bluuebunny/crossref_metadata_2025 huggingface dataset:
Oral poliovirus vaccine in the United States: Molecular characterization of sabin type 3 after replication in the gut of vaccinees
Authors: Tatem Joanne M. , Weeks‐Levy Carolyn , Mento Steven J. , Dimichele Susan J. , Georgiu Alice , Waterfield Willard F. , Sheip Bettina , Costalas Carmen , Davies Thomas , Ritchey Mary B. , Cano Francis R.
Published: March 2007

Derivatives of Sabin 3 shed from recipients of oral poliovirus vaccine in the United States (U.S.) were examined for genetic changes identified in strains excreted by vaccinees in the United Kingdom [U.K.; Evans et al., 1985; Cammack et al., 1988, Macadam et al., 1989]. Among the eight primary vaccinees studied, the duration of excretion and molecular evolution of type 3 strains varied greatly. The period of virus excretion after vaccination ranged from as few as 2 days to as many as 36 days. Nucleotide sequence analysis of viral RNAs extracted from shed virus indicated that only fifty percent of the vaccinees exclusively excreted strains in which the attenuating mutation at nucleotide 472 in the 5' noncoding region of the genome had reverted from uracil (U) to cytosine (C), the nucleotide found in neurovirulent strains. Compared to the wild‐type Leon strain, the low activity of stool isolate KW4 in a complete monkey neurovirulence test demonstrated that presence of C at 472 does not render a type 3 strain pathogenic. Conversely, anisolate was identified which efficiently replicated in monkey nervous tissue and maintained the attenuated U at 472. Oligonucleotide fingerprinting and sequence analysis of viral RNAs from stool isolates indicated that one vaccinee (KW) eventually excreted intertypic recombinant strains consistent with those reported in the U.K. studies. Unique to this study, one vaccinee (KS) excreted nonrecombinant virus possessing U at 472 for up to 21 days. The significance of the KS strain profile in relation to differences in the U.S. vaccine compared to the vaccine distributed in the U.K. and other countries is discussed.
Modulation of coronavirus‐mediated cell fusion by homeostatic control of cholesterol and fatty acid metabolism
Authors: Cervin Marguerite , Anderson Robert
Published: March 2007

Cellular susceptibility to fusion mediated by murine coronavirus (mouse hepatitis virus, MHV strain A59) was separated into lipid‐dependent and lipid‐independent mechanisms with the use of subclones and selected mutants of mouse L‐2 fibroblasts. Fusion‐resistant L‐2 cell mutants had similar cholesterol and fatty acid composition as did their fusion‐susceptible parent subclone, and were presumably deficient in a genetically mutable non‐lipid, host cell factor (e.g., fusion protein receptor). On the other hand, cellular sensitivity to virus fusion, which is known to be influenced by cell cholesterol content [Daya et al., 1988], was shown further to be modulated by homeostatic alterations in fatty acid metabolism. Cholesterol supplementation of mouse L‐2 fibroblasts or of peritoneal macrophages from MHV‐susceptible mice elevated susceptibility to viral fusion. Increased fusion susceptibility occurred in cholesterol‐supplemented L‐2 cells in the absence of any detectable alterations i n host cell fatty acid composition, thus demonstrating fusion enhancement by cholesterol alone. L‐2 cells cloned by limiting dilution in normal (not cholesterol‐supplemented) medium were found to be heterogeneous i n cholesterol content. Interestingly, high cholesterol‐containing subclones had increased levels of C‐18:0, C‐18:2, C‐20:4, and C‐22:6 and markedly reduced levels of C‐18:l fatty acids when compared to low cholesterol‐containing subclones. High cholesterol‐containing subclones did not show enhanced susceptibility to viral fusion, suggesting that homeostatic alteration of fatty acid metabolism compensated for the increased cholesterol levels and countered the normally fusion‐enhancing effect of cholesterol alone. Since these observations have potentially important consequences regarding the effects of dietary cholesterol on the severity of virus infection, we examined liver titres and pathology of normal and hypercholesterolemic mice infected with MHV. Hypercholesterolemia had no significant effect on virus replication or on liver pathology in two MHV‐ sensitive strains (Balb/c and AIJ) or in one MHV‐resistant (SJLIJ) of mice. Lipid analyses of the livers from normal and hypercholesterolemic mice showed evidence of two homeostatic mechanisms (cholesterol esterification and alteration of fatty acid composition) which likely counteracted the normally exacerbating effect of cholesterol on MHV cytopathology.
Antibody to hepatitis C virus in cryptogenic chronic liver disease
Authors: Diodati G. , Tagger A. , Bonetti P. , Ribero M. L. , Drago C. , Cavalletto D. , Grossi A. , Realdi G. , Ruol A. , Alberti A.
Published: March 2007

The prevalence of antibody to hepatitis C virus (HCV) was studied in 207 patients with chronic liver disease of unknown etiology, in relation to clinical, epidemiological and histological features. Serum antibody to C‐100 epitope of HCV was detected by ELISA in 82.6% of patients, with a significant difference compared with a group of patients with primary biliary cirrhosis (10%). The presence of anti‐HCV antibody in serum did not correlate with age, sex, histological diagnosis, and activity and duration of the disease, nor with serum anti‐HBc, used as a marker of exposure to hepatitis B virus infection. These results strongly support the view that most cases that were previously defined as cryptogenic forms of chronic liver disease are in fact related to HCV infection. There was a correlation between serum anti‐HCV antibody and history of risk for parenteral exposure or of acute hepatitis. This correlation was particularly evident for transmission by parenteral route, suggesting that HCV infection may be transmitted often by this route (36.8% among anti‐HCV antibody‐positive patients and 11.1% among anti‐HCV‐negative patients). Liver disease in patients without risk factors for parenteral transmission and with lower prevalence of anti‐HCV antibody may be caused by other as yet unidentified non‐A, non‐B (non‐C) agents or may be of nonviral origin.
Growth of human immunodeficiency virus I in cultured cells in the absence of the CD4 antigen
Authors: Morrison N. K. , McCarthy K. , Hart C. A.
Published: March 2007

