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Coxsackie A virus-associated herpetiform angina.  


Virological investigations were performed in 25 herpangina cases that occurred in a large urban centre in the summer season of 1982. Twelve Coxsackie A virus strains (10 Coxsackie A5 strains, one Coxsackie A4 and one Coxsackie A6 strain) could be isolated from 9 (36.0%) of the patients. PMID:6324449

Zavate, O; Avram, G; Pavlov, E; Burlea-Iriciuc, A; Ivan, A; Cotor, F



Myocardial imaging. Coxsackie myocarditis  

SciTech Connect

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

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



Antenatal and Postnatal Diagnosis of Coxsackie B4 Infection: Case Series  

PubMed Central

Enteroviruses are a common cause of neonatal infection. In particular, Coxsackie B viruses are often associated with severe, fatal disease. The antenatal diagnosis of Coxsackie B viral infections is uncommon. We present a unique case of Coxsackie B4 virus ventriculitis and myocarditis causing fetal hydrops at 22 weeks gestation. Transmission was inferred by viral isolation from the amniotic fluid and by placental pathology. We also describe two additional cases of fatal neonatal Coxsackie B4 infection complicated by myocarditis and encephalitis with cerebral necrosis in a 4-day-old female and by myocarditis, spinal leptomeningitis, and hepatitis in a 4-day-old male. Transplacental acquisition of infection carries a poor prognosis. We propose that Coxsackie B virus should be considered in the investigation of nonimmune hydrops, particularly in the presence of cardiac dysfunction. PMID:23946895

Hunt, Jennifer C.; Schneider, Carol; Menticoglou, Savas; Herath, Jayantha; Del Bigio, Marc R.



Structural Variations in Species B Adenovirus Fibers Impact CD46 Association  

SciTech Connect

A majority of species B adenoviruses (Ads) use CD46 as their primary receptor; however, the precise mechanisms involved in the binding of different Ad types to CD46 have not been resolved. Although previous studies indicate close similarities between two members of species B2 Ads in their usage of CD46, our current investigations revealed a surprisingly low CD46 binding affinity of the species B1 Ad16 fiber knob (equilibrium dissociation constant of 437 nM). We determined the crystal structure of the Ad16 fiber knob and constructed a model of this protein in complex with CD46. A comparison of this model to that of the CD46-Ad11 complex revealed structural differences in the FG and IJ loops that are part of the CD46 binding site. An analysis of a panel of recombinant fiber knobs with mutations targeting these regions in Ad16 and Ad11 uncovered a major contribution of the FG loop on CD46 binding. Two extra residues in the FG loop of the Ad16 fiber significantly reduce receptor interaction. Although avidity effects permit the use of CD46 on host cells by Ad16, virus binding occurs with lower efficiency than with B2 Ad types. The longer FG loop of the Ad16 fiber knob also is shared by other species B1 Ad fibers and, thus, may contribute to the low CD46 binding efficiencies observed for these Ad types. Our findings provide a better understanding of how different Ad types associate with CD46 and could aid in the selection of specific Ad fibers for more efficient Ad gene delivery vectors.

Pache, Lars; Venkataraman, Sangita; Reddy, Vijay S.; Nemerow, Glen R. (Scripps)



Acute myocardial infarction spurred by myopericarditis in a young female patient: Coxsackie B2 to blame.  


Myocardial virus infection may mimic but also trigger acute myocardial infarction.The present paper reports an exceedingly rare presentation of an acute myocardial infarction in a very young female associated with Coxsackie B2 virus infection. A 17-year-old woman with no prior medical history presented to the Cardiac Intensive Care unit with chest pain, ST segment elevation and increased cardiac troponin with pericardial effusion one week after experiencing febrile viral gastroenteritis. Given the age and health of the patient, myocarditis was initially presumed. Coronary angiography and cardiac magnetic resonance imaging studies, however, demonstrated an acute posterior myocardial infarction related to a right coronary artery thrombosis. Serological studies disclosed a concurrent Coxsackie B2 virus infection. The patient made a successful recovery with subsequent minor left ventricular dysfunction at long-term follow-up. Coxsackie B2 myocarditis might have triggered a coronary artery spasm and subsequent thrombosis. PMID:21446385

Liapounova, Natalia A; Mouquet, Frédéric; Ennezat, Pierre V



The Dual Impact of Coxsackie and Adenovirus Receptor Expression on Human Prostate Cancer Gene Therapy1  

Microsoft Academic Search

In a recent paper, we reported a significant difference in coxsackie and adenovirus receptor (CAR) from several human bladder cancer cell lines that correlated with their sensitivities to adenoviral infection (Y. Li, R-C. Pong, J. M. Bergelson, M, C. Hall, A. I. Sagalowsky, C-P. Tseng, Z. Wang, and J. T. Hsieh, Cancer Res., 59: 325-330, 1999). In human prostate cancer,

Takatsugu Okegawa; Yingming Li; Rey-Chen Pong; Jeffrey M. Bergelson; Jian Zhou; Jer-Tsong Hsieh


A recent epidemic of Coxsackie virus type A24 acute haemorrhagic conjunctivitis in Singapore.  

PubMed Central

A recent epidemic of acute conjunctivitis in Singapore showed again the importance of Coxsackie virus type A24 variant as a causative agent of acute haemorrhagic conjunctivitis (AHC). Although the ocular manifestations appeared similar to those described for the 1970 and 1975 outbreaks, a markedly higher rate of respiratory involvements was noted. Not observed in previous epidemics were herpes-like vesicles in the conjunctiva and eyelids of one patient and vesicles in the buccal mucosa and lips of another from whom Coxsackie virus A24 was isolated. The most interesting finding in this study was the isolation of five wild (non-Sabin) poliovirus type 1 strains. Three strains were obtained from conjunctival and two from throat swabs of patients with mild to severe conjunctivitis. It is conceivable that the rare reports of polio-like paralysis or radiculomyelitis accompanying or following AHC in a few Asian countries could be attributed to concurrent infections with a poliovirus and either enterovirus type 70 or Coxsackie virus type A24. Images PMID:3024697

Yin-Murphy, M; Baharuddin-Ishak; Phoon, M C; Chow, V T



Atypical hand, foot, and mouth disease: a vesiculobullous eruption caused by Coxsackie virus A6.  


A previously well infant aged 9 months presented with an acute, self-limiting illness characterised by high fever and a papular eruption that started on the face. Although fever subsided within 3 days, the rash worsened and extended over the whole body, with some papules evolving into vesiculobullous lesions. The infant had been exposed to children with a similar illness 1 week before onset. PCR of vesicular swabs and stool samples taken on day 6 of illness showed Coxsackie virus A6. The illness resolved within 10 days of onset, although onychomadesis was seen on both big toes at follow-up 5 weeks later. Our case exemplifies the severe, atypical cases of hand, foot, and mouth disease that have been reported worldwide since 2008, and in the USA since the 2011. Atypical hand, foot, and mouth disease is caused by a new lineage of Coxsackie virus A6 and is characterised by high fever and vesiculobullous eruptions on the calves and backs of the hands. Infants with eczema might be predisposed to severe disease. PMID:24287184

Feder, Henry M; Bennett, Nicholas; Modlin, John F




EPA Science Inventory

The objectives of this research are to improve on the current analytical methods for quantitative detection of infective coxsackie and echo viruses in drinking water. The specific objectives of this research are to: (1) Improve the sensitivity and specificity of IMS-PCR for in...



EPA Science Inventory

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


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

E-print Network

-resistance and may show synergy with chemo- therapy or radiation therapy approaches for advanced cancers[CANCER RESEARCH 63, 847­853, February 15, 2003] Modulation of Coxsackie-Adenovirus Receptor 35294, and Cancer Gene Therapy Group, Rational Drug Development Program, Department of Oncology

Hemminki, Akseli


Study of Coxsackie B viruses interactions with Coxsackie Adenovirus receptor and Decay-Accelerating Factor using Human CaCo-2 cell line  

PubMed Central

Background Decay Accelerating Factor (DAF) and Coxsackievirus-Adenovirus Receptor (CAR) have been identified as cellular receptors for Coxsackie B viruses (CV-B). The aim of this study is to elucidate the different binding properties of CV-B serotypes and to find out if there are any amino acid changes that could be associated to the different phenotypes. Twenty clinical CV-B isolates were tested on CaCo-2 cell line using anti-DAF (BRIC216) and anti-CAR (RmcB) antibodies. CV-B3 Nancy prototype strain and a recombinant strain (Rec, CV-B3/B4) were tested in parallel. The P1 genomic region of 12 CV-B isolates from different serotypes was sequenced and the Trans-Epithelial Electrical Resistance (TEER) along with the virus growth cycle was measured. Results Infectivity assays revealed clear differences between CV-B isolates with regard to their interactions with DAF and CAR. All tested CV-B isolates showed an absolute requirement for CAR but varied in their binding to DAF. We also reported that for some isolates of CV-B, DAF attachment was not adapted. Genetic analysis of the P1 region detected multiple differences in the deduced amino acid sequences. Conclusion Within a given serotype, variations exist in the capacity of virus isolates to bind to specific receptors, and variants with different additional ligands may arise during infection in humans as well as in tissue culture. PMID:24885774



Experimental infection of tree shrews(Tupaia belangeri) with Coxsackie virus A16.  


Coxsackie virus A16 (CA16) is commonly recognized as one of the main human pathogens of hand-foot-mouth disease (HFMD). The clinical manifestations of HFMD include vesicles of hand, foot and mouth in young children and severe inflammatory CNS lesions. In this study, experimentally CA16 infected tree shrews(Tupaia belangeri) were used to investigate CA16 pathogenesis. The results showed that both the body temperature and the percentages of blood neutrophilic granulocytes / monocytes of CA16 infected tree shrews increased at 4-7 days post infection. Dynamic distributions of CA16 in different tissues and stools were found at different infection stages. Moreover, the pathological changes in CNS and other organs were also observed. These findings indicate that tree shrews can be used as a viable animal model to study CA16 infection. PMID:25465084

Li, Jian-Ping; Liao, Yun; Zhang, Ying; Wang, Jing-Jing; Wang, Li-Chun; Feng, Kai; Li, Qi-Han; Liu, Long-Ding




PubMed Central

Following oral administration of Coxsackie viruses (C viruses) to susceptible chimpanzees, these agents can be isolated from the throat for a period of approximately a week, from the blood for a few days, and from the stools for 2 to 3 weeks or even longer. Animals so infected respond with the formation of specific neutralizing antibodies which are maintained for at least 1 to 2 years. Such chimpanzees are immune when challenged orally with homologous strains of virus. They then excrete virus in the stools for 3 or 4 days (passive transfer); no virus can be recovered from the throats and blood of these animals, and neutralizing antibody levels remain unchanged. Animals immune to one antigenic type of C virus can be infected by feeding a different antigenic type. Following such a heterotypic challenge, virus can again be isolated from the throat, blood, and stools; neutralizing antibodies develop to the new strain. Antibodies to the Texas-1 type of C virus were already present in four chimpanzees upon their arrival in the laboratory from Africa. It was possible to set up intestinal carriage of the Texas-1 virus in these animals and to demonstrate a 10- to 100-fold increase in titer of neutralizing, as well as complement-fixing, antibody. Quantitative titrations of the amount of virus present in the stools and throat were performed. Immediately after the first feeding of virus relatively large amounts can be detected in the stools; the titer drops, but may be maintained for as long as 25 days at 10–2 to 10–3, furnishing evidence that virus multiplication has taken place. Virus in the throat reached the same order of magnitude at first as in the stools but there was a rapid decline to zero in a few days. The Ohio-1 virus differed from the others in that it persisted in the throat as long as in the stools, and in several instances reached a higher titer in the throat. Following homotypic challenge with all types, virus could be detected in the stools for only a relatively short period of time and at low concentration. Virus-neutralizing substances could not be detected in the stools or throat swabs of convalescent animals, at a time when their serum-neutralizing antibody titers were high. Under the limited conditions of the present experiments, C viruses had no effect on the infection of chimpanzees with three different antigenic types of poliomyelitis virus. Both poliomyelitis and C viruses set up independent infections without apparent interaction between them. No enhancement of the poliomyelitis infection took place, as was plain from the fact that none of the infected chimpanzees became paralyzed. a titration of Texas-1 C virus in four chimpanzees revealed that a suspension of infected tissue diluted to 106.0 could cause the development of the carrier state; accidental infection of control animals with other Coxsackie types indicated also that very little virus may be necessary to initiate infection. Seven distinct antigenic types of C virus were inoculated subcutaneously and intramuscularly at the same time into four chimpanzees. The response was the same as that following feeding; virus could be recovered from the throat, blood, and stools, and the animals developed neutralizing antibodies to all seven types of C virus. There was no detectable interference among the viruses. Cortisone did not bring about the reappearance of the virus excretion in chimpanzees previously infected and shown to be intestinal carriers of poliomyelitis and Coxsackie viruses. PMID:13052807

Melnick, Joseph L.; Kaplan, Albert S.



Adenovirus-mediated thymidine kinase gene therapy and coxsackie adenovirus receptor expression in ovarian cancer cells.  


Coxsackie adenovirus receptor (CAR) expression is the main mechanism of adenovirus entry into target cells. It is unclear whether CAR expression itself is influenced by transduction with the adenovirus-Rous sarcoma virus-thymidine kinase (ADV-RSV-TK) gene therapy construct or by the subsequent intracellular accumulation of the TK gene product. Antibody generation and characterization, immunocytochemistry, Western blotting and 3-(4,5-dimethylthiazol)-2,5-diphenyl tetrazolium bromide (MTT) assay were performed to investigate the relationship of gene transfer and CAR expression as well as differences in therapeutic susceptibility of MDAH-2774 and OVCAR-3 cell lines to ADV-RSV-TK gene therapy. CAR expression was observed on the membranes but intracellular translocation of CAR also took place dependent on cellular growth patterns. TK gene expression was dependent on multiplicity of infection (MOI) and thus on vector dose in a linear fashion. Neither TK expression nor ADV transduction influenced CAR expression, or ADV-RSV-TK transduction. Differential susceptibility of different cell lines to TK-induced cell killing by acyclovir metabolites was observed. CAR expression appears not to be influenced by adenoviral transduction or by the accumulation of the TK gene product. Differences in therapeutic sensitivity are most likely mediated by intracellular mechanisms and not by modulation of CAR expression. PMID:19287072

Kieback, D G



Expression of coxsackie and adenovirus receptor distinguishes transitional cancer states in therapy-induced cellular senescence  

PubMed Central

Therapy-induced cellular senescence describes the phenomenon of cell cycle arrest that can be invoked in cancer cells in response to chemotherapy. Sustained proliferative arrest is often overcome as a contingent of senescent tumor cells can bypass this cell cycle restriction. The mechanism regulating cell cycle re-entry of senescent cancer cells remains poorly understood. This is the first report of the isolation and characterization of two distinct transitional states in chemotherapy-induced senescent cells that share indistinguishable morphological senescence phenotypes and are functionally classified by their ability to escape cell cycle arrest. It has been observed that cell surface expression of coxsackie and adenovirus receptor (CAR) is downregulated in cancer cells treated with chemotherapy. We show the novel use of surface CAR expression and adenoviral transduction to differentiate senescent states and also show in vivo evidence of CAR downregulation in colorectal cancer patients treated with neoadjuvant chemoradiation. This study suggests that CAR is a candidate biomarker for senescence response to antitumor therapy, and CAR expression can be used to distinguish transitional states in early senescence to study fundamental regulatory events in therapy-induced senescence. PMID:21364674

Wu, P C; Wang, Q; Dong, Z M; Chu, E; Roberson, R S; Ivanova, I C; Wu, D Y



Immunological and Biochemical Characterization of Coxsackie Virus A16 Viral Particles  

PubMed Central

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

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



Coxsackie and Adenovirus Receptor is increased in adipose tissue of obese subjects: A role for adenovirus infection?  


