bluuebunny/crossref_metadata_2025_split huggingface dataset:
Generation of fiber‐modified adenovirus vectors containing heterologous peptides in both the HI loop and C terminus of the fiber knob
Authors: Koizumi Naoya , Mizuguchi Hiroyuki , Utoguchi Naoki , Watanabe Yoshiteru , Hayakawa Takao 
Published: April 2003

AbstractBackgroundFiber‐modified adenovirus (Ad) vectors can be effective in overcoming the limitations of conventional Ad vectors, specifically their inefficient gene transfer into cells lacking the primary receptor, the coxsackievirus and adenovirus receptor (CAR). Several types of fiber‐modified Ad vectors have been developed. In this study, we evaluated the functionality of several fiber‐modified Ad vectors.MethodsWe developed a simple method based on in vitro ligation to construct Ad vectors containing heterologous foreign peptides in both the HI loop and C terminus of the fiber knob. A functional comparison of Ad vectors containing RGD and/or K7 (KKKKKKK) peptide in the HI loop or C terminus of the fiber knob was performed in several types of human, mouse, and rat cells, including CAR‐positive and ‐negative cells, and tumor cells in mice in vivo.ResultsIn the case of the in vitro experiment, Ad vectors containing RGD peptide in the HI loop of the fiber knob showed a higher level of gene transfer than vectors containing RGD peptide at the C terminus of the fiber knob. Ad vectors containing K7 peptide at the C terminus of the fiber knob showed levels of gene transfer similar to those of Ad vectors containing RGD peptide in the HI loop of the fiber knob, depending on the cell type. Ad vectors containing both peptides in the HI loop or C terminus of the fiber knob showed the highest levels of gene transfer and a broader tropism. For gene transfer into tumor cells in vivo, the Ad vectors containing RGD peptide were the most efficient.ConclusionsIn the experiment using cultured cells, Ad vectors containing both RGD and K7 peptides were the most efficient with a broader tropism. In contrast, in the experiment in vivo, Ad vectors containing RGD peptide in the HI loop of the fiber knob were more efficient than the vectors containing K7 peptide (including double‐modified vectors containing both the RGD and K7 peptides). These comparative analyses could provide a systemic reference for the use of fiber‐modified Ad vectors. Our simple method, in which the peptide of interest can be expressed in Ad vectors in either the HI loop or the C terminus of the fiber knob, or both, could be a powerful tool for gene transfer into mammalian cells in studies of gene function as well as in gene therapy. Copyright © 2002 John Wiley & Sons, Ltd.
Gene electro‐transfer improves transduction by modifying the fate of intramuscular DNA
Authors: Cappelletti Manuela , Zampaglione Immacolata , Rizzuto Gabriella , Ciliberto Gennaro , Monica Nicola La , Fattori Elena 
Published: April 2003

AbstractBackgroundIntramuscular gene delivery through injection of plasmid DNA has long been considered a promising approach for safe and simple in vivo gene expression for vaccination and gene therapy purposes. Recently, intramuscular gene delivery has been improved by applying low‐voltage electric pulses after plasmid injection, a procedure that has been variably called gene electro‐transfer, in vivo electroporation or electrical stimulation. Different types of electrical treatments have been used with excellent results both in terms of transgene expression levels and immunization outcome. This approach, therefore, holds promise for safe gene delivery to animals and humans designed for non‐viral gene therapy and DNA‐based vaccination. The molecular mechanisms underlying this increment in transduction efficiency are, however, still unclear.MethodsPlasmid DNA status and kinetics following gene electro‐transfer was analyzed by different methods (Southern analysis, Q‐PCR and transformation into competent bacteria).ResultsA large amount of plasmid DNA is degraded in the first 4 h post‐injection, with or without electroporation; later, the amount of intramuscular plasmid DNA is higher in electroporated samples. On electroporation, plasmid is partially protected from degradation, presumably by its early compartmentalization into the nuclei of muscle cells.ConclusionsBy investigating the intracellular outcome and persistence of plasmid DNA following simple injection or gene electro‐transfer we provide useful information on the mechanisms of plasmid entry and expression and underline some of the steps that could be taken to further improve this methodology. Copyright © 2002 John Wiley & Sons, Ltd.
Antibody targeted gene transfer to endothelium
Authors: Tan P. H. , Manunta M. , Ardjomand N. , Xue S. A. , Larkin D. F. P. , Haskard D. O. , Taylor K. M. , George A. J. T. 
Published: April 2003