The ability of a variety of epithelial, embryonal, placental, and neuronal cells to express the CD4 antigen and to be infected by human immuno deficiency virus 1 (HIV‐1) was examined. Only two (IMR‐32 and HeLa‐T4) expressed CD4 detectable by indirect immunofluorescence, and both were infectable by HIV‐1. Two others, a human laryngeal carcinoma (HEp‐2) and human colonic carcinoma (HT‐291, did not express CD4 antigen but were infectable by HIV‐I. Infection of the HEp‐2 cells was detectable four months (and 20 serial passages) later. Infection of HEp‐2 cells was not inhibited by anti CD4 monoclonal anti‐ body but was by the lectin concanavalin A. These results suggest the presence of a receptor other than CD4 can be involved in HIV‐1 infection.
Comparison of class and subclass antibody response to live and UV‐inactivated RSV administered intranasally in mice
Authors: Reuman Peter D. , Keely Scott P. , Schiff Gilbert M.
Published: March 2007

To determine the effect of viral dose and replication on the subclass antibody response to RSV, mice were immunized intranasally with different doses of live RSV (104‐106 pfu) and compared to mice given an immunizing regimen of UV‐inactivated RSV. Mice given the 106 pfu dose of live RSV and mice given the 40 p.g dose of UV‐inactivated RSV had comparable class specific antibody responses to whole RSV in serum and respiratory secretions. Serum from these two groups of mice were then compared for IgG subclass response to whole RSV. A predominance of IgG2a subclass antibody was found for both immunizing regimens, and no significant differences in subclass proportions were noted between regimens. These two regimens were then compared for serum total IgG response to RSV surface glycoproteins F and G. The serum IgG response to these glycoproteins was lower after immunization with UV‐ inactivated RSV than after live‐RSV immunization (F: P = 0.03; G: P < 0.051, even though the serum IgG response of the two groups to whole RSV was comparable. The IgG subclass re‐ sponse to surface glycoproteins was evaluated for live RSV immunization. The proportions of subclass antibody responses to glycoprotein F were comparable to the subclass response proportions to whole RSV and were not characteristic of a T‐dependent response pattern. The subclass profile for glycoprotein G was not com‐ parable to that of whole RSV but was suggestive of a T‐independent response pattern.
Direct serotyping of porcine rotaviruses using VP7‐specific monoclonal antibodies by an enzyme immunoassay
Authors: Nagesha H. S. , Holmes I. H.
Published: March 2007

Employing a serotyping EIA test using Mabs both cell culture adapted and faecal porcine rotaviruses were classified into serotypes G3, G3/5, G4, and G5. The MAbs have confirmed and extended the serotyping results obtained using polyclonal antisera. These MAbs are therefore potential reagents for serotyping of porcine ro‐taviruses. Using subgroup specific MAbs sero‐types G3, G3/5, and G5 were found to contain subgroup I antigens while G4 rotaviruses contained either subgroup II or subgroup I antigens.
Simple system for isolation of cellular and viral mutants for transformation by retrovirus
Authors: Inoue Hirokazu , Isaka Masami , Takeda Shun‐Ichi , Hakura Akira
Published: March 2007

To investigate the cellular mechanism of transformation by retroviruses, we established a system for isolation of cellular and viral mutants for transformation of a rat cell line. Mutagenized untransformed cells of this line were infected with recombinant murine retrovirus containing the src gene of Rous sarcoma virus and the selective marker gene, neo. After reaching confluence, cells transformed by the src gene tend to overgrow and die. Utilizing this property of src transformed rat cells and the selective marker gene, we could easily select untransformed cell clones containing the retrovirus genome. Expression of the src gene product in the flat clones selected was examined by in vitro assay of src kinase activity. To determine whether the mutations of these flat clones were viral or cellular, the susceptibilities of the clones to transformation were examined after superinfection with the wild‐type virus and also characterized the retroviruses recovered from these clones. With this system, two novel clones were isolated. One had a defect in viral information affecting the transformed phenotype, but still retained src kinase activity like fully transformed cells. The other showed low src kinase activity but retained wild‐type transforming virus, suggesting that a cellular gene involved in viral gene expression was mutated.
Hepatitis C virus RNA in blood donor sera detected by the polymerase chain reaction: Comparison with supplementary hepatitis C antibody assays
Authors: Widell Anders , Månsson Ann‐Sofie , Sundstrom Gunnar , Hansson Bengt Göran , Nordenfelt Erik
Published: March 2007

