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1

Improved adenovirus type 5 vector-mediated transduction of resistant cells by piggybacking on coxsackie B-adenovirus receptor-pseudotyped baculovirus.  

PubMed

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

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

2009-06-01

2

Neonatal coxsackie B virus infection—a treatable disease?  

Microsoft Academic Search

Ten neonates with coxsackie B viral infection presented over a 3-month period. Clinical features included meningoencephalitis, thrombocytopenia, disseminated intravascular coagulopathy, cardiomyopathy, and hepatitis. Eight infants had multiorgan disease, four with severe myocardial dysfunction, of whom two died. All of the infants with severe disease developed symptoms within 7 days of age. In infants presenting within 10 days of birth, in

Penelope A. Bryant; David Tingay; Peter A. Dargaville; Mike Starr; Nigel Curtis

2004-01-01

3

Hepatitis and Encephalitis due to Coxsackie Virus A9 in an Adult.  

PubMed

Coxsackie virus infection most commonly manifests itself in the neonatal period as a multisystem disease. This life-threatening neonatal infection has been recently treated with a new anti-picornaviral drug, pleconaril. In contrast, in adults Coxsackie virus is an uncommon source of hepatitis, but Coxsackie virus type B has been described in case reports to cause hepatitis. This is the first case report of hepatitis and encephalitis secondary to Coxsackie virus type A9 in an adult. This virus was found in a culture of the cerebrospinal fluid and was confirmed by PCR. The patient recovered completely without specific treatment. PMID:22114569

Moreau, Brigitte; Bastedo, Clare; Michel, Rene P; Ghali, Peter

2011-09-01

4

Loss of Coxsackie and adenovirus receptor downregulates ?-catenin expression  

Microsoft Academic Search

Background:The Coxsackie and adenovirus receptor (CAR) has been shown to inhibit cancer cell proliferation, migration, and invasion. The underlying mechanisms, however, are poorly understood.Methods:The differential gene expression in the human colon cancer cell line DLD1 on RNAi-mediated functional CAR knockdown was analysed using oligo-array technology. Expression of ?-catenin was determined by quantitative RT-PCR and western blotting. Proliferation, migration, and invasion

K Stecker; A Koschel; B Wiedenmann; M Anders

2009-01-01

5

Investigation of Potable Water Supply at the Village of Coxsackie, New York.  

National Technical Information Service (NTIS)

The existing Coxsackie Water Treatment Plant is equipped with diatomaceous earth filters for domestic water supply. In an effort to improve the quality of the potable water supplied to their users, the Village of Coxsackie, New York has implemented a pilo...

L. K. Wang C. Gaetani

1986-01-01

6

[Use of the cultural variants of Coxsackie A viruses in virological practice].  

PubMed

Coxsackie A viruses belong to the enteroviruses, the isolation of which from infectious materials and further cultivation are possible only when laboratory animals are infected. The authors could adapt the strains of 17 of 23 serotypes of these viruses to RD cell culture. The strains of 8 serotypes were additionally adapted to Vero cell culture. The cultural variants of Coxsackle A viruses were used to prepare immune sera. The Bacterial and Viral Agents Enterprise, M. P. Chumakov Institute of Poliomyelitis and Virus Encephalitides, Russian Academy of Medical Sciences, has set up the production of bacterial and viral drugs based on the cultural variants of 5 Coxsackie A virus serotypes. The cultural variants of 14 Coxsackie A virus serotypes were used to carry out a virus neutralization test. Examination of more than 600 children from Moscow and the Moscow Region showed the wide circulation of individual Coxsackie A virus serotypes. It also demonstrated a drastic reduction in Coxsackie A-7 virus circulation in the past 50 years. PMID:22834148

Se?bil', V B; Malyshkina, L P; Gracheva, L A; Kozlov, V G

2012-01-01

7

Loss of Coxsackie and adenovirus receptor downregulates ?-catenin expression  

PubMed Central

Background: The Coxsackie and adenovirus receptor (CAR) has been shown to inhibit cancer cell proliferation, migration, and invasion. The underlying mechanisms, however, are poorly understood. Methods: The differential gene expression in the human colon cancer cell line DLD1 on RNAi-mediated functional CAR knockdown was analysed using oligo-array technology. Expression of ?-catenin was determined by quantitative RT-PCR and western blotting. Proliferation, migration, and invasion after CAR knockdown were assessed by in vitro assays, and cell morphology in a three-dimensional context was evaluated using matrigel. Results: Oligo-array technology identified ?-catenin as the strongest downregulated gene after CAR knockdown. Western blotting and quantitative RT-PCR confirmed a reduced ?-catenin expression after CAR knockdown in DLD1 cells and in the rat intestinal cell line IEC-6. Functionally, both cell lines showed a marked increase in proliferation, migration, and invasion on CAR knockdown. In matrigel, both cell lines formed amorphous cell clusters in contrast to well-organised three-dimensional structures of CAR-expressing vector controls. Ectopic ‘re'-expression of ?-catenin in DLD1 and IEC-6 CAR knockdown cells reversed these functional and morphological effects. Conclusion These data suggest that an interaction of CAR and ?-catenin mediates the impact of CAR on cell proliferation, migration, invasion, and morphology.

Stecker, K; Koschel, A; Wiedenmann, B; Anders, M

2009-01-01

8

Isoform-specific expression of the Coxsackie and adenovirus receptor (CAR) in neuromuscular junction and cardiac intercalated discs  

Microsoft Academic Search

BACKGROUND: The Coxsackie and adenovirus receptor (CAR) has a restricted expression pattern in the adult. In skeletal muscle, although CAR is expressed in immature fibers, its transcript levels are barely detectable in mature muscle. This is in contrast to the robust expression observed in the heart. However, both heart and skeletal muscle are susceptible to infection with the Coxsackie B

Christian A Shaw; Paul C Holland; Michael Sinnreich; Carol Allen; Kerstin Sollerbrant; George Karpati; Josephine Nalbantoglu

2004-01-01

9

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

NASA Astrophysics Data System (ADS)

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

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

1984-02-01

10

Expression and function of the coxsackie and adenovirus receptor in Barrett's esophagus and associated neoplasia  

Microsoft Academic Search

Cell surface presence of the coxsackie and adenovirus receptor (CAR) is considered a crucial prerequisite for the uptake of attenuated adenovirus. In cancers, however, a frequent loss of CAR has been noted potentially hampering the success of adenovirus-based therapy. In esophageal Barrett's carcinomas and its precursor lesions CAR presence has not been systematically determined yet. Immunohistochemical assessment in tissue specimens

M Anders; T Rösch; K Küster; I Becker; H Höfler; H J Stein; A Meining; B Wiedenmann; M Sarbia

2009-01-01

11

Severe Coxsackie virus B infection in preterm newborns treated with pleconaril  

Microsoft Academic Search

Four preterm newborn infants with severe multisystem Coxsackie virus B infection were treated with an oral suspension of pleconaril (5 mg\\/kg per day). The patients had myocarditis, fulminant hepatitis, meningoencephalitis and disseminated intravascular coagulopathy. All four infants recovered, and no adverse effects of the treatment were noted. Conclusion: pleconaril needs to be comprehensively evaluated in this population.

Sofia Bauer; Giora Gottesman; Lea Sirota; Ita Litmanovitz; Shay Ashkenazi; Itzhak Levi

2002-01-01

12

Chemical components of Ardisia splendens leaves and their activity against coxsackie A16 viruses.  

PubMed

Using a combination of chromatographic methods, one new flavonol glycoside, myricetin 3,7-di-O-alpha-L-rhamnopyranoside (1), and nine known compounds myricitrin (2), quercetin 3,7-di-O-alpha-L-rhamnopyranoside (3), quercitrin (4), desmanthin-l (5), myricetin 3-O-(3"-O-galloyl)-alpha-L-rhamnopyranoside (6), (+)-catechin (7), benzyl O-1-D-glucopyranoside (8), 2-phenylethyl O-beta-D-glucopyranoside (9), and corilagin (10) were isolated from the leaves of Ardisia splendens Pit. Based on an in vitro test against Coxsackie viruses A16 by SRB assay, only compounds 2, 5, and 10 exhibited activity against Coxsackie viruses A16 with IC50 values of 40.1, 32.2, and 30.5 microM, respectively. This result suggested that compounds 2, 5, and 10 might be potential agents for treating hand, foot and mouth diseases. PMID:25026709

Van Nguyen, Thi Hong; Vien, Trinh Anh; Nhiem, Nguyen Xuan; Van Kiem, Phan; Van Minh, Chau; Long, Pham Quoc; Anh, Luu Tuan; Cuong, Nguyen Manh; Song, Jae-Hyoung; Ko, Hyun-Jeong; Kim, Nanyoung; Park, Seon Ju; Kim, Seung Hyun

2014-05-01

13

Antiviral treatment of Coxsackie B virus infection in human pancreatic islets  

Microsoft Academic Search

Enterovirus infections of the pancreatic islets are believed to trigger or precipitate the near total destruction of ?-cells that constitutes type 1 diabetes (T1D). This study investigated the ability of an anti-picornaviral compound, pleconaril, to block the replication of two ?-cell tropic Coxsackie B4 virus (CBV-4) strains in isolated human islets. The two strains, VD2921 and V89 4557, with demonstrated

Anna-Karin Berg; Annika Olsson; Olle Korsgren; Gun Frisk

2007-01-01

14

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

PubMed

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

2014-01-01

15

DEVELOPMENT OF A RAPID, QUANTITATIVE METHOD FOR THE DETECTION OF INFECTIVE COXSACKIE AND ECHO VIRUSES IN DRINKING WATER  

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...

16

ACQUISITION OF ANTIBODIES TO VARIOUS COXSACKIE AND ECHO VIRUSES AND HEPATITIS A VIRUS IN AGRICULTURAL COMMUNAL SETTLEMENTS IN ISRAEL  

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...

17

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.

2014-01-01

18

Overexpression of coxsackie and adenovirus receptor inhibit growth of human bladder cancer cell in vitro and in vivo  

Microsoft Academic Search

Aim:To study the effect of the overexpression of coxsackie and the adenovirus receptor (CAR) on the growth of the human bladder cancer cell in vitro and in vivo.Methods:A retroviral vector pLXSN-CAR expressing CAR was constructed and confirmed by restriction enzyme mapping. The pLXSN-CAR vector and control vector pLXSN were transfected into the PT67 packaging cell line to generate retrovirus with

Lin-lin Zhang; Da-lin He; Xiang Li; Lei Li; Guo-dong Zhu; Dong Zhang; Xin-yang Wang

2007-01-01

19

A case report of a premature infant with coxsackie B1 meningitis.  

PubMed

This case report describes a 36-week gestational age infant diagnosed with coxsackie B1 meningitis at 20 days of age. A head ultrasound 5 days after diagnosis was consistent with cystic periventricular leukomalacia. The scientific literature does not clearly elucidate differences between bacterial and viral infections in infants. When difficulties arise, it is pertinent to consider a viral etiology for the underlying illness and obtain a detailed maternal and infant history focusing on clinical symptoms, seasonality, geographic location, exposure, and incubation period. Polymerase chain reaction is a rapid and sensitive diagnostic test for the identification of enteroviruses in cerebrospinal fluid, blood, urine, and throat specimens and should be performed as part of the general workup in the evaluation of a febrile infant with sepsis. In retrospect, it may have established an earlier diagnosis of meningitis, consequently preventing the unnecessary use of antibiotics, potentially decreasing the length of hospitalization, and eliminating the need for more detailed investigations to rule out other etiological factors. In addition, treatment with pleconaril may have affected the severity of the encephalitis. This article reviews the pathogenesis, clinical manifestations, and differential diagnoses of enteroviral infections, specifically focusing on the prevention, treatment, and prognosis of the disease and the implications for clinical practice. PMID:18049150

Callen, Jennifer; Paes, Bosco A

2007-10-01

20

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.

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

2012-01-01

21

Antiviral treatment of Coxsackie B virus infection in human pancreatic islets.  

PubMed

Enterovirus infections of the pancreatic islets are believed to trigger or precipitate the near total destruction of beta-cells that constitutes type 1 diabetes (T1D). This study investigated the ability of an anti-picornaviral compound, pleconaril, to block the replication of two beta-cell tropic Coxsackie B4 virus (CBV-4) strains in isolated human islets. The two strains, VD2921 and V89 4557, with demonstrated abilities to cause non-lytic persistence or lytic infection, respectively, in islets, represented two different potential mechanisms behind virus-induced T1D. The virus replication in the islets was studied with and without addition of pleconaril. In addition, islet morphology was studied every day. To test the effects of pleconaril and/or DMSO on the beta-cells' insulin secretion, glucose perifusions were performed on treated and untreated islets. Virus titrations showed a clear reduction of the replication of both strains after pleconaril treatment. The VD2921 strain was inhibited to undetectable levels. The V89 4557 strain, however, showed an initial reduction of titers but virus titers then increased despite the addition of a second dose of pleconaril. This incomplete inhibition of viral replication suggested the existence of a resistant subtype within this strain. Pleconaril treatment reduced the beta-cells' insulin secretion in response to glucose stimulation in some experiments and induced slight morphological changes to the islets compared to untreated controls. In summary, pleconaril reduced the replication of the two beta-cell tropic CBV-4 strains in human islets. However, genetic differences between these strains influenced the effectiveness of pleconaril treatment. This stresses the importance of using multiple viral strains in antiviral tests. PMID:17239967

Berg, Anna-Karin; Olsson, Annika; Korsgren, Olle; Frisk, Gun

2007-04-01

22

Absence of hepatic uptake of Tc-99m sulfur colloid in an infant with Coxsackie B2 viral infection  

SciTech Connect

After the intravenous administration of Tc-99m sulfur colloid, there was found homogeneous lung, renal, and splenic uptake with absence of uptake by the liver and bone marrow of a nine-day-old female infant. More than 20 other doses were dispensed from the same Tc-99m sulfur colloid preparation with the expected biodistribution. A necropsy done two days later showed diffuse hepatic hemorrhagic necrosis without evidence of intravascular fibrin deposition in the lungs or kidneys. The underlying cause of the infant's disease was a Coxsackie B2 viral infection, based upon positive postmortem viral cultures of kidney and liver tissues and characteristic histopathologic lesions of the central nervous system and viscera. The altered biodistribution presumably reflected marked impairment of Kupffer cell function and an apparent increase in pulmonary macrophages.

Hinkle, G.H.; Leonard, J.C.; Krous, H.F.; Alexander, J.B.

1983-06-01

23

[Correction of immune response using purified staphylococcal toxoid and likopid in the secondary immunodeficiency induced by Coxsackie virus B3].  

PubMed

The action of immunomodulators, purified staphylococcal toxoid (PST) and lycopid, on secondary immunodeficiency state developing during infection caused by Coxsackie virus B3 was studied. This defect was manifested by delayed hypersensitivity to sheep red blood cells (SRBC) and the suppression of neutralizing antibodies to poliomyelitis virus. Depending on the scheme of the experiment, PST normalized the defects of immune response to SRBC or poliovirus, increased suppression or showed no activity. Lycopid corrected the defects of humoral response to SRBC. The combination of PST and lycopid was found to produce no increase of suppression. The suggestion was made on the expediency of combination of two (and probably more) immunomodulators for increasing the efficiency of correction of secondary immunodeficiency. PMID:11871300

Semenova, I B; Vasil'eva, I G; Akatov, A K

2001-01-01

24

Cold pressure test producing coronary spasm, coronary thrombosis and myocardial infarction in a patient with IgM antibodies against Coxsackie B virus.  

PubMed

Several lines of evidence have shown that viral infections are capable of causing coronary spasm and precipitating or mimicking clinical myocardial infarction. Here we report the case of a 41-year-old woman with recurrent angina who was admitted to our hospital because of ventricular tachycardia. Laboratory examination revealed positive IgM titers against Coxsackie B virus. Coronary angiography showed normal coronary arteries, but following a cold pressure test severe spasm of all coronaries with thrombotic occlusion of the second marginal branch of the circumflex artery occurred. We conclude that coronary spasm should be clinically suspected in patients with chest pain and ventricular arrhythmia in combination with IgM antibodies against Coxsackie B virus. In these patients, a cold pressure test should be avoided, and antithrombotic and antispastic therapy is recommended. PMID:10844390

Haberbosch, W; Roerich, N; Neuzner, J

1999-01-01

25

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

Microsoft Academic Search

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

Werner Haberbosch; Norbert Roerich; Joerg Neuzner

1999-01-01

26

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.

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

2014-01-01

27

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

PubMed

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

2014-07-01

28

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

PubMed Central

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

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

2002-01-01

29

Distinct different expression of Th17 and Th9 cells in coxsackie virus B3-induced mice viral myocarditis  

PubMed Central

Background Recently, a new subset of CD4+T helper(Th) cell that predominantly secret cytokine interleukin-9(IL-9) is identified, termed Th9 cell. It has been reported to participate in tissue inflammation and autoimmune responses, and induce disease which differed from Th17 cells. Th17 cells have been shown to play a critical role in viral myocarditis (VMC), but whether Th9 cells are involved in the pathogenesis of VMC remains unclear. Results BALB/c mice were intraperitoneally (i.p) injected with coxsackie virus B3(CVB3) for establishing VMC models. Control mice were treated with phosphate-buffered saline i.p. On day 0,7,14,21,28,35,42 after injection, myocardial histopathological changes were evaluated by hematoxylin-eosin staining. Splenic Th17 and Th9 cells subsets were analyzed by flow cytometry. And cardiac IL-17, IL-9 mRNA were measured by semi-quantitative reverse transcription-PCR and nested PCR, respectively. Results showed the levels of Th17 cells and IL-17 mRNA obviously increased in VMC mice on 7 day after infection, peaked on day 28, and highly persisted to at least day 42 (p < 0.05). While the frequencies of Th9 cells and IL-9 mRNA showed no significant difference between VMC and control group throughout the course of the experiment(p > 0.05). Conclusions It was differentiated Th17 but not Th9 cells significantly elevated in the development of CVB3-induced VMC. The microenvironment of VMC seemed to contribute to the differentiation and proliferation of Th17 rather than Th9 cells. Our preliminary data implied Th9 cells could not protect against VMC nor promote the disease.

2011-01-01

30

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.

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

2011-01-01

31

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

PubMed Central

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 (K5, K10 and K15), 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.

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

2011-01-01

32

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: cornelia.halin@pharma.ethz.ch

2009-01-15

33

Relationship between serum insulin autoantibodies, islet cell antibodies and Coxsackie-B4 and mumps virus-specific antibodies at the clinical manifestation of Type 1 (insulin-dependent) diabetes  

Microsoft Academic Search

Summary  In order to elucidate the possible relationship between insulin autoantibodies (IAA), conventional (ICA-IgG) and complement-fixing (CF-ICA) islet cell antibodies and Coxsackie-B4 and mumps virus-specific antibodies (IgG, IgM and IgA classes), we studied 194 children and adolescents with newly diagnosed Type 1 (insulin-dependent) diabetes. Sixty-one (31.4%) of the subjects were IAA-positive at diagnosis and 73.8% (45\\/61) of these also had ICA-IgG

J. Karjalainen; M. Knip; H. Hyöty; P. Leinikki; J. Ilonen; M.-L. Käär; H. K. Åkerblom

1988-01-01

34

Infection of human islets of langerhans with two strains of Coxsackie B virus serotype 1: assessment of virus replication, degree of cell death and induction of genes involved in the innate immunity pathway.  

