bluuebunny/crossref_metadata_2025 huggingface dataset:
Temperature enhancement of syncytium formation by HIV and sendai virus
Authors: Kinchington Derek , Barker William , Galpin Sarah , Apostolov Kosta
Published: March 2007

Syncytium formation, the characteristic cytopathic effect (CPE) of the human immunodeficiency virus (HIV) and cell fusion by Sendai virus, is accelerated by increasing the ambient temperature to values at which normal metabolic activity is inhibited. Uninfected C8166, CEM, and H9 cells were absorbed at 4°C onto monolayers of H9 cells chronically infected with HIV and incubated subsequently at either 37°C or 45°C. Similarly chick and human erythrocytes and Hela cells were agglutinated with Sendai virus at 4°C before incubation at temperatures of up to 50°C. With both viruses the rate of cell fusion was directly related to temperature. Since membrane fluidity is dependent on the phase‐transition temperature points of the membrane lipids it is proposed that sufficient membrane fluidity is essential for cell fusion to occur. The implication of these observations on the cytopathology of HIV is discussed.
Characterization of a genetic variant of human hepatitis A virus
Authors: Khanna Bhawna , Spelbring John E. , Innis Bruce L. , Robertson Betty H.
Published: March 2007

Human isolates of hepatitis A (HAV) are a single serotype; however, recent genetic surveys using limited nucleotide sequencing have provided evidence that more than one genotype is responsible for HAV infection in different parts of the world (Jansen et al. [1990]: Proc Natl Acad Sci USA 87:2867‐2871; Robertson et al. [1991] J Infect Dis 163:286‐292). One of these genotypes was originally isolated from Panamanian owl monkeys (strain PA21), but has subsequently been found associated with human cases of HAV from Sweden in 1979 (H‐122) and the United States of America in 1976 (GA76). The nucleic acid sequence of the exposed capsid polypeptide region of GA76 differs from other human HAV sequences by approximately 20%, yet differs by only 2.4% when compared with PI sequence of the PA21 strain. The 20% nucleic acid variability between GA76 and other human HAV results in limited amino acid changes (3%), while a comparison with PA21 revealed only four homologous amino acid substitutions within VP2, VP3, and VP1 polypeptides. HAV infected stool specimens from Nepal and northern India during 1989 and 1990 were found to contain virus whose genetic makeup was related t o the PA21 and GA76 isolates. This genotype of HAV appears to be circulating in some parts of the world where HAV is hyperendemic, and is a potential cause of hepatitis A infection within a susceptible population.
Significance of isolated anti‐HBc seropositivity by ELISA: Implications and the role of radioimmunoassay
Authors: Lai Ching‐Lung , Lau Johnson Y. N. , Yeoh Eng‐Kiong , Chang Wai‐Kwan , Lin Hsiang‐Ju
Published: March 2007

Hepatitis B virus (HBV) surface antigen (HBsAg) and antibody to HBsAg (anti‐HBs) are excellent markers for HBV infection and its immunity. The significance of isolated antibody to HBV core antigen (anti‐HBc) seropositivity is not certain. To elucidate this, sera from 638 Chinese adult subjects, aged 18‐52 years, seronegative for both HBsAg and anti‐HBs, were tested for anti‐HBc. Fifty‐one (8%) were found to have an isolated anti‐HBc seropositivity by ELISA, and all were negative for IgM‐anti‐HBc. The anti‐HBc persisted in all subjects who attended follow‐up for hepatitis B vaccination (n = 48) for a period of 8 months. These 48 subjects received 3 doses of hepatitis B vaccine (HB‐VAX, 10 μg or 20 pg) a t 0, 1, and 6 months: 72.9% developed a primary anti‐HBs response (suggestive of a false‐positive anti‐HBc seropositivity), 4.2% developed an anamnestic or secondary anti‐HBs response, and 22.9% did not develop an anti‐HBs response. Increasing the cutoff point of the ELISA or reconfirmation with radioimmunoassay (RIA) reduced only a minor half of the false positives. This low specificity of anti‐HBc ELISNRIA, together with the high rate of anti‐HBs response to hepatitis B vaccine, indicates that subjects with isolated anti‐HBc seropositivity should be included in vaccination programs.
Serologic identification of hepatitis E virus infections in epidemic and endemic settings
Authors: Favorov M. O. , Fields H. A. , Purdy M. A. , Yashina T. L. , Aleksandrov A. G. , Alter M. J. , Yarasheva D. M. , Bradley D. W. , Margolis H. S.
Published: March 2007

Recombinant chimeric protein C2 containing the N‐terminal region of trpE (37 kilodaltons [kDa]) and the C‐terminal half (46.8 kDa) of the polypeptide encoded by ORF2 of the hepatitis E virus (HEV) genome was used for the construction of a Western blot diagnostic test for IgG and IgM antibodies to the virus (anti‐HEV). (The C2 protein and the trpE protein devoid of C2 activity and used as a control for non‐specific reactions were purified by recovery from sodium dodecyl sulfate‐polyacrylamide gel electrophoresis [SDS‐PAGE] and used for preparation of strips). Specificity of the test was proven with sera obtained from patients with acute hepatitis non‐A, non‐B, non‐C (NANBNC) from outbreaks in different geographic regions of the world. IgG antibodies reactive to the recombinant C2 protein were detected in 93% of patients with acute hepatitis NANBNC and remained detectable in 89‐100% of these patients 1‐24 months after onset of jaundice. IgM antibodies were detected in 73% of patients within 26 days after onset of jaundice, in 50% 1‐4 months after onset, in 6% 6‐7 months after onset, and in no patients by 8 months after onset. When this test was used to identify sporadic hepatitis E cases in different regions of the world, such cases were found almost exclusively in areas where outbreaks of the disease had occurred and rarely in any other regions.
Detection of EBV DNA in post‐nasal space biopsy tissue from asymptomatic EBV‐seropositive individuals
Authors: Lees Janice F. , Goodeve Anne C. , Arrand Jane E. , Ghosh Anna K. , Jones Philip H. , Arrand John R.
Published: March 2007

