophages, we studied the effect of toxin B on human monocytes and on human macrophages generated in vitro by long-term culture of purified circulating blood monocytes. Upon addition of toxin B, human monocytes exhibited few modifications whereas macrophages adopted a stellate morphology, with rounding up of the perikaryon. Toxin B made microfilaments of actin disappear and induced an important reorganization of vimentin and a redistribution of tubulin. Membrane area increased by approximately 16%. Toxin B did not affect the viability of human mononuclear phagocytes and did not exert any significant lytic effect. It profoundly altered the phagocytic function of macrophages. When activated by gamma interferon in the presence of toxin B, monocytes were more cytotoxic for U-937 target cells than control monocytes activated in absence of toxin. Finally, the combined treatment of monocytes with gamma interferon and toxin B increased significantly the secretion of tumor necrosis factor alpha, whereas toxin B alone was unable to induce tumor necrosis factor production. These results suggest that morphological and functional alterations induced in human mononuclear phagocytes by toxin B from C. difficile are due to the disorganization of the cytoskeleton and the resulting impairment of the membrane traffic equilibrium. Images PMID:8432590

Siffert, J C; Baldacini, O; Kuhry, J G; Wachsmann, D; Benabdelmoumene, S; Faradji, A; Monteil, H; Poindron, P

1993-01-01

250

Chromosomal localization of the gene for human B-cell antigen CD40  

SciTech Connect

CD40 is a surface glycoprotein expressed on all human B lymphocytes and plays an important role in B-cell development, growth, and differentiation. Anti-CD40 monoclonal antibodies cause isotype switching in B cells treated with IL-4. CD40 is a member of a family of proteins that include low-affinity nerve growth factor receptor, TNF receptor, and the antigen Fas. The ligand for CD40 had been recently identified and had been assigned to the X chromosome. Using a panel of human-rodent somatic cell hybrids, the authors now show that CD40 maps to human chromosome 20.

Ramesh, N.; Geha, R. (Harvard Medical School, Boston, MA (United States)); Ramesh, V.; Gusella, J.F. (Massachusetts General Hospital, Boston (United States))

1993-05-01

251

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.

2010-01-01

252

Enterotoxin A and B production in strains of Staphylococcus aureus isolated from human beings and foods.  

PubMed Central

The production of enterotoxin A and B by strains of Staphylococcus aureus isolated from nasal swabs of healthy carriers, from lesions of hospital patients and from foods unconnected with outbreaks of food-poisoning was investigated. Sixty-six strains of S. aureus were obtained from human beings, two produced enterotoxin A, 45 produced enterotoxin B, seven produced enterotoxins A + B. Thirty-six strains were isolated from 111 samples of food, one produced enterotoxin A, 16 produced enterotoxin B. The relative incidence of A, B and A + B enterotoxigenicity was assessed. PMID:7057022

Reali, D.

1982-01-01

253

Quantitative Genomic and Antigenomic Enterovirus RNA Detection in Explanted Heart Tissue Samples from Patients with End-Stage Idiopathic Dilated Cardiomyopathy  

PubMed Central

Standardized one-step real-time RT-PCR assay detected enterovirus RNA in cardiac biopsy samples from 4 of 20 patients suffering from idiopathic dilated cardiomyopathy (IDCM). The median viral load was 287 copies per microgram of total extracted nucleic acids, with positive- to negative-strand RNA ratios ranging from 2 to 20. These results demonstrate enterovirus persistence in the heart of IDCM patients, characterized by low viral loads and low positive- to negative-RNA ratios. PMID:22837323

Renois, Fanny; Talmud, Deborah; Nguyen, Yohan; Lesaffre, Francois; Boulagnon, Camille; Bruneval, Patrick; Fornes, Paul; Andreoletti, Laurent

2012-01-01

254

Detection of antigens specific for B-lymphoid cultured cell lines with human alloantisera  

PubMed Central

Human sera were tested for cytotoxicity to pairs of long-term tissue- cultured cell lines. Each pair had been derived from the same individual and one of the pairs possessed the characteristics of either "T" or "B" cells. The alloantisera used were HL-A-typing reagents or sera obtained from Amish multiparas. Selected cytotoxicity was found against the B-cell lines by direct testing. Cytotoxicity was abolished by absorption with B-cell line but not by absorption with the T-cell lines. The results suggest that a group of allotypic antigens may be expressed exculsively on human B cells. PMID:1080182

1975-01-01

255

Flotillin depletion affects ErbB protein levels in different human breast cancer cells.  

PubMed

The ErbB3 receptor is an important regulator of cell growth and carcinogenesis. Among breast cancer patients, up to 50-70% have ErbB3 overexpression and 20-30% show overexpressed or amplified ErbB2. ErbB3 has also been implicated in the development of resistance to several drugs used against cancers driven by ErbB1 or ErbB2. One of the main challenges in ErbB-targeting therapy is to inactivate signaling mediated by ErbB2-ErbB3 oncogenic receptor complexes. We analyzed the regulatory role of flotillins on ErbB3 levels and ErbB2-ErbB3 complexes in SKBR3, MCF7 and MDA-MB-134-VI human breast cancer cells. Recently, we described a mechanism for interfering with ErbB2 signaling in breast cancer and demonstrated a molecular complex of flotillin scaffolding proteins with ErbB2 and Hsp90. In the present study, flotillins were found to be in a molecular complex with ErbB3, even in cells without the presence of ErbB2 or other ErbB receptors. Depletion of either flotillin-1 or flotillin-2 resulted in downregulation of ErbB3 and a selective reduction of ErbB2-ErbB3 receptor complexes. Moreover, flotillin-2 depletion resulted in reduced activation of Akt and MAPK signaling cascades, and as a functional consequence of flotillin depletion, breast cancer cells showed an impaired cell migration. Altogether, we provide data demonstrating a novel and functional role of flotillins in the regulation of ErbB protein levels and stabilization of ErbB2-ErbB3 receptor complexes. Thus, flotillins are crucial regulators for oncogenic ErbB function and potential targets for cancer treatment. PMID:24747692

Asp, Nagham; Pust, Sascha; Sandvig, Kirsten

2014-09-01

256

Characterization of human b,b-carotene-15,150 -monooxygenase  

E-print Network

, leads to blindness in hundreds of thousands of children annually, as well as great in- creases in childhood morbidity [7]. The amount of vitamin A obtainable from dietary b,b-carotene depends mainly on two

Palczewski, Krzysztof

257

Toward discovery science of human brain function Bharat B. Biswala  

E-print Network

Hampsoni , Matthew J. Hoptmanj , James S. Hydek , Vesa J. Kiviniemil , Rolf Kötterm , Shi-Jiang Lin , Ching , James J. Pekarx , Scott J. Peltiery , Steven E. Petersenz , Valentin Riedlaa , Serge A. R. B. Romboutsbb

Miall, Chris

258

A feeder-free differentiation system identifies autonomously proliferating B cell precursors in human bone marrow.  

PubMed

The peripheral B cell compartment is maintained by homeostatic proliferation and through replenishment by bone marrow precursors. Because hematopoietic stem cells cycle at a slow rate, replenishment must involve replication of precursor B cells. To study proliferation of early human B cell progenitors, we established a feeder cell-free in vitro system allowing the development of B cells from CD34(+) hematopoietic stem cells up to the stage of immature IgM(+) B cells. We found that pro-B and pre-B cells generated in vitro can proliferate autonomously and persist up to 7 wk in culture in the absence of signals induced by exogenously added cytokines. Nevertheless, addition of IL-7 enhanced pre-B cell expansion and inhibited maturation into IgM(+) B cells. The B cell precursor subsets replicating in vitro were highly similar to the bone marrow B cell precursors cycling in vivo. The autonomous proliferation of B cell precursor subsets in vitro and their long-term persistence implies that proliferation during pro-B and pre-B cell stages plays an important role in the homeostasis of the peripheral B cell compartment. Our in vitro culture can be used to study defects in B cell development or in reconstitution of the B cell pool after depletion and chemotherapy. PMID:24379121

Kraus, Helene; Kaiser, Sandra; Aumann, Konrad; Bnelt, Peter; Salzer, Ulrich; Vestweber, Dietmar; Erlacher, Miriam; Kunze, Mirjam; Burger, Meike; Pieper, Kathrin; Sic, Heiko; Rolink, Antonius; Eibel, Hermann; Rizzi, Marta

2014-02-01

259

The use of MR B+1 imaging for validation of FDTD electromagnetic simulations of human anatomies  

NASA Astrophysics Data System (ADS)

In this study, MR B+1 imaging is employed to experimentally verify the validity of FDTD simulations of electromagnetic field patterns in human anatomies. Measurements and FDTD simulations of the B+1 field induced by a 3 T MR body coil in a human corpse were performed. It was found that MR B+1 imaging is a sensitive method to measure the radiofrequency (RF) magnetic field inside a human anatomy with a precision of approximately 3.5%. A good correlation was found between the B+1 measurements and FDTD simulations. The measured B+1 pattern for a human pelvis consisted of a global, diagonal modulation pattern plus local B+1 heterogeneties. It is believed that these local B+1 field variations are the result of peaks in the induced electric currents, which could not be resolved by the FDTD simulations on a 5 mm3 simulation grid. The findings from this study demonstrate that B+1 imaging is a valuable experimental technique to gain more knowledge about the dielectric interaction of RF fields with the human anatomy.

Van den Berg, Cornelis A. T.; Bartels, Lambertus W.; van den Bergen, Bob; Kroeze, Hugo; de Leeuw, Astrid A. C.; Van de Kamer, Jeroen B.; Lagendijk, Jan J. W.

2006-10-01

260

B7-H5 costimulates human T cells via CD28H  

E-print Network

The B7/CD28 family has profound modulatory effects in immune responses and constitutes an important target for the development of novel therapeutic drugs against human diseases. Here we describe a new CD28 homologue (CD28H) ...

Zhu, Yuwen

261

Human Resource Services -Payroll 1265 Military Trail, Room B526C/D  

E-print Network

(Dec 2012) Human Resource Services - Payroll 1265 Military Trail, Room B526C/D Toronto, Ontario, M1 # Of Months Total for this contract Hours Worked x = Salary $ x = $ TOTAL SALARY (EXCLUDING Vacation Pay

Kronzucker, Herbert J.

262

Human Resource Services -Payroll 1265 Military Trail, Room B526C/D  

E-print Network

(Dec 2011) Human Resource Services - Payroll 1265 Military Trail, Room B526C/D Toronto, Ontario, M1. George Campus: Tenure Salary Stipend Monthly Salary/Stipend: Total Salary/Stipend: Percent of time (for

Kronzucker, Herbert J.

263

Human Resource Services -Payroll 1265 Military Trail, Room B526C/D  

E-print Network

(Dec 2013) Human Resource Services - Payroll 1265 Military Trail, Room B526C/D Toronto, Ontario, M1. George Campus: Tenure Salary Stipend Monthly Salary/Stipend: Total Salary/Stipend: Percent of time (for

Boonstra, Rudy

264

Human Resource Services -Payroll 1265 Military Trail, Room B526C/D  

E-print Network

(Dec 2013) Human Resource Services - Payroll 1265 Military Trail, Room B526C/D Toronto, Ontario, M1 # Of Months Total for this contract Hours Worked x = Salary $ x = $ TOTAL SALARY (EXCLUDING Vacation Pay

Boonstra, Rudy

265

Solving Hex: Beyond Humans Broderick Arneson, Ryan B. Hayward, Philip Henderson  

E-print Network

Solving Hex: Beyond Humans Broderick Arneson, Ryan B. Hayward, Philip Henderson Dept. of Computing, CG2010 (Kanazawa), H.J. van den Herik and H. Iida and A. Plaat (eds.), Springer LNCS 6515, 2011, pp 1

Hayward, Ryan B.

266

Omalizumab may decrease IgE synthesis by targeting membrane IgE+ human B cells  

E-print Network

Omalizumab, is a humanized anti-IgE monoclonal antibody used to treat allergic asthma. Decreased serum IgE levels, lower eosinophil and B cell counts have been noted as a result of treatment. In vitro studies and animal ...

Chan, Marcia A.; Gigliotti, Nicole M.; Dotson, Abby Louise; Rosenwasser, Lanny J.

2013-09-02

267

Identification of LMX1B as a novel oncogene in human ovarian cancer.  

PubMed

Ovarian cancers are thought to result from the accumulation of multiple genetic aberrations that transform ovarian and/or fallopian tube surface epithelial cells, allowing for their abnormal growth, proliferation and metastasis. In the report presented here, we carried out genome-wide copy-number analysis using comparative genomic hybridization on a panel of mouse ovarian cancer (OVCA) cell lines previously established in our laboratory. We identified a recurrent focal amplification on mouse chromosomal region 2qB, which contains the LIM-homeodomain-containing transcription factor 1B (Lmx1b) gene. LMX1B is not expressed in normal human ovary, but is expressed in many human OVCA cell lines and primary tumors. High expression of LMX1B correlates with poor outcome. To clarify the role of LMX1B in ovarian carcinogenesis, we transduced LMX1B into a panel of mouse and human OVCA cell lines and demonstrated that LMX1B strongly promotes migration of cancer cells in culture and promotes xenograft growth in nude mice. Conversely, knockdown of LMX1B in a human cell line with endogenous high expression of LMX1B inhibits cell migration in vitro and tumor growth in vivo. Microarray analysis of cells overexpressing LMX1B identified the nuclear factor (NF)-?B pathway as a potential mediator of tumor progression and subsequent treatment of NF?B inhibitor decreased the migratory capacity of these cells. Thus, our data demonstrate that LMX1B is a novel oncogene in OVCA pathogenesis. PMID:24056967

He, L; Guo, L; Vathipadiekal, V; Sergent, P A; Growdon, W B; Engler, D A; Rueda, B R; Birrer, M J; Orsulic, S; Mohapatra, G

2014-08-14

268

Anti-Enterovirus 71 Effects of Chrysin and Its Phosphate Ester  

PubMed Central

Enterovirus 71 (EV71) can cause severe disease and even lead to death in children, and an effective antiviral drug is currently unavailable. The anti-EV71 effect of chrysin (5,7-dihydroxyflavone), a natural flavonoid commonly found in many plants, was tested in this report. By using the predicting program Autodock 4.0 and an in vitro protease inhibition assay, we found that chrysin could suppress viral 3Cpro activity. Replication of viral RNA and production of viral capsid protein and the infectious virion were strongly inhibited by chrysin, without noticeable cytotoxicity. Cytopathic effects on cells were also prevented. Diisopropyl chrysin-7-yl phosphate (CPI), the phosphate ester for chrysin, was generated through a simplified Atheron-Todd reaction to achieve stronger anti-viral activity. CPI was also able to bind with and inhibit viral 3Cpro activity in vitro. As expected, CPI demonstrated more potent antiviral activity against EV71. PMID:24598537

Du, Jiang; Cui, Sheng; Yang, Fan; Jin, Qi

2014-01-01

269

The interleukin-12 and interleukin-12 receptor system in normal and transformed human B-lymphocytes  

Microsoft Academic Search

Background and Objectives. Interleukin-12 (IL-12) is a heterodimeric cytokine that induces interferon- ? (IFN-?) production by natural killer and T-lympho- cytes. IL-12 also activates human B-cells through the IL-12 receptor (IL-12R) complex. Here we review the expression and function of IL-12 and IL-12R in human B-cells and in their malignant counterparts. Evidence and Information Sources. The information provided derives from

IRMA AIROLDI; ROBERTA GUGLIELMINO; GIUSEPPE CARRA; ANNA CORCIONE; FRANCA GEROSA; GIUSEPPE TABORELLI; GIORGIO TRINCHIERI; VITO PISTOIA

270

In Vivo Time-Related Evaluation of a Therapeutic Neutralization Monoclonal Antibody against Lethal Enterovirus 71 Infection in a Mouse Model  

PubMed Central

Enterovirus 71 (EV71) is a neurotropic virus capable of inducing severe neurological symptoms and death. No direct targeting antivirals are useful in the treatment of severe EV71 infection. Because of low toxicity and good specificity, monoclonal antibodies (MAb) are a potential candidate for the treatment of viral infections. Therefore, we developed an EV71-specific conformational MAb with high in vitro cross-neutralization activity to heterologous EV71 subgenotypes. The in vivo treatment experiment at different days post-infection indicated that a single treatment of MAb CT11F9 within day 3 post-infection fully protected mice from morbidity and mortality (0% PBS vs. 100% at 10 g/g per body weight ***P<0.0001). Immunohistochemical and histological analysis confirmed that CT11F9 significantly prohibited EV71 VP1 expression in various tissues and prevented EV71-induced myonecrosis. Moreover, thrice-treatment at day 4, 5, 6 post-infection was associated with an increased survival rate (18.2% single vs. 50% thrice at 20 g/g per body weight), and the mice recovered from limb paralysis. Competitive ELISA also confirmed that CT11F9-recognized epitopes were immunodominant in humans. In conclusion, MAb CT11F9 is an ideal candidate to be humanized and used in severe EV71 infection. PMID:25279734

Li, Zhiqun; Xu, Longfa; He, Delei; Yang, Lisheng; Liu, Che; Chen, Yixin; Shih, James Wai Kuo; Zhang, Jun; Zhao, Qinjian; Cheng, Tong; Xia, Ningshao

2014-01-01

271

Blood-brain transfer of Pittsburgh compound B in humans  

PubMed Central

In the labeled form, the Pittsburgh compound B (2-(4?-{N-methyl-[11C]}methyl-aminophenyl)-6-hydroxy-benzothiazole, [11C]PiB), is used as a biomarker for positron emission tomography (PET) of brain ?-amyloid deposition in Alzheimer's disease (AD). The permeability of [11C]PiB in the blood-brain barrier is held to be high but the permeability-surface area product and extraction fractions in patients or healthy volunteers are not known. We used PET to determine the clearance associated with the unidrectional blood-brain transfer of [11C]PiB and the corresponding cerebral blood flow rates in frontal lobe, whole cerebral cortex, and cerebellum of patients with Alzheimer's disease and healthy volunteers. Regional cerebral blood flow rates differed significantly between the two groups. Thus, regional and whole-brain permeability-surface area products were identical, in agreement with the observation that numerically, but insignificantly, unidirectional blood-brain clearances are lower and extraction fractions higher in the patients. The evidence of unchanged permeability-surface area products in the patients implies that blood flow changes can be deduced from the unidirectional blood-brain clearances of [11C]PiB in the patients. PMID:24223554

Gjedde, Albert; Aanerud, Joel; Braendgaard, Hans; Rodell, Anders B.

2013-01-01

272

BDNF and its TrkB receptor in human fracture healing.  

