bluuebunny/crossref_metadata_2025_split huggingface dataset:
Rapid detection of virus genome from imported dengue fever and dengue hemorrhagie fever patients by direct polymerase chain reaction
Authors: Morita Kouichi , Maemoto Tatsuo , Honda Shoji , Onishi Kenji , Murata Misako , Tanaka Mariko , Igarashi Akira 
Published: March 2007

AbstractSerum specimens from patients with imported dengue fever and dengue hemorrhagic fever were directly subjected to reverse transcription and polymerase chain reaction (RT‐PCR) without any RNA purification. The amplified virus genome was detected within 3 hours. The results of PCR corresponded closely to the results of virus isolation using cultured mosquito cells, suggesting that direct RT‐PCR procedure greatly facilitates rapid diagnosis of dengue infection. © 1994 Wiley‐Liss, Inc.
Diagnosis of hepatitis C virus‐associated chronic liver disease in India: Comparison of HCV antibody assay with a polymerase chain reaction for the 5′ noncoding region
Authors: Panigrahi Aswini K. , Nanda Santosh K. , Dixit Rajesh K. , Acharya Subrat K. , Zuckerman Arie J. , Panda Subrat K. 
Published: March 2007

AbstractThe relative value of an anti‐hepatitis C virus (HCV) serological assay and reverse transcriptase‐nested polymerase chain reaction assays (RT‐PCR) were investigated for the constant 5′ putative noncoding region of HCV for the diagnosis of HCV‐associated chronic liver diseases in India. One hundred fifteen patients with biopsy proven chronic active hepatitis and 140 cases of cirrhosis of the liver were investigated for anti HCV antibody using a second generation commercial enzyme‐linked immunosorbent assay (ELISA). A proportion of these patients: 42 with chronic hepatitis and 27 with cirrhosis of the liver were analysed further for HCV RNA in the serum using RT‐nested PCR assay. Thirty‐three (12.9%) of the 255 patients were positive for anti‐HCV antibody and 23 of 69 (33.3%) patients were positive for HCV RNA in serum. Fifteen of the 33 (45.5%) anti‐HCV positive patients had HCV RNA in the serum. Eight of 36 (22.2%) HCV seronegative patients tested were found with HCV RNA. This indicates that the diagnosis of HCV infection is not possible if it is based solely on the available serodiagnostic tests. Inclusion of both assays improved the diagnostic efficiency, 18.8% (13/69) were negative for all virological markers associated with HBV and HCV infection. Since a majority of the chronic liver disease patients (143/255 [56%]) were seronegative for either HBV or HCV infection, it is significant that HCV RNA was detected in 38% (8/21) of a randomly selected group from these patients. The antibody assay and PCR were compared using interclass correlation (kappa statistics). Both these tests were found to be independent of each other. Considering the wide strain variation throughout the world, it is suggested that the current commercial anti‐HCV testing is inadequate for the diagnosis of all HCV infections in all geographical regions.
Detection of adeno‐associated virus DNA in human genital tissue and in material from spontaneous abortion
Authors: Tobiasch Edda , Rabreau Michèle , Geletneky Karsten , Laruë‐Charlus Serge , Severin Fabienne , Becker Nikolaus , Schlehofer Jörg R. 
Published: March 2007

AbstractThe human helper virus‐dependent parvovirus, adeno‐associated virus (AAV) has never been associated with disease in humans [Berns et al. (1987): Advances in Virus Research 32:243–306; Siegl et al. (1985): Intervirology 23:61–73]. However, in pregnant mice, infection with AAV induces early abortion [Botquin et al. (1993): Journal of Cancer Research and Clinical Oncology 119:24]. We investigated whether this common human virus may be found in human genital tissue or in curettage material from spontaneous abortion. Using the polymerase chain reaction (PCR) AAV type 2 DNA was amplified in histological sections of 19 of 30 biopsies of the uterine mucosa. In addition, AAV‐2 DNA was detected in abortion material during the first trimester of pregnancy (12/30 cases were positive) but not in material of abortion from the second or third trimester (9 cases). Whereas in tissues from the uterus AAV DNA was found only by PCR, large amounts of viral DNA were detectable by Southern blot analysis in abortion material. In situ hybridization revealed DNA of AAV to be present in the villous moiety (trophoblast) of the placenta but not in the embryo or decidua. In the same cells, AAV proteins (including the replication‐associated rep proteins) were detected by immunofluorescence analysis. These results suggest (1) that AAV infects the uterine mucosa (possibly persistently) and (2) that it can replicate in trophoblast cells. This might disturb placenta development and may play a role in early miscarriage. To assess an immunological correlate of AAV infection of cervix uteri lesions and miscarriage, an enzyme‐linked immunosorbent assay (ELISA) was developed for detecting serum IgG‐ and IgM‐class antibodies to AAV‐2. Compared to a control population, a significantly higher prevalence of IgM type antibodies to AAV‐2 was found in women with spontaneous abortion and cervical intraepithelial neoplasia. Since about 80% of the population are seropositive for AAV‐2 the high prevalence of IgM‐class antibodies might be associated with reactivation of latent AAV or to reinfections due to a decline of IgG‐mediated immunity. In view of the presence of AAV‐2 DNA and proteins in placenta tissue, serological tests might be useful to assess further the hypothesis of a possible role of AAV infection in spontaneous abortion.
Degradation of p53 only is not sufficient for the growth stimulatory effect of human papillomavirus 16 E6 oncoprotein in human embryonic fibroblasts
Authors: Ishiwatari Haruko , Hayasaka Naoto , Inoue Hirokazu , Yutsudo Masuo , Hakura Akira 
Published: March 2007