PubMed

Type 1 diabetes mellitus is believed to be triggered, in part, by one or more environmental factors and human enteroviruses (HEVs) are among the candidates. Therefore, this study has examined whether two strains of HEV may differentially affect the induction of genes involved in pathways leading to the synthesis of islet hormones, chemokines and cytokines in isolated, highly purified, human islets. Isolated, purified human pancreatic islets were infected with strains of Coxsackievirus B1.Viral replication and the degree of CPE/islet dissociation were monitored. The expression of insulin, glucagon, CXCL10, TLR3, IF1H1, CCL5, OAS-1, IFN?, and DDX58 was analyzed. Both strains replicated in islets but only one of strain caused rapid islet dissociation/CPE. Expression of the insulin gene was reduced during infection of islets with either viral strain but the gene encoding glucagon was unaffected. All genes analyzed which are involved in viral sensing and the development of innate immunity were induced by Coxsackie B viruses, with the notable exception of TLR3. There was no qualitative difference in the expression pattern between each strain but the magnitude of the response varied between donors. The lack of virus induced expression of TLR3, together with the differential regulation of IF1H1, OAS1 and IFN?, (each of which has polymorphic variants influence the predisposition to type 1 diabetes), that might result in defective clearance of virus from islet cells. The reduced expression of the insulin gene and the unaffected expression of the gene encoding glucagon by Coxsackie B1 infection is consistent with the preferential ?-cell tropism of the virus. PMID:24249667

Anagandula, Mahesh; Richardson, Sarah J; Oberste, M Steven; Sioofy-Khojine, Amir-Babak; Hyöty, Heikki; Morgan, Noel G; Korsgren, Olle; Frisk, Gun

2014-08-01

35

A One-Step, Triplex, Real-Time RT-PCR Assay for the Simultaneous Detection of Enterovirus 71, Coxsackie A16 and Pan-Enterovirus in a Single Tube  

PubMed Central

The recent, ongoing epidemic of hand, foot, and mouth disease (HFMD), which is caused by enterovirus infection, has affected millions of children and resulted in thousands of deaths in China. Enterovirus 71 (EV71) and coxsackie A16 (CA16) are the two major distinct pathogens for HFMD. However, EV71 is more commonly associated with neurologic complications and even fatalities. Therefore, simultaneously detecting and differentiating EV71 and CA16 specifically from other enteroviruses for diagnosing HFMD is important. Here, we developed a one-step, triplex, real-time RT-PCR assay for the simultaneous detection of EV71, CA16, and pan-enterovirus (EVs) in a single tube with an internal amplification control. The detection results for the serially diluted viruses indicate that the lower limit of detection for this assay is 0.001–0.04 TCID50/ml, 0.02 TCID50/ml, and 0.001 TCID50/ml for EVs, EV71, and CA16, respectively. After evaluating known HFMD virus stocks of 17 strains of 16 different serotypes, this assay showed a favorable detection spectrum and no obvious cross-reactivity. The results for 141 clinical throat swabs from HFMD-suspected patients demonstrated sensitivities of 98.4%, 98.7%, and 100% for EVs, EV71, and CA16, respectively, and 100% specificity for each virus. The application of this one-step, triplex, real-time RT-PCR assay in clinical units will contribute to HFMD surveillance and help to identify causative pathogen in patients with suspected HFMD.

Yan, Qiang; He, Shuizhen; Zhou, Wenbin; Ge, Shengxiang; Xia, Ningshao

2014-01-01

36

Pregnancy outcome following infections by coxsackie, echo, measles, mumps, hepatitis, polio and encephalitis viruses  

Microsoft Academic Search

Women may be infected during pregnancy with infectious agents that are often passed unnoticed; however, the causative agent may still traverse the placenta and infect the developing embryo and fetus. Several of these agents (i.e. rubella, cytomegalovirus or Toxoplasma Gondii) may cause severe fetal damage, but most other infections in pregnancy seem to be much less dangerous to the fetus.

Asher Ornoy; Alexander Tenenbaum

2006-01-01

37

Characterization of a 100-Kilodalton Binding Protein for the Six Serotypes of Coxsackie B Viruses  

Microsoft Academic Search

Viral infection of host cells primarily depends on binding of the virus to a specific cell surface protein. In ordertocharacterizethebindingproteinforgroupBcoxsackieviruses(CVB),detergent-solubilizedmembrane 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

ULLA RAAB DEVERDUGO; HANS-CHRISTOPH SELINKA; MICHAEL HUBER; BORIS KRAMER; JOSEF KELLERMANN; PETER HANS HOFSCHNEIDER; ANDREINHARD KANDOLF

1995-01-01

38

Visceral pathology of acute systemic injury in newborn mice on the onset of Coxsackie virus infection  

PubMed Central

Coxsackievirus B (CVB) is a significant pathogen of neonatal diseases with severe systemic involvement and high mortality. Hence, it is essential to develop a CVB-induced acute systemic disease model on newborn mouse and study the injury at the onset phase. In this work, a clinical strain of CVB3, Nancy, and its variant strain, Macocy, were adopted in 24 hour old neonates by oral infection. The pathological changes in the heart, liver and lung tissues were analyzed by pathology assays. In situ end labeling assay for programmed cell death was carried out for liver tissues. The data on fatality and infection rates and pathology scores were analyzed statistically. The genomic sequences of the two strains were aligned. The model represented the manifest clinical syndromes of hepatitis, pneumonia and myocardial injury at the onset phase, in which massive numbers of hepatocytes had undergone programmed cell death. Statistical and pathological analysis indicated that the myocardial injury was mild, whereas the liver and lung were more severe. The fatality rate, infection and pathology of the two CVB strains were the same. Therefore, two nucleotide mutations in the 5’ UTR and four amino acid mutations in polyprotein, which did not alter virulence, were shown. By peroral CVB infection of neonatal mice, we developed an acute systemic disease model for studying visceral pathology and systemic disease. At the onset of acute neonatal systemic disease, the hepatitis and pneumonia may be the dominant reason of death, as the injury of liver and lung is more severe than that of heart.

Wang, Lulu; Dong, Changyuan; Chen, Dong-E; Song, Zhen

2014-01-01

39

[Polio-like myelitis due to Coxsackie-Virus B 3: Course under treatment with pleconaril].  

PubMed

Enteroviruses are common with infections of the CNS, such as encephalitis and myelitis, but they may cause various diseases in different organ systems, particulary with fatal outcome. Pleconaril is a new orally acting antiviral drug with broad anti-picornavirus activity, which provides to treat rhinoviral and enteroviral infections. To explain the importance for clinical use, we report a case of severe enteroviral infections of the CNS, treated by Pleconaril. A 14 year old girl presented with a severe polio- like myelitis including flaccid paraparesis and urinary incontinence due to Coxsackievirus-infection. Because of prolonged course and virus persistence we treated with Pleconaril, after treatment a remarkable improvement could be noticed, continency and the ability to walk without aid were regained within a few weeks. With the development of new antiviral substances we are now given more opportunities to treat infectious conditions of the central nervous system. We suggest to include enteroviruses in diagnostic procedures since there is an effective treatment with the new drug Pleconaril available. PMID:14520593

Utzig, N; Friedrich, B; Burtzlaff, C; Lauffer, H

2003-01-01

40

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

JEFFREY M. BERGELSON; ANITA KRITHIVAS; LEO CELI; GUSTAVO DROGUETT; MARSHALL S. HORWITZ; THOMAS WICKHAM; RICHARD L. CROWELL

1998-01-01

41

Distinct different expression of Th17 and Th9 cells in coxsackie virus B3-induced mice viral myocarditis  

Microsoft Academic Search

Background  Recently, a new subset of CD4+T helper(Th) cell that predominantly secret cytokine interleukin-9(IL-9) is identified, termed Th9 cell. It has been reported\\u000a to participate in tissue inflammation and autoimmune responses, and induce disease which differed from Th17 cells. Th17 cells\\u000a have been shown to play a critical role in viral myocarditis (VMC), but whether Th9 cells are involved in the

Kong Qing; Wu Weifeng; Yang Fan; Yan Yuluan; Pang Yu; Huang Yanlan

2011-01-01

42

Influence of neopterin and 7,8-dihydroneopterin on the replication of Coxsackie type B5 and influenza A viruses  

Microsoft Academic Search

Pteridine derivatives neopterin and 7,8-dihydroneopterin are produced by human macrophages and dendritic cells upon stimulation\\u000a with interferon-? (IFN-?) and therefore become detectable in increased amounts in humans during cell-mediated (Th1-type) immune\\u000a response. Compounds produced upon influence of cytokine IFN-? often exert antiproliferative and antiviral activity. The aim\\u000a of this study was to investigate the effect of neopterin and 7,8-dihydroneopterin on

Olga Bratslavska; Diana Platace; Edv?ns Miklaševi?s; Dietmar Fuchs; Agris Martinsons

2007-01-01

43

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

PubMed Central

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

2014-01-01

44

Inhibition of Coxsackie B Virus Infection by Soluble Forms of Its Receptors: Binding Affinities, Altered Particle Formation, and Competition with Cellular Receptors  

Microsoft Academic Search

We previously reported that soluble decay-accelerating factor (DAF) and coxsackievirus-adenovirus receptor (CAR) blocked coxsackievirus B3 (CVB3) myocarditis in mice, but only soluble CAR blocked CVB3-mediated pancreatitis. Here, we report that the in vitro mechanisms of viral inhibition by these soluble receptors also differ. Soluble DAF inhibited virus infection through the formation of reversible complexes with CVB3, while binding of soluble

Ian G. Goodfellow; David J. Evans; Anna M. Blom; Dave Kerrigan; J. Scott Miners; B. Paul Morgan; O. Brad Spiller

2005-01-01

45

Autoimmunity and viral infection in diabetes mellitus.  

PubMed Central

Forty-three newly diagnosed insulin-dependent diabetics and thirty control subjects have been examined for cellular hypersensitivity to different pancreatic preparations, rat liver mitochondria and Coxsackie B4 virus. Evidence for cellular hypersensitivity to the pancreatic preparations was shown in the diabetic group, in whom the hypersensitivity to the pancreatic preparations was found to correlate with hypersensitivity to rat liver mitochondria. Hypersensitivity to Coxsackie B4 virus was not found to differ significantly in the diabetic and control groups. Hypersensitivity to liver mitochondria appeared to decrease over a 1-year period. There was an indication that hypersensitivity to Coxsackie B4 was greatest 3 months after diagnosis.

Richens, E R; Ancil, R J

1976-01-01

46

Virus infection and knee injury.  

PubMed Central

Serological evidence of virus infection was sought in 31 consecutive patients presenting with knee swelling and compared with age/sex-matched controls. In a normal age/sex-matched control group, 42% of patients had evidence of recent or past infection with Coxsackie B virus, emphasising the care required in the evaluation of the significance of Coxsackie B neutralization titres in individual patients. Of 12 patients presenting with knee swelling and a history of a twisting injury, eight had serological evidence of recent or past infection with Coxsackie B virus, and one had evidence of a current adenovirus infection.

Driscoll, P; Venner, R; Clements, G B

1987-01-01

47

Hand, foot, and mouth disease on the soles (image)  

MedlinePLUS

Hand, foot, and mouth disease is cause by a coxsackie virus. It produces mouth ulcers and small blisters (vesicles) on the hands and feet. The vesicles often have a reddish border with a white or lighter colored area in ...

48

Hepatitis A and the Vaccine during Pregnancy  

MedlinePLUS

... Hepatitis A or the vaccine while I am breastfeeding? Breastfeeding can continue without interruption if a mother ... 1475- 1476. Ornoy A, Tenenbaum, A. 2006. Pregnancy Outcome following infections by coxsackie, echo, measles, mumps, hepatitis, ...

49

Production of Enterovirus Antisera.  

National Technical Information Service (NTIS)

The production of enterovirus antisera in horses and of rhinovirus antisera in goats is reported. Eight other coxsackie virus, one echo virus, and eight rhinovirus antigens are in progress. Data on inoculations, bleedings, and stored and transferred sera ...

R. W. Brown

1967-01-01

50

Viral Inhibitory Activity of Polyvinylpyrrolidone.  

National Technical Information Service (NTIS)

The investigation was undertaken on the discovery that PVP, in non-toxic concentrations, affords some protection against the cytopathogenic effects in KB cells caused by infection with herpes simplex and Coxsackie B viruses when the cultures are treated w...

B. S. Michaels G. H. Waddell D. D. Zinner M. M. Sigel

1966-01-01

51

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

PubMed

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

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

2012-01-01

52

Chemical synthesis of picornaviral protein primers of RNA replication.  

PubMed

Naturally occurring nucleopeptidic replication primers (VPg-pUpU) of poliovirus and coxsackie virus were chemically synthesized. The synthesis was accomplished via block-coupling of two minimally protected fragments of the target structures: a short RNA-nucleopeptide and a longer peptide segment containing diverse side-chain functionalities. The synthetic VPg-pUpU of coxsackie virus was characterized by NMR spectroscopy. PMID:16990933

Kriek, Nicole M A J; Meeuwenoord, Nico J; van den Elst, Hans; Heus, Hans A; van der Marel, Gijsbert A; Filippov, Dmitri V

2006-10-01

53

Masticator myopathy.  

PubMed

A 31-year-old woman developed low-grade fever and pain and swelling of the masticatory muscles. A T2-weighted magnetic resonance image showed high signal intensity in these muscles. Coxsackie B3 and echo 30 viruses were detected from a nasopharyngeal swab and feces, respectively. The clinical symptoms accompanied a marked decline in the serum immunoglobulin G level with progressive eosinophilia. Her symptoms disappeared by 8 weeks after onset. She was diagnosed as having masticator myopathy, which has rarely been reported in humans. The present case suggests that masticator myopathy is associated with coxsackie or echo virus infection. PMID:12811784

Yamada, Shigeo; Ogawa, Tomoko; Nishimiya, Jin; Yuasa, Tatsuhiko; Taketazu, Fumitoshi

2003-07-01

54

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

Microsoft Academic Search

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

Joseph T. C. Shieh; Jeffrey M. Bergelson

2002-01-01

55

Efficient targeting of adenoviral vectors to integrin positive vascular cells utilizing a CAR-cyclic RGD linker protein  

Microsoft Academic Search

Vascular smooth muscle (VSMC) and endothelial cells (EC) are particularly resistant to infection by type 5 adenovirus (Ad) vectors. To overcome this limitation and target Ad vectors to ubiquitously expressed ?V?3\\/5 integrins, we have generated a linker protein consisting of the extracellular domain of the coxsackie adenovirus receptor (CAR) connected via avidin to a biotinylated cyclic (c) RGD peptide. After

Y. D. Krom; J. C. E. Gras; R. R. Frants; L. M. Havekes; T. J. van Berkel; E. A. L. Biessen; K. Willems van Dijk

2005-01-01

56

Molecular evolution of swine vesicular disease virus  

Microsoft Academic Search

Phylogenetic analysis was used to examine the evolutionary relationships within a group of coxsackie B viruses that contained representatives of the major serotypes of this group and 45 isolates of swine vesicular disease virus (SVDV) from Asia and Europe. Separate analyses of sequence data from two regions of the viral genomes encoding the VP1 and 3BC genes both revealed that

Gang Zhang; Daniel T. Haydon; Nick J. Knowles; John W. McCauley

1999-01-01

57

Enhanced antitumor effect of RGD fiber-modified adenovirus for gene therapy of oral cancer  

Microsoft Academic Search

Current clinical success rates of adenoviral vector (Adv)–based gene therapy of squamous cell carcinoma (SCC) of the head and neck remain unsatisfactory. A major problem with this approach is thought to be related to low Adv transduction efficiency due to weak expression of the adenovirus receptor, coxsackie–adenovirus receptor (CAR), in SCC. To improve the limited infectivity of Adv in oral

Hironari Dehari; Yoshinori Ito; Takafumi Nakamura; Masayoshi Kobune; Katsunori Sasaki; Noriyuki Yonekura; Geniku Kohama; Hirofumi Hamada

2003-01-01

58

Infections and autoimmune diseases  

Microsoft Academic Search

The high percentage of disease-discordant pairs of monozygotic twins demonstrates the central role of environmental factors in the etiology of autoimmune diseases. Efforts were first focussed on the search for triggering factors. The study of animal models has clearly shown that infections may trigger autoimmune diseases, as in the case of Coxsackie B4 virus in type I diabetes and the

Jean-François Bach

2005-01-01

59

Glycocalyx Restricts Adenoviral Vector Access to Apical Receptors Expressed on Respiratory Epithelium In Vitro and In Vivo: Role for Tethered Mucins as Barriers to Lumenal Infection  

Microsoft Academic Search

Inefficient adenoviral vector (AdV)-mediated gene transfer to the ciliated respiratory epithelium has hin- dered gene transfer strategies for the treatment of cystic fibrosis lung disease. In part, the inefficiency is due to an absence of the coxsackie B and adenovirus type 2 and 5 receptor (CAR) from the apical membranes of polarized epithelia. In this study, using an in vitro

Jaclyn R. Stonebraker; Danielle Wagner; Robert W. Lefensty; Kimberlie Burns; Sandra J. Gendler; Jeffrey M. Bergelson; Richard C. Boucher; Wanda K. O'Neal; Raymond J. Pickles

2004-01-01

60

Epidemiological, virological, and clinical features of an epidemic of hand, foot, and mouth disease in England and Wales  

Microsoft Academic Search

Summary We describe the epidemiological, virological, and clinical features of an epidemic of hand, foot, and mouth disease, attributed to coxsackie A virus serotype 16, that occurred throughout England and Wales in the last quarter of 1994. Nine hundred and fifty-two cases were reported by spotter practices that make weekly returns to the Royal College of General Practitioners, which made

J W A Bendig; D M Fleming; C K Fairley; D Owen; R C Matthews; E Miller

61

Detection of Coxsackievirus B3 RNA in myocardial tissues by the polymerase chain reaction.  

PubMed Central

Coxsackievirus B3 is a possible etiologic agent in some forms of myocarditis and idiopathic dilated cardiomyopathy. A method for the detection of coxsackievirus B3 RNA was developed using the polymerase chain reaction based on the amplification of a cDNA copy of the positive-strand viral RNA. The fidelity of the method was established in two murine models for coxsackie B3 myocarditis. All cardiac specimens with adequate RNA for study from coxsackie B3-infected mice contained detectable viral RNA, in contrast to none of control specimens from noninfected mice. The sensitivity of the technique was established at approximately 1 to 100 plaque-forming units of virus per gram of tissue, and the specificity was established as limited to the coxsackievirus B3 serotype among nine viruses tested. In patients with myocarditis, one of five specimens contained detectable viral RNA, whereas none of 11 specimens from patients with idiopathic dilated cardiomyopathy or 21 myocardial specimens from patients with a wide variety of other cardiac disorders contained detectable coxsackie B3 viral RNA. The results show that the polymerase chain reaction is a useful means for detecting coxsackie viral RNA and its application should help in the evaluation of hypotheses concerning the infectious etiology of human myocarditis and idiopathic dilated cardiomyopathy. Images Figure 2 Figure 3 Figure 4 Figure 5

Weiss, L. M.; Movahed, L. A.; Billingham, M. E.; Cleary, M. L.

1991-01-01

62

IDENTIFICATION AND DETECTION OF WATER-BORNE VIRUSES BY IMMUNOENZYMATIC METHODS  

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...

63

Membrane Adsorption with Direct Cell Culture Combined with Reverse Transcription-PCR as a Fast Method for Identifying Enteroviruses from Sewage  

PubMed Central

We present a new approach for the detection and identification of enteroviruses concentrated and isolated from sewage. Samples were collected from two study sites located at Nicosia and Limassol sewage treatment plants in Cyprus. Viruses were adsorbed to cellulose nitrate membrane filters, cultured directly from the membrane filters by using the VIRADEN method, and identified by reverse transcription-PCR, followed by 5? untranslated region (5?-UTR) restriction fragment length polymorphism (RFLP) analysis and partial sequencing of the VP1 protein coding region. Initial subgrouping based on the HpaII restriction profile showed that all of the isolates except one belonged to the same genetic subcluster. Partial VP1 sequencing revealed that most isolates belonged to serotypes coxsackie B4 (42.5%) and coxsackie ?9 (30%), whereas coxsackie B2 (17.5%) and coxsackie B1 (3%) isolates were less frequently observed. One poliovirus type 2 isolate (2.5%) of vaccine origin was also found. The HpaII digests predicted the genetic subcluster for all isolates. They also accurately differentiated the isolates as nonpolio or polio isolates. This approach seems to be very promising for environmental surveillance of enterovirus circulation and epidemiology, with all of the significant effects that this entails for public health. Partial VP1 sequencing is efficient for molecular serotyping of enteroviruses, while 5?-UTR RFLP analysis with HpaII can also be considered an asset for the initial subclassification of enterovirus isolates.