Context: The Coxsackie and Adenovirus Receptor (CAR) was originally identified as a common receptor for coxsackie B viruses and type C adenoviruses Objective: To investigate CAR gene expression in human adipose tissue to explore its associations with adipocyte physiology. Design and Setting: This was an ex vivo study in 91 visceral (VAT) and 109 subcutaneous adipose tissue (SAT) human samples (61 paired) obtained during elective surgical procedures Patients: Patients were recruited at the Endocrinology Service of the Hospital Universitari Dr. Josep Trueta Main outcome measure: CAR mRNA was measured in human adipose tissue samples and confirmed at protein level and in adipose tissue fractions. The effects of inflammatory stimuli on CAR gene expression were also evaluated. Results: In paired samples, CAR was 46-fold higher in VAT vs. SAT (p<0.0001), being higher also at protein level (p = 0.04). CAR was predominantly found in stromal vascular cell fraction (SVF) in both fat depots. CAR mRNA (p=0.006) and protein levels (p=0.01) in VAT were significantly increased in obese vs. non-obese subjects. In fact, CAR mRNA levels in SAT were positively associated with BMI (r=0.26; p=0.008) and percent fat mass (r=0.28; p=0.004). In VAT, MGLL, FSP27, AKAP, omentin, TKT, S14 and FABP contributed independently to CAR mRNA variation after adjusting for age and BMI. Macrophage-conditioned medium led to increased CAR gene expression in mature adipocytes in vitro. Conclusions: The increased expression of CAR in adipose tissue from obese subjects, mainly in SVFs, suggests that CAR might play a role in adipose tissue dysfunction given its dual associations with adipogenic and inflammatory genes. PMID:25459915

Serrano, Marta; Moreno, María; Bassols, Judit; Moreno-Navarrete, José María; Ortega, Francisco; Ricart, Wifredo; Manuel Fernández-Real, José



Trace element distribution in heart tissue sections studied by nuclear microscopy is changed in coxsackie virus B3 myocarditis in methyl mercury-exposed mice  

Microsoft Academic Search

Methyl mercury (MeHg) has been shown to change Coxsackie virus type B3 (CB3) myocarditis in a direction compatible with the\\u000a development of chronic disease. Murine models of CB3 myocarditis closely mimic the pathogenesis in humans. There are also\\u000a indications that metals, such as mercury, and trace elements may interact and adversely affect viral replication and development\\u000a of inflammatory lesions. The

Nils-Gunnar Ilbäck; Ulf Lindh; Lars Wesslén; Jan Fohlman; Göran Friman



[An experimental model of the vertical transmission of the Coxsackie-group enteroviruses and its use in developing methods to prevent congenital coxsackievirus infection].  


An experimental model of vertical transmission of Coxsackie group enteroviruses was developed in BALB/c mice the first generation of which was infected with Coxsackie A18 virus in the neonatal period. Persistence of the virus was demonstrated in all females of the first generation tested and in 90.9% to 100% of the animals of the next two generations. Cytochemical analysis of the enzyme status of lymphocytes revealed reliable relationship between the depression of energetic metabolism enzymes and the activity of virus amplification in the animals under study. The correction of aerobic respiration in pregnant females by administration of a complex of energy metabolism substrates and cofactors was accompanied by a significant reduction of the virus infection activity in the females and their offsprings as well as by prevention of transplacental infection in some litters. The experimental model of vertical transmission of enteroviruses is proposed for use in the development of methods for prevention of congenital Coxsackie virus infection. The authors express their deep gratitude to the sponsor of the publication--NOVRUZ Co., Turkmenistan. PMID:8284921

Lozovskaia, L S; Zubkova, I V; Khellenov, E A; Shishchenko, V M; Konopleva, T N



A rapid assay for evaluation of antiviral activity against coxsackie virus B3, influenza virus A, and herpes simplex virus type 1.  


In order to identify new potential antiviral drugs, small amounts of extracts or compounds have to be examined for cytotoxicity and antiviral activity in primary screening using a rapid, easy, inexpensive, and highly standardised test system. In this study, high-throughput cytopathic effect (CPE) inhibitory assays were established for coxsackie virus B3 on HeLa Ohio cells, influenza virus A on Madin-Darby canine kidney cells, and herpes simplex virus type 1 (HSV-1) on green monkey kidney cells that meet these requirements. The cytotoxic and the antiviral effects were quantified using a crystal violet uptake assay allowing automated handling of large numbers of candidate agents. To ensure comparable results with plaque reduction assays, the 50 and 90% plaque inhibitory concentrations of guanidine, amantadine, and phosphonoformic acid were used to standardise the anti-coxsackie virus B3, anti-influenza virus A, and anti-HSV-1 tests, respectively. The strong correlation between the antiviral activity determined by CPE-inhibitory assays and plaque reduction assay was further proved for other antivirals. In summary, low amounts of large numbers of compounds may be tested inexpensively and standardised within 24 h (coxsackie virus B3 and influenza virus A) or 48 h (herpes simplex virus type 1) post-infection using CPE inhibitory assays. PMID:11377720

Schmidtke, M; Schnittler, U; Jahn, B; Dahse, H; Stelzner, A



Adenovirus tumor targeting and hepatic untargeting by a coxsackie/adenovirus receptor ectodomain anti-carcinoembryonic antigen bispecific adapter.  


Adenovirus vectors have a number of advantages for gene therapy. However, because of their lack of tumor tropism and their preference for liver infection following systemic administration, they cannot be used for systemic attack on metastatic disease. Many epithelial tumors (e.g., colon, lung, and breast) express carcinoembryonic antigen (CEA). To block the natural hepatic tropism of adenovirus and to "retarget" the virus to CEA-expressing tumors, we used a bispecific adapter protein (sCAR-MFE), which fuses the ectodomain of the coxsackie/adenovirus receptor (sCAR) with a single-chain anti-CEA antibody (MFE-23). sCAR-MFE untargets adenovirus-directed luciferase transgene expression in the liver by >90% following systemic vector administration. Moreover, sCAR-MFE can "retarget" adenovirus to CEA-positive epithelial tumor cells in cell culture, in s.c. tumor grafts, and in hepatic tumor grafts. The sCAR-MFE bispecific adapter should, therefore, be a powerful agent to retarget adenovirus vectors to epithelial tumor metastases. PMID:17545616

Li, Hua-Jung; Everts, Maaike; Pereboeva, Larisa; Komarova, Svetlana; Idan, Anat; Curiel, David T; Herschman, Harvey R



Antiviral Activity of Total Flavonoid Extracts from Selaginella moellendorffii Hieron against Coxsackie Virus B3 In Vitro and In Vivo  

PubMed Central

The antiviral activity of total flavonoid extracts from Selaginella moellendorffii Hieron and its main constituents amentoflavone were investigated against coxsackie virus B3 (CVB3). When added during or after viral infection, the extracts and amentoflavone prevented the cytopathic effect (CPE) of CVB3, as demonstrated in a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) colorimetric assay, with a 50% inhibitory concentration (IC50) from 19 ± 1.6 to 41 ± 1.2??g/mL and 25 ± 1.2 to 52 ± 0.8??g/mL, respectively. KM mice were used as animal models to test the extracts' activity in vivo. Oral administration of the total flavonoid extracts at 300?mg/kg/day significantly reduced mean viral titers in the heart and kidneys as well as mortality after infection for 15 days. The experimental results demonstrate that in vitro and in vivo the model mice infected with CVB3 can be effectively treated by the total flavonoid extracts from Selaginella moellendorffii Hieron. PMID:24963331

Yin, Dan; Li, Juan; Lei, Xiang; Liu, Yimei; Yang, Zhanqiu; Chen, Keli



Antiviral activity of arbidol against influenza A virus, respiratory syncytial virus, rhinovirus, coxsackie virus and adenovirus in vitro and in vivo  

Microsoft Academic Search

Summary  Arbidol, ethyl-6-bromo-4-[(dimethylamino)-methyl]-5-hydroxy-1-methyl-2-[(phenylthio)methyl]-in dole-3-carboxylate hydrochloride\\u000a monohydrate, is an antiviral chemical agent. In this report, we studied the antiviral activity of arbidol against a panel\\u000a of human respiratory viruses, namely influenza A virus (FLU-A, A\\/PR\\/8\\/34 H1N1), respiratory syncytial virus (RSV), human rhinovirus\\u000a type 14 (HRV 14), coxsackie virus B3 (CVB3) and adenovirus type 7 (AdV-7) in vitro in cell culture. Arbidol was

L. Shi; H. Xiong; J. He; H. Deng; Q. Li; Q. Zhong; W. Hou; L. Cheng; H. Xiao; Z. Yang



Enhanced therapeutic efficacy of an adenovirus-PEI-bile-acid complex in tumors with low coxsackie and adenovirus receptor expression.  


Adenovirus (Ad) is a potential vehicle for cancer gene therapy. However, cells that express low levels of the coxsackie and adenovirus receptor (CAR) demonstrate poor Ad infection efficiency. We developed a bile acid-conjugated poly(ethyleneimine) (DA3)-coated Ad complex (Ad/DA3) to enhance Ad transduction efficiency. The size distribution and zeta potential of Ad/DA3 increased to 324 ± 3.08 nm and 10.13 ± 0.21 mV, respectively, compared with those of naked Ad (108 ± 2.26 nm and -17.7 ± 1.5 mV). The transduction efficiency of Ad/DA3 increased in a DA3 polymer concentration-dependent manner. Enhanced gene transfer by Ad/DA3 was more evident in CAR-moderate and CAR-negative cancer cells. Competition assays with a CAR-specific antibody revealed that internalization of Ad/DA3 was not mediated primarily by CAR but involved clathrin-, caveolae-, and macropinocytosis-mediated endocytosis. Cancer cell death was significantly increased when oncolytic Ad and DA3 were complexed (RdB-KOX/DA3) compared to that of naked oncolytic Ad and was inversely proportional to CAR levels. Importantly, RdB-KOX/DA3 significantly enhanced apoptosis, reduced angiogenesis, reduced proliferation, and increased active viral replication in human tumor xenografts compared to that of naked Ad. These results demonstrate that a hybrid vector system can increase the efficacy of oncolytic Ad virotherapy, particularly in CAR-limited tumors. PMID:24731708

Lee, Cho-Hee; Kasala, Dayananda; Na, Youjin; Lee, Min Sang; Kim, Sung Wan; Jeong, Ji Hoon; Yun, Chae-Ok



Recombination between Poliovirus and Coxsackie A Viruses of Species C: A Model of Viral Genetic Plasticity and Emergence  

PubMed Central

Genetic recombination in RNA viruses was discovered many years ago for poliovirus (PV), an enterovirus of the Picornaviridae family, and studied using PV or other picornaviruses as models. Recently, recombination was shown to be a general phenomenon between different types of enteroviruses of the same species. In particular, the interest for this mechanism of genetic plasticity was renewed with the emergence of pathogenic recombinant circulating vaccine-derived polioviruses (cVDPVs), which were implicated in poliomyelitis outbreaks in several regions of the world with insufficient vaccination coverage. Most of these cVDPVs had mosaic genomes constituted of mutated poliovaccine capsid sequences and part or all of the non-structural sequences from other human enteroviruses of species C (HEV-C), in particular coxsackie A viruses. A study in Madagascar showed that recombinant cVDPVs had been co-circulating in a small population of children with many different HEV-C types. This viral ecosystem showed a surprising and extensive biodiversity associated to several types and recombinant genotypes, indicating that intertypic genetic recombination was not only a mechanism of evolution for HEV-C, but an usual mode of genetic plasticity shaping viral diversity. Results suggested that recombination may be, in conjunction with mutations, implicated in the phenotypic diversity of enterovirus strains and in the emergence of new pathogenic strains. Nevertheless, little is known about the rules and mechanisms which govern genetic exchanges between HEV-C types, as well as about the importance of intertypic recombination in generating phenotypic variation. This review summarizes our current knowledge of the mechanisms of evolution of PV, in particular recombination events leading to the emergence of recombinant cVDPVs. PMID:21994791

Combelas, Nicolas; Holmblat, Barbara; Joffret, Marie-Line; Colbčre-Garapin, Florence; Delpeyroux, Francis



Crosstalk between desmoglein-2/desmocollin-2/Src kinase and coxsackie and adenovirus receptor/ZO-1 protein complexes regulates blood-testis barrier dynamics  

PubMed Central

Morphological studies in the testis reported the presence of ‘desmosome-like’ junctions between Sertoli cells at the blood-testis barrier, whose function is also constituted by tight junctions and basal ectoplasmic specializations. Unfortunately, little is known about the role of desmosomes in blood-testis barrier dynamics. This study aims to fill this gap with the functional investigation of two desmosomal cadherins, desmoglein-2 and desmocollin-2, by their specific knockdown in Sertoli cells cultured in vitro. Reminiscent of the blood-testis barrier in vivo, desmosome-like structures were visible by electron microscopy when Sertoli cells were cultured at high density, thereby forming a polarized epithelium with functional cell junctions. At this point, we opted to focus our efforts on desmoglein-2 and desmocollin-2 based on results which illustrated desmosomal mRNAs to be expressed by Sertoli and germ cells, as well as on results which illustrated desmoglein-2 to co-immunoprecipitate with plakoglobin, c-Src and desmocollin-2. Simultaneous knockdown of desmoglein-2 and desmocollin-2 not only led to a reduction and mislocalization of zonula occludens-1, but also perturbed the localization of c-Src and coxsackie and adenovirus receptor at the cell-cell interface, resulting in disruption of tight junction permeability barrier. We hereby propose a novel regulatory protein complex composed of desmoglein-2, desmocollin-2, c-Src, coxsackie and adenovirus receptor and ZO-1 at the blood-testis barrier. PMID:20188849

Lie, Pearl P.Y.; Cheng, C. Yan; Mruk, Dolores D.



