bluuebunny/crossref_metadata_2025_split huggingface dataset:
Parameters of CpG oligodeoxynucleotide‐induced protection against intravaginal HSV‐2 challenge
Authors: Sajic Dusan , Ashkar Ali A. , Patrick Amy J. , McCluskie Michael J. , Davis Heather L. , Levine Kenneth L. , Holl Richard , Rosenthal Kenneth L. 
Published: October 2003

AbstractSynthetic oligodeoxynucleotides (ODN) containing unmethylated CpG dinucleotides within the context of certain flanking bases (CpG motifs) have been shown to induce potent innate and adaptive immune responses. Vaginal delivery of CpG ODN alone protects mice from vaginal herpes simplex virus type‐2 (HSV‐2) challenge. Here, we investigated the importance of timing of delivery, formulation, route and dose of vaginally administered CpG ODN in the prevention or treatment of intravaginal (IVAG) HSV‐2 infection. Mice treated intravaginally with CpG ODN containing a phosphorothioate backbone 24 hours prior to IVAG HSV‐2 challenge survived infection, showed minimal vaginal pathology, and had virtually no detectable virus in vaginal washes, when compared to mice treated with non‐CpG ODN. Genital treatment of HSV‐2 infected mice with CpG ODN 4 hours after infection resulted in increased survival and decreased pathology and vaginal virus titers, whereas treatment of infected mice with CpG ODN 24 and 72 hours after IVAG HSV‐2 infection had no effect on disease progression. Both liquid and solid (delivered on a bio‐erodible muco‐adhesive film) formulations of CpG ODN were effective in protection against genital HSV‐2 following vaginal delivery. Lastly, IVAG delivery of 10 μg of CpG ODN protected as well as a 100 μg dose. J. Med. Virol. 71:561–568, 2003. © 2003 Wiley‐Liss, Inc.
Virological and biochemical relapse according to YMDD motif mutant type during long‐term lamivudine monotherapy
Authors: Akuta Norio , Suzuki Fumitaka , Kobayashi Mariko , Matsuda Marie , Sato Junko , Takagi Kimiko , Tsubota Akihito , Suzuki Yoshiyuki , Hosaka Tetsuya , Someya Takashi , Kobayashi Masahiro , Saitoh Satoshi , Arase Yasuji , Ikeda Kenji , Kumada Hiromitsu 
Published: October 2003

AbstractWhether the type of lamivudine‐resistant virus in hepatitis B virus (HBV) influences the clinical outcome, it is not completely understood. We evaluated the serial changes in YMDD motif mutant in 60 Japanese genotype C–HBV patients who received long‐term lamivudine monotherapy. YIDD or YVDD alone tended to stop shifting to the mixed type (YVDD and YIDD) within 12 months after the detection of mutant virus. Hence, the characteristics, virological relapse (DNA breakthrough) and biochemical relapse (breakthrough hepatitis) of 49 patients, who could be classified into three types (continuous YVDD, continuous YIDD, and the mixed type), were investigated. YVDD and YIDD type tended to have the opposite background with regard to age, histology, and viral load. The mixed and YIDD types tended to have similar backgrounds, except for viral load. In the mixed type, both the HBeAg‐positive rate and viral load as risk factors for emergence of the mutant tended to be high. Mutant virus, DNA breakthrough and breakthrough hepatitis emerged significantly earlier in the mixed type than the two other types. The incidence of severe breakthrough hepatitis accompanied by icteric flare‐up tended to be higher in the mixed type than the other types. Our results suggest that the YMDD motif mutant type might emerge from different backgrounds and modulate the virological and biochemical relapse after the emergence. Large‐scale studies of each mutant type should be conducted in the future to confirm these findings. J. Med. Virol. 71:504–510, 2003. © 2003 Wiley‐Liss, Inc.
Behavior of the dengue virus in solution
Authors: Sithisarn Patchima , Suksanpaisan Lukkana , Thepparit Chutima , Smith Duncan R. 
Published: October 2003