Papaventsis, D.; Siafakas, N.; Markoulatos, P.; Papageorgiou, G. T.; Kourtis, C.; Chatzichristou, E.; Economou, C.; Levidiotou, S.

2005-01-01

64

The role of coxsackieviruses infection in the children of insulin dependent diabetes mellitus.  

PubMed

In the present study, there were two groups of diabetic patients. The first group was newly diagnosed diabetic patients of less than one year duration of disease. They were 40 patients. The second group was diabetic patients with more than one year duration of disease. They were 30 patients. The control group was 30 normal healthy children. Evidence of virus infection was detected by tissue culture isolation, neutralization test, RT-PCR, IgM, and IgG specific antibodies for coxsackie B viruses. There was significant increase in percent of tissue culture isolation of EV in group I more than group II while the percent in the two groups were significantly increased than the control group. Identification of the type of EV by neutralization test revealed that most of the type cases were Coxsackie B4 virus and one case was Coxsackie B6 virus (the most important diabetogenic strain). Viral RNA detection by PCR was done and revealed that most of cases in group I diabetic patients were positive for enterovirus while one case in group II of diabetic patients was positive. As regards Coxsackie virus B IgM antibodies positivity, there was increase in the percent in group I than group II, as it is a marker of acute infection. As regards Coxsackie virus B IgG antibodies positivity, there was no significant difference between group I and group II of diabetic patients, as it is a marker of past infection and it persists for years after the first episode. PMID:17265619

Maha, M M; Ali, M A; Abdel-Rehim, Soad E; Abu-Shady, E A; El-Naggar, B M; Maha, Y Z

2003-01-01

65

CAR regulates epithelial cell junction stability through control of E-cadherin trafficking  

PubMed Central

CAR (Coxsackie and Adenovirus Receptor) is the primary docking receptor for typeB coxsackie viruses and subgroup C adenoviruses. CAR is a member of the JAM family of adhesion receptors and is located to both tight and adherens junctions between epithelial cells where it can assemble adhesive contacts through homodimerisation in trans. However, the role of CAR in controlling epithelial junction dynamics remains poorly understood. Here we demonstrate that levels of CAR in human epithelial cells play a key role in determining epithelial cell adhesion through control of E-cadherin stability at cell-cell junctions. Mechanistically, we show that CAR is phosphorylated within the C-terminus by PKC? and that this in turn controls Src-dependent endocytosis of E-cadherin at cell junctions. This data demonstrates a novel role for CAR in regulating epithelial homeostasis.

Morton, Penny E.; Hicks, Alexander; Nastos, Theodoros; Santis, George; Parsons, Maddy

2013-01-01

66

Hand, foot and mouth disease in Nagpur.  

PubMed

Hand, foot and mouth disease (HFMD) is a viral infection of children caused by Coxsackie virus-A16, a type of enterovirus closely related with the virus that causes herpangina. Although seen worldwide, it is not common in India. Hand, foot and mouth disease is sporadically reported from India as a mild illness. This report describes four cases of HFMD from Nagpur, Central India, seen between September 2005 and April 2006. All patients presented with a mild febrile prodrome followed by the appearance of aphthous-like oral ulcers and vesicular lesions on the hands and feet. All cases were clinically diagnosed. Coxsackie virus A16 was isolated from the serum of one of the patients. All the patients were in the age group of 3-5 years from different schools. It was a mild illness and all the four patients recovered without any complication. There were no secondary cases in the families. PMID:18388372

Saoji, Vikrant A

2008-01-01

67

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

1997-01-01

68

[The use of the natural mineral saponite for water decontamination].  

PubMed

Sorption properties were studied of natural and activated specimens of saponite with respect to poliomyelitis virus, Coxsackie B 1 and B 6 viruses as well as to Enterobacteriaceae group bacteria. With the purpose of comparing the processes of sorption of microorganisms, other minerals were also used, such as bentonite, alunite, glauconite, ceolite. The natural saponite adsorptive properties were found out to undergo changes during the process of thermoactivation. Mechanisms are discussed of a decontaminating effect of thermoactivated saponite in water. PMID:10423991

Hyrin, V M; Bo?ko, I I; Rudychenko, V F

1999-01-01

69

Adenoviral Gene Vector Tethering to Nanoparticle Surfaces Results in Receptor-Independent Cell Entry and Increased Transgene Expression  

Microsoft Academic Search

The present studies investigated the hypothesis that affinity immobilization of replication-defective adenoviruses (Ad) on the surfaces of biodegradable nanoparticles (NP) can improve transduction through uncoupling cellular uptake from the coxsackie–adenovirus receptor (CAR). Ad was tethered to the surfaces of polylactide-based NP that were surface-activated using a photoreactive polyallylamine–benzophenone–pyridyldithiocarboxylate polymer, which enabled (via thiol chemistry) the covalent attachment of Ad-binding proteins,

Michael Chorny; Ilia Fishbein; Ivan S. Alferiev; Origene Nyanguile; Richard Gaster; Robert J. Levy

2006-01-01

70

Persistence of enteroviral RNA in chronic fatigue syndrome is associated with the abnormal production of equal amounts of positive and negative strands of enteroviral RNA  

Microsoft Academic Search

A subgenomic restriction fragment from cDNA pre- pared from Coxsackie B2 virus (CVB2) RNA was snbcloned into a riboprobe vector allowing the produc- tion of enteroviral group-specific RNA probes comple- mentary to either the positive (genomic) or negative (template) strand of enteroviral RNA. These ribo- probes were used to follow productive infection of cultured cells by CVB2; as expected, positive

Louise Cunningham; N. E. Bowles; V. Dubowitz; L. C. Archard

1990-01-01

71

Chapter 7: Mumps  

Microsoft Academic Search

symptoms, particularly among children less than 5 years.2,3 Not all cases of parotitis--especially sporadic ones--are due to mumps infection. Parotitis can also be caused by parainfluenza virus types 1 and 3, influenza A virus, Coxsackie A virus, echovirus, lymphocytic choriomeningitis virus, human immunodeficiency virus, and other non-infectious causes such as drugs, tumors, immunologic diseases, and obstruction of the salivary duct.

W. William Schluter; Melinda Wharton

72

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

PubMed Central

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

Garg, Geeta; Thami, Gurvinder P

2012-01-01

73

HIV-related opsoclonus–myoclonus–ataxia syndrome: Report on two cases  

Microsoft Academic Search

Opsoclonus–myoclonus–ataxia (OMA) syndrome is a rare neurological disorder, characterized by a rapid onset of generalized myoclonus in association with chaotic multi-directional eye movements and, less frequently, cerebellar ataxia. OMA is commonly related to a paraneoplastic process, specifically neuroblastoma in children and lung or breast cancer in adults. Nevertheless, OMA may occur in association with various infectious agents, such as Coxsackie

Natlada Kanjanasut; Kammant Phanthumchinda; Roongroj Bhidayasiri

2010-01-01

74

Genetic incorporation of the protein transduction domain of Tat into Ad5 fiber enhances gene transfer efficacy  

Microsoft Academic Search

BACKGROUND: Human adenovirus serotype 5 (Ad5) has been widely explored as a gene delivery vector for a variety of diseases. Many target cells, however, express low levels of Ad5 native receptor, the Coxsackie-Adenovirus Receptor (CAR), and thus are resistant to Ad5 infection. The Protein Transduction Domain of the HIV Tat protein, namely PTDtat, has been shown to mediate protein transduction

Tie Han; Yizhe Tang; Hideyo Ugai; Leslie E Perry; Gene P Siegal; Juan L Contreras; Hongju Wu

2007-01-01

75

ISOLATION OF ENTERIC VIRUSES IN ONTARIO DURING 1960-1962.  

PubMed

During the past three years at the Central Laboratory, Ontario Department of Health, 681 isolations were made in tissue culture from 6822 specimens submitted for virus studies by physicians and hospitals from all over Ontario. Nearly 74% of the isolates were enteroviruses, approximately 5% adenoviruses and about 1% reoviruses. The remaining 20% are still to be identified.Although the bulk of isolations was made during the same three-month period (August, September and October) of each year, the predominant virus types varied from year to year. Poliovirus 1 was most commonly encountered in 1960, Coxsackie B5 in 1961 and Echo 9 in 1962.Among other types isolated in smaller numbers were Coxsackie A1, 9 and 10, Coxsackie B1, 2, 3 and 4, Echo 1, 2, 5, 6, 7, 8, 11, 14, 17, 18 and 19, Reovirus 1, 2 and 3, Adenovirus 1, 2, 3, 4, 5, 7 and 16, as well as Frater-type virus. Most of these types were isolated for the first time in Ontario and represent additions to the existing list of viruses known to occur in this province. PMID:14052975

KELEN, A E; BELBIN, D; LESIAK, J M; LABZOFFSKY, N A

1963-11-01

76

Transductional targeting of adenovirus vectors for gene therapy  

PubMed Central

Cancer gene therapy approaches will derive considerable benefit from adenovirus (Ad) vectors capable of self-directed localization to neoplastic disease or immunomodulatory targets in vivo. The ablation of native Ad tropism coupled with active targeting modalities has demonstrated that innate gene delivery efficiency may be retained while circumventing Ad dependence on its primary cellular receptor, the coxsackie and Ad receptor. Herein, we describe advances in Ad targeting that are predicated on a fundamental understanding of vector/cell interplay. Further, we propose strategies by which existing paradigms, such as nanotechnology, may be combined with Ad vectors to form advanced delivery vehicles with multiple functions.

Glasgow, JN; Everts, M; Curiel, DT

2007-01-01

77

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

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

2000-01-01

78

Antiviral spirooliganones A and B with unprecedented skeletons from the roots of Illicium oligandrum.  

PubMed

Two novel spirooliganones A (1) and (2), a pair of spiro carbon epimers, with a rare dioxaspiro skeleton were isolated from the roots of Illicium oligandrum. The structures were fully determined by spectroscopic analysis and chemical methods, especially modified Mosher's method, and X-ray diffraction analysis. Spirooliganone B was found to exhibit more potent activities against coxsackie virus B3 and influenza virus A (H3N2) (IC50 3.70-5.05 ?M) than spirooliganone A. The biosynthetic pathway involving a hetero-Diels-Alder reaction of the epimers was proposed. PMID:23937631

Ma, Shuang-Gang; Gao, Rong-Mei; Li, Yu-Huan; Jiang, Jian-Dong; Gong, Ning-Bo; Li, Li; Lü, Yang; Tang, Wen-Zhao; Liu, Yun-Bao; Qu, Jing; Lü, Hai-Ning; Li, Yong; Yu, Shi-Shan

2013-09-01

79

Evaluation of the anti-microbial and anti-inflammatory activities of the medicinal plants Dodonaea viscosa, Rumex nervosus and Rumex abyssinicus.  

PubMed

The crude extracts of the leaves of Dodonaea viscosa and Rumex nervosus as well as of the root of Rumex abyssinicus were tested for anti-microbial and anti-inflammatory activities. It was observed that the three plants possess antibacterial activity against Streptococcus pyogenes and Staphylococcus aureus and strong activity against Coxsackie virus B3 and influenza A virus. In contrast, none of them exhibited anti-fungal activity. The anti-inflammatory activity test results verified that only R. abyssinicus inhibited the synthesis of prostaglandin (PG) E(2). PMID:12628410

Getie, M; Gebre-Mariam, T; Rietz, R; Höhne, C; Huschka, C; Schmidtke, M; Abate, A; Neubert, R H H

2003-02-01

80

Adenoviruses isolated from wild gorillas are closely related to human species C viruses.  

PubMed

We have isolated and cultured three distinct adenoviruses from wild gorillas. Phylogenetic analysis grouped the viruses with human adenovirus species C based on DNA polymerase, hexon, and E4ORF6 genes. The three wild gorilla adenoviruses clustered with the other species C captive gorilla adenoviruses, forming a branch separate from human and chimpanzee/bonobo adenoviruses. Animal sera to the three newly isolated viruses did not cross-neutralize, demonstrating serological distinctiveness. The human adenovirus 5 fiber knob blocked infection, suggesting use of the Coxsackie and Adenovirus Receptor. These viruses may provide viral vectors with properties distinct from chimpanzee adenovirus and human adenovirus vectors. PMID:23806387

Duncan, McVey; Cranfield, Michael R; Torano, Holly; Kuete, Hubert M; Lee, Grace P; Glenn, Andrew; Bruder, Joseph T; Rangel, David; Brough, Douglas E; Gall, Jason G

2013-09-01

81

High Frequency of Human Enterovirus Species C Circulation in Madagascar  

PubMed Central

Four poliomyelitis outbreaks caused by vaccine-derived polioviruses have been reported recently, including one in Madagascar in 2002. In all cases, the viral strains involved were recombinant between poliovirus vaccine strains and nonpoliovirus strains, probably enterovirus species C. Nevertheless, little is known about the circulation and epidemiology of enteroviruses in the regions where these outbreaks occurred. To assess the circulation of enteroviruses (particularly enterovirus species C) in Madagascar, we genetically characterized 55 enterovirus strains isolated between 1994 and 2002. The strains were identified and compared by partially sequencing the region encoding the VP1 capsid protein. Phylogenetic analysis and pairwise comparison with prototype enterovirus strains distinguished two different species: 25 isolates belonged to human enterovirus B species, and 30 isolates were identified as coxsackievirus A13, A15, A17, A18, A20, A21, and A24, belonging to the human enterovirus species C. The relatively high frequency and the wide distribution of species C coxsackie A viruses in different regions of Madagascar suggest that they had been silently and widely circulating in the country during the whole study period. The circulation of coxsackie A viruses, combined with the low routine oral polio vaccine coverage, may have played a role in the emergence of the recent outbreak in Madagascar.

Rakoto-Andrianarivelo, Mala; Rousset, Dominique; Razafindratsimandresy, Richter; Chevaliez, Stephane; Guillot, Sophie; Balanant, Jean; Delpeyroux, Francis

2005-01-01

82

Second-step concentration of viruses in drinking and surface waters using polyethylene glycol hydroextraction.  

PubMed

In our laboratory, virus adsorbed to talc--Celite layers is eluted with 100 mL of 10% fetal calf serum (FCS) in normal saline (pH 9.0). A further 10-fold reduction in the volume of the eluate was necessary before its inoculation into cell cultures. A 100-mL volume of an experimentally contaminated sample was placed in a dialysis sac and hydroextracted overnight (4 degrees C) with polyethylene glycol (PEG) 6000. The viscous material remaining in the sac was resuspended in 10 mL of Earle's balanced salt solution. After membrane filtration (0.2 micron), the concentrate was plaque assayed in BS-C-1 cells. Using this technique, recoveries of five laboratory-adapted enteric viruses (polio 1, echo 6, coxsackie B5, coxackie A9, and reo 3) and four freshly isolated enteric virus strains (polio 1, echo 1, coxsackie B3, and reo) ranged from 87 to 97%. In comparative tests, PEG hydroextraction was simpler and superior to organic flocculation. PMID:224994

Ramia, S; Sattar, S A

1979-05-01

83

Sesquiterpenes from the roots of Illicium jiadifengpi.  

PubMed

Two new sesquiterpenes (1, 2) and two new sesquiterpene glycosides (3, 4) with a seco-prezizaane skeleton, three new allo-cedrane sesquiterpene glycosides (5-7), and a new acorane sesquiterpene (8) as well as 15 known analogues were isolated from the roots of Illicium jiadifengpi used for the treatment of rheumatoid arthritis. The structures of these compounds were elucidated by extensive spectroscopic analysis and chemical methods. The absolute configurations of compounds 5-7 were confirmed by CD experiments. The configuration of compound 8 was assigned by single-crystal X-ray crystallographic analysis and CD experiments. Compounds 4, 6, 7, 14, and 23 showed moderate antiviral activities against Coxsackie virus B3, and all compounds were inactive when evaluated for their cytotoxic activities against five human tumor cell lines and neuroprotection. PMID:23877924

Zhang, Guijie; Zhuang, Pengyu; Wang, Xiaojing; Yu, Shishan; Ma, Shuanggang; Qu, Jing; Li, Yong; Liu, Yunbao; Zhang, Yan; Yu, Dequan

2013-08-01

84

Isolation of marine bacteria with antiviral properties.  

PubMed

We report in this study the isolation of marine bacteria with antiviral properties that have been tentatively classified as Moraxella. These bacteria retained their virucidal capacity after prolonged subcultivation in the laboratory. The virus-inactivating agent could not be separated from the viable marine bacteria, indicating that the active agent(s) either remains associated to the microorganisms or has a very short lifetime, or both. The antiviral capacity of the isolated microorganisms was highly specific for poliovirus. No virucidal effect was observed against other strains of enteroviruses, such as Coxsackie and ECHO virus, rotavirus SA11, or bacteriophages proposed as indicators of the virological quality of water, such as coliphage f2 and bacteriophage B40-8, which infects Bacteroides fragilis. PMID:2558789

Girones, R; Jofre, J T; Bosch, A

1989-11-01

85

Human enterovirus surveillance in the Slovak Republic from 2001 to 2011.  

PubMed

We report the outcome of an 11-year programme monitoring sewage water and acute flaccid paralysis (AFP) cases as part of the World Health Organization (WHO) strategy for polio eradication in the Slovak Republic (SR). Polioviruses (PV) and non-polio enteroviruses (NPEV), prior to and after the change in polio vaccination strategy, were detected. Sewage treatment plant samples from 48 localities spread over the Western, Central and Eastern regions and clinical material from AFP cases were examined. The WHO standard procedures were followed with regard to virus isolation and identification. There were 538 commonly detected human enteroviruses (HEVs) including 213 (40%) coxsackie B viruses (CBV), 200 (37%) echoviruses and 113 (21%) Sabin-like PVs (PV1, 2, 3) including vaccine-derived poliovirus (VDPV) isolates. The percentage of PV isolates fell from 66% to 30% during 2001-2005 and thereafter fell to zero. CBV5, CBV2 and echovirus 3 were the NPEVs endemic during the study period. PMID:23507533

Klement, C; Kissova, R; Lengyelova, V; Stipalova, D; Sobotova, Z; Galama, J M D; Bopegamage, S

2013-12-01

86

The molecular interaction of CAR and JAML recruits the central cell signal transducer PI3K.  

PubMed

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 alphabeta 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. PMID:20813955

Verdino, Petra; Witherden, Deborah A; Havran, Wendy L; Wilson, Ian A

2010-09-01

87

[Herpangina and erythema multiforme in a three-year boy].  

PubMed

The dentist can be confronted with a vesiculobullous lesion of the oral mucosa are a symptoms of herpes infection (herpangina) of throat. Human enteroviruses (HEVs) are a major cause of herpangina. Herpangina is an acute viral infection caused by certain viruses Coxsackie, is spread by respiratory droplets. The infection is mainly encountered in young children. Oral lesions rarely more than 7 days; treatment is symptomatic. Viral throat infections may accompany various skin rashes, such as erythema multiforme (which can also occur without any connection with a viral infection). At work was presented a case of 3-year-old boy with herpes symptoms of sore throat and mild forms of erythema multiforme. PMID:24455841

Brzezi?ski, Piotr

2013-01-01

88

Borna disease virus nucleoprotein inhibits type I interferon induction through the interferon regulatory factor 7 pathway.  