AbstractCertain types of human papillomavirus (HPV), such as types 16 and 18, are thought to be responsible for the development of cervical carcinomas. The E6 and E7 genes of these viruses have transforming activities in various cultured cells and their mRNAs and proteins are expressed in almost all cervical carcinoma cells. Inactivation of the tumor suppressor p53 protein by the E6 gene is believed to be critical for transformation by these oncogenic HPVs. To determine whether degradation of the p53 protein is, in fact, sufficient for cellular transformation by the E6 gene, the E6 gene of HPV16 was introduced into human embryonic fibroblasts (HEF) using recombinant murine retrovirus and examined whether reduction of the p53 protein could substitute for the E6 function. It was found that HEF cells transfected with the E6 gene showed an increased saturation density and degraded the p53 protein. However, when expression of the p53 protein in normal HEF cells was suppressed by the antisense oligonucleotide of the p53 gene, growth stimulation was not observed. These results show that the E6 gene stimulates growth of HEF cells, but that this activity involves some other E6 gene‐mediated functions than degradation of the p53 protein. © 1994 Wiley‐Liss, Inc.
Simple procedure for discrimination and typing of enteric adenoviruses after detection by polymerase chain reaction
Authors: Allard Annika , Kajon Adriana , Wadell Göran 
Published: March 2007

AbstractA procedure was developed for specific discrimination and typing of the enteric human adenoviruses, Ad40 and Ad41, after polymerase chain reaction (PCR) amplification of a sequence in the hexon gene highly conserved among all 47 sero‐types recognised. By Taq I restriction of the 300 bp amplimers, subgenus F DNA could be discriminated from DNAs of adenoviruses belonging to all other subgenera. Discrimination between Ad40 and Ad41 was subsequently achieved by cleavage with either Cfo I, HinP I, Mae III, Mvn I, and/or Rsa I. Thus, PCR detection of viral DNA combined with restriction analysis of amplified products provides a valuable tool for use in epidemiological studies of diarrhoea of adenoviral aetiology. © 1994 Wiley‐Liss, inc.
Activation of a varicella‐zoster virus‐specific IgA response during acute Epstein‐Barr virus infection
Authors: Karner Wolfgang , Bauer Georg 
Published: March 2007

AbstractThe influence of an acute Epstein‐Barr virus (EBV) infection on the serological parameters of persistent varicella‐zoster virus (VZV) and herpes simplex virus (HSV) was studied. Sera from 161 patients with infectious mononucleosis caused by EBV and 178 age‐matched controls were tested for HSV‐ and VZV‐specific IgA. 98.7 percent of VZV‐IgG‐positive controls were negative for VZV‐IgA, pointing to the stringent control of latent VZV in healthy individuals. During acute EBV infection, 33.8% of VZV‐IgG‐positive infectious mononucleosis patients produced VZV‐specific IgA. This result may be explained either by reactivation of VZV due to transient suppression of cellular immune functions during acute EBV infection or by polyclonal stimulation caused by EBV. Due to the high incidence of HSV‐IgA in healthy HSV‐IgG‐positive individuals, only a marginal effect of acute EBV infection on the appearance of HSV‐specific IgA was found. © 1994 Wiley‐Liss, Inc.
Brain and kidney of progressive multifocal leukoencephalopathy patients contain identical rearrangements of the JC virus promoter/enhancer
Authors: Ault Grace S. , Stoner Gerald L. 
Published: March 2007

