bluuebunny/crossref_metadata_2025 huggingface dataset:
Genotyping of hepatitis C virus isolates by a modified polymerase chain reaction assay using type specific primers: Epidemiological applications
Authors: Widell Anders , Shev Steven , Månsson Siv , Zhang Yong‐Yuan , Foberg Ulla , Norkrans Gunnar , Frydén Aril , Weiland Ola , Kurkus Jan , Nordenfelt Erik
Published: March 2007

A polymerase chain reaction (PCR)‐based assay using primers against the hepatitis C core gene has been described [Okamoto et al. (1992a): Journal of General Virology 73:673–679]. Within the two major HCV genotypes 1 and 2, the Okamoto system identifies two subtypes each (1a,1 b and 2a, 2b, respectively). Typing is achieved by a primary PCR with consensus primers followed by a nested PCR with type specific primers. The original assay was modified by addition of a parallel second PCR identifying the recently described major genotype 3. The assay also identifies in duplicate subtype 1 b (type II by Okamoto), suggested to respond poorly to interferon. Reaction conditions were reviewed and melting temperatures of all typing primers equalised to increase stringency.

The modified system functioned well and typing results were supported by partial core sequencing. The following distribution of genotypes was found in 53 hepatitis C virus (HCV) infected Swedish blood donors: genotype 1a (57%), 3 (19%), 1b (13%), and 2b (11%). In six recipients of HCV infected blood identified in a retrospective study, the recipient HCV genotype was identical to donor HCV genotype. Furthermore, in HCV positive couples identical genotype was observed when only one partner had an external risk factor; whereas genotypes were often diverse if both sex partners had parenteral risk factors. Finally, a cluster of hepatitis C cases in a haemodialysis unit was evaluated retrospectively. Eight patients had genotype 1b, two had mixed 1a and 1b, and one had type 1a.

The modified HCV genotyping assay was of value in examining different epidemiological situations and can be expanded presumably to include future genotypes. © 1994 Wiley‐Liss, inc.
Use of the colonic carcinoma cell line CaCo‐2 for in vivo amplification and detection of enteric viruses
Authors: Pintó Rosa M. , Diez José M. , Bosch Albert
Published: March 2007

The use of the continuous cell line CaCo‐2 as an in vivo amplification system for the detection of fastidious human enteric viruses is reported. CaCo‐2 cells showed an increased sensitivity to laboratory strains of group A rotavirus 3, reovi‐rus3, astrovirus 1, poliovirus 1, coxsackievirus A 24, enterovirus 70, and adenovirus 5, 40 and 41, when compared to a routine host cell line for each virus. Nucleic acids from wild‐type infectious rotavirus, astrovirus, and adenovirus 40 in stool samples of patients with acute gastroenteritis could be amplified after infection of CaCo‐2 cells with trypsin‐pretreated virus inoc‐ula. Virus diagnosis was carried out subsequently by dot‐blot hybridisation with specific cDNA probes. An amplification factor between 10 and 1,000x was obtained by infection of CaCo‐2 cells, thus enabling specific detection of low numbers of a wide range of enteric viruses, and the differentiation between infectious and noninfectious particles. © 1994 Wiley‐Liss, inc.
Mixed cryoglobulinemia type II in chronic hepatitis B associated with HBe‐minus HBV mutant: Cellular immune reactions and response to interferon treatment
Authors: Löhr Hanns , Goergen Bernd , Weber Wolfgang , Godderz Werner , zum Büschenfelde Karl‐Hermann Meyer , Gerken Guido
Published: March 2007

The case of a young female patient with chronic active hepatitis B, vasculitic purpura, edema, and circulating immune complexes due to mixed Cryoglobulinemia is described. Serum transami‐nases were elevated. Serological assays showed hepatitis B surface antigen (HBsAg), antibody to hepatitis B e antigen (anti‐HBe), and antibody to hepatitis B core antigen (anti‐HBc) antibodies but no antibody to hepatitis C virus (anti‐HCV) or antibody to hepatitis delta virus (anti‐HDV) antibodies. Using hepatitis B virus‐polymerase chain reaction (HBV‐PCR) and direct sequencing a precore/core (preC/C) mutant unable to synthesize HBeAg was detected in serum. HBV antigens were demonstrated in the circulating immune complexes. Following 1 month of treatment with interferon‐a 2b 3 miu three times weekly, alanine aminotransferases returned to normal levels while cryoglobulins and immune complexes disappeared from serum. In addition, 2 months after the onset of treatment serum HBV‐DNA was no longer detectable by PCR.

Prior to treatment the analysis of cellular immune reactions of peripheral blood mononu‐clear cells showed a major proliferative response to HBcAg, preS1Ag and HBxAg and a minor response to HBeAg and HBsAg. One month after conclusion of treatment a decline in T‐cell reactivity against all HBV antigens was observed. During clinical response to the therapy, however, a strong proliferative response of T cells to HBcAg and HBeAg was demonstrated.