AbstractBackgroundOne of the drawbacks of the currently available vectors for gene therapy is the lack of selectivity in gene delivery. We have therefore investigated a strategy to generate immunoliposomes to target non‐viral vectors to cell surface receptors on endothelium.Materials and methodsWe have developed a novel method of coupling antibodies (Abs) to liposomes complexed to DNA, using mild heat treatment to aggregate the immunoglobulin G (IgG). The interaction of plasmid DNA, liposomes and Abs was measured using a gel retardation assay and a resonant mirror biosensor. The size of the transfection complex was determined by light scattering, and the binding and internalization of the complex to cells was followed using flow cytometry. The transfection ability was tested on cell lines and primary cells in vitro and human corneal or vascular tissues ex vivo.ResultsThe interaction of antibodies with liposomes is relatively stable (t1/2 ≅ 45 min). The size of the liposome, Ab and DNA complex was found to be around 500 nm in 4% BSA. The addition of anti‐transferrin receptor Abs increased the internalization of the liposome‐DNA complex into cells. Abs against both transferrin receptor and E‐selectin were shown to augment transfection efficiency of liposomes to cell expressing the appropriate antigens. They are also shown to be efficient in mediating gene delivery to corneal and vascular tissues ex vivo.ConclusionsWe have shown that our novel vector is capable of in vitro and ex vivo gene delivery to cells and human tissues including cornea, artery and vein. In particular, an Ab against E‐selectin was effective at selectively delivering genes to activated endothelial cells expressing the adhesion molecule. Such a strategy will have applications for targeting these tissues prior to transplantation or autologous grafting, and, in the longer term, may allow in vivo targeting of gene therapy to inflammatory sites. Copyright © 2002 John Wiley & Sons, Ltd.
Generation of dendritic cells from lentiviral vector‐transduced CD34+ cells from HIV+ donors
Authors: Gruber A. , Chen I. , Kuhen K.L. , Wheat J.C. , Law P. , Wong‐Staal F. 
Published: April 2003

AbstractDendritic cells hold promise as adjuvant for immunotherapy for cancer and infectious diseases. We demonstrate that a significant number of cryopreserved peripheral blood CD34+ cells from HIV‐infected subjects can be transduced with a replication‐incompetent lentiviral vector expressing HIV antigens. In addition, untransduced and transduced CD34+ cells from HIV‐infected individuals were able to differentiate into dendritic cells with strong T‐cell stimulatory capacity. Thus, cryopreserved CD34+ cells from HIV‐infected subjects may prove useful for immunotherapy for HIV/AIDS. J. Med. Virol. 70: 183–186, 2003. © 2003 Wiley‐Liss, Inc.
Sequential changes in hepatitis A virus genotype distribution in Estonia during 1994 to 2001
Authors: Tallo Tatjana , Norder Helene , Tefanova Valentina , Ott Kristi , Ustina Valentina , Prukk Tiina , Solomonova Olga , Schmidt Jelena , Zilmer Kai , Priimägi Ludmilla , Krispin Tõnu , Magnius Lars O. 
Published: April 2003

AbstractHepatitis A virus (HAV) isolates from a large outbreak and from non‐outbreak cases in Estonia were characterized by sequencing the aminoterminal VP1 region. From January 1998 to December 1999, a total of 1,084 cases of hepatitis A were reported to the Harjumaa‐Tallinn and Ida‐Virumaa Health Protection Services in Estonia. The attack rate was highest among males aged 15–29. Initial cases were noted to be associated with injecting drug use. IgM anti‐HAV positive sera were available from 107 hospitalized outbreak cases and from 68 patients sampled during 1994 to 2001. HAV RNA was detected in 42% of sera from 1994–1996 and in 88% of sera from 1998–2001. It was possible to obtain HAV sequences from 83 outbreak and 29 background cases. The outbreak strain was represented by five different sequences, all belonging to subtype IIIA. During the outbreak, this IIIA strain also spread into the general population. All available non‐outbreak isolates from 1994 to 2001 but one belonged to genotype IA and formed distinct clusters as compared to isolates from other parts of the world. One subtype IIIA isolate from 1995 was unrelated to the outbreak strain. Subtype IA had been dominating in Estonia during 1994–2001, but the outbreak strain from 1998 to 1999 was IIIA. This subtype was encountered previously in addicts in Sweden during the 1980s and in Norway at the end of the 1990s. This study supports the use of limited sequencing within the aminoterminal VP1 region for studying the molecular epidemiology of hepatitis A. J. Med. Virol. 70: 187–193, 2003. © 2003 Wiley‐Liss, Inc.
Perinatal and intrafamily transmission of hepatitis B virus in three generations of a low‐prevalence population
Authors: Ördög Katalin , Szendrôi Andrea , Szarka Katalin , Kugler Zoltán , Csire Márta , Kapusinszky Beatrix , Xie Jiuru , Csizmadia Katalin , Brojnás Judit , Rusvai Erzsébet , Tempfli Ágnes , Berencsi György 
Published: April 2003