AbstractThe kidneys of six progressive multifocal leukoencephalopathy (PML) patients were examined by PCR amplification for the presence of JC virus. Amplification of three different areas of the viral genome from multiple samples of each kidney revealed three that were positive for the virus. The use of a PCR‐based typing assay on all tissue samples, and cloned sequences from the viral coding region from each positive kidney showed that the same viral genome was present in the kidney as in the brain of the patient. Regulatory region clones all had the archetypal promoter/enhancer structure. However, when PCR fragments from the regulatory region were digested with a restriction enzyme which cuts in region D, the region most often deleted in PML‐type promoters, a low level of undigested DNA remained. This DNA refractory to digestion had a rearranged sequence identical to that of the unique rearranged promoter in the brain of each patient. © 1994 Wiley‐Liss, Inc.
Serum hepatitis C virus RNA level as a predictor of subsequent response to interferon‐α therapy in Japanese patients with chronic hepatitis C
Authors: Onto Etsuro , Mizokami Masashi , Nakano Tatsunori , Terashima Hisahiro , Nojiri Osamu , Sakakibara Kenji , Mizuno Makoto , Ogino Masataka , Nakamura Makoto , Matsumoto Yukoh , Miyata Ken‐Ichi , Lau Johnson Y. N. 
Published: March 2007

AbstractSerum hepatitis C virus (HCV) RNA level has been shown to be a good predictor of subsequent response to interferon‐α (IFN) therapy in US patients in whom genotype 1a/1b are both predominant. To determine whether serum HCV RNA level is a predictor of subsequent response to IFN in Japanese patients or not, appropriately collected pre‐IFN therapy serum samples from 35 Japanese patients with chronic HCV infection were studied. Serum HCV RNA level and HCV genotype were determined and correlated with the subsequent response to IFN. Response to IFN was defined by serum alanine transaminase level: complete and sustained response (n = 15), complete response followed by relapse (n = 10), and no response (n = 10). Patients with complete and sustained response had lower pre‐IFN serum HCV RNA level (median: RT‐PCR+, bDNA‐) compared to the complete response to relapse group (median: 5.25 × 106 genome equivalent/ml [eq/ml], P < 0.001) and the no response group (median: 10.63 × 106 eq/ml, P < 0.001). Seven (46.7%) of the complete and sustained response patients had HCV genotype 2a and three patients had a mixture of genotypes 1b and 2a. In contrast, all 10 patients in the complete response to relapse group had genotype 1b whereas 8 of 10 patients in the non‐response group had genotype 1b and 2 had genotype 2b. The patients with HCV genotype 2a had lower serum HCV RNA level than those with 1b (P = 0.002). When the HCV viremia were controlled by stratifying them into < and > 106 eq/ml, patients with genotype 1 had a similar complete and sustained response rate compared to genotype 2. These data indicated that pre‐IFN serum HCV RNA is also a good predictor of subsequent complete and sustained response in Japanese patients with chronic HCV infection. © 1994 Wiley‐Liss, Inc.
GACPAT HIV 1+2: A simple, inexpensive assay to screen for, and discriminate between, anti‐HIV 1 and anti‐HIV 2
Authors: Parry John V. , Connell Jeffrey A. , Reinbott Paul , Mortimer Philip P. , Garcia Ana B. , Avillez Francisca 
Published: March 2007

AbstractA simple and cheap assay suitable for screening for anti‐HIV 1 and anti‐HIV 2 and discriminating between them was evaluated. In it specimens are incubated in U‐bottomed microplate wells coated with anti‐human IgG for 30 min at room temperature. After washing, 100 μl of a 1 in 50 dilution of HIV 1‐coated gelatin particles (Serodia‐HIV 1/2, Fujirebio) are added. Settling patterns are read on the second day: A positive reaction is indicated by adherence of the particles and a negative by a button. The HIV 1 particles are then washed away and HIV 2 particles added. Anti‐HIV 2 reaction patterns are read on the third day. To assess the performance of the modified “GACPAT HIV 1+2” assay a panel of 1,621 serum/plasma specimens was used. It comprised validated anti‐HIV 1 positive (n = 220), anti‐HIV 2 positive (n = 214), dual anti‐HIV 1/anti‐HIV 2 positive (n = 11), and anti‐HIV negative (n = 1,176) serum/plasma specimens.All 434 specimens that contained anti‐HIV 1 or anti‐HIV 2 reacted positively with the homologous particles. The 11 dually positive specimens reacted positively with both HIV 1 and HIV 2 particles. Five (2.3%) anti‐HIV 1 and five (2.3%) anti‐HIV 2 positive specimens gave positive reactions with both particle types, but none of the five cross‐reactive anti‐HIV 2 specimens were dually reactive when the order of particle addition was reversed. One repeat false positive reaction was recorded with the HIV 1 particles and none with the HIV 2 particles, giving specificities of 99.9% and 100%, respectively. Type specificity was 98.8% after the results of the reverse assay had been taken into account. The assay was sensitive for anti‐HIV 1 in seroconversion series and for anti‐HIV 2 in highly diluted specimens.GACPAT HIV 1 +2 is an accurate anti‐HIV assay applicable to serum/plasma. With few exceptions anti‐HIV 1 positive specimens are reactive only on the second day and anti‐HIV 2 positive specimens only on the third day. It thus discriminates anti‐HIV 2 from anti‐HIV 1, probably through depletion of cross‐reacting antibodies by the reaction with the particles initially added. © 1995 Wiley‐Liss, Inc.
Is the detection of anti‐hepatitis C virus core IgM influenced by the presence of serum rheumatoid factor?
Authors: Pawlotsky Jean‐Michel , Remire Jocelyne , Darthuy Françoise , Intrator Liliane , Udin Laure , Dhumeaux Daniel , Duval Jean 
Published: March 2007