Deletion of the intracellular domain of coxsackie and adenovirus receptor (CAR) enhances the expression of itself and boosts the efficiency of current adenovirus-mediated gene therapy in ovarian cancer cell lines in vitro  

Microsoft Academic Search

The failure of adenovirus-mediated gene therapy often derives from the absence of coxsackie and adenovirus receptor (CAR) expression in target cells. We hypothesize that the slight up-regulation of CAR expression might boost the effect of adenovirus-mediated gene therapy in ovarian cancer. To test this hypothesis, we transfected full-length and intracellular-domain-deleted (tailless) CAR plasmids into CAR-deficient ovarian cancer cell line SKOV3.

Beibei Wang; Gang Chen; Jianfeng Zhou; Peng Wu; Danfeng Luo; Xiaoyuan Huang; Tao Zhu; Zhiqiang Han; Gang Xu; Shixuan Wang; Yunping Lu; Ding Ma



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

SciTech Connect

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

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



The transduction of Coxsackie and Adenovirus Receptor-negative cells and protection against neutralizing antibodies by HPMA-co-oligolysine copolymer-coated adenovirus.  


Adenoviral (AdV) gene vectors offer efficient nucleic acid transfer into both dividing and non-dividing cells. However issues such as vector immunogenicity, toxicity and restricted transduction to receptor-expressing cells have prevented broad clinical translation of these constructs. To address this issue, engineered AdV have been prepared by both genetic and chemical manipulation. In this work, a polymer-coated Ad5 formulation is optimized by evaluating a series of N-(2-hydroxypropyl) methacrylamide (HPMA)-co-oligolysine copolymers synthesized by living polymerization techniques. This synthesis approach was used to generate highly controlled and well-defined polymers with varying peptide length (K(5), K(10) and K(15)), polymer molecular weight, and degradability to coat the viral capsid. The optimal formulation was not affected by the presence of serum during transduction and significantly increased Ad5 transduction of several cell types that lack the Coxsackie and Adenovirus Receptor (CAR) by up to 6-fold compared to unmodified AdV. Polymer-coated Ad5 also retained high transduction capability in the presence of Ad5 neutralizing antibodies. The critical role of heparan sulfate proteoglycans (HSPGs) in mediating cell binding and internalization of polymer-coated AdV was also demonstrated by evaluating transduction in HSPG-defective recombinant CHO cells. The formulations developed here are attractive vectors for ex vivo gene transfer in applications such as cell therapy. In addition, this platform for adenoviral modification allows for facile introduction of alternative targeting ligands. PMID:21959008

Wang, Chung-Huei K; Chan, Leslie W; Johnson, Russell N; Chu, David S H; Shi, Julie; Schellinger, Joan G; Lieber, André; Pun, Suzie H



GYY4137, a hydrogen sulfide?releasing molecule, inhibits the inflammatory response by suppressing the activation of nuclear factor?kappa B and mitogen?activated protein kinases in Coxsackie virus B3?infected rat cardiomyocytes.  


GYY4137 is a water?soluble, small molecule hydrogen sulfide (H2S)?release agent that possesses potent cardioprotective and anti?inflammatory properties in experimental models. Coxsackie virus B3 (CVB3) infection commonly causes viral myocarditis, which mainly involves immune cell infiltration, eventually resulting in heart failure. In the present study, the effects and underlying mechanisms of GYY4137 treatment of CVB3?induced myocarditis were investigated. The effects of GYY4137 on CVB3?induced nuclear factor?kappa B (NF??B) activity were examined by western blotting, immunofluorescence and electrophoretic mobility shift assay. Mitogen?activated protein kinase (MAPK) signaling protein expression levels were detected by western blotting. Cardiomyocyte damage?related enzyme activities, such as lactate dehydrogenase (LDH) and creatine kinase MB (CK?MB), were measured by ELISA, as well as the production of proinflammatory cytokines. The results revealed that GYY4137 suppressed CVB3?induced secretion of LDH, CK?MB and pro?inflammatory cytokines, such as tumor necrosis factor??, interleukin (IL)?1? and IL?6. Furthermore, the activation of NF??B and the I?B? degradation induced by CVB3 were also inhibited by GYY4137. Notably, the phosphorylation of p38, ERK1/2 and JNK1/2 induced by CVB3 was also inhibited by GYY4137. In conclusion, the data demonstrate that GYY4137 exerts anti?inflammatory effects in CVB3?infected cardiomyocytes. This anti?inflammatory mechanism may be associated with suppression of NF??B and MAPK signaling pathway activation. PMID:25377925

Wu, Zubo; Peng, Hua; Du, Qing; Lin, Wen; Liu, Yali



Characterization of a 100-kilodalton binding protein for the six serotypes of coxsackie B viruses.  

PubMed Central

Viral infection of host cells primarily depends on binding of the virus to a specific cell surface protein. In order to characterize the binding protein for group B coxsackieviruses (CVB), detergent-solubilized membrane proteins of different cell lines were tested in virus overlay protein-binding assays. A prominent virus-binding protein with a molecular mass of 100 kDa was detected in various CVB-permissive human and monkey cell lines but was not detected in nonpermissive cell lines. The specificity of CVB binding to the 100-kDa protein on permissive human cells was substantiated by binding of all six serotypes of CVB and by competition experiments. In contrast, poliovirus and Sendai virus did not bind to the 100-kDa CVB-specific protein. A fraction of HeLa membrane proteins enriched in the range of 100 kDa showed functional activity by transforming infectious CVB (160S) into A-particles (135S). In order to purify this CVB-binding protein, solubilized membrane proteins from HeLa cells were separated by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by elution of the 100-kDa protein. Amino acid sequence analysis of tryptic fragments of the CVB-binding protein indicated that this 100-kDa CVB-specific protein is a cell surface protein related to nucleolin. These results were confirmed by immunoprecipitations of the CVB-binding protein with nucleolin-specific antibodies, suggesting that a nucleolin-related membrane protein acts as a specific binding protein for the six serotypes of CVB. PMID:7474086

de Verdugo, U R; Selinka, H C; Huber, M; Kramer, B; Kellermann, J; Hofschneider, P H; Kandolf, R



Recombination between Polioviruses and Co-Circulating Coxsackie A Viruses: Role in the Emergence of Pathogenic  

E-print Network

outbreaks of poliomyelitis caused by pathogenic circulating vaccine-derived polioviruses (cVDPVs) have, poliomyelitis caused by wild polioviruses has disappeared from large parts of the world. However, poliomyelitis

Paris-Sud XI, Université de


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

Microsoft Academic Search

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




A zebrafish coxsackievirus and adenovirus receptor homologue interacts with coxsackie B virus and adenovirus.  


In this study, a zebrafish homologue of the coxsackievirus and adenovirus receptor (CAR) protein was identified. Although the extracellular domain of zebrafish CAR (zCAR) is less than 50% identical to that of human CAR (hCAR), zCAR mediated infection of transfected cells by both adenovirus type 5 and coxsackievirus B3. CAR residues interacting deep within the coxsackievirus canyon are highly conserved in zCAR and hCAR, which is consistent with the idea that receptor contacts within the canyon are responsible for coxsackievirus attachment. In contrast, CAR residues contacting the south edge of the canyon are not conserved, suggesting that receptor interaction with the viral "puff region" is not essential for attachment. PMID:12239327

Petrella, JenniElizabeth; Cohen, Christopher J; Gaetz, Jedidiah; Bergelson, Jeffrey M



Expression of Coxsackie adenovirus receptor and alphav-integrin does not correlate with adenovector targeting in vivo indicating anatomical vector barriers  

Microsoft Academic Search

Recombinant adenoviral vectors are broadly applied in gene therapy protocols. However, adenovector-mediated gene transfer has limitations in vivo. One of these is the low gene transfer rate into organs other than the liver after systemic intravenous vector injection. Local direct injection into the target organ has been used as one possible solution, but increases necessary equipment and methodology and is

H Fechner; A Haack; H Wang; X Wang; K Eizema; M Pauschinger; R G Schoemaker; R van Veghel; A B Houtsmuller; H-P Schultheiss; J M J Lamers; W Poller



ORIGINAL ARTICLE Occurrence of adenovirus and other enteric viruses in  

E-print Network

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

Illinois at Chicago, University of


1023. Adenoviruses Transport across a Polarized Monolayer of Caco-2 Cells by Transferrin Receptor-Mediated Transcytosis  

Microsoft Academic Search

Adenoviral (Ad) vectors are commonly used in gene delivery because they have the capacity to infect a broad range of different cell types. Recent studies have identified a 46-kDa host cell membrane protein, coxsackie-adenovirus receptor (CAR), which serves as the primary receptor for Ad serotypes 2 and 5 as well as coxsackie B virus attachment to cells. Following binding to

Zeng B. Zhu; Sharmila K. Makhija; Baogen Lu; Minghui Wang; Meredith A. Preuss; Angel A. Rivera; Fen Zhou; Gene P. Siegal; Ronald D. Alvarez; David T. Curiel



Altering the Ad5 packaging domain affects the maturation of the Ad particles.  


We have previously described a new family of mutant adenoviruses carrying different combinations of attB/attP sequences from bacteriophage PhiC31 flanking the Ad5 packaging domain. These novel helper viruses have a significantly delayed viral life cycle and a severe packaging impairment, regardless of the presence of PhiC31 recombinase. Their infectious viral titers are significantly lower (100-1000 fold) than those of control adenovirus at 36 hours post-infection, but allow for efficient packaging of helper-dependent adenovirus. In the present work, we have analyzed which steps of the adenovirus life cycle are altered in attB-helper adenoviruses and investigated whether these viruses can provide the necessary viral proteins in trans. The entry of attB-adenoviral genomes into the cell nucleus early at early timepoints post-infection was not impaired and viral protein expression levels were found to be similar to those of control adenovirus. However, electron microscopy and capsid protein composition analyses revealed that attB-adenoviruses remain at an intermediate state of maturation 36 hours post-infection in comparison to control adenovirus which were fully mature and infective at this time point. Therefore, an additional 20-24 hours were found to be required for the appearance of mature attB-adenovirus. Interestingly, attB-adenovirus assembly and infectivity was restored by inserting a second packaging signal close to the right-end ITR, thus discarding the possibility that the attB-adenovirus genome was retained in a nuclear compartment deleterious for virus assembly. The present study may have substantive implications for helper-dependent adenovirus technology since helper attB-adenovirus allows for preferential packaging of helper-dependent adenovirus genomes. PMID:21611162

Alba, Raul; Cots, Dan; Ostapchuk, Philomena; Bosch, Assumpcio; Hearing, Patrick; Chillon, Miguel



Realms of the Viruses Online  

ERIC Educational Resources Information Center

Viruses have evolved strategies for infecting all taxa, but most viruses are highly specific about their cellular host. In humans, viruses cause diverse diseases, from chronic but benign warts, to acute and deadly hemorrhagic fever. Viruses have entertaining names like Zucchini Yellow Mosaic, Semliki Forest, Coxsackie, and the original terminator,…

Liu, Dennis



CAR is a cell–cell adhesion protein in human cancer cells and is expressionally modulated by dexamethasone, TNF?, and TGF?  

Microsoft Academic Search

The coxsackie adenovirus receptor (CAR) has become of interest for gene therapy due to its crucial function in adenoviral cell entry. In clinical trials with adenoviral vectors, dexamethasone is applied to reduce side effects such as inflammatory reactions or emesis. By using a ?-galactosidase-expressing adenovirus (AdGal), we observed that dexamethasone treatment resulted in decreased adenoviral gene transfer into human cancer

A Brüning; I B Runnebaum



Loss of Adenoviral Receptor Expression in Human Bladder Cancer Cells: A Potential Impact on the Efficacy of Gene Therapy1  

Microsoft Academic Search

There is great interest in the development of gene therapeutic strategies for the treatment of benign and malignant diseases. Recombinant adeno- virus has a wide spectrum of tissue specificity and is an efficient vector delivery system. Successful gene delivery, however, requires viral entry into the target cells via specific receptor-mediated uptake. Recently, a cDNA clone (the coxsackie and adenovirus receptor

Yingming Li; Rey-Chen Pong; Jeffrey M. Bergelson; M. Craig; Arthur I. Sagalowsky; Ching-Ping Tseng; Zhi Wang; Jer-Tsong Hsieh



2003 Participating Schools Academy of the Holy Names  

E-print Network

-Kate Lenihan Hilary Lewis Zoe Linder-Baptie Jaimone Lyles Daniel Plaat Akira Rowell Adam Siegel Gregory Kaitlinn Truesell Paul Ziter Cobleskill-Richmondville Middle School Teachers: Mr. Richard Clements and Mrs Morris Lindsay Norman Megan O'Brien Sarah Phelan Steve Pinchook Katie Sutliff Coxsackie-Athens Elementary



EPA Science Inventory

A quantitative enzyme-linked immunosorbent assay (ELISA) was used for identification of viruses selected as representative water-borne viruses: poliovirus 1, echovirus 6, coxsackievirus A9, and coxsackie B viruses. Partially purified viral antigens or virus-specific antibodies we...



EPA Science Inventory

The inactivation kinetics of ClO2 on two enteroviruses, poliovirus 1 (Mahoney) and coxsackie virus A9, and an enteric indicator of fecal pollution, Escherichia coli, were examined in laboratory studies. In addition, the disinfecting ability of ClO2 as affected by particulates (bo...


Viral myocarditis and coagulopathy: Increased tissue factor expression and plasma thrombogenicity  

Microsoft Academic Search

We investigated the effects of viral infection on Tissue Factor (TF) expression and activity in mice within the myocardium to understand increased thrombosis during myocarditis. Mice were infected with coxsackie virus B3 (CVB3) and the hearts were collected at day 4, 8 and 28 post infection (p.i.). Myocardial TF expression and cellular activity as well as plasma activity were analyzed

Silvio Antoniak; Ulrike Boltzen; Alexander Riad; Angela Kallwellis-Opara; Maria Rohde; Andrea Dörner; Carsten Tschöpe; Michel Noutsias; Matthias Pauschinger; Heinz-Peter Schultheiss; Ursula Rauch



[The determination of the antiviral activity of the sorbent Evirkhip (in vitro studies)].  


It was found that "Evirkhip" possesses a marked antiviral activity confirmed on a model of enteroviruses (coxsackie). The authors also studied the resistance to the drug in the chain: eukaryotic cell-viral particle-prokaryotic cell. The resistance decreased with the shift to right. PMID:1336633

Bo?ko, I I; Rudychenko, V F; Volkova, V P; Safro, H P; Bondarenko, L I



Susceptibility of the VERO line of African green monkey kidney cells to human enteroviruses  

PubMed Central

The relative susceptibility of VERO cells and primary rhesus monkey kidney cells to 47 prototype strains of human enteroviruses is described. Of these strains, types 4, 14, 16, 17, 18, 21, 31 and 34 and Coxsackie virus A 9 failed to cause CPE in the VERO cells whilst only one, echovirus type 34, failed to cause CPE in the monkey kidney cells. A comparison is given of the efficiency of the two cell cultures for enterovirus isolation from clinical material. Results show that VERO cells are as useful as primary monkey kidney for the isolation of Coxsackie B viruses but less satisfactory for isolating echoviruses. They are satisfactory for the isolation of single types of poliovirus and appear to be more satisfactory than primary monkey kidney cells for the isolation of mixtures of polioviruses. The identification of enteroviruses by neutralization tests in VERO cells is successful. PMID:4361500

Davis, Patricia M.; Phillpotts, R. J.



