bluuebunny/crossref_metadata_2025 huggingface dataset:
Hepatitis B virus X‐gene product: A promiscuous transcriptional activator
Authors: Rossner Michael T.
Published: March 2007

The disease state induced by infection with hepatitis B virus (HBV) is manifest in varying ways characterised by the extent of liver inflammation and damage and viral persistence. In a small percentage of cases, primary infection leads to fulminant hepatitis resulting in severe liver dysfunction with very high mortality. Primary infection is most often resolved by complete clearance of the virus and development of immune memory to counter reinfection, but 5‐10% of infected adults develop chronic infection characterised by the persistence of viral antigens in the serum and accompanied by varying degrees of hepatic injury. This disease state may continue after integration of HBV DNA into the hepatocyte genome from which transcription of viral antigen genes may continue in the absence of virion production. Chronically infected patients are predisposed to developing hepatocellular carcinoma (HCC) [Szmuness, 1978] with more than 100‐fold greater probability than non‐infected individuals [Beasley, et al., 1981]. Hepatitis B constitutes a major worldwide health problem with the number of chronically infected people currently estimated in excess of 250 million.

HBV is the prototype member of the family of hepadnaviridae, which includes hepatitis viruses isolated from the woodchuck (WHV) [Summers et al., 1978], ground squirrel (GSHV) [Marion et al., 1980], Pekin duck (DHBV) [Mason et al., 1980], and grey heron (HHV) [Sprengel et al., 1988]. This taxonomy is derived from the relative hepatropism of virus family members, their common virion morphology, genome size, structure and organisation, and common mechanism of genome replication, which proceeds through reverse transcription of an RNA intermediate [Summers and Mason, 1982] in a manner analogous to that of retroviruses. All the viruses exhibit a strict host specificity, the human virus replicating only in man and a small number of higher apes.

HBV has a partially double stranded, open‐circular genome of 3.2 kilobases (kb) which contains four open reading frames (ORFs) including the S‐gene encoding the surface antigen and the C‐gene encoding the core antigen (Fig. 1 ). A large ORF encompassing most of the viral genome encodes the viral polymerase while the fourth ORF encodes a protein of 154 amino acid residues which has been termed the X antigen (HBxAg) because the function of this product in the viral lifecycle is still under intensive investigation.
Replication and cytopathaology of human parvovirus B19 in human umbilical cord blood erythroid progenitor cells
Authors: Sosa Carlos E. , Mahony James B. , Luinstra Kathleen E. , Sternbach Marion , Chernesky Max A.
Published: March 2007

Human parvovirus B19 productively infected erythroid progenitor (EP) cells from umbilical cord blood, in vitro as shown by an increase of viral DNA in supernatant fluid assayed by dot blot hybridization and liquid scintillation counting. Progeny virus was released into the supernatant fluid of CD34+ EP cells which had been purified by immunomagnetic separation. This supernatant fluid was infectious for bone marrow cells. Erythroid bursts infected with virus showed characteristic cytopathic effect by electron microscopy consisting of cytoplasmic vacuolization, marginated chromatin, and nuclear inclusions of lattice or crystalline arrays. Cultures of umbilical cord blood EP cells may be useful for the propagation of parvovirus 619 serological testing reagents and the study of virus‐host cell interactions.
Mucosal T cell distribution during infection with respiratory syncytial virus
Authors: Kimpen Jan L. L. , Rich Gary A. , Mohar Claudia K. , Ogra Pearay L.
Published: March 2007

Groups of 12‐week‐old Balb/c mice were inoculated intranasally with respiratory syncytial virus (RSV) and sacrificed at regular intervals after infection. T lymphocyte subset distribution was determined in lung tissue, bronchoalveolar lavage (BAL), peripheral blood, and spleen by means of flow cytometry employing monoclonal antibodies against the T cell membrane antigens Thyl.2 (pan‐T), Ly2 (CD8), and L3T4 (CD4). Thyl.2′ cells increased in the lung from 35.4% of total lymphocytes before infection to 47.6% on day 7 after infection. This increase was largely accounted for by an increase in Ly2+ cells, which manifested a rise from 7.8% preinfection to 19.8% on day 7. The level of L3T4+ cells remained constant (27.9% preinfection vs. 25.2% on day 7). The L3T4+/Ly2+ ratio in the lungs reached a nadir 7 days post infection (1.5 vs. 3.5 before infection). The total cell count in BAL increased more than tenfold during the first week after infection. At the same time Thyl.2+ cells in the BAL increased from 41.1% of total lymphocytes on day 1 to 85.3% on day 7. Ly2+ influx was the most important (5.8% on day 1 vs. 41.1% on day 7). L3T4+ cell levels increased from 17.2% on day 1 to 40.1% on day 7. RSV‐specific lymphocyte transformation was observed in BAL and blood but not in the lung tissue and spleen on day 7 postinfection. The disappearance of infectious virus in the lung correlated directly to the peak appearance of Ly2+ T cells in the lung tissue and BAL.
Differential IgG avidity to rubella virus structural proteins
Authors: Mauracher Christoph A. , Mitchell Leslie Ann , Tingle Aubrey J.
Published: March 2007

Avidity maturation of IgG antibody responses directed against the structural proteins of rubella virus ( E1, E2, and C) as well as whole rubella virus (RV) was assessed at sequential time intervals in 7 individuals following serologically confirmed wild rubella infection. Individual structural proteins were purified from tissue culture supernatants by differential centrifugation, followed by preparative SDS‐PAGE under non‐reducing conditions. Avidity of IgG anti‐rubella responses was measured by using the 8 M urea elution technique and results expressed as an elution ratio [ER(%)]. A low mean ER(%) of 23% was determined for E1‐specific IgG responses during the 10‐20 day period following onset of clinical rubella, with subsequent maturation of avidity ER(%) values to 52%, 75%, and 84% at 3 months, 1 year, and 2 years, respectively, post‐rubella. In contrast, IgG anti‐E2 responses showed minimal avidity maturation with ER(%) values of 20%, 29%, 30%, and 31% over the same time intervals. Similarly, responses to the capsid protein (C) remained at low avidity ER(%) values of 21%, 29%, 36%, and 35% over the 2 year follow‐up period. The avidity maturation values for IgG directed against whole RV preparations paralleled observations for E1 ‐specific responses with ER(%) values of 23%, 52% 85%, and 87%, respectively. These data support the need to assess individual protein‐specific antibody avidities in order to more fully understand viral‐specific immune responses.
Infection enhancement of influenza A NWS virus in primary murine macrophages by anti‐hemagglutinin monoclonal antibody
Authors: Ochiai Hiroshi , Kurokawa Masahiko , Matsui Shoko , Yamamoto Tatsuru , Kuroki Yoshito , Kishimoto Chiharu , Shiraki Kimiyasu
Published: March 2007