The association between EBV and nasopharyngeal carcinoma (NPC) has been well documented although the precise role of the virus in the genesis of the tumour is not understood. We undertook this study to examine the prevalence of EBV infection in nasopharyngeal tissue obtained from 33 healthy individuals not considered to be at risk of developing NPC. Using polymerase chain amplification (PCR) and in situ hybridization we have identified EBV DNA in 70% (23/33) of the tissues examined. Our data demonstrate that EBV is present at the site of tumour development in the low‐risk population and by inference that the virus is also present before the onset of disease in the high‐risk group. This survey supports the concept of NPC pathogenesis as a multifactorial process. © 1992 Wiley‐Liss, Inc.
Infection with hepatitis A, B, Delta, And human immunodeficiency viruses in children receiving cycled cancer chemotherapy
Authors: Kumar Archana , Misra P. K. , Rana G. S. , Mehrotra Raj
Published: March 2007

Serological markers of hepatitis A, B, and Delta and human immunodeficiency viruses were studied in 25 children receiving cancer chemo‐therapy. Eighty‐eight percent had pre‐existing HAV immunity which was unaltered by chemo‐therapy. HDV infection was observed in 8% while HIV was conspicuous by its absence. Active HBV infection, observed in 76% of the children, was asymptomatic in the majority and was accompanied by a high incidence of HBe antigenaemia (57.9%) and its persistence. Pre‐existing anti‐HBs failed to prevent HBV infection recurrence, which was, however, transient and self‐limiting. Multi‐ple blood transfusions and repeated parenteral exposures appeared to be the possible sources of HBV acquisition. Transmission to close contacts was also observed. The study suggests that although HBV vaccine might not be protective against HBV infection in patients receiving cancer chemotherapy, it may prevent its persistence and thereby help in reducing chronic liver dis‐ease‐related morbidity and a highly infectious reservoir. Strict HBV screening of blood donors, exclusive use of disposable equipment, and vac‐cination of close contacts of cancer patients is recommended, particularly in HBV endemic third‐world countries. © 1992 Wiley‐Liss, Inc.
Receptor for pre‐Sl(21–47) component of hepatitis B virus on the liver cell: Role in virus cell interaction
Authors: Dash Srikant , Rao Kanury V. S. , Panda Subrat K.
Published: March 2007

Attachment of hepatitis B virus to a hepatoblastoma cell line (HepG2) was examined using a synthetic peptide corresponding to the pre‐S1 (21‐47) region of the envelope protein. Scatchard analysis revealed a single class binding site of Kd 104 ± 27 nM/I and 5.4 ± 1.2 × 105sites per cell. Competition of HBV with pre‐S1 peptides was dose dependent, and demonstrated it as the dominant binding site. In view of the suggested sequence homology between the peptide and IgA, cross‐competition studies were carried out. The results indicate no direct role of IgA receptor in HBV binding. The receptor for the pre‐S1 pep‐ tide was identified as a single major peptide of molecular weight 31 kD using in‐situ ligand receptor crosslinking. © 1992 Wiley‐Liss, Inc.
Seroprevalence of hepatitis C antibody in Peru
Authors: Hyams Kenneth C. , Phillips Irving A. , Moran Alberto Yuen , Tejada Abelardo , Wignall F. Stephen , Escamilla Joel
Published: March 2007

The prevalence in Peru of antibody to hepatitis C virus (anti‐HCV) was determined in a survey of populations living in the northern jungle region and in groups at high risk of parenterally and sexually transmitted diseases. All sera were initially screened for anti‐HCV using commercial first and second generation ELISAs; repeatedly reactive sera were further verified with a second generation immunoblot assay. Serum samples were also tested by ELISA for HBsAg, anti‐HBs, and anti‐HBc. None of 2,111 sera obtained in the survey of jungle residents was positive for anti‐HCV by immunoblot assay. Twelve of 16 HIV‐1antibody positive hemophiliacs, one of 103 HIV‐1 antibody positive homosexuals, and three of 602 HIV‐1 negative registered female prostitutes were positive for anti‐HCV. A high prevalence of total markers of hepatitis B infection was found in all subjects, especially in older subjects and groups at high risk of parenterally and sexually transmitted diseases. The findings of this study indicate that seropositivity for hepatitis C virus antibody is uncommon in Peru except in high risk groups and suggest that the epidemiology of hepatitis C differs substantially from hepatitis B. © 1992 Wiley‐Liss, Inc.
Respiratory syncytial virus detection by dot blot hybridization with a nonradioactive synthetic oligo deoxynucleotide probe
Authors: Hernández Oscar , Fernandez Jorge , Valenzuela Sofia , Sandino Ana Maria , Pizarro Jaqueline , Vasquez Monica , Yudelevich Arturo , Spencer Eugenio
Published: March 2007

A synthetic oligodeoxynucleotide corresponding to a region of the nucleocapside gene (N) of respiratory syncytial virus (RSV), was used as a DNA probe to develop a nonradioactive hybridization assay for the detection of RSV. The probe was labeled by incorporation of biotin‐7‐dATP to the 3′ end by a reaction catalyzed by terminal deoxynucleotydil transferase.