AbstractThe dengue virus consists of four antigenically related but distinct viruses, termed Dengue virus 1–4 (DEN 1–4). We have established that the dengue virus loses infectivity over time in solution in an exponentially declining manner. The four strains examined (one from each serotype) have half‐lives that range from 2.5 to 7.5 hr in defined medium. The half‐life is temperature and pH‐dependent and is affected by the nature of the host cell in which it is produced, but is not dependent upon the presence of either Mg2+ ions or chelating agents. Electron microscopy (EM) of solutions of the dengue virus show almost complete virus aggregation after 24 hr at room temperature, while RT‐PCR shows an intact RNA genome. These results show that the solution environment of the dengue virus is an important determinant of dengue virus infectivity. J. Med. Virol. 71:532–539, 2003. © 2003 Wiley‐Liss, Inc.
Broadly cross‐reactive mimotope of hypervariable region 1 of hepatitis C virus derived from DNA shuffling and screened by phage display library
Authors: Zhang Xin‐Xin , Deng Qiang , Zhang Shen‐Ying , Liu Jing , Cai Qian , Lu Zhi‐Meng , Wang Yuan 
Published: October 2003

AbstractThe hypervariable region 1 (HVR1) is the target of neutralizing antibodies but with isolate specificity. The aim of this study was to obtain immunogenic mimotopes of HVR1, which can react broadly with different HVR1 antibodies and could be one of the candidate immunogens in an effective vaccine against HCV. Thirty‐one HVR1 cDNA fragments were digested by DNase I into a pool of random fragments and reassembled by repeated cycles of annealing in the presence of DNA polymerase to their original size. The shuffled HVR1 was then inserted into the gene III phagemid vector pCANTAB‐5E and displayed on the surface of the phage. Eight individual phages were selected after four rounds of biopanning against anti‐HVR1. ELISA was carried out on immobilized purified phages, respectively, to detect their reactivity with a panel of sera. DNA sequences of the inserts were analyzed and compared with the consensus sequences defined by Puntoriero et al. [(1998) EMBO J 17:3521–3533]. The reactivity of the eight selected clones to the 30 sera were from 53.3 to 80%. Among these, phage 13 (ETYVSGGSAARNAYGLTSLFTVGPAQK, aa 384–410) reacted most broadly. None of the selected sequences encoded for peptides corresponded to known HVR1 from original viral isolates. The two high reactive phages had the similar amino acid sequences with the consensus, which might play a particular role in determining the frequency of reactivity. In conclusion, this study has used effectively DNA shuffling combined with phage display technology to identify broadly cross‐reactive mimotopes recognized by human polyclonal antibodies. Mimotope 13 and 23 appeared to be most reactive immunologically and could be candidate immunogens. Efforts are now underway to identify their neutralizing antibodies by immunization of animals. J. Med. Virol. 71:511–517, 2003. © 2003 Wiley‐Liss, Inc.
Mathematical model to simulate the cellular dynamics of infection with human herpesvirus‐6 in EBV‐negative infectious mononucleosis
Authors: Wang Guanyu , Krueger Gerhard R.F. , Buja L. Maximilian 
Published: October 2003

AbstractAcute infection with human herpesvirus‐6 induces physiological cell proliferation in persons without major immune deficiency. It thus can serve as a parameter to validate a mathematical model designed to simulate cell proliferation under physiological and pathological conditions. Such a mathematical model is presented to simulate various cell changes of the T‐cell immune system during the course of HHV‐6 infection. Model development follows several steps, beginning with a basic model containing physiological T‐cell pools to the introduction of infectious stimuli in the final model. A search algorithm designed to optimize the system parameters, as well as initial variables of the model, is presented. The results of simulation runs for acute HHV‐6 infection of the final computational model correspond well to the data, as documented in human patients; they suggest that the computational model presented for the simulation of T‐cell levels in a given viral infection may well serve as a tool for similar studies of other viral infections, including those that lead to cellular aplasia or neoplasia. J. Med. Virol. 71:569–577, 2003. © 2003 Wiley‐Liss, Inc.
Evaluation of antibody‐based and nucleic acid‐based assays for diagnosis of hepatitis E virus infection in a rhesus monkey model
Authors: Zhang Jun , Ge Sheng X. , Huang Guo Y. , Li Shao W. , He Zhi Q. , Wang Ying B. , Zheng Ying J. , Gu Ying , Ng Mun H. , Xia Ning S. 
Published: October 2003