Antibody‐dependent enhancement (ADE) of influenza A NWS virus infection was investigated in primary murine macrophages (Mø,) using antihemagglutinin(HA) monoclonal antibody (mAB). Contrary to previous reports of abortive influenza virus infection in primary Mø, this study demonstrated that the NWS virus replicated productively in both resident peritoneal Mø, and thioglycolate‐elicited peritoneal Mø, providing cleavage of the HA was achieved by trypsin; 5 μMg/ml of trypsin was the optimum concentration for the induction of infectivity. Under multiplecycle growth conditions in the presence of mAB at various concentrations in trypsin‐containing media, ADE was demonstrated in both Mø, in the presence of subneutralizing concentrations of mAB. Flow cytometric analysis showed that the mechanism of virus entry into Mø, could be through HA to specific virus receptors, or HA plus antibody to Fc receptors. These results indicate that ADE of the NWS virus infection actually occurs on Fc receptor‐bearing primary murine Mø, depending on the concentration of antibody in the presence of the appropriate protease for cleavage of viral HA.
High rate of multiple genital HPV infections detected by DNA hybridization
Authors: Lauricella‐Lefebvre M.‐A. , Piette J. , Lifrange E. , Lambotte R. , Gerard P. , Rentier B.
Published: March 2007

Cervical smears collected from 450 patients involved in a clinical follow‐up of cervical human papillomaviruses (HPV) infections were screened for the presence of HPV 6b, 11, 16, and 18 DNA by both dot blot and southern blot hybridization methods. Using very high stringency hybridization assays, the four HPV types could be easily distinguished by dot blotting. After a preliminary clinical sorting, 42.9% of the samples were found to be HPV‐positive. Among the samples infected by a single HPV, type 16 was the most frequent (25.4% of the positive samples) followed by 6b (19.7%), 11 (8.3%), and 18 (7.2%). Double or even multiple infections by the different HPV types were detected at a very high rate (39.4% of the positive samples).
HIV‐1 reverse transcriptase inhibiting antibody titer in serum: Relation to disease progression and to core‐antibody levels
Authors: Neumüller Magnus , Karlsson Anders , Lennerstrand Johan , Källander Clas F. R. , Sandstrdm Erik , Holmberg Viveca , Gronowitz J. Simon
Published: March 2007

A new assay for detecting inhibition of reverse transcriptase activity (the RT‐i REA) was developed. This assay was standardized for screening serum samples for reverse transcriptase inhibiting antibodies (RT‐iAb). High specificity (100%) and sensitivity (<98%) were achieved with samples from HIV‐negative individuals and HIV‐in‐fected individuals. The RT‐i REA was also used in a study of the titers of RT‐iAb in serum samples obtained from 33 HIV‐infected homosexual men. The results confirmed the relation between decreasing RT‐iAb levels and progression to late stages of the disease. Furthermore, a falling RT‐ iAb titer was observed in 14 of 15 individuals experiencing periods of severe clinical symptoms attributed to HIV‐activity. In 7 of the patients the decline in RT‐iAb titer began prior to severe clinical symptoms. The fall in RT‐iAb titer also correlated with a reduction in core Ab level. The core Ab level has previously been reported to be a disease progression marker with considerable prognostic value. However, whereas all patients were positive for RT‐iAb, 8 of the 33 patients did not have detectable core Ab. The use of RT‐iAb titer as a marker of disease progression is discussed.
Serologic differentiation of human T lymphotropic virus type I from type II infection by synthetic peptide immunoassays
Authors: Roberts Chester R. , Mitra Rita , Hyams Kenneth , Brodine Stephanie K. , Lal Renu B.
Published: March 2007

Synthetic peptide‐based serologic assays (Select‐HTLV and SynthEIA) that distinguish the closely related human T‐cell lymphotropic virus types I (HTLV‐I) and II (HTLV‐II) were tested blindly for their ability to correctly identify infection caused by either virus type. Of 57 HTLV‐I and 38 HTLV‐II specimens from individuals whose infections were confirmed by polymerase chain reaction, the Select‐HTLV assay categorized 56(98%) as HTLV‐I, 36 (95%) as HTLV‐II, and 3 (3%) as nontypeable. Similarly, the SynthEIA assay categorized 54 (95%) as HTLV‐I, 29 (76%) as HTLV‐II, and 12 (13%) as nontypeable. More importantly, no specimen was wrongly classified by either assay (100% specificity). Further, analysis of serial specimens from six patients also demonstrated concordant results with the PCR findings. Our results suggest that serotyping of HTLV infections can be achieved reliably by these peptide assays, thus the need for technically complex PCR‐based HTLV typing, while not eliminated, can be greatly reduced.
Detection of multiple Epstein‐Barr viral strains in allogeneic bone marrow transplant recipients
Authors: Gratama J. W. , Lennette E. T. , Lönnqvist B. , Oosterveer M. A. P. , Klein G. , Ringdén O. , Ernberg I.
Published: March 2007

We have previously shown in 3 allogeneic bone‐marrow transplant (BMT) recipients that complete replacement of recipient marrow was associated with the elimination of the pretransplant Epstein‐Barr virus (EBV) strain of the recipient. To study the kinetics of EBV elimination and rein‐fection in more detail, we have performed a longitudinal study of BMT recipients combining serology, virus isolation from mouthwashes and peripheral blood, and EBV strain characterization.

