bluuebunny/crossref_metadata_2025 huggingface dataset:
Prospective study on maternal, intrauterine, and perinatal infections with cytomegalovirus in Japan during 1976–1990
Authors: Hirota Kiyokata , Muraguchi Kiyo , Watabe Noritsune , Okumura Masayuki , Kozu Masaomi , Takahashi Kenji , Machida Yukio , Funayama Yuko , Oshima Takeko , Numazaki Yoshio
Published: March 2007

Since 1976, sera obtained serially from 10,218 pregnant women during the first, second, and third trimesters of gestation and cord sera were tested for CMV complement‐fixing (CF) and im‐munofluorescent (IF) antibodies. CMV IgG‐IF antibody was positive in 9,735/10,218 (95%) in the first trimester, and a significant rise of CF antibodies during pregnancy was found in 70/9,206 (0.76%) of the seropositive group and in 5/438 (1.14%) of the seronegative group. IgM antibody was found in 6/9,206 (0.06%) of seropositive women during the first trimester and in 7/70 (10.0%) of seropositive mothers with CF antibody rise and in 4/5 of seroconverted mothers of the seronegative group, suggesting that the incidence of primary infection with CMV during pregnancy was approximately 1% of susceptible women. All the mothers with immune response had infants with neither viruria nor IgM antibody in the cord blood, whereas seropositive mothers without an immune response had infants with viruria (7/1,826; 0.4%) or with IgM antibody in the cord blood (6/9,136; 0.06%). None of these 13 babies, shedding CMV or with IgM IF antibody, had physical or mental retardation. CMV IgG‐IF antibody was present in almost 80% of infants between 7 and 12 months of age in 1988, suggesting that perinatal or postnatal CMV infection may occur in infants born to seropositive mothers in 70‐80% of pregnancies. © 1992 Wiley‐Liss, Inc.
Sexual transmission of human T‐lymphotropic virus type I in Peruvian prostitutes
Authors: Wignall F. Stephen , Hyams Kenneth C. , Phillips Irving A. , Escamilla Joel , Tejada Abelardo , Li Olga , Lopez Francisco , Chauca Gloria , Sanchez Sara , Roberts Chester R.
Published: March 2007

The epidemiology of HTLV‐I infection in female prostitutes was studied in a survey of 395 prostitutes from Callao, Peru (the port city of Lima), 72 prostitutes from Iquitos, Peru (another port city on the Amazon River), and 510 prenatal clinic patients from Lima. Prostitutes reported a mean of 8.8years (range, 1–39 years) of active prostitution and a mean of 205 sexual contacts during the month prior to the study. The percentage of prostitutes with HTLV‐I antibody (21.8%) was significantly higher than patients attending a prenatal clinic (3.1%; P < .0001). The prevalence of HTLV‐I antibody increased steadily with age in prostitutes, but no age trend was noted i n prenatal patients. By multiple logistic regression analysis, an independent association was found between HTLV‐I seropositivity and a history of prostitution i n Callao, age, and positive syphilis serology when all 977 study subjects were evaluated. When prostitutes alone were analyzed, the number of years of exposure as a practicing prostitute was associated with HTLV‐I seropositivity after controlling for age. These data indicate a greatly increased risk of HTLV‐I infection in prostitutes in Callao, Peru, and suggest an association between sexual activity and HTLV‐I transmission. © 1992 Wiley‐Liss, Inc.
New developments in the molecular epidemiology of adenovirus 8 keratoeonjunctivitis
Authors: de Jong Jan C. , Démazure Marc , Legrand‐Quillien Marie‐Christine , Le Lay Geneviève , Colin Joseph , Wermenbol Anton G. , Verweij‐Uÿterwaal Marja W. , van der Avoort Harrie G.A.M. , Chastel Claude
Published: March 2007

Four consecutive epidemics of keratoconjunctivitis caused by adenovirus 8 (Ad81 occurred over a 5‐year period in Brest, France. A selection of 30 strains isolated during this period was studied by DNA restriction enzyme analysis using nine restriction enzymes. Bgll and Sac1 were the most discriminative enzymes and allowed the recognition of four DNA variants, all different from the prototype strain Trim. Within each of the epidemics, the strains tested could not be distinguished in this analysis. Between strains from different epidemics differences in DNA structure could be detected however. Thus, the Ad8 epidemics of 1983/1984, 1984, 1987, and 1988 appear to have been due to DNA variants Ad8/D7, D8, D9, and D10, respectively. These results demonstrate that the DNA of Ad8 seems to display a considerable variability, comparable to that observed with Ad7 and Ad21. As has been described for Ad7, Ad21 and Ad41, successive DNA variants of Ad8 prevail during one or more years, and are then replaced by other, newly emerging variants sometimes associated with epidemics. © 1992 Wiley‐Liss, Inc.
Characterization of Dobrava virus: A hantavirus from Slovenia, Yugoslavia
Authors: Avsic‐Zupanc Tatjana , Xiao Shu‐Yuan , Stojanovic Radivoj , Gligic Ana , van der Groen G. , Leduc James W.
Published: March 2007

Small mammals were collected in natural foci of hemorrhagic fever with renal syndrome (HFRS) in Slovenia, Yugoslavia, and a hantavirus was isolated from the lungs of an Apodemus flavicol lis captured in Dobrava village. This new isolate, Dobrava virus, was compared with representative strains of the Hantavirus genus by serological and polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) methods. It was found by cross immunofluorescent and enzyme‐linked immunosorbent assays that antigenic properties of Dobrava virus were different from those of other hantaviruses. The RNA of this virus was successfully amplified with hantavirus genus reactive primer sets by reverse transcriptase polymerase chain reaction (RT‐PCR); however, PCR‐RFLP analysis of the amplified product was shown to be unique among those of the known hantaviruses, further indicating that Dobrava virus represents a new hantavirus serotype. Published 1992 Wiley‐Liss, Inc.
Rapid detection of respiratory viruses using mixtures of monoclonal antibodies on shell vial cultures
Authors: Schirm Jurjen , Luijt Dirk S. , Pastoor Geke W. , Mandema Johan M. , Schröder F. Peter
Published: March 2007