N-hexanoyl chitosan-stabilized magnetic nanoparticles: enhancement of adenoviral-mediated gene expression both in vitro and in vivo  

Microsoft Academic Search

We developed hexanoyl chloride–modified chitosan (Nac-6) stabilized iron oxide nanoparticles (Nac-6-IOPs) as magnetic nanoparticles for viral gene (Ad\\/LacZ) delivery via magnetofection. This vector, Nac-6-IOPs\\/Ad\\/LacZ, binds to K562 cells in the presence of external magnetic fields and results in enhanced expression of the transgene in those cells that do not exhibit the coxsackie-adenovirus receptor (CAR). Our results demonstrate that Nac-6-IOPs\\/Ad\\/LacZ is

Shanta Raj Bhattarai; Sun Young Kim; Kyu Yun Jang; Ki Chang Lee; Ho Keun Yi; Dae Yeol Lee; Hak Yong Kim; Pyoung Han Hwang



Collection and Testing of Respiratory Samples

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



An Advanced Generation of Adenoviral Vectors Selectively Enhances Gene Transfer for Ovarian Cancer Gene Therapy Approaches  

Microsoft Academic Search

Objective. We hypothesized that incorporation of an integrin binding Arg-Gly-Asp (RGD)-containing peptide to the HI loop of the adenovirus fiber knob would allow enhanced, coxsackie-adenovirus receptor-independent gene transfer by modified Ad vector in the context of ovarian cancer.Methods. Ovarian cancer cell lines, primary ovarian cancer cells, primary tumor explants, and mesothelial tissue were transfected with luciferase encoding adenovirus (AdCMVLuc) or

Timothy J. Vanderkwaak; Minghui Wang; Jesús Gómez-Navarro; Claudine Rancourt; Igor Dmitriev; Victor Krasnykh; Mack Barnes; Gene P. Siegal; Ronald Alvarez; David T. Curiel



Preclinical evaluation of a class of infectivity-enhanced adenoviral vectors in ovarian cancer gene therapy  

Microsoft Academic Search

Ovarian carcinoma cells are often infected inefficiently by adenoviruses (Ad) due to low expression of coxsackie–adenovirus receptors (CAR), hindering the application of adenovirus-mediated gene therapy in ovarian cancer. In this study, we explored a class of infectivity-enhanced Ad vectors, which contain CAR-independent targeting motifs RGD (Ad5.RGD), polylysine (Ad5.pK7), or both (Ad5.RGD.pK7), for their utility in ovarian cancer gene therapy using

H Wu; T Han; J T Lam; C A Leath; I Dmitriev; E Kashentseva; M N Barnes; R D Alvarez; D T Curiel; DT Curiel



Construction of Urothelium-Specific Recombinant Adenovirus and Its Inhibition in Bladder Cancer Cell  

Microsoft Academic Search

Aim: To construct urothelium-specific recombinant adenovirus and investigate its inhibition in bladder cancer cell. Methods: RT-PCR analysis was used to determine expression patterns of hUPII and coxsackie adenovirus receptor on multiple cell lines. Transient transfection and luciferase detecting assay were used to detect tissue specificity of the hUPII promoter. Recombinant adenovirus Ad-UPII-E1A and Ad-UPII-Null were constructed. Restrictive enzyme digestion assay

Xiang-Dong He; Zhi-Ping Wang; Hai-Yan Wei; Qin Zhou; De-Gui Wang; Jun-Qiang Tian; Sheng-Jun Fu; Ronald Rodriguez



Antiviral activity of galangin isolated from the aerial parts of Helichrysum aureonitens  

Microsoft Academic Search

The in vitro antiviral activity of galangin (3,5,7-trihydroxyflavone), the major antimicrobial compound isolated from the shoots of Helichrysum aureonitens, was investigated against herpes simplex virus type 1 (HSV-1), coxsackie B virus type 1 (Cox B1), adenovirus type 31 (Ad31) and reovirus. At concentrations ranging from 12–47 ?g\\/ml galangin showed significant antiviral activity against HSV-1 and CoxB1, limited activity against reovirus,

J. J. M. Meyer; A. J. Afolayan; M. B. Taylor; D. Erasmus



Preclinical Characterization of the Antiglioma Activity of a Tropism-Enhanced Adenovirus Targeted to the Retinoblastoma Pathway  

Microsoft Academic Search

Background: Oncolytic adenoviruses are promising thera- pies for the treatment of gliomas. However, untargeted viral replication and the paucity of coxsackie-adenovirus recep- tors (CARs) on tumor cells are major stumbling blocks for adenovirus-based treatment. We studied the antiglioma ac- tivity of the tumor-selective Delta-24 adenovirus, which en- compasses an early 1 A adenoviral (E1A) deletion in the retinoblastoma (Rb) protein-binding

Juan Fueyo; Ramon Alemany; Candelaria Gomez-Manzano; Gregory N. Fuller; Asadullah Khan; Charles A. Conrad; Ta-Jen Liu; Hong Jiang; Michael G. Lemoine; Kaori Suzuki; Raymond Sawaya; David T. Curiel; W. K. Alfred Yung; Frederick F. Lang



Treatment of Ovarian Cancer with a Tropism Modified Oncolytic Adenovirus1  

Microsoft Academic Search

Ad5-24RGD is an adenovirus that is selectively replication competent in cells defective in the Rb\\/p16 pathway, such as ovarian cancer cells. The fiber of Ad5-24RGD contains an integrin binding RGD-4C motif, allow- ing Coxsackie adenovirus receptor-independent infection of cancer cells. Oncolysis of cell lines was similar to that of a wild-type control, and replication in primary tumor material was shown

Gerd J. Bauerschmitz; John T. Lam; Anna Kanerva; Kaori Suzuki; Dirk M. Nettelbeck; Igor Dmitriev; Victor Krasnykh; Galina V. Mikheeva; Mack N. Barnes; Ronald D. Alvarez; Peter Dall; Ramon Alemany; David T. Curiel; Akseli Hemminki



Identification of a series of compounds with potent antiviral activity for the treatment of enterovirus infections.  


Rhinovirus (genus enterovirus) infections are responsible for many of the severe exacerbations of asthma and chronic obstructive pulmonary disease. Other members of the genus can cause life-threatening acute neurological infections. There is currently no antiviral drug approved for the treatment of such infections. We have identified a series of potent, broad-spectrum antiviral compounds that inhibit the replication of the human rhinovirus, Coxsackie virus, poliovirus, and enterovirus-71. The mechanism of action of the compounds has been established as inhibition of a lipid kinase, PI4KIII?. Inhibition of hepatitis C replication in a replicon assay correlated with enterovirus inhibition. PMID:24900715

MacLeod, Angus M; Mitchell, Dale R; Palmer, Nicholas J; Van de Poël, Hervé; Conrath, Katja; Andrews, Martin; Leyssen, Pieter; Neyts, Johan



Microorganisms in the aetiology of atherosclerosis  

PubMed Central

Recent publications have suggested that infective pathogens might play an important role in the pathogenesis of atherosclerosis. This review focuses on these microorganisms in the process of atherosclerosis. The results of in vitro studies, animal studies, tissue studies, and serological studies will be summarised, followed by an overall conclusion concerning the strength of the association of the microorganism with the pathogenesis of atherosclerosis. The role of the bacteria Chlamydia pneumoniae and Helicobacter pylori, and the viruses human immunodeficiency virus, coxsackie B virus, cytomegalovirus, Epstein-Barr virus, herpes simplex virus, and measles virus will be discussed. Key Words: atherosclerosis • Chlamydia pneumoniae • Helicobacter pylori PMID:11041053

Morre, S; Stooker, W; Lagrand, W; van den Brule, A J C; Niessen, H



Transplacental infection of Coxsackievirus B3 pathological findings in the fetus.  


Coxsackievirus intrauterine infection has been documented mostly on the basis of indirect evidence of transplacental transmission, with neonatal manifestations ranging from asymptomatic infection to meningoencephalitis, myocarditis, and generalized sepsis. This is the first report of prenatal findings and fetoplacental pathology in a third trimester fetus with coxsackie B3 transplacental infection confirmed by molecular techniques. Prenatal ultrasound detected severe reduction of fetal movements at the 27th week. Late onset fetal akinesia deformation sequence with mild arthrogryposis, necrotic meningoencephalitis with vascular calcifications, interstitial pneumonitis, mild myocardial hypertrophy, and chronic monocytic placental villitis were the cardinal findings at fetal autopsy following interruption of the pregnancy. PMID:17457913

Konstantinidou, Anastasia; Anninos, Hector; Spanakis, Nikolaos; Kotsiakis, Xenophon; Syridou, Garyfallia; Tsakris, Athanassios; Patsouris, Efstratios



Hand, foot, and mouth syndrome in an immunocompetent adult: a case report  

PubMed Central

Background Hand, foot, and mouth syndrome (HFMS) is a common acute illness. It is characterized by mild clinical symptoms including fever, blisters, and sores in the mouth and on the palms and soles following a 3- to 7-day incubation period. This syndrome is rarely seen in adults. Case presentation A 35-year-old male Caucasian patient had a history of multiple episodes of acute pharyngitis, hypertension, hypercholesterolemia, and occasional abdominal pain. He presented with polyarthralgia in the knees and hands and odynophagia, followed by fever, oral mucosal aphthous lesions, and vesicles on the palms and soles. Three weeks after presentation, he was admitted to the emergency room with acute myocarditis. The in-hospital evaluation revealed positive serology for coxsackie A9 (1:160), positive anti-transglutaminase and anti-gliadin antibodies, normal immunoglobulins, and human immunodeficiency virus negativity. Conclusion We herein describe a case of HFMS that was associated with coxsackie A9 infection complicated by acute myocarditis. Although an association between celiac disease and HFMS has not been described, this patient’s immunologic disruption could have favored the development of infection and ultimately HFMS. PMID:24180226



Detection of bioagents using a shear horizontal surface acoustic wave biosensor  


A biosensor combining the sensitivity of surface acoustic waves (SAW) generated at a frequency of 325 MHz with the specificity provided by antibodies and other ligands for the detection of viral agents. In a preferred embodiment, a lithium tantalate based SAW transducer with silicon dioxide waveguide sensor platform featuring three test and one reference delay lines was used to adsorb antibodies directed against Coxsackie virus B4 or the negative-stranded category A bioagent Sin Nombre virus (SNV). Rapid detection of increasing concentrations of viral particles was linear over a range of order of magnitude for both viruses, and the sensor's selectivity for its target was not compromised by the presence of confounding Herpes Simplex virus type 1 The biosensor was able to delect SNV at doses lower than the load of virus typically found in a human patient suffering from hantavirus cardiopulmonary syndrome (HCPS).

Larson, Richard S; Hjelle, Brian; Hall, Pam R; Brown, David C; Bisoffi, Marco; Brozik, Susan M; Branch, Darren W; Edwards, Thayne L; Wheeler, David



[Onychomadesis associated with mouth, hand and foot disease].  


Onychomadesis is the spontaneous, complete shedding of the nail from its proximal side, without pain or inflammation, following nail matrix arrest. This disorder is uncommon in children and it can occur in fingernails, toenails or both. It may be secondary to systemic disorders, Kawasaki disease, bullous dermatoses, drugs, paronychia, stress and radiotherapy. Since 2000, Hand, Foot, and Mouth Disease (HFMD) has been described as a cause of onychomadesis, and has been associated with outbreaks of this condition in different regions of the world. HFMD is an infection characterized by vesicular and erosive stomatitis in combination with a vesicular eruption in palms and soles. It occurs in small children during summer and autumn months, and it is caused by coxsackie virus. We present a study that reflects the current situation of onychomadesis in Argentinian children and shows a strong association between this disorder and HFMD, suggesting that onychomadesis is a new manifestation of a previously known disease. PMID:24196774

Ferrari, Bruno; Taliercio, Vanina; Hornos, Lorena; Luna, Paula; Abad, María Eugenia; Larralde, Margarita



[Immunologic reactivity and status of the intestinal viral flora of humans in pressurized chambers].  


Immunity status and intestinal viral flora were studied in 5 experiments with long-term exposure of 26 male subjects in pressurized chamber. In two experiments with congenital microclimate and gaseous environment cellular and humoral indices of immunity did not deviate from the physiological norm. None of 14 subjects in these experiments was identified as a virus-carrier. In three experiments with hard environmental conditions (elevated concentrations of ammonia and carbon dioxide) viruses were isolated from 8 out of 12 subjects. Activation of viral flora mainly occurred on a background of impaired immunologic reactivity. In two experiments carriage of virus was asymptomatic; in the third experiment with the rise of ammonia up to 7 mg/m3 two subjects developed clinic signs that could have been engendered by enteric viruses, the Coxsackie B strain specifically. PMID:11816403

Antropova, E N; Rykova, M P; Meshkov, D O; Kazantseva, V A



The Molecular Interaction of CAR and JAML Recruits the Central Cell Signal Transducer PI3K  

SciTech Connect

Coxsackie and adenovirus receptor (CAR) is the primary cellular receptor for group B coxsackieviruses and most adenovirus serotypes and plays a crucial role in adenoviral gene therapy. Recent discovery of the interaction between junctional adhesion molecule-like protein (JAML) and CAR uncovered important functional roles in immunity, inflammation, and tissue homeostasis. Crystal structures of JAML ectodomain (2.2 angstroms) and its complex with CAR (2.8 angstroms) reveal an unusual immunoglobulin-domain assembly for JAML and a charged interface that confers high specificity. Biochemical and mutagenesis studies illustrate how CAR-mediated clustering of JAML recruits phosphoinositide 3-kinase (P13K) to a JAML intracellular sequence motif as delineated for the {alpha}{beta} T cell costimulatory receptor CD28. Thus, CAR and JAML are cell signaling receptors of the immune system with implications for asthma, cancer, and chronic nonhealing wounds.

Verdino, Petra; Witherden, Deborah A.; Havran, Wendy L.; Wilson, Ian A. (Scripps)



Antiviral activity of Folium isatidis derived extracts in vitro and in vivo.  