AbstractWe have evaluated four hepatitis E virus (HEV) specific antibody assays, using sequential samples taken from 86 rhesus monkeys at intervals for up to 86 weeks after they had been infected with different doses of HEV. The animals are a common experimental model of hepatitis E. The large collection of sequential samples used avoids uncertainties encountered in previous studies regarding the precise infection status of study subjects and minimizes bias due to the individuality of response to infection. One assay (YES IgG) was produced with synthetic peptides; the others (E2 IgM, E2 IgG, and GL IgG) were produced with recombinant antigens. The results were compared with the viral RNA contents of the serum and stool samples and the occurrence of these virological and immunological markers in the course of the infection was temporally related to the development of hepatitis. Diagnostic utility of the markers was assessed according to their response rates and prevalence at different times in the course of infection. All the animals produced E2 IgG and developed viremia and all but one also produced E2 IgM and excreted the virus in stool, whereas response rates for the other antibodies were lower and decreased with virus dose. Hepatitis occurred over a period of 4 weeks between 3 and 7 weeks after infection. Virological activity occurred mainly during the incubation period and the prevalence of viral markers declined rapidly after the onset of hepatitis. Production of the E2 antibodies immediately preceded the onset of hepatitis, and this was followed about one week later by production of the other antibodies. Seroprevalence E2 IgM reached a peak value 3 weeks after the onset of hepatitis, whereas seroprevalence of GL IgG and YES IgG peaked after the disease had subsided. E2 IgG persisted in all animals for the entire duration of the experiment of up to 86 weeks and possibly beyond and, thus, can serve as a useful epidemiological marker of HEV infection. J. Med. Virol. 71:518–526, 2003. © 2003 Wiley‐Liss, Inc.
Effector HIV‐specific cytotoxic T‐lymphocyte activity in long‐term nonprogressors: Associations with viral replication and progression
Authors: Keoshkerian Elizabeth , Ashton Lesley J. , Smith Danielle G. , Ziegler John B. , Kaldor John M. , Cooper David A. , Stewart Graeme J. , Ffrench Rosemary A. 
Published: October 2003

AbstractEx vivo effector cytotoxic T‐lymphocyte (CTL) activity was assessed in 27 members of the Australian Long‐Term Nonprogressor cohort and correlated with genetic, virological, and immunological markers. The 27 individuals were antiretroviral naive with CD4+ T‐cell counts of >500 cells/μl for more than 8 years after human immunodeficiency virus type 1 (HIV‐1) infection. Effector CTL activity was determined using a standard ex vivo chromium release assay. Individuals with CTL activity (HIV‐1 envIIIB or pol or gag) were then compared to those without CTL activity in relation to plasma HIV‐1 RNA, ICD p24 antigen, β2‐microglobulin, CD4 and CD8 T‐cell counts, CCR5 and CCR2b genotypes, and progression to CD4 <500 cells/μl or commencement of antiretroviral treatment. Of the 27 individuals examined, 19 had no detectable effector CTL activity. The eight individuals with detectable CTL activity had significantly higher plasma levels of HIV‐1 RNA (P = 0.014), immune complex dissociated p24 antigen (P = 0.006), and β2‐microglobulin (P = 0.009). There was increased risk of progression within 4 years of study entry in individuals with detectable effector CTL activity, higher plasma levels of HIV‐1 RNA, higher β2‐microglobulin levels, and higher immune complex dissociated p24 antigen levels at enrollment (P = 0.017, P = 0.004, P = 0.027, P = 0.008 respectively). Multivariate analysis demonstrated viral load remained the strongest predictor of disease progression within this group (P = 0.017). There were no significant associations between CTL response and chemokine receptor genotype. These findings demonstrate the importance of HIV replication in generating an effector CTL response and show that effector CTL activity may be an early predictor of progression in people with long‐term asymptomatic HIV infection. J. Med. Virol. 71:483–491, 2003. © 2003 Wiley‐Liss, Inc.
Moderate variation of the oncogenic potential among high‐risk human papillomavirus types in gynecologic patients with cervical abnormalities
Authors: Szőke Krisztina , Sápy Tamás , Krasznai Zoárd , Hernádi Zoltán , Szládek Györgyi , Veress György , Dillner Joakim , Gergely Lajos , Kónya József 
Published: October 2003