Eleven hundred and thirty‐three clinical specimens submitted to the laboratory for diagnosis of respiratory virus infections were tested by direct immunofluorescence (DIF) for respiratory syncytial virus (RSV), by shell vial culture, and by conventional cell culture. The shell vial cultures were stained with 8 different monoclonal antibodies both 1 day and 3–7 days after inoculation. In order to limit the cost and the workload, mixtures of monoclonal antibodies were used. Coverslips with HEp−2, cells were incubated with a mixture of FITC‐labeled monoclonal antibody to RSV and nonlabeled monoclonal antibody to adenovirus. When no RSV positive IF staining was observed after the first incubation step, the same coverslip was incubated once more with FITC‐ labeled anti‐mouse antibody. A positive reaction at this stage indicated the presence of adenovirus. Similarly, cultures of tertiary monkey kidney cells were investigated with a mixture of two FITC‐labeled monoclonals to the influenza viruses A and B and three nonlabeled monoclonals to the parainfluenza viruses 1, 2 and 3. If influenza virus or parainfluenza virus was detected, the exact type was determined by staining different parts of a duplicate coverslip. Shell vial cultures for cytomegalovirus (CMV) were always performed separately on human embryonic lung fibroblasts.

Using this approach, we detected RSV (n = 248), CMV (n = 42), parainfluenza virus (n = 31), influenza virus (n = 28), and adenovirus (n = 6), in most cases after only one day of culture. For RSV, the sensitivity of the shell vial method was too low (74%) to allow omission of DIF (sensitivity 95%). For the other viruses, the “shell vial/monoclonal antibody mixture” approach was very attractive, being rapid, very specific (⩾97%), and also very sensitive (probably >95%). © 1992 Wiley‐Liss, Inc.
Nested polymerase chain reaction for detection of human immunodeficiency virus type 1 DNA in clinical specimens
Authors: Zazzi Maurizio , Romano Laura , Brasini Agostina , Valensin Pier Egisto
Published: March 2007

A highly sensitive two‐step polymerase chain reaction (PCR) method was evaluated for detection of human immunodeficiency virus type 1 (HIV‐1) DNA in clinical specimens. The product resulting from the first amplification reaction is used as the template for the second PCR with an internal (nested) primer pair. Even when starting from a single copy of HIV‐1 DNA, the double PCR product was readily detected by direct visualization in ethidium bromide‐stained agarose gels. Amplification of minute amounts of HIV‐1 DNA was successful in a considerable excess of HIV‐1 negative DNA than reported previously. All of 85 HIV‐1‐infected individuals were PCR‐positive with at least two of the three sets of primers used, 252 of 255 amplifications allowing unambiguous visualization of a unique DNA fragment of the expected size. The two‐step amplification protocol is simple and rapid and fulfills the requirements of sensitivity and specificity for use in a clinical laboratory. © 1992 Wiley‐Liss, Inc.
Detection of herpesvirus DNA in the large intestine of patients with ulcerative colitis and Crohn's disease using the nested polymerase chain reaction
Authors: Wakefield A. J. , Fox J. D. , Sawyerr A. M. , Taylor J. E. , Sweenie C. H. , Smith M. , Emery V. C. , Hudson M. , Tedder R. S. , Pounder R. E.
Published: March 2007

The prevalence of herpesvirus DNA was examined in inflammatory bowel disease tissue. DNA was extracted from resection and biopsy specimens of the large intestine from patients with ulcerative colitis (n = 21), patients with Crohn's disease (n = 29), and patients with noninflammatory bowel disease (controls) (n = 21). The nested polymerase chain reaction was used to detect viral DNA using primer pairs specific for either cytomegalovirus (CMV), herpes simplex virus 1 (HSV1), human herpesvirus 6 (HHVG), varicella zoster virus (VZV), or Epstein Barr virus (EBV). HSV1 and VZV DNA were not detected in any of the tissue samples. There was a high prevalence of CMV (81%), HHVG (76%), and EBV (76%) DNA in ulcerative colitis tissue compared to Crohn's disease tissues (CMV 66%, HHVG 45%, EBV 55%). Control tissue had a relatively low frequency of CMV (29%) and EBV (19%) DNA but a prevalence of HHVG DNA similar to that of ulcerative colitis (86%). However, the simultaneous presence of HHVG and CMV and/or EBV DNA in ulcerative colitis tissue (76%) was much greater than in either Crohn's disease tissues (38%) or control tissue (29%) (P < 0.05). There was a low prevalence of CMV, HHV6, and EBV DNA in peripheral blood mononuclear cells from all patient groups. CMV and EBV are capable of reactivating HHV6: the high prevalence of coexistent HHV6 infection with either or both of these two viruses in ulcerative colitis tissue suggests that they may play a synergistic role in the pathogenesis of this disease. © 1992 Wiley‐Liss, Inc.
Persistent delta antigenaemia in chronic delta hepatitis and its relation with human immunodeficiency virus infection
Authors: Roingeard Philippe , Dubois Frédéric , Marcellin Patrick , Bernuau Jacques , Bonduelle Sylvie , Benhamou Jean‐Pierre , Goudeau Alain
Published: March 2007