PubMed

The expression of type I interferon (IFN) is one of the most potent innate defences against viral infection in higher vertebrates. Borna disease virus (BDV) establishes persistent, noncytolytic infections in animals and in cultured cells. Early studies have shown that the BDV phosphoprotein can inhibit the activation of type I IFN through the TBK1-IRF3 pathway. The function of the BDV nucleoprotein in the inhibition of IFN activity is not yet clear. In this study, we demonstrated IRF7 activation and increased IFN-?/? expression in a BDV-persistently infected human oligodendroglia cell line following RNA interference-mediated BDV nucleoprotein silencing. Furthermore, we showed that BDV nucleoprotein prevented the nuclear localisation of IRF7 and inhibited endogenous IFN induction by poly(I:C), coxsackie virus B3 and IFN-?. Our findings provide evidence for a previously undescribed mechanism by which the BDV nucleoprotein inhibits type I IFN expression by interfering with the IRF7 pathway. PMID:23939047

Song, Wuqi; Kao, Wenping; Zhai, Aixia; Qian, Jun; Li, Yujun; Zhang, Qingmeng; Zhao, Hong; Hu, Yunlong; Li, Hui; Zhang, Fengmin

2013-09-01

89

Detection of viral pathogens by reverse transcriptase PCR and of microbial indicators by standard methods in the canals of the Florida Keys.  

PubMed

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, D W; Gibson, C J; Lipp, E K; Riley, K; Paul, J H; Rose, J B

1999-09-01

90

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

PubMed Central

Between October 2001 and April 2002, five cases of acute flaccid paralysis (AFP) associated with type 2 vaccine-derived polioviruses (VDPVs) were reported in the southern province of the Republic of Madagascar. To determine viral factors that favor the emergence of these pathogenic VDPVs, we analyzed in detail their genomic and phenotypic characteristics and compared them with co-circulating enteroviruses. These VDPVs appeared to belong to two independent recombinant lineages with sequences from the type 2 strain of the oral poliovaccine (OPV) in the 5?-half of the genome and sequences derived from unidentified species C enteroviruses (HEV-C) in the 3?-half. VDPV strains showed characteristics similar to those of wild neurovirulent viruses including neurovirulence in poliovirus-receptor transgenic mice. We looked for other VDPVs and for circulating enteroviruses in 316 stools collected from healthy children living in the small area where most of the AFP cases occurred. We found vaccine PVs, two VDPVs similar to those found in AFP cases, some echoviruses, and above all, many serotypes of coxsackie A viruses belonging to HEV-C, with substantial genetic diversity. Several coxsackie viruses A17 and A13 carried nucleotide sequences closely related to the 2C and the 3Dpol coding regions of the VDPVs, respectively. There was also evidence of multiple genetic recombination events among the HEV-C resulting in numerous recombinant genotypes. This indicates that co-circulation of HEV-C and OPV strains is associated with evolution by recombination, resulting in unexpectedly extensive viral diversity in small human populations in some tropical regions. This probably contributed to the emergence of recombinant VDPVs. These findings give further insight into viral ecosystems and the evolutionary processes that shape viral biodiversity.

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

2007-01-01

91

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.

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

1999-01-01

92

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

PubMed Central

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

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

2011-01-01

93

Improved detection of gastrointestinal pathogens using generalised sample processing and amplification panels.  

PubMed

We aimed to streamline the diagnosis of gastrointestinal disease by producing multiplexed real time polymerase chain reaction (PCR) panels employing universal sample processing for DNA and RNA containing pathogens. A total of 487 stored, previously characterised stool samples comprising bacterial, viral, protozoan and Clostridium difficile positive samples were tested using four multiplexed real time PCR panels. A further 81 pre-selected clinical samples from a teaching hospital were included to provide an independent validation of assay performance. Improved sensitivity was achieved using the protozoan panels and 16 more mixed infections were observed compared to tests with conventional methods. Using the C. difficile panels, 100% sensitivity was achieved when compared to the gold standard of toxigenic culture. In addition, hypervirulent strains including ribotype 027 could be identified directly from primary sample without the need for ribotyping methods. Bacterial and viral panels detecting Salmonella, Shigella, Campylobacter, Yersinia enterocolitica, Listeria monocytogenes, norovirus groups I and II, rotavirus A, astrovirus, sapovirus, rotavirus B, adenovirus and adenovirus 40/41 performed as well as conventional methods, whilst allowing detection in 3 hours from processing to result. Multiplex real time PCR panels with universal sample preparation allow streamlined, rapid diagnosis of gastrointestinal pathogens whilst extending the characterisation of pathogens present in stool samples from affected patients. PMID:24300711

Siah, Shoo Peng; Merif, Juan; Kaur, Kiran; Nair, Jiny; Huntington, Peter G; Karagiannis, Thomas; Stark, Damien; Rawlinson, William; Olma, Tom; Thomas, Lee; Melki, John R; Millar, Douglas S

2014-01-01

94

Viruses associated with influenza-like-illnesses in Papua New Guinea, 2010.  

PubMed

Influenza-like-illness can be caused by a wide range of respiratory viruses. The etiology of influenza-like-illness in developing countries such as Papua New Guinea is poorly understood. The etiological agents associated with influenza-like-illness were investigated retrospectively for 300 nasopharyngeal swabs received by the Papua New Guinea National Influenza Centre in 2010. Real-time PCR/RT-PCR methods were used for the detection of 13 respiratory viruses. Patients with influenza-like-illness were identified according to the World Health Organization case definition: sudden onset of fever (>38°C), with cough and/or sore throat, in the absence of other diagnoses. At least one viral respiratory pathogen was detected in 66.3% of the samples tested. Rhinoviruses (17.0%), influenza A (16.7%), and influenza B (12.7%) were the pathogens detected most frequently. Children <5 years of age presented with a significantly higher rate of at least one viral pathogen and a significantly higher rate of co-infections with multiple viruses, when compared to all other patients >5 years of age. Influenza B, adenovirus, and respiratory syncytial virus were all detected at significantly higher rates in children <5 years of age. This study confirmed that multiple respiratory viruses are circulating and contributing to the presentation of influenza-like-illness in Papua New Guinea. PMID:24136362

Kono, Jacinta; Jonduo, Marinjho H; Omena, Matthew; Siba, Peter M; Horwood, Paul F

2014-05-01

95

Identification of a novel HLA-A*24:02-restricted adenovirus serotype 11-specific CD8+ T-cell epitope for adoptive immunotherapy.  

PubMed

Subgroup B adenovirus serotype 11 (Ad11) occasionally causes fatal infections in immunocompromised patients. The present study describes a novel Ad11 epitope presented by HLA-A*24:02 that could be used for adoptive immunotherapy. Ten synthetic Ad11 hexon protein-derived nonamer peptides that bound to HLA-A*24:02 were selected by a computer algorithm and MHC stabilization assay. Stimulation of peripheral blood mononuclear cells from HLA-A*24:02+ donors with each of these synthetic peptides induced peptide-specific CD8(+) T-cells for three peptides. Testing the reactivity of these peptide-specific CD8(+) T-cells against various target cells confirmed that peptide TYFNLGNKF is naturally processed in Ad11-infected cells and is presented by HLA-A*24:02. Emergence of TYFNLGNKF-specific CD8(+) T-cells coincided with the clearance of adenoviruses in a patient with Ad11 disease. Importantly, TYFNLGNKF-specific CD8(+) T-cells were suggested to be not serotype cross-reactive. The novel HLA-A*24:02-restricted Ad11 epitope could be used for anti-Ad11 adoptive immunotherapy and to monitor immunity to Ad11 using MHC tetramers. PMID:23911395

Imahashi, Nobuhiko; Nishida, Tetsuya; Ito, Yoshinori; Kawada, Jun-ichi; Nakazawa, Yozo; Toji, Shingo; Suzuki, Susumu; Terakura, Seitaro; Kato, Tomonori; Murata, Makoto; Naoe, Tomoki

2013-12-01

96

Evaluation of the Temporal Association between Kawasaki Disease and Viral Infections in South Korea  

PubMed Central

Background and Objectives This study is aimed at elucidating potential temporal associations between the occurrence of Kawasaki disease (KD) and various viral infections. Subjects and Methods We obtained monthly patterns of KD from the seventh nationwide survey and viral detection data from the Korea Centers for Disease Control and Prevention from 2009 to 2011 and evaluated temporal correlations between them for each month. The respiratory viruses detected using a multiplex real-time-polymerase chain reaction kit were influenza virus (A/H1N1, A/H3N2, A/H5N1, and B), adenovirus, parainfluenza virus (type 1, 2, 3), respiratory syncytial virus (type A, B), human rhinovirus, human coronavirus (OC43/229E, NL63), human bocavirus, and enterovirus. Results We obtained data from a total of 13031 patients who were treated for acute KD from 87 hospitals with pediatric residence programs. During this survey, KD showed highest overall incidence in summer and winter seasons and lowest incidence in February and October. We received viral detection data for a total of 14267 patients. Viral detection was highest during winter and spring seasons. The most commonly detected virus was human rhinovirus (32.6%), followed by influenza virus (26.8%). The monthly incidence of KD showed significant correlation with the monthly overall viral detection (p=0.022, r=0.382). In particular, human bocavirus and enterovirus have significant correlations with monthly patterns of KD occurrence (p=0.032 and p=0.007, respectively) and influenza virus correlated with KD occurrence with borderline significance (p=0.063). Conclusion The temporal association between monthly occurrence of KD and viral detection suggests the etiologic importance of precedent infection in the development of KD.

Kim, Gi Beom; Park, Sohee; Han, Ji Whan; Park, Yong Won; Hong, Young Mi

2014-01-01

97

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: Jennifer.grosjean@imperial.ac.uk; 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)

2006-02-17

98

ONYX-411, a conditionally replicative oncolytic adenovirus, induces cell death in anaplastic thyroid carcinoma cell lines and suppresses the growth of xenograft tumors in nude mice.  

PubMed

Anaplastic thyroid carcinoma (ATC) is the most aggressive thyroid cancer variant, accounting for 1-2% of all cases, but 33% of deaths, and exhibiting an average life expectancy of 5 months. ATC is largely unresponsive to radioactive iodine, chemotherapy, external beam radiation or surgery, underscoring the need for new and effective therapies. We evaluated the therapeutic potential of an oncolytic adenovirus, ONYX-411, that replicates selectively in and kills cells with dysfunction of the retinoblastoma (RB) pathway. In the present study, we report that ONYX-411 is able to induce cell death in eight human anaplastic carcinoma cell lines in vitro. The cytopathic effect of the virus is specific to cells with RB dysfunction, which appears to be frequent in ATC. We confirmed the expression of the coxsackie adenovirus receptor, CAR, in all ATC cell lines, demonstrating the potentially universal application of this oncolytic viral therapy to ATC. In addition, the growth of xenograft tumors induced in athymic mice with the ARO and DRO cell lines was significantly reduced by ONYX-411 treatment. These results indicate that ONYX-411 can be a potential therapeutic agent for the treatment of ATC, rendering this class of conditionally replicating adenoviruses an attractive candidate for clinical trials. PMID:18583996

Reddi, H V; Madde, P; Reichert-Eberhardt, A J; Galanis, E C; Copland, J A; McIver, B; Grebe, S K G; Eberhardt, N L

2008-11-01

99

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

PubMed

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

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

2013-12-01

100

Sesquiterpenes and alkaloids from the roots of Alangium chinense.  

PubMed

Four new sesquiterpenes (1-4), four new alkaloids (5a, 6a, 6b, and 7), and nine known compounds (5b and 8-15) were isolated from an ethanolic extract of roots of Alangium chinense. The structure of 1 was confirmed by X-ray crystallography. The configurations of 5 and 6 were assigned by chiral HPLC analysis and CD spectra. Compounds 3, 4, 8-13, and 15 exhibited antiviral activity against Coxsackie virus B3 with IC50 values of 1.4-15.4 ?M. Compounds 2-4, 7, and 9-13 showed antioxidant activities against Fe(2+)-cysteine-induced rat liver microsomal lipid peroxidation, with IC50 values of 3.8-45.7 ?M. Compound 5b displayed neuritis inhibitory activity against microglial inflammation factor, with an IC50 value of 6.7 ?M. None of the compounds exhibited detectable cytotoxic activity toward any of five tumor cell lines (A549, Be-17402, BGC-823, HCT-8, and A2780) in the MTT assay. PMID:23734721

Zhang, Yan; Liu, Yun-Bao; Li, Yong; Ma, Shuang-Gang; Li, Li; Qu, Jing; Zhang, Dan; Chen, Xiao-Guang; Jiang, Jian-Dong; Yu, Shi-Shan

2013-06-28

101

Adenovirus gene transfer to amelogenesis imperfecta ameloblast-like cells.  

PubMed

To explore gene therapy strategies for amelogenesis imperfecta (AI), a human ameloblast-like cell population was established from third molars of an AI-affected patient. These cells were characterized by expression of cytokeratin 14, major enamel proteins and alkaline phosphatase staining. Suboptimal transduction of the ameloblast-like cells by an adenovirus type 5 (Ad5) vector was consistent with lower levels of the coxsackie-and-adenovirus receptor (CAR) on those cells relative to CAR-positive A549 cells. To overcome CAR -deficiency, we evaluated capsid-modified Ad5 vectors with various genetic capsid modifications including "pK7" and/or "RGD" motif-containing short peptides incorporated in the capsid protein fiber as well as fiber chimera with the Ad serotype 3 (Ad3) fiber "knob" domain. All fiber modifications provided an augmented transduction of AI-ameloblasts, revealed following vector dose normalization in A549 cells with a superior effect (up to 404-fold) of pK7/RGD double modification. This robust infectivity enhancement occurred through vector binding to both ?(v)?3/?(v)?5 integrins and heparan sulfate proteoglycans (HSPGs) highly expressed by AI-ameloblasts as revealed by gene transfer blocking experiments. This work thus not only pioneers establishment of human AI ameloblast-like cell population as a model for in vitro studies but also reveals an optimal infectivity-enhancement strategy for a potential Ad5 vector-mediated gene therapy for AI. PMID:22003382

Borovjagin, Anton V; Dong, Juan; Passineau, Michael J; Ren, Changchun; Lamani, Ejvis; Mamaeva, Olga A; Wu, Hongju; Keyser, Enid; Murakami, Miho; Chen, Shuo; MacDougall, Mary

2011-01-01

102

Pseudotyping the adenovirus serotype 5 capsid with both the fibre and penton of serotype 35 enhances vascular smooth muscle cell transduction.  

PubMed

Ex vivo gene therapy during coronary artery bypass grafting (CABG) holds great potential to prevent excessive smooth muscle cell (SMC) proliferation, neointima formation and graft failure. The most successful preclinical strategies to date have utilised vectors based on the species C adenovirus, Ad5, which engages the Coxsackie and Adenovirus receptor (CAR) as its primary attachment receptor. Profiling receptors on human SMCs demonstrated the absence of CAR but substantial expression of the species B receptor CD46. We performed transduction experiments using Ad5 and the CD46-utilising adenovirus Ad35, and found Ad35 significantly more efficient at transducing SMCs. To evaluate whether transduction could be further augmented, we evaluated chimeric CD46-utilising Ad5/Ad35 vectors comprising the Ad5 capsid pseudotyped with the Ad35 fibre alone (Ad5/F35) or in combination with the Ad35 penton (Ad5/F35/P35). In human smooth muscle cells (hSMCs), Ad5/F35/P35 mediated significantly higher levels of transduction than either parental vector or Ad5/F35. Ex vivo transduction experiments using mouse aortas from CD46 transgenics demonstrated that Ad5/F35/P35 was significantly more efficient at transducing SMCs than the other vectors tested. Finally, ex vivo transduction and immunofluorescent colocalisation experiments using human tissue from CABG procedures confirmed the preclinical potential of Ad5/F35/P35 as an efficient vector for vascular transduction during CABG. PMID:24005577

Parker, A L; White, K M; Lavery, C A; Custers, J; Waddington, S N; Baker, A H

2013-12-01

103

Isolation of enteroviruses from water, suspended solids, and sediments from Galveston Bay: survival of poliovirus and rotavirus adsorbed to sediments.  

PubMed Central

The distribution and quantitation of enteroviruses among water, suspended solids, and compact sediments in a polluted estuary are described. Samples were collected sequentially from water, suspended solids, fluffy sediments (uppermost layer of bottom sediments), and compact sediment. A total of 103 samples were examined of which 27 (26%) were positive for virus. Polioviruses were recovered most often, followed by coxsackie B viruses and echoviruses 7 and 29. Virus was found most often attached to suspended solids: 72% of these samples were positive, whereas only 14% of water samples without solids yielded virus. Fluffy sediments yielded virus in 47% of the samples, whereas only 5% of compact bottom-sediment samples were positive. When associated with solids, poliovirus and rotavirus retained their infectious quality for 19 days. The same viruses remained infectious for only 9 days when freely suspended in seawater. Collection of suspended solids at ambient water pH appears to be very useful for the detection of virus; it has advantages over collecting and processing large volumes of water, with accompanying pH adjustment and salt addition for processing.

Rao, V C; Seidel, K M; Goyal, S M; Metcalf, T G; Melnick, J L

1984-01-01

104

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

PubMed

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

2013-07-19

105

Efficient targeting of adenoviral vectors to integrin positive vascular cells utilizing a CAR-cyclic RGD linker protein.  

PubMed

Vascular smooth muscle (VSMC) and endothelial cells (EC) are particularly resistant to infection by type 5 adenovirus (Ad) vectors. To overcome this limitation and target Ad vectors to ubiquitously expressed alpha(V)beta(3/5) integrins, we have generated a linker protein consisting of the extracellular domain of the coxsackie adenovirus receptor (CAR) connected via avidin to a biotinylated cyclic (c) RGD peptide. After optimization of CAR to cRGD and to Ad coupling, infection of mouse heart endothelial cells (H5V) could be augmented significantly, as demonstrated by 600-fold increased transgene expression levels. In EOMAs, a hemangioendothelioma-derived cell line, the fraction of infected cells was enhanced 4- to 6-fold. Furthermore, the fraction of infected primary mouse VSMC was increased from virtually 0% to 25%. Finally, in human umbilical vein endothelial cells, the number of GFP positive cells was enhanced from 2% to 75%. In conclusion, CAR-cRGD is a versatile and highly efficient construct to target Ad vectors to both transformed and primary VSMC and EC. PMID:16259946

Krom, Y D; Gras, J C E; Frants, R R; Havekes, L M; van Berkel, T J; Biessen, E A L; van Dijk, K Willems

2005-12-16

106

Therapeutic efficacy of an oncolytic adenovirus containing RGD ligand in minor capsid protein IX and Fiber, ?24DoubleRGD, in an ovarian cancer model  

PubMed Central

Ovarian cancer is the leading cause of gynecological disease death despite advances in medicine. Therefore, novel strategies are required for ovarian cancer therapy. Conditionally replicative adenoviruses (CRAds), genetically modified as anti-cancer therapeutics, are one of the most attractive candidate agents for cancer therapy. However, a paucity of coxsackie B virus and adenovirus receptor (CAR) expression on the surface of ovarian cancer cells has impeded treatment of ovarian cancer using this approach. This study sought to engineer a CRAd with enhanced oncolytic ability in ovarian cancer cells, “?24DoubleRGD.” ?24DoubleRGD carries an arginine-glycine-aspartate (RGD) motif incorporated into both fiber and capsid protein IX (pIX) and its oncolytic efficacy was evaluated in ovarian cancer. In vitro analysis of cell viability showed that infection of ovarian cancer cells with ?24DoubleRGD leads to increased cell killing relative to the control CRAds. Data from this study suggested that not only an increase in number of RGD motifs on the CRAd capsid, but also a change in the repertoir of targeted integrins could lead to enhanced oncolytic potency of ?24DoubleRGD in ovarian cancer cells in vitro. In an intraperitoneal model of ovarian cancer, mice injected with ?24DoubleRGD showed, however, a similar survival rate as mice treated with control CRAds.

Gamble, Lena J.; Ugai, Hideyo; Wang, Minghui; Borovjagin, Anton V.; Matthews, Qiana L.