AbstractFamily members of 47 hepatitis B virus (HBV)‐carrier pregnant women were tested for the presence of hepatitis B surface antigen (HBsAg), other markers of HBV infection, and hepatitis A virus (HAV) antibodies. Eleven members of six families were found to be HBV DNA positive. Five of the anti‐HBe‐positive persons were found to be HBV DNA carriers, too. The mean age of the HBV DNA carriers was found to be lower than that of Hbe carriers; therefore, it is suggested that seroconversion to HBe occurs before the resolution of HBV DNA carrier state. Superinfection with hepatitis A virus was not found to influence the elimination of HBV‐carrier state, as there was no correlation found between the hepatitis A exposure and the hepatitis B virus markers in the families. The low HBV prevalence in the population (0.3%) was in contrast to the high prevalence of the families of the HBV‐carrier mothers (27.1%) and family members with HBV markers (50.4%). Significant positive correlation was found in the proportion of HBV‐positive children, and the HBV history of their parents. When fathers were shown to be seronegative, the probability of HBV transmission was reduced by a factor of 6 (12.5% instead of 75%) probably due to reduced viral load and possibly by other factors. Several results indicate, that the noncytocidal hepatitis B virus clearing mechanism suggested by Guidotti et al. [1996, 1999] was effective also in the HBV‐carrier human population. J. Med. Virol. 70: 194–204, 2003. © 2003 Wiley‐Liss, Inc.
Visual gene diagnosis of HBV and HCV based on nanoparticle probe amplification and silver staining enhancement
Authors: Wang Ye‐Fu , Pang Dai‐Wen , Zhang Zhi‐Ling , Zheng Hu‐Zhi , Cao Jun‐Ping , Shen Jun‐Tao 
Published: April 2003

AbstractA visual gene‐detecting technique using nanoparticle‐supported gene probes is described. With the aid of gold nanoparticle‐supported 3′‐end–mercapto‐derivatized oligonucleotide serving as detection probe, and 5′‐end –amino‐derivatized oligonucleotide immobilized on glass surface acting as capturing probe, target DNA was detected visually by sandwich hybridization based on highly sensitive “nano‐amplification” and silver staining. Different genotypes of Hepatitis B and C viruses in the serum samples from infected patients were detected using home‐made HBV, HCV, and HBV/HCV gene chips by the gold/silver nanoparticle staining amplification method. The present visual gene‐detecting technique may avoid limitations with the reported methods, for its high sensitivity, good specificity, simplicity, speed, and cheapness. This technique has potential applications in many fields, especially in multi‐gene detection gene chips coupled with the detection will find applications in clinic. Additionally, resonance Rayleigh light scattering (RLS) spectroscopy is used, for the first time, to judge and monitor the immobilization of gene probes on gold nanoparticle surfaces. J. Med. Virol. 70: 205–211, 2003. © 2003 Wiley‐Liss, Inc.
Diversification of hypervariable region 1 of hepatitis C virus after liver transplantation
Authors: Fan Xiaofeng , Di Bisceglie Adrian M. 
Published: April 2003

AbstractLiver transplantation represents a unique model for studying hepatitis C virus (HCV) evolution. In the current study, we estimated the dynamics of HCV quasispecies evolution over a period of 2.5 years both in sera and in tissues, including three different anatomic sites of the liver in a patient undergoing liver transplantation. The patient displayed an extremely homologous virus population in either serum or liver at each time point after liver transplantation. However, there was a successive diversification of hypervariable region 1 (HVR1) of HCV under such an im‐munocompromised status and the preexistence of matching HVR1 antibodies. This distinct evolutionary pattern of HVR1 suggests that immunosuppressed status does contribute to diminished genetic diversity of HVR1 at a given time point after liver transplantation. However, the dramatic diversification of HVR1 sequence over time is mainly due to the adaptation of new replicative site, the donor liver, indicating that HCV may obtain the higher replication fitness by mutating HVR1 domain. J. Med. Virol. 70: 212–218, 2003. © 2003 Wiley‐Liss, Inc.
Upregulation of endogenous intrahepatic interferon stimulated genes during chronic hepatitis C virus infection
Authors: MacQuillan Gerry C. , Mamotte Cyril , Reed William D. , Jeffrey Gary P. , Allan Jane E. 
Published: April 2003