The prevalence of persistent hepatitis delta (HD) antigenaemia and associated factors in patients with chronic infection with the hepatitis delta virus (HDV) were investigated. Among 157 consecutive patients known to be carriers of hepatitis B surface antigen (HBsAg), 36 (23%) had one serum marker of HDV infection (anti‐HD and/or HDAg). Nine of the patients with an HDV marker were HDAg positive, including three who were anti‐HD negative. A follow‐up over a mean period of 13 months showed that five of five patients had a persistent HD antigenaemia. This serological profile was associated with the presence of antibody to the human immunodeficiency virus (anti‐HIV) (P < 0.01), serum HIV antigen (HlVAg) (P < 0.21, and the female sex (P < 0.05). Persistent HD antigenaemia could be the consequence of the suppression of T cell cytotoxic activity against hepatocytes expressing HDAg, a lower humoral response, and/or hormonal factors. © 1992 Wiley‐Liss, Inc.
Serum hepatitis C virus (HCV)‐RNA and response to alpha‐interferon in anti‐HCV positive chronic hepatitis
Authors: Magrin S. , Craxi A. , Fabiano C. , Fiorentino G. , Marino L. , Almasio P. , Pinzello G. B. , Palazzo U. , Vitale M. , Maggio A. , Bucca G. , Gianguzza F. , Shyamala V. , Han J. H. , Pagliaro L.
Published: March 2007

Hepatitis C virus (HCV) replication was assessed before and during alpha‐interferon (IFN) treatment in 22 anti‐HCV positive patients with posttransfusion or sporadic chronic hepatitis (CH). Eleven patients were “responders” and 11 patients “non‐responders” to IFN. Thirteen anti‐HCV negative healthy subjects and five anti‐HCV negative patients with autoimmune CH served as controls.

Serum HCV‐RNA was detected by the polymerase chain reaction (PCR) in all untreated anti‐HCV positive patients but in none of the anti‐HCV negative subjects. PCR primers from the 5′‐non‐coding (NC) region were more sensitive than primers from a non‐structural (NS5) region in detecting HCV‐RNA (21/22, 95% vs. 7/22, 32%, respectively). Positive strand HCV‐RNA titre and positivity rate for the negative strand were similar in responders and non‐responders before IFN treatment, as well as anti‐cl00‐3 titre by enzyme‐linked immunosorbent assay (ELISA), and anti‐5‐1‐1, anti‐c33c, anti‐c22 positivity rate by immunoblot assay (RIBA). HCV‐RNA positivity by both NC and NS primers was more frequent before IFN among responders. During IFN treatment, serum HCV‐RNA was detectable, mostly at low titres, in 1 (NC positive) of the 11 responders and in 9 (4 NS positive and 5 NC positive) of the 11 non‐responders. Among the four non‐responders who were NS positive during IFN, three were NC positive before IFN.

Serum HCV‐RNA was always found in our post‐transfusion or sporadic anti‐HCV positive patients with CH. Viraemia generally decreased during IFN treatment, but no available HCV markers clearly distinguished responders from non‐responders before IFN treatment. Different NC and NS primers behaviour between responders and non‐responders before and during treatment with IFN suggests genomic heterogeneity and may explain non‐responsiveness to IFN. © 1992 Wiley‐Liss, Inc.
Efficient and reproducible new semimicromethod for the detection and titration of HIV in human plasma
Authors: Andreoni M. , Sarmati L. , Parisi S. G. , Ercoli L. , Rocchi G.
Published: March 2007

A semimicromethod was established for isolating human immunodeficiency virus (HIV) in plasma using 48‐well plates and a pool of peripheral blood mononuclear cells (PBMC) from several donors as targets for infection, which increases the efficiency of isolation by reducing the effect of variability due to diverse donor cell susceptibility to HIV infection.

The addition of H9 cells to the PBMC cultures did not affect measurable titers. Nevertheless, it potentiated strongly virus replication in terms of p24 production in the supernatant of the wells with HIV isolates, thus facilitating interpretation of the results. The titration of a virus strain of a known titre and reverse transcriptase activity in parallel provided a constant parameter of efficiency and reproducibility within each experiment, permitting comparison with results from other laboratories.

The reproducibility of the method was highly significant (r = 0.97, P < 0.001); 68% of the 22 plasma samples from HIV‐infected individuals tested by this method were positive. The presence of plasma HIV titer correlated well (P < 0.02) with the low count of CD4+ cells of less than 300/mm3, but not with the presence of the p24 antigen in the serum. © 1992 Wiley‐Liss, Inc.
Risk factors inducing the persistence of high‐risk genital papillomaviruses in the normal cervix
Authors: Vandenvelde C. , Van Beers D.
Published: March 2007

To evaluate the risk factors associated with persistence of human papillomaviruses (HPV) types 16, 18, and 33 in the normal cervix, a prospective study was carried out in Belgium of 323 women without cytological evidence of cervical intraepithelial neoplasia. Demographic and clinical data were obtained by interview, and HPV DNA was assayed in cervical‐swab specimens using the Fast Multiplex Polymerase Chain Reactionbased screening and confirmatory tests. A multivariable linear regression model was constructed using four well‐known risk factors: the use of an oral contraceptive which is either triphasic, or monophasic and containing ethynylestradiol in association with either norethysterone, or levonorgestrel, or lynestrenol, or gestoden, or estrogenic and containing estriol (P = 8 × 10−6) a positive history of genital herpes simplex virus (HSV) infection (P = l0−4), an age inferior or equal to 30 years (P = 0.012), and cigarette smoking (P = 0.020). Crude and adjusted relative risks were calculated for each HPV persistence predictor. The data and the results of the molecular biology of high‐risk genital HPVs are consistent with the hypothesis that the use of an oral contraceptive containing simultaneously and continuously both a potent estrogen and a high activity progestative is necessary to enhance significantly HPV transcription. These observations are also consistent with the hypothesis that the oral contraceptives and HSV genital infection are responsible for HPV persistence in the normal cervix but not for HPV‐induced cervical transformation. The relatively weak correlation found for cigarette smoking may be explained by the very moderate induction effect of phorbol esters on HPV expression, in comparison with their important transforming effect on virus‐infected cells, and thus by the limited number of HPV‐infected women in our study. The fourth predictor probably represents several other risk factors associated with young age such as a high number of sexual partners. © 1992 Wiley‐Liss, Inc.
Hepatitis C virus antibody detection by a line ImmunoAssay and (near) full length genomic RNA detection by a new RNA‐capture polymerase chain reaction
Authors: van Doom Leen‐Jan , van Belkum Alex , Maertens Geert , Quint Wim , Kos Ton , Schellekens Huub
Published: March 2007