The low specificity of screening ELISAs for antibodies to hepatitis C virus in blood donors has called for confirmatory tests. Two types of supplementary antibody assays are available, recombinant immunoblot assays (RIBA‐1 and RIBA‐2) and an antibody consumption test referred to as a neutralization assay. Amplification of viral nucleic acid by the polymerase chain reaction (PCR) provides an antibody independent mode of detecting viral infection. We applied reverse transcription‐double PCR to detect an HCV 5'‐noncoding viral RNA sequence in serum specimens and compared PCR findings with confirmatory antibody tests. This study includes sera from 37 blood donors found positive by the Ortho anti‐HCV (C100‐3) ELISA out of 14,591 donations. Of the 37 positive sera, 8 were positive by RIBA‐1 and 1 further by RIBA‐2. Seven of the RIBA positive sera contained HCV RNA by PCR. Among the 8 indeterminate and the 21 negative donor sera by RIBA‐I, no PCR positive serum was found. The 37 anti‐HCV positive donor sera identified by Ortho ELISA were also tested by Abbott anti‐HCV (C100‐3) ELISA whereby 22 were positive. Of these 22 sera plus 1 further with ELISA OD just below cutoff, 8 were positive by the “neutralization assay,” (Abbott Laboratories, North Chicago, IL, USA) and 6 of these, including the borderline serum, were PCR positive. One of the two neutralizable but PCR negative sera was RIBA positive and the other was indeterminate. However, one ELIS. A (Abbott Laboratories) positive (OD 1.99) serum was not neutralizable but nevertheless contained HCV RNA by PCR. Thus in blood donor sera, anti‐HCV (C100‐3) reactivity confirmed by RIBA or “neutralization” correlated well with the presence of HCV RNA.
Host dependent modulation of hepatitis delta virus replication in chronic delta hepatitis
Authors: Gupta Sanjeev , Govindarajan Sugantha , Valinluck Boontar , Redeker Allan G.
Published: March 2007

To determine whether host dependent differences modulated hepatitis delta virus replication in chronic delta hepatitis, we tested HDV RNA in homosexual and intravenous drug abuser populations. Overall, the seroprevalence of HDV RNA in the two groups with matching clinical characteristics was 72% (76/106 patient visits). A trend for greater prevalence of HDV RNA was noted at initial presentation in homosexuals (82%) compared to intravenous drug abusers (6O%, P < 0.05) and this trend appeared to be maintained during two years of sequential follow‐up. The seroprevalence of co‐appearing IgM and IgG anti‐HD antibodies was similar in the two groups of patients. However, in HDV RNA positive homosexuals IgG anti‐HD antibody was more prevalent, and additionally, assumed concordance with HDV RNA of 92% although the significance of this observation is unclear. The difference in prevalence of HIV in the two groups did not reach statistical significance. Prospective studies are required to confirm differences in HDV replication in various patient groups and to define underlying mechanisms.
Levels of Pre‐S antigens and HBV DNA in sera from high and low viremic HBV carriers
Authors: Garbuglia Anna Rosa , Manzin Aldo , Budkowska Agata , Taliani Gloria , Clementi Massimo , Delfini Carlo , Carloni Guido
Published: March 2007

The biological and clinical significances of pre‐S antigens and HBV replication were investigated. Some 125 sera, 28 from HBeAg and 97 from anti‐HBe‐positive HBsAg, carriers were studied. The aim was to verify whether pre‐S antigens could be expressed in serum in complete absence of viremia. Pre‐S proteins, determined by an enzyme immunoassay, were found in sera regardless of the presence of HBV DNA, as detected by spot‐hybridization. The sera without detectable HBV DNA were investigated further by PCR using specific primers for the S and C regions of HBV. PCR analysis of samples revealed that 4 out of 5 HBeAg and 33 out of 41 (80.5%) anti‐HBe positive sera contained HBV amplified sequences of S and C regions. Pre‐S antigen values correlated well with the amounts of HBV DNA in serum detected by PCR in anti HBe‐positive subjects with high titers of pre‐S antigens (104−106). In addition, PCR highlighted the presence of HBV DNA sequences in 8 out of 17 (47.1%) pre‐S‐negative HBsAg‐positive sera.
Perinatal infection by human immunodeficiency virus type 1 (HIV–1): Relationship between proviral copy number in vivo, viral properties in vitro, and clinical outcome
Authors: De Rossi Anita , Pasti Marcella , Mammano Fabrizio , Ometto Lucia , Giaquinto Carlo , Chieco‐Bianchi Luigi
Published: March 2007

Human immunodeficiency virus type 1 (HIV‐1) isolates from 25 perinatally HIV‐1 infected children were classified according to their capacity to replicate in vitro as rapid (R), intermediate (S/R) and slow (S) variants. R‐type viruses replicated on peripheral blood mononuclear cells (PBMCs) and grew better in T‐lymphoid cells, even though 9 out of 12 isolates also maintained tropism for monocytoid cells. The S/R‐type isolates replicated efficiently after several days of culture, while the S‐type viruses displayed only a low and transient replication activity; however, both S/R‐ and S‐type isolates exerted viral transactivation activity in an indicator monocytoid cell line. Replication patterns in vitro were significantly associated in vivo with the number of HIV‐1 copies in PBMCs as determined by polymerase chain reaction: in children with R‐type isolates, the number of HIV‐1 proviral DNA molecules/105 PBMCs ranged from 62 to 571, and in children with S/R and S isolates the range was 5–43.

Seven children had severe symptomatic HIV‐1 infection, and in all an R‐type virus was identified; 18 children had no or only mild symptoms, and among these, S‐, S/R‐, and R‐type isolates were found in 5, 8, and 5 cases, respectively. Besides demonstrating HIV‐1 variability in perinatal infection, these findings suggest that R‐type virus might be a prerequisite for disease progression.
Surrogate markers are not useful for identification of HCV carriers in chronic hemodialysis patients
Authors: Willems M. , De Jong G. , Moshage H. , Verresen L. , Goubau P. , Desmyter J. , Yap S. H.
Published: March 2007