In conclusion, immune complex disease may complicate chronic hepatitis B in patients expressing HBe‐minus HBV mutants. Treatment with interferon‐a was found to be effective in mixed Cryoglobulinemia even in the presence of HBe‐minus HBV mutants. Antiviral effects of interferon‐a induce a decrease in specific T‐cell recognition of HBV antigens. Clinical response and virus elimination were, however, accompanied by a reconstitution of cellular immune responses to HBV core antigens, i.e., HBcAg and HBeAg. The response to certain T cell epitopes on these antigens may be of pathogenetic relevance for virus elimination. © 1994 Wiley‐Liss, Inc.
Nucleotide sequence of the core region of hepatitis C virus in Pakistan and Bangladesh and the geographic characterisation of hepatitis C virus in South Asia
Authors: Ohno Tomoyoshi , Mizokami Masashi , Tibbs Christopher J. , Nouri‐Aria Kayhan T. , Wu Rong‐Rong , Ohba Ken‐Ichi , Orito Etsuro , Suzuki Kaoru , Mizoguchi Naoto , Nakano Tatsunori , Khan Mobin , Yano Michitani , Kiyosawa Kendo , Williams Roger
Published: March 2007

A large number of complete and partial hepatitis C virus (HCV) sequences have been reported and classified into several genotypes, although none have been reported from South Asia. We have determined and evaluated partial sequences in the core region of HCV obtained from patients with chronic hepatitis in Pakistan and Bangladesh. Nucleotide sequences from these viruses show significant homology with the Japanese HCV‐TR isolate (91.7%–97.9%) and low homology with other Japanese, American, and UK isolates including HCV‐1, HC‐J4, HC‐J6, HC‐J8, and E‐b1 (79.3%–86.2%). The homologies of their deduced amino acids sequence with HCV‐1, HC‐J4, HC‐J6, HC‐J8, E‐b1, and HCV‐TR were 84.3%–89.8%, 85.0–87.9%, 84.1%–86.9%, 84.3%–87.0%, 90.2%–93.1%, and 89.8%–93.5%, respectively. These results suggest that our clones might be classified into the same genotype as HCV‐TR. Further analysis using molecular evolutionary methods strongly supported the classification of these sequences with the HCV‐TR genotype. Moreover, we could not detect any isolates which were closely related to our clones or HCV‐TR in countries outside the South Asian area. These data further support the association of HCV genotypes with distinct geographic regions. © 1994 Wiley‐Liss, Inc.
Elimination of hepatitis delta virus infection after loss of hepatitis B surface antigen in patients with chronic delta hepatitis
Authors: Battegay Manuel , Simpson Lisa H. , Hoofnagle Jay H. , Sallie Richard , di Bisceglie Adrian M.
Published: March 2007

The aim of this study was to evaluate whether patients with chronic hepatitis delta virus (HDV) infection treated with alpha interferon and subsequent loss of hepatitis B surface antigen (HBsAg) eliminate HDV. HDV RNA was detected in 26 of 28 patients with chronic delta hepatitis using the polymerase chain reaction. Seventeen patients in whom HDV RNA was detected were treated with alpha interferon; in 65%, HDV RNA remained detectable during treatment or reappeared after stopping therapy whereas in three patients HDV RNA remained absent (17.5%). HDV RNA became and remained undetectable in serum and liver of two of these three patients who lost HBsAg from serum and in one patient who was intermittently HBsAg negative during therapy. After loss of HBsAg, hepatitis B virus (HBV) DNA was still detectable in the liver, but not HBV RNA, indicating absent or very low HBV replication. Three patients were lost to follow up (17.5%). Two nontreated patients with chronic HDV infection also lost HBsAg during follow up; HDV RNA also became undetectable in their serum. Thus, HDV replication does not persist after the loss of HBsAg. Clearance of HBsAg may be a useful guide to when therapy can be stopped. © 1994 Wiley‐Liss, Inc.
Viral load as a determinant for transmission of human papillomavirus type 16 from mother to child
Authors: Kaye Jeremy N. , Cason John , Pakarian Farzin B. , Jewers Richard J. , Kell Barbara , Bible Jon , Raju Kankipati S. , Best Jennifer M.
Published: March 2007

Whilst genital papillomaviruses are commonly believed to be sexually transmitted, transmission of human papillomavirus type 16 (HPV‐16) from mother to child at delivery has been described previously [Pakarian et al. (in press) British Journal of Obstetrics and Gynaecology]. In order to determine whether viral load in cervical/vaginal cells was an important determinant of transmission 15 pregnant women with HPV‐16 infections were studied. Eight of these women had infants who were positive for HPV‐16 DNA at genital and/or buccal sites. Viral load was estimated by laser densitometry of polymerase chain reaction (PCR) products. The eight mothers—four with a previous history of abnormal smears and two with previous genital warts— who transmitted infection to their infants had significantly higher viral loads (P < 0.05) than those who did not. It is concluded that viral load is an important, but not the sole, determinant for the transmission of HPV‐16 from mother to infant. © 1994 Wiley‐Liss, Inc.
Clinical and immunological investigation of a new combined hepatitis A and hepatitis B vaccine
Authors: Ambrosch F. , Wiedermann G. , André F. E. , Delem A. , Gregor H. , Hofmann H. , D'hondt E. , Kundi M. , Wynen J. , Kunz Ch.
Published: March 2007

As with hepatitis B vaccines, the recently developed hepatitis A vaccine is suitable not only for individual protection, but also for public health control measures. For introduction into routine immunisation programmes, however, hepatitis A vaccine should preferably be combined with other already established vaccines. In particular, a combination of hepatitis A and hepatitis B vaccines would be appropriate. We investigated a new combined hepatitis A/hepatitis B vaccine comparing its tolerability and immunogenicity with that obtained after separate or mixed simultaneous administration of the two components. Three groups of healthy volunteers, each of approximately 50 persons, were included. All were negative for hepatitis A and hepatitis B markers and had normal liver enzyme values. They received hepatitis A (720 ELISA units) and hepatitis B (20 m̈g) vaccines in the deltoid muscle, combined, mixed or separately, according to a 0, 1, 6‐month schedule. Blood samples for determination of antibodies to hepatitis A virus (anti‐HAV) and hepatitis B virus (anti‐HBs) and of serum ala‐nine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were drawn at months 0, 1, 2, 6, and 7. Local and systemic reactions were monitored by means of questionnaires.