A rapid and simple RNA‐capture polymerase chain reaction assay (RCPA) for detection of hepatitis C virus (HCV) is described. The assay detects specifically the presence of (near) full length genomic RNA of HCV by capturing HCV‐RNA at the 3′ terminal end on magnetic beads, followed by cDNA synthesis and PCR with 5′ end specific primers.

Sera were obtained from 30 chimpanzees inoculated with non‐A, non‐B hepatitis material from various sources, 28‐122 months after infection. The sera were tested for the presence of HCV‐RNA by RCPA and for HCV antibodies by a Line ImmunoAssay (Inno‐LIA HCV Ab). Both tests were compared and show a high degree of agreement. Screening of 30 chimpanzee sera revealed either clearing of the virus below detection level (22130) or development of a HCV carrier state (8/30). Only 1 of 11 LIA‐indeterminate samples was positive by RCPA. As the RCPA is more sensitive, it can be used to test for the presence of HCV in sera which are classified indeterminate by the LIA. The outcome of the infection seems to be independent of the nature of the inocula, suggesting that the individual immune response could determine either clearing of the virus or the development of chronic infect ion. © 1992 Wiley‐Liss, Inc.
Controlled trial of prophylactic versus therapeutic use of ganciclovir after liver transplantation in adults
Authors: Cohen Alexander T. , O'Grady John G. , Sutherland Sheena , Sallie Richard , Tan K.‐C. , Williams Roger
Published: March 2007

The potential of prophylactic ganciclovir for the control of cytomegalovirus (CMV) infection was evaluated in a prospective controlled trial of 65 patients after liver transplantation. A group of 33 patients received ganciclovir (10 mg/kg/day) during the third and fourth weeks after transplantation, while 32 patients were randomised to receive ganciclovir (10 mg/kg/day) only when clinical CMV disease was diagnosed. Eight patients (25%) in the latter group received ganciclovir and included the only death attributed to CMV infection in the study. Prophylactic ganciclovir was associated with a lower incidence of serologically diagnosed secondary infection and the development of the IgM anti‐CMV antibody. However, the frequency of clinical infections was similar in the two groups (9/33 vs. 11/32). Liver function tests during the second month after transplantation were not significantly better in the patients receiving prophylactic ganciclovir. Leucopaenia was not seen in patients receiving prophylactic ganciclovir. © 1993 Wiley‐Liss, Inc.
Hepatitis A antibody titres after infection and immunization: implications for passive and active immunization
Authors: Zaaijer H. L. , Leentvaar‐Kuijpers A. , Rotman H. , Lelie P. N.
Published: March 2007

Titres of antibodies against hepatitis A virus (HAV) were determined in patients, in donors, and in volunteers after active, passive, and combined immunization. Highest titres were found in recently infected persons: in 109 IgM anti‐HAV positive persons, the geometric mean titre (GMT) was 15,400 mlU/ml. The GMT in 265 anti‐HAV positive blood donors was 10,700 mlU/ml. The anti‐HAV seroprevalence in 19,746 donors increases with age: at the age of 40 years, 50% have antibodies. Titres after active, passive, and combined immunization were studied in three groups: 51 persons received inactivated HAV vaccine at months 0, 1, and 6. The GMT after the booster was 3,400 mlU/ml at month 7. All persons produced more than 100 mlU/ml anti‐HAV. Forty‐nine persons received both 5 ml immunoglobulin and three vaccinations, yielding a GMT of 1,300 mlU/ml at month 7. One person in this group produced less than 100 mlU/ml anti‐HAV. Forty‐nine persons received 5 ml immunoglobulin intramuscularly. At day 5 the GMT was 96 mlU/ml. The estimated minimum protective level (10 mlU/ml) was reached in 3 months. Hepatitis A vaccination may supersede the use of immunoglobulin as prophylaxis for travellers to endemic areas. Passive immunization remains necessary for protection during outbreaks. The dosage regimen for passive immunization is based on old studies using preparations with unknown anti‐HAV content. Concern regarding the antibody levels in immunoglobulin preparations is justified; the prevalence of HAV antibodies in developed countries continues to fall. Our results indicate that dosage regimens should be reconsidered. Dosage should be deduced logically from the anti‐HAV antibody content of the immunoglobulin preparation. © 1993 Wiley‐Liss, Inc.
Three cases of human herpesvirus‐6 latent infection: Integration of viral genome in peripheral blood mononuclear cell DNA
Authors: Luppi Mario , Marasca Roberto , Barozzi Patrizia , Ferrari Sergio , Ceccherini‐Nelli Luca , Batoni Giovanna , Merelli Elisa , Torelli Giuseppe
Published: March 2007