AbstractRheumatoid factor (RF) induces false‐positive results in the detection of serum antibodies, especially of the IgM type. About 70% of the patients with chronic hepatitis C have abnormal levels of serum RF. The aim of this study was to determine whether the presence of serum RF could influence the detection of anti‐HCV core IgM, using an assay designed not to pick up RFs by the addition of goat antibodies directed against human IgG in the sample diluent. Serum anti‐HCV core IgM antibodies and RF were sought in 60 patients with chronic hepatitis C. Serum anti‐HCV IgG antibodies and anti‐HCV core IgM antibodies were also sought in 101 patients with high levels of RF. Anti‐HCV core IgM antibodies were found in 45% and serum RF in 72% of the patients with chronic hepatitis C. Neither the prevalence nor the levels of RF differed significantly between IgM positive and negative patients. Eight percent of the 101 patients with raised RF had anti‐HCV antibodies and two of them had anti‐HCV core IgM antibodies. No patient without anti‐HCV antibodies had anti‐HCV core IgM antibodies. These results show that: a) the detection of anti‐HCV core IgM in patients with chronic hepatitis C is independent of the presence of serum RF; b) high titers of serum RF are not responsible for false‐positive results of anti‐HCV IgM tests. The study suggests that the test used could be a confident tool for studies on the significance of anti‐HCV core IgM antibodies in chronic hepatitis C. © 1995 wiley‐Liss, inc.
Infectious virus titer, replicative and syncytium‐inducing capacity of human immunodeficiency virus type 1
Authors: Nkengasong John N. , Peeters Martine , Nys Patrick , Willems Betty , Piot Peter , van der Groen Guido 
Published: March 2007

AbstractThe relationship was investigated between a viral infectious titer in peripheral blood mononu‐clear cells (PBMC) and plasma on the replicative and syncytium‐inducing capacity of human immunodeficiency virus type 1 (HIV‐1) isolates. The replicative capacity was defined as the minimum time required for p24 antigen to become positive in PBMCs or plasma of HIV‐1 infected individuals, cocultured with PBMCs of healthy donors. Syncytium induction was determined by the MT‐2 cell assay and defined as the presence of giant multinucleated cells. The replicative capacity of HIV‐1 in PBMCs of healthy donors correlated with the infectious viral titer in PBMCs, but not in the plasma of HIV‐1 positive patients. Syn‐cytia formation in MT‐2 cells was not related to the infectious viral titer in PBMCs or plasma of HIV‐1 positive patients. These findings suggest that syncytium formation, not replicative capacity, is an intrinsic HIV‐1 phenotype. © 1995 Wiley‐Liss, Inc.
Hepatitis C virus RNA in plasma and blood mononuclear cells in patients with chronic hepatitis C treated with alpha‐interferon
Authors: Ounanian A. , Seigneurin J. M. , Gueddah N. , Rolachon A. , Thelu M. A. , Zarski J. P. 
Published: March 2007