Oropharyngeal EBV excretion was found to persist after the cytoreductive therapy prior to BMT, whereas EBV‐carrying cells in the blood were detected only after 5 weeks following BMT. During the first month post‐BMT, 2 different EBV strains could be isolated from sequential mouthwashes of 3 patients. The initial strains were found to persist up to 7, 21, and 29 days post‐BMT, whereas the subsequent strains appeared at 21, 42, and 34 days post‐BMT, respectively. Thus, the original EBV strain may persist only for a limited time after BMT, and the oropharyngeal epithelium may be reinfected by a new EBV strain from the blood within 3 weeks.

With respect to the coexistence of multiple EBV strains, 2 patterns were evident. From the day 62 mouthwash of 1 patient, 1 Type A and 1 Type B strain were isolated. From the day 180 mouthwash of a second patient, a dominant Type A strain was recovered, together with 6 “variant” strains that differed from each other by only a single EBNA protein (EBNA 1). This pattern may be explained by viral recombinations during replication, which may form the basis for the vast polymorphism of EBV observed in unrelated individuals. © 1992 Wiley‐Liss, Inc.
Infections of the cervix uteri with human papillomavirus and Chlamydia trachomatis
Authors: Claas Eric C. J. , Melchers Willem J. G. , Niesters Hubert G. M. , van Muyden Ruud , Stolz Ernst , Quint Wim G. V.
Published: March 2007

Apart from infection with human papillomavirus (HPV), other microorganisms may be involved in the development of cervical neoplasia. To study concomitant infections with HPV and Chlamydia trachomatis, cervical specimens from 4 groups of women were examined for the presence of these microorganisms by the polymerase chain reaction. The first group consisted of 143 consecutive samples from women with no cytological abnormalities who participated in a triennial screening program to prevent cervical cancer. In this group 2 samples were found positive for HPV and 2 additional samples were found positive for C. trachomatis. In the second group of 46 cytologically abnormal smears, HPV DNA was detected in 71.7% of the samples and C. trachomatis in 4.3%. In a third group of 94 histological abnormal biopsies, the HPV prevalence ranged from 15% in mild dysplastic lesions up to 92% in invasive cervical carcinomas. Only 2 biopsies of this group (2.1%) were found positive for C. trachornatis. Finally, a group of cervical scrapes was obtained from women attending a clinic for sexually transmitted diseases. In 52 samples positive for C. trachornatis and 60 samples negative for C. trachomatis, no significant (P = 0.57) difference in the frequency of HPV infections was found (11.5% and 8.3%, respectively). The data show that in these study groups HPV and C. trachomatis are independently occurring agents. © 1992 Wiley‐Liss, Inc.
Detection of enteroviruses in endomyocardial biopsy by molecular approach
Authors: Petitjean J. , Kopecka H. , Freymuth F. , Langlard J. M. , Scanu P. , Galateau F. , Bouhour J. B. , Ferriere M. , Charbonneau P. , Komajda M.
Published: March 2007

Enteroviruses are considered to be the most common agents implicated in myocarditis and cardiomyopathy. Recent studies have suggested persistent enterovirus infection in chronic disease showing the presence of enteroviral RNA in the myocardium. We used gene amplification by PCR which can demonstrate directly the presence of enteroviral sequences in endomyocardial biopsies. The primers were chosen in the 5′ non‐coding region of the genome representing highly conserved sequences among enteroviruses and therefore allowed the amplification of the majority of enteroviruses. The hybridization of the amplified products was effected with specific general riboprobe derived from 5′ non‐coding sequences internal of the amplified fragments. The results include 105 patients distributed in 6 groups: 45 idiopathic dilated cardiomyopathies with 66.7%, 17 alcoholic cardiomyopathies with 52.9%, 10 myocarditis with 30%, 5 multifactorial cardiomyopathies with 40%, 5 patients with immunosuppressive therapy with 100%, and 23 control group without viral etiology with 39.1% positive samples. The study suggested a positive link between viral infection and cardiomyopathies, but did not allow a direct relation between enterovirus infection and idiopathic dilated cardiomyopathy to be established.
Prevalence of hepatitis C virus antibodies and hepatitis C virus‐RNA in an urban population
Authors: Rapicetta M. , Attili A. F. , Mele A. , de Santis A. , Chionne P. , Cristiano K. , Spada E. , Giuliani E. , Carli L. , Goffredo F. , Capocaccia L.
Published: March 2007

Several studies had been carried out on anti‐hepatitis C virus (HCV) prevalence in populations with blood exposure risks and in blood donors. New tests are now available which allow the in‐ vestigation to extend to other parameters such as antibody type and HCV‐RNA.

In this study the prevalence of anti‐HCV c100‐3 and the associated epidemiological, clinical, and virological markers were evaluated in subjects from an urban population located in central Italy. In positive cases the time persistence of HCV‐ RNA and anti‐HCV antibody pattern was studied. For this purpose, sera from 1,484 randomly sampled individuals, aged 30–69 years, collected in 1985 and stored at −80°C were retrospectively tested. The prevalence was 0.87% (i.e., 13 anti‐HCV c100‐3 positive cases). A significant association was observed with raised alanine transaminase (ALT) levels (P < 0.001). Paired serum samples from 11 out of the 13 subjects collected in 1985 and 1991 were tested by nested polymerase chain reaction (PCR) using primers from the 5′ non‐coding region and by 4‐RIBA. Concordant RIBA patterns between 1985 and 1991 were observed in the majority of positive paired sera (7/9) as well as for HCV‐RNA (619). HCV‐RNA was present in sera simultaneously positive to both types of antibody or to anti‐cl00‐3 or anti‐c22 alone. A wide spectrum of viral and antibody patterns in anti‐HCV c100‐3 positive sera was ob‐ served in this urban population and persisted for at least 6 years. © 1992 Witey‐Liss, Inc. © 1992 Wiley‐Liss, Inc.
Conservation of HPV‐16 E6/E7 ORF sequences in a cervical carcinoma
Authors: Cone Richard W. , Minson Anthony C. , Smith Michael R. , McDougall James K.
Published: March 2007