PubMed

Fracture healing is a physiological process of repair which proceeds in stages, each characterized by a different predominant tissue in the fracture gap. Matrix reorganization is regulated by cytokines and growth factors. Neurotrophins and their receptors might be of importance to osteoblasts and endothelial cells during fracture healing. The aim of this study was to examine the presence of brain-derived neurotrophic factor (BDNF) and its tropomyosin-related kinase B receptor (TrkB) during human fracture healing. BDNF and TrkB were investigated in samples from human fracture gaps and cultured cells using RT-PCR, Western blot, and immunohistochemistry. Endothelial cells and osteoblastic cell lines demonstrated a cytoplasmic staining pattern of BDNF and TrkB in vitro. At the mRNA level, BDNF and TrkB were expressed in the initial and osteoid formation phase of human fracture healing. In the granulation tissue of fracture gap, both proteins--BDNF and TrkB--are concentrated in endothelial and osteoblastic cells at the margins of woven bone suggesting their involvement in the formation of new vessels. There was no evidence of BDNF or TrkB during fracture healing in chondrocytes of human enchondral tissue. Furthermore, BDNF is absent in mature bone. Taken together, BDNF and TrkB are involved in vessel formation and osteogenic processes during human fracture healing. The detection of BDNF and its TrkB receptor during various stages of the bone formation process in human fracture gap tissue were shown for the first time. The current study reveals that both proteins are up-regulated in human osteoblasts and endothelial cells in fracture healing. PMID:24984919

Kilian, Olaf; Hartmann, Sonja; Dongowski, Nicole; Karnati, Srikanth; Baumgart-Vogt, Eveline; Hrtel, Frauke V; Noll, Thomas; Schnettler, Reinhard; Lips, Katrin Susanne

2014-09-01

273

An AICDA-independent mechanism of secondary VH gene rearrangement in preimmune human B cells Running title: Secondary rearrangement in human B cells  

PubMed Central

VH replacement is a form of IgH chain receptor editing that is believed to be mediated by recombinase cleavage at cryptic recombination signals (cRSS) embedded in IGHV genes. Whereas there are several reports of IGHV replacement in primary and transformed human B cells and murine models, it remains unclear whether IGHV replacement contributes to the normal human B cell repertoire. We identified VH ? VHDJH compound rearrangements from fetal liver, fetal bone marrow and naive peripheral blood, all of which involved invading and recipient IGHV4 genes that contain a cryptic heptamer, 13 base pair (bp) spacer and nonamer in the 5' portion of framework region (FR) 3. Surprisingly, all pseudohybrid joins lacked molecular processing associated with typical VHDJH recombination or nonhomologous end joining. Although inefficient compared to a canonical RSS, the IGHV4 cRSS was a significantly better substrate for in vitro RAG-mediated cleavage than the IGHV3 cRSS. It has been suggested that activation induced cytidine deamination (AICDA) may contribute to VH replacement. However, we found similar secondary rearrangements utilizing IGHV4 genes in AICDA-deficient human B cells. The data suggest that IGHV4 replacement in preimmune human B cells is mediated by an AICDA-independent mechanism resulting from inefficient but selective RAG activity. PMID:19017972

Longo, Nancy S.; Grundy, Gabrielle J.; Lee, Jisoo; Gellert, Martin; Lipsky, Peter E.

2008-01-01

274

Harnessing Gene Conversion in Chicken B Cells to Create a Human Antibody Sequence Repertoire  

PubMed Central

Transgenic chickens expressing human sequence antibodies would be a powerful tool to access human targets and epitopes that have been intractable in mammalian hosts because of tolerance to conserved proteins. To foster the development of the chicken platform, it is beneficial to validate transgene constructs using a rapid, cell culture-based method prior to generating fully transgenic birds. We describe a method for the expression of human immunoglobulin variable regions in the chicken DT40 B cell line and the further diversification of these genes by gene conversion. Chicken VL and VH loci were knocked out in DT40 cells and replaced with human VK and VH genes. To achieve gene conversion of human genes in chicken B cells, synthetic human pseudogene arrays were inserted upstream of the functional human VK and VH regions. Proper expression of chimeric IgM comprised of human variable regions and chicken constant regions is shown. Most importantly, sequencing of DT40 genetic variants confirmed that the human pseudogene arrays contributed to the generation of diversity through gene conversion at both the Igl and Igh loci. These data show that engineered pseudogene arrays produce a diverse pool of human antibody sequences in chicken B cells, and suggest that these constructs will express a functional repertoire of chimeric antibodies in transgenic chickens. PMID:24278246

Schusser, Benjamin; Yi, Henry; Collarini, Ellen J.; Izquierdo, Shelley Mettler; Harriman, William D.; Etches, Robert J.; Leighton, Philip A.

2013-01-01

275

Development and Evaluation of EPA Method 1615 for Detection of Enterovirus and Norovirus in Water  

PubMed Central

The U.S. EPA developed a sample concentration and preparation assay in conjunction with the total culturable virus assay for concentrating and measuring culturable viruses in source and drinking waters as part of the Information Collection Rule (ICR) promulgated in 1996. In an effort to improve upon this method, the U.S. EPA recently developed Method 1615: Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Method 1615 uses a culturable virus assay with reduced equipment and labor costs compared to the costs associated with the ICR virus method and introduces a new molecular assay for the detection of enteroviruses and noroviruses by reverse transcription-quantitative PCR. In this study, we describe the optimization of several new components of the molecular assay and examine virus recovery from ground, reagent-grade, and surface water samples seeded with poliovirus type 3 and murine norovirus. For the culturable virus and molecular assays, mean poliovirus recovery using the complete method was 58% and 20% in groundwater samples, 122% and 39% using low-titer spikes in reagent-grade water, 42% and 48% using high-titer spikes in reagent-grade water, and 11% and 10% in surface water with high turbidity, respectively. Murine norovirus recovery by the molecular assay was 30% in groundwater samples, less than 8% in both low- and high-titer spikes in reagent-grade water, and 6% in surface water with high turbidity. This study demonstrates the effectiveness of Method 1615 for use with groundwater samples and highlights the need for further research into its effectiveness with surface water. PMID:23087037

Brinkman, Nichole E.; Griffin, Shannon M.; McMinn, Brian R.; Rhodes, Eric R.; Varughese, Eunice A.; Grimm, Ann C.; Parshionikar, Sandhya U.; Wymer, Larry; Fout, G. Shay

2013-01-01

276

Human Resource Development and Manpower Training. Paper Presentations: Session B.  

ERIC Educational Resources Information Center

This document contains 18 papers from the human resource development and manpower training section of an international conference on vocational education and training (VET) for lifelong learning in the information era. The following papers are included: "Use of Social and Economic Modeling to Plan Vocational Education and Training" (David L.

2000

277

Voltage-Gated Na+ channel ?1B: a secreted cell adhesion molecule involved in human epilepsy  

PubMed Central

Scn1b null mice have a severe neurological and cardiac phenotype. Human mutations in SCN1B result in epilepsy and cardiac arrhythmia. SCN1B is expressed as two developmentally regulated splice variants, ?1 and ?1B, that are each expressed in brain and heart in rodents and humans. Here we studied the structure and function of ?1B and investigated a novel human SCN1B epilepsy-related mutation (p.G257R) unique to ?1B. We show that wild-type ?1B is not a transmembrane protein, but a soluble protein expressed predominantly during embryonic development that promotes neurite outgrowth. Association of ?1B with voltage-gated Na+ channels (VGSCs) Nav1.1 or Nav1.3 is not detectable by immunoprecipitation and ?1B does not affect Nav1.3 cell surface expression as measured by [3H]-saxitoxin binding. However, ?1B co-expression results in subtle alteration of Nav1.3 currents in transfected cells, suggesting that ?1B may modulate Na+ current in brain. Similar to the previously characterized p.R125C mutation, p.G257R results in intracellular retention of ?1B, generating a functional null allele. In contrast, two other SCN1B mutations associated with epilepsy, p.C121W and p.R85H, are expressed at the cell surface. We propose that ?1B p.G257R may contribute to epilepsy through a mechanism that includes intracellular retention resulting in aberrant neuronal pathfinding. PMID:21994374

Patino, G.A.; Brackenbury, W.J.; Bao, Y.; Lopez-Santiago, L. F.; O'Malley, H.A.; Chen, C.; Calhoun, J.D.; Lafreniere, R.G.; Cossette, P.; Rouleau, G.A.; Isom, L.L.

2011-01-01

278

METABOLISM AND CYTOTOXICITY OF AFLATOXIN B 1 IN CYTOCHROME P-450EXPRESSING HUMAN LUNG CELLS  

Microsoft Academic Search

The mycotoxin aflatoxin B 1 (AFB 1 ) is a hepatocarcinogen in many animal models and probably a human carcinogen. Besides being a dietary carcinogen, AFB 1 has been detected in dusts generated in the processing and transportation of AFB 1 -contaminated products. Inhalation of grain dusts contaminated with AFB 1 may be a risk factor in human lung cancer.

Terry R. Van Vleet; Patrick J. Klein; Roger A. Coulombe Jr

2002-01-01

279

Gender Classification of Human Faces Arnulf B. A. Graf and Felix A. Wichmann  

E-print Network

Gender Classification of Human Faces Arnulf B. A. Graf and Felix A. Wichmann Max Planck Institute- chine (SVM) classification for a behaviorally important task in humans: gender classification, PCA, LLE, gender classification, SVM 1 Introduction Gender classification is arguably one of the more

280

The NADPH Metabolic Network Regulates Human aB-crystallin Cardiomyopathy and Reductive Stress in  

E-print Network

The NADPH Metabolic Network Regulates Human aB- crystallin Cardiomyopathy and Reductive Stress for a number of inherited human disorders, including cardiomyopathy, skeletal muscle myopathy, and cataracts. In a mouse model, cardiomyopathy caused by the dominant CryABR120G missense mutation was suppressed

281

UNIT 1B.3Phenotypic Analysis of Human Embryonic Stem Cells  

E-print Network

UNIT 1B.3Phenotypic Analysis of Human Embryonic Stem Cells Mark Ungrin,1 Michael O'Connor,2 Laboratory, BC Cancer Agency, Vancouver, British Columbia 3 StemCell Technologies, Inc, Vancouver, British Columbia ABSTRACT Human embryonic stem cells (hESCs) are an important tool for the study of develop- mental

Zandstra, Peter W.

282

Extending animal models of fear conditioning to humans M.R. Delgado a,*, A. Olsson b  

E-print Network

the extinction phase. This manuscript reviews how animal models of fear are translated to human behavior, and howExtending animal models of fear conditioning to humans M.R. Delgado a,*, A. Olsson b , E.A. Phelps, a simple paradigm that has been extensively investigated in animals, helping outline a brain circuitry

Phelps, Elizabeth

283

human-infective trypanosome T. b. rhodesi-ense has evolved a different way to escape  

E-print Network

human-infective trypanosome T. b. rhodesi- ense has evolved a different way to escape killing hypohaptoglobin- aemia) is endemic throughout sub-Saharan Africa. Instead, the parasite has evolved a pro- tein resistance to the TLFs present in most humans. SRA binds to APOL1 with high affinity and prevents either

McGehee, Michael

284

Dynamic Control of Human Eye on Head System Bijoy K. Ghosh1, Indika B. Wijayasinghe1  

E-print Network

Dynamic Control of Human Eye on Head System Bijoy K. Ghosh1, Indika B. Wijayasinghe1 1. Laboratory.ghosh, indika.wijayasinghe}@ttu.edu Abstract: In this paper we study the human eye movement, when the eye shifts is restrained to remain fixed, eye movements obey Listing's constraint, which states that the allowed

Ghosh, Bijoy K.

285

Comparative Gene Expression Profiling Identifies Common Molecular Signatures of NF-?B Activation in Canine and Human Diffuse Large B Cell Lymphoma (DLBCL)  

PubMed Central

We present the first comparison of global transcriptional changes in canine and human diffuse large B-cell lymphoma (DLBCL), with particular reference to the nuclear factor-kappa B (NF-?B) pathway. Microarray data generated from canine DLBCL and normal lymph nodes were used for differential expression, co-expression and pathway analyses, and compared with analysis of microarray data from human healthy and DLBCL lymph nodes. The comparisons at gene level were performed by mapping the probesets in canine microarrays to orthologous genes in humans and vice versa. A considerable number of differentially expressed genes between canine lymphoma and healthy lymph node samples were also found differentially expressed between human DLBCL and healthy lymph node samples. Principal component analysis using a literature-derived NF-?B target gene set mapped to orthologous canine array probesets and human array probesets clearly separated the healthy and cancer samples in both datasets. The analysis demonstrated that for both human and canine DLBCL there is activation of the NF-?B/p65 canonical pathway, indicating that canine lymphoma could be used as a model to study NF-?B-targeted therapeutics for human lymphoma. To validate this, tissue arrays were generated for canine and human NHL and immunohistochemistry was employed to assess NF-?B activation status. In addition, human and canine B-cell lymphoma lines were assessed for NF-?B activity and the effects of NF-?B inhibition. PMID:24023754

Mudaliar, Manikhandan A. V.; Haggart, Ross D.; Miele, Gino; Sellar, Grant; Tan, Karen A. L.; Goodlad, John R.; Milne, Elspeth; Vail, David M.; Kurzman, Ilene

2013-01-01

286

Hepatitis B Virus Infection in Cultured Human Lymphoblastoid Cells  

Microsoft Academic Search

Since it has been postulated that liver hepatocytes may become infected by hepatitis B virus (HBV) in vivo through direct contact with infected macrophages, the possibility that a circulating cell of hematopoietic origin might be susceptible to infection with HBV was investigated. Cells positive for HBV surface antigen were identified in aspirates of bone marrow cells from people infected with

Jean-Loup Romet-Lemonne; Mary Frances McLane; Emile Elfassi; William A. Haseltine; Jose Azocar; Myron Essex

1983-01-01

287

Functional characterization of novel mutations in the human cytochrome b gene  

Microsoft Academic Search

The great variability of the human mitochondrial DNA (mtDNA) sequence induces many difficulties in the search for its deleterious mutations. We illustrate these pitfalls by the analysis of the cytochrome b gene of 21 patients affected with a mitochondrial disease. Eighteen different sequence variations were found, five of which were new mutations. Extensive analysis of the cytochrome b gene of

Frdric Legros; Evi Chatzoglou; Paule Frachon; Hlne Ogier de Baulny; Pascal Lafort; Claude Jardel; Catherine Godinot; Anne Lombs

2001-01-01

288

Prevalence of a virus similar to human hepatitis B virus in swine  

Microsoft Academic Search

BACKGROUND: The objective of this study is to established evidence of the existence of a novel member of the hepadnavirus family endemic in swine. Temporarily this virus was designated as swine hepatitis B virus (SHBV). This SHBV can be detected by using human hepatitis B virus diagnostic kits including ELISA, immunohistochemical staining, and transmission electron microscopy (TEM). Also seroprevalence of

Wengui Li; Ruiping She; Liqiang Liu; Hua You; Jun Yin

2010-01-01

289

DNA modication and functional delivery into human cells using Escherichia coli DH10B  

E-print Network

DNA modi®cation and functional delivery into human cells using Escherichia coli DH10B Kumaran report a novel comprehen- sive Escherichia coli-based vector system for the modi®cation, propagation in recombination- de®cient Escherichia coli DH10B imparts greater stability to the large BAC clones but has made

Warburton, Peter E.

290

Genetic analysis of immortalizing functions of Epstein-Barr virus in human B lymphocytes  

Microsoft Academic Search

EPSTEIN-BARR virus (EBV), a herpes virus, infects human B lymphocytes in vitro and efficiently immortalizes them1. About 10 of the approximately 100 genes of EBV are expressed in recently immortalized B cells and although there is circumstantial evidence that at least three of these may contribute to the process of immortalization, there is no direct evidence that any particular gene

Wolfgang Hammerschmidt; Bill Sugden

1989-01-01

291

Human immunodeficiency virus, hepatitis B, C and D in Bangladesh's trucking industry: prevalence and risk factors  

Microsoft Academic Search

Background Human immunodeficiency virus (HIV) and hepatitis B and C, viral infections with shared percutaneous, mucosal and perinatal routes of transmission, are responsible for serious morbidity and mortality globally. In Bangladesh there is a dearth of research on prevalence and risk factors for these diseases. This study examines the prevalence of HIV and hepatitis (B, C, D) and risk factors

Laura Gibney; Parwez Choudhury

2001-01-01

292

Nicotine glucuronidation and the human UDP-glucuronosyltransferase UGT2B10.  

PubMed

Nicotine biotransformation affects the smoking habits of addicted individuals and therefore their health risk. Using an improved analytical method, we have discovered that the human UDP-glucuronosyltransferase (UGT) 2B10, a liver enzyme previously unknown to conjugate nicotine or exhibit considerable activity toward any compound, plays a major role in nicotine inactivation by direct conjugation with glucuronic acid at the aromatic nitrogen atom. The K(m) value of recombinant UGT2B10 for nicotine (0.29 mM) was similar to that determined for human liver microsomes (0.33 mM), whereas the K(m) value of UGT1A4 for nicotine was almost 10-fold greater (2.4 mM). UGT2B10 was also more active than UGT1A4 in N-glucuronidation of cotinine (oxidative nicotine metabolite), whereas UGT2B7 exhibited only low nicotine glucuronidation activity and was essentially inactive toward cotinine. UGT1A9 did not glucuronidate nicotine or cotinine. Quantitative reverse transcription-polymerase chain reaction showed that UGT2B10 mRNA was exclusively expressed in human liver, whereas UGTs 1A4 and 2B7 were expressed at comparable, although somewhat lower, levels in liver and several other extrahepatic tissues, including kidney and intestine. These findings for UGT2B10 (but not for UGT1A4 and UGT2B7) were mirrored by human tissue activities because nicotine and cotinine glucuronidation rates in intestine microsomes were less than 0.1% that of human liver microsomes. These novel findings solve two seemingly separate questions: which UGT is primarily responsible for nicotine glucuronidation in human liver, and what conjugation reactions are catalyzed by UGT2B10. PMID:17576790

Kaivosaari, Sanna; Toivonen, Pivi; Hesse, Leah M; Koskinen, Mikko; Court, Michael H; Finel, Moshe

2007-09-01

293

Human mast cell migration in response to members of the transforming growth factor-b family  

Microsoft Academic Search

Mast cells are known to accumulate at sites of inflammation, however, the chemotaxins involved remain largely undefined. Transforming growth factor-b (TGF-b) isoforms regulate numer- ous cellular functions, including cell growth and differentiation, formation of extracellular matrix, and the immune response. In this study we have compared the potency of different members of the TGF-b family as human mast cell chemotaxins,

Niclas Olsson; Ester Piek; Gunnar Nilsson

294

Activation of Epstein-Barr virus latent genes protects human B cells from death by apoptosis  

Microsoft Academic Search

EPSTEiN-Barr virus (EBV), a human herpesvirus, establishes a persistent asymptomatic infection of the circulating B-lymphocyte pool1-3. The mechanism of virus persistence is not understood but, given the limited lifespan of most B cells in vivo, it seems most likely that EBV-infected cells must gain access to the long-lived memory B-cell pool. Here we show in an in vitro system that

Christopher D. Gregory; Caroline Dive; Sheila Henderson; Christopher A. Smith; Gwyn T. Williams; John Gordon; Alan B. Rickinson

1991-01-01

295

Frequent Occult Hepatitis B Virus Infection in Patients Infected with Human Immunodeficiency Virus Type 1  

Microsoft Academic Search

\\u000a The presence of hepatitis B virus (HBV) serological markers was investigated in 170 patients (137 male, 33 female) infected\\u000a with human immunodeficiency virus (HIV) type 1. Antibodies to the hepatitis B core antigen (anti-HBc antibodies) were detected\\u000a in 115 (68%) patients. Of these 115, 14 (12%) were hepatitis B surface antigen (HBsAg) positive, 60 (52%) presented anti-HBs\\u000a antibodies, and

E. A. Santos; C. F. T. Yoshida; V. C. Rolla; J. M. Mendes; I. F. Vieira; J. Arabe; S. A. Gomes

2003-01-01

296

A complex selection signature at the human AVPR1B gene  

Microsoft Academic Search

BACKGROUND: The vasopressin receptor type 1b (AVPR1B) is mainly expressed by pituitary corticotropes and it mediates the stimulatory effects of AVP on ACTH release; common AVPR1B haplotypes have been involved in mood and anxiety disorders in humans, while rodents lacking a functional receptor gene display behavioral defects and altered stress responses. RESULTS: Here we have analyzed the two exons of