Folium isatidis is a native Chinese herbaceous plant widely used for medicinal purposes for thousands of years. However, few studies have focused on the leaves of Isatis indigotica. In this report, we isolated a series of four fractions (I-IV) from Folium isatidis and explored the antiviral activity of each tested extract. The extracts were active against a panel of RNA and DNA viruses in vitro, namely influenza A virus (IAV), coxsackie virus B3 (CVB3), respiratory syncytial virus (RSV), and adenovirus type 7 (Ad-7). Oral administration of 200 mg/kg/d of fraction III in mice exerted strong antiviral effects in viral replication, accompanied by prolonged survival rate, attenuated lung tissue damage as well as significant reductions in pulmonary virus titers and lung index. Our results provide the first biochemical evidence that Folium isatidis and its extracts could be used as potential antiviral agent in the postexposure prophylaxis for multiple viral infections. PMID:23895163

Deng, You-Ping; Liu, Yuan-Yuan; Liu, Zhao; Li, Jin; Zhao, Ling-Min; Xiao, Hong; Ding, Xiao-Hua; Yang, Zhan-Qiu



Identification of naturally processed T cell epitopes from glutamic acid decarboxylase presented in the context of HLA-DR alleles by T lymphocytes of recent onset IDDM patients.  

PubMed Central

Glutamic acid decarboxylase (GAD) has been defined as a major target antigen in insulin-dependent diabetes mellitus (IDDM). To identify the molecular ligands triggering a T cell response to GAD, a panel of human GAD65-specific T lymphocyte lines was generated from peripheral blood of three recent onset IDDM patients. All lines derived from a patient expressing the high-risk-conferring HLA-DR*0301/ *0401 haplotypes recognized a single epitope localized between amino acid positions 270 and 283 of GAD65, a stretch that is located in close proximity to the homology region shared with Coxsackie virus P2-C protein. All lines with this specificity were restricted to the DRA, B1*0401 product of the DR4 haplotype. Analysis of the GAD-specific T cell response in a second patient homozygous for DR4 haplotypes demonstrated that the same DRA, B1*0401 allele selected T cells specific for a different determinant. The T cell response profile in a third patient showed that DR*1501/ *1601-encoding haplotypes could present at least three different epitopes to GAD65-specific T lymphocytes. One of these epitopes was presented by a DR allele associated with the resistance-conferring DRB1*1501 haplotype. GAD-specific T cell lines could not be isolated from HLA class II-matched normal individuals. Our data reveal that (a) the T cell response to GAD65 is quite heterogenous in recent onset IDDM patients; (b) HLA-DR, not DQ, seems to be the principal restriction element used by T cells present at the onset of the disease; and (c) T cells responding to epitopes containing identical sequences to Coxsackie virus P2-C protein were not detected. PMID:9153283

Endl, J; Otto, H; Jung, G; Dreisbusch, B; Donie, F; Stahl, P; Elbracht, R; Schmitz, G; Meinl, E; Hummel, M; Ziegler, A G; Wank, R; Schendel, D J



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

PubMed Central

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

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



Rethinking Molecular Mimicry in Rheumatic Heart Disease and Autoimmune Myocarditis: Laminin, Collagen IV, CAR, and B1AR as Initial Targets of Disease  

PubMed Central

Rationale: Molecular mimicry theory (MMT) suggests that epitope mimicry between pathogens and human proteins can activate autoimmune disease. Group A streptococci (GAS) mimics human cardiac myosin in rheumatic heart disease (RHD) and coxsackie viruses (CX) mimic actin in autoimmune myocarditis (AM). But myosin and actin are immunologically inaccessible and unlikely initial targets. Extracellular cardiac proteins that mimic GAS and CX would be more likely. Objectives: To determine whether extracellular cardiac proteins such as coxsackie and adenovirus receptor (CAR), beta 1 adrenergic receptor (B1AR), CD55/DAF, laminin, and collagen IV mimic GAS, CX, and/or cardiac myosin or actin. Methods: BLAST 2.0 and LALIGN searches of the UniProt protein database were employed to identify potential molecular mimics. Quantitative enzyme-linked immunosorbent assay was used to measure antibody cross-reactivity. Measurements: Similarities were considered to be significant if a sequence contained at least 5 identical amino acids in 10. Antibodies were considered to be cross-reactive if the binding constant had a Kd less than 10-9 M. Main results: Group A streptococci mimics laminin, CAR, and myosin. CX mimics actin and collagen IV and B1AR. The similarity search results are mirrored by antibody cross-reactivities. Additionally, antibodies against laminin recognize antibodies against collagen IV; antibodies against actin recognize antibodies against myosin, and antibodies against GAS recognize antibodies against CX. Thus, there is both mimicry of extracellular proteins and antigenic complementarity between GAS-CX in RHD/AM. Conclusion: Rheumatic heart disease/AM may be due to combined infections of GAS with CX localized at cardiomyocytes that may produce a synergistic, hyperinflammatory response that cross-reacts with laminin, collagen IV, CAR, and/or B1AR. Epitope drift shifts the immune response to myosin and actin after cardiomyocytes become damaged. PMID:25191648

Root-Bernstein, Robert



Impact of the coxsackievirus and adenovirus receptor on the adenoma–carcinoma sequence of colon cancer  

PubMed Central

Background: Coxsackie and adenovirus receptor (CAR) has been suggested to function as a tumour suppressor. Its impact on the adenoma–carcinoma sequence of the colon, however, is unclear. Methods: Coxsackie and adenovirus receptor was analysed in non-cancerous and neoplastic colon samples using immunohistochemistry and quantitative RT–PCR. The function of CAR in colon cancer cell lines was determined following application of CAR siRNA or ectopic expression of a human full-length CAR cDNA. Results: Compared with healthy mucosa, increased CAR-mRNA expression was found in adenomas, whereas primary cancers and metastases displayed a marked decline. At the plasma membrane, CAR was present in normal mucosa samples (93%), adenomas, and metastases (100% ea.), whereas in colon cancers, it was found less frequently (49%, P<0.0001). Cytoplasmic CAR immunopositivity increased from normal mucosa (22%), to adenomas (73%, P=0.0006), primary cancers (83%, P<0.0001), and metastases (67%, P=0.0019). In cancer cell lines, CAR inhibition resulted in increased proliferation, whereas enforced ectopic CAR expression led to opposite results. Blocking the extracellular portion of CAR increased cell invasion in vitro. In mice, xenotransplants of colon cancer cells with enforced CAR expression formed significantly smaller tumours, whereas CAR inhibition increased the formation of liver metastases. Conclusion: We conclude that CAR facilitates complex effects during colon carcinogenesis, potentially mediated by its stage-dependent subcellular distribution; high CAR expression potentially prevents apoptosis in adenomas, loss of CAR at the plasma membrane promotes growth, and dissemination of primary cancers, and high membranous CAR presence may support the establishment of distant metastases. PMID:21468049

Stecker, K; Vieth, M; Koschel, A; Wiedenmann, B; Röcken, C; Anders, M



Screening for adenoviruses in haematological neoplasia: High prevalence in mantle cell lymphoma.  


Human adenoviruses possess oncogenic capacity which is well documented in mammalian animal models, but their possible implication in human malignancy has remained enigmatic. Following primary infection, adenoviruses can persist in a latent state in lymphocytes where the virus is apparently able to evade immune surveillance. In the present study, we have employed a broad-spectrum adenovirus polymerase chain reaction (PCR) assay to systematically screen more than 200 diagnostic specimens of different lymphoid malignancies including acute lymphocytic leukaemia (n=50), chronic lymphocytic leukaemia (n=50), various types of malignant lymphoma (n=100) and multiple myeloma (n=11) for the presence of adenoviral sequences. While most entities analysed revealed negative findings in virtually all specimens tested, adenoviral DNA was detected in 15/36 (42%) mantle cell lymphomas investigated. The most prevalent adenoviral species detected was C, and less commonly B. Adenovirus-positive findings in patients with mantle cell lymphoma were made at different sites including bone marrow (n=7), intestine (n=5), lymph nodes (n=2) and tonsillar tissue (n=1). The presence of adenoviral sequences identified by PCR was confirmed in individual cells by fluorescence in-situ hybridisation (FISH). The frequent observation of adenoviruses in mantle cell lymphoma is intriguings, and raises questions about their possible involvement in the pathogenesis of this lymphoid malignancy. PMID:24246703

Kosulin, Karin; Rauch, Margit; Ambros, Peter F; Pötschger, Ulrike; Chott, Andreas; Jäger, Ulrich; Drach, Johannes; Nader, Alexander; Lion, Thomas



Human adenovirus-host cell interactions: comparative study with members of subgroups B and C.  

PubMed Central

Host cell interactions of human adenovirus serotypes belonging to subgroups B (adenovirus type 3 [Ad3] and Ad7) and C (Ad2 and Ad5) were comparatively analyzed at three levels: (i) binding of virus particles with host cell receptors; (ii) cointernalization of macromolecules with adenovirions; and (iii) adenovirus-induced cytoskeletal alterations. The association constants with human cell receptors were found to be similar for Ad2 and Ad3 (8 x 10(9) to 9 x 10(9) M-1), and the number of receptor sites per cell ranged from 5,000 (Ad2) to 7,000 (Ad3). Affinity blottings, competition experiments, and immunofluorescence stainings suggested that the receptor sites for adenovirus were distinct for members of subgroups B and C. Adenovirions increased the permeability of cells to macromolecules. We showed that this global effect could be divided into two distinct events: (i) cointernalization of macromolecules and virions into endocytotic vesicles, a phenomenon that occurred in a serotype-independent way, and (ii) release of macromolecules into the cytoplasm upon adenovirus-induced lysis of endosomal membranes. The latter process was found to be type specific and to require unaltered and infectious virus particles of serotype 2 or 5. Perinuclear condensation of the vimentin filament network was observed at early stages of infection with Ad2 or Ad5 but not with Ad3, Ad7, and noninfectious particles of Ad2 or Ad5, obtained by heat inactivation of wild-type virions or with the H2 ts1 mutant. This phenomenon appeared to be a cytological marker for cytoplasmic transit of infectious virions within adenovirus-infected cells. It could be experimentally dissociated from vimentin proteolysis, which was found to be serotype dependent, occurring only with members of subgroup C, regardless of the infectivity of the input virus. Images PMID:2196380

Defer, C; Belin, M T; Caillet-Boudin, M L; Boulanger, P



1. Alternative splicing of viral receptors: A review of the diverse morphologies and physiologies of adenoviral receptors  

PubMed Central

Understanding the biology of cell surface proteins is important particularly when they are utilized as viral receptors for viral entry. By manipulating the expression of cell surface receptors that have been coopted by viruses, the susceptibility of an individual to virus-induced disease or, alternatively, the effectiveness of viral-based gene therapy can be modified. The most commonly studied vector for gene therapy is adenovirus. The majority of adenovirus types utilize the coxsackievirus and adenovirus receptor (CAR) as a primary receptor to enter cells. Species B adenovirus do not interact with CAR, but instead interact with the cell surface proteins desmoglein-2 (DSG-2) and cluster of differentiation 46 (CD46). These cell surface proteins exhibit varying degrees of alternative mRNA splicing, creating an estimated 20 distinct protein isoforms. It is likely that alternative splice forms have allowed these proteins to optimize their effectiveness in a plethora of niches, including roles as cell adhesion proteins and regulators of the innate immune system. Interestingly, there are soluble isoforms of these viral receptors, which lack the transmembrane domain. These soluble isoforms can potentially bind to the surface of a virus in the extracellular compartment, blocking the ability of the virus to bind to the host cell, reducing viral infectivity. Finally, the diversity of viral receptor isoforms appears to facilitate an assortment of interactions between viral receptor proteins and cytosolic proteins, leading to differential sorting in polarized cells. Using adenoviral receptors as a model system, the purpose of this review is to highlight the role that isoform-specific protein localization plays in the entry of pathogenic viruses from the apical surface of polarized epithelial cells.

Excoffon, Katherine J.D.A.; Bowers, Jonathan R.; Sharma, Priyanka



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

SciTech Connect

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

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



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


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

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



High sensitivity and label-free detection of Enterovirus 71 by nanogold modified electrochemical impedance spectroscopy  

NASA Astrophysics Data System (ADS)

Enterovirus 71 (EV71), which is the most fulminant and invasive species of enterovirus, can cause children neurologic complications and death within 2-3 days after fever and rash developed. Besides, EV71 has high sequence similarity with Coxsackie A 16 (CA16) that makes differential diagnosis difficult in clinic and laboratory. Since conventional viral diagnostic method cannot diagnose EV71 quickly and EV71 can transmit at low viral titer, the patients might delay in treatment. A quick, high sensitive, and high specific test for EV71 detection is pivotal. Electrochemical impedance spectroscopy (EIS) has been applied for detecting bio-molecules as biosensors recently. In this study, we try to build a detection platform for EV71 detection by nanogold modified EIS probe. The result shows that our probe can detect 3.6 VP1/50 ?l (one EV71 particle has 60 VP1) in 3 minutes. The test can also distinguish EV71 from CA16 and lysozyme. Diagnosis of enterovirus 71 by electrochemical impedance spectroscopy has the potential to apply in clinic.

Wang, Fang-Yu; Li, Hsing-Yuan; Tseng, Shing-Hua; Cheng, Tsai-Mu; Chu, Hsueh-Liang; Yang, Jyh-Yuan; Chang, Chia-Ching



Rapid and highly sensitive detection of Enterovirus 71 by using nanogold-enhanced electrochemical impedance spectroscopy.  


Enterovirus 71 (EV71) infection is an emerging infectious disease causing neurological complications and/or death within two to three days after the development of fever and rash. A low viral titre in clinical specimens makes the detection of EV71 difficult. Conventional approaches for detecting EV71 are time consuming, poorly sensitive, or complicated, and cannot be used effectively for clinical diagnosis. Furthermore, EV71 and Coxsackie virus A16 (CA16) may cross react in conventional assays. Therefore, a rapid, highly sensitive, specific, and user-friendly test is needed. We developed an EV71-specific nanogold-modified working electrode for electrochemical impedance spectroscopy in the detection of EV71. Our results show that EV71 can be distinguished from CA16, Herpes simplex virus, and lysozyme, with the modified nanogold electrode being able to detect EV71 in concentrations as low as 1 copy number/50 ?l reaction volume, and the duration between sample preparation and detection being 11 min. This detection platform may have the potential for use in point-of-care diagnostics. PMID:23787733

Li, Hsing-Yuan; Tseng, Shing-Hua; Cheng, Tsai-Mu; Chu, Hsueh-Liang; Lu, Yu-Ning; Wang, Fang-Yu; Tsai, Li-Yun; Shieh, Juo-Yu; Yang, Jyh-Yuan; Juan, Chien-Chang; Tu, Lung-Chen; Chang, Chia-Ching



Viral infection of engrafted human islets leads to diabetes.  