A cervical carcinoma that contained human papillomavirus (HPV)‐l6 homologous DNA was analyzed. Each tumor cell genome contained a single, incomplete copy of HPV‐16 DNA. The E6 and E7 open reading frames (ORFs) were com‐ pletely conserved relative to other published HPV‐I6 sequences. Much of the non‐coding region (NCR) was free of base changes, including complete conservation of several regulatory elements. Multiple mutations were identified in the remaining integrated HPV‐16 DNA, which was composed of parts of the L1 and E1 ORFs. The extraordinary conservation of the E6/E7 DNA sequence, as compared with other regions of the integrated HPV‐16 DNA, supports the role of E6/E7 in tumorigenesis. © 1992 Wiley‐Liss, Inc.
Donated organ as a source of cytomegalovirus in orthotopic liver transplantation
Authors: Sutherland Sheena , Bracken Paul , Wreghitt Tim G. , O'Grady John , Calne Roy Y. , Williams Roger
Published: March 2007

The importance of the donated organ as a source of CMV was assessed in 120 patients following orthotopic liver transplant and the CMV infections that developed in these patients were graded by severity. Forty‐four recipients were CMV antibody negative pre‐transplant. Eighteen of these received organs from CMV antibody positive donors and 15 (83%) developed primary CMV infections, 13 (87%) of which were symptomatic. Twenty‐six received organs from CMV antibody negative donors and only 2 (8%) became CMV positive post transplant (P < 0.001). These data suggest that there would be a considerable advantage in matching CMV antibody negative recipients with negative donors. Forty‐five percent of secondary infections were asymptomatic compared with 12% of primary infections, and only 11% became disseminated compared with 53% of primary infections. The secondary infections that followed transplantation of an organ from a CMV antibody positive donor were more likely to be symptomatic and were more severe than those in patients who received seronegative livers. © 1992 Wiley‐Liss, Inc.
Pre‐ and posttreatment serum antibody responses to HPV 16 E2 and HSV 2 ICP8 proteins in women with cervical carcinoma
Authors: Lehtinen Matti , Leminen Arto , Kuoppala Tapio , Tiikkainen Mirja , Lehtinen Tuula , Lehtovirta Pentti , Punnonen Reijo , Vesterinen Ervo , Paavonen Jorma
Published: March 2007

Serum antibodies to early proteins of human papillomavirus type 16 (HPV 16 E2 protein) and herpes simplex virus type 2 (HSV 2 ICP8) can be measured by ELISA. In the serum of 122 newly diagnosed cervical carcinoma patients and age‐matched controls, enhanced IgA antibody levels to an HPV‐16 E2 protein derived peptide no. 245 indicated a 9.5‐fold (95% confidence limits 2.8–57.2) relative risk of cervical carcinoma. No significant risk was found with a corresponding HPV 6 E2 peptide or HSV 2 ICP8. To evaluate the HPV 16 E2 peptide as a possible tumor marker for cervical carcinoma serial postoperative serum samples were tested from 27 women with cervical carcinoma. Antibody responses t o the HPV 16 E2 peptide depended on the clinical stage. Stage I and II patients showed decreasing posttreatment IgA and/or IgG antipeptide antibody levels. Stage III and IV patients initially showed decreasing antipeptide antibody levels followed by increasing levels. These patients also showed increasing IgG antibody levels to the HSV 2 ICP8. However, increasing antibody levels to the HPV 16 E2 peptide indicated significantly (P < 0.05) worse 2‐year disease free survival (recurring disease) than did stable or decreasing antibody levels. The results suggest that serum antipeptide antibodies to the HPV 16 E2 peptide no. 245 can be used for the monitoring of cervical carcinoma. © 1992 Wiley‐Liss, Inc.
Sensitivity of an anti‐HCV core peptide ELISA
Authors: Bresters D. , Reesink H. W. , Cuypers H. T. M. , Jansen P. L. M. , Mauser‐Bunschoten E. P. , van der Poel C. L. , Lelie P. N.
Published: March 2007

A newly developed antibody assay based on a synthetic peptide of 15 amino acids derived from the core region of the hepatitis C virus (HCV) genome was evaluated in serum and plasma panels of (A) 225 haemophiliacs and (B) 44 patients with chronic non‐A, non‐B (NANB) hepatitis, and in (C) sequential serum samples of 9 patients with transfusion transmitted HCV infection. The new anti‐core peptide ELISA was compared with the anti‐C100 ELISA. For confirmation of HCV infection, samples were tested in a 4‐antigen recombinant immunoblot assay (4‐RIBA) and samples of panels B and C were also assayed in cDNA‐polymerase chain reaction (PCR). In two panels with a high prevalence of HCV infection (88.4 and 70.5% in haemophilia and NANB hepatitis patients, respectively), the sensitivity of the anti‐core peptide ELISA did not differ significantly from the sensitivity of the anti‐C100 ELISA. The sensitivity of the new assay as compared with the anti‐C100 assay was found to be 0.84 [95% confidence interval (CI): 0.78–0.89] versus 0.92 (95% CI: 0.87–0.95) in haemophilia patients and 0.71 (95% CI: 0.52–0.86) versus 0.84 (95% CI: 0.66–0.95) in NANB hepatitis patients. In sequential serum samples of patients with transfusion‐transmitted HCV infection antibodies to the core peptide (in 6/9 patients) appeared later than antibodies to C100 (in 7/9 patients): 168 (range: 70–322) and 143 (range: 59–365) days after transfusion, respectively. We conclude that although the experimental anti‐HCV assay is based on a single linear epitope from the HCV core region the assay is almost as sensitive for diagnosing HCV infection as the first‐generation anti‐C100 assay. © 1992 Wiley‐Liss, Inc.
Hepatitis C viraemia rebound after “successful” interferon therapy in patients with chronic non‐A, non‐B hepatitis
Authors: Garson J. A. , Brillanti S. , Ring C. , Perini P. , Miglioli M. , Barbara L.
Published: March 2007