Rachele Cagliani; Matteo Fumagalli; Uberto Pozzoli; Stefania Riva; Matteo Cereda; Giacomo P Comi; Linda Pattini; Nereo Bresolin; Manuela Sironi

2009-01-01

297

NF-kappaB blockade and oncogenic Ras trigger invasive human epidermal neoplasia  

Microsoft Academic Search

The nuclear factor NF-kappaB and oncogenic Ras can alter proliferation in epidermis, the most common site of human cancer. These proteins are implicated in epidermal squamous cell carcinoma in mice, however, the potential effects of altering their function are uncertain. Whereas inhibition of NF-kappaB enhances apoptosis in certain tumours, blockade of NF-kappaB predisposes murine skin to squamous cell carcinoma. Because

Maya Dajee; Mirella Lazarov; Jennifer Y. Zhang; Ti Cai; Cheryl L. Green; Alan J. Russell; M. Peter Marinkovich; Shiying Tao; Qun Lin; Yoshiaki Kubo; Paul A. Khavari

2003-01-01

298

Clarithromycin Inhibits NFB Activation in Human Peripheral Blood Mononuclear Cells and Pulmonary Epithelial Cells  

Microsoft Academic Search

Macrolide antibiotics modulate the production of proinflammatory cytokines in vivo and in vitro. Tran- scription of the genes for these proinflammatory cytokines is regulated by nuclear factor kB (NF-kB). We ex- amined whether or not clarithromycin inhibits the activation of NF-kB induced by tumor necrosis factor alpha (TNF-a) or staphylococcal enterotoxin A (SEA) in human monocytic U-937 cells, a T-cell

TAKASHI ICHIYAMA; MIKI NISHIKAWA; TOMOMI YOSHITOMI; SHUNJI HASEGAWA; TOMOYO MATSUBARA; TAKASHI HAYASHI; SUSUMU FURUKAWA

2001-01-01

299

HUMAN B LYMPHOCYTE SUBSETS I. IgG-bearing B Cell Response to Pokeweed Mitogen  

Microsoft Academic Search

It is well established that B lymphocytes recognize the antigen to which they can respond by using antibody specific for that antigen as a surface receptor. In recent years it has become clear that most B cells display more than one class of immuno- globulin on their surface, although each of the two or three such molecules expressed have the

KENNETH A. AULT; MURRAY TOWLE

300

Reduction of tumor necrosis factor-alpha (TNF-?) related nuclear factor-kappaB (NF-?B) translocation but not inhibitor kappa-B (I?-B)-degradation by Rho protein inhibition in human endothelial cells  

Microsoft Academic Search

Degradation of inhibitor kappa-B (I?-B) followed by translocation of nuclear factor-kappaB (NF-?B) into the nucleus and activation of gene expression is essential in tumor necrosis factor-alpha (TNF-?)-signaling. In order to analyze the role of Rho proteins in TNF-?-induced NF-?B-activation in human umbilical cord vein endothelial cells (HUVEC) we used Clostridium difficile toxin B-10463 (TcdB-10463) which inactivates RhoA\\/Rac1\\/Cdc42 by glucosylation and

Stefan Hippenstiel; Bernd Schmeck; Joachim Seybold; Matthias Krll; Christoph v Eichel-Streiber; Norbert Suttorp

2002-01-01

301

Rapid Detection of Enteroviruses in Small Volumes of Natural Waters by Real-Time Quantitative Reverse Transcriptase PCR  

Microsoft Academic Search

Despite viral contamination of recreational waters, only bacterial, not viral, indicators are monitored routinely, due to a lack of rapid and cost-effective assays. We used negatively charged filters to capture enteroviruses from seawater and freshwater. Viral RNA was extracted using a commercial kit, and the viruses were quantified by real-time quantitative reverse transcriptase PCR (qRT-PCR). Poliovirus (6.6 to 330,000 virus

Jed A. Fuhrman; Xiaolin Liang; Rachel T. Noble

2005-01-01

302

Growth Inhibitory Effects of the Dual ErbB1\\/ErbB2 Tyrosine Kinase Inhibitor PKI166 on Human Prostate Cancer Xenografts1  

Microsoft Academic Search

Experiments with human prostate cancer cell lines have shown that forced overexpression of the ErbB2-receptor tyrosine kinase (RTK) pro- motes androgen-independent growth and increases androgen receptor- transcriptional activity in a ligand-independent fashion. To investigate the relationship between ErbB-RTK signaling and androgen in genetically unmanipulated human prostate cancer, we performed biochemical and biological studies with the dual ErbB1\\/ErbB2 RTK inhibitor PKI-166

Ingo K. Mellinghoff; Chris Tran; Charles L. Sawyers

2002-01-01

303

Comparison of human B cell activation by TLR7 and TLR9 agonists  

PubMed Central

Background Human B cells and plasmacytoid dendritic cells (pDC) are the only cells known to express both TLR7 and TLR9. Plasmacytoid dendritic cells are the primary IFN-? producing cells in response to TLR7 and TLR9 agonists. The direct effects of TLR7 stimulation on human B cells is less understood. The objective of this study was to compare the effects of TLR7 and TLR9 stimulation on human B cell function. Results Gene expression and protein production of cytokines, chemokines, various B cell activation markers, and immunoglobulins were evaluated. Purified human CD19+ B cells (99.9%, containing both nave and memory populations) from peripheral blood were stimulated with a TLR7-selective agonist (852A), TLR7/8 agonist (3M-003), or TLR9 selective agonist CpG ODN (CpG2006). TLR7 and TLR9 agonists similarly modulated the expression of cytokine and chemokine genes (IL-6, MIP1 alpha, MIP1 beta, TNF alpha and LTA), co-stimulatory molecules (CD80, CD40 and CD58), Fc receptors (CD23, CD32), anti-apoptotic genes (BCL2L1), certain transcription factors (MYC, TCFL5), and genes critical for B cell proliferation and differentiation (CD72, IL21R). Both agonists also induced protein expression of the above cytokines and chemokines. Additionally, TLR7 and TLR9 agonists induced the production of IgM and IgG. A TLR8-selective agonist was comparatively ineffective at stimulating purified human B cells. Conclusion These results demonstrate that despite their molecular differences, the TLR7 and TLR9 agonists induce similar genes and proteins in purified human B cells. PMID:18652679

Hanten, John A; Vasilakos, John P; Riter, Christie L; Neys, Lori; Lipson, Kenneth E; Alkan, Sefik S; Birmachu, Woubalem

2008-01-01

304

The influence of HLA B27 and interferon-? on the invasion and persistence of yersinia in primary human fibroblasts  

Microsoft Academic Search

To examine the role of the human HLA class I molecule B27 in the direct interaction of yersinia and human cells we performed\\u000a experimental infections of constitutive HLA B27-positive and -negative human fibroblast monolayers with Yersinia enterocolitica O.3. The numbers of yersiniae invading HLA B27-positive cells were within the range of HLA B27-negative cells. Treatment\\u000a of fibroblasts with interferon-? (IFN-?)

Hans-Iko Huppertz; Jrgen Heesemann

1996-01-01

305

Identification of a Novel Enterovirus E Isolates HY12 from Cattle with Severe Respiratory and Enteric Diseases  

PubMed Central

In this study, a virus strain designated as HY12 was isolated from cattle with a disease of high morbidity and mortality in Jilin province. Biological and physiochemical properties showed that HY12 isolates is cytopathic with an extremely high infectivity. HY12 is resistant to treatment of organic solvent and acid, and unstable at 60C for 1 h. Electron microscopy observation revealed the virus is an approximately 2228 nm in diameter. The complete genome sequence of HY12 consists of 7416 nucleotides, with a typical picornavirus genome organization including a 5?-untranslated region (UTR), a large single ORF encoding a polyprotein of 2176 amino acids, and a 3?-UTR. Phylogenetic analysis clustered HY12 isolates to a new serotype/genotype within the clade of enterovirus E (formerly BEV-A). Alignment analysis revealed a unique insertion of 2 amino acid residues (NF) at the C-terminal of VP1 protein between aa 825 and 826, and several rare mutations in VP1 and VP4 of HY12 isolates in relation to known bovine enterovirus (BEV) strains. This is the first report of an enterovirus E in China, which is potentially associated with an outbreak in cattle with severe respiratory and enteric diseases. PMID:24830424

Li, Sujing; San, Zhihao; Wang, Xinping

2014-01-01

306

Human apolipoprotein B RNA editing deaminase gene (APOBEC1).  

PubMed

Genomic clones encoding the human APOBEC1 gene and its 5' flanking region have been isolated and characterized. The human gene contains five coding exons. The introns dividing these exons correspond exactly to those found in the mouse gene. The translation initiation site, ATG, is located in exon 2 at the same site as in the mouse. The 5' flanking sequence contains two Alu repeats of the Sq family. Primer extension analysis demonstrated the presence of two major transcription initiation sites. The first transcription initiation site delineates the beginning of a noncoding first exon and resides downstream of the first Alu sequence. The second transcription initiation site is within the second Alu repeat. This Alu repeat resides within the first intron, which is spliced out of the transcript from the first start site. Neither transcription initiation site has a TATA or CCAT box. Comparison with the mouse gene suggests that the Alu sequence insertion split the intestinal promoter and that subsequently the down-stream Alu sequence took on a promoter function. No evidence was found for a far upstream non-tissue-specific promoter similar to that demonstrated in the mouse gene. Rather, consideration of results from the marsupial APOBEC-1 gene suggests that this upstream mouse promoter may have had a later evolutionary origin. PMID:9479499

Fujino, T; Navaratnam, N; Scott, J

1998-01-15

307

Coxsackievirus B detection in cases of myocarditis, myopericarditis, pericarditis and dilated cardiomyopathy in hospitalized patients.  

PubMed

Coxsackieviruses B (CV?B) are known as the most common viral cause of human heart infections. The aim of the present study was to assess the potential role of CV?B in the etiology of infectious heart disease in hospitalized patients. The present study is based on blood, pericardial fluid and heart biopsies from 102 patients and 100 control subjects. All of the samples were examined for the detection of specific enteroviral genome using the reverse transcription polymerase chain reaction (RT?PCR) and sequence analysis. Immunohistochemical investigations for the detection of the enteroviral capsid protein, VP1, from the biopsies were performed. The samples were cultured on confluent KB monolayer cell line for possible virus isolation. The epidemiological data were also collected. CV?B was detected in 28 of the 102patients. The sequence analysis demonstrated that 27 strains were identical to CV?B3 and only one strain was identical to CV?B1. Furthermore, VP1 in the heart biopsies was detected in enterovirus?positive cases, as revealed by RT?PCR. Pericarditis infection was more frequent than myocarditis (P<0.05) or myopericarditis (P=0.05). The epidemiological data demonstrate that CV?B heart infections occur mainly during autumn and winter, and young male adults are more susceptible than adolescents or adults (P<0.5). The present findings demonstrate a higher prevalence of viral heart infections, suggesting that CV?B may significantly contribute to heart infections. PMID:25241846

Gaaloul, Imed; Riabi, Samira; Harrath, Rafik; Hunter, Timothy; Hamda, Khaldoun B; Ghzala, Assia B; Huber, Sally; Aouni, Mahjoub

2014-12-01

308

Human Interferon Alpha-2b: A Therapeutic Protein for Cancer Treatment  

PubMed Central

Human interferon alpha (hIFN?) is a wide biological activity cytokine that is used in hepatitis and cancer treatments. It regulates many genes that are involved in antiviral and antiproliferative activities. This mini review focuses on human interferon alpha-2b (hIFN?-2b) as therapeutic protein for cancer treatment. The review covers hIFN?-2b molecular characteristic and its molecular mechanism by Janus activated kinase/signal transducer activation of transcription (JAK-STAT) pathway. The JAK-STAT pathway regulates not only proteins involved in inhibition of proliferation but also apoptosis. As additional discussion of clinical applications, the use of recombinant hIFN?-2b (rhIFN?-2b) as therapeutic protein in several types of cancer is also explained. PMID:24741445

Asmana Ningrum, Ratih

2014-01-01

309

Design and validation of a real-time RT-PCR for the simultaneous detection of enteroviruses and parechoviruses in clinical samples.  

PubMed

Human enteroviruses (EVs) and parechoviruses (HPeVs) are important etiological agents causing infections such as meningitis, encephalitis and sepsis-like disease in neonates and young children. We have developed a real-time RT-PCR for simultaneous detection of EV and HPeV in clinical samples. Primers and probe sets were designed from the conserved 5'-noncoding region of the genomes. The sensitivity, specificity and reproducibility of the technique were measured using a set of 25 EV and 6 HPeV types. All EVs but no HPeVs were detected with the EV primers-probe set. The HPeV primers-probe set detected only the 6 HPeV types. The lower detection limit was found to be 4 and 40CCID50/ml for HPeV and EV respectively, demonstrating high sensitivity of the technique for both viruses. The threshold cycle values were highly reproducible on repeat testing of positive controls among assay runs. The assay was evaluated in 53 clinical samples of suspected meningitis, sepsis or febrile syndromes from children under 3 years. In 11 of these (21%) EVs were detected, while 4, i.e. 7.5%, were HPeV positive. Molecular typing was carried out for 73% of the viruses. In summary, the RT-PCR method developed demonstrated effectively both EV and HPeV detection, which can cause similar clinical symptoms in infants. PMID:25152526

Cabrerizo, Mara; Calvo, Cristina; Rabella, Nuria; Muoz-Almagro, Carmen; Del Amo, Eva; Prez-Ruiz, Mercedes; Sanbonmatsu-Gmez, Sara; Moreno-Docn, Antonio; Otero, Almudena; Trallero, Gloria

2014-11-01

310

Selective suppression of an early step in human B cell activation by cyclosporin A  

PubMed Central

The effect of cyclosporin A (CsA), a fungal metabolite with immunosuppressive properties, on the induction of human B cell proliferation and differentiation, has been described. CsA had a selective inhibitory effect on the activation phase of the cell cycle vs. the proliferation phase following preactivation of the cells. Cell enlargement and RNA synthesis of small resting B cells triggered by anti-mu were inhibited by addition of CsA (5-500 ng/ml). The inhibitory effect of CsA was found only when the drug was added within 24 h of initiation of culture. In marked contrast, once small B cells were activated by anti-mu, the resulting large, activated B cells could be induced to initiate DNA synthesis by incubation with B cell growth factor (BCGF), and addition of CsA (1-1,000 ng/ml) to the culture did not suppress this BCGF-induced B cell proliferation. Addition of CsA to cultures of B cells which had been preactivated with Staphylococcus aureus Cowan strain I (SAC) and were already proliferating did not suppress B cell differentiation factor (BCDF)-induced differentiation of these cells. Thus, these data indicate that CsA can be used as a pharmacologic tool to dissect out human B cell responses into two distinct steps: (a) the initial activation step induced by anti-Ig, which is characterized by cell enlargement, RNA synthesis, and expression of receptors for BCGF; and (b) the proliferative step induced by BCGF in these preactivated B cells that undergo DNA synthesis and can then go on to differentiate in the presence of BCDF. In this regard, CsA selectively suppresses an early step of human B cell activation and has little inhibitory effect on the subsequent factor-dependent proliferation and differentiation. PMID:6193229

1983-01-01

311

New Coxsackievirus B4 Genotype Circulating in Inner Mongolia Autonomous Region, China  

PubMed Central

Hand, foot, and mouth disease (HFMD) surveillance was initiated in the Inner Mongolia Autonomous Region of China in 2007, a crucial scrutiny for monitoring the prevalence of enterovirus serotypes associated with HFMD patients. However, this surveillance mostly focused on enterovirus 71 (EV-A71) and coxsackievirus A16; therefore, information on other enterovirus serotypes is limited. To identify the other circulating enterovirus serotypes in the HFMD outbreaks in Inner Mongolia in 2010, clinical samples from HFMD patients were investigated. Six coxsackievirus B4 (CVB4) strains were isolated and phylogenetic analyses of VP1 sequences were performed. Full-length genome sequences of two representative CVB4 isolates were acquired and similarity plot and bootscanning analyses were performed. The phylogenetic dendrogram indicated that all CVB4 strains could be divided into 5 genotypes (Genotypes IV) with high bootstrap support (90100%). The CVB4 prototype strain (JVB) was the sole member of genotype I. CVB4 strains belonging to genotype II, which were once common in Europe and the Americas, seemingly disappeared and gave way to genotype III and IV strains, which appear to be the dominant circulating strains in the world. All Chinese CVB4 strains belonged to Genotype V, a newly identified genotype supported by a high bootstrap value (100%), and are circulating only in mainland of China. Intertypic recombination occurred in the Chinese CVB4 strains with novel unknown serotype EV-B donor sequences. Two Chinese CVB4 strains had a virulent residue at position 129 of VP1, and one strain also had a virulent residue at position 16 of VP4. Increased surveillance is needed to monitor the emergence of new genetic lineages of enteroviruses in areas that are often associated with large-scale outbreaks. In addition, continued monitoring of enteroviruses by clinical surveillance and genetic characterization should be enhanced. PMID:24595311

Gu, Suyi; Fan, Yaochun; Sun, Qiang; Zhang, Bo; Yan, Shaohong; Xu, Wenbo; Ma, Xueen; Wang, Wenrui

2014-01-01

312

Structures of human monoamine oxidase B complexes with selective noncovalent inhibitors: safinamide and coumarin analogs.  

PubMed

Structures of human monoamine oxidase B (MAO B) in complex with safinamide and two coumarin derivatives, all sharing a common benzyloxy substituent, were determined by X-ray crystallography. These compounds competitively inhibit MAO B with Ki values in the 0.1-0.5 microM range that are 30-700-fold lower than those observed with MAO A. The inhibitors bind noncovalently to MAO B, occupying both the entrance and the substrate cavities and showing a similarly oriented benzyloxy substituent. PMID:17915852

Binda, Claudia; Wang, Jin; Pisani, Leonardo; Caccia, Carla; Carotti, Angelo; Salvati, Patricia; Edmondson, Dale E; Mattevi, Andrea

2007-11-15

313

Human and mouse homologs of Escherichia coli DinB (DNA polymerase IV), members of the UmuC/DinB superfamily  

PubMed Central

To understand the mechanisms underlying mutagenesis in eukaryotes better, we have cloned mouse and human homologs of the Escherichia coli dinB gene. E. coli dinB encodes DNA polymerase IV and greatly increases spontaneous mutations when overexpressed. The mouse and human DinB1 amino acid sequences share significant identity with E. coli DinB, including distinct motifs implicated in catalysis, suggesting conservation of the polymerase function. These proteins are members of a large superfamily of DNA damage-bypass replication proteins, including the E. coli proteins UmuC and DinB and the Saccharomyces cerevisiae proteins Rev1 and Rad30. In a phylogenetic tree, the mouse and human DinB1 proteins specifically group with E. coli DinB, suggesting a mitochondrial origin for these genes. The human DINB1 gene is localized to chromosome 5q13 and is widely expressed. PMID:10518552

Gerlach, Valerie L.; Aravind, L.; Gotway, Garrett; Schultz, Roger A.; Koonin, Eugene V.; Friedberg, Errol C.