AbstractThe oncogenic potential of human papillomavirus (HPV) infection was assessed by following the disease course in 455 patients who had had a routine diagnostic Hybrid Capture HPV test due to squamous cell abnormalities of the uterine cervix as detected by cytology and/or colposcopy. At entry, 308 patients had cytologic atypia classified as P3 by the Papanicolau classification, 168 had a positive high‐risk HPV test, and 23 were infected only with low‐risk HPV. The patients were followed‐up using the patient registry until the endpoint of histologically diagnosed cervical intraepithelial neoplasia (CIN). High‐grade CIN was diagnosed in 75 surgical biopsies. High‐risk HPV infection (relative risk: 76.8 CI95: 23.7–249.5), cytologic atypia (RR: 16.2 CI95: 3.9–66.6), and age above 35 (RR: 1.99 CI95: 1.26–3.16) were independent risk factors for high‐grade CIN, while the viral load did not predict oncogenic progression (P = 0.47). After PCR–RFLP typing, the high‐risk types were classified into groups as follows: (1) types 16 and 18, (2) types 45, 52, and 56, (3) types 31, 33, 35, 51, and 58. The relative risks of high‐grade CIN were 119.1 (CI95: 36.2–390.9) for group 1, 44.4 (CI95: 9.8–201) for group 2, and 39.7 (CI95: 10.9–144.8) for group 3, respectively. The risk ratios between the groups of high‐risk types were found to differ at most by a factor of 2.98 (corrected P value: 0.007) indicating that the oncogenic potential varies moderately within the high‐risk group of HPVs. J. Med. Virol. 71:585–592, 2003. © 2003 Wiley‐Liss, Inc.
Modifications and substitutions of the RNA extraction module in the ViroSeq™ HIV‐1 genotyping system version 2: Effects on sensitivity and complexity of the assay
Authors: Stürmer Martin , Berger Annemarie , Doerr Hans‐Wilhelm 
Published: October 2003