Several diagnostic hepatitis C assays have been developed for the detection of antibodies to different antigens of the virus. This virus is the major cause of non‐A, non‐B hepatitis. Seventynine patients undergoing chronic hemodialysis and/or hemofiltration were tested for the presence of anti‐HCV antibodies (anti‐C‐100‐3 antibodies and anti‐core antibodies), anti‐hepatitis B core antibodies (anti‐HBc), and aminotransferases (ALT). Seven patients were positive by one or more of the anti‐HCV enzyme linked immunoassays (EIAs), while HCV‐RNA was detectable in only four patients. These four patients had at least one, but not necessarily the same, positive anti‐HCV EIA. HCV‐RNA was not detected in patients who had no antibodies as determined by all six anti‐HCV EIAs. All patients with a marker for HCV infection had persistent normal levels of transaminases. Three patients had elevated ALT values without a marker for HCV infection and suffered from hepatitis B virus infection. Anti‐HBc was detected in 27/72 patients without any marker and in four patients with a marker of HCV infection. However, HCV‐RNA was detectable in only one of these four anti‐HBc positive patients. It is concluded that surrogate markers (anti‐HBc and serum transaminases) are not useful for identification of HCV carriers in chronic hemodialysis patients.
Occurrence and characterization of acyclovir‐resistant herpes simplex virus isolates: Report on a two—year sensitivity screening survey
Authors: Nugier F. , Colin J. N. , Aymard M. , Langlois M.
Published: March 2007

For the past 2 years, a survey network was established for the screening of acyclovir (ACV)‐resistant clinical isolates of herpes simplex virus (HSV). Among 889 strains tested for in vitro ACV sensitivity, 14 HSV‐1 and 6 HSV‐2 were resistant to ACV concentrations exceeding 3 μg/ml. These resistant isolates were most often obtained after prolonged ACV treatment of severely immuno‐compromised patients. For five patients, the emergence of ACV‐resistant virus correlated with treatment failure. In particular, a decrease in the in vitro sensitivity to ACV was observed for eight successive HSV‐1 isolates from one immunodeficient patient undergoing therapy. All ACV‐resistant isolates were studied for their sensitivity to different antiherpetic compounds and showed various cross‐sensitive and ‐resistant patterns. The examination of viral populations by plaque autoradiography procedures frequently revealed their heterogeneity in terms of thymidine kinase (TK) phenotype and allowed the detection of various proportions of TK‐positive (TK+), TK‐deficient (TKD), or TK‐altered (TKA) viruses. Our data underline the importance of monitoring the emergence of drug‐resistant virus during the course of antiviral therapy, and the need for the detection and characterization of TK mutants in clinical specimens. The routine examination of drug sensitivity of HSV isolates provides useful information to clinicians for the management of ACV treatment in the hope of preventing ACV‐resistant mutants from becoming predominant in mixed viral populations.
Significance of serum IgM anti‐HBc in chronic hepatitis B virus infection
Authors: Smith Hather. M. , Lau Johnson Y. N. , Davies Susan E. , Daniels Helena M. , Alexander Graeme J. M. , Williams Roger
Published: March 2007

Because of widely differing reports on the significance of IgM anti‐HBc in chronic hepatitis B virus (HBV) infection, paired sera and liver biopsies from 49 patients with chronic HBV infection were analysed for serum IgM anti‐HBc, HBsAg titre, HBeAgianti‐HBe, HBV DNA, serum aspartate transaminase, intrahepatic HBcAg expression, and liver histology. High levels of IgM anti‐HBc, i n the diagnostic range of acute hepatitis B (>1.2), were detected in seven patients (14.3%) and a total of 34 patients (69.6%) had an index of more than 0.2. No correlation was found between IgM anti‐HBc and the serum markers of active viral replication or HBsAg titre but it correlated significantly with intrahepatic expression of cytoplasmic HBcAg (r2 = 0.165, P =0.002). IgM anti‐HBc also correlated with active liver histology (P = 0.015) but there was a considerable overlap of the IgM anti‐HBc index values between the various disease groups, indicating a poor specificity. Serial assessment of IgM anti‐HBc in eight patients treated with interferon‐α (four responders) showed an increase in IgM anti‐HBc in three out of four patients corresponding to the e‐seroconversion period followed by a drop in IgM anti‐HBc levels. However, an increase in IgM anti‐HBc was also seen in one non‐responder, indicating that this feature is not unique to interferon‐α responders. These data indicate that serum IgM anti‐HBc cannot be used alone as a certain diagnostic measure of HBV replication nor in the prediction of liver histology.
Comparison of hepatitis B virus subtyping of d/y determinants by radioimmunoprecipitation assay and the polymerase chain reaction
Authors: Nicholson W. J. , Black S. H. , Simmonds P. , Chung C‐W. , Aw D. , Peutherer J. F.
Published: March 2007

Using a double polymerase chain reaction a method was devised for detecting and subtyping hepatitis B virus DNA in serum samples. Primers from the S‐gene were selected from the sequence analyses of five HBV HBsAg subtypes, to amplify HBV DNA and subtype for y specific DNA. Thirty‐eight samples were subtyped for dand y determinants by radioimmunoprecipitation assay (RIPA) and the polymerase chain reaction (PCR). Subtyping by PCR and RIPA was in agreement in 100°/o of subtype y samples and 83.3% of subtype d, giving an overall correlation of 92.1%. As a third comparison, 12 amplified samples were digested by the restriction enzyme Sau 3A, which differentiates between subtypes yand d. The digest results agreed with PCR in 83.3% of the samples. In addition, we compared our standard phenol/chloroform extraction against a rapid one step method. The phenol/chloroform stage was found to be essential for the removal of nucleases and polymerase inhibitors present in sera.
The diagnostic significance of the polymerase chain reaction and isoelectric focusing in herpes simplex virus encephalitis
Authors: Pohl‐Koppe Annette , Dahm Cornelia , Elgas Michael , Kühn Joachim E. , Braun Rüdiger W. , Meulen Volker te.
Published: March 2007