The results of our study demonstrate that the combined hepatitis A and B vaccine is well tolerated and highly immunogenic. The seropositivity and seroprotection rates were 100% for both antigens in all groups. Surprisingly, anti‐HAV and anti‐HBs antibody titres after the combined and mixed vaccines were significantly higher compared with the respective monovalent vaccines injected separately. © 1994 Wiley‐Liss, Inc.
Cost‐effectiveness analysis of vaccination against hepatitis A in travellers
Authors: van Doorslaer E. , Tormans G. , van Damme P.
Published: March 2007

Hepatitis A virus (HAV) infection is a substantial risk for travellers from low endemic countries to high endemic destinations. Costs and effects of alternative options for prevention were compared using formal decision analysis. General indications for the optimal prevention of hepatitis A were derived from a cost‐effectiveness analysis. Various possible strategies for prevention of hepatitis A in travellers were compared to doing nothing: active immunisation using either the existing vaccine (HAVRIX 720) or the new vaccine (HAVRIX(tm) 1440); first screening for the presence of HAV antibodies and then vaccinating only susceptibles; and passive immunisation with immunoglobulin. Using a number of assumptions as baseline and for an average duration and frequency of travel from low to high endemic countries, threshold values were obtained for the choice between passive and active immunisation. Passive immunisation remains the most cost‐effective prevention strategy for those expected to travel not more frequently than twice over the next 10 years and for short stays (£7,000–9,000 per infection prevented). For travellers expected to travel three or more times in 10 years or for trips exceeding a period of 6 months, active immunisation before the first trip is the most cost‐effective option (£7,500 or less per infection prevented). When travel frequency increases to once a year in the next 10 years, costs per infection prevented decrease to about £3,500. Screening for the presence of antibodies before vaccination is only justified for older travellers or those leaving from countries with moderate endemicity, i.e., with an average HAV prevalence Of at least 30%. © 1994 Wiley‐Liss, Inc.
Symptomatic mumps virus reinfections
Authors: Gut Jean‐Pierre , Lablache Christel , Behr Sylvie , Kirn André
Published: March 2007

Although natural mumps virus infection is believed to induce lifelong immunity, our laboratory was confronted with 82 patients who developed mumps‐evoking lesions but exhibited serological evidence of a booster immune response, namely a rise or a high titer of virus‐specific IgG, without IgM. In order to provide arguments favoring the existence of recurrent mumps attacks, the age, symptomatology, and humoral response of these patients (group 1) were compared to that of 82 randomly selected true primary infected patients (group 2), 10 parainfluenza virus‐infected patients (group 3), and 20 noninfected mumps‐immune subjects (group 4), Enzyme‐linked immunosorbent assay (ELISA) procedures with different viral antigenic preparations were used for determination of specific IgM, IgA, IgG, IgG subclasses, and IgG avidity. The patients of group 1, older than those of group 2 (28 vs. 10 years, P <0.0001), presented a significantly less severe and less typical symptomatology. Against the whole virus they exhibited IgG of higher avidity (P < 0.001), a lower prevalence and titer of IgA (10 vs. 68%, P < 0.0001 and 278 vs. 5,009, P < 0.001, respectively). Values obtained for IgG 1, 2, and 3 were significantly different between the two groups. Prevalence and absorbance of nucleocapsid‐directed IgG 3 were significantly lower in group 1 (27 vs. 46%, P < 0.01 and 0.444 vs. 0.869, P < 0.01, respectively). A significant discrepancy also allowed patients from group 1 to be distinguished from those of groups 3 and 4. So the presumed mumps‐reinfected patients were different from primary infected ones and presented features reminiscent of a secondary immune response, suggesting the not uncommon occurrence of symptomatic reinfections by the mumps virus. This concept is of importance in medical practice. © 1995 Wiley‐Liss, Inc.
Unbiased usage of T‐cell receptor β variable region genes in peripheral blood cells of hepatitis C patients: No correlation with superantigen effect
Authors: Wang C. C. , Hwang L. H. , Chen D. S. , Yang P. M. , Chiang B. L. , Chen P. J.
Published: March 2007

Hepatitis C virus (HCV) infection frequently causes chronic hepatitis and lack of virus clearance in these patients. In addition, many patients infected by HCV also present with hypergamma‐globulinemia in the early stage of chronic infection. These observations raise a possible viral superantigen effect induced by HCV, because viral superantigen found in human immunodeficiency virus (HIV) or in replication of defective murine leukemia virus (MuLV) is associated with T‐cell dysfunction and polyclonal activation of B cells. The possibility was investigated of whether HCV encodes any superantigen by analyzing the usage of T‐cell receptor (TCR) from the peripheral blood lymphocytes (PEL) of patients with chronic hepatitis C. Two groups, one with hyper‐gammaglobulinemia and the other without hy‐pergammaglobulinemia, were studied for the usage of TCR β chain by reverse transcription‐polymerase chain reaction (RT‐PCR) analysis. It was found that all genes of Vβ variable chain were used in the PBL of these patients. Furthermore, there was no significant difference of the TCR expression pattern between these two groups, nor a complete deletion of a particular T‐cell subset in either group. These results do not provide evidence for HCV superantigen, but indicate that the TCR usage in the patients was neither defective nor biased. © 1995 Wiley‐Liss, Inc.
Detection of hepatitis C virus‐RNA by polymerase chain reaction in dental surgeries
Authors: Piazza Marcello , Borgia Guglielmo , Picciotto Ludovico , Nappa Salvatore , Cicciarello Salvatore , Orlando Raffaele
Published: March 2007