Type 1 diabetes (T1D) is characterized by the destruction of the insulin-producing ?-cells of pancreatic islets. Genetic and environmental factors both contribute to T1D development. Viral infection with enteroviruses is a suspected trigger for T1D but a causal role remains unproven and controversial. Studies in animals are problematic because of species-specific differences in host cell susceptibility and immune responses to candidate viral pathogens such as coxsackie B virus (CVB). In order to resolve the controversial role of viruses in human T1D, we developed a viral infection model in immunodeficient mice bearing human islet grafts. Hyperglycemia was induced in mice by specific ablation of native ?-cells. Human islets, which are naturally susceptible to CVB infection, were transplanted to restore normoglycemia. Transplanted mice were infected with CVB4 and monitored for hyperglycemia. Forty-seven percent of CVB4-infected mice developed hyperglycemia. Human islet grafts from infected mice contained viral RNA, expressed viral protein, and had reduced insulin compared to grafts from uninfected mice. Human-specific gene expression profiles in grafts from infected mice revealed the induction of multiple interferon stimulated genes. Thus, human islets can become severely dysfunctional with diminished insulin production following CVB infection of ?-cells, resulting in diabetes. PMID:25392246

Gallagher, Glen R; Brehm, Michael A; Finberg, Robert W; Barton, Bruce A; Shultz, Leonard D; Greiner, Dale L; Bortell, Rita; Wang, Jennifer P



Infectious muscle disease.  


Infectious muscle diseases have very different aetiologies. The viral myositides are proved by clinical and laboratory evidences in various etiologic settings (Influenza A and B, Coxsackie and HIV). The bacterial myositis was considered in the near past a tropical disease, but in our days with migration of people from South to North and the endemia of AIDS it became a problem of the "civilized" world. On the other hand, tuberculous endemia in Central-Eastern Europe, including Romania, results in quite high incidence of osteoarticular tuberculosis. In this section the authors take into consideration some clinical entities, such as psoas abscess, postanginal sepsis, beta-haemolytic streptococcus infection and that caused by Koch bacillus. Other rare musculoskeletal infections such as gas gangrene and non-clostridial anaerobic myonecrosis are also reviewed. Immune depression caused by underlying diseases, therapies, alcoholism or old age is often encountered. The parasitic aetiologies include infestations with Trichinella spiralis, Cysticercus cellulosae, Toxoplasma and Amoeba. The contribution of imagistic methods to diagnosis is emphasised. Ultrasonography associated with CT imaging are usually used, while MRI should be reserved for cases in which axial skeleton is involved. The management is based on appropriate antibiotic therapy and surgery. PMID:17236294

Parasca, I; Damian, Laura; Albu, Adriana



EGFR-Targeted Adenovirus Dendrimer Coating for Improved Systemic Delivery of the Theranostic NIS Gene  

PubMed Central

We recently demonstrated tumor-selective iodide uptake and therapeutic efficacy of combined radiovirotherapy after systemic delivery of the theranostic sodium iodide symporter (NIS) gene using a dendrimer-coated adenovirus. To further improve shielding and targeting we physically coated replication-selective adenoviruses carrying the hNIS gene with a conjugate consisting of cationic poly(amidoamine) (PAMAM) dendrimer linked to the peptidic, epidermal growth factor receptor (EGFR)-specific ligand GE11. In vitro experiments demonstrated coxsackie-adenovirus receptor-independent but EGFR-specific transduction efficiency. Systemic injection of the uncoated adenovirus in a liver cancer xenograft mouse model led to high levels of NIS expression in the liver due to hepatic sequestration, which were significantly reduced after coating as demonstrated by 123I-scintigraphy. Reduction of adenovirus liver pooling resulted in decreased hepatotoxicity and increased transduction efficiency in peripheral xenograft tumors. 124I-PET-imaging confirmed EGFR-specificity by significantly lower tumoral radioiodine accumulation after pretreatment with the EGFR-specific antibody cetuximab. A significantly enhanced oncolytic effect was observed following systemic application of dendrimer-coated adenovirus that was further increased by additional treatment with a therapeutic dose of 131I. These results demonstrate restricted virus tropism and tumor-selective retargeting after systemic application of coated, EGFR-targeted adenoviruses therefore representing a promising strategy for improved systemic adenoviral NIS gene therapy. PMID:24193032

Grünwald, Geoffrey K; Vetter, Alexandra; Klutz, Kathrin; Willhauck, Michael J; Schwenk, Nathalie; Senekowitsch-Schmidtke, Reingard; Schwaiger, Markus; Zach, Christian; Wagner, Ernst; Göke, Burkhard; Holm, Per S; Ogris, Manfred; Spitzweg, Christine



An adenoviral vector for probing promoter activity in primary immune cells  

PubMed Central

Functional analysis of the DNA regulatory regions that control gene expression has largely been performed through transient transfection of promoter–reporter constructs into transformed cells. However, transformed cells are often poor models of primary cells. To directly analyze DNA regulatory regions in primary cells, we generated a novel adenoviral luciferase reporter vector, pShuttle-luciferase-GFP (pSLUG) that contains a promoterless luciferase cassette (with an upstream cloning site) for probing promoter activity, and a GFP expression cassette that allows for the identification of transduced cells. Recombinant adenoviruses generated from this vector can transduce a wide range of primary immune cells with high efficiency, including human macrophages, dendritic cells and T cells; and mouse T cells transgenic for the coxsackie and adenoviral receptor (CAR). In primary T cells, we show inducible nuclear factor of activated T cells (NF-AT) activity using a recombinant pSLUG adenovirus containing a consensus NF-AT promoter. We further show inducible IL-12/23 p40 promoter activity in primary macrophages and dendritic cells using a recombinant pSLUG adenovirus containing the proximal human IL-12/23 p40 promoter. The pSLUG system promises to be a powerful tool for the analysis of DNA regulatory regions in diverse types of primary immune cells. PMID:16563424

Tripathi, Pulak; Madan, Rajat; Chougnet, Claire; Divanovic, Senad; Ma, Xiaojing; Wahl, Larry M.; Gajewski, Thomas; Karp, Christopher L.; Hildeman, David A.



Enhanced Prostate Cancer Gene Transfer and Therapy Using a Novel Serotype Chimera Cancer Terminator Virus (Ad.5/3-CTV)  

PubMed Central

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




Recurrent pericarditis: infectious or autoimmune?  


The etiology and pathogenesis of idiopathic recurrent acute pericarditis (IRAP) remain controversial standing like a bridge that crosses infectious, autoimmune and autoinflammatory pathways. Anything may cause acute pericarditis; Echo-virus, and Coxsackie are the most frequently involved viruses, Mycobacterium tuberculosis and Coxiella burnetii the most common bacteria, but in 85% of cases it remains "idiopathic". Recurrences occur in up to 20-50% of patients. An immuno-mediated pathogenesis is suggested by the presence of pro-inflammatory cytokines in pericardial fluid, the presence of antinuclear autoantibodies (ANA) in sera of the patients, the occurrence of new autoimmune diagnoses and the good response to anti-inflammatory or immunosuppressive therapy. Nonsteroidal anti-inflammatory drugs (NSAIDs) must be used at recommended dosages, till the resolution of symptoms and normalization of C-reactive protein and erythrocyte sedimentation rate. Corticosteroids should be used rarely, at low doses, with an extremely low tapering and with osteoporosis prevention. Colchicine leads to a clinically important and statistically significant benefit, reducing recurrences by 50%. The long term outcome of IRAP is good, without evidence of constriction even after a very long follow-up. PMID:18708165

Brucato, Antonio; Maestroni, Silvia; Cumetti, Davide; Thiella, Giuseppe; Alari, Gabriella; Brambilla, Giovanni; Imazio, Massimo; Doria, Andrea; Palmieri, Giancarlo; Adler, Yehuda



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


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

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



Rapid and highly sensitive detection of Enterovirus 71 by using nanogold-enhanced electrochemical impedance spectroscopy  

NASA Astrophysics Data System (ADS)

Enterovirus 71 (EV71) infection is an emerging infectious disease causing neurological complications and/or death within two to three days after the development of fever and rash. A low viral titre in clinical specimens makes the detection of EV71 difficult. Conventional approaches for detecting EV71 are time consuming, poorly sensitive, or complicated, and cannot be used effectively for clinical diagnosis. Furthermore, EV71 and Coxsackie virus A16 (CA16) may cross react in conventional assays. Therefore, a rapid, highly sensitive, specific, and user-friendly test is needed. We developed an EV71-specific nanogold-modified working electrode for electrochemical impedance spectroscopy in the detection of EV71. Our results show that EV71 can be distinguished from CA16, Herpes simplex virus, and lysozyme, with the modified nanogold electrode being able to detect EV71 in concentrations as low as 1 copy number/50 ?l reaction volume, and the duration between sample preparation and detection being 11 min. This detection platform may have the potential for use in point-of-care diagnostics.

Li, Hsing-Yuan; Tseng, Shing-Hua; Cheng, Tsai-Mu; Chu, Hsueh-Liang; Lu, Yu-Ning; Wang, Fang-Yu; Tsai, Li-Yun; Shieh, Juo-Yu; Yang, Jyh-Yuan; Juan, Chien-Chang; Tu, Lung-Chen; Chang, Chia-Ching





Rabbit sera were found to possess neutralizing activity (normal antibody) to polioviruses and Coxsackie B viruses. This normal antibody showed high specificity in cross-neutralization and absorption tests. It was associated with heat-stable, mercaptan-sensitive, 19S gamma(1)-beta-macroglobulins, which formed weak complexes with the viral antigen. In rare instances, sera with normal macroglobulin antibody, also contained very low activity which was due to 7S gamma(2)-globulins. The neutralization of poliovirus by normal 19S gamma(1)-beta-antibody appeared to follow first order kinetics, and the thermodynamic parameters of this reaction were the same as those of serological reactions employing immune antibody. The electrophoretic mobility, sedimentation properties, sensitivity to mercaptan, thermostability, and avidity of normal and early (up to day 3) immune antibodies to poliovirus were similar, but differed in several respects from those of late immune antibodies. Thus, the available evidence suggested, that earlier reported differences between normal and immune antibodies reflected differences between antibodies of diverse physicochemical properties rather than between normal and immune antibodies per se. It is proposed that the normal macroglobulin antibody is associated with an immunological response to repeated stimulation with minute amounts of antigen. PMID:14151096




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


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

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



Retargeted adenoviral cancer gene therapy for tumour cells overexpressing epidermal growth factor receptor or urokinase-type plasminogen activator receptor  

PubMed Central

We have assessed the ability of bispecific fusion proteins to improve adenovirus-mediated transfer of therapeutic and marker transgenes. We constructed an expression vector that can be easily modified to synthesize a variety of fusion proteins for retargeting adenoviral gene therapy vectors to cell surface markers, which are differentially expressed between normal and cancer cells. Adenoviral transduction can be improved in a number of tumour cell lines which overexpress EGFR (epidermal growth factor receptor) or uPAR (urokinase-type plasminogen activator receptor), but which have only low levels of endogenous hCAR (human coxsackie B and adenovirus receptor) expression. Up to 40-fold improvement in ?-galactosidase transgene expression was seen using an EGFR retargeting protein, and up to 16-fold using a second fusion protein targeting uPAR. In vitro, our uPAR retargeting fusion protein improved the sensitivity to adenoviral herpes simplex virus thymidine kinase/ganciclovir by an order of magnitude, whereas in vivo, our EGFR retargeting protein is able to significantly delay tumour growth in rodent animal models in a dose-dependent manner. The ‘cassette’ design of our fusion protein constructs offers a flexible method for the straightforward synthesis of multiple adenoviral retargeting proteins, directed against a variety of tumour-associated antigens, for use in clinical trials. PMID:20410926

Harvey, TJ; Burdon, D; Steele, L; Ingram, N; Hall, GD; Selby, PJ; Vile, RG; Cooper, PA; Shnyder, SD; Chester, JD



Oncolytic adenovirus modified with somatostatin motifs for selective infection of neuroendocrine tumor cells.  


We have previously described the oncolytic adenovirus, Ad(CgA-E1A-miR122), herein denoted Ad5(CgA-E1A-miR122) that selectively replicates in and kills neuroendocrine cells, including freshly isolated midgut carcinoid cells from liver metastases. Ad5(CgA-E1A-miR122) is based on human adenovirus serotype 5 (Ad5) and infects target cells by binding to the coxsackie-adenovirus receptor (CAR) and integrins on the cell surface. Some neuroendocrine tumor (NET) and neuroblastoma cells express low levels of CAR and are therefore poorly transduced by Ad5. However, they often express high levels of somatostatin receptors (SSTRs). Therefore, we introduced cyclic peptides, which contain four amino acids (FWKT) and mimic the binding site for SSTRs in the virus fiber knob. We show that FWKT-modified Ad5 binds to SSTR? on NET cells and transduces midgut carcinoid cells from liver metastases about 3-4 times better than non-modified Ad5. Moreover, FWKT-modified Ad5 overcomes neutralization in an ex vivo human blood loop model to greater extent than Ad5, indicating that fiber knob modification may prolong the systemic circulation time. We conclude that modification of adenovirus with the FWKT motif may be beneficial for NET therapy. PMID:21490682

Leja, J; Yu, D; Nilsson, B; Gedda, L; Zieba, A; Hakkarainen, T; Ĺkerström, G; Öberg, K; Giandomenico, V; Essand, M



Derivation of a myeloid cell-binding adenovirus for gene therapy of inflammation.  


The gene therapy field is currently limited by the lack of vehicles that permit efficient gene delivery to specific cell or tissue subsets. Native viral vector tropisms offer a powerful platform for transgene delivery but remain nonspecific, requiring elevated viral doses to achieve efficacy. In order to improve upon these strategies, our group has focused on genetically engineering targeting domains into viral capsid proteins, particularly those based on adenovirus serotype 5 (Ad5). Our primary strategy is based on deletion of the fiber knob domain, to eliminate broad tissue specificity through the human coxsackie-and-adenovirus receptor (hCAR), with seamless incorporation of ligands to re-direct Ad tropism to cell types that express the cognate receptors. Previously, our group and others have demonstrated successful implementation of this strategy in order to specifically target Ad to a number of surface molecules expressed on immortalized cell lines. Here, we utilized phage biopanning to identify a myeloid cell-binding peptide (MBP), with the sequence WTLDRGY, and demonstrated that MBP can be successfully incorporated into a knob-deleted Ad5. The resulting virus, Ad.MBP, results in specific binding to primary myeloid cell types, as well as significantly higher transduction of these target populations ex vivo, compared to unmodified Ad5. These data are the first step in demonstrating Ad targeting to cell types associated with inflammatory disease. PMID:22624065

Alberti, Michael O; Roth, Justin C; Ismail, Mourad; Tsuruta, Yuko; Abraham, Edward; Pereboeva, Larisa; Gerson, Stanton L; Curiel, David T



A novel immunocompetent murine model for replicating oncolytic adenoviral therapy  

PubMed Central

Oncolytic adenoviruses are under investigation as a promising novel strategy for cancer immunotherapeutics. Unfortunately, there is no immunocompetent mouse cancer model to test oncolytic adenovirus because murine cancer cells are generally unable to produce infectious viral progeny from human adenoviruses. We find that the murine K-ras-induced lung adenocarcinoma cell line ADS-12 supports adenoviral infection and generates infectious viral progeny. ADS-12 cells express the coxsackie and adenovirus receptor and infected ADS-12 cells express the viral protein E1A. We find that our previously described oncolytic virus, adenovirus TAV-255 (AdTAV-255), kills ADS-12 cells in a dose- and time-dependent manner. We investigated ADS-12 cells as an in-vivo model system for replicating oncolytic adenoviruses. Subcutaneous injection of ADS-12 cells into immunocompetent 129 mice led to tumor formation in all injected mice. Intratumoral injection of AdTAV-255 in established tumors causes a significant reduction in tumor growth. This model system represents the first fully immunocompetent mouse model for cancer treatment with replicating oncolytic adenoviruses, and therefore will be useful to study the therapeutic effect of oncolytic adenoviruses in general and particularly immunostimulatory viruses designed to evoke an antitumor immune response. PMID:25525035

Zhang, L; Hedjran, F; Larson, C; Perez, G L; Reid, T



Interaction with Decay-Accelerating Factor Facilitates Coxsackievirus B Infection of Polarized Epithelial Cells  

PubMed Central

All coxsackie B (CB) viruses can initiate infection by attaching to the coxsackievirus and adenovirus receptor (CAR). Although some CB isolates also bind to decay-accelerating factor (DAF), the role of DAF interaction during infection remains uncertain. We recently observed that CAR in polarized epithelial cells is concentrated at tight junctions, where it is relatively inaccessible to virus. In the experiments reported here we found that, unlike CAR, DAF was present on the apical surface of polarized cells and that DAF-binding isolates of CB3 and CB5 infected polarized epithelial cells more efficiently than did isolates incapable of attaching to DAF. Virus attachment and subsequent infection of polarized cells by DAF-binding isolates were prevented in the presence of anti-DAF antibody. Serial passage on polarized cell monolayers selected for DAF-binding virus variants. Taken together, these results indicate that interaction with DAF on the apical surface of polarized epithelial cells facilitates infection by a subset of CB virus isolates. The results suggest a possible role for DAF in infection of epithelial cells at mucosal surfaces. PMID:12186929

Shieh, Joseph T. C.; Bergelson, Jeffrey M.