In this study, serum and CSF samples of 55 neurological patients have been examined to confirm the diagnosis of herpes simplex virus encephalitis (HSVE). Different methods were applied, including serological titer evaluations, determination of intrathecally‐produced HSV‐specific antibodies by isoelectric focusing with affinity immunoblotting (IEF), as well as HSV‐specific ELISA and HSV‐specific polymerase chain reaction (PCR). The results of IEF and PCR have been compared and contrasted to develop general directions for virological diagnosis of HSVE. Of 14 patients suffering from clinically diagnosed HSVE, HSVE was confirmed in 12 cases by the demonstration of PCR or IEF positivity. A HSV‐specific CNS infection could be excluded in 2 of these 14 patients. In 17 patients suffering from non‐HSVE, PCR and IEF results were negative. Twenty‐four patients, suffering from other neurological diseases, serving as a control group, were PCR‐ and HSV‐IEF‐negative. The study indicated that there are two possibilities for unequivocal demonstration of HSV‐specific CNS involvement: first, performance of PCR especially in the acute phase of disease and in sus‐picious relapses, and second, performance of HSV‐specific IEF for determination of intrathecally synthesized HSV‐specific antibodies. It is suggested that these two methods should be introduced in routine diagnosis of viral encephalitis.
Herpes simplex virus infection of cultured human term trophoblast
Authors: Nørskov‐Lauritsen N. , Aboagye‐Mathisen G. , Juhl C. B. , Petersen P. M. , Zachar V. , Ebbesen P.
Published: March 2007

Mononuclear trophoblast cells were isolated from term placentas of uncomplicated pregnancies, purified to homogeneity by negative immunomagnetic separation using monoclonal antibodies to the major histocompatibility complex, and challenged with herpes simplex virus (HSV). The cultures were highly susceptible to virusinduced cytopathic effect as evidenced by cytopathic alteration and inhibition of human chorionic gonadotropin (hCG) secretion. Both HSV I and II underwent a full replicative cycle in the trophoblast, although the production of progeny virus was 10‐100 times less than that obtained with placental fibroblasts or choriocarcinoma cells. The permissiveness was independent of in vitro syncytial differentiation of the trophoblast. The results suggest that the trophoblast layer may be involved in intrauterine HSV infection.
Early diagnosis of enteroviral meningitis by detection of specific IgM antibodies with a solid‐phase reverse immunosorbent test (SPRIST) and μ‐capture EIA
Authors: Glimåker Martin , Samuelson Agneta , Magnius Lars , Ehrnst Anneka , Olcén Per , Forsgren Marianne
Published: March 2007

A solid‐phase reverse immunosorbent test (SPRIST) and a μ‐capture enzyme immunosorbent assay (EIA) for detection of enterovirus‐specific IgM antibodies were evaluated for enterovirus diagnosis of aseptic meningitis in 160 consecutive patients from whom enterovirus (11 different serotypes) were isolated in 64. In patients with an enterovirus isolate and/or four‐fold titre rise in the complement fixation test (CFT) for enterovirus, specific enterovirus IgM antibodies were detected on the day of admission to hospital in 48% by SPRIST and in 50% by EIA and 4‐6 days after onset of symptoms in 71% by SPRIST and 79% by EIA. A significant increase in titre was observed between serum sampled on the day of admission and 2 days later in 38% by SPRIST and in 41% by EIA. These results indicate that the IgM antibody response appears early in the course of aseptic meningitis. Since both SPRIST and EIA provide rapid results the tests may be of differential diagnostic value and the IgM antibody kinetics may be utilized for diagnosis during the acute phase of aseptic meningitis. With optimized serum sampling the positive outcome was 76% in SPRIST and 82% in EIA among patients with positive virus isolation and/or CFT for enterovirus.

In 67 patients virus isolation and CFT for en‐terovirus yielded negative results as well as all non‐enteroviral diagnostic tests. Thirty‐eight of these patients were positive by SPRIST and/or EIA and in half of these 38 a significant titre rise and/or fall in SPRIST and/or EIA was recorded. The majority of these IgM‐positive patients became ill in the late summer or autumn, i.e., the “enterovirus season.” These findings indicate that SPRIST and EIA may give additive information about etiology in patients with aseptic meningitis. However, enterovirus IgM responses were seen in 10 cases by SPRIST and in 6 by EIA out of the 25 patients with non‐enterovirus diagnoses. This indicates that the tests may be not only cross‐reacting within the enterovirus group but also, to a smaller extent, be cross‐reacting with sera from patients with non‐enterovirus infections. Dual infections or persistent enterovirus IgM activity after a recent enterovirus infection may be alternative explanations. Thus, additional criteria in support of present enterovirus infection should be sought. In conclusion the detection of enterovirus‐specific IgM has improved the early diagnosis of aseptic meningitis, but rapid enterovirus diagnosis still remains a challenge.
A high frequency of human papillomavirus DNA sequences in cervical carcinomas of indian women as revealed by southern blot hybridization and polymerase chain reaction
Authors: Das B. C. , Sharma J. K. , Gopalkrishna V. , Das D. K. , Singh V. , Gissmann L. , Hausen H. Zur , Luthra Usha K.
Published: March 2007