A quantitative polymerase chain reaction (PCR) assay for hepatitis C viral RNA (HCV‐RNA) was used to monitor viraemia levels in six patients at multiple time points before, during, and after interferon therapy for chronic non‐A, non‐B hepatitis (NANBH). Prior to therapy, serum HCV‐RNA was detected in all patients at approximately 104‐105 HCV genomes/ml. HCV viraemia became undetectable within 1 month of commencing interferon in three of the five patients whose alanine aminotransferase (ALT) levels decreased to normal on therapy. In the remaining two responder patients, viraemia levels declined more slowly, becoming undetectable after a period of several months. Recurrence of viraemia during therapy was observed in two cases. The one patient whose serum ALT levels remained elevated throughout therapy showed no decline in viraemia. On stopping interferon after a 6 months course, HCV genome titres climbed rapidly in all patients, reaching higher levels than had been observed prior to therapy. Biochemical relapse occurred within 7 months of ending interferon treatment in all but one of the patients who demonstrated this viraemia “rebound” phenomenon. © 1992 Wiley‐Liss, Inc.
Treatment of chronic hepatitis C by continuous subcutaneous infusion of interferon‐alpha
Authors: Carreño Vicente , Tapia Luis , Ryff Jean‐Charles , Quiroga Juan Antonio , Castillo Inmaculada
Published: March 2007

The effectiveness of a daily continuous infusion of interferon‐alpha was evaluated in 12 patients (10 males, 2 females; mean age of 33 years, range 19–62) with biopsy‐proven chronic active hepatitis C. Nine million units (MU) of recombinant interferon‐alpha 2A (rIFN‐α 2A) were administered by continuous subcutaneous infusion with a portable syringe pump, Graseby model MS 16A, for 24 h over 28 days. A significant decrease (P < 0.01) in median serum alanine aminotransferase (ALT) levels was observed after the first week of treatment (96 IU/L, range 58–263) with respect to the pre‐treatment values (188 IU/L, range 119–670). ALT became normal in four patients only by the fourth week. When IFN was interrupted, an increase in ALT was observed in all patients (1.5 to 5 times the pre‐treatment values). The maximum decrease in ALT coincided with a significant increase in serum levels of the enzyme 2′,5′‐oligoadenylate (2‐5A) synthetase (two to fourteen times the pretreatment values) and these parameters were inverse‐correlated (r = −0.598, P < 0.05). 2‐5A synthetase levels returned to pre‐treatment values after discontinuing IFN administration. Hepatitis C virus (HCV) RNA (as detected by the polymerase chain reaction using oligonucleotide primers of the NS5 region) was positive in all cases, remaining so during the treatment period. IgM antibody to HCV (as tested by ELISA) was present in 10112 cases at baseline without changes throughout the study. No irreversible side effects were noted during therapy, which needed to modify the schedule. We conclude that the continuous infusion of 9 MU rIFN‐α 2A/24 h for 1 month is well tolerated by patients with chronic hepatitis C and normalizes ALT levels; although if HCV‐RNA remains detectable at the end of treatment, a rebound in ALT will take place. © 1992 Wiley‐Liss, Inc.
Endonuclease analyses of DNA of human herpesvirus‐6 isolated from blood before and after bone marrow transplantation
Authors: Yoshikawa Tetsushi , Nakashima Toshihiko , Asano Yoshizo , Suga Sadao , Yazaki Takehiko , Kojima Seiji , Mukai Tetsu , Yamanishi Koichi
Published: March 2007

Three strains of human herpesvirus‐6 (HHV‐6) were isolated from peripheral blood mononuclear cells of a leukemic child with the antibody of HHV‐6 before and after bone marrow transplantation (BMT); two strains were obtained before BMT and one after BMT. The DNA extracted from the three isolates was analyzed by six different restriction endonucleases. The cleavage profiles of two strains obtained before BMT were different, but the third strain isolated after BMT was identical with one of the two, which suggest reactivation of HHV‐6 from the recipient's own body after BMT and possible mutation or superinfection of the virus in an immunocompromized patient. © 1992 Wiley‐Liss, Inc.
Peripheral blood leukocytes of patients with argentine hemorrhagic fever as effectors of antibody‐dependent cell‐cytotoxicity
Authors: Ambrosio Ana M. , Gamboa Graciela S. , Feuillade María R. , Briggiler Ana M. , Rimoldi M. T.
Published: March 2007

Peripheral blood mononuclear cells (PBMC) and polymorphonuclear leukocytes (PMNL) of patients with Argentine hemorrhagic fever (AHF) were tested as effectors (E) of antibody‐dependent cell cytotoxicity (ADCC). 51Cr labeled chicken red blood cells (CRBC) coated with anti‐CRBC or normal rabbit serum were used as targets (T).

Three ADCC assays were performed with both effectors from patients: on admission (I), 4 days after the transfusion of immune plasma (II), and 30 days after the clinical onset (III). The ADCC values obtained displayed high variation between individuals. From the linear portions in the curves representing specific 51Cr release vs. E:T ratio plots, extrapolations were made t o determine lytic units (LU), defined here as effector concentrations required t o lyse 50% of the targets. The results were expressed as LU in 106 effector cells.

The killing activity ranges of patients' PMNL (I = 1.04 ± 0.34; II = 2.22 ± 0.66; and III = 2.08 ± 1.18) were not significantly different from that of 21 normal controls (1.19 ± 0.361), except for range II (P < 0.01 1).

ADCC activity ranges of patients' PBMC (I = 3.40 ± 1.06; II = 3.16 ± 1.60; and III = 1.93 ± 0.42) were not significantly different from that determined in 12 healthy subjects (1.86 ± 0.40).