Saliva and peripheral blood mononuclear cells from three patients, two with lymphoproliferative disorders and one suffering from multiple sclerosis, were examined for the presence of human herpesvirus‐6 (HHV‐6) genome by using the polymerase chain reaction and Southern blot analysis. The search for anti‐HHV‐6 antibodies, carried out in the sera of the same cases by an immunofluorescence assay, was negative in two cases at the lowest dilution used (1:40). These three patients had a high number of HHV‐6 specific sequences in uncultured peripheral blood mononuclear cells, which are thought to be a normal site of viral latency although, in healthy individuals, the infected cells are extremely rare. In order to gain some insight into the state of the viral genome in this latent HHV‐6 infection, we used pulsed field gel electrophoresis to separate HHV‐6 DNA directly from HHV‐6 (strain GS) infected HSB‐2 cells and from the peripheral blood mononuclear cells of these three patients. Our study showed the presence of intact viral genome, of the expected length of 170 kb, persisting as free extrachromosomal element in the HSB‐2 cells but not in patients' peripheral blood mononuclear cells. On the other hand, in strong contrast with the results obtained in infected HSB‐2 DNA, the restriction analysis of the three patients' peripheral blood mononuclear cell DNA showed fragments of molecular weight constantly higher than the 170 kb segment, indicating that the viral sequences are linked to high molecular weight cellular DNA. Our findings are consistent only with a latent infection in which HHV‐6 is integrated in vivo and suggest that pulsed field gel electrophoresis analysis is well worth using to evaluate the presence of integrated, intact, or fragmented viral genomes in HHV‐6 associated lymphoproliferative diseases and immune disorders. © 1993 Wiley‐Liss, Inc.
Antibody response to nonstructural proteins of hepatitis A virus following infection
Authors: Robertson Betty H. , Jia Xi‐Yu , Tian Houwen , Margolis Harold S. , Summers Donald F. , Ehrenfeld Ellie
Published: March 2007

The nonstructural proteins of hepatitis A virus (HAV), produced during active virus replication, are alternative antigens that could be used to differentiate disease from inactivated vaccine‐induced antibodies. An assay based on immune precipitation of proteins translated from transcripts of the P2 region of viral cDNA was used to evaluate the development of antibodies after natural infection or vaccination. Antibodies against P2 proteins were found in all sera from clinical cases of hepatitis A following the acute phase. Chimpanzees vaccinated with inactivated or cell‐adapted HAV had no detectable antibodies against P2 products, either before or after wild type virus challenge. A serosurvey of sera positive for total anti‐HAV (HAVAB, Abbott Laboratories, North Chicago) suggested that some individuals had no detectable antibodies to the P2 antigen by immune precipitation. These results were attributed to the lower sensitivity of the immunoprecipitation assay, since antibodies to capsid proteins, as measured by immunoprecipitation, were also not detected in most of these sera. © 1993 Wiley‐Liss, Inc.
Is the course of perinatal hepatitis B virus infection influenced by genetic heterogeneity of the virus?
Authors: Raimondo Giovanni , Tanzi Elisabetta , Brancatelli Santa , Campo Salvatore , Sardo Maria A. , Rodinò Giuseppina , Pernice Maurizio , Zanetti Alessandro R.
Published: March 2007

We studied the relations between genetic heterogeneity of pre‐C region of hepatitis B virus (HBV) DNA and outcome of HBV infection in 5 infants with perinatal infection, 3 born to antihepatitis B e antigen (HBeAg), and 2 to HBeAg positive mothers. HBV infection developed in the babies at 3–4 months of age, but it resolved with seroconversion to anti‐HBs in infants born to anti‐HBe positive mothers, while the infection became chronic in the 2 babies born to HBeAg positive mothers. HBV‐DNA extracted from the first hepatitis B surface antigen (HBsAg) positive serum sample of each baby was amplified and directly sequenced for the pre‐core region. HBV‐DNA sequences from 3 babies born to anti‐HBe positive mothers showed at position 1896 the contemporary presence of 2 nucleotides (G + A), indicating a mixture of wild‐type and "e minus"variant HBV. These findings suggest a possible co‐transmission of the 2 viruses from anti‐HBe positive mothers to newborns. HBV‐DNA from babies born to HBeAg positive mothers showed wild‐type sequences only. The results of this study suggest that the outcome of HBV infection in newborns depends not only on the host's immunocompetence and on viremia level in maternal blood, but also on heterogeneity of HBV. Transmission of mixed HBV populations appears associated with an early immunoelimination of the virus, while infection with wild‐type HBV alone contributes to induction of chronicity. © 1993 Wiley‐Liss, Inc.
Identification and molecular characterization of two diagnostically relevant marker proteins of the Epstein‐Barr virus capsid antigen complex
Authors: Van Grunsven Wout M. J. , Nabbe Arjan , Middeldorp Jaap M.
Published: March 2007

The molecular specificity of the IgG response against Epstein‐Barr virus (EBV) was studied in 345 randomly collected sera of normal healthy individuals. The sera were tested on immunoblots containing antigens of the cell line HH514.c16 (a superinducible derivate of P3HR1), noninduced or induced for the expression of early antigens (EA) or viral capsid antigens (VCA), and from the EBV‐negative cell line Ramos‐Nut. This study reveals a remarkable similar antigen recognition pattern of IgG class antibodies in sera of healthy EBV carriers. The protein bands recognized predominantly have molecular weights of 18 kD, 36/38 kD, 40 kD, 72 kD, and 160 kD. The 72 kD and 36/38 kD bands were identified as EBNA1 and "Zebra," respectively, using reading frame‐specific antisera. The bands at 160 kD (major capsid protein), 40 kD, and 18 kD were identified as VCA‐class proteins. Of all EBV‐seropositive sera tested, 98% reacted with either p18 or p40 or both. The synthesis of the antigens p18 and p40 was inhibited by phosphonoacetic acid, indicating that these were true late proteins. The detection of p18 and p40 in purified virion and capsid preparations confirms that these proteins are structural components of viral capsid antigen complex. © 1993 Wiley‐Liss, Inc.
Variability of IgM response in hepatitis C virus infection
Authors: Zaaijer H. L. , Mimms L. T. , Cuypers H. T. M. , Reesink H. W. , Van Der Poel C. L. , Taskar S. , Lelie P. N.
Published: March 2007