Ninety‐six colposcopical ly directed biopsies from squamous epithelial carcinoma of the uterine cervix and 22 age‐matched normal control biopsy specimens were examined by both Southern blot hybridization and polymerase chain reaction (PCR) for the presence of different human papillomavirus (HPV) DNA types. Cancer of the uterine cervix, which is the most common malignant disease in Indian women, showed a high frequency (98%) of HPV as compared to those reported from other parts of the world. HPV type 16 was found to be the dominant (64%) type while the frequency of HPV type 18 was verylow (3%). On individual typing of HPV, no biopsy was found to contain any other known HPV types under stringent conditions of hybridization except a single case of HPV type 11. Only one case of double infection with HPV types 16 and 18 was recorded. Under low stringency conditions of hybridization with a mixed probe of HPV types 16 and 18, 29 additional biopsies were found to be positive. Southern blot hybridization alone detected HPV DNA in 92% of the cases but none in the controls. By PCR, six (6.25%) more cases and four (18.18%) healthy women were found to be positive for HPVs. Analysis of the physical state of HPV 16 indicated integration in about 70% of carcinoma cases while 30% of them were in episomal form. The findings suggest that infection with HPV is an important etiologic factor for the development of cervical cancer, that a number of such tumours may arise without HPV infection, and that integration of the viral DNA into host genome is not always essential for malignant progression. Furthermore, the use of PCR provides an effective complementation of Southern blot hybridization for meaningful epidemiological studies of HPV infection.
A pilot model of vaccination against hepatitis B virus suitable for mass vaccination campaigns in hyperendemic areas
Authors: da Villa G. , Piazza M. , Lorio R. , Picciotto L. , Peluso P. , Deluca G. , Basile B.
Published: March 2007

A hepatitis B vaccination campaign was carried out in a town of 60,000 inhabitants, Afragola, Campania, Italy, a hyperendemic area for hepatitis B where HBsAg prevalence was 13.4% and anti‐HBc prevalence was 64.7%. This experimental pilot project aimed to reduce the incidence of both acute and asymptomatic viral hepatitis B and of related chronic liver complications. From 1983‐1989, 8,400 subjects were vaccinated: 6,900 children up to 10 years of age and 1,500 subjects from 11‐60 years of age. High seroconversion rates were observed: 99.0% in all children under one year of age, 96.0% in the older children, and 86.7% in adults. The rate of infection in Afragola has diminished from 63/100,000 in 1983 to 10/100,000 in 1989. Carriers of HbsAg decreased in the general population (7.3% com‐ pared to 13.4%). especially in children up to 10 years of age (1.0 compared to 9.0%). In babies who received hepatitis B vaccine at the same time as compulsory vaccinations compliance was 98% while it was 80% in babies who were vaccinated separately.

In June 1991 the Italian Parliament promulgated a decree which imposes hepatitis B vaccination for all newborn babies at 3, 5, and 1 1months of age, at the same times as the mandatory childhood accinations (diphtheria, tetanus, and polio) according t o a new protocol (Piazza scheme) which has been in use since January 1987 in our pilot vaccination campaign in Afragola.
Molecular epidemiology of adenoviruses associated with acute lower respiratory disease of children in Buenos Aires, Argentina (1984–1988)
Authors: Kajon Adriana E. , Wadell Göran
Published: March 2007

DNA restriction analysis was carried out on a sample of 73 adenovirus strains isolated in Buenos Aires from nasopharyngeal aspirates of children with lower acute respiratory infection between 1984 and 1988.

Thirty‐five isolates (47.9%) were classified as members of subgenus B. Of these, three were identified as a new genome type of Ad3p denominated Ad3p3; five strains corresponded to genome type 7b and two to genome type 7c. The other 25 isolates were identified as the recently recognized genome type 7h. All 6 fatalities re‐ corded within this group of 73 children were associated with infection by Adenovirus genome type 7h.

Thirty‐seven isolates (50.7%) were classified within subgenus C that corresponded to 9 different genome types denominated 1p (n = 5); I#(n = 2); 2p (n = 4); 2b (n = 6); 2# (n = 5); 5# (n = 4); 5* (n = 7) and 5+ (n = 2). All genome types of subgenus C were compared with the data reported by Adrian et al. (Archives of Virology112:235‐238,1990). The Ad1p and Ad1# genome types could be allocated to AVI genome types D1 and D10, respectively. Ad2b genome type could be allocated to AV2 genome type D25. No counterparts were found for the remaining 6 genomic variants.

Only one isolate was identified as Ad4a of subgenus E.

The comparison of the results of the present study with those of the molecular characterization of Chilean strains isolated between 1984 and 1987 suggests that the adenovirus strains associated with respiratory disease of children may be common in both countries.
Outcome of herpes simplex virus type 2 infection in guinea pigs
Authors: Fowler Sandra L. , Harrison Christopher J. , Myers Martin G. , Stanberry Lawrence R.
Published: March 2007

Factors that influence the outcome of genital herpes simplex virus (HSV) infection were explored in a guinea pig model. The viral inoculum required to establish infection in 50% of animals (ID50) was similar for inbred (strain 2) and out‐bred (Hartley) guinea pigs. However, the viral inoculum required to produce clinical disease in 50% of the animals (CD50) was 10 times greater for strain 2 compared to Hartley animals. HSV infection of both inbred and outbred animals was more likely to result in death of weanling than adult animals. The duration and severity of genital disease and the magnitude of vaginal viral replication were similar for strain 2 and Hartley animals in both young and adult animals. The lethal dose for 50% of animals (LD50) was 100‐ fold greater than the CD50 for Hartley animals, but the LD50 and the CD50 were equal in strain 2 guinea pigs. Viral cultures of homogenized neural tissues from infected animals revealed that HSV ascended to the level of the temporal cortex in strain 2 guinea pigs while virus was never recovered above the lumbar spinal cord in Hartley animals. Endogenous peripheral blood mononuclear cell‐mediated cytolytic activity against HSV‐infected targets was greater prior to HSV inoculation in survivors compared to animals that died.