The mean prevalence of anti‐hepatitis C virus (HCV) in Italy is 0.87%. It reaches 2% in Campania, Southern Italy. Approximately 50% of community acquired non‐A, non‐B (NANB) hepatitis cannot be associated with known parenteral exposure. A recent Italian study has shown that the only demonstrable risk factor in 9% of acute C/NANB hepatitis is dental treatment. There are no data on direct contamination by HCV of dental surgeries. Possible environmental contamination by HCV‐RNA was investigated in dental surgeries after treatment of anti‐HCV and HCV‐RNA positive patients. Thirty‐five anti‐HCV and HCV‐RNA positive patients with chronic hepatitis underwent dental treatment and were enrolled in this study. Eight had chronic persistent hepatitis (CPH), 23 chronic active hepatitis (CAH), and 4 cirrhosis. A total of 328 samples collected from instruments and surfaces were tested after dental treatment of 35 anti‐HCV positive patients. The presence of HCV‐RNA was determined by polymerase chain reaction (PCR) to evaluate contamination of instruments and surfaces in dental surgeries. Twenty (6.1%) out of 328 collected samples were positive for HCV‐RNA. The positive samples were from work benches (two), air turbine handpieces (one), holders (four), suction units (one), forceps (four), dental mirrors (two), and burs (six)

Our data indicate that there is extensive contamination by HCV of dental surgeries after treatment of anti‐HCV patients and that if sterilisation and disinfection are inadequate there is the possible risk of transmission to susceptible individuals. © 1995 Wiley‐Liss, Inc.
Evidence for lytic infection by Epstein‐Barr virus in mucosal lymphocytes instead of nasopharyngeal epithelial cells in normal individuals.
Authors: Tao Qian , Srivastava Gopesh , Chan Alexander C. L. , Chung Lap Ping , Loke Shee Loong , Ho Faith C. S.
Published: March 2007

Normal nasopharyngeal tissues from 23 individuals who died of causes unrelated to the upper respiratory system and had no evidence of Epstein‐Barr virus (EBV)‐related diseases were studied using in situ hybridisation (ISH) and immunohistochemistry for the detection of EBV RNA and expression of EBV proteins, respectively. ISH using 35S‐labelled riboprobe for EBV EBER RNA showed occasional to a few EBER + lymphocytes in the stroma of nasopharyngeal mucosa in 14/16 cases with available paraffin‐embedded tissues. In addition, very rare intraep‐ithelial EBER+ lymphocytes were also detected in 3/16 cases. However, in none of these cases was EBER detected in the epithelial cells. Similar results were obtained using a nonradioactive ISH method for EBER (Dako). In 3/23 cases, im‐munostaining using monoclonal antibodies for EBV proteins on cryostat sections showed occasional cells in the stroma expressing EBV nuclear antigen 2 (EBNA2), latent membrane protein‐1 (LMP), and switch protein encoded by BZLF1 gene (ZEBRA) in two cases and only very rare LMP+ and ZEBRA+ cells in one other case. Double immuno‐staining combining alkaline phosphatase anti‐alkaline phosphatase (APAAP) for CD markers and indirect immunofluorescence for LMP showed that the LMP+ cells were either CD19+ or less frequently CD3+, but none were CD68+. These results show that both B and T lymphocytes harbouring EBV can be found in the normal nasopharyngeal tissues. Interestingly, EBV proteins associated with lytic viral replication—diffuse early antigen (EA‐D), viral capsid antigen (VCA), or membrane antigen (MA)—were also detected in rare cells in the stroma in one case, and in another case only one MA+ cell was detected. These results provide evidence for a lytic type of infection by EBV in the normal nasopharynx involving a very small proportion of stromal lymphocytes and support the view that nasopharyngeal lymphocytes can act as a reservoir for EBV in normal individuals. © 1995 Wiley‐Liss, Inc.
Expression of the Epstein‐Barr virus DNA polymerase in Escherichia coli for use as antigen for the diagnosis of nasopharyngeal carcinoma
Authors: Lin Lung‐Shen , Ro Lee‐Hwa , Lo Mu‐Shung , Huang Wan‐Ling , Ma Jeng , Chang Tong‐Hsuan , Shu Chih‐Hung , Chow Kuan‐Chih , Liu Wu‐Tse , Chen Kuang Y. , Yang Huey‐Lang
Published: March 2007

Epstein‐Barr virus (EBV) encoded DNA polymerase (POL) was cloned and over‐expressed in Escherichia coli. Western blot analysis confirmed the presence of antibody to this POL protein in sera from nasopharyngeal carcinoma (NPC) patients. By Western blot analysis, moderate to high concentration of IgG POL‐specific antibodies were present in 43 of 48 NPC sera and only 4 of 48 healthy, seropositive controls. The POL‐specific IgG antibodies appear as early as stage I of NPC, suggesting that the recombinant POL protein can be a useful diagnostic marker for early diagnosis of the disease. It was also found that human sera containing high titer of cytomegalovirus (CMV) antibodies or herpes simplex virus type 1 (HSV‐1) antibodies did not cross‐react with the recombinant EBV POL, despite the homology shared by DNA polymerase proteins of these viruses. © 1995 Wiiey‐Liss, inc.
Controlled study of human herpesvirus 6 detection in acquired immunodeficiency syndrome‐associated non‐Hodgkin's lymphoma
Authors: Fillet Anne‐Marie , Raphael Martine , Visse Bertrand , Audouin Josée , Poirel Laurent , Agut Henri
Published: March 2007

Human herpesvirus 6 (HHV‐6) is a recently identified lymphotropic herpesvirus, which has been isolated from patients with acquired immunodeficiency syndrome (AIDS) or lymphoproliferative diseases. Two variants A and B of HHV‐6 have been described, variant B being more common in children with exanthema subitum. HHV‐6 infection was studied in cases of AIDS‐associated non‐Hodgkin's lymphoma (NHL), and in three control populations in order to evaluate the possible etiologicai role of HHV‐6 in this lymphoproliferative disease.