The dot blot hybridization assay was found to be specific for RSV when tested against RSV isolates (subgroups A and B) obtained from cell cultures and isolates of adenovirus, reovirus, rotavirus, and pararotavirus. The assay detected both RSV subgroups (A and B) without significant differences. The dot blot hybridization assay using the nonradioactive probe led to similar results to indirect immunofluorescence (IFI) when tested against a panel of 64 clinical samples from nasopharyngeal secretions of infants with clinical symptoms of respiratory disease. This assay may provide the basis for a rapid, simple, and inexpensive method for routine RSV diagnosis. © 1992 Wiley‐Liss, Inc.
Identification of precursors of structural proteins VP1 and VP2 of hepatitis A virus
Authors: Kusov Yuri Yu. , Kazachkov Yuri A. , Dzagurov Georgy K. , Khozinskaya Galina A. , Balayan Mikhail S. , Gauss‐Müller Verena
Published: March 2007

The morphogenetic pathway of hepatitis A virus (HAV), classified as a member of the enteroviruses within the Picornaviridae, still remains obscure and seems to differ considerably from that of poliovirus, the most studied representative of this genus. In order to elucidate the precursor/product relationship of HAV structural proteins, subviral particles, which represent more than 50% of the viral antigen produced in infected cells, were separated from mature virions and their polypeptide pattern was analyzed by polyacrylamide gel electrophoresis and immunoblotting using monospecific antisera. Whereas mature virions are composed of viral proteins VP1, VP2, and VP3, subviral particles contained VP0 and smaller polypeptides instead of VP2. Comparison of proteins of different strains of HAV showed that VP0 of strain HAS‐15 migrated slower than that of strains MBB or GBM. During the course of the infectious cycle, VP0 accumulated and only small portions were converted to VP2 supporting earlier observations that encapsidation of RNA with concomitant cleavage of VP0 is rate‐limiting, leaving a large amount of viral antigen in premature particles. Similar to VP0, accumulation of VP1 was observed and two immunologically related precursor proteins, p38 and p36, were found during the course of infection. Immunological characterization of p38 using antisera directed to the N‐terminus of VP1 and to synthetic peptides located at the presumptive C‐ and N‐termini of 2A suggests that p38 is VP1Δ2A carrying 45 N‐terminal amino acids of the P2‐region. © 1992 Wiley‐Liss, Inc.
Dideoxyinosine for chronic hepatitis B infection
Authors: Catterall Adrian P. , Moyle Graham J. , Hopes Elaine A. , Harrison Tim J. , Gazzard Brian G. , Murray‐Lyon Iain M.
Published: March 2007

Six patients positive for both human immunodeficiency virus (HIV) and hepatitis B were studied to assess the effect of dideoxyinosine (DDI) on hepatitis B virus (HBV) replication. Two patients died during the follow‐up period and four had at least 8 weeks of therapy. One patient demonstrated HBV DNA suppression and became transiently negative. In the remaining five patients, there was no appreciable change in HBV DNA levels during DDI therapy. DDI was well tolerated in all patients, the only significant side effect being diarrhoea.

It is concluded that DDI has no notable antiviral effect in patients with chronic HBV infection when coinfected with HIV. © 1992 Wiley‐Liss, Inc.
Characterisation by restriction mapping of three subtypes of Molluscum contagiosum virus
Authors: Porter C. D. , Archard L. C.
Published: March 2007

DNA from Molluscum contagiosum virus (MCV) isolates was analysed by restriction endonuclease digestion, identifying three virus subtypes. The structural features of MCV DNA are typical of poxviral DNA. Physical maps of cleavage sites for BamHI, Clal, and Hindlll were constructed for single isolates of each subtype. These differ extensively, indicating the independence of the three subtypes. However, they are closely related, as determined by molecular hybridisation and nucleotide sequence analysis, and their genomes are essentially colinear. There is marked geographical variation in the relative incidence of MCV I and II, whilst MCV Ill is uniformly rare. © 1992 Wiley‐Liss, Inc.
Improved detection of cytomegalovirus viremia in AIDS patients using shell vial and indirect immunoperoxidase methodologies
Authors: Lipson Steven M. , Kaplan Mark H. , Simon Jacob K. , Ciamician Zarui , Tseng Ling‐Fang
Published: March 2007