1999-01-01

314

Control of the Human B-Globin Gene  

NSDL National Science Digital Library

The diagram shows some of the gene regulatory proteins thought to control expression of this gene during red blood cell development. Some of the gene regulatory proteins shown, such as CP1, are found in many types of cells, while others, such as GATA-1, are present in only a few types of cells, including red blood cell precursors, and are therefore thought to contribute to the cell-type specificity of beta-globin gene expression. As indicated by the bidirectional arrows, several of the binding sites for GATA-1 overlap those of other gene regulatory proteins; it is thought that occupancy of these sites by GATA-1 excludes binding of other proteins. (Adapted from B. Emerson, In Gene Expression: General and Cell-Type Specific (M. Karin, ed.), pp. 116-161. Boston: Birkhauser, 1993.)

BEGIN:VCARD VERSION:2.1 FN:Bruce Alberts N:Alberts;Bruce REV:2005-04-16 END:VCARD; BEGIN:VCARD VERSION:2.1 FN:Dennis Bray N:Bray;Dennis REV:2005-04-16 END:VCARD; BEGIN:VCARD VERSION:2.1 REV:2005-04-16 END:VCARD; BEGIN:VCARD VERSION:2.1 FN:Alexander Johnson N:Johnson;Alexander REV:2005-04-16 END:VCARD; BEGIN:VCARD VERSION:2.1 FN:Julian Lewis N:Lewis;Julian REV:2005-04-16 END:VCARD; BEGIN:VCARD VERSION:2.1 FN:Martin Raff N:Raff;Martin REV:2005-04-16 END:VCARD

1998-07-01

315

Expression of human {beta}-defensin-2 gene induced by CpG-DNA in human B cells  

SciTech Connect

Defensins have a broad range of antimicrobial activity against bacteria, fungi, and viruses. The expression of human {beta}-defensin-2 (hBD-2) is prevalently observed in epithelial cells and is induced by bacterial infection. Here, we have shown that the expression of the hBD-2 gene and release of hBD-2 protein into the medium is up-regulated in response to CpG-DNA in human B cell line RPMI 8226. The induction of hBD-2 was dependent on CG sequence and phosphorothioate backbone-modification. This was also confirmed in primary human lymphocytes. To shed light on the molecular mechanism involved in hBD-2 induction by CpG-DNA, we examined the contribution of the NF-{kappa}B signaling pathway in RPMI 8226 cells. Suppression of MyD88 function and inhibition of NF-{kappa}B nuclear localization blocked hBD-2 induction. The NF-{kappa}B pathway inhibitors also abolished hBD-2 induction. These results may contribute to a better understanding on the therapeutic effects of CpG-DNA against infectious diseases.

Han, Su Ho [Department of Microbiology, College of Medicine, Hallym University, Gangwon-do 200-702 (Korea, Republic of)] [Department of Microbiology, College of Medicine, Hallym University, Gangwon-do 200-702 (Korea, Republic of); Kim, Young-Eun; Park, Jeong-A [Department of Biochemistry, College of Natural Sciences, Chungbuk National University, Chungbuk 361-763 (Korea, Republic of)] [Department of Biochemistry, College of Natural Sciences, Chungbuk National University, Chungbuk 361-763 (Korea, Republic of); Park, Jae-Bong [Department of Biochemistry, College of Medicine, Hallym University, Gangwon-do 200-702 (Korea, Republic of)] [Department of Biochemistry, College of Medicine, Hallym University, Gangwon-do 200-702 (Korea, Republic of); Kim, Yong-Sun [Department of Microbiology, College of Medicine, Hallym University, Gangwon-do 200-702 (Korea, Republic of)] [Department of Microbiology, College of Medicine, Hallym University, Gangwon-do 200-702 (Korea, Republic of); Lee, Younghee [Department of Biochemistry, College of Natural Sciences, Chungbuk National University, Chungbuk 361-763 (Korea, Republic of)] [Department of Biochemistry, College of Natural Sciences, Chungbuk National University, Chungbuk 361-763 (Korea, Republic of); Choi, Ihn-Geun [Department of Neuropsychiatry, Hallym University, Han-Gang Sacred Heart Hospital, Seoul 150-719 (Korea, Republic of)] [Department of Neuropsychiatry, Hallym University, Han-Gang Sacred Heart Hospital, Seoul 150-719 (Korea, Republic of); Kwon, Hyung-Joo, E-mail: hjookwon@hallym.ac.kr [Department of Microbiology, College of Medicine, Hallym University, Gangwon-do 200-702 (Korea, Republic of) [Department of Microbiology, College of Medicine, Hallym University, Gangwon-do 200-702 (Korea, Republic of); Center for Medical Science Research, College of Medicine, Hallym University, Gangwon-do 200-702 (Korea, Republic of)

2009-11-20

316

Scored Antibody Reactivity Determined by Immunoblotting Shows an Association between Clinical Manifestations and Presence of Borrelia burgdorferi sensu stricto, B. garinii, B. afzelii, and B. Valaisiana in Humans  

PubMed Central

An immunoglobulin G immunoblot was developed with antigenic extracts of Borrelia burgdorferi sensu stricto, B. garinii, B. afzelii, and B. valaisiana genospecies and was reacted with sera from patients with neuroborreliosis, acrodermatitis, and Lyme arthritis. A detailed analysis of the reactivities of the protein bands was performed, and a two-step scoring procedure was selected to determine the preferential reactivity of sera to one particular genospecies. The discriminative potential of 5 proteins (12-kDa, 16-kDa, 18-kDa, OspA, and 66-kDa proteins) was used as a rapid first-step scoring method, followed by scoring of 14 additional protein bands if necessary. The advantage of this procedure is the low percentage of serum samples with inconclusive results for one of the four species (10% for patients with neuroborreliosis, 6% for patients with acrodermatitis chronica atrophicans, and 6% for patients with Lyme arthritis). Among 31 serum samples from patients with neuroborreliosis, 16 were more reactive to B. garinii, 7 were more reactive to B. afzelii, 3 were more reactive to B. valaisiana, and 2 were more reactive to B. burgdorferi sensu stricto. Of 31 serum samples from patients with acrodermatitis, 26 showed a higher level of reactivity to B. afzelii. Of 34 serum samples from patients with Lyme arthritis, 21 were more reactive to B. burgdorferi sensu stricto, 10 were more reactive to B. afzelii, and 1 was more reactive to B. valaisiana. Our results suggest an organotropism of Borrelia species and provide some evidence of a pathogenic potential of B. valaisiana in humans. PMID:10565936

Ryffel, Karine; Peter, Olivier; Rutti, Bernard; Suard, Andre; Dayer, Eric

1999-01-01

317

Membrane-associated signaling in human B-lymphoma lines  

SciTech Connect

In B-non-Hodgkin lymphomas, Lyn and Cbp/PAG constitute the core of an oncogenic signalosome that captures the Phosphatidylinositol-3-kinase, the Spleen tyrosine kinase and the Signal transducer and activator of transcription-3 to generate pro-survival and proliferative signals. Lymphoma lines corresponding to follicular, mantle-cell and Burkitt-derived lymphomas display type-specific signalosome organizations that differentially activate PI3K, Syk and STAT3. In the follicular lymphoma line, PI3K, Syk and STAT3 were optimally activated upon association with the Lyn-Cbp/PAG signalosome, while in the Burkitt lymphoma-derived line, the association with Cbp/PAG and activation of PI3K were interfered with by the latent membrane proteins encoded by the Epstein-Barr virus. In the Jeko-1 mantle-cell line, a weak association of Syk with the Lyn-Cbp/PAG signalosome resulted in poor activation of Syk, but in those cells, as in the follicular and Burkitt-derived lines, efficient apoptosis induction by the Syk inhibitor R406 indicated that Syk is nonetheless an important prosurvival element and therefore a valuable therapeutic target. In all configurations described herein is the Lyn-Cbp/PAG signalosome independent of external signals and provides efficient means of activation for its associated lipid and protein kinases. In follicular and Burkitt-derived lines, Syk appears to be activated following binding to Cbp/PAG and no longer requires B-cell receptor-associated activation motifs for activation. Assessment of the different modalities of Lyn-Cbp/PAG signalosome organization could help in selecting the appropriate combination of kinase inhibitors to eliminate a particular type of lymphoma cells.

Tauzin, Sebastien; Ding, Heidrun; Burdevet, Dimitri [Department of Pathology and Immunology, Centre medical universitaire, 1, rue Michel-Servet, 1211 Geneva 11 (Switzerland)] [Department of Pathology and Immunology, Centre medical universitaire, 1, rue Michel-Servet, 1211 Geneva 11 (Switzerland); Borisch, Bettina [Department of Social and Preventive Medicine, Centre medical universitaire, 1, rue Michel-Servet, 1211 Geneva 11 (Switzerland)] [Department of Social and Preventive Medicine, Centre medical universitaire, 1, rue Michel-Servet, 1211 Geneva 11 (Switzerland); Hoessli, Daniel C., E-mail: danielhoessli@gmail.com [Department of Pathology and Immunology, Centre medical universitaire, 1, rue Michel-Servet, 1211 Geneva 11 (Switzerland)

2011-01-15

318

Murine and human b locus pigmentation genes encode a glycoprotein (gp75) with catalase activity  

SciTech Connect

Melanogenesis is regulated in large part by tyrosinase, and defective tyrosinase leads to albinism. The mechanisms for other pigmentation determinants (e.g., those operative in tyrosinase-positive albinism and in murine coat-color mutants) are not yet known. One murine pigmentation gene, the brown (b) locus, when mutated leads to a brown (b/b) or hypopigmentated (B{sup lt}/B{sup lt}) coat versus the wild-type black (B/B). The authors show that the b locus codes for a glycoprotein with the activity of a catalase (catalase B). Only the c locus protein is a tyrosinase. Because peroxides may be by-products of melanogenic activity and hydrogen peroxide in particular is known to destroy melanin precursors and melanin, they conclude that pigmentation is controlled not only by tyrosinase but also by a hydroperoxidase. The studies indicate that catalase B is identical with gp75, a known human melanosomal glycoprotein; that the b mutation is in a heme-associated domain; and that the B{sup lt} mutation renders the protein susceptible to rapid proteolytic degradation.

Halaban, R.; Moellmann, G. (Yale Univ. School of Medicine, New Haven, CT (USA))

1990-06-01

319

Inhibitory effects of acyclic nucleoside phosphonates on human hepatitis B virus and duck hepatitis B virus infections in tissue culture.  

PubMed Central

The inhibitory effects of the 9-(2-phosphonylmethoxyethyl)adenine-related compounds (S)-9-(3-hydroxy-2-phosphonylmethoxypropyl)-adenine, (S)-9-(3-fluoro-2-phosphonylmethoxypropyl)adenine, (R)-9-(2-phosphonylmethoxypropyl)adenine, (R)-9-(2-phosphonylmethoxypropyl)-2,6-diaminopurine, and (S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine on human hepatitis B virus replication in the human hepatoma cell line HepG2 2.2.15 and duck hepatitis B virus infection in primary duck hepatocytes were investigated. (R)-9-(2-phosphonylmethoxypropyl-2,6-diaminopurine had the lowest 50% inhibitory concentrations against hepatitis B virus and duck hepatitis B virus, 0.22 and 0.06 microM, respectively, i.e., two- to fivefold lower concentrations than required for (R)-9-(2-phosphonylmethoxypropyl)adenine and 9-(2-phosphonylmethoxyethyl)adenine. All compounds were not toxic in vitro at a concentration of 100 microM. PMID:7811041

Heijtink, R A; Kruining, J; de Wilde, G A; Balzarini, J; de Clercq, E; Schalm, S W

1994-01-01

320

Corruption of human follicular B-lymphocyte trafficking by a B-cell superantigen.  

PubMed

Protein A (SpA) of Staphylococcus aureus is known to target the paratope of immunoglobulins expressing V(H)3 genes, and to delete marginal zone B cells and B-1a in vivo. We have discovered that SpA endows S. aureus with the potential to subvert B-cell trafficking in the host. We found that SpA, whose Fc-binding site has been inactivated, binds essentially to nave B cells and induces a long-lasting decrease in CXCR4 expression and in B-cell chemotaxis to CXCL12. Competition experiments indicated that SpA does not interfere with binding of CXCR4 ligands and does not directly bind to CXCR4. This conclusion is strongly supported by the inability of SpA to modulate clathrin-mediated CXCR4 internalization, which contrasts with the potent effect of anti-immunoglobin M (IgM) antibodies. Microscopy and biochemical experiments confirmed that SpA binds to the surface IgM/IgD complex and induces its clathrin-dependent internalization. Concomitantly, the SpA-induced signaling leads to protein kinase C-dependent CXCR4 downmodulation, suggesting that SpA impairs the recycling of CXCR4, a postclathrin process that leads to either degradation into lysozomes or de novo expression at the cell surface. In addition to providing novel insight into disruption of B-cell trafficking by an infectious agent, our findings may have therapeutic implications. Because CXCR4 has been associated with cancer metastasis and with certain autoimmune diseases, SpA behaves as an evolutionary tailored highly specific, chemokine receptor inhibitor that may have value in addition to conventional cytotoxic therapy in patients with various malignancies and immune-mediated diseases. PMID:22367177

Borhis, Gwenoline; Viau, Muriel; Badr, Gamal; Richard, Yolande; Zouali, Moncef

2012-01-01

321

Corruption of Human Follicular B-Lymphocyte Trafficking by a B-Cell Superantigen  

PubMed Central

Protein A (SpA) of Staphylococcus aureus is known to target the paratope of immunoglobulins expressing VH3 genes, and to delete marginal zone B cells and B-1a in vivo. We have discovered that SpA endows S. aureus with the potential to subvert B-cell trafficking in the host. We found that SpA, whose Fc-binding site has been inactivated, binds essentially to nave B cells and induces a long-lasting decrease in CXCR4 expression and in B-cell chemotaxis to CXCL12. Competition experiments indicated that SpA does not interfere with binding of CXCR4 ligands and does not directly bind to CXCR4. This conclusion is strongly supported by the inability of SpA to modulate clathrin-mediated CXCR4 internalization, which contrasts with the potent effect of anti-immunoglobin M (IgM) antibodies. Microscopy and biochemical experiments confirmed that SpA binds to the surface IgM/IgD complex and induces its clathrin-dependent internalization. Concomitantly, the SpA-induced signaling leads to protein kinase Cdependent CXCR4 downmodulation, suggesting that SpA impairs the recycling of CXCR4, a postclathrin process that leads to either degradation into lysozomes or de novo expression at the cell surface. In addition to providing novel insight into disruption of B-cell trafficking by an infectious agent, our findings may have therapeutic implications. Because CXCR4 has been associated with cancer metastasis and with certain autoimmune diseases, SpA behaves as an evolutionary tailored highly specific, chemokine receptor inhibitor that may have value in addition to conventional cytotoxic therapy in patients with various malignancies and immune-mediated diseases. PMID:22367177

Borhis, Gwenoline; Viau, Muriel; Badr, Gamal; Richard, Yolande; Zouali, Moncef

2012-01-01

322

Expression and function of IL-12 and IL-18 receptors on human tonsillar B cells.  

PubMed

IL-12 activates murine and human B cells, but little information is available as to the expression and function of IL-12R on human B lymphocytes. Here we show that the latter cells, freshly isolated from human tonsils, expressed the transcripts of both beta1 and beta2 chains of IL-12R and that beta2 chain mRNA was selectively increased (4- to 5-fold) by incubation with Staphylococcus aureus Cowan I bacteria or IL-12. B cell stimulation with IL-12 induced de novo expression of the transcripts of the two chains of IL-18R, i.e., IL-1 receptor-related protein and accessory protein-like. Functional studies showed that both IL-12 and IL-18 signaled to B cells through the NF-kappaB pathway. In the case of IL-12, no involvement of STAT transcription factors, and in particular of STAT-4, was detected. c-rel and p50 were identified as the members of NF-kappaB family involved in IL-12-mediated signal transduction to B cells. IL-12 and IL-18 synergized in the induction of IFN-gamma production by tonsillar B cells, but not in the stimulation of B cell differentiation, although either cytokine promoted IgM secretion in culture supernatants. Finally, naive but not germinal center or memory, tonsillar B cells were identified as the exclusive IL-12 targets in terms of induction of NF-kappaB activation and of IFN-gamma production. PMID:11120812

Airoldi, I; Gri, G; Marshall, J D; Corcione, A; Facchetti, P; Guglielmino, R; Trinchieri, G; Pistoia, V

2000-12-15

323

Tumor necrosis factor alpha is an autocrine growth factor for normal human B cells.  

PubMed Central

Transcription of the human tumor necrosis factor alpha (TNF-alpha) gene is one of the earliest events that occurs after stimulation of B or T cells via their antigen receptors. Antibody directed at surface immunoglobulin (anti-Ig) on B cells has previously been shown to induce a rapid burst of TNF-alpha gene transcription, which can be blocked by the immunosuppressants cyclosporin A (CsA) and FK506. Here, TNF-alpha gene transcription is shown also to be highly and rapidly induced in human B cells after stimulation via the CD40 and interleukin 4 pathways, which similarly is inhibited by CsA and a panel of CsA or FK506 analogues that block calcineurin phosphatase activity. Endogenous TNF-alpha produced after stimulation was involved in B-cell proliferation since anti-TNF-alpha monoclonal antibody inhibited both anti-Ig- and anti-CD40-induced B-cell proliferative responses. Moreover, addition of TNF-alpha during stimulation resulted in augmentation of B-cell proliferation, which was also inhibited by anti-TNF-alpha monoclonal antibody. Although lymphotoxin alpha (LT-alpha) mRNA is induced by both pathways, it is not blocked by CsA, whereas LT-beta mRNA is constitutively expressed in B cells. Thus, TNF-alpha is a necessary autocrine growth factor for human B cells stimulated via two independent CsA-sensitive pathways and plays a role similar to that of interleukin 2 in T-cell proliferation. The autocrine nature of TNF-alpha in activated B cells implies a potential role for this cytokine in infection-related polyclonal B-cell expansion and in B-cell malignancies. Images PMID:7518925

Boussiotis, V A; Nadler, L M; Strominger, J L; Goldfeld, A E

1994-01-01

324

Granzyme B produced by human plasmacytoid dendritic cells suppresses T-cell expansion  

PubMed Central

Human plasmacytoid dendritic cells (pDCs) are crucially involved in the modulation of adaptive T-cell responses in the course of neoplastic, viral, and autoimmune disorders. In several of these diseases elevated extracellular levels of the serine protease granzyme B (GrB) are observed. Here we demonstrate that human pDCs can be an abundant source of GrB and that such GrB+ pDCs potently suppress T-cell proliferation in a GrB-dependent, perforin-independent manner, a process reminiscent of regulatory T cells. Moreover, we show that GrB expression is strictly regulated on a transcriptional level involving Janus kinase 1 (JAK1), signal transducer and activator of transcription 3 (STAT3), and STAT5 and that interleukin-3 (IL-3), a cytokine secreted by activated T cells, plays a central role for GrB induction. Moreover, we find that the immunosuppressive cytokine IL-10 enhances, while Toll-like receptor agonists and CD40 ligand strongly inhibit, GrB secretion by pDCs. GrB-secreting pDCs may play a regulatory role for immune evasion of tumors, antiviral immune responses, and autoimmune processes. Our results provide novel information about the complex network of pDCT-cell interactions and may contribute to an improvement of prophylactic and therapeutic vaccinations. PMID:19965634

Vollmer, Angelika; Blackwell, Sue E.; Maier, Julia; Sontheimer, Kai; Beyer, Thamara; Mandel, Birgit; Lunov, Oleg; Tron, Kyrylo; Nienhaus, G. Ulrich; Simmet, Thomas; Debatin, Klaus-Michael; Weiner, George J.