Tumor specimens from various organs were obtained from 27 patients with AIDS‐associated NHL and 20 human immunodeficiency virus (HlV)‐seronegative patients with NHL. Lymph node specimens were obtained from four HIV‐seropositive and nine HIV‐seronegative patients with lymph node follicular hyperplasia. A specific polymerase chain reaction (PCR) was used to detect HHV‐6 DNA. Subsequently HHV‐6 variant was identified by using variant‐specific PCR. Human cytomegalovirus (CMV) infection was detected in parallel by means of specific PCR.

HHV‐6 DNA was detected in 12 of 27 tumor tissues (44%), including 8 of 15 lymph node specimens (53%) from patients with AIDS‐associated NHL. The corresponding values in HIV‐seronegative patients with NHL were 35% (7/20) and 36% (5/14), respectively. Lymph node specimens were positive for HHV‐6 in two of four (50%) HIV‐seropositive and five of nine (55%) HIV‐seronegative patients with follicular hyperplasia. Variant A was detected in two cases of AIDS‐associated NHL, variant B in one case, and both variants in six cases. The distribution of HHV‐6 variants exhibited a similar pattern in the three control groups. CMV was only detected in 3 of 27 tumor tissues (11%) from patients with AIDS‐associated NHL.

The prevalence of HHV‐6 DNA and the distribution of its variants did not differ significantly among the four populations studied. HHV‐6 was more prevalent than CMV, a closely related herpesvirus. Most cases of HHV‐6 infection involved both HHV‐6 variants A and B. These results do not support strongly that HHV‐6 infection is closely associated with the occurrence of NHL in AIDS patients but demonstrate that mixed HHV‐6 infections are more common than previously assumed. © 1995 Wiley‐Liss, inc.
Geographic sequence variation of latent membrane protein 1 gene of Epstein‐Barr virus in Hodgkin's lymphomas
Authors: Lin Jung‐Chung , Lin Seh‐Ching , Mar Eng‐Chun , Luppi Mario , Torelli Giuseppe
Published: March 2007

To assess the role of the Epstein‐Barr virus (EBV) latent membrane protein 1 (LMP1) gene in the development of Hodgkin's lymphoma (HL), the polymorphism of this gene in EBV isolates from different geographic locations was analyzed. A 497 bp fragment spanning LMP1 gene exons 1 and 2 was amplified by polymerase chain reaction (PCR), using a primer pair bracketing a Xhol restriction site. PCR products were subjected to Xhol digestion and to DNA sequencing analysis. Twenty‐five HL biopsy specimens from the United States and five HL and four non‐Hodgkin's lymphoma (NHL) biopsy specimens from Italy were examined. Eighty percent of LMP1‐positive samples (12 of 15) from the United States maintained the Xhol restriction site and the remaining 20% partially lost the Xhol site. One of four EBV‐positive HL and one of the three EBV‐positive NHL specimens from Italy lost the restriction site. The other three EBV‐positive HL DNAs were partially cut by Xhol. Direct DNA sequencing analysis revealed that those Italian samples not digested by Xhol were due to a G to C transversion at the first base of codon 18, resulting in the change of glycine to arginine. Those DNA samples partially cut by Xhol were due to a mixture of G/C at the same location. In contrast, those partially digested American HL DNAs had a mixture of G/T at the second base of codon 17. The sequence variation found in the Italian samples differs from that of Asian EBV strains, in which G to T transversion was detected at codon 17, resulting in the substitution of arginine by leucine. Among the 72% (18 of 25) EBV‐positive American HL samples, 67% (12 of 18) were associated with type A virus, 17% (3 of 18) with type B, and 17% (3 of 18) with dual viral sequences. EBV DNA was detected in 80% (four of five) of Italian HL biopsy specimens, in which 50% (two of four) were associated with type A and 50% (two of four) with type B. Despite these sequence variations at the Xhol recognition site between EBV isolates of different geographic locations, no direct correlation with a specific genotype was observed. These results, to our knowledge, represent the first observation of a specific point mutation at codon 18 of LMP1 gene associated with a particular geographic location. It appears that the Xhol polymorphism may be a useful molecular marker for epidemio‐logic study, and the alteration in the LMP1 gene may have functional significance in the development of HL in certain geographic areas. © 1995 Wiley‐Liss, Inc.

This article is a US Government work and, as such, is in the public domain in the United States of America.
Prognostic value of adenosine deaminase compared to other markers for progression to acquired immunodeficiency syndrome among intravenous drug users
Authors: Casoli Claudio , Starcich Riccardo , Magnani Giacomo , Fiaccadori Franco , Lisa Antonella , Bertazzoni Umberto , Zei Gianna
Published: March 2007

This study was designed to determine the prognostic value of erythrocyte adenosine deaminase (ADA) as a possible indicator of progression to AIDS, and compare this with other known cellular and serological markers. At the end of a 3‐year study, a cohort of 114 human immunodeficiency virus‐1 (HIV‐1) seropositive intravenous drug users (IVDUs) from the five different Center for Disease Control (CDC) groups was examined in order to estimate the prognostic relevance with respect to the progression to acquired immunodeficiency syndrome (AIDS) of each of the following markers at baseline value: number and percentage of CD4+T cells, number of CD8+T cells, CD4+/CD8+ratio, IgA and β2microglobulin and ADA levels, and the presence of HIV antigens. Moreover, 57 IVDUs belonging to II and III CDC groups were analyzed in a follow‐up study at 6‐month intervals, in order to evaluate and compare the behavior of each marker over time. The prognostic significance of each marker was assessed by computing the survival distribution and the Cox analysis in a multivariate model providing the set of markers with greatest predictive value. The levels of ADA and the CD4+/CD8+ratio showed a linear association with disease staging, whereas β2microglobulin and CD4+/ CD8+ratio were the best predictors for AIDS progression. A highly significant increase in ADA and β2microglobulin was observed during follow‐up. The results obtained among HIV‐positive IVDUs clearly indicate that the erythrocyte ADA may be considered a reliable marker of the development of HIV infection from the intermediate stages of the disease onwards. The measurement of β2microglobulin should be regarded as a particularly useful parameter in the assessment of HIV infection in drug users. © 1995 Wiley‐Liss, Inc.
Prokaryotic expression and analysis of the antibody response to a Newcastle isolate of the core gene of hepatitis C
Authors: Milton I. D. , Carter M. J. , Toms G. L. , Watson J. P. , Bassendine M. F. , Quo K.
Published: March 2007