These results demonstrate that patients' PBMC and PMNL can perform ADCC with efficiency comparable to normal effector cells, during the acute period of AHF, and in early convalescence. Consequently, ADCC can be a relevant mechanism in the clearance of Junin virus‐infected cells.
Phytohemagglutinin and concanavalin A activate hepatitis B virus in peripheral blood mononuclear cells of patients with chronic hepatitis B virus infection
Authors: Bouffard Pascal , Lamelin Jean‐Pierre , Zoulim Fabien , Lepot Daniel , Trepo Christian
Published: March 2007

Peripheral blood mononuclear cells (PBMC) from 25 patients with chronic hepatitis B were tested for the presence of free monomeric hepatitis B virus (HBV) DNA migrating as a single 3.2 Kb band by Southern blot analysis. The PBMC were cultured for 7 days in the presence of phytohemagglutinin (PHA) or concanavalin A (ConA) both of which yielded a proliferative response. By contrast, both bacterial lipopolysaccharide (LPS) and interleukin 2 (IL2) failed to do so. Dot blot assays were used to monitor HBV DNA level increase within PBMC. Following mitogen exposure HBV DNA levels increased above pre‐stimulation levels in 19/25 PHA cultures, 6/15 ConA cultures, 1/15 LPS cultures, and 1/15 IL2 cultures.

In 15 patients, Southern blot analysis was carried out before and after PHA exposure. In 13/15 cases, a single 3.2 Kb band was observed in un‐stimulated cultures as well as in PHA cultures even though PHA induced a HBV DNA increase. One case exhibited bands migrating faster than the 3.2 Kb signal, compatible with replicating intermediates and one case provided evidence of viral concatemers within PBMC after PHA stimulation. No HBV DNA was detected in the culture supernatants. The increase of HBV DNA level in PBMC induced by mitogen was strongly associated with an increase in HBV DNA expression (HBV RNA and HBs antigen). These studies indicate that HBV DNA present in human PBMC does represent a potential reservoir for infection with endogenous reactivation following PBMC activation. © 1992 Wiley‐Liss, Inc.
Quantitation of hepatitis C virus RNA by competitive polymerase chain reaction
Authors: Kaneko Shuichi , Murakami Seishi , Unoura Masashi , Kobayashi Kenichi
Published: March 2007

Since the amount of hepatitis C virus (HCV) RNA might be correlated with the degree of severity of hepatitis and response to treatment, quantitation of HCV RNA in serum was established using competitive polymerase chain reaction (PCR). Known amounts of a plasmid containing HCV‐cDNA were co‐amplified with a standard dilution series of a competitive template which shared the primers' sequences but differed from the wild type cDNA in having a deletion. Accurate quantitation was obtained by comparing the amount of both products. Quantitation of serum HCV RNA was carried out in two patients' serum samples which were also used to infect chimpanzees. The concentration of HCV RNA in these two sera was calculated to be 1 pg/ml (non‐infectious at 10−3dilution) and 1–10 pg/ml (infectious at 10−5dilution). The procedure was subsequently used to analyze serial changes in serum HCV RNA in three patients who were treated with a‐interferon. During treatment, the levels of ala‐nine aminotransferase showed a significant decrease in all patients and the amount of HCV RNA fell from 1 fg/ml, 1 pg/ml, and >10 pg/ml to 0.1 fg/ml, 100 fg/ml, and 1 pg/ml, respectively. The decrease in the amount of HCV RNA after treatment was related to the initial amount of serum HCV RNA. These results suggest that quantitation of HCV RNA may be useful not only for understanding the course of HCV infection but also for evaluating treatment for HCV infection. © 1992 Wiley‐Liss, Inc.
Detection of human herpesvirus‐6 DNA by polymerase chain reaction in serum or plasma
Authors: Huang Li‐Min , Kuo Pei‐Fen , Lee Chin‐Yun , Chen Jen‐Yang , Liu Mei‐Ying , Yang Czau‐Siung
Published: March 2007

Human herpesvirus‐6 (HHV‐6) is a newly identified human pathogen. Currently clinicians rely mainly on blood lymphocyte culture and serological tests to diagnose HHV‐6 infection. The polymerase chain reaction (PCR) was carried out on the plasma or sera of patients to determine the value of PCR in the diagnosis of HHV‐6 infection. A total of 30 patients entered the study; 10 were experiencing acute HHV‐6 infections and 20 were healthy and served as controls. HHV‐6 DNA was detected by PCR in the serum or plasma of the 10 cases with acute HHV‐6 infections. All 20 controls had no HHV‐6 DNA in their sera. The time for serum to become PCR‐positive coincided with the appearance of IgG HHV‐6 antibody. The relatively late presence of HHV‐6 DNA in serum might result from late lysis of infected cells by immune responses. It is concluded that detection of HHV‐6 DNA by PCR in the serum is a valuable tool for the diagnosis of acute and/or active viral infection. © 1992 Wiley‐Liss, Inc.
Persistence of hepatitis B virus DNA after reduction of viral replication in serum and liver
Authors: Molina Josefina , Bartolomé Javier , Moraleda Gloria , Ruiz‐Moreno Mercedes , Rua Maria José , Moreno Alberto , Carreno Vicente
Published: March 2007

Liver and serum samples from 67 children with hepatitis B chronic infection, whether or not treated with recombinant interferon, were analyzed for the presence of hepatitis B virus DNA. After follow‐up, 44/67 (66%) still had serum and liver viral DNA; 23/67 (34%) were negative for serum hepatitis B virus DNA. Of the 23 children in the latter group, liver biopsy was available in 21 and viral DNA was not detected by Southernblot in 20. In the remaining patient, viral DNA was in an episomal nonreplicative form. Polymerase chain reaction was performed in the 21 serum samples negative for viral DNA by conventional techniques and in the 21 liver samples (20 negative for hepatitis B virus DNA and 1 with episomal nonreplicative form). All liver samples resulted i n a positive reaction t o viral DNA by this technique. Serum viral DNA by polymerase chain reaction was detected in 15/21 (71%) of these patients. The mean of alanine aminotransferase values was similar in patients with or without hepatitis B virus DNA in serum by polymerase chain reaction. In summary, in the majority of the patients who respond to the therapy, there is a persistence of viral replication detected by polymerase chain reaction. This fact explains the persistence of serum HBsAg in these patients. However, more studies are necessary to determine the meaning of the presence of hepatitis B virus DNA that is only detectable by polymerase chain reaction. © 1992 Wiley‐Liss, Inc.
Genomic variation among herpes simplex virus type 1 strains: Virus DNA analysis of isolates from saudi patients
Authors: Al‐Ahdal Mohammed N. , Kessie George , Taha Mohamed A. , Al‐Shammary Fahad J. , Ettayebi Mohamed
Published: March 2007