The IgM and IgG antibody response to various hepatitis C virus (HCV) antigens was studied in 8 patients who acquired posttransfusion HCV infection. IgM anti‐HCV was detectable in only 4 of these patients, coincident with (1 patient) or later than (3 patients) the IgG anti‐HCV response. Seven patients had initially decreasing IgG anti‐HCV titres, indicating passive transfer of antibodies from donor to recipient. All 8 patients showed active IgG seroconversion, as demonstrated by increasing IgG anti‐HCV titres, on average, 75 days after infection. Five years after infection, all patients were still reactive for IgG anti‐HCV antibodies and 7 were positive for HCV RNA by the polymerase chain reaction (PCR). Two of these PCR positive patients were also reactive for IgM anti‐HCV. It is concluded that the serology of HCV infection does not follow the classical pattern of IgM response preceding detection of IgG. The IgM response may be absent, late, or persistent after HCV infection. The serological diagnosis of recent HCV infection should be based on the polymerase chain reaction or rising IgG titres in at least 2 sequential patient blood samples. © 1993 Wiley‐Liss, Inc.
Serological studies of an enterically transmitted non‐A, non‐B hepatitis in Somalia
Authors: Mushahwar Isa K. , Dawson George J. , Bile Khalif M. , Magnius Lars O.
Published: March 2007

An outbreak of an acute sporadic viral hepatitis occurring at three villages in the lower Shebli region of southern Somalia was studied. Sera were examined for antibodies to hepatitis E virus (HEV) antigen by a recently developed enzymelinked immunosorbent assay. The prevalence of anti‐HEV ranged from 77.8% to 94.0% among the three villages. Anti‐HEV prevalence did not differ significantly with age or sex. HEV infections were widely distributed among all age groups, without a preponderance in any specific age category. Overall, 97.6% of afflicted patients who were examined 6–10 days after onset of jaundice developed either IgM or IgG anti‐HEV. © 1993 Wiley‐Liss, Inc.
Absence of human papillomavirus type 16 E6 transcripts in HPV 16‐infected, cytologically normal cervical scrapings
Authors: Falcinelli Cristina , Van Belkum Alex , Schrauwen Lianne , Seldenrijk Kees , Quint Wim G. V.
Published: March 2007

By a combination of reversed transcription and subsequent polymerase chain reaction (RNA‐PCR), 23 cytologically normal cervical scrapings, positive for the presence of human papillomavirus type 16 (HPV 16) DNA, were analyzed for the presence of transcripts originating from the E6 region of the viral genome. This region is thought to be involved in transformational, tumorigenic events. No mRNAs of the E6 region were detectable using the most sensitive PCR‐mediated procedure currently available. Since it was previously shown that in cytological abnormal cervical scrapings about one‐half of the samples positive for HPV 16 DNA express mRNAs of the E6 region, a difference between normal and abnormal cervical scrapings, when the HPV 16 is present, exists. The observed difference between cytologically normal and abnormal, HPV‐DNA‐positive cervical scrapes may eventually be used as a prognostic marker for screening of women at risk for the development of cervical carcinoma. However, firm establishment of the putative correlation between tumor progression and the presence of E6 transcripts requires extensive follow‐up analysis of HPV‐positive patients. © 1993 Wiley‐Liss, Inc.
Hepatitis C virus infection in hemodialysis patients in southern Sweden: Epidemiological, clinical, and diagnostic aspects
Authors: Nordenfelt E. , Löfgren B. , Widell A. , Hansson B.‐G. , Zhang Y. Y. , Hagstam K.‐E. , Kurkus J.
Published: March 2007

A prevalence of hepatitis C virus (HCV) antibodies of 12% was found in 276 patients from 11 dialysis units. Between zero and 22% of the patients in the different units were anti‐HCV positive. The epidemiology of HCV was studied in two units during a 2 year period by antibody assays and the polymerase chain reaction and correlated with clinical manifestations. Two types of epidemiologic patterns were found that may explain the wide difference of HCV prevalence described in different dialysis units. In one unit there was no evidence of spread within the unit, and the prevalence of HCV was dependent on the status of the patients entering for treatment. In the other unit, a clustering of infected patients could be seen in which 13 of 36 were infected during a 3 year period. Some patients who had not received blood transfusions were among the infected. Hepatitis C infection was the most common explanation for repeated abnormal transferase levels. Most of the HCV‐infected patients reacted both for anti‐HCV and HCV RNA. HCV RNA was in general detected earlier than anti‐HCV seroconversion. Among 20 HCV RNApositive serum samples that were anti‐HCV ELISA‐positive 18 had indeterminate and two negative reactions by immunoblot (RIBA 2). Thus the RIBA 2 test should be used with caution as a confirmatory antibody test in this group of patients. © 1993 Wiley‐Liss, Inc.
Epstein‐Barr virus genomic sequences and specific antibodies in cerebrospinal fluid in children with neurologic complications of acute and reactivated EBV infections
Authors: Imai Shosuke , Usui Norio , Sugiura Makoto , Osato Toyoro , Sato Toshiya , Tsutsumi Hiroyuki , Tachi Nobutada , Nakata Shuji , Yamanaka Tatsuru , Chiba Shunzo , Shimada Masamitsu
Published: March 2007