One hundred twelve peripheral blood specimens were tested for the presence of cytomegalovirus (CMV) by the tube culture indirect immunoperoxidase (TC‐IPA) procedure, the shell vial assay [shell vials were pre‐ and postinoculation treated with medium containing 2 of 10% fetal bovine serum (FBS) or 100 μg% cortisol] (SV‐IFA), and conventional (MRC‐5) tube cultures (TC‐CPE). CMV was detected in 25 (22%) of the 11 2 specimens tested by at least one of these methods. The detection/isolation of CMV among the 25 positive specimens in shell vials maintained with 2% FBS, 100 μg% cortisol + 2% FBS, and 10% FBS was 36,44, and 52%, respectively. Detection/ isolation of the virus from blood by TC‐IPA and TC‐CPE was 52% and 76%, respectively. A significantly greater CMV detection rate occurred using TC‐CPE compared to SV‐IFA treated with medium supplemented with an FBS concentration of 2% (P = .01321, but not medium containing the higher serum supplement or the glucocorticoid (P > .05). Differences in the identification of a CMV viremia were observed by IPA, SV‐IFA, and TC‐CPE methodologies on a patient‐to‐patient basis, denoting the necessity of incorporating each methodology into the CMV screening panel. Demographic analysis of 82 AIDS patients showed a CMV viremia prevalence of 9% (2128) in intravenous drug users, 57% (27/47) in homosexual patients, and 22% (2/9) in heterosexual and transfusion patients, Overnight (24 hr) storage of whole blood at 4 or 24°C, respectively, reduced CMV recovery by 40% and 6570, when tested by TC‐CPE. Improved culture‐based detectioniisolation of CMV in peripheral blood is contingent upon prompt specimen processing, and the utilization of combined SV‐IFA, TC‐IPA, and TC‐CPE methods. © 1992 Wiley‐Liss, Inc.
Monitoring of patients for cytomegalovirus after organ transplantation by centrifugation culture and PCR
Authors: Prösch Susanna , Kimel Viktor , Dawydowa Irina , Krüger Detlev H.
Published: March 2007

A modified centrifugation culture technique and a polymerase chain reaction (PCR) is described for detection of early antigen and IE antigen DNA, respectively, for rapid and sensitive monitoring of active cytomegalovirus (HCMV) infection after organ transplantation.

In a preliminary study, 541 clinical specimens (blood, urine, bronchoalveolar lavage, pharyngeal wash, sputum) from 59 organ recipients were assayed for HCMV antigen by centrifugation culture; 144 samples were tested by PCR simultaneously. Antigenemia detected by centrifugation culture correlated strongly with active HCMV infection and clinical symptoms and proved useful for monitoring the efficacy of anti‐viral therapy. PCR was more sensitive in an earlier phase of infection when centrifugation culture was still negative. The clinical usefulness of both methods is discussed. © 1992 Wiley‐Liss, Inc.
HCMV‐DNA is detected more frequently than infectious virus in blood leucocytes of immunocompromised patients: A direct comparison of culture‐immunofluorescence and PCR for detection of HCMV in clinical specimens
Authors: Ratnamohan V. M. , Mathÿs J. M. , McKenzie A. , Cunningham A. L.
Published: March 2007

In two studies comparing detection of human cytomegalovirus (HCMV) in 118 patients (93 of whom were immunocompromised) by the polymerase chain reaction (PCR) and virus isolation using either early antigen detection by culture‐immunofluorescence or conventional cytopathic effect, DNA‐PCR was found to be the most sensitive, followed by culture‐immunof luorescence, then by cytopathic effect. Urine was inhibitory to the action of Taq polymerase; this was overcome by concentration of HCMV with PEG 6000 prior to gene amplification. Without PEG treatment, HCMV‐DNA in 6 of the 11 specimens positive by culture‐immunofluorescence was not detectable by PCR. In healthy seropositive individuals, HCMV‐DNA was not detected i n leucocytes. However, in immunocompromised patients with AIDS or transplants, and therefore at high risk of HCMV infection or reactivation, blood leucocytes were usually positive for HCMV‐DNA (19/20), some for as long as 20 weeks after initial detection and persisting for long after culture‐immunofluorescence became negative. Neither HCMV‐RNA nor infectious HCMV were detected in the follow‐up blood leucocycte specimens from immunocompromised patients who had detectable HCMV‐DNA in these cells. These data suggest that persistence of HCMV‐DNA in blood leucocytes of immunocom‐promised patients after reactivation or primary infection may be due to persistence of non‐viable virus, residual HCMV genomic DNA, or latent HCMV‐DNA. © 1992 Wiley‐Liss, Inc.
Detection and serotyping of rotaviruses in stool specimens by using reverse transcription and polymerase chain reaction amplification
Authors: Ushijima Hiroshi , Koike Hinako , Mukoyama Atsushi , Hasegawa Ayako , Nishimura Shuichi , Gentsch Jon
Published: March 2007

Direct rotavirus serotyping (VP7, G type) in stool specimens was carried out by reverse transcription and polymerase chain reaction amplification (RT‐PCR) and compared to serotyping by enzyme immunoassay with monoclonal antibodies (EIA‐MAb). The methods used for double‐stranded (ds) RNA extraction, RT‐PCR amplification, and the primers used were modified from previous reports [Gouvea et al.: Journal of Clinical Microbiology 29:519–523, 1990; Gentsch et al.: Journal of Clinical Microbiology, 1992]. For samples that were positive by both methods, the serotypes obtained were identical, however RT‐PCR typing was found to be considerably more sensitive (70.4% samples serotyped) than EIA‐MAb (35.6% of samples serotyped). The overall sensitivities for detection of rotavirus in stool samples by latex agglutination, enzyme immunoassay, electron microscopy, polyacrylamide gel electrophoresis, and RT‐PCR were essentially the same. The results confirm that RT‐PCR typing (genotyping) is extremely valuable for G typing of samples which cannot be typed by EIA‐MAb. We also developed a PCR confirmation technique for serotypes 1, 2, and 4. © 1992 Wiley‐Liss, Inc.
Duration of viremia in human hepatitis a viral infection as determined by polymerase chain reaction
Authors: Yotsuyanagi Hiroshi , Iino Shiro , Koike Kazuhiko , Yasuda Kiyomi , Hino Kunihiko , Kurokawa Kiyoshi
Published: March 2007