2010-01-01

325

Accumulation of peripheral autoreactive B cells in the absence of functional human regulatory T cells  

PubMed Central

Regulatory T cells (Tregs) play an essential role in preventing autoimmunity. Mutations in the forkhead box protein 3 (FOXP3) gene, which encodes a transcription factor critical for Treg function, result in a severe autoimmune disorder and the production of various autoantibodies in mice and in IPEX (immune dysregulation, polyendocrinopathy, enteropathy, X-linked) patients. However, it is unknown whether Tregs normally suppress autoreactive B cells. To investigate a role for Tregs in maintaining human B-cell tolerance, we tested the reactivity of recombinant antibodies isolated from single B cells isolated from IPEX patients. Characteristics and reactivity of antibodies expressed by new emigrant/transitional B cells from IPEX patients were similar to those from healthy donors, demonstrating that defective Treg function does not impact central B-cell tolerance. In contrast, mature naive B cells from IPEX patients often expressed autoreactive antibodies, suggesting an important role for Tregs in maintaining peripheral B-cell tolerance. T cells displayed an activated phenotype in IPEX patients, including their Treg-like cells, and showed up-regulation of CD40L, PD-1, and inducibl T-cell costimulator (ICOS), which may favor the accumulation of autoreactive mature naive B cells in these patients. Hence, our data demonstrate an essential role for Tregs in the establishment and the maintenance of peripheral B-cell tolerance in humans. PMID:23223361

Kinnunen, Tuure; Chamberlain, Nicolas; Morbach, Henner; Choi, Jinyoung; Kim, Sangtaek; Craft, Joseph; Mayer, Lloyd; Cancrini, Caterina; Passerini, Laura; Bacchetta, Rosa; Ochs, Hans D.; Torgerson, Troy R.

2013-01-01

326

Accumulation of peripheral autoreactive B cells in the absence of functional human regulatory T cells.  

PubMed

Regulatory T cells (Tregs) play an essential role in preventing autoimmunity. Mutations in the forkhead box protein 3 (FOXP3) gene, which encodes a transcription factor critical for Treg function, result in a severe autoimmune disorder and the production of various autoantibodies in mice and in IPEX (immune dysregulation, polyendocrinopathy, enteropathy, X-linked) patients. However, it is unknown whether Tregs normally suppress autoreactive B cells. To investigate a role for Tregs in maintaining human B-cell tolerance, we tested the reactivity of recombinant antibodies isolated from single B cells isolated from IPEX patients. Characteristics and reactivity of antibodies expressed by new emigrant/transitional B cells from IPEX patients were similar to those from healthy donors, demonstrating that defective Treg function does not impact central B-cell tolerance. In contrast, mature naive B cells from IPEX patients often expressed autoreactive antibodies, suggesting an important role for Tregs in maintaining peripheral B-cell tolerance. T cells displayed an activated phenotype in IPEX patients, including their Treg-like cells, and showed up-regulation of CD40L, PD-1, and inducibl T-cell costimulator (ICOS), which may favor the accumulation of autoreactive mature naive B cells in these patients. Hence, our data demonstrate an essential role for Tregs in the establishment and the maintenance of peripheral B-cell tolerance in humans. PMID:23223361

Kinnunen, Tuure; Chamberlain, Nicolas; Morbach, Henner; Choi, Jinyoung; Kim, Sangtaek; Craft, Joseph; Mayer, Lloyd; Cancrini, Caterina; Passerini, Laura; Bacchetta, Rosa; Ochs, Hans D; Torgerson, Troy R; Meffre, Eric

2013-02-28

327

Impaired T cell activation and cytokine production by calcitriol-primed human B cells.  

PubMed

The biologically active form of vitamin D3 , 1, 25-dihydroxyvitamin D3 (calcitriol), is a potent modulator of the immune response. We have shown previously that calcitriol modulates the immunoglobulin response in vitro and in vivo in mice and humans. To analyse the underlying molecular mechanisms we studied whether calcitriol-primed B cells modulate T cell activation and function. Human B cells were stimulated with anti-CD40 and interleukin (IL)-4 in the presence of increasing concentrations of calcitriol. After removal of calcitriol, primed B cells were co-cultured with autologous CD4(+) T cells; the B cell phenotype T cell activation and their consecutive cytokine production were also assessed. Naive T cells co-cultured with calcitriol-primed naive B cells showed a reduced expansion, nuclear factor of activated T cells, cytoplasmic 2 (NFATc2) expression and cytokine production upon restimulation. CD86 expression on B cells after calcitriol priming was identified as an underlying mechanism, as T cell activation and expansion was rescued by activating anti-CD28 antibodies. Our data indicate that calcitriol-primed B cells display an impaired capacity to activate T cells. Taken together, we identified a novel B cell-dependent vitamin D immune regulatory mechanism, namely by decreased co-stimulation of calcitriol-primed B cells. PMID:24965738

Drozdenko, G; Scheel, T; Heine, G; Baumgrass, R; Worm, M

2014-11-01

328

Effect of increased HoxB4 on human megakaryocytic development  

SciTech Connect

Research highlights: {yields} HoxB4 overexpression in human TF1 cells increased the expression of CD61 and CD41a. {yields} HoxB4 fusion protein enhanced megakaryocytic development of CD34{sup +} cord blood cells. {yields} Ectopic HoxB4 increased Tpo receptor expression and decreased c-Myb expression. {yields} HoxB4 RNA silencing increased c-Myb expression and decreased Fli-1 expression. -- Abstract: In order to produce clinically useful quantities of platelets ex vivo we may need to firstly enhance early self-renewal of hematopoietic stem cells (HSCs) and/or megakaryocyte (Mk) progenitors. The homeodomain transcription factor HoxB4 has been shown to be an important regulator of stem cell renewal and hematopoiesis; however, its effect on megakaryopoiesis is unclear. In this study, we investigated the effect of HoxB4 overexpression or RNA silencing on megakaryocytic development in the human TF1 progenitor cell line; we then used recombinant tPTD-HoxB4 fusion protein to study the effect of exogenous HoxB4 on megakaryocytic development of human CD34 positively-selected cord blood cells. We found that ectopic HoxB4 in TF1 cells increased the antigen expression of CD61and CD41a, increased the gene expression of thrombopoietin receptor (TpoR), Scl-1, Cyclin D1, Fog-1 and Fli-1 while it decreased c-Myb expression. HoxB4 RNA silencing in TF1 cells decreased the expression of CD61 and CD41a and decreased Fli-1 expression while it increased the expression of c-Myb. Recombinant tPTD-HoxB4 fusion protein increased the percentages and absolute numbers of CD41a and CD61 positive cells during megakaryocytic differentiation of CD34 positively-selected cord blood cells and increased the numbers of colony-forming unit-megakaryocyte (CFU-Mk). Adding tPTD-HoxB4 fusion protein increased the gene expression of TpoR, Cyclin D1, Fog-1 and Fli-1 while it inhibited c-Myb expression. Our data suggest that increased HoxB4 enhanced early megakaryocytic development in human TF1 cells and CD34 positively-selected cord blood cells primarily by upregulating TpoR and Fli-1 expression and downregulating c-Myb expression. Increasing HoxB4 expression or adding recombinant HoxB4 protein might be a way to expand Mks for the production of platelets for use in transfusion medicine.

Zhong, Yiming [Department of Pathology, The Ohio State University, Columbus, OH (United States) [Department of Pathology, The Ohio State University, Columbus, OH (United States); Program in Molecular, Cellular, and Developmental Biology, The Ohio State University, Columbus, OH (United States); Sullenbarger, Brent [Department of Pathology, The Ohio State University, Columbus, OH (United States)] [Department of Pathology, The Ohio State University, Columbus, OH (United States); Lasky, Larry C., E-mail: Lasky.4@osu.edu [Department of Pathology, The Ohio State University, Columbus, OH (United States); Program in Molecular, Cellular, and Developmental Biology, The Ohio State University, Columbus, OH (United States)

2010-07-30

329

Identification of STAT5A and STAT5B Target Genes in Human T Cells  

PubMed Central

Signal transducer and activator of transcription (STAT) comprises a family of universal transcription factors that help cells sense and respond to environmental signals. STAT5 refers to two highly related proteins, STAT5A and STAT5B, with critical function: their complete deficiency is lethal in mice; in humans, STAT5B deficiency alone leads to endocrine and immunological problems, while STAT5A deficiency has not been reported. STAT5A and STAT5B show peptide sequence similarities greater than 90%, but subtle structural differences suggest possible non-redundant roles in gene regulation. However, these roles remain unclear in humans. We applied chromatin immunoprecipitation followed by DNA sequencing using human CD4+ T cells to detect candidate genes regulated by STAT5A and/or STAT5B, and quantitative-PCR in STAT5A or STAT5B knock-down (KD) human CD4+ T cells to validate the findings. Our data show STAT5A and STAT5B play redundant roles in cell proliferation and apoptosis via SGK1 interaction. Interestingly, we found a novel, unique role for STAT5A in binding to genes involved in neural development and function (NDRG1, DNAJC6, and SSH2), while STAT5B appears to play a distinct role in T cell development and function via DOCK8, SNX9, FOXP3 and IL2RA binding. Our results also suggest that one or more co-activators for STAT5A and/or STAT5B may play important roles in establishing different binding abilities and gene regulation behaviors. The new identification of these genes regulated by STAT5A and/or STAT5B has major implications for understanding the pathophysiology of cancer progression, neural disorders, and immune abnormalities. PMID:24497979

Kanai, Takahiro; Seki, Scott; Jenks, Jennifer A.; Kohli, Arunima; Kawli, Trupti; Martin, Dorrelyn Patacsil; Snyder, Michael; Bacchetta, Rosa; Nadeau, Kari C.

2014-01-01

330

Identification of STAT5A and STAT5B target genes in human T cells.  

PubMed

Signal transducer and activator of transcription (STAT) comprises a family of universal transcription factors that help cells sense and respond to environmental signals. STAT5 refers to two highly related proteins, STAT5A and STAT5B, with critical function: their complete deficiency is lethal in mice; in humans, STAT5B deficiency alone leads to endocrine and immunological problems, while STAT5A deficiency has not been reported. STAT5A and STAT5B show peptide sequence similarities greater than 90%, but subtle structural differences suggest possible non-redundant roles in gene regulation. However, these roles remain unclear in humans. We applied chromatin immunoprecipitation followed by DNA sequencing using human CD4(+) T cells to detect candidate genes regulated by STAT5A and/or STAT5B, and quantitative-PCR in STAT5A or STAT5B knock-down (KD) human CD4(+) T cells to validate the findings. Our data show STAT5A and STAT5B play redundant roles in cell proliferation and apoptosis via SGK1 interaction. Interestingly, we found a novel, unique role for STAT5A in binding to genes involved in neural development and function (NDRG1, DNAJC6, and SSH2), while STAT5B appears to play a distinct role in T cell development and function via DOCK8, SNX9, FOXP3 and IL2RA binding. Our results also suggest that one or more co-activators for STAT5A and/or STAT5B may play important roles in establishing different binding abilities and gene regulation behaviors. The new identification of these genes regulated by STAT5A and/or STAT5B has major implications for understanding the pathophysiology of cancer progression, neural disorders, and immune abnormalities. PMID:24497979

Kanai, Takahiro; Seki, Scott; Jenks, Jennifer A; Kohli, Arunima; Kawli, Trupti; Martin, Dorrelyn Patacsil; Snyder, Michael; Bacchetta, Rosa; Nadeau, Kari C

2014-01-01

331

Liposomal amphotericin B as a treatment for human leishmaniasis  

PubMed Central

Introduction: Leishmaniasis is a parasitic disease transmitted by phlebotomine sandflies. Between 700,000 and 1.2 million cases of cutaneous leishmaniasis and between 200,000 and 400,000 cases of visceral leishmaniasis (VL), which is fatal if left untreated, occur annually worldwide. Liposomal amphotericin B (LAMB), alone or in combination with other drugs, has been extensively studied as VL treatment, but data on routine field use are limited, and several challenges to patients' access to this life-saving drug remain. Areas covered: This article provides a review of clinical studies on LAMB for VL and other forms of leishmaniasis. The current development of generic versions of LAMB and related challenges are also discussed. Expert opinion: LAMB proved to be highly efficacious and safe in over 8000 VL patients treated by Mdecins Sans Frontires in South Asia, and its use was feasible even at primary healthcare level. Despite requiring higher doses, LAMB is the drug of choice to treat vulnerable groups (e.g., pregnant or HIV positive) and relapsing VL patients in East Africa. LAMB should be included in national VL guidelines and registered in all VL endemic countries. Its cost should be further reduced and regulatory pathways to prove bioequivalence for generic LAMB products should be implemented. PMID:23167833

Balasegaram, Manica; Ritmeijer, Koert; Lima, Maria Angeles; Burza, Sakib; Ortiz Genovese, Gemma; Milani, Barbara; Gaspani, Sara; Potet, Julien; Chappuis, Francois

2012-01-01

332

Molecular characterization of human respiratory syncytial virus subtype B: A novel genotype of subtype B circulating in China.  

PubMed

Human respiratory syncytial virus (HRSV) is major pathogen of lower respiratory tract infections in infants and young children worldwide. There have been many studies regarding HRSV subgroup A (HRSV-A) G protein genetic variability but little information about HRSV subtype B (HRSV-B) G protein genetic diversity and molecular evolution in China. Thus, a survey of the molecular epidemiology and evolution of the G protein in China is of high importance. In this study, the circulation and genetic diversity of HRSV in Chongqing, Southwestern China, from June 2009 to May 2013, were investigated. A total of 3,167 nasopharyngeal aspirates were obtained in this study, and it was found that HRSV-B predominated in the 2009-2010 and 2012-2013 epidemic seasons. This study identified the genetic variability of the glycoprotein G gene among 102 HRSV-B strains isolated by cell culture from Chongqing nasopharyngeal aspirates, and 68 Chinese HRSV-B sequences were deposited in GenBank. Genotyping and phylogenetic analysis revealed that the HRSV-B strains were clustered into three genotypes: BA (n?=?111, 65.29%), GB3 (n?=?5, 2.94%), and a new GB genotype (n?=?54, 31.77%) named GB5. The GB5 strains varied from other genotypes in the central conserved region and N-glycosylation sites. The estimated evolutionary rate of Chinese HRSV-B was 2.01??10(-3) nucleotide substitutions/site/year, which is similar to the reports from Belgium and the Netherlands with 1.95??10(-3) and 2.78??10(-3) nucleotide substitutions/site/year, respectively. This study provides data on the circulating pattern and molecular characterization of HRSV-B genotypes in China during four consecutive years and may contribute to HRSV vaccine development. J. Med. Virol. 87: 1-9, 2015. 2014 Wiley Periodicals, Inc. PMID:24910250

Ren, Luo; Xiao, Qiuyan; Zhou, Lili; Xia, Qiuling; Liu, Enmei

2015-01-01

333

Expression of novel interleukin 2 binding molecules and their functional roles in human B cell differentiation.  

PubMed Central

Expressions and functional roles of novel IL-2 binding molecules (p70, 75) in the differentiation of B cells into Ig secreting cells were explored by using human several B cell lines and tonsillar B cells. Affinity-crosslinking studies revealed that five of nine B cell lines expressed p70 and p75 without detectable Tac antigen (p55) expression and the expression was associated with B cell maturation. In tonsillar B cells, small high-density B cells did not express p70 and p75, whereas large low-density B cells, which were thought to be activated in vivo, expressed them. Binding assays of radiolabeled IL-2 showed that the affinity of these molecules was intermediate (kD = 1-3 nM, 700-3,000 sites/cell). Furthermore, high concentrations of IL-2 (greater than 100 U/ml) induced Ig productions in large B cells and two of five cell lines. These results taken together suggest that B cells may express novel IL-2 binding molecules, associated with B cell differentiation and differentiate into Ig secreting cells by IL-2 through novel IL-2 binding molecules. Images PMID:2839549

Tanaka, T; Saiki, O; Doi, S; Suemura, M; Negoro, S; Kishimoto, S

1988-01-01

334

Stimulatory and inhibitory influences of human immunodeficiency virus on normal B lymphocytes.  

PubMed Central

B-lymphocyte dysfunction is a characteristic feature of the acquired immunodeficiency syndrome (AIDS) and of the AIDS-related complex. The aim of the present study was to further examine the influences exercised by the human immunodeficiency virus (HIV; formerly called human T-lymphotropic virus type III or lymphadenopathy-associated virus, HTLV-III/LAV) on normal human B lymphocytes. An unfractionated protein preparation, made from HIV purified by density gradient centrifugation, was previously shown to induce differentiation of normal human B lymphocytes into immunoglobulin-secreting cells. In the present analyses, this B-lymphocyte response peaked on day 6 or 7 after culture initiation and was found to be independent of the requirement for monocytes but to require T cells. Responses could also be elicited in cultures of purified B cells by the addition of T cells that had been exposed to HIV antigen. Inhibitors of protein synthesis (puromycin and cycloheximide) abrogated the responses. In contrast to its stimulatory effects, the same virus preparation was previously shown to inhibit polyclonal responses that are normally elicited in peripheral blood lymphocyte cultures by a T-dependent stimulus (pokeweed mitogen) and T-independent stimulus (Epstein-Barr virus). The present studies suggest that the inhibitory effects of the HIV antigen studied herein are targeted primarily at the B lymphocytes. The role of T lymphocytes in the HIV antigen-mediated inhibitory effects, although demonstrated, could not be conclusively established as an essential pathway. These findings elucidate mechanisms by which components of HIV exert stimulatory as well as inhibitory effects on human B lymphocytes and thereby lead to the dysfunction of these cells in HIV infection. PMID:3024167

Pahwa, S; Pahwa, R; Good, R A; Gallo, R C; Saxinger, C

1986-01-01

335

Saposin B Is a Human Coenzyme Q10-Binding/Transfer Protein  

PubMed Central

Coenzyme Q10 (CoQ10) is essential for ATP production in the mitochondria, and is an important antioxidant in every biomembrane and lipoprotein. Due to its hydrophobicity, a binding and transfer protein for CoQ10 is plausible, but none have yet been isolated and characterized. Here we purified a CoQ10-binding protein from human urine and identified it to be saposin B, a housekeeping protein necessary for sphingolipid hydrolysis in lysosomes. We confirmed that cellular saposin B binds CoQ10 in human sperm and the hepatoma cell line HepG2 by using saposin B monoclonal antibody. The molar ratios of CoQ10 to saposin B were estimated to be 0.22 in urine, 0.003 in HepG2, and 0.12 in sperm. We then confirmed that aqueous saposin B extracts CoQ10 from hexane to form a saposin B-CoQ10 complex. Lipid binding affinity to saposin B decreased in the following order: CoQ10>CoQ9>CoQ7>>?-tocopherol>>cholesterol (no binding). The CoQ10-binding affinity to saposin B increased with pH, with maximal binding seen at pH7.4. On the other hand, the CoQ10-donating activity of the saposin B-CoQ10 complex to erythrocyte ghost membranes increased with decreasing pH. These results suggest that saposin B binds and transports CoQ10 in human cells. PMID:18385835

Jin, GuangZhi; Kubo, Hiroshi; Kashiba, Misato; Horinouchi, Ryo; Hasegawa, Makoto; Suzuki, Masaru; Sagawa, Tomofumi; Oizumi, Mikiko; Fujisawa, Akio; Tsukamoto, Hideo; Yoshimura, Shinichi; Yamamoto, Yorihiro

2008-01-01

336

Failure to detect functional neutrophil B helper cells in the human spleen.  