Fifty‐two clinical isolates of herpes simplex virus type 1 (HSV‐1) from Saudi Arabian patients were analysed by restriction endonuclease digestion of the virus DNA using the enzymes Hindlll and BarnHI, followed by hybridization with 32P labelled DNA of laboratory strain F. Of the isolates, 17 were resolved into four distinct cleavage patterns with HindIII restriction enzyme. The remaining 35 strains had the same cleavage pattern as the standard HSV‐1‐F. Further investigation of the 52 isolates with BarnHI, which is a multicut enzyme and therefore capable of higher resolution, differentiated 47 of the 52 isolates and were assigned into nine cleavage groups. Comparing our findings with similar studies reported elsewhere suggest geographic clustering of HSV‐1 strains. Fragments giving rise to the observed DNA polymorphism were mapped to the unique region of the long and short segments of the genome. © 1992 Wiley‐Liss, Inc.
Toga virus‐like particles in acute liver failure attributed to sporadic non‐A, non‐B hepatitis and recurrence after liver transplantation
Authors: Fagan Elizabeth A. , Ellis David S. , Tovey George M. , Lloyd Graham , Smith Heather M. , Portmann Bernard , Tan Kai‐Chah , Zuckerman Arie J. , Williams Roger
Published: March 2007

Toga virus‐like particles (typically 60‐70 nm: enveloped with small surface spikes) were detected in the native hepatectomy specimens in 7 of 18 patients grafted for acute liver failure attributed to sporadic non‐A, non‐B hepatitis and in 2 patients grafted for fulminant hepatitis attributed to anti‐epileptic drug hepatotoxicity. These particles were not detected in the hepatectomies from 12 other patients grafted for other causes of acute liver failure, 12 for various chronic liver diseases, and 2 histologically normal livers.

Acute hepatic failure, characterized histologically by severe haemorrhagic necrosis, developed 7 days after grafting in 5 patients, all in the non‐A, non‐B group with toga virus‐like particles in native liver. Similar virus‐like particles were detected in all grafts and were in greater abundance than in the native livers. The agent may be novel because pre‐ and post‐grafting sera were negative for antibodies against representative panels of arboviruses and in first and second generation antibody tests for hepatitis C virus. © 1992 Wiley‐Liss, Inc.
Current seroepidemiology of hepatitis D virus infection among hepatitis B surface antigen carriers of general and high‐risk populations in Taiwan
Authors: Chen Chien‐Jen , Tseng Su‐Feng , Lu Chih‐Feng , Lin Hwa‐Chen , You San‐Lin , Chen Chang‐Sheng , Hwang Shih‐Jen , Hsieh Su‐Fen , Hsu Shu‐Tao
Published: March 2007

In order to assess the current seroepidemiology of hepatitis D virus (HDV) infection in Taiwan where hepatitis B virus (HBV) is hyperendemic, a total of 756 voluntary blood donors, 641 prostitutes, 1,014 patients with sexually transmitted diseases (STDs), and 628 drug abusers were studied. Radioimmunoassays were used for testing HBV infection markers and antibody against HDV (anti‐HDV) among HBsAg carriers. The anti‐HDV prevalence among HBsAg carriers was significantly higher in STD patients (9.6%), prostitutes (33.1%), and drug abusers (68.1%) than in blood donors from the general population (2.2%). The prevalence gradually increased with age in blood donors and STD patients, but reached a plateau at a young age in prostitutes and drug abusers. Males had a higher prevalence than females in blood donors (2.7% vs. 0), STD patients (8.2% vs. 7.5%), and drug abusers (69.0% vs. 57.1%), but the difference was not statistically significant. STD patients with syphilis had a higher prevalence (19.5%) than those affected with non‐ulcerating STDs (5.3%). While unlicensed prostitutes had a lower prevalence (1 3.6%) than licensed prostitutes (44.9%), intravenous drug abusers had a higher prevalence (73.1%) than non‐intravenous drug abusers (34.6%). There was a twofold increase in anti‐HDV prevalence from 1986 to 1989 among prostitutes, but the prevalence remained unchanged in the general population and drug abusers. HDV infection remains limited to the high‐risk groups and spread mainly by promiscuity and needle sharing in Taiwan. © 1992 Wiley‐Liss, Inc.
Passive immunizations of suckling mice and infants with bovine colostrum containing antibodies to human rotavirus
Authors: Ebina Takusaburo , Ohta Minoru , Kanamaru Yoshihiro , Yamamoto‐Osumi Yoshiko , Baba Koichi
Published: March 2007

After immunizing 8‐month pregnant Holstein cows with the human rotavirus MO strain, cow colostrum containing neutralizing antibody to four different serotypes of human rotavirus, designated Rota colostrum, was obtained. Oral inoculation of human rotavirus MO strain into 5‐day‐old BALB/c mice causes gastroenteritis characterized by diarrhea. Using this small animal model, passive protection of suckling mice against human rotavirus infection was achieved with the use of Rota colostrum. Rota colostrums completely protected against rotavirus infection, but purified IgG and IgA obtained from Rota colostrum were unable to protect against infection. After grouping randomly 20 infants from a baby care center, 10 infants received 20 ml of Rota colostrum per day for 2 weeks and 10 control infants did not. Rotavirus‐associated diarrhea developed in 7 of 10 infants in the control group. None of the three infants in the every day recipient group of Rota colostrum had such symptoms, and one of three infants in the every other day recipient group developed rotavirus‐induced diarrhea. All four infants who received Rota colostrum after symptoms appeared developed diarrhea. Oral administration of Rota colostrum seems to be an effective and safe means of preventing diarrhea caused by human rotavirus infection. © 1992 Wiley‐Liss, Inc.
Coxsackievirus B4 infection alters thymic, splenic, and peripheral lymphocyte repertoire preceding onset of hyperglycemia in mice
Authors: Chatterjee Nando K. , Hou Jun , Dockstader Pamela , Charbonneau Tina
Published: March 2007