AbstractThe success of interferon‐α and ribavirin combination therapy for the treatment of chronic hepatitis C viral infection differs between patients. In an attempt to identify predictors of host response to therapy, the levels of mRNA for interferon (IFN) stimulated genes: MxA, PKR, 2′5′ OAS, ISG15, and interleukin 8 (IL‐8), were examined in liver by real‐time RT‐PCR prior to commencement of therapy. The levels of intrahepatic classical IFN stimulated genes, but not IL‐8, in chronic HCV disease (n = 44) were found to be significantly upregulated (P < 0.001) compared to the control cohort (n = 12). The genotype of the infecting HCV strain did not influence IFN stimulated gene expression. These results suggest that the endogenous type 1 IFN antiviral effector pathway is broadly activated during chronic HCV disease, although the levels of mRNA for any of the IFN‐stimulated genes tested did not predict the outcome of combination therapy. J. Med. Virol. 70: 219–227, 2003. © 2003 Wiley‐Liss, Inc.
Rapid and quantitative detection of human adenovirus DNA by real‐time PCR
Authors: Heim Albert , Ebnet Carmen , Harste Gabi , Pring‐Åkerblom Patricia 
Published: April 2003

AbstractRapid diagnosis of human adenovirus (HAdV) infections was achieved by PCR in the recent years. However, conventional PCR has the risk of carry‐over contamination due to open handling with its products, and results are only qualitative. Therefore, a quantitative “real‐time” PCR with consensus primer and probe (dual fluorescence labelled, “TaqMan”) sequences for a conserved region of the hexon gene was designed and evaluated. Real‐time PCR detected all 51 HAdV prototypes. Sensitivity of the assay was ≤15 copies/run and the linear range of quantitation 1.5 × 101 to 1.5 × 108 copies/run. TaqMan PCR gave identical results compared to an established conventional one‐step PCR protocol in 218 (38 positive and 180 negative) of 234 clinical samples including blood, serum, eye swabs, and feces, and had divergent results in 16 samples (15 positive only in TaqMan PCR, all with low copy numbers, and one positive only in conventional PCR), indicating a higher sensitivity of TaqMan PCR. Adenovirus viremia was detected by TaqMan PCR in 4 of 27 (14.8%) paediatric and 8 of 93 (8.6%) adult stem cell transplant recipients but only in 5 of 306 healthy controls (blood donors, 1.6%). Virus loads of pediatric patients (median 1.7 × 105) were significantly higher than in adult patients (median 2.3 × 103) and than in controls (all samples ≤1.7 × 103 copies/ml). A few immunosuppressed children had very high virus loads (up to 1.1 × 1010 copies/ml), which were associated with symptoms of disseminated disease. In conclusion, real‐time PCR is a sensitive and quantitative procedure for the detection of adenovirus infections. J. Med. Virol. 70: 228–239, 2003. © 2003 Wiley‐Liss, Inc.
Single human cytomegalovirus gB genotype shed in multiple sites at the time of diagnosis in renal transplant recipients
Authors: Carraro E. , Granato C.F.H. 
Published: April 2003

AbstractHuman cytomegalovirus (HCMV) is a major cause of morbidity and mortality in immunocompromised patients, such as renal transplant recipients. Analysis of the gene encoding the envelope glycoprotein B (gB) showed that clinical isolates adopted one of the sequence configurations, permitting the isolates to be assigned a gB genotype of 1–4. It has been suggested that HCMV gB genotypes could be correlated with tropism and pathogenesis. A number of reports in the literature refer to shedding of different gB strains, permitting follow‐up of renal transplant recipients. Considering that a single strain might be responsible for the clinical expression of the disease in multiply exposed individuals, the frequency distribution of gB genotypes was examined by nested polymerase chain reaction and restriction fragment length polymorphism in 20 renal transplant recipients at the time of diagnosis. The association between gB genotypes and cellular tropism was determined using blood, saliva, and urine for each patient. HCMV gB genotype 2 was found more frequently than other genotypes (gB2, 40%; gB1, 30%; gB3, 25%; and gB4, 5%) in renal transplant recipients. The gB type did not correlate with tropism for different body sites. All the patients with HCMV infections presumably harbored a single HCMV strain at the time of diagnosis. In multiply exposed patients, the immunomodulation provided by acute HCMV infection could favor later shedding of different strains. J. Med. Virol. 70: 240–243, 2003. © 2003 Wiley‐Liss, Inc.
Contrasting effects of hydroxyurea on cell growth and reduction in Epstein‐Barr virus genomes in EBV‐infected epithelioid cell lines vs. Burkitt's lymphoma cell lines
Authors: Jiang Ru , Kanamori Mikiko , Satoh Yukio , Fukuda Makoto , Ikuta Kazufumi , Murakami Masanao , Sairenji Takeshi 
Published: April 2003