2013-01-01

107

[The inhibitory effects of catechin derivatives on the activities of human immunodeficiency virus reverse transcriptase and DNA polymerases].  

PubMed

Catechin derivatives including (-)-epicatechin gallate (ECG), (-)-epigallocatechin gallate (EGCG), (-)-epigallocatechin (EGC) and green tea extract (GTE) were found to inhibit the activities of cloned human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT), duck hepatitis B virus replication complexes reverse transcriptase (DHBV RCs RT), herpes simplex virus 1 DNA polymerase (HSV-1 DNAP) and cow thymus DNA polymerase alpha (CT DNAP alpha). EGCG and ECG were shown to be very potent inhibitors of HIV-1 RT. According to the IC50 values for HIV-1 RT, these compounds can be ordered as EGCG 0.0066 mumol/L > ECG 0.084 mumol/L > GTE 0.1 microgram/ml > EGC 7.2 mumol/L. DHBV RCs RT was the least sensitive to these compounds. Kinetic study showed that EGCG exerts a mixed inhibition with respect to external template inducer poly (rA).oligo (dT) 12-18 and a noncompetitive inhibition with respect to substrate dTTP for HIV-1 RT. Bovine serum albumin significantly reduced the inhibitory effects of catechin analogues and GTE on HIV-1 RT. In tissue culture GTE inhibited the cytopathic effect of coxsackie B3 virus, but did not inhibit the cytopathic effects of HSV-1, HSV-2, influenza A or influenza B viruses. PMID:1284389

Tao, P

1992-10-01

108

Screening of some Tanzanian medicinal plants from Bunda district for antibacterial, antifungal and antiviral activities.  

PubMed

Extracts from 50 plant parts obtained from 39 different plants belonging to 22 families used to treat infectious diseases in Bunda district, Tanzania, were screened against twelve microorganisms, including the bacteria Bacillus cereus, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae and Salmonella typhimurium, the fungi Aspergillus niger, Candida albicans, and the viruses Herpes Simplex Virus type 1, Vesicular Stomatitis Virus T2, Coxsackie B2 and Semliki Forest A7. The highest activity was obtained for the n-hexane extract of Elaeodendron schlechteranum root bark against the Gram-positive bacteria Bacillus cereus (MIC 0.97 microg/ml and MBC 1.95 microg/ml) and Staphylococcus aureus (MIC 3.90 microg/ml and MBC 31.25 microg/ml). Gram-negative bacteria were less sensitive. Only Balanites aegyptiaca stem bark exhibited a high antifungal activity against Candida albicans (MIC 125 microg/ml and MFC 250 microg/ml). Extracts from four plants; Lannea schweinfurthii, Combretum adenogonium, Ficus sycomorus and Terminalia mollis showed strong antiviral activity with RF values of 10(3) and 10(4) against Herpes Simplex Virus type 1 at various concentrations. Our results support, at least in part, the use of most plants as claimed by traditional healers/informants especially against the Gram-positive bacteria Bacillus cereus and Staphylococcus aureus. PMID:18582554

Maregesi, Sheila Mgole; Pieters, Luc; Ngassapa, Olipa David; Apers, Sandra; Vingerhoets, Rita; Cos, Paul; Berghe, Dirk A Vanden; Vlietinck, Arnold J

2008-09-01

109

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

PubMed

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

2014-01-30

110

ONYX-411, a conditionally replicative oncolytic adenovirus, induces cell death in anaplastic thyroid carcinoma cell lines and suppresses the growth of xenograft tumors in nude mice  

PubMed Central

Anaplastic thyroid carcinoma (ATC) is the most aggressive thyroid cancer variant, accounting for 1–2% of all cases, but 33% of deaths, and exhibiting an average life expectancy of 5 months. ATC is largely unresponsive to radioactive iodine, chemotherapy, external beam radiation or surgery, underscoring the need for new and effective therapies. We evaluated the therapeutic potential of an oncolytic adenovirus, ONYX-411, that replicates selectively in and kills cells with dysfunction of the retinoblastoma (RB) pathway. In the present study, we report that ONYX-411 is able to induce cell death in eight human anaplastic carcinoma cell lines in vitro. The cytopathic effect of the virus is specific to cells with RB dysfunction, which appears to be frequent in ATC. We confirmed the expression of the coxsackie adenovirus receptor, CAR, in all ATC cell lines, demonstrating the potentially universal application of this oncolytic viral therapy to ATC. In addition, the growth of xenograft tumors induced in athymic mice with the ARO and DRO cell lines was significantly reduced by ONYX-411 treatment. These results indicate that ONYX-411 can be a potential therapeutic agent for the treatment of ATC, rendering this class of conditionally replicating adenoviruses an attractive candidate for clinical trials.

Reddi, HV; Madde, P; Reichert-Eberhardt, AJ; Galanis, EC; Copland, JA; McIver, B; Grebe, SKG; Eberhardt, NL

2011-01-01

111

Model-driven Approaches for in Vitro Combination Therapy using ONYX-015 Replicating Oncolytic Adenovirus 1  

PubMed Central

Replicating genetically modified adenoviruses have shown promise as a new treatment approach against cancer. Recombinant adenoviruses replicate only in cancer cells which contain certain mutations, such as the loss of functional p53, as is the case in the virus ONYX-015. The successful entry of the viral particle into target cells is strongly dependent on the presence of the main receptor for adenovirus, the coxsackie- and adeno-virus receptor (CAR). This receptor is frequently down-regulated in highly malignant cells, rendering this population less vulnerable to viral attack. It has been shown that use of MEK inhibitors can up-regulate CAR expression, resulting in enhanced adenovirus entry into the cells. However, inhibition of MEK results in G1 cell cycle arrest, rendering infected cells temporarily unable to produce virus. This forces a tradeo1. While drug mediated up-regulation of CAR enhances virus entry into cancer cells, the consequent cell cycle arrest inhibits production of new virus particles and the replication of the virus. Optimal control-based schedules of MEK inhibitor application should increase the efficacy of this treatment, maximizing the overall tumor toxicity by exploiting the dynamics of CAR expression and viral production. We introduce a mathematical model of these dynamics and show simple optimal control based strategies which motivate this approach.

Zurakowski, Ryan; Wodarz, Dominik

2009-01-01

112

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

PubMed Central

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

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

2014-01-01

113

Fiber-mutant technique can augment gene transduction efficacy and anti-tumor effects against established murine melanoma by cytokine-gene therapy using adenovirus vectors.  

PubMed

Melanoma cells are relatively resistant to adenovirus vector (Ad)-mediated gene transfer due to the low expression of Coxsackie-adenovirus receptor (CAR), which acts as a primitive Ad-receptor. Therefore, extremely high doses of Ad are required for effective gene therapy against melanoma. In the present study, we investigated whether fiber-mutant Ad containing the Arg-Gly-Asp (RGD) sequence in the fiber knob could promote gene delivery and anti-tumor effects in the murine B16 BL6 tumor model. B16 BL6 cells (in vitro) and tumors (in vivo) infected with RGD fiber-mutant Ad containing a tumor necrosis factor alpha gene (Ad-RGD-TNFalpha) produced more TNFalpha than those infected with conventional Ad-TNFalpha. In addition, Ad-RGD-TNFalpha required about one-tenth the dosage of Ad-TNFalpha for induction of equal therapeutic effects upon intratumoral injection into established B16 BL6 tumors. Furthermore, the combination of both TNFalpha- and interleukin 12-expressing RGD fiber-mutant Ads exhibited more effective tumor regression than the Ad expressing each alone. These results suggested that the fiber-mutant for altering Ad-tropism is a very potent technology for advancing gene therapy for melanoma. PMID:11809531

Okada, Yuka; Okada, Naoki; Nakagawa, Shinsaku; Mizuguchi, Hiroyuki; Kanehira, Makiko; Nishino, Naoko; Takahashi, Koichi; Mizuno, Nobuyasu; Hayakawa, Takao; Mayumi, Tadanori

2002-03-01

114

Tumor necrosis factor alpha-gene therapy for an established murine melanoma using RGD (Arg-Gly-Asp) fiber-mutant adenovirus vectors.  

PubMed

Although adenovirus vectors (Ad) provide high-level transduction efficacy to many cell types, extremely high doses of Ad are required for sufficient gene transduction into several tumors, including melanoma. Here, we demonstrated that the expression of coxsackie-adenovirus receptor, a primitive Ad-receptor, was very low in murine and human melanoma cells. We also found that fiber-mutant Ad containing the Arg-Gly-Asp (RGD) sequence in the fiber knob remarkably augmented gene transduction efficacy in melanoma cells by targeting alpha(v)-integrins. In addition, intratumoral injection of RGD fiber-mutant Ad containing the tumor necrosis factor alpha gene (Ad-RGD-TNFalpha) revealed dramatic anti-tumor efficacy through hemolytic necrosis in an established murine B16 BL6 melanoma model. Ad-RGD-TNFalpha required one-tenth the dosage of Ad-TNFalpha to induce an equal therapeutic effect. These results suggest that alpha(v)-integrin-targeted Ad will be a very powerful tool for the advancement of melanoma gene therapy. PMID:11985794

Okada, Yuka; Okada, Naoki; Nakagawa, Shinsaku; Mizuguchi, Hiroyuki; Takahashi, Koichi; Mizuno, Nobuyasu; Fujita, Takuya; Yamamoto, Akira; Hayakawa, Takao; Mayumi, Tadanori

2002-04-01

115

[Epidemiologic characteristics of human parvovirus B19. Report of an epidemic of erythema infectiosum in the area of Belgrade].  

PubMed

During the period 29.10.1987-10.5.1988 an outbreak of an illness with rash resembling Erythema Infectiosum occurred among school children in one part of Belgrade-Mladenovac. This is the first outbreak of Erythema Infectiosum reported in Yugoslavia and confirmed in University College of Middlesex School of Medicine, London, in November 1989. Of 720 school children exposed to infection of Human Parvovirus B 19 two hundred and eighty four (39.4%) have had clinical symptoms and 166 (58.4%) of them were in the age group 10-14. The sero-epidemiologic investigations excluded infection of Rubella virus, Cytomegalovirus and Coxsackie viruses A 9 and B 5. To detect the infection Elisa IgM antibody, complement fixation, inhibition of hemagglutination and isolation have been used. Clinical symptoms of illness have been manifested as Rubella like exanthemas on extremities in 92.5%, extremities and body in 26.5% and the phenomenon "slapped Cheek" was discovered in 76.25% of all patients. Reappearance of rash has been observed in 25% of cases. PMID:1465672

Bukumirovi?, K; Radosavljevi?, M; Milki?, V; Antonijevi?, B; Polak, D; Dodi?, M; Kanjuh, B

1992-01-01

116

Isolation, identification and antiviral activities of metabolites of calycosin-7-O-?-D-glucopyranoside.  

PubMed

In vivo and in vitro metabolites of calycosin-7-O-?-D-glucopyranoside in rats were identified using a specific and sensitive high performance liquid chromatography-tandem mass spectrometry (HPLC-MS(n)) method. The parent compound and twelve metabolites were found in rat urine after oral administration of calycosin-7-O-?-D-glucopyranoside. The parent compound and six metabolites were detected in rat plasma. In heart, liver, spleen, lung and kidney samples, respectively, six, eight, seven, nine and nine metabolites were identified, in addition to the parent compound. Three metabolites, but no trace of parent drug, were found in the rat intestinal flora incubation mixture and feces, which demonstrated cleavage of the glycosidic bond of the parent compound in intestines. The main phase I metabolic pathways of calycosin-7-O-?-D-glucopyranoside in rats were deglycosylation, dehydroxylation and demethylation reactions; phase II metabolism included sulfation, methylation, glucuronidation and glycosylation (probably). Furthermore, two metabolites commonly found in rat urine, plasma and tissues were isolated from feces and characterized by NMR. The antiviral activities of the metabolite calycosin against coxsackie virus B? (CVB?) and human immunodeficiency virus (HIV) were remarkably stronger than those of calycosin-7-O-?-D-glucopyranoside. PMID:21703796

Chen, Luying; Li, Zhixiong; Tang, Yihong; Cui, Xiaolan; Luo, Ronghua; Guo, Shanshan; Zheng, Yongtang; Huang, Chenggang

2011-09-10

117

T-lymphocyte mediated injury of beta cells in dual-aetiology diabetes mellitus in Swiss albino mouse.  

PubMed

Earlier we had described a dual aetiology diabetes mellitus (DADM) in mice injected with a sub-diabetogenic dose of streptozotocin (SD-SZN) and afterwards infected with coxsackie B3 virus (CBV). Further experiments were conducted to understand the mechanism of diabetogenesis. In in vitro stimulation and proliferation tests, the splenic lymphocytes (SLC) of mice given either SD-SZN or CBV infection showed lower responses to two T cell mitogens than those of control mice, indicating an immunosuppressive effect. Unexpectedly, SLC of mice given both SD-SZN and CBV showed enhanced response, indicating immunoactivation; they were not stimulated to proliferation in response to CBV antigen, indicating that the immunoactivation was not directed against CBV, but against streptozotocin or cellular elements. When mice were depleted of T cells by injecting with anti-thymocyte serum, the diabetogenic effect of SD-SZN and CBV infection was abrogated, without diminishing the replication of virus in the pancreas. Thus beta cell injury in DADM appears to be T cell-mediated. PMID:1335964

Babu, P G; Saraswathi, N K; John, T J

1992-09-01

118

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

PubMed Central

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

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

2014-01-01

119

Intralesional immunotherapy for melanoma.  

PubMed

Intralesional immunotherapy of melanoma has two complementary aims. One is to cause regression of the injected metastasis. The other is to incite or modulate systemic immune responses in such a way that non-injected metastases will also undergo regression. A number of phase 1 and phase II studies with cytokines, viral, or bacterial agents have been conducted but their use has remained sporadic and has not progressed to become established treatments. Two treatments have progressed to randomized phase III studies. The most promising of these is based on intralesional injection of a genetically modified herpes simplex virus (HSV) (T-Vec). Initial results have shown a significant effect on durable response rates (DRR) but effects on overall survival remain under study. The second involved injection of plasmids coding for the HLA B7 antigen (Allovectin). Despite encouraging early results the treatment did not reach its endpoints and its use has been discontinued. A phase II study involving intralesional injection of oncolytic A21 coxsackie virus (Cavatak) is also under way and is showing promise. PMID:24301265

Hersey, Peter; Gallagher, Stuart

2014-03-01

120

The action of three antiseptics/disinfectants against enveloped and non-enveloped viruses.  

PubMed

The antiviral action of chloroxylenol, benzalkonium chloride and cetrimide/chlorhexidine was assessed against a range of enveloped and non-enveloped human viruses using a suspension test method. Viral suspensions of 10(6)-10(7) pfu/TCID50 or sfu were prepared in each of the antiseptic/disinfectant solutions in the presence of a bovine serum/yeast extract mixture to simulate 'dirty conditions'. During incubation, aliquots were removed at predetermined timepoints up to 10 min to assess the kinetics of inactivation. Results indicate that all products were effective in inactivating the enveloped viruses herpes simplex virus type 1 and human immunodeficiency virus type 1, whilst being ineffective in inactivating human coronavirus, also enveloped, and the non-enveloped viruses. The exception to this was the benzalkonium chloride-based product (Dettol Hospital Concentrate) which was active against the non-enveloped human coxsackie virus. Four antiseptic/disinfectant solutions with chloroxylenol, benzalkonium chloride, cetrimide/chlorhexidine and povidone-iodine were also assessed for antiviral effect against human immunodeficiency virus in the presence of whole human blood. All four solutions proved to be effective within 1 min despite the cytotoxic nature of the compounds to the detection system. PMID:9602977

Wood, A; Payne, D

1998-04-01

121

Improving gene transfer in human renal carcinoma cells: Utilization of adenovirus vectors containing chimeric type 5 and type 35 fiber proteins  

PubMed Central

The transduction efficacy of adenovirus serotype 5 (Ad5) vector in human renal carcinoma cells is generally low due to the down-regulated expression of Coxsackie and adenovirus receptor (CAR) in target cells. By contrast, the infectivity of adenovirus serotype 35 vectors depends on the binding rate to CD46 receptor, independent of CAR. In this study, we examined whether an adenovirus vector containing chimeric type 5 and type 35 fiber proteins (Ad5/F35) increases transduction efficiency compared to Ad5 vector in human renal carcinoma cells in vitro. The expression of CAR was much lower in the human renal carcinoma cells than in control HEK293 cells. By contrast, the expression of CD46 was similar and perhaps at a higher level in the human renal carcinoma cells than in the HEK293 cells. The transduction efficacy of Ad5/F35 vector was dramatically higher compared to that of Ad5 in human renal carcinoma cells, and was correlated to the expression of CD46. Thus, Ad5/35 vector may be useful for the development of novel gene therapy approaches to renal cell carcinoma.

ACHARYA, BISHNU; TERAO, SHUJI; SUZUKI, TORU; NAOE, MICHIO; HAMADA, KATSUYUKI; MIZUGUCHI, HIROYUKI; GOTOH, AKINOBU

2010-01-01

122

Borna disease virus nucleoprotein inhibits type I interferon induction through the interferon regulatory factor 7 pathway  

SciTech Connect

Highlights: •IRF7 nuclear localisation was inhibited by BDV persistently infected. •BDV N protein resistant to IFN induction both in BDV infected OL cell and N protein plasmid transfected OL cell. •BDV N protein is related to the inhibition of IRF7 nuclear localisation. -- Abstract: The expression of type I interferon (IFN) is one of the most potent innate defences against viral infection in higher vertebrates. Borna disease virus (BDV) establishes persistent, noncytolytic infections in animals and in cultured cells. Early studies have shown that the BDV phosphoprotein can inhibit the activation of type I IFN through the TBK1–IRF3 pathway. The function of the BDV nucleoprotein in the inhibition of IFN activity is not yet clear. In this study, we demonstrated IRF7 activation and increased IFN-?/? expression in a BDV-persistently infected human oligodendroglia cell line following RNA interference-mediated BDV nucleoprotein silencing. Furthermore, we showed that BDV nucleoprotein prevented the nuclear localisation of IRF7 and inhibited endogenous IFN induction by poly(I:C), coxsackie virus B3 and IFN-?. Our findings provide evidence for a previously undescribed mechanism by which the BDV nucleoprotein inhibits type I IFN expression by interfering with the IRF7 pathway.

Song, Wuqi [The Heilongjiang Key Laboratory of Immunity and Infection, Heilongjiang (China) [The Heilongjiang Key Laboratory of Immunity and Infection, Heilongjiang (China); Department of Microbiology, Harbin Medical University (China); Kao, Wenping [The Key Laboratory of Pathogenic Biology, Heilongjiang Higher Education Institutions (China) [The Key Laboratory of Pathogenic Biology, Heilongjiang Higher Education Institutions (China); Department of Microbiology, Harbin Medical University (China); Zhai, Aixia [The Heilongjiang Key Laboratory of Immunity and Infection, Heilongjiang (China)] [The Heilongjiang Key Laboratory of Immunity and Infection, Heilongjiang (China); Qian, Jun; Li, Yujun [The Key Laboratory of Pathogenic Biology, Heilongjiang Higher Education Institutions (China)] [The Key Laboratory of Pathogenic Biology, Heilongjiang Higher Education Institutions (China); Zhang, Qingmeng [The Heilongjiang Key Laboratory of Immunity and Infection, Heilongjiang (China)] [The Heilongjiang Key Laboratory of Immunity and Infection, Heilongjiang (China); Zhao, Hong; Hu, Yunlong; Li, Hui [Department of Microbiology, Harbin Medical University (China)] [Department of Microbiology, Harbin Medical University (China); Zhang, Fengmin, E-mail: fengminzhang@ems.hrbmu.edu.cn [The Heilongjiang Key Laboratory of Immunity and Infection, Heilongjiang (China) [The Heilongjiang Key Laboratory of Immunity and Infection, Heilongjiang (China); The Key Laboratory of Pathogenic Biology, Heilongjiang Higher Education Institutions (China); Department of Microbiology, Harbin Medical University (China)

2013-09-06

123

[A study on enterovirus infection in Osaka prefecture --comparison with virus isolation and RT-PCR amplifying viral protein 1 region].  