Viremia in hepatitis A viral (HAV) infection is said to be limited to pre‐symptomatic period. However, it is not clear how long viremia lasts in human infection due to the lack of a simple and sensitive detection system. In an attempt to find HAV genome in patients′ sera, we used the RTPCR method by setting a pair of primers in the 5′ non‐coding region. While in most cases HAVRNA was detected only before alanine aminotransferase (ALT) reached peak levels with this sensitive system, the viral genome was observed in some patients′ sera even after ALT reached peak levels. Some patients also had HAV viremia after seroconversion to HAV antibody. These results show that viremia in HAV infection lasts longer than has been previously thought, and give a warning of possible secondary blood‐borne infection. © 1993 Wiley‐Liss, Inc.
Evaluation of Phyllanthus amarus and Phyllanthus maderaspatensis as agents for postexposure prophylaxis in neonatal duck hepatitis B virus infection
Authors: Munshi Anupama , Mehrotra Raj , Panda Subrat K.
Published: March 2007

The therapeutic potential of plant extracts of Phyllanthus amarus and Phyllanthus maderaspatensis for postexposure prophylaxis against infection by Hepadnaviruses was studied in ducklings infected by the duck hepatitis B virus (DHBV). Forty‐four Pekin ducklings were inoculated intraperitoneally with DHBV at 24 hr posthatch. They were treated by intraperitoneal injection of Phyllanthus amarus (aqueous extract) (100 mg/kg body weight) or Phyllanthus mad eraspatensis (alcoholic extract) (100 mg/kg body weight) for a period of 4 weeks. Infected ducklings treated with saline served as controls. Weekly serum samples obtained before, during, and after treatment were analysed for the presence of DHBV DNA in serum by dot blot hybridisation using α 32P‐labelled probes. Liver tissue was collected after killing the ducks at various time intervals and was studied for replicative status of the viral DNA and liver histopathology; 17 of 21 ducks were viraemic on completion of treatment with Phyllanthus amarus. At 16 week posttreatment follow‐up four of seven animals remained viraemic. Similar results were obtained with Phyllanthus maderaspatensis. There was no alteration in DHBV replication in the liver. No toxicity was observed with this treatment. These observations suggest that Phyllanthus amarus and Phyllanthus maderaspatensis are not useful as therapeutic agents for postexposure prophylaxis against DHBV infection. © 1993 Wiley‐Liss, Inc.
HCV RNA in patients with chronic hepatitis C treated with interferon‐α
Authors: Liang T. Jake , Rustgi Vinod , Galun Eithan , Blum Hubert E.
Published: March 2007

In order to assess the efficacy of interferon‐α on hepatitis C viral RNA in patients with chronic hepatitis C, we analyzed the levels of HCV RNA in sera and liver tissues of 16 patients pre‐ and posttreatment using a semiquantitative polymerase chain reaction method. Fifteen of 16 patients were positive for anti‐HCV antibodies by four‐antigen recombinant immunoblot assay (4‐RIBA). Only two patients demonstrated normalization of ALT in response to interferon; three patients showed a partial response. Only one patient from the partial responder group displayed a significant reduction of HCV RNA level posttreatment. In the nonresponder group, several patients, although their ALTs remained elevated, demonstrated significant decreases in HCV RNA levels in either serum or liver in response to interferon. Our data suggest that interferon treatment of chronic hepatitis C may not be effective in eradicating HCV infection and a reduction in ALT is infrequently associated with a decrease in HCV RNA level in either serum or liver. Cessation of treatment is frequently associated with the recrudescence of HCV replication and disease. © 1993 Wiley‐Liss, Inc.
Improved detection of rhinoviruses in nasal and throat swabs by seminested RT‐PCR
Authors: Ireland D. C. , Kent J. , Nicholson K. G.
Published: March 2007

A seminested RT‐PCR (nRT‐PCR) was used to detect picornavirus (PV) RNA in cell cultures inoculated with rhinoviruses (HRVs) and enteroviruses (EVs). PCR tests in which a primary "touchdown" PCR was followed by secondary reactions using PV or HRV specific primers were able to differentiate HRVs of 48 serotypes from EVs. PVnRT‐PCR and HRVnRT‐PCR were then used to test nasal and throat swabs from adult subjects with naturally acquired respiratory virus infections. The swabs were also analysed for respiratory viruses by cell culture techniques and the rates of PV identification by the two methods were compared. PVnRT‐PCR was found to be at least five times more sensitive than cell culture for the detection of PVs in these clinical specimens. Paired acute and convalescent serum samples were tested for complement fixing antibodies to adenovirus, influenza A and B, respiratory syncytial virus, parainfluenza viruses 1, 2, and 3, Myco plasma pneumoniae, and Chlamydia psittaci. An enzyme‐linked immunosorbent assay (ELISA) was used to detect rises in antibody level to coronavirus types 229E and OC43. The overall rate of pathogen identification in 159 swabs from adult asthmatics increased from 28% when only cell culture and serology were used to 57% when these methods were supplemented by PVnRT‐PCR. © 1993 Wiley‐Liss, Inc.
Paramyxoviruses and Paget's disease
Authors: Cartwright E. J. , Gordon M. T. , Freemont A. J. , Anderson D. C. , Sharpe P. T.
Published: March 2007