The full length hepatitis C virus (HCV) core gene was isolated from a Newcastle strain and expressed in E. coll. A truncated HCV core gene which lacks the hydrophobic carboxyl‐terminal sequence was also expressed. The truncated HCV core was expressed at higher levels with fewer cleavage products. Antibody reactivity to the recombinant HCV core antigen was analysed by ELISA and Western blotting in 60 HCV antibody‐positive patients with a broad spectrum of liver disease. There was no significant difference between the presence of IgG to recombinant HCV core and reactivity to the core antigen in the RIBA‐2 test. There was also no significant difference between the presence of IgG to recombinant core and diagnostic PCR as a marker for active liver inflammation. © 1995 Wiley‐Liss, inc.
Analysis of genomic polymorphism among herpes simplex virus type 2 isolates from four areas of Japan and three other countries
Authors: Sakaoka Hiroshi , Kurita Keiko , Gouro Tsutomu , Kumamoto Yoshiaki , Sawada Shigeo , Ihara Makoto , Kawana Takashi
Published: March 2007

Genomic polymorphism of 307 epidemiologi‐cally unrelated strains of herpes simplex virus type 2 (HSV‐2) from four areas of Japan and three other countries (Korea, Sweden and the U.S.A.) was analysed by using 16 variable markers selected from 97 restriction endonuclease (RE) sites with five REs. In addition to the 16 markers, 26 rare variable RE sites were found in 307 isolates. Five and four of 16 markers (RE sites) were found to differ in the frequency of isolates with the markers between isolates from Japan and Sweden and between those from Japan and the U.S.A., respectively, suggesting that they are genomically different from each other. In this manner, 307 HSV‐2 isolates from four countries could be classified into 68 different genotypes (no. of isolates/no, of genotypes = 4.5). Some isolates from one country or more than two countries at times were classified into the same genotypes, which were referred to as predominant genotypes. The most predominant genotypes for isolates from Japan, Sweden and the U.S.A. were genotypes 30, 26 (32) and 3, respectively, indicating that they are different by the country. In genotypes 1 and 6, the frequency of isolates was found to be significantly different between Japan and Sweden and between Japan and the U.S.A., respectively. Nine out of 16 markers differed in the most genomically distant isolates, each of which was obtained in Sweden and Japan. In addition, high correlation coefficients (r) in the Japanese isolates were detected in different pairs of markers from those in the Swedish isolates, suggesting that isolates from these two countries are evolutionary distant. © 1995 Wiley‐Liss, Inc.
Localization and reactivity of an immunodominant domain in the NS3 region of hepatitis C virus
Authors: Claeys Hendrik , Volckaerts Andre , Mertens Wilfried , Liang Zhong , Fiten Pierre , Opdenakker Ghislain
Published: March 2007

Analysis of the amino acid sequences of the non‐structural region 3 (NS3) of the hepatitis C virus type 1 revealed four points with a high average hydrophilicity (Ah). Two of these potential anti‐genie sites were expressed in E. coli as short fragments. The first fragment of 91 residues (NS3f3: residues 1359–1449) harbors the hexapeptide K‐K‐K‐C‐D‐E with an Ah of 2.33; the second fragment is 73 residues long (NS3f4: residues 1460–1532) and encompasses the hep‐tapeptide R‐S‐N‐R‐R‐G‐R with an Ah of 1.79. Both fragments were expressed with truncated hepatitis B core (tHBc) as a carrier protein. The fusion proteins were purified from the bacterial lysates by affinity chromatography on immobilized monoclonal antibodies against HBc, and evaluated as antigens in an enzyme immunoassay for the detection of HCV antibodies. In a specificity control panel, reactivity with NS3f3 was only found in proven HCV carriers, while reactivity with NS3f4 was weak in HCV carriers but accounted for some of the nonspecific serological reactions. In a group of 48 genotyped HCV‐in‐fected volunteer blood donors, antibodies against NS3f3 were detected in 90% (27/30) of HCV‐type 1 infections and in all HCV‐type 4 infections (5/5). However, in carriers infected with HCV‐types 2, 3, or 5, the response rate was on average only 23% (3/13). With NS3f4 as antigen, weak antibody titers were found in only about half of the carriers, independent of the infectious genotype. In a cohort of hemodialysis patients, all infected with HCV‐type 1b, NS3f3 antibodies were detected in 80% (16/20) of the carriers, while only one patient showed a weak NS3f4 reactivity. It is concluded that NS3f3 harbors an immunodominant domain of the NS3 region showing an apparently HCV‐type‐dependent serological response. © 1995 Wiley‐Liss, Inc.
Hepatitis C genotypes in hemophilic patients treated with alpha‐interferon
Authors: Devereux Helen , Telfer Paul , Lee Christine , Dusheiko Geoffrey
Published: March 2007