PubMed

Detection of viral genomes with RT-PCR, which amplifies viral protein 1 region, as well as virus isolation was performed on 860 patients (996 specimens) who had been suspected for enterovirus (EV) infection in Osaka Prefecture from April 2003 to Jury 2004. The viral positive rates of the clinical materials, combining above two procedures, were as follows: 48.2% from the feces, 38.3% from the throat swabs, and 18.0% from the cerebrospinal fluids. The positive rate by the clinical diagnosis indicated that herpangina was the highest at 44.7%, while encephalitis was the lowest at 13.4%. Out of the all specimens, the viral positive rate of RT-PCR varied from 22.3 to 24.7%, while that of virus isolation was 15.6%. The total viral positive rate of both procedures combined was 29.8%. Since the detection of EV by RT-PCR amplifing viral protein 1 region was more rapid than virus isolation and superior in detection of coxsackie group A virus, RT-PCR is an effective procedure for diagnosis of herpangina. PMID:15977548

Yamazaki, Kenji; Otake, Toru

2005-02-01

124

Enhanced Gene Delivery to Human Primary Endothelial Cells Using Tropism-Modified Adenovirus Vectors.  

PubMed

Endothelial cells have been noted to have relatively low expression of the native receptor for adenovirus serotype 5 (Ad5), coxsackie and adenovirus receptor (CAR), and are thus refractory to Ad5 infection. In this study, we hypothesize that increases in the infectivity of Ad5 in primary human pulmonary artery (HPAEC), coronary artery (HCAEC) and umbilical vein endothelial cells (HUVEC) can be achieved through genetic capsid modification of Ad5 to bypass CAR-dependent infection. The modifications tested in this study include incorporation of an integrin-binding RGD peptide motif (Ad5.RGD), a poly-lysine motif (Ad5.pK7), a combination of both of these peptide domains (Ad5.RGD.pK7), an adenovirus serotype 3 knob domain (Ad5/3Luc1) and canine adenovirus serotype 1 or 2 knob domains (Ad5Luc1-CK1 and Ad5Luc1-CK2). In HPAEC and HCAEC, the greatest infectivity enhancements were achieved using Ad5/3Luc1 (26-fold and 30-fold respectively). HUVEC was most readily infected by Ad5Luc1-CK1 (213-fold). These results demonstrate that gains in Ad5 infectivity in endothelial cells can be accomplished with genetic capsid modifications. PMID:19834585

Preuss, Meredith A; Glasgow, Joel N; Everts, Maaike; Stoff-Khalili, Mariam A; Wu, Hongju; Curiel, David T

2008-01-01

125

Infection and stillbirth  

PubMed Central

Summary Infection may cause stillbirth by several mechanisms, including direct infection, placental damage, and severe maternal illness. Various organisms have been associated with stillbirth, including many bacteria, viruses, and protozoa. In developed countries, between 10% and 25% of stillbirths may be caused by an infection, whereas in developing countries, which have much higher stillbirth rates, the contribution of infection is much greater. In developed countries, ascending bacterial infection, both before and after membrane rupture, with organisms such as Escherichia coli, group B streptococci, and Ureaplasma urealyticum is usually the most common infectious cause of stillbirth. However, in areas where syphilis is prevalent, up to half of all stillbirths may be caused by this infection alone. Malaria may be an important cause of stillbirth in women infected for the first time in pregnancy. The two most important viral causes of stillbirth are parvovirus and Coxsackie virus, although a number of other viral infections appear to be causal. Toxoplasma gondii, leptospirosis, Listeria monocytogenes, and the organisms that cause leptospirosis, Q fever, and Lyme disease have all been implicated as etiologic for stillbirth. In certain developing countries, the stillbirth rate is high and the infection-related component so great that achieving a substantial reduction in stillbirth should be possible by reducing maternal infections. However, because infection-related stillbirth is uncommon in developed countries, and because those that do occur are caused by a wide variety of organisms, reducing this etiologic component of stillbirth much further will be difficult.

McClure, Elizabeth M.; Goldenberg, Robert L.

2014-01-01

126

Echoviruses include genetically distinct serotypes.  

PubMed

We have studied the genetic relationships of echoviruses using nucleotide sequencing and hybridization analysis. The nucleotide sequence of the echovirus 11 (EV11) P2 and P3 regions, which encode the nonstructural proteins, was shown to resemble closely those of coxsackie B viruses (CBV) and coxsackievirus A9 (CAV9). EV11, CBV and CAV9 have a similar organization in the 3' non-coding region when compared to polioviruses and CAV21. In contrast, the 3' end of EV22 shares only minimal sequence homology with other sequenced enteroviruses, and the 3' non-coding region has a unique secondary structure. Thirty-three echovirus reference strains were tested by nucleic acid hybridization using cDNA probes from the genomes of EV6, 11, 18 and 22. It was shown that a great majority of the strains belongs to the same subgroup as serotypes 6, 11 and 18, whereas EV22 and EV23 are genetically not closely related to this major subgroup. PMID:2170575

Auvinen, P; Hyypiä, T

1990-09-01

127

Stabilization of isometric DNA viruses against thermoinactivation by lowered ionic strength.  

PubMed

The thermostability of isometric DNA viruses increases, if the ionic strength is diminished before heating. When unpurified virus material from cell cultures is heated under conditions (temperature, time) which lead to a reduction in infectivity by 3 to 4 log10, this loss in titer is 1 to 3 log10 less, if the ionic strength is decreased by diluting the material in dist. water (1:100) before heating. A dilution in Eagle's MEM (1:100) or a previous dialysis against water does not have the same effect. This property was found in the following members of 4 DNA virus families: adenovirus 5, herpesvirus 1, SV 40 and bovine parvovirus 1. In contrast, members of pox- (vaccinia) and picornaviruses (polio 2, coxsackie B5) were found to be less thermostable under conditions of low ionic strength. Reovirus 3 showed no difference in inactivation. The observed effect may be of practical importance for heat disinfection of viruses and for their persistence in the environment. PMID:6271099

Wigand, R; Bachmann, P; Brandner, G

1981-01-01

128

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

PubMed

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

2013-12-27

129

Microbiology and management of neonatal necrotizing enterocolitis.  

PubMed

Necrotizing enterocolitis (NEC) is a clinical syndrome of ischemic necrosis of the bowel of multiple etiological factors that include the presence of intestinal ischemia, abnormal bacterial flora, and intestinal mucosal immaturity. Numerous reports have implied that the fecal microflora may contribute to the pathogenesis of NEC. A broad range of organisms generally found in the distal gastrointestinal tract have been recovered from the peritoneal cavity and blood of infants with NEC. The predominant organisms include Enterobacteriaceae (i.e., Escherichia coli, Klebsiella pneumoniae) , Clostridium spp., enteric pathogens (salmonellae, Coxsackie B2 virus, coronavirus, rotavirus), and potential pathogens (Bacteroides fragilis). The goals of the initial management is preventing ongoing damage, restoring hemostasis, and minimizing complications. Medical management includes withholding oral feeding, placement of nasogastric tube, abdominal decompression, paracentesis, vigorous intravenous hydration containing electrolytes and calories, support of the circulation, administration of antibiotics, and surveillance for deterioration or complications that require surgical intervention. Indications for surgery include clinical deterioration, perforation, peritonitis, obstruction, and abdominal mass. Prevention remains crucial to decrease the incidence of NEC. Preventive methods include cautious feeding regimens, the use of maternal breast milk, and the use of probiotics. PMID:18236362

Brook, Itzhak

2008-02-01

130

Isolation of Ether-Resistant Enteroviruses from Sewage: Methodology  

PubMed Central

Experiments were conducted to determine whether polio type 1 (Mahoney and coxsackie A8 viruses adsorb onto cotton fibers of sewer swabs. Negative results were obtained. It has been shown that viruses may exist in sewage as free virus particles or as bound (adsorbed) virus particles. The sewer-swab method of sampling is superior because it filters out the bound virus over several days; when collected, it represents a catch (grab) sample at that particular time which may or may not contain free virus. A simple method for the preparation of sewage inocula for virus isolations is described which samples the bound virus fraction. Only ether-resistant viruses can be isolated, and an ultracentrifuge is not required. By this method, an isolation rate between 60 and 80% of positive sewer swabs can be achieved. Corresponding figures of 84 and 96% were achieved by concentration of sewer-swab eluates with an ultracentrifuge. Quantitative studies showed that the virus concentration in raw sewage can be as high as one infectious particle per 0.5 ml.

Duff, Michael F.

1970-01-01

131

Protein levels of genes encoded on chromosome 21 in fetal Down syndrome brain: Challenging the gene dosage effect hypothesis (Part IV).  

PubMed

Down syndrome (DS) is the most frequent genetic disorder with mental retardation and caused by trisomy 21. Although the molecular mechanisms of the various phenotypes of DS could be due to overexpression of gene(s) on chromosome 21, several groups have challenged this gene dosage effect hypothesis. The near completion of the sequencing of human chromosome 21 provides unprecedented opportunities to understand the molecular pathology of DS, however, functional information on gene products is limited so far. We therefore evaluated the levels of six proteins whose genes are encoded on chromosome 21 (trefoil factor 1, trefoil factor 2, trefoil factor 3, coxsackie virus and adenovirus receptor, carbonyl reductase 1 and interferon- alpha receptor) in fetal cerebral cortex from DS and controls at the early second trimester using Western blot analysis. None of the investigated proteins showed overexpression in DS compared to controls suggesting that these proteins are not involved in abnormal development of fetal DS brain and that DS phenotype can not be simply explained by the gene dosage effect hypothesis. We are systematically quantifying all proteins whose genes are encoded on chromosome 21 and these studies may provide a better understanding of genotype-phenotype correlation in DS. PMID:12836057

Cheon, M S; Shim, K S; Kim, S H; Hara, A; Lubec, G

2003-07-01

132

Pinellia pedatisecta agglutinin targets drug resistant K562/ADR leukemia cells through binding with sarcolemmal membrane associated protein and enhancing macrophage phagocytosis.  

PubMed

Pinelliapedatisecta agglutinin (PPA) has previously been used in labeling fractions of myeloid leukemia cells in our laboratory. We report here that a bacterial expressed recombinant PPA domain b tagged with soluble coxsackie and adenovirus receptor (sCAR-PPAb) preferentially recognized drug resistant cancer cells K562/ADR and H460/5Fu, as compared to their parental cell lines. Pretreatment of K562/ADR cells with sCAR-PPAb significantly enhanced phagocytosis of K562/ADR by macrophages in vivo. Meanwhile, in a K562/ADR xenograft model, intratumoral injection of sCAR-PPAb induced macrophage infiltration and phagocytosis. Furthermore, immunoprecipitation, mass spectrometry and Western blot identified the membrane target of PPA on K562/ADR as sarcolemmal membrane associated protein (SLMAP). An antibody against SLMAP significantly promoted the phagocytosis of K562/ADR by macrophages in vitro. These findings suggest that PPA not only could be developed into a novel agent that can detect drug resistant cancer cells and predict chemotherapy outcome, but also it has potential value in immunotherapy against drug resistant cancer cells through inducing the tumoricidal activity of macrophages. PMID:24019967

Chen, Kan; Yang, Xinyan; Wu, Liqin; Yu, Meilan; Li, Xiaoyan; Li, Na; Wang, Shuanghui; Li, Gongchu

2013-01-01

133

Pinellia pedatisecta Agglutinin Targets Drug Resistant K562/ADR Leukemia Cells through Binding with Sarcolemmal Membrane Associated Protein and Enhancing Macrophage Phagocytosis  

PubMed Central

Pinelliapedatisecta agglutinin (PPA) has previously been used in labeling fractions of myeloid leukemia cells in our laboratory. We report here that a bacterial expressed recombinant PPA domain b tagged with soluble coxsackie and adenovirus receptor (sCAR-PPAb) preferentially recognized drug resistant cancer cells K562/ADR and H460/5Fu, as compared to their parental cell lines. Pretreatment of K562/ADR cells with sCAR-PPAb significantly enhanced phagocytosis of K562/ADR by macrophages in vivo. Meanwhile, in a K562/ADR xenograft model, intratumoral injection of sCAR-PPAb induced macrophage infiltration and phagocytosis. Furthermore, immunoprecipitation, mass spectrometry and Western blot identified the membrane target of PPA on K562/ADR as sarcolemmal membrane associated protein (SLMAP). An antibody against SLMAP significantly promoted the phagocytosis of K562/ADR by macrophages in vitro. These findings suggest that PPA not only could be developed into a novel agent that can detect drug resistant cancer cells and predict chemotherapy outcome, but also it has potential value in immunotherapy against drug resistant cancer cells through inducing the tumoricidal activity of macrophages.

Chen, Kan; Yang, Xinyan; Wu, Liqin; Yu, Meilan; Li, Xiaoyan; Li, Na; Wang, Shuanghui; Li, Gongchu

2013-01-01

134

Adsorption of enteroviruses to soil cores and their subsequent elution by artificial rainwater.  

PubMed

The adsorption and elution of a variety of human enteroviruses in a highly permeable, sandy soil was studied by using cores (43 by 125 mm) collected from an operating recharge basin on Long Island. Viruses studied included field and reference strains of polioviruses types 1 and 3 and reference strains of coxsackie virus B3 and echovirus types 1 and 6. Viruses suspended in treated sewage effluent were allowed to percolate through soil cores, and the filtrate was assayed for unadsorbed viruses. To determine the likelihood of desorption and mobilization, soil-bound viruses were subjected to a rinse with either treated sewage effluent or simulated rainwater which reflected the anion, cation, and pH characteristics of a typical northeastern United States rainfall. The results demonstrated that all polioviruses tested, including both reference and field strains, adsorbed extremely well to cores. Adsorption was somewhat reduced when clean, unconditioned soils were used. Soil-bound poliovirus strain LSc was not significantly mobilized by flooding columns with either a sewage effluent or rainwater rinse. One virus was mobilized by both types of rinses. The amount of viruses mobilized by rainwater rinses ranged from 24 to 66%. Variable adsorption-elution results were observed with other enteroviruses. Two guanidine-resistant mutants of poliovirus LSc demonstrated a soil adsorption-elution profile different from that of the parent strain. The data support the conclusion that soil adsorption-elution behavior is strain dependent and that poliovirus, particularly strain LSc, represents an inappropriate model. PMID:231936

Landry, E F; Vaughn, J M; Thomas, M Z; Beckwith, C A

1979-10-01

135

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

PubMed Central

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

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

2002-01-01

136

Human enterovirus 71 epidemics: what's next?  

PubMed Central

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

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

2013-01-01

137

Clathrin adaptor AP1B controls adenovirus infectivity of epithelial cells  

PubMed Central

Adenoviruses invading the organism via normal digestive or respiratory routes require the Coxsackie-adenovirus receptor (CAR) to infect the epithelial barrier cells. Because CAR is a component of tight junctions and the basolateral membrane and is normally excluded from the apical membrane, most epithelia are resistant to adenoviruses. However, we discovered that a specialized epithelium, the retinal pigment epithelium (RPE), anomalously expressed CAR at the apical surface and was highly susceptible to adenovirus infection. These properties of RPE cells correlated with the absence of the epithelial-specific clathrin adaptor AP1B. Furthermore, knockdown of this basolateral sorting adaptor in adenovirus-resistant MDCK cells promoted apical localization of CAR and increased dramatically Adenovirus infectivity. Targeting assays showed that AP1B is required for accurate basolateral recycling of CAR after internalization. AP1B knock down MDCK cells missorted CAR from recycling endosomes to the apical surface. In summary, we have characterized the cellular machinery responsible for normal sorting of an adenovirus receptor and illustrated how tissue-specific variations in such machinery result in drastic changes in tissue-susceptibility to adenoviruses.

Diaz, Fernando; Gravotta, Diego; Deora, Ami; Schreiner, Ryan; Schoggins, John; Falck-Pedersen, Erik; Rodriguez-Boulan, Enrique

2009-01-01

138

Cytokine and Chemokine Production by Human Pancreatic Islets Upon Enterovirus Infection  

PubMed Central

Enteroviruses of the human enterovirus B species (HEV-Bs) (e.g., coxsackie B viruses [CVBs] and echoviruses) have been implicated as environmental factors that trigger/accelerate type 1 diabetes, but the underlying mechanism remains elusive. The aim of this study was to gain insight into the cytokines and chemokines that are produced by human pancreatic islets upon infection with CVBs. To this end, we studied the response of human islets of Langerhans upon mock or CVB3 infection. Using quantitative PCR, we showed that upon CVB3 infection, transcription of interferon (IFN), IFN-stimulated genes, and inflammatory genes was induced. Analysis of secreted cytokines and chemokines by Luminex technology confirmed production and secretion of proinflammatory cytokines (e.g., interleukin [IL]-6 and tumor necrosis factor-?) as well as various chemotactic proteins, such as IFN-?–induced protein 10, macrophage inflammatory protein (MIP)-1?, MIP-1?, and IL-8. Infection with other HEV-Bs induced similar responses, yet their extent depended on replication efficiency. Ultra violet–inactivated CVB3 did not induce any response, suggesting that virus replication is a prerequisite for antiviral responses. Our data represent the first comprehensive overview of inflammatory mediators that are secreted by human islets of Langerhans upon CVB infection and may shed light on the role of enteroviruses in type 1 diabetes pathogenesis.

Schulte, Barbara M.; Lanke, Kjerstin H.W.; Piganelli, Jon D.; Kers-Rebel, Esther D.; Bottino, Rita; Trucco, Massimo; Huijbens, Richard J.F.; Radstake, Timothy R.D.J.; Engelse, Marten A.; de Koning, Eelco J.P.; Galama, Jochem M.; Adema, Gosse J.; van Kuppeveld, Frank J.M.

2012-01-01

139

Phagocytosis of Picornavirus-Infected Cells Induces an RNA-Dependent Antiviral State in Human Dendritic Cells?  

PubMed Central

Dendritic cells (DCs) play a central role in instructing antiviral immune responses. DCs, however, can become targeted by different viruses themselves. We recently demonstrated that human DCs can be productively infected with echoviruses (EVs), but not coxsackie B viruses (CVBs), both of which are RNA viruses belonging to the Enterovirus genus of the Picornaviridae family. We now show that phagocytosis of CVB-infected, type I interferon-deficient cells induces an antiviral state in human DCs. Uptake of infected cells increased the expression of the cytoplasmic RNA helicases retinoic acid-inducible gene I and melanoma differentiation-associated gene 5 as well as other interferon-stimulated genes and protected DCs against subsequent infection with EV9. These effects depended on recognition of viral RNA and could be mimicked by exposure to the synthetic double-stranded RNA analogue poly(I:C) but not other Toll-like receptor (TLR) ligands. Blocking endosomal acidification abrogated protection, suggesting a role for TLRs in the acquisition of an antiviral state in DCs. In conclusion, recognition of viral RNA rapidly induces an antiviral state in human DCs. This might provide a mechanism by which DCs protect themselves against viruses when attracted to an environment with ongoing infection.

Kramer, Matthijs; Schulte, Barbara M.; Toonen, Liza W. J.; Barral, Paola M.; Fisher, Paul B.; Lanke, Kjerstin H. W.; Galama, Jochem M. D.; van Kuppeveld, Frank J. M.; Adema, Gosse J.

2008-01-01

140

Cytokine and chemokine production by human pancreatic islets upon enterovirus infection.  