Four children with infectious mononucleosis (IM) and one with reactivated Epstein‐Barr virus (EBV) infection had concomitant central nervous system disorders. Cerebrospinal fluid (CSF) samples from all five patients contained EBV genomic sequences and EBV‐specific antibodies in the neurologic stage, but not during convalescence. Cerebrospinal fluid from two non‐neurologic IM patients had neither EBV DNA nor EBV antibodies. The EBV‐positive CSF of the five with neurological disorders were aseptic in culture and all negative for other human herpesvirus DNAs and antibodies: herpes simplex virus types 1 and 2, cytomegalovirus, varicella‐zoster virus, and human herpesvirus 6. Epstein‐Barr virus DNA and EBV antibodies were not detected in the CSF of 17 EBV‐seropositive patients with mumps meningitis, rubella encephalitis, unknown febrile convulsion, or partial epilepsy. It is suggested that EBV plays a causal role in neurologic manifestations in patients with acute and reactivated EBV infections, through direct viral invasion and immunopathological reactions. © 1993 Wiley‐Liss, Inc.
Quantitative assessment of hepatitis B virus DNA in chronic hepatitis B: comparison of two solution hybridization assays
Authors: Janssen Harry L. A. , Schoenmaker‐Weber Yvonne A. M. , Kruining Hans , Schalm Solko W. , Heijtink Rudolf A.
Published: March 2007

We compared two solution hybridization assays, the AffiProbe assay (Orion Corporation) and the Abbott HBV‐DNA assay (Abbott), for quantitative measurement of hepatitis B virus (HBV) DNA in serum samples obtained from chronic hepatitis B surface antigen (HBsAg) carriers. Forty transversally collected (group 1) and 83 serially collected (group 2) serum samples from chronic hepatitis B patients were tested with both assays. The serial serum samples were obtained from 6 patients who underwent α‐interferon therapy with different outcomes (nonresponse, hepatitis B e antigen [HBeAg] seroconversion, HBeAg and HBsAg seroconversion). In group 1 a good correlation (r = 0.91; P < 0.001) was found between the HBV‐DNA results of the two assays. Two samples (5%) were HBV‐DNA positive according to the Abbott but negative according to the AffiProbe assay; for all other samples the HBV‐DNA status corresponded. In group 2 the assays gave colinear HBV‐DNA results during follow‐up of 5 of the 6 patients (correlation for the total group: r = 0.90; P < 0.001). Nevertheless, in both groups the AffiProbe assay yielded about 5–10 times higher HBV‐DNA levels than the Abbott HBV‐DNA assay (P < 0.001). These discordant results were most probably due to standardization differences of the positive control samples of the two test systems. This observation underlines the need for international standardization of HBV‐DNA and uniform reference panels. © 1993 Wiley‐Liss, Inc.
Preliminary evidence that a trpE‐HEV fusion protein protects cynomolgus macaques against challenge with wild‐type hepatitis e virus (HEV)
Authors: Purdy Michael A. , McCaustland Karen A. , Krawczynski Kris , Spelbring John , Reyes Gregory R. , Bradley Daniel W.
Published: March 2007

Immunization of two cynomolgus macaques (cynos) with trpE‐C2 protein, a trpE‐HEV fusion protein that represents the carboxyl two thirds of the putative capsid protein, prevented development of biochemical evidence of viral hepatitis in these primates after challenge by wild‐type HEV from either a Burmese or Mexican stool isolate. Neither of the immunized animals showed any elevation of alanine aminotransferase activity after challenge with wild‐type HEV in marked contrast with the unimmunized (control) cynos. In the case of the Burmese HEV challenged cyno, the protective effect was complete with the animal failing to demonstrate any evidence of HEV infection. The immunized cyno challenged with Burmese HEV did not exhibit any HEV RNA in its stools or HEV antigen in its liver. The immunized cyno (#8902) challenged with Mexican virus exhibited HEV RNA in its stools and HEV antigen in its liver; however, microscopic examination of liver biopsy specimens from this cyno failed to detect histopathologic evidence of viral hepatitis. All of the animals (naive and immunized) developed anti‐HEV IgM and IgG responses after HEV challenge. Our preliminary studies indicate that the trpE‐C2 protein is a promising candidate HEV vaccine.
Accelerated schedule of hepatitis B vaccination in patients with hemophilia
Authors: Santagostino Elena , Mannucci Pier Mannuccio , Gringeri Alessandro , Rumi Maria Grazia , Rafanelli Daniela , Rocino Angiola , Schiavoni Mario , Chistolini Antonio , Bona Eros Di , Muleo Gaetano
Published: March 2007

Early development of immunity after hepatitis B vaccination is particularly important for patients such as hemophiliacs, at high risk for acquiring hepatitis B from potentially infectious plasmaderived concentrates. The purpose of this study was to evaluate whether or not protective antibody titers could be achieved quickly and maintained in hemophiliacs by an accelerated vaccination schedule. A yeast‐recombinant hepatitis B vaccine (Engerix B, SKF Ritt) was given subcutaneously in the deltoid region and repeated 2 and 6 weeks later to 85 hemophiliacs negative for hepatitis B virus (HBV) markers. After the first 22 patients had been enrolled, a modification of the schedule involving a fourth booster dose 24 weeks after the first dose of vaccine was applied to the next 63 consecutive vaccinees.

Fifty‐three percent of vaccinees had antibody titers to hepatitis B surface antigen (anti‐HBs≥ 10 mlU/ml) by week 6, even though the mean titers of anti‐HBs were somewhat lower than those achieved historically in normal individuals. The protection rate had increased to 87% by week 10, one month after the third dose of vaccine, and to 93% by week 24. One year after starting vaccination, the rate for the vaccinees who did not receive the fourth booster dose was 71%, and 96% for those who did receive the fourth dose, with only 2 patients not responding despite the booster dose. It is concluded that even though the accelerated schedule of immunization produced rapidly high rates of protective antibody titers, a booster dose is required to obtain higher titers and provide more persistent immunity.
Hepatitis C viral RNA titers in serum prior to, during, and after oral treatment with ribavirin for chronic hepatitis C
Authors: Reichard Olle , Yun Zhi‐Bing , Sönnerborg Anders , Weiland Ola
Published: March 2007