The effect of α‐interferon on the hepatitis C genotypes was examined in 25 anti‐HCV‐positive hemophilic patients. The rate of multiple HCV genotypes in patients who are likely to have mixed infections was also studied. Pretreatment results showed that 3/25 (12%) patients had a change in genotypes, whereas posttreatment this rose to 10/25 (40%). Seven of 10 (70%) patients who showed a change in genotype had a clinical response to α‐interferon. Six of 25 (24%) patients showed a complete clinical response to α‐interferon, and the majority of these were either type 2 or 3. This study supports previous evidence that type 1 is less likely to respond to α‐interferon, and that a‐interferon may alter the concentrations of the various circulating genotypes present in multiply‐infected patients. There are many difficulties in studying hepatitis C in hemophilic patients due to the sequence heterogeneity within each individual, and this study has shown that no ideal method exists as yet for looking at HCV genotypes in multiply‐infected individuals. © 1995 Wiley‐Liss, Inc.
Seroepidemiological study of hepatitis delta virus infection in Okinawa, Japan
Authors: Sakugawa Hiroshi , Nakasone Hiroki , Shokita Hayashi , Nakayoshi Tomofumi , Kinjo Fukunori , Saito Atsushi , Yamashiro Akihiro , Miyagi Yasuhiro
Published: March 2007

A seroepidemiological study on hepatitis delta virus (HDV) infection was conducted in the Oki‐nawan islands, the area of Japan where hepatitis B virus infection is most prevalent. The subjects of this study included 116 asymptomatic hepatitis B surface antigen (HBsAg) carriers, 48 patients with chronic hepatitis (CH), 19 with liver cirrhosis (LC), and 11 with hepatocellular carcinoma (HCC). Among the 194 serum samples examined, a total of 10 (5.2%) were anti‐HDV sero‐positive. Anti‐HDV was detected in 2 (1.7%) of the 116 asymptomatic HBsAg carriers, in 3 (6.3%) of the 48 patients with CH, and in 5 (26.3%) of the 19 with LC. However, none of the patients with HCC had detectable anti‐HDV. Eight of the 10 were born in the Miyako island group and the remaining 2 on the main island of Okinawa. Since the subjects included 34 individuals who were living and/or born in the Miyako islands, the positive rate of anti‐HDV in the islands was 23.5%. This study demonstrates the existence of an endemic area of HDV infection in Japan. © 1995Wiley‐Liss, Inc.
Precore and core mutations in HBV from individuals in india with chronic infection
Authors: Valliammai T. , Harrison Tim J. , Zuckerman Arie J. , Thyagarajan S. P.
Published: March 2007

Ten hepatitis B surface antigen seropositive carriers (5 asymptomatic and 5 with chronic liver disease) were tested for HBeAg/anti‐HBe and for HBV‐DNA using the polymerase chain reaction. Five were DNA‐positive, 2 with HBeAg and 3 with anti‐HBe. Nucleotide sequences were determined for these 5 cases. Hepatitis B virus DNA from one cirrhotic carrier with anti‐HBe had a mutation in the precore region (nucleotide position 1862) which may affect signal peptide cleavage and HBeAg synthesis. In the other 2 anti‐HBe‐ and DNA‐positive cases, a cirrhotic carrier and an asymptomatic case, there was a mutation at nucleotide position 1896 leading to a termination codon in the precore region. In all 5 patients, except for one or two missense mutations, there was no significant variation in the core region. © 1995 Wiley‐Liss, Inc.
Seroepidemiology of hepatitis E in selected Australian populations
Authors: Moaven Len , Locarnini Stephen A. , van Asten Mark , Crofts Nick
Published: March 2007

The presence of hepatitis E virus‐specific antibodies (anti‐HEV) was determined in selected Australian groups. Anti‐HEV was detected initially using a recombinant antigen‐based enzyme immunoassay (EIA). It was found that 1 of 279 (0.4%) blood donors, 14 of 182 (7.7%) Indochinese refugees, 2 of 89 (2.2%) sera submitted for amoebic serology (generally people who had visited developing countries), 1 of 13 (7.7%) patients with non‐A, non‐B hepatitis, none of 7 (0%) patients with fulminant non‐A, non‐B hepatitis, and none of 33 (0%) control sera were repeatedly reactive by the HEV EIA. The positive sera were subjected to further testing using a supplemental immunoblot. Preliminary data suggest that while potentially large numbers of people infected with HEV are entering Australia, no compulsive evidence was found in these particular groups for endemic HEV infection in Australia. This is the first seroepidemiological survey of HEV in Australia. © 1995 Wiley‐Liss, inc.
Evaluation of the polymerase chain reaction for the detection of human papillomavirus from urine
Authors: Vossler James L. , Forbes Betty A. , Adelson Mark D.
Published: March 2007

The ability to detect the presence of human pap‐illomavirus (HPV)‐DNA sequences in urine was evaluated using polymerase chain reaction (PCR). DMA was purified and extracted from urine samples, and subjected to 40 cycles of amplification using the consensus primer pair MY11 and MY09. Coamplification using the β‐globin primers, GH20 and PC04, was performed as an internal reaction control. Following assay optimization, urine samples from 22 women undergoing examination for cervical dysplasia were tested for the presence of HPV‐DNA. PCR assay results were correlated with cytologic and histo‐logic findings as well as Vira Type(tm) assay results. Overall, HPV was detected by PCR in 16 (76%) of the interpretable samples. HPV sequences were detected in 13 (87%) of the 15 specimens from women showing evidence of condylomata, dysplasia, or invasive carcinoma. HPV was detected in 3 (50%) of the women whose cytologic or his‐tologic results were either negative or showed benign atypia. Although the sample size in this study is small, our results show that HPV can be detected by PCR in a majority of individuals showing evidence of HPV infection. The method described provides a means for the clinical laboratory to detect a broad range of HPV types from using a sample obtained by noninvasive techniques. The ability to easily obtain urine would allow for increased numbers of individuals to be tested, and thus, aid in our understanding of HPV. © 1995 Wiley‐Liss, Inc.
Risk factors and prevalence of hepatitis E in German immigrants from the former soviet union
Authors: Trautwein C. , Kiral G. , Tillmann H. L. , Manns M. P. , Witteler H. , Michel G.
Published: March 2007