PubMed

Enteroviruses of the human enterovirus B species (HEV-Bs) (e.g., coxsackie B viruses [CVBs] and echoviruses) have been implicated as environmental factors that trigger/accelerate type 1 diabetes, but the underlying mechanism remains elusive. The aim of this study was to gain insight into the cytokines and chemokines that are produced by human pancreatic islets upon infection with CVBs. To this end, we studied the response of human islets of Langerhans upon mock or CVB3 infection. Using quantitative PCR, we showed that upon CVB3 infection, transcription of interferon (IFN), IFN-stimulated genes, and inflammatory genes was induced. Analysis of secreted cytokines and chemokines by Luminex technology confirmed production and secretion of proinflammatory cytokines (e.g., interleukin [IL]-6 and tumor necrosis factor-?) as well as various chemotactic proteins, such as IFN-?-induced protein 10, macrophage inflammatory protein (MIP)-1?, MIP-1?, and IL-8. Infection with other HEV-Bs induced similar responses, yet their extent depended on replication efficiency. Ultra violet-inactivated CVB3 did not induce any response, suggesting that virus replication is a prerequisite for antiviral responses. Our data represent the first comprehensive overview of inflammatory mediators that are secreted by human islets of Langerhans upon CVB infection and may shed light on the role of enteroviruses in type 1 diabetes pathogenesis. PMID:22596052

Schulte, Barbara M; Lanke, Kjerstin H W; Piganelli, Jon D; Kers-Rebel, Esther D; Bottino, Rita; Trucco, Massimo; Huijbens, Richard J F; Radstake, Timothy R D J; Engelse, Marten A; de Koning, Eelco J P; Galama, Jochem M; Adema, Gosse J; van Kuppeveld, Frank J M

2012-08-01

141

Phagocytosis of picornavirus-infected cells induces an RNA-dependent antiviral state in human dendritic cells.  

PubMed

Dendritic cells (DCs) play a central role in instructing antiviral immune responses. DCs, however, can become targeted by different viruses themselves. We recently demonstrated that human DCs can be productively infected with echoviruses (EVs), but not coxsackie B viruses (CVBs), both of which are RNA viruses belonging to the Enterovirus genus of the Picornaviridae family. We now show that phagocytosis of CVB-infected, type I interferon-deficient cells induces an antiviral state in human DCs. Uptake of infected cells increased the expression of the cytoplasmic RNA helicases retinoic acid-inducible gene I and melanoma differentiation-associated gene 5 as well as other interferon-stimulated genes and protected DCs against subsequent infection with EV9. These effects depended on recognition of viral RNA and could be mimicked by exposure to the synthetic double-stranded RNA analogue poly(I:C) but not other Toll-like receptor (TLR) ligands. Blocking endosomal acidification abrogated protection, suggesting a role for TLRs in the acquisition of an antiviral state in DCs. In conclusion, recognition of viral RNA rapidly induces an antiviral state in human DCs. This might provide a mechanism by which DCs protect themselves against viruses when attracted to an environment with ongoing infection. PMID:18184700

Kramer, Matthijs; Schulte, Barbara M; Toonen, Liza W J; Barral, Paola M; Fisher, Paul B; Lanke, Kjerstin H W; Galama, Jochem M D; van Kuppeveld, Frank J M; Adema, Gosse J

2008-03-01

142

Combined transductional untargeting/retargeting and transcriptional restriction enhance adenovirus gene targeting and therapy for hepatic colorectal cancer tumors  

PubMed Central

Unresectable hepatic colorectal cancer (CRC) metastases are a leading cause of cancer mortality. These tumors, and other epithelial tumors, often express both cyclooxygenase 2 (COX-2) and carcinoembryonic antigen (CEA). Because adenovirus vectors infect liver and lack tumor tropism, they cannot be utilized for systemic therapy of hepatic metastases. We used COX-2 transcriptional restriction, in combination with transductional adenovirus hepatic untargeting and tumor retargeting by a bispecific adapter, sCARhMFE, composed of sCAR (the Coxsackie and Adenovirus Receptor ectodomain) and MFE-23 (a single-chain anti-CEA antibody), to untarget liver following intravenous administration of adenovirus vectors expressing firefly luciferase and to retarget virus to hepatic colorectal tumor xenografts and non-small cell lung tumor xenografts. To improve both liver untargeting and tumor retargeting, we developed sCARfMFE, a trimerized sCARhMFE adapter. Trimerization greatly improves both untargeting of CAR-dependent adenovirus infection and CEA-dependent virus retargeting, in culture and in vivo. Combining sCARfMFE bispecific adapter transductional liver untargeting and transductional tumor retargeting with COX-2 transcriptional tumor-restricted transgene expression increases systemically-administered adenovirus therapeutic efficacy for hepatic CRC tumors, using Herpes Virus Type 1 thymidine kinase as a therapeutic gene in conjunction with the prodrug ganciclovir. Both transductional untargeting and COX-2 transcriptional restriction also reduce HSV1-tk/GCV hepatic toxicity. In addition, transductional sCARfMFE untargeting reduces the innate immune response to systemic adenovirus administration. Combined transductional liver Ad untargeting, transductional tumor retargeting and transcriptional transgene restriction suggests a means to engineer practical, effective therapeutic agents for hepatic CRC metastases in particular, as well as hepatic metastases of other epithelial cancers.

Li, Hua-Jung; Everts, Maaike; Yamamoto, Masato; Curiel, David T.; Herschman, Harvey R.

2009-01-01

143

Robust infectivity and replication of Delta-24 adenovirus induce cell death in human medulloblastoma.  

PubMed

The diverse advanced treatment modalities currently available to children with medulloblastoma, including surgery and radiotherapy, are associated with deleterious side effects and often with an unfavorable prognosis. A mutant adenovirus, Delta-24, which has a 24-base pair deletion in the Rb-binding region of the E1A gene, demonstrates selective replication and oncolysis in various malignant phenotypes. Here we report the ability of Delta-24 to kill medulloblastoma cells. Flow cytometric analyses of cell receptors demonstrated expression of the coxsackie adenovirus receptor and RGD-related integrins in the assessed medulloblastoma cell lines. Infectivity assays using a replication-deficient adenovirus to transduce the green fluorescence protein gene showed that the Delta-24 adenovirus infects 99% of Daoy and 46% of D283 Med medulloblastoma cells at a multiplicity of infection (MOI) of 50. Within 4 days after infecting medulloblastoma cells with Delta-24, a noticeable cytopathic effect was produced. Delta-24 induced a total cytopathic effect in Daoy and D283 Med medulloblastoma cells after 6 and 8 days of infection, respectively. In the infected population of cells, cell death correlated with the accumulation of cells in the S phase. At 5 days post-infection with 2.5 MOIs of Delta-24 adenovirus, the percentage of Daoy medulloblastoma cells in the S phase increased to 71.9+/-5.5%, compared with control values of 20.5+/-1.4%. The release of viral progeny was quantified as being increased by two orders of magnitude, indicating efficient replication of Delta-24 in medulloblastoma cells. This is the first report of the ability of oncolytic adenoviruses to infect and kill medulloblastoma cells, the findings of which suggest the potential efficacy of Delta-24 as a therapy for human medulloblastoma tumors. PMID:15332115

Stolarek, Robert; Gomez-Manzano, Candelaria; Jiang, Hong; Suttle, Gary; Lemoine, Michael G; Fueyo, Juan

2004-11-01

144

The effect of surface modification of adenovirus with an arginine-grafted bioreducible polymer on transduction efficiency and immunogenicity in cancer gene therapy.  

PubMed

Adenoviral vectors offer many advantages for cancer gene therapy, including high transduction efficiency, but safety concerns related to severe immunogenicity and other side effects have led to careful reconsideration of their use in human clinical trials. To overcome these issues, a strategy of generating hybrid vectors that combine viral and non-viral elements as more intelligent gene carriers has been employed. Here, we coated adenovirus (Ad) with an arginine-grafted bioreducible polymer (ABP) via electrostatic interaction. We examined the effect of ABP-coated Ad complex at various ABP molecules/Ad particle ratios. Enhanced transduction efficiency was observed in cells treated with cationic ABP polymer-coated Ad complex compared to naked Ad. We also examined the coating of Ad with ABP polymers at the optimal polymer ratio using dynamic light scattering and transmission electron microscopy. In both high and low coxsackie virus and adenovirus receptor (CAR)-expressing cells, ABP-coated Ad complex produced higher levels of transgene expression than cationic polymer 25K PEI. Notably, high cytotoxicity was observed with 25K PEI-coated Ad complex treatment, but not with ABP-coated Ad complex treatment. In addition, ABP-coated Ad complex was not significantly inhibited by serum, in contrast to naked Ad. Moreover, ABP-coated Ad complex significantly reduced the innate immune response relative to naked Ad, as assessed by interleukin-6 (IL-6) cytokine release from macrophage cells. Overall, our studies demonstrate that Ad complex formed with ABP cationic polymer may improve the efficiency of Ad and be a promising tool for cancer gene therapy. PMID:19962189

Kim, Pyung-Hwan; Kim, Tae-Il; Yockman, James W; Kim, Sung Wan; Yun, Chae-Ok

2010-03-01

145

Identification of HI-Like Loop in CELO Adenovirus Fiber for Incorporation of Receptor Binding Motifs?  

PubMed Central

Vectors based on the chicken embryo lethal orphan (CELO) avian adenovirus (Ad) have two attractive properties for gene transfer applications: resistance to preformed immune responses to human Ads and the ability to grow in chicken embryos, allowing low-cost production of recombinant viruses. However, a major limitation of this technology is that CELO vectors demonstrate decreased efficiency of gene transfer into cells expressing low levels of the coxsackie-Ad receptor (CAR). In order to improve the efficacy of gene transfer into CAR-deficient cells, we modified viral tropism via genetic alteration of the CELO fiber 1 protein. The ?v integrin-binding motif (RGD) was incorporated at two different sites of the fiber 1 knob domain, within an HI-like loop that we identified and at the C terminus. Recombinant fiber-modified CELO viruses were constructed containing secreted alkaline phosphatase (SEAP) and enhanced green fluorescent protein genes as reporter genes. Our data show that insertion of the RGD motif within the HI-like loop of the fiber resulted in significant enhancement of gene transfer into CAR-negative and CAR-deficient cells. In contrast, CELO vectors containing the RGD motif at the fiber 1 C terminus showed reduced transduction of all cell lines. CELO viruses modified with RGD at the HI-like loop transduced the SEAP reporter gene into rabbit mammary gland cells in vivo with an efficiency significantly greater than that of unmodified CELO vector and similar to that of Ad type 5 vector. These results illustrate the potential for efficient CELO-mediated gene transfer into a broad range of cell types through modification of the identified HI-like loop of the fiber 1 protein.

Logunov, Denis Y.; Zubkova, Olga V.; Karyagina-Zhulina, Anna S.; Shuvalova, Eugenia A.; Karpov, Andrei P.; Shmarov, Maxim M.; Tutykhina, Irina L.; Alyapkina, Yulia S.; Grezina, Natalia M.; Zinovieva, Natalia A.; Ernst, Lev K.; Gintsburg, Alexsandr L.; Naroditsky, Boris S.

2007-01-01

146

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

PubMed Central

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

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

2014-01-01

147

Transduction of Brain Dopamine Neurons by Adenoviral Vectors Is Modulated by CAR Expression: Rationale for Tropism Modified Vectors in PD Gene Therapy  

PubMed Central

Background Gene-based therapy is a new paradigm for the treatment of Parkinson disease (PD) and offers considerable promise for precise targeting and flexibility to impact multiple pathobiological processes for which small molecule agents are not available. Some success has been achieved utilizing adeno-associated virus for this approach, but it is likely that the characteristics of this vector system will ultimately create barriers to progress in clinical therapy. Adenovirus (Ad) vector overcomes limitations in payload size and targeting. The cellular tropism of Ad serotype 5 (Ad5)–based vectors is regulated by the Ad attachment protein binding to its primary cellular receptor, the coxsackie and adenovirus receptor (CAR). Many clinically relevant tissues are refractory to Ad5 infection due to negligible CAR levels but can be targeted by tropism-modified, CAR-independent forms of Ad. Our objective was to evaluate the role of CAR protein in transduction of dopamine (DA) neurons in vivo. Methodology/Principal Findings Ad5 was delivered to the substantia nigra (SN) in wild type (wt) and CAR transgenic animals. Cellular tropism was assessed by immunohistochemistry (IHC) in the SN and striatal terminals. CAR expression was assessed by western blot and IHC. We found in wt animals, Ad5 results in robust transgene expression in astrocytes and other non-neuronal cells but poor infection of DA neurons. In contrast, in transgenic animals, Ad5 infects SNc neurons resulting in expression of transduced protein in their striatal terminals. Western blot showed low CAR expression in the ventral midbrain of wt animals compared to transgenic animals. Interestingly, hCAR protein localizes with markers of post-synaptic structures, suggesting synapses are the point of entry into dopaminergic neurons in transgenic animals. Conclusions/Significance These findings demonstrate that CAR deficiency limits infection of wild type DA neurons by Ad5 and provide a rationale for the development of tropism-modified, CAR-independent Ad-vectors for use in gene therapy of human PD.

Lewis, Travis B.; Glasgow, Joel N.; Glandon, Anya M.; Curiel, David T.; Standaert, David G.

2010-01-01

148

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

PubMed Central

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

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

2013-01-01

149

A fiber-modified adenovirus co-expressing HSV-TK and Coli.NTR enhances antitumor activities in breast cancer cells  

PubMed Central

Breast cancers especially in late and metastatic stages remain refractory to treatment despite advances in surgical techniques and chemotherapy. Suicide gene therapy based on adenoviral technology will be promising strategies for such advanced diseases. We previously showed that co-expression of herpes simplex virus thymidine kinase (HSV-TK) and Escherichia coli nitroreductase (Coli.NTR) by an hTERT-driven adenovirus vector resulted in additive anti-tumor effects in breast cancer cells in vitro and in vivo. As many tumor tissue and cancer cells express low level of coxsackie-adenovirus receptor (CAR), which is the functional receptor for the fiber protein of human adenovirus serotype 5 (Ad5), novel Ad5 vectors containing genetically modifi ed fiber are attractive vehicles for achieving targeted gene transfer and improving suicide gene expression in these cancer cells. In the present study, we first built a simplified Ad5 vector platform for fiber modification and quick detection for gene transfer. Then a fiber-modified adenovirus vector containing an RGD motif in the HI loop of the fiber knob was constructed. After recombined with HSV-TK and Coli.NTR gene, this fiber-modified Ad5 vector (Ad-RGD-hT-TK/NTR) was compared with that of our previously constructed Ad5 vector (Ad-hT-TK/NTR) for its therapeutic effects in human breast cancer cell lines. The anti-tumor activity of Ad-RGD-hT-TK/NTR was significantly enhanced compared with Ad-hT-TK/NTR both in vitro and in vivo. This new vector platform provided a robust and simplified approach for capsid modification, and the fiber-modified Ad5 with double suicide genes under the control of hTERT promoter would be a useful gene therapy strategy for breast cancer.

Zhan, Yang; Yu, Bin; Wang, Zhen; Zhang, Yu; Zhang, Hai-Hong; Wu, Hao; Feng, Xiao; Geng, Ran-Shen; Kong, Wei; Yu, Xiang-Hui

2014-01-01

150

[Effect of IL-17A on levels of antiheart autoantibodies in mice with viral myocarditis].  

PubMed

Objective To explore the effect of interleukin-17A (IL-17A) on the serum level of antiheart autoantibodies in mice with viral myocarditis. Methods Male wild-type (WT) and IL-17A-deficient (IL-17A(-/-)) BALB/c mice were intraperitoneally injected with Coxsackie virus B3 (CVB3) for establishing VMC models (VMC-WT group and VMC-IL-17A(-/-) group). Meanwhile, a control group (WT group) of WT mice were established by i.p. administration of phosphate buffered saline (PBS). Paraffin sections of cardiac tissues were made 14 days after CVB3 injection. Myocardial histopathologic changes were evaluated by HE staining. The levels of anti-adenine nucleotide translocator (ANT) autoantibody, anti-?-myosin heavy chain (?-MHC) autoantibody and anti-cardiac L-type calcium channel (CACH2) autoantibody in sera were measured by ELISA. Results Compared with WT group, the levels of anti-ANT-autoantibody and anti-?-MHC-autoantibody significantly increased in VMC-WT group (P<0.01, P<0.05), while the concentration of anti-CACH2-autoantibody showed no significant difference between WT and VMC-WT groups (P>0.05). Compared with VMC-WT group, the level of anti-ANT-autoantibody was reduced in VMC-IL-17A(-/-) group (P<0.05), while the levels of anti-?-MHC-autoantibody and anti-CACH2-autoantibody showed no significant difference between them (P>0.05). Conclusion IL-17A contributed to the secretion of anti-ANT-autoantibody of VMC mice, but had no effect on the secretion of anti-?-MHC-autoantibody and anti-CACH2-autoantibody in VMC mice. PMID:24909282

Lai, Wenying; Wu, Weifeng; Pan, Xiaofen; Kong, Qing

2014-06-01

151

Possible involvement of Opa-interacting protein 5 in adipose proliferation and obesity.  

PubMed

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

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

2014-01-01

152

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

PubMed Central

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

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

2014-01-01

153

Proteomic and biochemical analyses reveal the activation of unfolded protein response, ERK-1/2 and ribosomal protein S6 signaling in experimental autoimmune myocarditis rat model  

PubMed Central

Background To investigate the molecular and cellular pathogenesis underlying myocarditis, we used an experimental autoimmune myocarditis (EAM)-induced heart failure rat model that represents T cell mediated postinflammatory heart disorders. Results By performing unbiased 2-dimensional electrophoresis of protein extracts from control rat heart tissues and EAM rat heart tissues, followed by nano-HPLC-ESI-QIT-MS, 67 proteins were identified from 71 spots that exhibited significantly altered expression levels. The majority of up-regulated proteins were confidently associated with unfolded protein responses (UPR), while the majority of down-regulated proteins were involved with the generation of precursor metabolites and energy metabolism in mitochondria. Although there was no difference in AKT signaling between EAM rat heart tissues and control rat heart tissues, the amounts and activities of extracellular signal-regulated kinase (ERK)-1/2 and ribosomal protein S6 (rpS6) were significantly increased. By comparing our data with the previously reported myocardial proteome of the Coxsackie viruses of group B (CVB)-mediated myocarditis model, we found that UPR-related proteins were commonly up-regulated in two murine myocarditis models. Even though only two out of 29 down-regulated proteins in EAM rat heart tissues were also dysregulated in CVB-infected rat heart tissues, other proteins known to be involved with the generation of precursor metabolites and energy metabolism in mitochondria were also dysregulated in CVB-mediated myocarditis rat heart tissues, suggesting that impairment of mitochondrial functions may be a common underlying mechanism of the two murine myocarditis models. Conclusions UPR, ERK-1/2 and S6RP signaling were activated in both EAM- and CVB-induced myocarditis murine models. Thus, the conserved components of signaling pathways in two murine models of acute myocarditis could be targets for developing new therapeutic drugs or methods aimed at treating enigmatic myocarditis.