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


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

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



Elicitation of T-cell responses by structural and non-structural proteins of coxsackievirus B4.  


Coxsackievirus B4 (CV-B4) belongs to the genus Enterovirus within the family Picornaviridae. To investigate target proteins recognized by T-cells in human enterovirus B infections, virus-encoded structural [VP0 (VP4 and VP2), VP1, VP3] and non-structural (2A, 2B, 2C, 3C and 3D) proteins were expressed and purified in Escherichia coli. Peripheral blood of 19 healthy adult donors was used to create enterovirus-specific T-cell lines by repeated stimulation with CV-B4 cell lysate antigen. T-cell lines responded in individual patterns, and responses to all purified proteins were observed. The most often recognized enteroviral protein was VP0, which is the fusion between the most conserved structural proteins, VP4 and VP2. T-cell responses to VP0 were detected in 15 of the 19 (79?%) donor lines. Non-structural 2C protein was recognized in 11 of the 19 (58?%) lines, and 11 of the 19 (58?%) lines also had a response to 3D protein. Furthermore, responses to other non-structural proteins (2A, 2B and 3C) were also detected. T-cell responses did not correlate clearly to the individual HLA-DR-DQ phenotype or the history of past coxsackie B virus infections of the donors. PMID:25381056

Bengs, Suvi; Marttila, Jane; Susi, Petri; Ilonen, Jorma



Alkaloids from the root of Isatis indigotica.  


Seventeen new alkaloids (1-17) and 14 known analogues have been isolated from an aqueous extract of the root of Isatis indigotica. The structures and absolute configurations of these compounds were determined by extensive spectroscopic data analysis, including 2D NMR, single-crystal X-ray crystallography using anomalous scattering of Cu K? radiation, and electronic circular dichroism spectra calculations based on the quantum-mechanical time-dependent density functional theory. Compounds 1, 2, and 3 are the first examples of natural products with unique linkages between a molecule of 2-(4-methoxy-1H-indol-3-yl)acetonitrile and 2-(1H-indol-3-yl)acetonitrile, 2-(4-methoxy-1H-indol-3-yl)acetonitrile, and 4-hydroxyphenylethane, respectively. Compounds (-)-4 and (+)-4 represent the first natural products with the pyrrolo[2,3-b]indolo[5,5a,6-b,a]quinazoline skeleton. Some structural assignments for the new alkaloids suggest that the assignments made for certain previously reported alkaloids require revision. Compounds 1-3 and arvelexin (18) show antiviral activity against the influenza virus A/Hanfang/359/95 (H3N2), with IC(50) values of 3.70-12.35 ?M, and 17 inhibits Coxsackie virus B3 replication with an IC(50) of 6.87 ?M. PMID:22694318

Chen, Minghua; Gan, Lishe; Lin, Sheng; Wang, Xiaoliang; Li, Li; Li, Yuhuan; Zhu, Chenggen; Wang, Yanan; Jiang, Bingya; Jiang, Jiandong; Yang, Yongchun; Shi, Jiangong



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

PubMed Central

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

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



Impairment of Myocardial Mitochondria in Viral Myocardial Disease and Its Reflective Window in Peripheral Cells  

PubMed Central

Background Viral myocardial disease (VMD) is a common disease inducing heart failure. It has not been clear the roles of mitochondrial damage in the pathological changes of cardiomyocytes in VMD. Methods Myocardial tissues and lymphocytes were collected from 83 VMD patients. Control groups included 12 cases of healthy accidental death with myocardial autopsy and 23 healthy blood donors. The mouse model of viral myocarditis (VMC) was established by Coxsackie virus B3 infection and myocardial tissues and skeletal muscle were collected. Mitochondrial DNA (mtDNA) deletion rate was quantitatively determined using polymerase chain reaction. Results There was significantly difference of myocardial mitochondrial DNA deletion rate between VMD or VMC group and control group (P<0.05). Moreover, the loss of mitochondrial membrane phospholipids was significantly different between VMD or VMC group and control group. In VMC mice, there were negative correlations between myocardial mtDNA3867 deletion rate and left ventricular peak systolic pressure (LVPSP) (r?=??0.66, P<0.05), and between myocardial mtDNA3867 deletion rate and +dp/dtmax (r?=??0.79, P<0.05), while there was positive correlation between myocardial mtDNA3867 deletion rate and ?dp/dtmax (r?=?0.80, P<0.05). Conclusion Mitochondrial damage is an important pathophysiological mechanism leading to myocardial injury and cardiac dysfunction. The mitochondrial damage in the skeletal muscle and lymphocytes reflect a “window” of myocardial mitochondrial damage. PMID:25551390

Wei, Jin; Gao, Deng-Feng; Wang, Hao; Yan, Rui; Liu, Zhi-Quan; Yuan, Zu-Yi; Liu, Jian; Chen, Ming-Xia



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

PubMed Central

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

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



Recombination in Circulating Enteroviruses  

PubMed Central

Recombination is a well-known phenomenon for enteroviruses. However, the actual extent of recombination in circulating nonpoliovirus enteroviruses is not known. We have analyzed the phylogenetic relationships in four genome regions, VP1, 2A, 3D, and the 5? nontranslated region (NTR), of 40 enterovirus B strains (coxsackie B viruses and echoviruses) representing 11 serotypes and isolated in 1981 to 2002 in the former Soviet Union states. In the VP1 region, strains of the same serotype expectedly grouped with their prototype strain. However, as early as the 2A region, phylogenetic grouping differed significantly from that in the VP1 region and indicated recombination within the 2A region. Moreover, in the 5? NTR and 3D region, only 1 strain of 40 grouped with its prototype strain. Instead, we observed a major group in both the 5? NTR and the 3D region that united most (in the 5? NTR) or all (in the 3D region) of the strains studied, regardless of the serotype. Subdivision within that major group in the 3D region correlated with the time of virus isolation but not with the serotype. Therefore, we conclude that a majority, if not all, circulating enterovirus B strains are recombinants relative to the prototype strains, isolated mostly in the 1950s. Moreover, the ubiquitous recombination has allowed different regions of the enterovirus genome to evolve independently. Thus, a novel model of enterovirus genetics is proposed: the enterovirus genome is a stable symbiosis of genes, and enterovirus species consist of a finite set of capsid genes responsible for different serotypes and a continuum of nonstructural protein genes that seem to evolve in a relatively independent manner. PMID:12970427

Lukashev, Alexander N.; Lashkevich, Vasilii A.; Ivanova, Olga E.; Koroleva, Galina A.; Hinkkanen, Ari E.; Ilonen, Jorma



Prevalence of nonpolio enteroviruses in the sewage of Guangzhou city, China, from 2009 to 2012.  


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

Zheng, Huanying; 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



Dual tumor targeting with pH-sensitive and bioreducible polymer-complexed oncolytic adenovirus.  


Oncolytic adenoviruses (Ads) have shown great promise in cancer gene therapy but their efficacy has been compromised by potent immunological, biochemical, and specific tumor-targeting limitations. To take full advantage of the innate cancer-specific killing potency of oncolytic Ads but also exploit the subtleties of the tumor microenvironment, we have generated a pH-sensitive and bio-reducible polymer (PPCBA)-coated oncolytic Ad. Ad-PPCBA complexes showed higher cellular uptake at pH 6.0 than pH 7.4 in both high and low coxsackie and adenovirus receptor-(CAR)-expressing cells, thereby demonstrating Ad-PPCBA's ability to target the low pH hypoxic tumor microenvironment and overcome CAR dependence for target cell uptake. Endocytic mechanism studies indicated that Ad-PPCBA internalization is mediated by macropinocytosis instead of the CAR-dependent endocytic pathway that internalizes naked Ad. VEGF-specific shRNA-expressing oncolytic Ad complexed with PPCBA (RdB/shVEGF-PPCBA) elicited much more potent suppression of U87 human brain cancer cell VEGF gene expression in vitro, and human breast cancer MCF7 cell/Matrigel plug vascularization in a mouse model, when cancer cells had been previously infected at pH 6.0 versus pH 7.4. Moreover, intratumorally and intravenously injected RdB/shVEGF-PPCBA nanocomplexes elicited significantly higher therapeutic efficacy than naked virus in U87-tumor mouse xenograft models, reducing IL-6, ALT, and AST serum levels. These data demonstrated PPCBA's biocompatibility and capability to shield the Ad surface to prevent innate immune response against Ad after both intratumoral and systemic administration. Taken together, these results demonstrate that smart, tumor-specific, oncolytic Ad-PPCBA complexes can be exploited to treat both primary and metastatic tumors. PMID:25522965

Moon, Chang Yoon; Choi, Joung-Woo; Kasala, Dayananda; Jung, Soo-Jung; Kim, Sung Wan; Yun, Chae-Ok



Human cytomegalovirus in the pancreas of patients with type 2 diabetes: is there a relation to clinical features, mRNA and protein expression of insulin, somatostatin, and MHC class II?  


Human cytomegalovirus (HCMV) was recently demonstrated in the pancreas of about half the patients with type 2 diabetes mellitus in the absence of mumps, rubella or Coxsackie B virus. The present study addresses the question as to whether type 2 diabetes with an HCMV-positive pancreas differs from those with HCMV-negative pancreases with respect to age, sex, treatment, duration of disease, volume densities of B-cells and D-cells, mRNA levels of insulin and somatostatin, islet amyloid peptide deposits and major histocompatibility complex (MHC) class I and class II gene transcription, and protein expression. HCMV-positive type 2 diabetic patients showed a tendency towards a shorter duration of disease and significantly increased levels of MHC class II on RNA. In addition, expression of MHC class II product (HLA-DR) was identified in duct epithelial cells and/or islet cells in 9 diabetic pancreases and in 2 non-diabetic glands. No MHC class I expression could be detected. No other clinical differences between HCMV-positive and HCMV-negative glands were found. All 10 HCMV-positive diabetics showed a strong expression of MHC class II mRNA in the pancreas. By immunocytochemistry, 4 of 10 demonstrated expression on the islets; three of ten also expressed MHC DR beta on ductal cells. This finding might be related to the viral infection, as only 2 of the 9 HCMV-negative patients were HLA-DR beta positive and none of the non-diabetic controls showed increased levels of MHC class II mRNA. These data suggest that HCMV infection in the pancreas is associated with type 2 diabetes. However, no conclusions as to a role of this virus in the aetiopathology of type 2 diabetes can be drawn at present. PMID:1360719

Löhr, M; Bergstrome, B; Maekawa, R; Oldstone, M B; Klöppel, G



Prevalence of human enteroviruses among apparently healthy nursery school children in Accra  

PubMed Central

Introduction Human enteroviruses are common in children causing asymptomatic infections ranging from mild to severe illnesses. In Ghana, information on the prevalence of non-polio enterovirus causing acute flaccid paralysis is available but data on surveillance of these viruses in school children is scanty. Here, the prevalence of human enteroviruses among apparently healthy children in selected school in Accra was studied. Methods Stool samples from 273 apparently healthy children less than eight years of age in 9 selected nursery schools were collected between December 2010 and March 2011and processed for human enteroviruses on L20B, RD and Hep-2 cell lines. Positive Isolates were characterized by microneutralisation assay with antisera pools from RIVM, the Netherlands according to standard methods recommended by WHO. Results Of the 273 samples processed, 66 (24.2%) non-polio enteroviruses were isolated. More growth was seen on Hep-2C (46%) only than RD (18%) only and on both cell lines (34%). No growth was seen on L20B even after blind passage. Excretion of non-polio enteroviruses was found in all the schools with majority in BD school. Serotyping of the isolates yielded predominantly Coxsackie B viruses followed by echoviruses 13 and 7. More than half of the isolates could not be typed by the antisera pools. Conclusion The study detected 13 different serotypes of non-polio enteroviruses in circulation but no poliovirus was found. BD school was found to have the highest prevalence of NPEV. Complete identification through molecular methods is essential to establish the full range of NPEVs in circulation in these schools. PMID:25400833

Attoh, Juliana; Obodai, Evangeline; Adiku, Theophilus; Odoom, John Kofi



Nonhomologous Recombination between Defective Poliovirus and Coxsackievirus Genomes Suggests a New Model of Genetic Plasticity for Picornaviruses  

PubMed Central

ABSTRACT Most of the circulating vaccine-derived polioviruses (cVDPVs) implicated in poliomyelitis outbreaks in Madagascar have been shown to be recombinants between the type 2 poliovirus (PV) strain of the oral polio vaccine (Sabin 2) and another species C human enterovirus (HEV-C), such as type 17 coxsackie A virus (CA17) in particular. We studied intertypic genetic exchanges between PV and non-PV HEV-C by developing a recombination model, making it possible to rescue defective type 2 PV RNA genomes with a short deletion at the 3? end by the cotransfection of cells with defective or infectious CA17 RNAs. We isolated over 200 different PV/CA17 recombinants, using murine cells expressing the human PV receptor (PVR) and selecting viruses with PV capsids. We found some homologous (H) recombinants and, mostly, nonhomologous (NH) recombinants presenting duplications of parental sequences preferentially located in the regions encoding proteins 2A, 2B, and 3A. Short duplications appeared to be stable, whereas longer duplications were excised during passaging in cultured cells or after multiplication in PVR-transgenic mice, generating H recombinants with diverse sites of recombination. This suggests that NH recombination events may be a transient, intermediate step in the generation and selection of the fittest H recombinants. In addition to the classical copy-choice mechanism of recombination thought to generate mostly H recombinants, there may also be a modular mechanism of recombination, involving NH recombinant precursors, shaping the genomes of recombinant enteroviruses and other picornaviruses. PMID:25096874

Holmblat, Barbara; Jégouic, Sophie; Muslin, Claire; Blondel, Bruno; Joffret, Marie-Line



[Chemical consitituents from root of Isatis indigotica].  