PubMed

A novel role for human neutrophilic granulocytes was recently described, showing that these cells, upon entering the spleen, can be reprogrammed into a distinct B cell-helper neutrophil phenotype that is capable of eliciting B cell responses such as immunoglobulin secretion, class switch recombination and somatic hypermutation. Using similar protocols, we detected a homogeneous population of CD15(high)CD16(high) neutrophils in fresh human spleen samples, which did not differ in phenotype and function from blood neutrophils. No phenotypic characteristics of costimulatory nature were detected on splenic or circulating neutrophils, nor could we reproduce the immunoglobulin production of splenic B cells in the presence of splenic neutrophils, although B cell function and neutrophil activity were normal. Independent confirmation of a role for NBH cells is required. PMID:24523887

Nagelkerke, Sietse Quirijn; aan de Kerk, Daan Jacob; Jansen, Machiel Hugo; van den Berg, Timo Kars; Kuijpers, Taco Willem

2014-01-01

337

Generation of Knot Net for Calculation of Quadratic Triangular B-spline Surface of Human Head  

NASA Astrophysics Data System (ADS)

This paper deals with calculation of the quadratic triangular B-spline surface of the human head for the purpose of its modeling in the standard videocodec MPEG-4 SNHC. In connection with this we propose an algorithm of generation of the knot net and present the results of its application for triangulation of the 3D polygonal model Candide. Then for the model and generated knot net as well as an established distribution of control points we show the results of the calculated quadratic triangular B-spline surface of the human head including its textured version for the texture of the selected avatar.

Mihalk, Jn

2011-09-01

338

Oltipraz-mediated Changes in Aflatoxin B, Biotransformation in Rat Liver: Implications for Human Chemointervention1  

Microsoft Academic Search

Oltipraz (OPZ) is currently being considered for human use to protect against aflatoxin B, (AFB)-induced hepatocarcinogenesis based on its proven protective effect in rats. The effectiveness of this treatment pre sumes that orthologous cytochrome P450 and glutathione S-transferase (GST) isozymes metabolize AFB in humans as they do in rats. In this study, alterations in the expression of multiple forms of

Timo M. Buetler; Theo K. Bammler; John D. Hayes; David L. Eaton

339

Human mitochondrial transcription factor B1 methylates ribosomal RNA at a conserved stem-loop.  

PubMed

Human mitochondrial transcription factor B1 (h-mtTFB1) has an unprecedented relationship to RNA methyltransferases. Here, we show that this protein methylates a conserved stem-loop in bacterial 16S rRNA and that the homologous sequence in the human mitochondrial 12S molecule is similarly modified. Thus, h-mtTFB1 appears to be dual-function protein, acting both as a transcription factor and an rRNA-modification enzyme. PMID:12496758

Seidel-Rogol, Bonnie L; McCulloch, Vicki; Shadel, Gerald S

2003-01-01

340

Interaction of hepatitis B virus core protein with human GIPC1  

Microsoft Academic Search

Up to now, little is known about hepatitis B virus core protein (HBc) interactions with host-cell proteins, although such\\u000a interactions might be essential for virus propagation and pathogenicity. In this work, a human liver cDNA library was screened\\u000a for proteins interacting with HBc. Among several HBc-interacting partners selected, it interacted most strongly with the human\\u000a protein GIPC1. A common protein

Raimundas Razanskas; Kestutis Sasnauskas

2010-01-01

341

Inactivity of Recombinant ELA2B Provides a New Example of Evolutionary Elastase Silencing in Humans  

Microsoft Academic Search

Background: The archetypal mammalian elastase (ELA1) is not expressed in the human pancreas, because evolutionary mutations suppressed transcription of the ELA1 gene. Aims: In this study, we tested the theory that the unique duplication of the ELA2 gene in humans might compensate for the loss of ELA1. Methods: Recombinant ELA2A and ELA2B were expressed in Escherichia coli, and their activity

Mikls Sahin-Tth

2006-01-01

342

Postoligomerization folding of human cytomegalovirus glycoprotein B: identification of folding intermediates and importance of disulfide bonding.  

PubMed

Human cytomegalovirus glycoprotein B (gB or UL55) has been demonstrated to be a disulfide-linked homodimer within the envelope of mature virions. Previously, it has been shown that gB undergoes a rapid dimerization nearly coincident with its synthesis. Following dimerization, the molecule slowly folds into a form which can be transported from the endoplasmic reticulum. In this study we have examined the prolonged folding of gB by using a set of defined gB-reactive murine monoclonal antibodies and gB expressed as a recombinant protein in the absence of other human cytomegalovirus proteins. Our results have documented a folding pathway consistent with the relatively rapid dimerization of the translation product followed by delayed conversion into a fully folded molecule. Assembly of the dominant antigenic domain of gB, AD-1, preceded dimerization and folding of the molecule. The fully folded dimer was heat stable, but its conformation was altered by treatment with 2% sodium dodecyl sulfate (SDS), whereas an oligomeric folding intermediate was both heat and SDS stable. Postoligomerization disulfide bond formation could be demonstrated during folding of gB, suggesting that the formation of these covalent bonds could contribute to the prolonged folding of this glycoprotein. PMID:7474121

Billstrom, M A; Britt, W J

1995-11-01

343

Postoligomerization folding of human cytomegalovirus glycoprotein B: identification of folding intermediates and importance of disulfide bonding.  

PubMed Central

Human cytomegalovirus glycoprotein B (gB or UL55) has been demonstrated to be a disulfide-linked homodimer within the envelope of mature virions. Previously, it has been shown that gB undergoes a rapid dimerization nearly coincident with its synthesis. Following dimerization, the molecule slowly folds into a form which can be transported from the endoplasmic reticulum. In this study we have examined the prolonged folding of gB by using a set of defined gB-reactive murine monoclonal antibodies and gB expressed as a recombinant protein in the absence of other human cytomegalovirus proteins. Our results have documented a folding pathway consistent with the relatively rapid dimerization of the translation product followed by delayed conversion into a fully folded molecule. Assembly of the dominant antigenic domain of gB, AD-1, preceded dimerization and folding of the molecule. The fully folded dimer was heat stable, but its conformation was altered by treatment with 2% sodium dodecyl sulfate (SDS), whereas an oligomeric folding intermediate was both heat and SDS stable. Postoligomerization disulfide bond formation could be demonstrated during folding of gB, suggesting that the formation of these covalent bonds could contribute to the prolonged folding of this glycoprotein. PMID:7474121

Billstrom, M A; Britt, W J

1995-01-01

344

Distribution of NF-B-binding sites across human chromosome 22  

Microsoft Academic Search

We have mapped the chromosomal binding site distribution of a transcription factor in human cells. The NF-B family of transcription factors plays an essential role in regulating the induction of genes involved in several physiological processes, including apoptosis, immunity, and inflammation. The binding sites of the NF-B family member p65 were determined by using chromatin immunoprecipitation and a genomic microarray

Rebecca Martone; Ghia Euskirchen; Paul Bertone; Stephen Hartman; Thomas E. Royce; Nicholas M. Luscombe; John L. Rinn; F. Kenneth Nelson; Perry Miller; Mark Gerstein; Sherman Weissman; Michael Snyder

2003-01-01

345

Do Humans Really Learn A[superscript n] B[superscript n] Artificial Grammars from Exemplars?  

ERIC Educational Resources Information Center

An important topic in the evolution of language is the kinds of grammars that can be computed by humans and other animals. Fitch and Hauser (F&H; 2004) approached this question by assessing the ability of different species to learn 2 grammars, (AB)[superscript n] and A[superscript n] B[superscript n]. A[superscript n] B[superscript n] was taken to

Hochmann, Jean-Remy; Azadpour, Mahan; Mehler, Jacques

2008-01-01

346

A New Perspective in the Assessment of Vitamin B6 Nutritional Status during Pregnancy in Humans  

Microsoft Academic Search

Plasma pyridoxal-5'-phosphate (PLP) concentration has been suggested as a valid indicator to assess vitamin B-6 nutritionalstatus. Animal and human studies have shown that plasma PLP concentrations decrease progressively during pregnancy and large doses of vitamin B-6 supplementation are requiredto maintain plasma PLP at early or prepregnantlevels. PLPis known to be hydrolyzedto pyridoxal(PL) by alkalinephosphatase (ALP), resulting in an inverse relationshipbetween

HENDRIK C. BARNARD; WILLIAM J. H. VERMAAK; GIDEON M. POTGIETER

347

A- and B-type lamins are differentially expressed in normal human tissues  

Microsoft Academic Search

A selection of normal human tissues was investigated for the presence of lamins B1, B2, and A-type lamins, using a panel\\u000a of antibodies specific for the individual lamin subtypes. By use of immunoprecipitation and two-dimensional immunoblotting\\u000a techniques we demonstrated that these antibodies do not cross-react with other lamin subtypes and that a range of different\\u000a phosphorylation isoforms is recognized by

Jos L. V. Broers; Barbie M. Machiels; Helma J. H. Kuijpers; Frank Smedts; Ronald van den Kieboom; Yves Raymond; Frans C. S. Ramaekers

1997-01-01

348

Ex Vivo Analysis of Human Memory B Lymphocytes Specific for A and B Influenza Hemagglutinin by Polychromatic Flow-Cytometry  

PubMed Central

Understanding the impact that human memory B-cells (MBC), primed by previous infections or vaccination, exert on neutralizing antibody responses against drifted influenza hemagglutinin (HA) is key to design best protective vaccines. A major obstacle to these studies is the lack of practical tools to analyze HA-specific MBCs in human PBMCs ex vivo. We report here an efficient method to identify MBCs carrying HA-specific BCR in frozen PBMC samples. By using fluorochrome-tagged recombinant HA baits, and vaccine antigens from mismatched influenza strains to block BCR-independent binding, we developed a protocol suitable for quantitative, functional and molecular analysis of single MBCs specific for HA from up to two different influenza strains in the same tube. This approach will permit to identify the naive and MBC precursors of plasmablasts and novel MBCs appearing in the blood following infection or vaccination, thus clarifying the actual contribution of pre-existing MBCs in antibody responses against novel influenza viruses. Finally, this protocol can allow applying high throughput deep sequencing to analyze changes in the repertoire of HA+ B-cells in longitudinal samples from large cohorts of vaccinees and infected subjects with the ultimate goal of understanding the in vivo B-cell dynamics driving the evolution of broadly cross-protective antibody responses. PMID:23976947

Bardelli, Monia; Buricchi, Francesca; Tavarini, Simona; Sammicheli, Chiara; Nuti, Sandra; Degl'Innocenti, Elena; Isnardi, Isabelle; Fragapane, Elena; Del Giudice, Giuseppe; Castellino, Flora; Galli, Grazia

2013-01-01

349

Evidence for the existence of self-reactive human B lymphocytes.  

PubMed Central

Supernatants from human tonsil cells cultured in the presence of the PBAs, LPS and F (ab)2 monomers of rabbit anti-human beta2-microglobulin were found to have a complement-dependent cytotoxic activity to tonsil cells of the same donors. A direct toxic effect of the PBAs on the target cells was excluded by controls. Therefore the most likely explanation for this finding is that human lymphocytes contain a subpopulation of self-reactive B cells which can be triggered by PBAs to release antibodies to self-antigens. The implications of this finding for the understanding of infections accompanied by auto-immune phenomena are discussed. PMID:302772

Primi, D; Smith, C I; Hammarstrom, L; Lundquist, P G; Moller, G

1977-01-01

350

Oxidation of PAH trans-Dihydrodiols by Human Aldo-Keto Reductase AKR1B10  

PubMed Central

AKR1B10 has been identified as a potential biomarker for human non-small cell lung carcinoma and as a tobacco exposure and response gene. AKR1B10 functions as an efficient retinal reductase in vitro, and may regulate retinoic acid homeostasis. However, the possibility that this enzyme is able to activate polycyclic aromatic hydrocarbon (PAH) trans-dihydrodiols to form reactive and redox-active o-quinones has not been investigated to date. AKR1B10 was found to oxidize a wide range of PAH trans-dihydrodiol substrates in vitro to yield PAH o-quinones. Reactions of AKR1B10 proceeded with improper stereochemistry, since it was specific for the minor (+)-benzo[a]pyrene-7S,8S-dihydrodiol diastereomer formed in vivo. However, AKR1B10 displayed reasonable activity in the oxidation of both the (?)-R,R and (+)-S,S stereoisomers of benzo[g]chrysene-11,12-dihydrodiol and oxidized the potentially relevant, albeit minor, (+)-benz[a]anthracene-3S,4S-dihydrodiol metabolite. We find that AKR1B10 is therefore likely to play a contributing role in the activation of PAH trans-dihydrodiols in human lung. AKR1B10 retinal reductase activity was confirmed in vitro and found to be 5- to 150-fold greater than the oxidation of PAH trans-dihydrodiols examined. AKR1B10 was highly expressed at the mRNA and protein levels in human lung adenocarcinoma A549 cells, and robust retinal reductase activity was measured in lysates of these cells. The much greater catalytic efficiency of retinal reduction compared to PAH trans-dihydrodiol metabolism suggests AKR1B10 may play a greater role in lung carcinogenesis through dysregulation of retinoic acid homeostasis than through oxidation of PAH trans-dihydrodiols. PMID:18788756

Quinn, Amy M.; Harvey, Ronald G.; Penning, Trevor M.

2009-01-01

351

Association of IP-10 gene polymorphism with susceptibility to Enterovirus 71 infection  

PubMed Central

Enterovirus 71 (EV71) often causes large outbreaks of diseases among children worldwide and its pathogenesis remains unclear. The aim of the present study was to investigate the association between interferon-inducible protein 10 (IP-10) polymorphism in children with EV71 infection. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) were performed to analyze the gene polymorphisms of IP-10 (?1596C/T) in 58 EV71-infected and 48 control patients. The results showed that in EV71-infected patients the frequency of carrying CT + TT genotype and T allele is 10.3 and 6.0%, respectively, which is significantly lower than that of the controls (29.2 and 15.6%, respectively). Individuals with T allele had a lower risk of EV71 infection [odds ratio (OR) = 0.35, 95% confidence interval (CI), 0.130.89]. The results of this study indicated that ?1596T allele for the IP-10 gene may be a beneficial factor for EV71 infection. PMID:24648959

YANG, JING; CHEN, ZHEN-ZHEN; LV, TIE-GANG; LIU, PEI-PEI; CHEN, ZONG-BO

2013-01-01

352

Enterovirus 71 Induces Mitochondrial Reactive Oxygen Species Generation That is Required for Efficient Replication  

PubMed Central

Redox homeostasis is an important host factor determining the outcome of infectious disease. Enterovirus 71 (EV71) infection has become an important endemic disease in Southeast Asia and China. We have previously shown that oxidative stress promotes viral replication, and progeny virus induces oxidative stress in host cells. The detailed mechanism for reactive oxygen species (ROS) generation in infected cells remains elusive. In the current study, we demonstrate that mitochondria were a major ROS source in EV71-infected cells. Mitochondria in productively infected cells underwent morphologic changes and exhibited functional anomalies, such as a decrease in mitochondrial electrochemical potential ??m and an increase in oligomycin-insensitive oxygen consumption. Respiratory control ratio of mitochondria from infected cells was significantly lower than that of normal cells. The total adenine nucleotide pool and ATP content of EV71-infected cells significantly diminished. However, there appeared to be a compensatory increase in mitochondrial mass. Treatment with mito-TEMPO reduced eIF2? phosphorylation and viral replication, suggesting that mitochondrial ROS act to promote viral replication. It is plausible that EV71 infection induces mitochondrial ROS generation, which is essential to viral replication, at the sacrifice of efficient energy production, and that infected cells up-regulate biogenesis of mitochondria to compensate for their functional defect. PMID:25401329

Cheng, Mei-Ling; Weng, Shiue-Fen; Kuo, Chih-Hao; Ho, Hung-Yao

2014-01-01

353

Incorporating susceptible subpopulations in microbial risk assessment: pediatric exposures to enteroviruses in river water.  

PubMed

The City of Stockton, California operates a wastewater treatment facility that discharges treated effluent to the San Joaquin River. During a recent discharge permit renewal, the question was raised whether pathogenic microorganisms in the effluent may cause an unacceptably high health risk for body contact recreation in the vicinity of the discharge. An investigation was initiated to characterize the risk to public health via body contact recreation in the San Joaquin River under various flow and treatment scenarios. In this investigation, a disease transmission model was applied to quantitatively characterize the relative risk associated with various treatment and flow scenarios for the City of Stockton's wastewater treatment facility. An important component of the investigation was to assess the feasibility of quantitatively characterizing the risk to highly susceptible subpopulations for effluent-related exposures to enteroviruses. This paper presents the methods used to conduct the feasibility assessment, the conclusions drawn for this project, and our recommendations to improve exposure assessments of susceptible subpopulations' contact with microbial pathogens in recreational water. PMID:12679796

Parkin, Rebecca T; Soller, Jeffrey A; Olivieri, Adam W

2003-03-01

354

Distribution of NF-?B-binding sites across human chromosome 22  

PubMed Central

We have mapped the chromosomal binding site distribution of a transcription factor in human cells. The NF-?B family of transcription factors plays an essential role in regulating the induction of genes involved in several physiological processes, including apoptosis, immunity, and inflammation. The binding sites of the NF-?B family member p65 were determined by using chromatin immunoprecipitation and a genomic microarray of human chromosome 22 DNA. Sites of binding were observed along the entire chromosome in both coding and noncoding regions, with an enrichment at the 5? end of genes. Strikingly, a significant proportion of binding was seen in intronic regions, demonstrating that transcription factor binding is not restricted to promoter regions. NF-?B binding was also found at genes whose expression was regulated by tumor necrosis factor ?, a known inducer of NF-?B-dependent gene expression, as well as adjacent to genes whose expression is not affected by tumor necrosis factor ?. Many of these latter genes are either known to be activated by NF-?B under other conditions or are consistent with NF-?B's role in the immune and apoptotic responses. Our results suggest that binding is not restricted to promoter regions and that NF-?B binding occurs at a significant number of genes whose expression is not altered, thereby suggesting that binding alone is not sufficient for gene activation. PMID:14527995

Martone, Rebecca; Euskirchen, Ghia; Bertone, Paul; Hartman, Stephen; Royce, Thomas E.; Luscombe, Nicholas M.; Rinn, John L.; Nelson, F. Kenneth; Miller, Perry; Gerstein, Mark; Weissman, Sherman; Snyder, Michael

2003-01-01

355

Modification of substrate-inhibitor affinities of human platelet monoamine oxidase B in vitro.  

PubMed

The rate of benzylamine utilization by monoamine oxidase (MAO)-B from human blood platelets was 2-4 times higher than that for octopamine. Both activities were inhibited 100% by 10(-7) M deprenyl (a specific MAO-B inhibitor) and were not affected by clorgyline (a specific MAO-A inhibitor) or by polyclonal antibodies to MAO-A. The preincubation of platelet MAO-B with purified MAO-A from mitochondrial membranes of human placenta resulted in appearance of excess octopamine activity. This additional activity was not precipitated by antibodies to MAO-A or inhibited by deprenyl but was inhibited by clorgyline. Incubation of the MAO-A preparation from placenta at 45 degrees C for 15 min before its preincubation with MAO-B caused 50% loss of both activities. Protease inhibitors had no effect on the modification of MAO. These data indicate that MAO-A or a factor tightly bound to it can modify MAO-B yielding a form of the enzyme with both MAO-A and MAO-B substrate and inhibitor affinities and MAO-B immunospecificity. PMID:2509446

Szutowicz, A; Tomaszewicz, M; Orsulak, P J

1989-10-25

356

Transgenic rice containing human CYP2B6 detoxifies various classes of herbicides.  