Using the technique of in situ hybridisation, we extended our initial studies of the occurrence of canine distemper and measles virus in Paget's disease. Bone samples from untreated patients and patients treated with 3‐amino‐hydroxypropylidene 1,1 bisphosphonate pamidronate (APD) were examined for canine distemper nucleocapsid (CDV‐N), measles nucleocapsid (MV‐N), respiratory syncytial virus fusion (RSV‐F), and simian virus 5 haemoglutinin‐neuraminidase (SV5‐HN) sequences using 35S‐labelled sense and antisense riboprobes. Only CDV‐N mRNA was detected in osteoclasts, osteoblasts, and mononuclear cells. In untreated Paget's patients, 63.5% were found to have CDV sequences residing in bone cells, and 48.2% of those undergoing APD treatment were still found to have the viral sequences. RSV‐F mRNA was only found in one patient. None of the other paramyxoviruses tested were detected. This study shows that CDV can infect a human host and may be an aetiological agent of Paget's disease of bone. © 1993 Wiley‐Liss, Inc.
Ultra‐rapid, simple, sensitive, and economical silica method for extraction of dengue viral RNA from clinical specimens and mosquitoes by reverse transcriptase‐polymerase chain reaction
Authors: Chungue Eliane , Roche Claudine , Lefevre Marie‐France , Barbazan P. , Chanteau Suzanne
Published: March 2007

A rapid, simple and efficient single‐tube procedure is described for the isolation of dengue virus RNA from small amount of serum (10 μl) followed by a reverse transcriptase‐polymerase chain reaction (RT‐PCR). Recovery of RNA is based on the lysing and nuclease‐inactivating properties of guanidinium thiocyanate in the presence of silica. The silica RT‐PCR can be completed within 5 hours starting from RNA extraction to agarose gel electrophoresis. All of the 63 dengue‐3 culture‐positive sera were RT‐PCR positive (virus titres: <102 to 1110.69). Of 33 culture‐ negative acute sera from serologically confirmed dengue fever patients collected during dengue‐3 epidemic, 4 were RT‐PCR‐positive. RT‐PCR was also positive in 29 of 30 dengue‐1 culture‐ positive sera (virus titres range: <102 to 108.69). Dengue‐1 virus was also detected in fieldcaught Aedes aegypti mosquitoes by silica RT‐PCR. © 1993 Wiley‐Liss, Inc.
Serological responses to human papillomavirus type 16 antigens in women before and after renal transplantation
Authors: Lewensohn‐Fuchs Ilona , Wester Daniele , Bistoletti Peter , Elfgren Kristina , Ohlman Sven , Dillner Joakim , Dalianis Tina
Published: March 2007

Female renal transplant recipients have an increased incidence of human papillomavirus (HPV) associated lesions, such as cervical dysplasia and neoplasia [Schneider et al., 1983]. In this study we tested the serological responses by enzyme‐linked immunoadsorbent assays (ELISA) to 3 different antigenic regions of HPV type 16. Sera from 35 female renal transplant patients collected before and at different times, up to 3 years, after transplantation were collected and tested. Before transplantation IgG antibodies against peptide 49, corresponding to the HPV L2 region, were found in 21/35 of the patients′ sera. Of the L2 positive sera, 16 also demonstrated activity with the HPV L1 region derived peptide 31. All sera that were active against peptide 31 (L1) were also reactive with peptide 49 (L2). After renal transplantation, the antibody levels against these 2 peptides (peptides 49 and 31) dropped significantly (OD ≥ 0.2) in all previously positive sera and remained so throughout the study, which lasted up to 3 years. The proportion of patients with IgA activity against the E2 region (peptide 245), which is common among patients with cervical neoplasia, increased from 9/35 before transplantation to 18/35 after transplantation. In parallel, we monitored 25 of these patients′ sera before and after transplantation for antibody activity against measles, adenoviruses, and cytomegaloviruses (CMV). The majority of these sera—17/25 (68%) and 18/25 (72%), respectively—had no titer changes against measles and adenoviruses. Furthermore, the changes in antibody titers observed with CMV in these patients were not correlated with the fate of the antibodies against the HPV peptides. The results suggest that the interplay between persistent HPV, the host, and the immune system is altered upon therapeutic immunosuppression and that characteristic HPV serological profiles are seen. © 1993 Wiley‐Liss, Inc.
Pathogenesis of the B variant of encephalomyocarditis virus
Authors: Shafi Raheel , Cerutis Delie R. , Giron David J.
Published: March 2007