AbstractGenotypic testing for HIV‐1 resistance to anti‐retroviral drugs has become accepted widely as a routine method to guide anti‐retroviral therapy. However, implementation into routine high‐throughput laboratory diagnosis is difficult due to the complexity of the assay. A commercially available assay is the ViroSeq™ HIV‐1 Genotyping System (Applied Biosystems, Weiterstadt, Germany). We modified and substituted the RNA extraction module to optimize the proportion of samples amplified successfully as follows: 1 ml plasma was concentrated by ultracentrifugation and extracted according to the manufacturer's instructions (Kit), by substituting the lysis buffer (Roche, Roche Diagnostics GmbH, Mannheim, Germany), and by using the QIAamp Viral RNA Kit (Qiagen GmbH, Hilden, Germany) with elution volumes of 60 (Q60) or 50 μl (Q50). Overall Q50 showed a higher success rate (97%) than the other extraction modules used (range 88–91%). In samples with a viral load range of 1,000–4,999 copies/ml, Q50 was superior (95 vs. 65% to 83%), while in samples with a viral load range of 5,000–9,999 copies/ml or those with 10,000 or more copies/ml, the success rate of the extraction procedures showed no significant differences. In 18 samples, which were negative using the Kit or Roche extraction, Q60 resulted in 7/18 positive results; in addition the Q50 was successful in amplifying 7/10 of the Q60 negative samples. When investigating samples with a measurable viral load of less than 1,000 copies/ml or lower, Q50 had the highest success rate with 80% compared to the other procedures (33–63%). A statistically significant new cut‐off could be defined for Q50 at a value of 250 copies/ml. The results showed clearly that the ViroSeq System is suitable for analyzing the HIV‐1 genotype over a wide range of viral loads but could be improved significantly when substituting the RNA extraction module with Q50 without using a nested PCR protocol. This is of great importance as it avoids further time‐ and cost‐intensive steps. J. Med. Virol. 71:475–479, 2003. © 2003 Wiley‐Liss, Inc.
Buccal exposure to human papillomavirus type 16 is a common yet transitory event of childhood
Authors: Mant Christine , Kell Barbara , Rice Philip , Best Jennifer M. , Bible Jon M. , Cason John 
Published: October 2003

AbstractHigh‐risk human papillomaviruses, such as type 16 (HPV‐16), are established etiological agents for cervical carcinoma. In most cases, this virus is transmitted sexually, though can also be spread from mother to infant at delivery. We have demonstrated previously a high prevalence (∼52%) of HPV‐16 DNA in the mouths of prepubertal children, albeit with low levels of transcription [Rice et al., 2000]. We investigated whether childhood buccal infections with HPV‐16 are persistent or transient and whether children became infected through contact with their immediate family members. Two groups of children were selected: one group were all initially HPV‐16 E5 DNA‐positive in sensitive polymerase chain reaction tests of swabs from their buccal mucosa (n = 20), and the other group consisted of children who were all HPV‐16 E5‐negative (n = 19). Thirty months later, a second oral swab was collected from each child and tested for HPV DNA. At this second visit, 40% of the HPV‐16‐positive group had no detectable HPV‐16 DNA; conversely, 63% of children who were originally HPV‐16‐negative had now acquired the virus. Three months later, a third sample was collected from eight children and their immediate families (seven were HPV‐16 E5 DNA‐positive at the second visit). Amongst the family samples tested, in two families a single previously untested child was HPV‐16 DNA‐positive. It is concluded that HPV‐16 DNA in the oral cavities of children is a transient event and is most probably acquired from their peers. J. Med. Virol. 71:593–598, 2003. © 2003 Wiley‐Liss, Inc.
Molecular epidemiology of Kaposi's sarcoma‐associated herpesvirus/human herpesvirus 8 strains from Russian patients with classic, posttransplant, and AIDS‐associated Kaposi's sarcoma
Authors: Kadyrova Elena , Lacoste Vincent , Duprez Renan , Pozharissky Kazimir , Molochkov Vladimir , Huerre Michel , Gurtsevitch Vladimir , Gessain Antoine 
Published: October 2003