PubMed

The human gene for CYP2B6, a cytochrome P450 monooxygenase that inactivates xenobiotic chemicals, was introduced into Oryza sativa cv. Nipponbare by Agrobacterium-mediated transformation. At germination, R(1) seeds of transgenic rice plants expressing CYP2B6 (CYP2B6 rice) showed a high tolerance to 5 microM metolachlor, a preemergence herbicide that is degraded by CYP2B6. Thin-layer chromatography after culture with (14)C-labeled metolachlor revealed that the amounts of residual metolachlor decreased in plant tissues and the medium of CYP2B6 rice faster than those of untransformed Nipponbare. CYP2B6 rice plants were able to grow in the presence of 13 out of 17 herbicides: five chloroacetamides and mefenacet, pyributicarb, amiprofos-methyl, trifluralin, pendimethalin, norflurazon, and chlorotoluron. These herbicides differ in their modes of action and chemical structures. Transgenic rice expressing a xenobiotic-degrading human CYP2B6, which has broad substrate specificity, should be good not only for developing herbicide tolerant rice but also for reducing the environmental impact of agrochemicals. PMID:15853388

Hirose, Sakiko; Kawahigashi, Hiroyuki; Ozawa, Kenjirou; Shiota, Noriaki; Inui, Hideyuki; Ohkawa, Hideo; Ohkawa, Yasunobu

2005-05-01

357

Plasmacytoid dendritic cells promote rotavirus-induced human and murine B cell responses  

PubMed Central

B celldependent immunity to rotavirus, an important intestinal pathogen, plays a significant role in viral clearance and protects against reinfection. Human in vitro and murine in vivo models of rotavirus infection were used to delineate the role of primary plasmacytoid DCs (pDCs) in initiating B cell responses. Human pDCs were necessary and sufficient for B cell activation induced by rotavirus. Type I IFN recognition by B cells was essential for rotavirus-mediated B cell activation in vitro and murine pDCs and IFN-?/?mediated B cell activation after in vivo intestinal rotavirus infection. Furthermore, rotavirus-specific serum and mucosal antibody responses were defective in mice lacking functional pDCs at the time of infection. These data demonstrate that optimal B cell activation and virus-specific antibody secretion following mucosal infection were a direct result of pDC-derived type I IFN. Importantly, viral shedding significantly increased in pDC-deficient mice, suggesting that pDC-dependent antibody production influences viral clearance. Thus, mucosal pDCs critically influence the course of rotavirus infection through rotavirus recognition and subsequent IFN production and display powerful adjuvant properties to initiate and enhance humoral immunity. PMID:23635775

Deal, Emily M.; Lahl, Katharina; Narvaez, Carlos F.; Butcher, Eugene C.; Greenberg, Harry B.

2013-01-01

358

BDNF/TrkB signaling protects HT-29 human colon cancer cells from EGFR inhibition  

SciTech Connect

Highlights: Black-Right-Pointing-Pointer BDNF protected HT-29 colorectal cancer cells from the antitumor effect of cetuximab. Black-Right-Pointing-Pointer TrkB inhibition potentiated the antitumor effect of cetuximab. Black-Right-Pointing-Pointer BDNF/TrkB signaling might be involved in resistance to anti-EGFR therapy. -- Abstract: The clinical success of targeted treatment of colorectal cancer (CRC) is often limited by resistance to anti-epidermal growth factor receptor (EGFR) therapy. The neurotrophin brain-derived neurotrophic factor (BDNF) and its receptor TrkB have recently emerged as anticancer targets, and we have previously shown increased BDNF levels in CRC tumor samples. Here we report the findings from in vitro experiments suggesting that BDNF/TrkB signaling can protect CRC cells from the antitumor effects of EGFR blockade. The anti-EGFR monoclonal antibody cetuximab reduced both cell proliferation and the mRNA expression of BDNF and TrkB in human HT-29 CRC cells. The inhibitory effect of cetuximab on cell proliferation and survival was counteracted by the addition of human recombinant BDNF. Finally, the Trk inhibitor K252a synergistically enhanced the effect of cetuximab on cell proliferation, and this effect was blocked by BDNF. These results provide the first evidence that increased BDNF/TrkB signaling might play a role in resistance to EGFR blockade. Moreover, it is possible that targeting TrkB could potentiate the anticancer effects of anti-EGFR therapy.

Brunetto de Farias, Caroline [Cancer Research Laboratory, University Hospital Research Center (CPE-HCPA), Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS (Brazil) [Cancer Research Laboratory, University Hospital Research Center (CPE-HCPA), Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS (Brazil); Children's Cancer Institute, 90420-140 Porto Alegre, RS (Brazil); Laboratory of Neuropharmacology and Neural Tumor Biology, Department of Pharmacology, Institute for Basic Health Sciences, Federal University of Rio Grande do Sul, 90050-170 Porto Alegre, RS (Brazil); National Institute for Translational Medicine (INCT-TM), 90035-003 Porto Alegre, RS (Brazil); Heinen, Tiago Elias; Pereira dos Santos, Rafael [Cancer Research Laboratory, University Hospital Research Center (CPE-HCPA), Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS (Brazil) [Cancer Research Laboratory, University Hospital Research Center (CPE-HCPA), Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS (Brazil); Laboratory of Neuropharmacology and Neural Tumor Biology, Department of Pharmacology, Institute for Basic Health Sciences, Federal University of Rio Grande do Sul, 90050-170 Porto Alegre, RS (Brazil); National Institute for Translational Medicine (INCT-TM), 90035-003 Porto Alegre, RS (Brazil); Abujamra, Ana Lucia [Cancer Research Laboratory, University Hospital Research Center (CPE-HCPA), Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS (Brazil) [Cancer Research Laboratory, University Hospital Research Center (CPE-HCPA), Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS (Brazil); Children's Cancer Institute, 90420-140 Porto Alegre, RS (Brazil); National Institute for Translational Medicine (INCT-TM), 90035-003 Porto Alegre, RS (Brazil); Schwartsmann, Gilberto [Cancer Research Laboratory, University Hospital Research Center (CPE-HCPA), Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS (Brazil) [Cancer Research Laboratory, University Hospital Research Center (CPE-HCPA), Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS (Brazil); National Institute for Translational Medicine (INCT-TM), 90035-003 Porto Alegre, RS (Brazil); Department of Internal Medicine, School of Medicine, Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS (Brazil); and others

2012-08-24

359

Of mice and men: Different functions of the murine and human 2B4 (CD244) receptor on NK cells  

Microsoft Academic Search

2B4 was initially discovered on murine NK cells and T cells displaying non-MHC dependent cytotoxicity. Human 2B4 was cloned based on sequence homology with mouse 2B4. Recent evidence suggests that the function of this receptor might be different in the two species. Human 2B4 activates NK cell cytotoxicity and interferon gamma production when engaged by CD48, its ligand, on target

Swapnil V. Vaidya; Porunelloor A. Mathew

2006-01-01

360

Suppression of Angiogenesis, Tumorigenicity, and Metastasis by Human Prostate Cancer Cells Engineered to Produce Interferon-b1  

Microsoft Academic Search

We determined whether the IFN-b gene can be used to suppress angiogenesis, tumor growth, and metastasis of human prostate cancer cells growing in the prostate of nude mice. Highly metastatic PC-3M human prostate cancer cells were engineered to constitutively produce murine IFN-b subsequent to infection with a retroviral vector containing murine IFN-b cDNA. Parental (PC-3M-P), control vector-transduced (PC-3M-Neo), and IFN-b-transduced

Zhongyun Dong; Graham Greene; Curtis Pettaway; Colin P. N. Dinney; Ines Eue; Weixin Lu; Corazon D. Bucana; Mevlana D. Balbay; Diane Bielenberg; Isaiah J. Fidler

361

Anti-proliferation effect of Hevea brasiliensis latex B-serum on human breast epithelial cells.  

PubMed

The rubber tree (Hevea brasiliensis) extracts are becoming increasingly visible in pharmaceutical and therapeutical research. The present study is aimed at examining the specific anti-proliferation property of H. brasiliensis latex B-serum sub-fractions against human breast cancer epithelial cell lines MCF-7 and MDA-MB231. The results showed that the latex whole B-serum and DBP sub-fraction exerted a specific anti-proliferation activity against cancer-origin cells MDA-MB231 but had little effect on non-cancer-origin cells. On the other hand, the anti-proliferative activity was diminished in the pre-heated B-serum fractions. With the low toxicity that the B-serum demonstrated previously in Brine Shrimp Lethality Test (BSLT), the present results suggest the potential use of the B-serum sub-fractions in cancer treatment. PMID:22713955

Lee, Yang Kok; Lay, Lam Kit; Mahsufi, Mansor Sharif; Guan, Teoh Siang; Elumalai, Sunderasan; Thong, Ong Ming

2012-07-01

362

Identification of a novel human leukocyte antigen allele B?46:34.  

PubMed

A new human leukocyte antigen (HLA) B allele was found in a healthy male Chinese Kazak individual. Sequencing-based typing (SBT) was used to identify and analyze the difference between the new allele and the closest matching HLA-B allele. HLA-B(?)46 new allele has 1nt change from B(?)46:01:01 at nt 853 where G->C (condon 260 GTA->CTA), resulting in a coding change: 260 Val is changed to Leu. The new HLA-B(?)46:34 allele was identified, and was named officially by the World Health Organization (WHO) Nomenclature Committee in June 2012. The GenBank sequence accession number is JX035785. PMID:23137878

Shen, Chun-mei; Wang, Hong-dan; Deng, Ya-jun; Wang, Shi-ying; Zhang, Li-ping; Zhang, Bo; Pu, Hong-wei; Wu, Qiang-ju; Liu, Wen-juan; Zhu, Bo-feng

2013-02-01

363

How robust is human gait to muscle weakness? Marjolein M. van der Krogt a,b,  

E-print Network

. Disuse, aging, and disease can lead to a loss of muscle strength, which may limit the performance movements requiring agility, timing, and strength. Disuse, aging, and disease can lead to a loss of muscleHow robust is human gait to muscle weakness?§ Marjolein M. van der Krogt a,b, *, Scott L. Delp c

Delp, Scott

364

Original Articles Identification of B and T Cells in Human Spleen  

E-print Network

Original Articles Identification of B and T Cells in Human Spleen Sections by Infrared spleen. A secondary follicle containing a germinal center and a T zone were studied in more detail, arteries, and spleen red pulp. The assignments could be confirmed in consec- utive sections

Meyer-Hermann, Michael

365

Human Resource Services -Payroll 1265 Military Trail, Room B526C/D  

E-print Network

(Dec 2013) Human Resource Services - Payroll 1265 Military Trail, Room B526C/D Toronto, Ontario, M1: Order No.: CF: Fund No.: Monthly # Of Months Total for this contract Hours Worked x = Salary $ x = $ TOTAL SALARY (EXCLUDING Vacation Pay) Chair's Name (Please Print) Telephone No. Chair's Signature Date

Boonstra, Rudy

366

Human Resource Services -Payroll 1265 Military Trail, Room B526C/D  

E-print Network

(Dec 2012) Human Resource Services - Payroll 1265 Military Trail, Room B526C/D Toronto, Ontario, M1: Order No.: CF: Fund No.: Monthly # Of Months Hours Worked Total for this contract x = Salary $ x = $ TOTAL SALARY (EXCLUDING Vacation Pay) Chair's Name (Please Print) Telephone No. Chair's Signature Date

Kronzucker, Herbert J.

367

Part B -Page 1 of 45 HUMAN RESOURCES AND MOBILITY (HRM) ACTIVITY  

E-print Network

CODY Part B - Page 1 of 45 STARTPAGE HUMAN RESOURCES AND MOBILITY (HRM) ACTIVITY MARIE CURIE .......................................... 18 2.4 Planned recruitment of Early Stage and Experienced Researchers .......... 18 2.4.1 168 months.1.6 Allocation of responsibilities ............................................. 34 4.2 Recruitment, appointment

d'Orléans, Université

368

B-SUB: A Practical Bloom-Filter-Based Publish-Subscribe System for Human Networks  

E-print Network

tolerant networks, human networks, social network analysis. I. INTRODUCTION The fast paced development querying the TCBFs, which is simple and fast. These designs make B-SUB pretty suitable for resource to the direct correspondence between such applications and people's lives, these applications are able

Wu, Jie

369

A Flexible Model For Human Circadian Rhythms Yuedong Wang and Morton B. Brown  

E-print Network

A Flexible Model For Human Circadian Rhythms Yuedong Wang and Morton B. Brown Department. In this paper we describe a semi­parametric periodic spline function that can be fit to circadian rhythms physiological processes vary in a circadian pattern. Although a sine/cosine function can be used to model

Wang, Yuedong

370

Effects of Ultraviolet B Light on Cutaneous Immune Responses of Humans with Deeply Pigmented Skin  

Microsoft Academic Search

The incidence of skin cancers of the basal and squamous cell types is extremely low among genetically black-skinned human beings, whereas these types of skin cancers are common among Caucasians, especially those who live in geographic areas of high sun exposure. Ultraviolet B light (UVB) is thought to be the primary oncogenic agent in sunlight. We have recently demonstrated that

Martin Vermeer; George J. Schmieder; Takeshi Yoshikawa; Jan-Willem van den Berg; Michael S. Metzman; J. Richard Taylor; J. Wayne Streilein

1991-01-01

371

Humanized mice with ectopic artificial liver tissues Alice A. Chena,b,c  

E-print Network

Humanized mice with ectopic artificial liver tissues Alice A. Chena,b,c , David K. Thomasd University, 29 Oxford Street, Cambridge, MA 02138; c David H. Koch Institute for Integrative Cancer Research Palliative Care, Dana-Farber Cancer Institute and Harvard Medical School, 44 Binney Street, Boston, MA 02115

Bhatia, Sangeeta

372

Nerve growth factor triggers microfilament assembly and paxillin phosphorylation in human B lymphocytes  

Microsoft Academic Search

Summary Increasing evidence suggests that the nervous system is involved in allergic inflammation. One of the potential regulatory molecules of the neuroimmune system is nerve growth factor (NGF). Recent studies from our group demonstrated the presence of a functional NGF receptor (NGFR) on human B lymphocytes. Moreover, we showed that gp140 trk tyrosine kinase, which serves as an NGFR, was

I. Melamed; C. E. Turner; K. Aktories; D. R. Kaplan

1995-01-01

373

Mutations in TNFRSF13B encoding TACI are associated with common variable immunodeficiency in humans  

Microsoft Academic Search

The functional interaction of BAFF and APRIL with TNF receptor superfamily members BAFFR, TACI and BCMA is crucial for development and maintenance of humoral immunity in mice and humans. Using a candidate gene approach, we identified homozygous and heterozygous mutations in TNFRSF13B, encoding TACI, in 13 individuals with common variable immunodeficiency. Homozygosity with respect to mutations causing the amino acid

U Salzer; H M Chapel; A D B Webster; Q Pan-Hammarstrm; A Schmitt-Graeff; M Schlesier; H H Peter; J K Rockstroh; P Schneider; A A Schffer; L Hammarstrm; B Grimbacher

2005-01-01

374

B Vitamins and n3 Fatty Acids for Brain Development and Function: Review of Human Studies  

Microsoft Academic Search

Background: Nutrition is one of many factors that affect brain development and functioning, and in recent years the role of certain nutrients has been investigated. B vitamins and n3 polyunsaturated fatty acids (PUFA) are two of the most promising and widely studied nutritional factors. Methods: In this review, we provide an overview of human studies published before August 2011 on

Ondine van de Rest; Lenneke W. A. van Hooijdonk; Esme Doets; Olga J. G. Schiepers; Ans Eilander; Lisette C. P. G. M. de Groot

2012-01-01

375

Universal dimensions of human mate preferences Todd K. Shackelford a,*, David P. Schmitt b  

E-print Network

Universal dimensions of human mate preferences Todd K. Shackelford a,*, David P. Schmitt b , David M. Buss c a Florida Atlantic University, Department of Psychology, 2912 College Avenue, Davie, FL [Buss, D. M. (1989)]. Participants from each culture responded to the same 18-item measure. Statistical

Pillow, Jonathan

376

Phospholipase B activity enhances adhesion of Cryptococcus neoformans to a human lung epithelial cell line  

Microsoft Academic Search

Secreted phospholipase B (PLB1), which contains three enzyme activities in the one protein, is necessary for the initiation of pulmonary infection by Cryptococcus neoformans and for dissemination from the lung via the lymphatics and blood. Adhesion to lung epithelium is the first step in this process, therefore we investigated the role of PLB1 in adhesion to a human lung epithelial

Ranjini Ganendren; Elizabeth Carter; Tania Sorrell; Fred Widmer; Lesley Wright

2006-01-01

377

Evaluation of organic anion transporting polypeptide 1B1 and 1B3 humanized mice as a translational model to study the pharmacokinetics of statins.  

PubMed

Organic anion transporting polypeptide (Oatp) 1a/1b knockout and OATP1B1 and -1B3 humanized mouse models are promising tools for studying the roles of these transporters in drug disposition. Detailed characterization of these models will help to better understand their utility for predicting clinical outcomes. To advance this approach, we carried out a comprehensive analysis of these mouse lines by evaluating the compensatory changes in mRNA expression, quantifying the amounts of OATP1B1 and -1B3 protein by liquid chromatography-tandem mass spectrometry, and studying the active uptake in isolated hepatocytes and the pharmacokinetics of some prototypical substrates including statins. Major outcomes from these studies were 1) mostly moderate compensatory changes in only a few genes involved in drug metabolism and disposition, 2) a robust hepatic expression of OATP1B1 and -1B3 proteins in the respective humanized mouse models, and 3) functional activities of the human transporters in hepatocytes isolated from the humanized models with several substrates tested in vitro and with pravastatin in vivo. However, the expression of OATP1B1 and -1B3 in the humanized models did not significantly alter liver or plasma concentrations of rosuvastatin and pitavastatin compared with Oatp1a/1b knockout controls under the conditions used in our studies. Hence, although the humanized OATP1B1 and -1B3 mice showed in vitro and/or in vivo functional activity with some statins, further characterization of these models is required to define their potential use and limitations in the prediction of drug disposition and drug-drug interactions in humans. PMID:24855184

Salphati, Laurent; Chu, Xiaoyan; Chen, Liangfu; Prasad, Bhagwat; Dallas, Shannon; Evers, Raymond; Mamaril-Fishman, Donna; Geier, Ethan G; Kehler, Jonathan; Kunta, Jeevan; Mezler, Mario; Laplanche, Loic; Pang, Jodie; Rode, Anja; Soars, Matthew G; Unadkat, Jashvant D; van Waterschoot, Robert A B; Yabut, Jocelyn; Schinkel, Alfred H; Scheer, Nico

2014-08-01

378

Quantitation of human parvovirus B19 DNA in erythema infectiosum and aplastic crisis.  