Diabetogenic Coxsackievirus B4 infection may trigger autoimmune islet loss in diabetes‐susceptible mice, resulting in hyperglycemia in nearly 90% of the animals at 6–8 weeks postinfection (p.i.). To ascertain whether changes in lymphocyte repertoire following infection could predispose these animals to diabetes, alterations in their thymic, splenic, and peripheral lymphocytes were analyzed. Additionally, lymphocyte changes were correlated with the virus load in these tissues and with lymphocyte migration to the inflammatory pancreas. Splenic B lymphocytes more than doubled at 72 hr p.i. and then continuously decreased by 16% of the noninfected controls at 8 weeks p.i. T lymphocytes (CD4+ + CD8+) decreased by about 50% at 72 hr and then increased to the control level by 8 weeks p.i.; CD8 subset continuously decreased by 40% of the control at 8 weeks, resulting in a 67% increase in CD4+/CD8+ ratio. Macrophages and CD5+B subset increased at 72 hr and then dipped by 93% and 84%, respectively, at 8 weeks. In contrast, peripheral B lymphocytes increased by 74% and T lymphocytes decreased by 11% a t 8 weeks p.i. Macrophages increased by twofold at 72 hr and then dipped slightly (6%) at 8 weeks, whereas CD5+B subset increased by 245%. Most prominent thymic T lymphocyte alteration was reflected by about 150% increase in CD4− CD8− cells at 8 weeks p.i. The peak viremia occurred at 72 hr p.i., with highest and lowest virus in the spleen and thymus, respectively. The thymus cleared virus by 3 days, the other tissues by 7 days. lnsulitis and acinar necrosis followed infection; infiltrating lymphocytes were mostly CD4−. Virus‐induced abnormal lymphocyte maturation may contribute to the development of insulitis and hyperglycemia. © 1992 Wiley‐Liss, Inc.
HTLV‐I infection in patients with autoimmune thyroiditis (Hashimoto's thyroiditis)
Authors: Kawai H. , Inui T. , Kashiwagi S. , Tsuchihashi T. , Masuda K. , Kondo A. , Niki S. , Iwasa M. , Saito S.
Published: March 2007

To investigate the possible relationship of HTLV‐I virus infection to autoimmune thyroid disease, we examined, firstly, the frequency of HTLV‐I seropositivity among patients with Hashimoto's thyroiditis and, secondly, the frequency of Hashimoto's thyroiditis in patients with HTLV‐I associated myelopathy/tropical spastic paraparesis (HAM/TSP). Of 144 patients with Hashimoto's thyroiditis in the Tokushima and Kochi Prefectures, Japan, 9 (6.3%) were positive for serum HTLV‐I virus antibody 2 of whom were confirmed histologically to have Hashimoto's thyroiditis. This percentage is significantly higher (P < 0.01) than the estimated prevalence (2.2%) of HTLV‐I carriers among the general population in this region. Of 9 patients with HAM/TSP, 3 (33.3%), including 2 biopsy‐proven cases, had evidence of Hashimoto's thyroiditis. This proportion is apparently much higher than the prevalence (1.7%) of Hashimoto's thyroiditis in the general population. These findings suggest that HTLV‐I virus may be related to the development of Hashimoto's thyroiditis. © 1992 Wiley‐Liss, Inc.
Virus‐like particles with reverse transcriptase activity associated with kawasaki disease
Authors: Lin Ching‐Yuang , Chen I‐Chen , Cheng Tsung‐I , Liu Wu‐Tse , Hwang Betau , Chiang Benjamin N.
Published: March 2007

To investigate the etiologic agent associated with Kawasaki disease (KD), we initially established a cocultivation system using concanavalin A (Con A)‐stimulated lymphoblastoid cells obtained from each retrovirus‐seronegative individual's peripheral blood mononuclear cells (MNCs) cocultivated with each of 1) 40 patients with KD, 2) 10 patients with other viral infection and skin rash, and 3) 10 age‐ and sex‐matched normal controls. Five major findings suggested that virus‐like particles with reverse transcriptase (RT) activity are associated with KD. First, RT activity appeared significantly higher on day 12 after the onset of fever in the KD patients than in those with other viral infections and normal controls (dTMP incorporation: 3,645 ± 248 vs. 434 ± 50 vs. 412 ± 46 cpm, P < 0.0001 1. Second, the RT activity was not endogenous, because the Con A‐stimulated lymphoblastoid cells were obtained from the individuals who were negative for retrovirus. Third, virus‐like particles (80‐100 nm in diameter) by electron microscopy were found in the concentrated pool supernatants of particulate fraction containing RT activity subjected to sucrose density gradient, obtained from KD patients. Fourth, the viral product, a 31.4 kilodalton molecule, was detected by SDS‐PAGE after internal labelling (methionine ‐ S35) and density gradient centrifugation. Fifth, using a “retrovirus universal pol gene region” as a primer and the RT‐PCR method, a retrovirus‐specific band was detected in the cocultivated supernatants obtained from four KD and one AIDS patients but not in patients with rubella or in healthy controls. These findings support the finding that circulating peripheral blood MNCs from KD patients may harbor RT‐associated virus‐like particles containing 31.4 kilodalton molecules. These observations may be of value in identifying the etiological agent and the pathogenesis of Kawasaki disease. © 1992 Wiley‐Liss, Inc.