Worldwide there is only limited information on the epidemiology of hepatitis E virus (HEV) and its association with other hepatotropic viruses. Endemic regions have been described in some Asian countries, whereas in Europe only sporadic cases have been reported. The prevalence of HEV and a series of other viral hepatitis infections was investigated in a group of 1,025 individuals immigrating into Germany from the former Soviet Union. Serum samples were tested for anti‐HEV by a commercial enzyme immunoassay (EIA) based on recombinant proteins, and by peptide EIA and immunoblot. Risk factors and other demographic information were investigated using a questionnaire and a short interview. The overall prevalence of anti‐hepatitis E antibodies (anti‐HEV) was 2.05%. The following risk factors for HEV infection were identified : age of >65 years, resident in the south‐west part of the former Soviet Union, history of hepatitis B virus (HBV) infection, and employment in health care professions. HEV prevalence is not strikingly different from that observed in Western European countries. However, the different rates found for HEV vs. hepatitis A virus (HAV) are intriguing, since similar routes of transmission (fecal‐oral) are well documented for both viruses. Exposure to HBV is surprisingly high, and the number of hepatitis C virus (HCV)‐positive individuals was also higher than those reported from Western European areas. © 1995 Wiley‐Liss, Inc.
Human papillomavirus‐16 and ‐18 replication in esophagus squamous cancer cell lines does not require sheterologous E1 and E2 proteins
Authors: and Kazumi Togawa , Rustgi Anil K.
Published: March 2007

Human papillomaviruses (HPVs) are doublestranded DNA viruses that replicate in the nuclei of squamous epithelial cells. HPVs can be classified into high‐risk (e.g., types 16, 18, 31, and 33) or low‐risk (e.g., types 6, 11, and 30), depending on their association with benign or malignant tumors. We recently described the association of HPV‐16 and ‐18 with esophagus squamous cell cancer. HPV replication was studied in representative cell lines derived from esophagus cancers. HPV‐16 and ‐18 genomes were independently transiently transfected into HCE‐4 and HCE‐7 cell lines with and without E1 and E2 genes under heterologous promoters. Southern blot analysis demonstrated that these cell lines support viral replication. However, heterologous E1 and E2 are not required for HPV replication. These findings suggest that specific host nuclear factors in esophageal squamous epithelial cells may support HPV replication. © 1995 Wiley‐Liss, Inc.
Molecular epidemiology of hepatitis C virus infection amongst intravenous drug users in rural communities
Authors: Majid Ayaz , Holmes Roger , Desselberger Ulrich , Simmonds Peter , McKee Tom A.
Published: March 2007

The prevalence of hepatitis C virus (HCV) infection amongst a group of intravenous drug users (IVDUs) resident in West Suffolk (East Anglia, England) was investigated and compared with the prevalence of infection with hepatitis B virus (HBV) and human immunodeficiency virus (HIV). In addition, both the level of HCV persistence, as defined by detection of viral RNA, and the HCV genotypes present in this population were determined. It was found that HCV antibodies were present in 59% of those tested; by comparison 22% had antibodies to HBV and 1% antibodies to HIV. HCV RNA was found in 44% of those with HCV antibody. HCV genotype 1 was the most prevalent within this population although both genotypes 2 and 3 were also represented. © 1995 Wiley‐Liss, Inc.
Sequence comparison of echovirus type 30 isolates to other enteroviruses in the 5′noncoding region
Authors: Diedrich Sabine , Driesel Gerhard , Schreier Eckart
Published: March 2007

The genetic relationship between echovirus type 30 (E30) isolates were characterised by means of amplification of a part of the 5′noncoding region by polymerase chain reaction and direct sequencing. Later, the E30 sequences were compared with other enteroviruses. Homology between recent clinical E30 isolates from different years exceeds 98% at the nucleotide level. Comparing the E30 sequence with other enteroviruses, homology varied between 68% (coxsackievirus A24) and 93% (coxsackievirus B3). E30 appeared to have coxsackie B‐like characteristics in this genomic part. It is considered that E30 and coxsackie B viruses belong to the same enterovirus subgroup. © 1995 Wiley‐Liss, Inc.
Enzyme immunoassay using baculovirus‐expressed human calicivirus (Mexico) for the measurement of IgG responses and determining its seroprevalence in London, UK
Authors: Parker S. P. , Cubitt W. D. , Jiang X.
Published: March 2007

The use of an enzyme immunoassay (EIA) employing a baculovirus‐expressed recombinant human calicivirus (Mexico virus, MxV) for the detection of IgG‐specific antibodies is described. MxV appeared to be related antigenically to a strain of small round structured virus, SRSV/UK4/Leeds/91, which had previously been shown by solid phase immune electron microscopy (SPIEM) to be related to Snow Mountain agent (SMA). One other outbreak which occurred in San Anita, USA in 1980 and was due to consumption of contaminated water was caused by a virus antigenically related to MxV.

Volunteers and patients who developed significant IgG responses to rMxV showed anamnestic IgG responses (2 to 4‐fold) in the recombinant Norwalk virus (rNV) IgG assay. Patients and volunteers who were known to have been infected with several other strains of calicivirus/small round spherical viruses (SRSV) including NV and SRSV UK3 showed no significant antibody response to rMxV in the EIA. A seroepidemiological survey of sera from 338 children in London showed that infection with MxV occurred earlier in life than NV. Primary infections with MxV were common after the age of 6 months. Over 70% of children had evidence of infection by the age of 2 years, whereas only 12% of these children had been infected with NV. High concentrations of maternal antibody were present during the first month of life which was detected in 96% of the neonates. The results suggest that the high sensitivity of the EIA may be detecting maternal antibody throughout the first 8 months of life. © 1995 Wiley‐Liss, Inc.