AbstractEliminating Epstein‐Barr virus (EBV) genomes from infected cells is an intriguing theoretical strategy in therapy for EBV‐associated malignant diseases. Respective patterns were characterized for hydroxyurea (HU)‐promoted loss of EBV genomes from EBV‐infected epithelioid cell lines derived from the noncancerous portion of gastric carcinoma tissues and Burkitt's lymphoma (BL) cell lines. Epithelioid cell lines GT38 and PN were less sensitivity to HU than BL cell lines Akata, Raji, and Daudi in terms of cell growth inhibition and cell cycle arrest. On passage in medium with 50 μM HU, the fraction of EBV nuclear antigen (EBNA)‐positive cells was reduced substantially in the BL cell lines, but only slightly in the epithelioid cell lines. EBV DNA was reduced in Akata, Raji, and Daudi cells upon passage in 50 μM HU by 95%, 70%, and 50%, respectively, but by only 10% in GT38 cells, in which EBV DNA reduction was enhanced at increased concentrations of HU. This indicates that EBV genome is more easily lost from BL cell lines than from epithelioid cell lines upon culturing in HU. These findings support the view that the elimination of EBV could be therapeutically effective in EBV‐associated BL by HU. J. Med. Virol. 70: 244–252, 2003. © 2003 Wiley‐Liss, Inc.
Infectious mononucleosis patients temporarily recognize a unique, cross‐reactive epitope of Epstein‐Barr virus nuclear antigen‐1
Authors: McClain Micah T. , Rapp Erin C. , Harley John B. , James Judith A. 
Published: April 2003

AbstractThe spectrum of antibodies against Epstein‐Barr virus nuclear antigen‐1 (EBNA‐1) in patients with a recent history of infectious mononucleosis and nonaffected EBV‐positive individuals has been characterized by epitope mapping. Sera were evaluated for antibodies to all unique maximally overlapping octapeptides of EBNA‐1. Both normal controls and patients with infectious mononucleosis produce IgG antibodies that recognize the glycine‐alanine‐rich portion of EBNA‐1, as previously described. All EBNA‐1 IgG‐positive infectious mononucleosis patients tested, however, consistently produce IgG specific for an additional epitope (aa 398–412 PPPGRRPFFHPVGEA) near the middle of the EBNA‐1 protein. This region was not found to be antigenic in healthy EBV‐seropositive individuals. This region does, however, cross‐react with the sequence PPPGMRPP from the common lupus spliceosomal autoantigen Sm B′ in several infectious mononucleosis patient sera. Patients with recent clinical infectious mononucleosis temporarily recognize a unique cross‐reactive epitope of EBNA‐1 not bound by antibodies from non‐infectious mononucleosis EBV‐positive sera or those with a distant history of IM. J. Med. Virol. 70: 253–257, 2003. © 2003 Wiley‐Liss, Inc.
Infantile viral gastroenteritis: On the way to closing the diagnostic gap
Authors: Simpson R. , Aliyu S. , Iturriza‐Gómara M. , Desselberger U. , Gray J. 
Published: April 2003

AbstractA total of 305 faecal specimens collected from children under the age of 5 who presented with symptoms of acute gastroenteritis either as inpatients at Addenbrooke's Hospital (N = 100) or to General Practitioners in East Anglia (N = 205) during 1999–2001 were tested for the presence of rotavirus, norovirus, sapovirus, enteric adenoviruses (Group F, serotypes 40 and 41), and astrovirus. An aetiologic agent was found in 184 specimens (60.3%). The most commonly found single viral pathogen was rotavirus (27.9%), followed by norovirus (13.4%), enteric adenoviruses (7.9%), astrovirus (2.3%), and sapovirus (1%). Mixed infections were observed in 27 specimens (8.9%), and no aetiologic agent was found in over a third of the specimens tested. These data demonstrate that the diagnostic gap can be reduced considerably through the use of molecular amplification and detection techniques. However, additional work is required to reduce this deficit further by optimising sampling algorithms and by identifying other agents associated with viral gastroenteritis. J. Med. Virol. 70: 258–262, 2003. © 2003 Wiley‐Liss, Inc.