AbstractWe report the molecular characterization of 38 new Kaposi's sarcoma‐associated herpesvirus (KSHV) strains from Russian patients with either classic (25 cases), epidemic/AIDS‐associated (7 cases), or posttransplant/immunosuppressed patients (6 cases), or Kaposi's sarcoma (KS). While a complete sequence of the K1 gene (870 bp) was obtained from 30 strains, only partial sequences of the hypervariable regions VR1 (372 bp) and/or VR2 (381 bp) of the K1 gene were obtained from eight strains of KS paraffin blocks. Sequence comparison and phylogenetic studies indicate that the novel KSHV strains belong to either the A subtype (28 cases) or the C subtype (10 cases). Within the 28 strains of A subtype, 24 (86%) belong to the large A′ subgroup, mostly A1 and A1′ clades, and 4 belong to the A″ subgroup, mostly A3 clade. Within the 10 strains of subtype C, 4 were of C′ subgroup, and 6 of the C″. Some molecular variants of subtype A′ were observed, with 3 strains exhibiting an insertion of a single amino acid at the position 65 and 2 strains (both from AIDS‐KS) with an unique deletion of 17 amino acids in the VR2 region. Polymerase chain reaction‐based subtyping of the K14.1 genomic region indicated that most (23/32) of the novel strains belonged to the P subtype. The results indicate that despite a wide genetic diversity of A and C K1 subtypes of KSHV strains present in Russia, most are closely related and belong to the A1 or A1′ molecular clades suggesting a common origin. This study also expands the data regarding the absence of any correlation between a K1 molecular subtype and a specific KS type (classic, epidemic, or posttransplant), as well as between the K1 and K14.1 molecular subtypes. J. Med. Virol. 71:548–556, 2003. © 2003 Wiley‐Liss, Inc.
Detection of enterovirus in human skeletal muscle from patients with chronic inflammatory muscle disease or fibromyalgia and healthy subjects
Authors: Douche‐Aourik Fatima , Berlier Willy , Féasson Léonard , Bourlet Thomas , Harrath Rafik , Omar Shabir , Grattard Florence , Denis Christian , Pozzetto Bruno 
Published: October 2003

AbstractEnterovirus RNA has been found previously in specimens of muscle biopsy from patients with idiopathic dilated cardiomyopathy, chronic inflammatory muscle diseases, and fibromyalgia or chronic fatigue syndrome (fibromyalgia/chronic fatigue syndrome). These results suggest that skeletal muscle may host enteroviral persistent infection. To test this hypothesis, we investigated by reverse transcription‐polymerase chain reaction (RT‐PCR) assay the presence of enterovirus in skeletal muscle of patients with chronic inflammatory muscle diseases or fibromyalgia/chronic fatigue syndrome, and also of healthy subjects. Three of 15 (20%) patients with chronic inflammatory muscle diseases, 4 of 30 (13%) patients with fibromyalgia/chronic fatigue syndrome, and none of 29 healthy subjects was found positive. The presence of VP‐1 enteroviral capsid protein was assessed by an immunostaining technique using the 5‐D8/1 monoclonal antibody; no biopsy muscle from any patient or healthy subject was found positive. The presence of viral RNA in some muscle biopsies from patients exhibiting muscle disease, together with the absence of VP‐1 protein, is in favor of a persistent infection involving defective viral replication. J. Med. Virol. 71:540–547, 2003. © 2003 Wiley‐Liss, Inc.
Frequency of human cytomegalovirus‐specific T cells during pregnancy determined by intracellular cytokine staining
Authors: Fujikawa Tomoko , Numazaki Kei , Asanuma Hideomi , Kudo Ryuichi , Tsutsumi Hiroyuki 
Published: October 2003

AbstractThe characteristics of cytomegalovirus (CMV)‐specific T‐cell immunity was investigated in pregnant women with primary, latent, or reactivated CMV infection, and in a comparative group of non‐pregnant women. Forty‐six pregnant and 8 non‐pregnant women were examined based on the presence of serum antibody activity against CMV and viral excretion in urine. The frequency of CMV‐specific CD4+ T cells in peripheral blood lymphocytes was determined by staining for intracellular cytokines, interferon (IFN)‐γ, and tumor necrosis factor (TNF)‐α. There was no change in the frequencies of CMV‐specific CD4+ T cells in CMV‐seropositive normal non‐pregnant and pregnant women at any gestation. However, the frequency of CMV‐specific CD4+ T cells in pregnant women associated with CMV reactivation or reinfection was significantly higher than in CMV‐seropositive normal pregnant and non‐pregnant women. There were no CMV transmissions to the infants of all these women. These CMV‐specific T cells responses in pregnant women may contribute some to block the intrauterine CMV infection in their infants. J. Med. Virol. 71:527–531, 2003. © 2003 Wiley‐Liss, Inc.