A fatal outcome of genital HSV‐2 may relate to the failure to limit CNS viral replication. Death is more common among guinea pigs that have low endogenous HSV‐directed natural killer activity, such as occurs among strain 2 and young animals whether inbred or outbred.
Effect of cyclosporin‐A in woodchucks with chronic hepatitis delta virus infection
Authors: Karayiannis P. , Goldin R. , Luther S. , Carman W. F. , Monjardino J. , Thomas H. C.
Published: March 2007

Four woodchucks chronically infected with hepatitis delta virus (HDV) were treated with cyclosporin‐A (CyA) for 11 weeks. All animals had detectable HDAg in the liver and two of them were also positive for HDAg and HDV‐RNA in serum. Reappearance of HDV in serum was noted in one of the two non‐viraemic animals and increased viraernia in the two viraemic. HDV‐RNA levels became elevated within a week of starting treatment and an inverse relationship between HDV‐RNA and WHV‐DNA became apparent during the treatment period. With discontinuation of treatment, HDV‐RNA levels either returned to pretreatment levels or became negative. The remaining animal showed no return of viraemia during CyA treatment; HDV‐RNA remained negative and WHV‐DNA levels did not change. Liver biopsies revealed a slight increase in lobular activity during CyA treatment in the animals showing increased viraemia. These data are consistent with the hypothesis that the host immune response exerts a negative control on the level of HDV viraemia and that HDV influences HBV replication independently of the host immune response. In an animal that may have been clearing HDV, immunosuppression did not result in recurrence of viraemia.
Inactivated Lassa virus elicits a non protective immune response in rhesus monkeys
Authors: McCormick J. B. , Mitchell S. W. , Kiley M. P. , Ruo S. , Fisher‐Hoch S. P.
Published: March 2007

We attempted to protect three rhesus monkeys from Lassa fever by vaccination with a preparation of purified whole Lassa virus which had been inactivated by gamma irradiation. The vaccinated monkeys developed antibodies against the three major viral proteins of Lassa virus demonstrated by radioimmunoprecipitation. When the three vaccinated monkeys and two unvaccinated control monkeys were challenged all five became severely ill and died. Prior to death a secondary, high‐titer antibody response to Lassa virus was observed in the three vaccinated monkeys, whereas the two unvaccinated monkeys developed a primary, low‐titer antibody response. Though titers of Lassa virus in serum reached peak levels earlier following challenge in the non vaccinated, at the time of death serum and organ virus liters did not differ significantly. Changes in platelet aggregation, leukocyte counts, and liver enzymes, abnormalities of which have been associated with severity of Lassa fever, were found to be comparable in the two groups. The humoral antibody response measured in these animals following vaccination, although of the same magnitude as found in humans recovered from Lassa fever, was insufficient to protect the animals from this fatal disease. Evidence is now accumulating that the cell‐mediated immune response must be activated in order to protect against challenge with arenaviruses. © 1992 Wiley‐Liss, Inc.
Prevalence of anti‐HCV and HCV viremia in hemodialysis patients in Taiwan
Authors: Sheu Jin‐Chuan , Lee Suh‐Huoy , Wang Jin‐Town , Shih Linn‐Na , Wang Teh‐Hong , Chen Ding‐Shinn
Published: March 2007

Hepatitis C viral infection in 125 hemodialysis patients from Taiwan was studied using a sec‐ ond‐generation anti‐HCV immunoassay (EIA 11) (Abbott HCV 2.0 EIA) and the polymerase chain reaction (PCR) to detect the HCV RNA in the se‐ rum. A total of 59 patients (47.2%) were positive by EIA II. In comparison, the conventional CIOO‐3 anti‐HCV assay was positive in 40 (32.0%). HCV RNA was found in 47 patients (37.6%). Patients with elevated serum transaminase level had a higher positive rate of anti‐HCV and HCV RNA. The dialysis time was longer for those patients positive for anti‐HCV than for those who were negative. A total of 57 of the 59 EIA Il‐positive cases had a history of blood transfusion. The HBsAg status did not influence the anti‐HCV positivity. Among the 59 EIA Il‐positive patients, 66.1% were also positive for HCV RNA, and of the 47 HCV RNA‐positive cases 83.0% were positive for EIA 11. It is concluded that the high prevalence of specific HCV infection and HCV viremia was present in these patients. Prevention of cross‐ contamination during dialysis and blood screen‐ ing before transfusion are important for the control of HCV infection in these patients. © 1992 Wiley‐Liss, Inc.
Effects of 2′‐fluorinated arabinosyl‐pyrimidine nucleosides on duck hepatitis B virus DNA level in serum and liver of chronically infected ducks
Authors: Fourel I. , Li J. , Hantz O. , Jacquet C. , Fox J. J. , Trepo C.
Published: March 2007

The 2′‐fluorinated arabinosyl‐pyrimidine nucleo‐ sides, 1‐(2′‐deoxy‐2′‐fluoro‐P‐D‐arabinofura‐ nosyl)‐5‐iodocytosine (FIAC) and 1‐(2′‐deoxy‐ 2′‐fluoro‐p ‐D‐arabinofuranosyl) ‐5‐methyluracil (FMAU), are new antiviral compounds with in vitro inhibitory activity against the DNA polymerase of hepadnaviruses. Those compounds also induced permanent inhibition of viral replication in woodchucks chronically infected by woodchuck hepatitis virus. The effects of these antiviral compounds were assessed in ducks chronically infected by duck hepatitis B virus (DHBV). Following intraperitoneal administration for 5 days, FMAU (2 mgikgiday) and FIAC (10 mgikgiday) induced a transient decrease in DHBV replication, as shown by the decrease in both the serum and liver DHBV DNA level. After stopping therapy, DHBV replication rebounded immediately to the pretreatment level. The supercoiled form of liver viral DNA was found to be less affected by the therapy. By contrast, no obvious antiviral effect was observed with vidarabine monophosphate (ara‐AMP) (80 mglkgiday) therapy. No sign of toxicity was observed during the course of the treatment. These preliminary results confirmed in the DHBV model the higher efficacy of FIAC and FMAU as compared to ara‐ AMP. Pharmacokinetic studies are needed to ex‐ plain the differences observed in viral replication in these 2 models of HBV infection. © 1992 Wiley‐Liss, Inc.
Detection of cytomegalovirus‐infected cells by flow cytometry and fluorescence in suspension hybridisation (FLASH) using DNA probes labeled with biotin by polymerase chain reaction
Authors: Link Hartmut , Battmer Karin , Kleine Hans Dieter
Published: March 2007