AbstractHepatitis C virus (HCV), a positive stranded RNA virus, is the main causative agent of post‐transfusion and sporadic non‐A non‐B hepatitis worldwide. Paired samples of plasma and peripheral blood mononuclear cells (PBMC) from 11 patients with chronic hepatitis C treated with α‐interferon (IFN) were tested, using a single step polymerase chain reaction (PCR), for the presence of HCV RNA. Before treatment, the viral genome was detected in all the plasma samples and 81.8% of PBMC. After 3 months of treatment, HCV RNA was still present in 63.6% of plasma samples but in only 27.3% of PBMC. A good correlation was observed between serum alanine aminotransferase level normalisation and disappearance of the viral genome in plasma. Among the six responder patients, five relapsed shortly after IFN withdrawal; HCV RNA became detectable again, especially in PBMC. These results show the presence of HCV in PBMC from most patients infected chronically. IFN therapy had an inhibitory effect on viral replication in lymphoid cells, but frequent relapses observed after cessation of treatment with IFN suggested persistence Of HCV in these Cells. © 1995 Wiley‐Liss, Inc.
Comparison of the kinetics of puumala virus specific IgM and IgG antibody responses in nephropathia epidemica as measured by a recombinant antigen‐based enzyme‐linked immunosorbent assay and an immunofluorescence test
Authors: Elgh Fredrik , Wadell Göran , Juto Per 
Published: March 2007

AbstractImmunoglobulin M and G (IgM and IgG) responses were followed up to 6 months in patients with nephropathia epidemica (NE) by an enzyme‐linked immunosorbent assay (ELISA) using a recombinant Puumala virus (PUU) nucleocapsid protein as antigen and an immunofluorescence test (IF) using PUU infected, acetone‐treated cells as antigen. The recombinant protein was produced by cloning and expressing the nucleocapsid encoding gene of PUU as a polyhistidine fusion protein in Escherichia coli. The product was purified over a metal chelating ion affinity column. On admission, all 17 patients had an IgM response by both methods. The IgM titers decreased significantly by both methods 3 months after onset (ELISA P<0.05 and IF P<0.05). Four of six still had detectable IgM, however at low levels, after 6 months. Presence of specific IgG differed significantly on admission between the two methods: by ELISA 8 of 17 had detectable specific IgG, whereas by IF 15 of 17 had specific IgG (P < 0.02). There were 10 significant titer rises between acute and convalescent serum samples in the same patients by both methods. It is concluded that the IgG antibody response differs in the early phase of NE as measured by a method using a recombinant PUU nucleocapsid protein and a method using PUU infected acetone‐treated cells as antigens. Furthermore, the results suggest that it is of importance to rely on specific IgM for serodiagnosis of NE during the acute phase. © 1995 Wiley‐Liss, inc.
Clinical characteristics and antibody profiles of chronic hepatitis C patients: Relation to hepatitis C virus genotypes
Authors: Yuki Nobukazu , Hayashi Norio , Mita Eiji , Hagiwara Hideki , Oshita Masahide , Ohkawa Kazuyoshi , Katayama Kazuhiro , Kasahara Akinori , Fusamoto Hideyuki , Kamada Takenobu 
Published: March 2007

AbstractGenotyping of 179 consecutive Japanese chronic hepatitis C patients was carried out based on the variation in the hepatitis C virus (HCV) core gene. The results were correlated with clinical features and antibody responses toward specific HCV proteins deduced from the nucleotide sequence of genotype I/1 a. Genotypes II/1 b, III/2a, and IV/2b were identified in 138 (77%), 24 (13%), and 12 (7%) patients, respectively. Five patients had double infections. Genotype dependence was observed only for antibody response toward the NS4 (5–1–1) protein, which was infrequent in genotype III/2a patients (33%) compared with genotype II/1 b (81%; P < 0.01) and genotype IV/2b (75%; P < 0.05). Following interferon‐α therapy, sustained aminotransferase normalisation was achieved by 89% (eight of nine) patients without antibody to the 5–1–1 protein and 33% (17 of 51) with it (P < 0.01). These findings indicate that absence of antibody response to the 5–1–1 protein is frequent in genotype III/2a HCV carriers and may serve to predict responses to interferon therapy. © 1995 Wiley‐Liss, Inc.
Renal failure due to BK virus infection in an immunodeficient child
Authors: De Silva L. M. , Bale P. , De Courcy J. , Brown D. , Knowles W. 
Published: March 2007

AbstractWe report a child with cartilage‐hair hypoplasia and Hodgkin's disease who developed progressive renal failure and died following infection with a polyomavirus, BK virus. Renal biopsy showed interstitial inflammation, tubular atrophy, and intranuclear inclusions in tubular epithelium, with large numbers of papovavirus particles within the cells. BK virus infection was confirmed by polymerase chain reaction on renal biopsy material and in urine and the demonstration of a high titre of haemagglutination inhibition (HI) and IgM antibody to the virus in the patient's serum. This case emphasises the need to study in depth any unusual clinical manifestation in immunocompromised patients to delineate better the full clinical impact of less well‐established pathogens such as BK virus. © 1995 Wiley‐Liss, Inc.