Variants of encephalomyocarditis virus (EMCV) are immunologically indistinguishable by hyperimmune serum, but, with the exception of EMCV‐B, each produces a different disease syndrome and infects the central nervous system in mice infected via the intraperitoneal route of inoculation. The B variant is benign in that it does not produce any overt signs of infection at doses as high as 106 pfu per animal. The present study was carried out to determine if EMCV‐B was pathogenic when administered via the intracranial route and, if so, to delineate the area(s) of the brain infected. The results show that, when given i.c., EMCV‐B is similar to other variants of EMCV in that it infects and replicates in the brain, causing encephalitis, neuronal necrosis in Ammon's horn of the hippocampus, and clinical signs of infection. The data indicate that receptor sites for EMCV‐B are present on brain cells and suggest that its benign nature when given by the intraperitoneal route reflects an inability to cross the blood‐brain barrier. © 1993 Wiley‐Liss, Inc.
Prevalence, genotypes, and an isolate (HC‐C2) of hepatitis C virus in Chinese patients with liver disease
Authors: Wang Yu , Okamoto Hiroaki , Tsuda Fumio , Nagayama Ryozo , Tao Qi‐Min , Mishiro Shunji
Published: March 2007

China has not been extensively investigated for the prevalence of hepatitis C virus (HCV) infection among people with or without liver disease. We analyzed serum from 2,177 liver disease patients from 7 cities in different areas of China. Of 435 acute hepatitis patients, only 11% were positive for HCV RNA, while hepatitis B surface antigen (HBsAg) was detected in 33%. Of 1,668 patients with chronic liver disease, 14% and 74% were positive for HCV RNA and HBsAg, respectively. Nearly 80% of non‐B chronic liver disease were negative for HCV RNA. The frequency of HCV RNA in chronic liver disease was significantly higher in Hami (32%) and Shenyang (30%) than in other cities (6–12%). The HCV genotype distribution varied by region. Genotype III was detected in 46–70% of HCV infections in Hami, Shenyang, and Lanzhou, while more than 90% of patients from southern cities (Nanjing, Nanning, and Chengdu) had genotype II. No evidence for genotype I or IV infections was found. A fulllength HCV genome sequence (HC‐C2) derived from a Beijing patient with genotype II was closely related to previous isolates from Japanese and Taiwanese patients. These results suggest that HCV prevalence and genotype distribution vary from region to region in China, and that the HCV now predominant in China may have evolved epidemiologically with infections in Japan and Taiwan. The study identified a high frequency of non‐B, non‐C chronic liver disease in China, suggesting possibly a new agent or infections with extreme variants of HCV. © 1993 Wiley‐Liss, Inc.
Diagnosis of enteroviral meningitis with IgG‐EIA using heat‐treated virions and synthetic peptides as antigens
Authors: Samuelson Agneta , Glimåker Martin , Skoog Eva , Cello Jeronimo , Forsgren Marianne
Published: March 2007

Two recently developed enzyme immunosorbent assays (EIA) for the detection of significant titre increases in enteroviral IgG‐antibodies were evaluated as diagnostic tools in 127 etiologically well‐characterized patients with aseptic meningitis. One assay was based on heat‐treated virions (H‐EIA) and one on synthetic peptides (PEIA) as antigens. The sensitivities, with virus isolation as reference method, were 0.67 by H‐EIA and 0.62 by P‐EIA, which were higher than by a routinely used complement fixation test (CFT, 0.51) but somewhat lower than the sensitivities found by two previously presented IgM‐assays, μ‐capture EIA, and solid‐phase reverse immunosorbent test (SPRIST). The specificities of the two IgG‐EIA techniques and CFT were apparently high, whereas the two IgM‐assays showed positive reactions in some non‐enteroviral cases. A relatively rapid increase in enteroviral IgG‐antibodies was apparent using H‐EIA and P‐EIA. The two IgG‐EIA tests contributed with considerable additional etiological information since significant IgG‐rises were obtained in 13 patients by H‐EIA and in 19 by P‐EIA, respectively, out of the 56 individuals in whom enterovirus isolation was negative and a non‐enteroviral diagnosis was not found. Thus, detection of enteroviral IgG‐antibodies by H‐EIA and P‐EIA seems to be a valuable alternative to CFT for the routine diagnosis of enteroviral meningitis. The IgM‐assays, μ‐capture EIA, and SPRIST, may allow a relatively rapid report of an enteroviral infection. However, since both the IgM‐tests are hampered by incomplete specificities, a confirmation of positive results by an IgG‐assay should be carried out. © 1993 Wiley‐Liss, Inc.
Prevalence of hepatitis B, C, and D virus markers in yemeni patients with chronic liver disease
Authors: Guneid Abdulkader M. El , Gunaid Abdalla A. , O'Neill Ann Marie , Zureikat Nael I. , Coleman Jonathan C. , Murray‐Lyon Iain M.
Published: March 2007

A serological survey for hepatitis B, C, and D markers was carried out in the Yemen Republic. Serum samples from 243 pregnant females, 294 male blood donors, and 108 patients with chronic liver disease were examined. Hepatitis B surface antigen (HBsAg) was found in 18.5% healthy individuals and 24.1% patients with chronic liver disease (P = 0.03). Evidence of any marker for hepatitis B virus (HBV) infection was found in 59.8% healthy individuals and 75.9% of patients with chronic liver disease (P = 0.0016). HBeAg was detected in 32.1% of the HBsAg‐positive pregnant females, indicating that vertical transmission probably plays a part in forming the pool of HBV carriers. Vaccination against HBV as part of the extended programme of immunisation (EPI) is recommended. Antibodies to hepatitis D were found in only 2 of 100 HBsAgpositive sera. Antibodies to hepatitis C (anti‐ HCV) were found in 2.1% healthy individuals and 21.5% patients with chronic liver disease (P = 0.0001).