A biotin‐labeled DNA probe specific for the im‐ mediate early gene of the cytomegalovirus (CMV) strain AD 169 was generated with the polymerase chain reaction. Nucleic acid hybridisation was carried out with CMV‐infected T‐lymphoblastoid cells (MOLT4) after 4 hr to 6 days of culture. Biotin molecules were made visible with streptavidin coupled with fluorescein. The fluorescence signal of the hybridised probe was measured by flow cytometry in the cell suspension. The number of CMV‐positive cells was 7% at 4 hr, 8% after 28 hr, 18% after 2 days, 26% after 3 days, 91% after 4 days, 97% after 5 days, and 98% after 6 days. The first detection of CMV antigen (pp65) was possible with immunoenzymatic labeling by day 4, whereas CMV‐DNA was detected by PCR after 4 hr. CMV‐specific RNA could be detected in a similar way. The analysis of mononuclear peripheral blood leukocytes in a patient with active CMV infection showed 14.7% CMV DNA‐positive cells at day 1 and 7% at day 8, as compared to 0.9% and 0.0% cells which were positive for CMV antigen (pp65) by immunoenzymatic labeling at day 1 and day 8, respectively. We conclude that flow cytometry and fluorescence in suspension hybridisation (FLASH) offers a new tool for analysing exactly and quantifying large numbers of cells for specific DNA or RNA and may be useful for other laboratory and clinical applications. © 1992 Wiley‐Liss, Inc.
Improved serodiagnosis of chronic hepatitis C in Japan by a second‐generation enzyme‐linked immunosorbent assay
Authors: Yuki Nobukazu , Hayashi Norio , Hagiwara Hideki , Takehara Tetsuo , Oshita Masahide , Kasahara Akinori , Fusamoto Hideyuki , Kamada Takenobu
Published: March 2007

To clarify the role of hepatitis C virus (HCV) infection in chronic liver disease, sera from Japanese patients which were negative by the original anti‐HCV assay (Ortho) were subjected to a second‐generation anti‐HCV assay based on a combination of structural (C22) and nonstructural (C200) recombinant HCV proteins. Of 29 patients with chronic non‐A, non‐B hepatitis, 20 (69%) were anti‐HCV‐positive by the second‐generation enzyme‐linked immunosorbent assay (ELISA) and also positive by the reverse transcription‐polymerase chain reaction (RT‐PCR) which detects the HCV genome. Of 41 chronic hepatitis B virus carriers, 3 (7%) were positive by the second‐generation ELISA; 1 of 3 was positive by RT‐PCR. The HCV genome was detected in ail cases positive for anti‐HCV with high titers. Of 59 healthy subjects negative by the second‐generation ELISA, none were positive by RT‐PCR. These findings indicate that HCV is a major causative agent of chronic non‐A, non‐B hepatitis in Japan and that second‐generation ELISA is specific and a more sensitive diagnostic assay for chronic hepatitis C. © 1992 Wiley‐Liss, Inc.
Antigenic relationships among human herpesvirus‐6 isolates
Authors: Chandran Bala , Tirawatnapong Suranan , Pfeiffer Brian , Ablashi Dharam V.
Published: March 2007

Human herpesvirus 6 (HHV‐6) prototype isolate GS is a newly identified lymphotropic herpesvirus and several subsequent herpes isolates were recognized as HHV‐6 by their hybridization to a HHV‐6(GS) DMA probe pZVH14. DNA restriction analysis and in vitro tropism studies show that HHV‐6 isolates can be divided into two groups, designated group A and group B. Antigenic relationships among 15 HHV‐6 isolates belonging to these two groups were examined using rabbit antibodies against HHV‐6(GS) infected cells, 11 monoclonal antibodies against three glycoproteins and four non‐glycoproteins of HHV‐6(GS), and sera from 136 healthy adults. More than 20 polypeptides from all these isolates were immu‐noprecipitated by rabbit polyclonal antibodies against HHV‐6(GS) infected cells. Reactivities of monoclonal antibodies segregated these isolates into the same two groups. Group A contains HHV‐6(GS), HHV‐6(U1102) from a Ugandan acquired immunodeficiency syndrome (AIDS) patient, and nine other HHV‐6 isolates from various disorders. HHV‐6(Z‐29) from a Zairian AIDS patient, HHV‐6(SF) isolated from the saliva of a human immunodeficiency virus (HlV)‐infected individual, HHV‐6(OK) from a child with exan‐them subitum, and HHV‐6(DC) from a leukopenia patient are in group B. Eighty‐one percent of the sera showed similar antibody titer in immunofluorescence assay with group A HHV‐6(GS) and group B HHV‐6(Z‐29) infected cells and 19% of the sera showed two‐ to four‐fold antibody titer differences. The mobilities of many of the polypeptides immunoprecipitated from group A HHV‐6(GS) and group B HHV‐6(Z‐29) infected cells were different and sera showed differences in the quantities and nature of polypeptides immunoprecipitated. Together, our data show that HHV‐6 related isolates segregate into two antigenically closely related yet distinct groups, exhibiting group common and group specific anti‐genie epitopes. The complex patterns of reactivities of human sera suggest that individuals may be infected with viruses from one group. © 1992 Wiley‐Liss, Inc.