Ten patients with biopsy verified chronic hepatitis C virus (HCV) infection were treated with oral ribavirin at a dose of 1,000–1,200 mg per day in two divided doses for 12 weeks. Serum alanine aminotransferase (ALT) levels and hepatitis C viral ribonucleic acid (RNA) levels in serum were followed prior to, during, and 12 weeks posttreatment. ALT levels decreased significantly in all patients during therapy from a mean level of 3.21 μkat/l (range 1.22 to 7.79) before, to 1.25 μkat/l (range 0.78 to 2.04) at the end of treatment (P < 0.005). Hereafter, relapse to pretreatment levels was seen within 12 weeks after treatment stop. The hepatitis C viral RNA levels decreased from a mean 10 log titer of 4.1 (range 1–6) before treatment to 3.4 (range 1–5) at treatment stop. Five patients did not change their HCV RNA titers during treatment. Twelve weeks posttreatment only 3 patients had lower titers than prior to treatment. We conclude that oral ribavirin seems to reduce the viral load, at least temporarily, in some patients with chronic viremic HCV infection. Further studies are needed to evaluate fully the effect of oral ribavirin on chronic HCV infection.
Specific deletions in the hepatitis B virus core open reading frame in patients with chronic active hepatitis B
Authors: Ackrill Andrew M. , Naoumov Nikolai V. , Eddleston Adrian L. W. F. , Williams Roger
Published: March 2007

The polymerase chain reaction (PCR) and DNA sequencing were used to examine genomic variation in the pre‐core/core open reading frame of the hepatitis B virus (HBV) in chronically infected patients. Gel electrophoresis of amplification products showed the presence of shortened forms of the core gene in addition to the full length product. These shortened forms were seen only in patients with chronic active hepatitis (CAH) seropositive for HBeAg and not in patients with chronic persistent hepatitis (CPH) or HBeAg minus CAH. Cloning and DNA sequencing revealed the presence of a number of overlapping deletions within the core gene, the majority being in‐frame, which were clustered within aa 81‐114 of the core gene product. These deletions were found in patients with CAH from different racial and geographical backgrounds, whereas PCR analysis of the surface and ×open reading frames showed no shortened forms suggesting deletions to be specific to the core gene in these patients. Because the product of the core gene‐the HBV core antigen–is believed to be the major target for T‐cell‐mediated liver damage, it seems likely that the products of core genes carrying deletions will alter immune recognition and may be of importance in the progression of inflammatory liver damage.
Efficacy of a recombinant Norwalk virus protein enzyme immunoassay for the diagnosis of infections with Norwalk virus and other human “candidate” caliciviruses
Authors: Parker Simon , Cubitt David , Jiang Jason Xi , Estes Mary
Published: March 2007

A recombinant Norwalk virus antigen enzyme immunoassay (rNV EIA) for the detection of Norwalk virus (NV) antibodies was evaluated using sera from volunteers and patients infected with Norwalk, Hawaii, Snow Mountain Agent, small round structured viruses (SRSV), and human calicivirus (HuCV) strains. The results for NV antibodies correlated well with previous results obtained using a NV radioimmunoassay. The test rNV EIA was found to be highly specific for NV. The results of tests on sera obtained from patients involved in confirmed outbreaks of SRSV in the UK showed that only 1/12 was due to Norwalk virus. Further evidence is presented that SRSV strain UK2 is antigenically related to NV.
Partial control of hepatitis delta virus superinfection by immunisation of woodchucks (Marmota monax) with hepatitis delta antigen expressed by a recombinant vaccinia or baculovirus
Authors: Karayiannis P. , Saldanha J. , Jackson A. M. , Luther S. , Goldin R. , Monjardino J. , Thomas H. C.
Published: March 2007

We have successfully limited the level of hepatitis delta viraemia occurring after superinfection of hepadna‐virus infected woodchucks by prior immunisation with the short form of the hepatitis delta virus antigen expressed by a recombinant baculovirus or vaccinia virus. This phenomenon occurred in the absence of detectable circulating antibody to hepatitis delta virus antigen and in the absence of evidence of priming of the humoral immune response and may reflect the induction of a cytotoxic T‐cell response. The latter would control viraemia by rapid lysis of delta antigen expressing hepatocytes. It is suggested that the T‐cell epitopes involved may be located on the carboxyl end of the delta protein (amino acids 77‐195).
Efficacy of (S)‐1‐(3‐hydroxy‐2‐phosphonylmethoxypropyl)cytosine for the treatment of lethal vaccinia virus infections in severe combined immune deficiency (SCID) mice
Authors: Neyts Johan , Clercq Erik De
Published: March 2007

Severe combined immune deficient (SCID) mice inoculated intravenously with vaccinia virus (VV) became sick within 6–8 days and died 10–12 days after infection. Tail lesions developed and the number depended on the virus inoculum. Age‐matched immunocompetent NMRI mice similarly infected also developed tail lesions but did not become sick. When the infected SCID mice were treated with the acyclic nucleoside phosphonate HPMPC [(S)‐1‐(3‐hydroxy‐2‐phosphonylmethoxypropyl) cytosine], either for 5 consecutive days starting on the day of infection or for 5 consecutive days starting on day 2, 4, or 6 post infection, or as a single dose at 7 days or 1 day before infection, VV‐associated death was significantly delayed. VV‐infected SCID mice that received two doses of 20 mg/kg of HPMPC every week survived the infection for about 130 days. The period during which the mice remained disease‐free following HPMPC treatment correlated with the absence of detectable virus in their organs. The VV/SCID mouse model employed here may be useful for determining whether (attenuated) recombinant VV (carrying HIV genes) may have detrimental effects in the immunodeficient host. HPMPC may be considered as a drug candidate for the treatment and prophylaxis of such complications.