2011-01-01

154

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

PubMed

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 (123)I-scintigraphy. Reduction of adenovirus liver pooling resulted in decreased hepatotoxicity and increased transduction efficiency in peripheral xenograft tumors. (124)I-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 (131)I. 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.Molecular Therapy-Nucleic Acids (2013) 2, e131; doi:10.1038/mtna.2013.58; published online 5 November 2013. 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

2013-01-01

155

Abnormal expression of multiple proteins predicts cancer-specific mortality in patients with high-grade non-muscle-invasive bladder cancer treated with transurethral resection  

PubMed Central

High-grade non-muscle-invasive bladder urothelial carcinoma leads to various outcomes. It can cause death even after radical cystectomy and is treated only by transurethral resection (TUR). In the present study, we aimed to determine whether the molecular markers E-cadherin, coxsackie adenovirus receptor (CAR), S100A4 and uroplakin III are associated with clinicopathological outcomes in patients with high-grade non-muscle invasive bladder cancer (NMIBC) treated with TUR. Immunohistochemical staining was performed on serial sections from specimens obtained from 77 patients. Expression patterns were stratified according to the number of abnormally expressed markers: 0–1 or ?2. The median follow-up time was 56 months (range, 3–287). The results from the present study indicated that expression of E-cadherin, CAR, S100A4 and uroplakin III was abnormal in 16, 17, 27 and 61% of tumors, respectively. Results of the log-rank test revealed that patients with abnormal expression of multiple molecular markers had a significantly increased risk of bladder cancer-specific mortality (P=0.016). The 5-year cancer-specific survival rates were 91 and 66% for patients with 0–1 and ?2 molecular markers, respectively. No individual marker was associated with disease prognosis. Multivariate models that included clinicopathological outcomes and classified molecular markers indicated that abnormal expression of multiple molecular markers and lack of bacillus Calmette-Guérin (BCG) instillation are predictors of cancer-specific death (P=0.046 and 0.029, respectively). Abnormal expression of multiple molecular markers is a strong predictor of mortality in bladder cancer patients undergoing TUR, suggesting that high-grade non-muscle-invasive cancer is characterized by a variety of pathophysiological pathways. A combination of molecular markers may be useful in a minimally invasive modality for determining prognosis.

TSUMURA, HIDEYASU; MATSUMOTO, KAZUMASA; SATO, YUICHI; IKEDA, MASAOMI; FUJITA, TETSUO; SATOH, TAKEFUMI; IWAMURA, MASATSUGU

2013-01-01

156

Enterovirus infection among patients admitted to hospital in Hong Kong in 2010: epidemiology, clinical characteristics, and importance of molecular diagnosis.  

PubMed

The activity of hand, foot, and mouth disease (HFMD) in Hong Kong was high in 2010. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) had been performed routinely for diagnosis of enterovirus (EV) infection among hospitals in a geographical cluster. The aim of the study was to describe the epidemiology and the clinical pattern of EV-related hospital admission in 2010, and evaluate the impact of RT-PCR compared to conventional method. This was a retrospective study and patients with laboratory confirmed EV infection were included. Demographic information, clinical features, complications, and laboratory findings were analyzed. Among the 113 patients identified, HFMD was the most common presentation (84/113, 74%), followed by meningitis (10/113, 9%). Respiratory (52/113, 46%) and gastrointestinal symptoms like vomiting (36/113, 32%) and diarrhea (15/113, 13%) were quite common. For cases with central nervous system (CNS) or myocardial complications, there were significantly fewer oral ulcer and rash over hands and feet compared to their counter group. Serious infection mainly affected children younger than 1-year old and adults. Dual infection was noted among seven patients (6%). Compared to RT-PCR, conventional virus isolation detected only 14-16% of the infections. The relative low culture positive rate could be explained by the circulation of Coxsackie A6 in 2010 which was difficult to be isolated by cell culture. Diagnosis of EV infection among hospitalized patients may not be straightforward. EV RT-PCR significantly improved laboratory diagnosis and delineation of epidemiology of EV infection compared to conventional methods. PMID:23824683

Lee, May Kin-Ping; Chan, Paul Kay-Sheung; Ho, Iok-Ieng Yolanda; Lai, Wai-Man Raymond

2013-10-01

157

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.

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

2003-01-01

158

Multimodality Diagnostic Imaging in Unilateral Acute Idiopathic Maculopathy  

PubMed Central

Objective To describe the clinical features and imaging characteristics in unilateral acute idiopathic maculopathy (UAIM). Methods This is a retrospective review of four patients diagnosed with UAIM. Clinical characteristics (age, symptoms, Snellen visual acuity (VA), and funduscopic features) and images from spectral-domain optical coherence tomography (SD-OCT), fundus autofluorescence (FAF), fluorescein (FA), and indocyanine green (ICG) angiography were analyzed. Results The median age at presentation was 31 years (range 27–52 years). The median interval between symptom onset and presentation was four weeks (range 1–20 weeks). Associated systemic findings included a viral prodrome (50%), orchitis (50%), hand-foot-mouth disease (25%), and positive Coxsackie virus titers (50%). The median presenting VA was 20/400 (range 20/70–1/400), which improved to 20/30 (range 20/20–20/60) at final follow-up. The median follow-up time was 6 weeks (range 0–8 weeks). Early in the disease course, the central macula developed irregular, circular areas of white-grey discoloration. Following recovery, the macula had a stippled retinal pigment epithelium characterized by rarefaction and hyperplasia. FA demonstrated irregular early hyperfluorescence and late subretinal hyperfluorescence. SD-OCT showed a partially reversible disruption of the outer photoreceptor layer. FAF initially revealed stippled autofluorescence that eventually became more hypoautofluorescent. ICG showed “moth-eaten” appearing choroidal vasculature, suggestive of choroidal inflammation. Conclusions The imaging characteristics highlight the structural changes during the active and resolution phases of UAIM. The visual recovery correlates with structural changes and suggests that the pathogenesis involves inflammation of the inner choroid, retinal pigment epithelium, and outer photoreceptor complex that is partially reversible.

Jung, Cecilia S.; Payne, John F.; Bergstrom, Chris S.; Cribbs, Blaine E.; Yan, Jiong; Hubbard, G. Baker; Olsen, Timothy W.; Yeh, Steven

2014-01-01

159

Magnetically Responsive Biodegradable Nanoparticles Enhance Adenoviral Gene Transfer in Cultured Smooth Muscle and Endothelial Cells  

PubMed Central

Replication-defective adenoviral (Ad) vectors have shown promise as a tool for gene delivery-based therapeutic applications. Their clinical use is however limited by therapeutically suboptimal transduction levels in cell types expressing low levels of Coxsackie-Ad receptor (CAR), the primary receptor responsible for the cell entry of the virus, and by systemic adverse reactions. Targeted delivery achievable with Ad complexed with biodegradable magnetically responsive nanoparticles (MNP) may therefore be instrumental for improving both the safety and efficiency of these vectors. Our hypothesis was that magnetically driven delivery of Ad affinity-bound to biodegradable MNP can substantially increase transgene expression in CAR deficient vascular cells in culture. Fluorescently labeled MNP were formulated from polylactide with inclusion of iron oxide and surface-modified with the D1 domain of CAR as an affinity linker. MNP cellular uptake and GFP reporter transgene expression were assayed fluorimetrically in cultured endothelial and smooth muscle cells using ?ex/?em of 540 nm/575 nm and 485 nm/535 nm, respectively. Stable vector-specific association of Ad with MNP resulted in formation of MNP–Ad complexes displaying rapid cell binding kinetics following a brief exposure to a high gradient magnetic field with resultant gene transfer levels significantly increased compared to free vector or nonmagnetic control treatment. Multiple regression analysis suggested a mechanism of MNP–Ad mediated transduction distinct from that of free Ad, and confirmed the major contribution of the complexes to the gene transfer under magnetic conditions. The magnetically enhanced transduction was achieved without compromising the cell viability or growth kinetics. The enhancement of adenoviral gene delivery by affinity complexation with biodegradable MNP represents a promising approach with a potential to extend the applicability of the viral gene therapeutic strategies.

Chorny, Michael; Fishbein, Ilia; Alferiev, Ivan; Levy, Robert J.

2012-01-01

160

Rotavirus-specific T cell responses and cytokine mRNA expression in children with diabetes-associated autoantibodies and type 1 diabetes.  

PubMed

Rotavirus infections have been implicated as a possible trigger of type 1 diabetes. We elucidated this connection by comparing peripheral blood T cell responses to rotavirus between children with newly diagnosed type 1 diabetes (n = 43), healthy children with multiple diabetes-associated autoantibodies (n = 36) and control children carrying human leukocyte antigen (HLA)-conferred susceptibility to type 1 diabetes but without autoantibodies (n = 104). Lymphocyte proliferation assays based on stimulation with an antigen were performed using freshly isolated peripheral blood mononuclear cells (PBMC) and IgG and IgA class rotavirus antibodies were measured using plasma samples collected from the children. The expression of interferon (IFN)-gamma, interleukin (IL)-4, IL-10 and transforming growth factor (TGF)-beta in PBMC was studied with real-time polymerase chain reaction (PCR) in a subgroup of 38 children. No differences were observed in the strength or frequency of positive T cell responses to rotavirus between children with overt diabetes, children with multiple autoantibodies and control children. Children with diabetes-associated autoantibodies had, instead, stronger T cell responses to purified coxsackie B4 virus than control children. Rotavirus-stimulated lymphocytes from autoantibody-positive children produced more IL-4 and phytohaemagglutinin (PHA)-stimulated lymphocytes more IL-4 and IFN-gamma than lymphocytes from control children. PHA-stimulated lymphocytes from children with diabetes also produced more IL-4 and purified protein derivative (PPD)-stimulated lymphocytes less TGF-beta than lymphocytes from autoantibody-negative control children. In conclusion, our lymphocyte proliferation studies did not provide evidence supporting an association between rotavirus infections and the development of type 1 diabetes or diabetes-associated autoantibodies in young children. PMID:16879245

Mäkelä, M; Oling, V; Marttila, J; Waris, M; Knip, M; Simell, O; Ilonen, J

2006-08-01

161

In vitro culture of embryonic mouse intestinal epithelium: cell differentiation and introduction of reporter genes  

PubMed Central

Background Study of the normal development of the intestinal epithelium has been hampered by a lack of suitable model systems, in particular ones that enable the introduction of exogenous genes. Production of such a system would advance our understanding of normal epithelial development and help to shed light on the pathogenesis of intestinal neoplasia. The criteria for a reliable culture system include the ability to perform real time observations and manipulations in vitro, the preparation of wholemounts for immunostaining and the potential for introducing genes. Results The new culture system involves growing mouse embryo intestinal explants on fibronectin-coated coverslips in basal Eagle's medium+20% fetal bovine serum. Initially the cultures maintain expression of the intestinal transcription factor Cdx2 together with columnar epithelial (cytokeratin 8) and mesenchymal (smooth muscle actin) markers. Over a few days of culture, differentiation markers appear characteristic of absorptive epithelium (sucrase-isomaltase), goblet cells (Periodic Acid Schiff positive), enteroendocrine cells (chromogranin A) and Paneth cells (lysozyme). Three different approaches were tested to express genes in the developing cultures: transfection, electroporation and adenoviral infection. All could introduce genes into the mesenchyme, but only to a small extent into the epithelium. However the efficiency of adenovirus infection can be greatly improved by a limited enzyme digestion, which makes accessible the lateral faces of cells bearing the Coxsackie and Adenovirus Receptor. This enables reliable delivery of genes into epithelial cells. Conclusion We describe a new in vitro culture system for the small intestine of the mouse embryo that recapitulates its normal development. The system both provides a model for studying normal development of the intestinal epithelium and also allows for the manipulation of gene expression. The explants can be cultured for up to two weeks, they form the full repertoire of intestinal epithelial cell types (enterocytes, goblet cells, Paneth cells and enteroendocrine cells) and the method for gene introduction into the epithelium is efficient and reliable.

Quinlan, Jonathan M; Yu, Wei-Yuan; Hornsey, Mark A; Tosh, David; Slack, Jonathan MW

2006-01-01

162

Phagocytosis of Enterovirus-Infected Pancreatic ?-Cells Triggers Innate Immune Responses in Human Dendritic Cells  

PubMed Central

OBJECTIVE Type 1 diabetes is a chronic endocrine disorder in which enteroviruses, such as coxsackie B viruses and echoviruses, are possible environmental factors that can trigger or accelerate disease. The development or acceleration of type 1 diabetes depends on the balance between autoreactive effector T-cells and regulatory T-cells. This balance is particularly influenced by dendritic cells (DCs). The goal of this study was to investigate the interaction between enterovirus-infected human pancreatic islets and human DCs. RESEARCH DESIGN AND METHODS In vitro phagocytosis of human or porcine primary islets or Min6 mouse insuloma cells by DCs was investigated by flow cytometry and confocal analysis. Subsequent innate DC responses were monitored by quantitative PCR and Western blotting of interferon-stimulated genes (ISGs). RESULTS In this study, we show that both mock- and coxsackievirus B3 (CVB3)-infected human and porcine pancreatic islets were efficiently phagocytosed by human monocyte–derived DCs. Phagocytosis of CVB3-infected, but not mock-infected, human and porcine islets resulted in induction of ISGs in DCs, including the retinoic acid–inducible gene (RIG)-I–like helicases (RLHs), RIG-I, and melanoma differentiation–associated gene 5 (Mda5). Studies with murine Min6 insuloma cells, which were also efficiently phagocytosed, revealed that increased ISG expression in DCs upon encountering CVB-infected cells resulted in an antiviral state that protected DCs from subsequent enterovirus infection. The observed innate antiviral responses depended on RNA within the phagocytosed cells, required endosomal acidification, and were type I interferon dependent. CONCLUSIONS Human DCs can phagocytose enterovirus-infected pancreatic cells and subsequently induce innate antiviral responses, such as induction of RLHs. These responses may have important consequences for immune homeostasis in vivo and may play a role in the etiology of type 1 diabetes.

Schulte, Barbara M.; Kramer, Matthijs; Ansems, Marleen; Lanke, Kjerstin H.W.; van Doremalen, Neeltje; Piganelli, Jon D.; Bottino, Rita; Trucco, Massimo; Galama, Jochem M.D.; Adema, Gosse J.; van Kuppeveld, Frank J.M.

2010-01-01

163

Chemoprevention gene therapy (CGT) of pancreatic cancer using perillyl alcohol and a novel chimeric serotype cancer terminator virus.  

PubMed

Conditionally replication competent adenoviruses (Ads) that selectively replicate in cancer cells and simultaneously express a therapeutic cytokine, such as melanoma differentiation associated gene- 7/Interleukin-24 (mda-7/IL-24), a Cancer Terminator Virus (CTV-M7), hold potential for treating human cancers. To enhance the efficacy of the CTV-M7, we generated a chimeric Ad.5 and Ad.3 modified fiber bipartite CTV (Ad.5/3-CTV-M7) that can infect tumor cells in a Coxsackie Adenovirus receptor (CAR) independent manner, while retaining high infectivity in cancer cells containing high CAR. Although mda-7/IL-24 displays broad-spectrum anticancer properties, pancreatic ductal adenocarcinoma (PDAC) cells display an intrinsic resistance to mda-7/IL-24-mediated killing due to an mda-7/IL-24 mRNA translational block. However, using a chemoprevention gene therapy (CGT) approach with perillyl alcohol (POH) and a replication incompetent Ad to deliver mda-7/IL-24 (Ad.mda-7) there is enhanced conversion of mda-7/IL-24 mRNA into protein resulting in pancreatic cancer cell death in vitro and in vivo in nude mice containing human PDAC xenografts. This combination synergistically induces mda-7/IL-24-mediated cancer-specific apoptosis by inhibiting anti-apoptotic Bcl-xL and Bcl-2 protein expression and inducing an endoplasmic reticulum (ER) stress response through induction of BiP/GRP-78, which is most evident in chimeric-modified non-replicating Ad.5/3- mda-7- and CTV-M7-infected PDAC cells. Moreover, Ad.5/3-CTV-M7 in combination with POH sensitizes therapy-resistant MIA PaCa-2 cell lines over-expressing either Bcl-2 or Bcl-xL to mda-7/IL-24-mediated apoptosis. Ad.5/3-CTV-M7 plus POH also exerts a significant antitumor 'bystander' effect in vivo suppressing both primary and distant site tumor growth, confirming therapeutic utility of Ad.5/3-CTV-M7 plus POH in PDAC treatment, where all other current treatment strategies in clinical settings show minimal efficacy. PMID:24236457

Sarkar, S; Azab, B; Quinn, B A; Shen, X; Dent, P; Klibanov, A L; Emdad, L; Das, S K; Sarkar, D; Fisher, P B

2014-01-01

164

[Determination of the frequency of human bocavirus and other respiratory viruses among 0-2 years age group children diagnosed as acute bronchiolitis].  

PubMed

Acute bronchiolitis, mostly seen in infants and younger children, is a lower respiratory tract infection frequently caused by viral agents. We aimed to determine the frequency of a broad panel of respiratory viruses including human bocavirus (HBoV) and to assess the clinical characteristics of acute bronchiolitis in a group of children under 24 months of age. A total of 62 children (45 male, 17 female; age range: 0-2 years) with the initial diagnosis of acute bronchiolitis and 33 healthy children (21 male, 12 female; age range: 0-2 years) as control group who were admitted to the Pediatrics Department of Mersin University Hospital, southern Turkey, from January to July 2010 were included in the study. Nasopharyngeal aspirates were collected from the study groups and the detection of respiratory viruses [respiratory syncytial virus (RSV) A & B; rhinovirus (RV); human metapneumovirus (hMPV) A & B; influenza virus type A [H1N1, H3N2, H1N1v], B & C; parainfluenza virus (PIV) type 1, 2, 3 & 4A/B; adenovirus (AdV); HBoV; coronavirus (CoV 229E); enterovirus (EV)], were performed by using a commercial system namely CLART®Pneumovir (Clinical Array Technology, Genomica, Spain) based on the principle of multiplex polymerase chain reaction (M-PCR) and DNA microarray. Demographic features, clinical and laboratory findings of the patients, treatment protocols and the relationship between the length of hospitalization and the viral agents determined were also evaluated. Of the 62 samples collected from bronchiolitis cases, at least one virus was detected in 52 (83.9%) and viral co-infections were detected in 31 (50%) of them. Including the co-infections, RSV was the most commonly identified virus (n= 21; 33.9%), followed by influenza A [H1N1] (n= 18; 29%), RV (n= 18; 29%), hMPV (n= 13; 21%), PIV (n= 10; 16.1%), AdV (n= 5; 8%), HBoV (n= 3; 4.8%) and EV (n= 1; 1.6%). Of the 33 samples from healthy children, at least one virus was detected in 21 (63.6%) and viral co-infections were detected in seven (21.2%) samples. Including the co-infections, the most commonly detected virus was RV (n= 10; 30.3%), followed by influenza A [H1N1] (n= 6; 18.1%), AdV (n= 6; 18.1%), RSV (n= 4; 12.1%) and PIV (n= 3; 9%), however HBoV and hMPV were not detected in the control group. The differences of demographic features (age, gender, history of atopy, exposure of smoking, length of breast-feeding, presence of school-age sibling) and frequency of virus detection (83.9% and 63.6%, respectively) between the patient and control groups were not statistically significant (p> 0.05). The most common complaints of patients on admission were cough (100%), runny nose (82.3%) and respiratory difficulty (71%), whereas fever was present in 21 (33.9%) patients. The most common findings on physical examination were prolonged expirium (98.4%), rhonchi (98.4%), rales (80.6%), tachypnea (71%) and tachycardia (67.7%). Pulmonary graphies revealed that diffuse air trappings were more common in virus-associated bronchiolitis (36/52; 69.2%) cases, on the other hand infiltrations were more common (6/10; 60%) in patients who were virus-negative (p< 0.05). The demographic features, clinical and laboratory findings, clinical severity scores, hospitalization rates and duration of hospitalizations in bronchiolitis cases did not show statistically significant differences between the viral agents (p> 0.05 for each parameter). However the rates of antibiotic and steroid use in hospitalized patients (24/34 and 5/34, respectively) were significantly higher than those of outpatients (7/28 and 0/28, respectively) (p= 0.001 and p= 0.03). Our data indicated a high rate (~84%) of respiratory viruses in children with bronchiolitis in the Mersin province and the detection of hMPV (21%) and HBoV (4.8%) only in the patient group encouraged their roles in the etiology of acute brochiolitis. It was concluded that viral etiology should be investigated in selected cases to prevent unnecessary antibiotic treatment and to initiate appropriate antiviral therapy when necessary. PMID:24819262

Uyar, Mustafa; Kuyucu, Necdet; Tezcan, Seda; Aslan, Gönül; Tasdelen, Bahar

2014-04-01