Thirty-three compounds were isolated from the root decoction of Isatis indigotica by using a combination of various chromatographic techniques including silica gel, macroporous adsorbent resin, Sephadex LH-20, and reversed-phase HPLC. Their structures were elucidated by spectroscopic data as (+)-dehydrovomifoliol (1), (S)-(+)-abscisic acid (2), vomifoliol (3), cyclo (L-Phe-L-Leu) (4), cyclo(L-Phe-L-Tyr) (5), cyclo(L-Tyr-L-Leu) (6), cyclo(L-Pro-L-Tyr) (7), evofolin B (8), (+)-syringaresinol (9), (-)-(7R,7'R,8S,8'S)-4,4'-dihydroxy-3-methoxy-7,9';7',9-diepoxy-lignan (10), (-)-medioresinol (11), (+) -(7R,7'R,8S,8'S) -neo-olivil (12), (-) -5-methoxyisolariciresinol (13), 1,3-dihydro-2H-indol-2-one (14), isalexin (15), dihydroneoascorbigen (16), indican (17), (-) -(S) -cyanomethyl-3-hydroxyoxindole (18), isoformononetein (19), calycosin (20), stigamast-5-ene-3beta-ol-7-one (21), acetovanillone (22), 3, 5-dimethoxy-4-hydroxyacetophenone (23), dihydroconiferyl alcohol (24), dihyroferulic acid (25), 3-hydroxy-1-(4-hydroxyphenyl) propan-1-one (26), beta-hydroxypropiovanillone (27), 4-aminobenzoic acid (28), 3-(4-hydroxyphenyl) propan-1-ol (29), 4-(2-hydroxyethyl) phenol (30), 2-methoxy-4-vinylphenol (31), pyrocatechol (32), and 4-pentenamide (33). These compounds were isolated from the root of I. indigotica for the first time. In preliminary in vitro assays, compound 19 showed activity against the influenza virus A/Hanfang/359/95 (H3N2), the herpes simplex virus 1 (HSV-1), and Coxsackie virus B3 (Cox-B3), with IC50 values of 2.06, 6.84, and 8.70 micromol x L(-1), respectively, but other compounds were in-active at a concentration of 1.0 x 10 x (-5) mol x L(-1). PMID:23944031

Wang, Xiao-Liang; Chen, Ming-Hua; Wang, Fang; Bu, Peng-Bin; Lin, Sheng; Zhu, Cheng-Gen; Li, Yu-Huan; Jiang, Jian-Dong; Shi, Jian-Gong



Mutation in fiber of adenovirus serotype 5 gene therapy vector decreases liver tropism  

PubMed Central

Recombinant adenovirus (Ad) vectors are widely used for both in vitro and in vivo gene transfer. However, intravenous administration of Ad vectors results mainly in hepatocyte transduction and subsequent hepatotoxicity. Coxsackie-adenovirus receptor (CAR) and ?v? integrins, which are functional receptors for the fiber and penton proteins, respectively, are the tropism determinants of Ad type 5 (Ad5). We previously developed a system for rapid construction of fiber-modified Ad5 vectors. We also constructed a fiber-modified Ad5 containing an Arg-Gly-Asp (RGD) motif in the HI-loop and showed that it could enhance anti-tumor effects in vitro and in vivo. Here, we constructed a novel Ad5 vector containing two amino acid mutations in the AB loop of the fiber-modified Ad5 fiber knob and showed that it could significantly reduce liver tropism and increase gene transfer in low-CAR or CAR-deficient cancer cells following intravascular delivery. However, anti-tumor effects of the fiber-mutated Ad5 expressing HSV-TK under control of the hTERT promoter was not found when compared with an unmodified Ad5 vector in cancer lines expressing different levels of CAR, likely due to the activity of the hTERT promoter being lower than that of the CMV promoter. Nevertheless, this study describes an enhanced Ad5 vector for intravascular gene delivery, and further modifications such as changes in the promoter may facilitate the development of this vector for cancer treatment.

Wang, Zhen; Wang, Baoming; Lou, Junfang; Yan, Jingyi; Gao, Lei; Geng, Ranshen; Yu, Bin



Safety Profiles and Antitumor Efficacy of Oncolytic Adenovirus Coated with Bioreducible Polymer in the Treatment of a CAR Negative Tumor Model.  


Adenovirus (Ad) vectors show promise as cancer gene therapy delivery vehicles, but immunogenic safety concerns and coxsackie and adenovirus receptor (CAR)-dependency have limited their use. Alternately, biocompatible and bioreducible nonviral vectors, including arginine-grafted cationic polymers, have been shown to deliver nucleic acids through a cell penetration peptide (CPP) and protein transduction domain (PTD) effect. We utilized the advantages of both viral and nonviral vectors to develop a hybrid gene delivery vehicle by coating Ad with mPEG-PEI-g-Arg-S-S-Arg-g-PEI-mPEG (Ad/PPSA). Characterization of Ad/PPSA particle size and zeta potential showed an overall size and cationic charge increase in a polymer concentration-dependent manner. Ad/PPSA also showed a marked transduction efficiency increase in both CAR-negative and -positive cells compared to naked Ad. Competition assays demonstrated that Ad/PPSA produced higher transgene expression levels than naked Ad and achieved CAR-independent transduction. Oncolytic Ad (DWP418)/PPSA was able to overcome the nonspecificity of polymer-only therapies by demonstrating cancer-specific killing effects. Furthermore, the DWP418/PPSA nanocomplex elicited a 2.24-fold greater antitumor efficacy than naked Ad in vivo. This was supported by immunohistochemical confirmation of Ad E1As accumulation in MCF7 xenografted tumors. Lastly, intravenous injection of DWP418/PPSA elicited less innate immune response compared to naked Ad, evaluated by interleukin-6 cytokine release into the serum. The increased antitumor effect and improved vector targeting to both CAR-negative and -positive cells make DWP418/PPSA a promising tool for cancer gene therapy. PMID:25400213

Jung, Soo-Jung; Kasala, Dayananda; Choi, Joung-Woo; Lee, Soo-Hwan; Hwang, June Kyu; Kim, Sung Wan; Yun, Chae-Ok



How long do nosocomial pathogens persist on inanimate surfaces? A systematic review  

PubMed Central

Background Inanimate surfaces have often been described as the source for outbreaks of nosocomial infections. The aim of this review is to summarize data on the persistence of different nosocomial pathogens on inanimate surfaces. Methods The literature was systematically reviewed in MedLine without language restrictions. In addition, cited articles in a report were assessed and standard textbooks on the topic were reviewed. All reports with experimental evidence on the duration of persistence of a nosocomial pathogen on any type of surface were included. Results Most gram-positive bacteria, such as Enterococcus spp. (including VRE), Staphylococcus aureus (including MRSA), or Streptococcus pyogenes, survive for months on dry surfaces. Many gram-negative species, such as Acinetobacter spp., Escherichia coli, Klebsiella spp., Pseudomonas aeruginosa, Serratia marcescens, or Shigella spp., can also survive for months. A few others, such as Bordetella pertussis, Haemophilus influenzae, Proteus vulgaris, or Vibrio cholerae, however, persist only for days. Mycobacteria, including Mycobacterium tuberculosis, and spore-forming bacteria, including Clostridium difficile, can also survive for months on surfaces. Candida albicans as the most important nosocomial fungal pathogen can survive up to 4 months on surfaces. Persistence of other yeasts, such as Torulopsis glabrata, was described to be similar (5 months) or shorter (Candida parapsilosis, 14 days). Most viruses from the respiratory tract, such as corona, coxsackie, influenza, SARS or rhino virus, can persist on surfaces for a few days. Viruses from the gastrointestinal tract, such as astrovirus, HAV, polio- or rota virus, persist for approximately 2 months. Blood-borne viruses, such as HBV or HIV, can persist for more than one week. Herpes viruses, such as CMV or HSV type 1 and 2, have been shown to persist from only a few hours up to 7 days. Conclusion The most common nosocomial pathogens may well survive or persist on surfaces for months and can thereby be a continuous source of transmission if no regular preventive surface disinfection is performed. PMID:16914034

Kramer, Axel; Schwebke, Ingeborg; Kampf, Günter



In Vitro Activity of Expanded-Spectrum Pyridazinyl Oxime Ethers Related to Pirodavir: Novel Capsid-Binding Inhibitors with Potent Antipicornavirus Activity  

PubMed Central

Picornaviruses (PV) include human rhinovirus (HRV), the primary cause of the common cold, and the enteroviruses (EV), which cause serious diseases such as poliomyelitis, meningoencephalitis, and systemic neonatal disease. Although no compounds for PV infections have been approved in the United States, pirodavir was one of the most promising capsid-binding compounds to show efficacy in human clinical trials for chemoprophylaxis of the common cold. Susceptibility to hydrolysis precluded its use as an oral agent. We have developed orally bioavailable pyridazinyl oxime ethers that are as potent as pirodavir. Compounds BTA39 and BTA188 inhibited a total of 56 HRV laboratory strains and three clinical isolates as determined by neutral red uptake assay. At concentrations of <100 nM, BTA39 inhibited 69% of the HRV serotypes and isolates evaluated, BTA188 inhibited 75%, and pirodavir inhibited 59% of the serotypes and isolates. The 50% inhibitory concentrations (IC50s) for the two compounds ranged from 0.5 nM to 6,701 nM. The compounds also inhibited EV, including coxsackie A and B viruses (IC50 = 773 to 3,608 nM) and echoviruses (IC50 = 193 to 5,155 nM). BTA39 only inhibited poliovirus strain WM-1 at 204 nM, and BTA188 only inhibited poliovirus strain Chat at 82 nM. EV 71 was inhibited by BTA39 and BTA188, with IC50s of 1 and 82 nM, respectively. Both compounds were relatively nontoxic in actively growing cells (50% cytotoxic doses, ?4,588 nM). These data suggest that these oxime ethers warrant further investigation as potential agents for treating selected PV infections. PMID:15105133

Barnard, D. L.; Hubbard, V. D.; Smee, D. F.; Sidwell, R. W.; Watson, K. G. W.; Tucker, S. P. T.; Reece, P. A. R.



Detection of enteroviruses in untreated and treated drinking water supplies in South Africa.  


Enteric viruses have been detected in many drinking water supplies all over the world. A meaningful number of these supplies were treated and disinfected according to internationally acceptable methods. In addition, counts of bacterial indicators (coliform bacteria and heterotrophic plate count organisms) in these water supplies were within limits generally recommended for treated drinking water and these findings have been supported by epidemiological data on infections associated with drinking water. The shortcomings of conventional treatment methods and indicator organisms to confirm the absence of enteric viruses from drinking water, was generally ascribed to the exceptional resistance of these viruses. In this study, the prevalence of enteroviruses detected from July 2000 to June 2002 in sewage, river-, borehole-, spring- and dam water as well as drinking water supplies treated and disinfected according to international specifications for the production of safe drinking water was analysed. A glass wool adsorption-elution technique was used to recover viruses from 10--20 l of sewage as well as environmental water samples, in the case of drinking water from more than 100 l. Recovered enteroviruses were inoculated onto two cell culture types (BGM and PLC/PRF/5 cells) for amplification of viral RNA with nested-PCR being used to detect the amplified viral RNA. Results from the study demonstrated the presence of enteroviruses in 42.5% of sewage and in 18.7% of treated drinking water samples. Furthermore, enteroviruses were detected in 28.5% of river water, in 26.7% of dam/spring water and in 25.3% of borehole water samples. The high prevalence of coxsackie B viruses found in this study suggested, that a potential health risk and a burden of disease constituted by these viruses might be meaningful. These findings indicated that strategies, other than end-point analysis of treated and disinfected drinking water supplies, may be required to ensure the production of drinking water that does not exceed acceptable health risks. More reliable approaches to ensure acceptable safety of drinking water supplies may be based on control by multiple-barrier principles from catchment to tap using hazard assessment and critical control point (HACCP) principles. PMID:15919105

Ehlers, M M; Grabow, W O K; Pavlov, D N



Treatment with CA-074Me, a Cathepsin B inhibitor, reduces lung interstitial inflammation and fibrosis in a rat model of polymyositis.  


Cathepsin B (CB) is involved in the turnover of proteins and has various roles in maintaining the normal metabolism of cells. In our recent study, CB is increased in the muscles of polymyositis/dermatomyositis (PM/DM). However, the role of CB in interstitial lung disease (ILD) has not been reported. ILD is a frequent complication of PM/DM, which is the leading cause of death in PM/DM. It carries high morbidity and mortality in connective tissue diseases, characterized by an overproduction of inflammatory cytokines and induced fibrosis, resulting in respiratory failure. The etiology and pathogenesis of ILD remain incompletely understood. This study investigated whether treatment with CA-074Me, a specific inhibitor of CB, attenuates ILD in PM. CB expression, inflammation, and fibrosis were analyzed in the lung tissues from patients with PM/DM. The animal model of PM was induced in guinea pigs with Coxsackie virus B1 (CVB1). CA-074Me was given 24?h after CVB1 injection for 7 consecutive days. At the end of the experiment, the animals were killed and lung tissues were collected for the following analysis. Inflammation, fibrosis and apoptosis cells, and cytokines were assessed by histological examinations and immunohistochemical analyses, western blot analysis and transferase-mediated dUTP nick-end labeling assay. In patients with PM/DM, the protein levels of CB were significantly elevated in lung tissues compared with healthy controls, which correlated with increases in inflammation and fibrosis. Similarly, the expression of CB, inflammation and fibrosis, CD8(+) T cell, CD68(+) cell, tumor necrosis factor-alpha, transforming growth factor-beta1 infiltrations, and apoptotic cell death were significantly increased in lung tissues of the guinea-pig model of CVB1-induced PM. These changes were attenuated by the administration of CA-074Me. In conclusion, this study demonstrates that PM/DM increases CB expression in lung tissues and inhibition of CB reduces ILD in a guinea-pig model of CVB1-induced PM. This finding suggests that CB may be a potential therapeutic target for ILD. PMID:25384123

Zhang, Li; Fu, Xiao-Hong; Yu, Yong; Shui, Ruo-Hong; Li, Chun; Zeng, Hai-Ying; Qiao, Yu-Lei; Ni, Li-Yan; Wang, Qiang