PubMed

Several publications concerning the methods of real-time PCR for human parvovirus B19 (B19V) have appeared and some case reports mention B19V DNA loads. However, no large-scale study quantitating levels of B19V DNA in common or representative B19V manifestations such as erythema infectiosum and aplastic crisis has been performed. Consequently, using the TaqMan PCR assay, the B19V load in a large sample of subjects with erythema infectiosum or aplastic crisis was quantitated. Sixty-five subjects in the acute phase of erythema infectiosum were involved, and in addition 22 serum samples from seven subjects with B19V-associated aplastic crisis complicating chronic hemolytic anemia were also analyzed. In the acute phase of erythema infectiosum the median B19V DNA load in the serum samples from the acute phase of erythema infectiosum was 7.63??10(5) ?genomes/ml, (range from 4.48??10(3) to 8.31??10(6) ?genomes/ml). The serum B19V DNA load during the acute phase of aplastic crisis complicating chronic hemolytic anemia was extremely high, that is 10(10) -10(13) ?genomes/ml, and decreased gradually to around 10(5) genomes/ml over 1-2 months. Although all subjects followed an almost uniform and typical clinical course of erythema infectiosum, there was a large individual variation of B19V DNA loads, that is differences of over 1,000 times. Extremely high B19V loads were observed in subjects with aplastic crisis. This study is the first large scale report of studies of the B19V DNA loads in subjects with erythema infectiosum and aplastic crisis, the most common and significant clinical manifestations by B19V infections. J. Med. Virol. 86:2102-2106, 2014. 2014 Wiley Periodicals, Inc. PMID:24962467

Ishikawa, Aki; Yoto, Yuko; Tsugawa, Takeshi; Tsutsumi, Hiroyuki

2014-12-01

379

Cathepsin B Degrades Amyloid-? in Mice Expressing Wild-type Human Amyloid Precursor Protein*  

PubMed Central

Accumulation of amyloid-? (A?), believed to be a key trigger of Alzheimer disease (AD), could result from impaired clearance mechanisms. Previously, we showed that the cysteine protease cathepsin B (CatB) degrades A?, most likely by C-terminal truncation, in mice expressing human amyloid precursor protein with familial AD-linked mutations (hAPPFAD). In addition, the A?-degrading activity of CatB is inhibited by its endogenous inhibitor, cystatin C (CysC). Reducing CysC expression markedly lowers A? levels by enhancing CatB-mediated A? degradation in hAPPFAD mice. However, because a vast majority of AD patients do not carry familial mutations, we investigated how the CysC-CatB axis affects A? levels in mice expressing wild-type hAPP (hAPPWT). Enhancing CatB activity by CysC deletion significantly lowered total A? and A?42 levels in hAPPWT mice, whereas CatB deletion increased A? levels. To determine whether neuron-derived CatB degrades A? in vivo, we generated transgenic mice overexpressing CatB under the control of a neuron-specific enolase promoter. Enhancing neuronal CatB activity in hAPPWT mice significantly lowered A?42 levels. The processing of hAPPWT was unaffected by increasing or ablating CatB activity. Thus, the CysC-CatB axis affects degradation of A?42 derived from hAPP lacking familial mutations. These findings support the notion that enhancing CatB activity could lower A?, especially A?42, in AD patients with or without familial mutations. PMID:23024364

Wang, Chao; Sun, Binggui; Zhou, Yungui; Grubb, Anders; Gan, Li

2012-01-01

380

Bordetella pertussis, the Causative Agent of Whooping Cough, Evolved from a Distinct, Human-Associated Lineage of B. bronchiseptica  

PubMed Central

Bordetella pertussis, B. bronchiseptica, B. parapertussishu, and B. parapertussisov are closely related respiratory pathogens that infect mammalian species. B. pertussis and B. parapertussishu are exclusively human pathogens and cause whooping cough, or pertussis, a disease that has resurged despite vaccination. Although it most often infects animals, infrequently B. bronchiseptica is isolated from humans, and these infections are thought to be zoonotic. B. pertussis and B. parapertussishu are assumed to have evolved from a B. bronchisepticalike ancestor independently. To determine the phylogenetic relationships among these species, housekeeping and virulence genes were sequenced, comparative genomic hybridizations were performed using DNA microarrays, and the distribution of insertion sequence elements was determined, using a collection of 132 strains. This multifaceted approach distinguished four complexes, representing B. pertussis, B. parapertussishu, and two distinct B. bronchiseptica subpopulations, designated complexes I and IV. Of the two B. bronchiseptica complexes, complex IV was more closely related to B. pertussis. Of interest, while only 32% of the complex I strains were isolated from humans, 80% of the complex IV strains were human isolates. Comparative genomic hybridization analysis identified the absence of the pertussis toxin locus and dermonecrotic toxin gene, as well as a polymorphic lipopolysaccharide biosynthesis locus, as associated with adaptation of complex IV strains to the human host. Lipopolysaccharide structural diversity among these strains was confirmed by gel electrophoresis. Thus, complex IV strains may comprise a human-associated lineage of B. bronchiseptica from which B. pertussis evolved. These findings will facilitate the study of pathogen host-adaptation. Our results shed light on the origins of the disease pertussis and suggest that the association of B. pertussis with humans may be more ancient than previously assumed. PMID:16389302

Diavatopoulos, Dimitri A; Cummings, Craig A; Schouls, Leo M; Brinig, Mary M; Relman, David A; Mooi, Frits R

2005-01-01

381

Atrial fibrillation and sinus node dysfunction in human ankyrin-B syndrome: a computational analysis  

PubMed Central

Ankyrin-B is a multifunctional adapter protein responsible for localization and stabilization of select ion channels, transporters, and signaling molecules in excitable cells including cardiomyocytes. Ankyrin-B dysfunction has been linked with highly penetrant sinoatrial node (SAN) dysfunction and increased susceptibility to atrial fibrillation. While previous studies have identified a role for abnormal ion homeostasis in ventricular arrhythmias, the molecular mechanisms responsible for atrial arrhythmias and SAN dysfunction in human patients with ankyrin-B syndrome are unclear. Here, we develop a computational model of ankyrin-B dysfunction in atrial and SAN cells and tissue to determine the mechanism for increased susceptibility to atrial fibrillation and SAN dysfunction in human patients with ankyrin-B syndrome. Our simulations predict that defective membrane targeting of the voltage-gated L-type Ca2+ channel Cav1.3 leads to action potential shortening that reduces the critical atrial tissue mass needed to sustain reentrant activation. In parallel, increased fibrosis results in conduction slowing that further increases the susceptibility to sustained reentry in the setting of ankyrin-B dysfunction. In SAN cells, loss of Cav1.3 slows spontaneous pacemaking activity, whereas defects in Na+/Ca2+ exchanger and Na+/K+ ATPase increase variability in SAN cell firing. Finally, simulations of the intact SAN reveal a shift in primary pacemaker site, SAN exit block, and even SAN failure in ankyrin-B-deficient tissue. These studies identify the mechanism for increased susceptibility to atrial fibrillation and SAN dysfunction in human disease. Importantly, ankyrin-B dysfunction involves changes at both the cell and tissue levels that favor the common manifestation of atrial arrhythmias and SAN dysfunction. PMID:23436330

Wolf, Roseanne M.; Glynn, Patric; Hashemi, Seyed; Zarei, Keyan; Mitchell, Colleen C.; Anderson, Mark E.; Mohler, Peter J.

2013-01-01

382

TLR4 activates NF-{kappa}B in human ovarian granulosa tumor cells  

SciTech Connect

Highlights: {yields} TLR4 is expressed in human ovarian granulosa tumor cells. {yields} Acting through TLR4, LPS and HSP60 induce a NF{kappa}B signaling cascade in human ovarian granulosa tumor cells. {yields} NF{kappa}B activation or inhibition did not alter chemosensitivity to TRAIL or cisplatin. -- Abstract: Previous studies have demonstrated expression of Toll-like receptors (TLRs) in the surface epithelium of normal ovaries (OSE) and in epithelial ovarian tumors. Most notably, OSE-derived cancers express TLR4, which activates the nuclear factor-kappa B (NF-{kappa}B) signaling cascade as a mediator of inflammatory response. Currently, there is considerable interest in elucidating the role of TLR-mediated signaling in cancers. Nevertheless, the expression of TLRs in granulosa cell tumors (GCTs) of the ovary, and the extent to which GCT expression of TLRs may influence cell-signaling pathways and/or modulate the efficacy of chemotherapeutics, has yet to be determined. In the present study, human GCT lines (COV434 and KGN) were utilized to evaluate expression of functional TLR4. TLR4 is expressed in GCT cell lines and ligation of TLR4 with bacterial lipopolysaccharide (LPS) led to I{kappa}B degradation and activation of NF-{kappa}B. NF-{kappa}B activation was confirmed by nuclear localization of NF-{kappa}B p65 following treatment with LPS and the naturally occurring ligand, HSP60. Notably, immunoneutralization of TLR4 blocked nuclear localization, and inhibition of NF-{kappa}B signaling attenuated LPS-induced TNF{alpha} plus increased doubling time in both cell lines. Contradictory to reports using human OSE cell lines, inhibition of NF-{kappa}B signaling failed to sensitize GCT lines to TRAIL or cisplatin. In summary, findings herein are the first to demonstrate a functional TLR-signaling pathway specifically in GCTs, and indicate that in contrast to OSE-derived cancers, inhibition of NF-{kappa}B does not sensitize GCTs to TRAIL or cisplatin.

Woods, Dori C., E-mail: dwoods2@partners.org [Vincent Center for Reproductive Biology, Vincent Obstetrics and Gynecology Service, Massachusetts General Hospital/Harvard Medical School, Boston, MA 02114 (United States); White, Yvonne A.R. [Vincent Center for Reproductive Biology, Vincent Obstetrics and Gynecology Service, Massachusetts General Hospital/Harvard Medical School, Boston, MA 02114 (United States)] [Vincent Center for Reproductive Biology, Vincent Obstetrics and Gynecology Service, Massachusetts General Hospital/Harvard Medical School, Boston, MA 02114 (United States); Dau, Caroline [University of California, San Francisco, School of Dentistry, San Francisco, CA 94143 (United States)] [University of California, San Francisco, School of Dentistry, San Francisco, CA 94143 (United States); Johnson, A.L. [Center for Reproductive Biology and Health, The Pennsylvania State University, University Park, PA 16802 (United States)] [Center for Reproductive Biology and Health, The Pennsylvania State University, University Park, PA 16802 (United States)

2011-06-17

383

The effects of vitamins C and B12 on human nasal ciliary beat frequency  

PubMed Central

Background This study was designed to investigate the effects of the vitamins C and B12 on the regulation of human nasal ciliary beat frequency (CBF). Methods Human nasal mucosa was removed endoscopically and nasal ciliated cell culture was established. Changes of CBF in response to different concentrations of vitamin C or vitamin B12 were quantified by using high-speed (240 frames per second) digital microscopy combined with a beat-by-beat CBF analysis. Results At the concentrations of 0.01% and 0.10%, vitamin C induced an initial increase, followed by a gradual decrease of CBF to the baseline level, while 1.00% vitamin C induced a reversible decrease of CBF. Vitamin B12, at the concentrations of 0.01% and 0.10%, did not influence CBF during the 20-min observation period, while a 1.00% vitamin B12 treatment caused a time-dependent but reversible decrease of CBF. Conclusions Treatment with vitamin C or vitamin B12 caused a concentration-dependent but reversible decrease of CBF in cultured human nasal epithelial cells. Therefore, it is necessary to choose a concentration that is safe, effective, and non-ciliotoxic when applying these drugs topically in the nasal cavity. PMID:23688196

2013-01-01

384

Distribution of lipid-binding regions in human apolipoprotein B-100.  

PubMed

The distribution of lipid-binding regions of human apolipoprotein B-100 has been investigated by recombining proteolytic fragments of B-100 with lipids and characterizing the lipid-bound fragments by peptide mapping, amino acid sequencing, and immunoblotting. Fragments of B-100 were generated by digestion of low-density lipoproteins (LDL) in the presence of sodium decyl sulfate with either Staphylococcus aureus V8 protease, pancreatic elastase, or chymotrypsin. Particles with electron microscopic appearance of native lipoproteins formed spontaneously when detergent was removed by dialysis from enzyme digests containing fragments of B-100 and endogenous lipids, or from incubation mixtures of delipidated B-100 fragments mixed with microemulsions of exogenous lipids (cholesteryl oleate and egg phosphatidylcholine). Fractionation of the recombinant particles by isopycnic or density gradient ultracentrifugation yielded complexes similar to native LDL with respect to shape, diameter, electrophoretic mobility, and surface and core compositions. Circular dichroic spectra of these particles showed helicity similar to LDL but a somewhat decreased content of beta-structure. Most of the fragments of B-100 were capable of binding to lipids; 12 were identified by direct sequence analysis and 14 by reaction with antisera against specific sequences within B-100. Our results indicate that lipid-binding regions of B-100 are widely distributed within the protein molecule and that proteolytic fragments derived from B-100 can reassociate in vitro with lipids to form LDL-like particles. PMID:2730877

Chen, G C; Hardman, D A; Hamilton, R L; Mendel, C M; Schilling, J W; Zhu, S; Lau, K; Wong, J S; Kane, J P

1989-03-21

385

TLR-mediated STAT3 and ERK activation controls IL-10 secretion by human B cells.  

PubMed

IL-10-producing B cells have a regulatory effect in various mouse models for immune-mediated disorders via secretion of IL-10, a potent immunoregulatory cytokine. However, currently, the signaling pathways that regulate IL-10 production in B cells are not well understood. Here, we show that TLR signaling, but not BCR activation or CD40 ligation, induces potent production of IL-10 in human B cells. We demonstrate that the activation of STAT3 and ERK is required for TLR-induced IL-10 production by B cells, since inhibition of STAT3 or ERK activation abrogates TLR-induced IL-10 production. We also uncover a novel function of the TLR-MyD88-STAT3 pathway in B cells, namely controlling IL-10 production, in addition to the known role for this pathway in antibody production. Furthermore, IFN-?, a member of the type I IFN family, differentially modulates TLR7/8- and TLR9-activated STAT3 and ERK in B cells, which provides an explanation for our findings that IFN-? enhances TLR7/8-induced, but not TLR9-induced IL-10 production. These results yield insights into the mechanisms by which TLR signaling regulates IL-10 production in B cells and how type I IFN modulates TLR-mediated IL-10 production by B cells, therefore providing potential targets to modulate the function of IL-10-producing B cells. PMID:24737107

Liu, Bi-Sheng; Cao, Yonghao; Huizinga, Tom W; Hafler, David A; Toes, Rene E M

2014-07-01

386

Hepatitis B Antigen: Antigenic Sites Related to Human Serum Proteins Revealed by Affinity Chromatography  

PubMed Central

Hepatitis B antigen-associated particles, isolated from sera of antigen carriers, were submitted to affinity chromatography on columns of insolubilized antibodies to normal human plasma. The particles adsorbed to the immunosorbent at pH 7.2 and were subsequently eluted at pH 2.2. Exposure of the particles to 8 M urea, 5 M KI, pH 2.2, detergents, organic solvents, or proteolytic enzymes failed to prevent their subsequent adsorption to the immunosorbent. This suggests that antigenic determinants related to human plasma proteins are constituent components of hepatitis B antigen-associated particles. These determinants are distinct from the group-specific (a) and subtype-specific (d or y) sites of the hepatitis B antigen and appear to be related to antigenic specificities on prealbumin, albumin, apolipoproteins C and D, and the ?-chain of immunoglobulin G. PMID:4136767

Neurath, A. Robert; Prince, Alfred M.; Lippin, Arnold

1974-01-01

387

Physical mapping of human myosin-IXB (MYO9B), the human orthologue of the rat myosin myr 5, to chromosome 19p13.1  

SciTech Connect

The gene for human myosin-IXB (MYO9B) was mapped to human chromosome 19p13.1 using fluorescence in situ hybridization. The rat myosin myr 5 gene orthologue was used for the genetic mapping of the human gene. 14 refs., 1 fig.

Baehler, M.; Kehrer, I.; Stoeffler, H.E. [Friedrich-Miescher-Laboratorium, Tuebingen (Germany)] [Friedrich-Miescher-Laboratorium, Tuebingen (Germany); Gorden, L.; Olsen, A.S. [Lawrence Livermore National Lab., CA (United States)] [Lawrence Livermore National Lab., CA (United States)

1997-07-01

388

Oligonucleotide fingerprint analysis of enterovirus 70 isolates from the 1980 to 1981 pandemic of acute hemorrhagic conjunctivitis: evidence for a close genetic relationship among Asian and American strains.  

PubMed

Enterovirus 70 isolates obtained in Asia and the Americas between 1980 and 1981 from cases of acute hemorrhagic conjunctivitis were found to be very closely related by RNase T1 oligonucleotide fingerprinting. Two closely related isolates from the first acute hemorrhagic conjunctivitis epidemic (1969 to 1972) differed by many oligonucleotides from the 1980 to 1981 pandemic strains. The strong similarities of oligonucleotide patterns of isolates from the same epidemic but from distant regions of the world suggest that the genome of enterovirus 70 tends to be conserved during natural infection, a possible consequence of the transient nature of the disease. PMID:6307877

Kew, O M; Nottay, B K; Hatch, M H; Hierholzer, J C; Obijeski, J F

1983-08-01

389

Effect of thymic stromal lymphopoietin on MUC5B expression in human airway epithelial cells.  

PubMed

Thymic stromal lymphopoietin (TSLP) is an interleukin 7-like cytokine and a potent factor for B- and T-cell growth and differentiation. Recent studies have demonstrated an association of TSLP with allergic and inflammatory airway diseases. However, no study on the effect of TSLP on expression of mucin genes in airway epithelial cells has been reported. Therefore, the effects and brief signaling pathways of TSLP on expression of mucin genes in human airway epithelial cells were investigated in this study. In mucin-producing human NCI-H292 airway epithelial cells and primary cultures of normal nasal epithelial cells, the effect and signaling pathway of TSLP on expression of mucin genes were investigated using reverse transcriptase-polymerase chain reaction (RT-PCR), real-time PCR, enzyme immunoassay, and immunoblot analysis with several specific inhibitors and small interfering RNA (siRNA). In human NCI-H292 airway epithelial cells, TSLP increased MUC5B expression. TSLP significantly activated phosphorylation of ERK1/2 and p38 mitogen-activated protein kinase (MAPK). U0126 (ERK1/2 MAPK inhibitor) and SB203580 (p38 MAPK inhibitor) significantly attenuated TSLP-induced MUC5B mRNA expression. Knockdown of ERK1, ERK2, and p38 MAPK by ERK1, ERK2, and p38 MAPK siRNA significantly blocked TSLP-induced MUC5B mRNA expression. In the primary cultures of normal nasal epithelial cells, TSLP significantly increased MUC5B mRNA expression, which was significantly attenuated after pretreatment with U0126 and SB203580. These results suggest that TSLP induces MUC5B expression via the ERK1/2 and p38 MAPK signaling pathway in human airway epithelial cells. PMID:24792379

Bae, Chang Hoon; Choi, Yoon Seok; Song, Si-Youn; Kim, Yong-Dae

2014-05-30

390

MicroRNA-30e* promotes human glioma cell invasiveness in an orthotopic xenotransplantation model by disrupting the NF-?B/I?B? negative feedback loop  

PubMed Central

Constitutive activation of NF-?B is a frequent event in human cancers, playing important roles in cancer development and progression. In nontransformed cells, NF-?B activation is tightly controlled by I?Bs. I?Bs bind NF-