These results indicate that hepatitis B is hyperendemic in the Yemen Republic but that hepatitis D is very uncommon. The prevalence of anti‐HCV is higher than in Europe and similar to neighbouring Arab countries. Infection with both HBV and HCV are important causes of chronic liver disease in the Yemen Republic.© 1993 Wiley‐Liss, Inc.
Detection of hepatitis C virus genome in human serum by multi‐targeted polymerase chain reaction
Authors: Okamoto M. , Baba M. , Kodama E. , Sekine K. , Takagi T. , Kasukawa R. , Shigeta S.
Published: March 2007

A multi‐targeted “hemi‐nested” PCR (M‐PCR) assay in which the primer pairs derived from the 5′ non‐coding (5′NC) and the nonstructural protein 3 (NS3) regions of HCV genome were concurrently used for amplification in order to compare the sensitivity and specificity of polymerase chain reaction (PCR) with different primer pairs in detecting hepatitis C virus (HCV) genome. Sera from patients with virus‐associated liver diseases were examined for the presence of HCV RNA by the M‐PCR method following reverse transcription to cDNA. The amplified products derived from both the 5′NC and the NS3 regions were detected in 28 (70%) of the 40 HCV RNA‐positive samples. However, 12 samples (30%) were devoid of the signal of NS3‐derived product. Sensitivity tests using serial dilutions of HCV RNA revealed that the 5′NC‐derived band was still detectable in the 105‐fold diluted sample by the M‐PCR method, yet the NS3‐derived band could hardly be detected in the 104‐fold diluted sample. Thus, as previously demonstrated by a single‐targeted “nested” PCR assay, the present study using the M‐PCR assay has clearly shown that the 5′NC‐derived primers are more sensitive and specific than the NS3‐derived primers in detecting HCV RNA.
Cytomegalovirus recurrence in seropositive pregnant women attending obstetric clinics
Authors: Shen Chen‐Yang , Chang Shu‐Fen , Chao Maio‐Fen , Yang Show‐Lin , Lin Gin‐Mei , Chang Whi‐Wen , Wu Cheng‐Wen , Yen Ming‐Shyen , Ng Heung‐Tat , Thomas James C. , Huang Eng‐Shang
Published: March 2007

To understand reasons for cytomegalovirus (CMV) recurrence, a cohort of 350 CMV‐seropositive pregnant women attending obstetric clinics in Taiwan was examined for cervical or urinary CMV shedding. Urine specimens were collected from 350 women and cervical secretion specimens were collected from 220 women. We measured the association of various factors with CMV recurrence, which was defined as viral shedding identified by the presence of a CMV‐specific gene sequence amplified by the polymerase chain reaction in seropositive individuals. CMV recurrence status was independently associated with a sexual activity composite variable, which was defined by three sexual activity indicators: age at first sexual intercourse, total years of sexual experience, and average frequency of sexual intercourse prior to pregnancy. Pregnant women with a history of genital tract infection were more likely than women without such history to experience cervical CMV recurrence. Similarly, pregnant women with previous urinary tract infections were more likely to experience urinary CMV recurrence. The findings indicate that multiple exposure to CMV by sexual activity prior to pregnancy is an important determinant of CMV recurrence during pregnancy.
Is there a relationship between malnutrition, inflammation, and post‐vaccinal antibody response to influenza viruses in the elderly?
Authors: Pozzetto Bruno , Odelin Marie‐France , Bienvenu Jacques , Defayolle Michel , Aymard Michèle
Published: March 2007

This study was designed to explore the relationship between malnutrition, inflammation, and the specific antibody response after influenza vaccination in the elderly. Eighty‐two aged subjects, immunized annually against influenza with a trivalent inactivated vaccine, were evaluated for 9 protein markers (albumin, thyroxin‐binding prealbumin, transferrin, immunoglobulins (Ig) G, M, and A, orosomucoid, haptoglobin, and C reactive protein) and for their antibody response to influenza viruses in comparison to 29 younger adults who received the same vaccine and 21 unvaccinated adults. IgM and nutritional markers were significantly reduced in the aged as compared to controls, while the opposite pattern was seen for IgA and inflammatory markers. No difference was observed between the elderly and the controls with regard to the antibody response to influenza virus after vaccination. Reciprocally, influenza immunization had no influence on the levels of the protein variables. These results suggest that the protein status does not play an important role in the antibody response to influenza vaccination in the elderly, a fact which could be related to the slight involvement of cellular immunity in the defense against influenza reinfection.
Infection of human monocyte‐derived macrophages by human immunodeficiency virus mediated by cell‐to‐cell transmission
Authors: Luce Serena St , Arts Eric , Geleziunas Romas , Bour Stéphane , Wainberg Mark A.
Published: March 2007

We have infected ten‐day‐old primary cultures of human monocyte‐derived macrophages (MDM) with HIV‐1 by cocultivation with chronically infected monocytic cell lines. This work has involved the U‐937 monocytoid cell line, chronically infected with the HIV‐IIIB strain of HIV‐1 (U‐937HIV IIIB) as well as a number of cell clones, termed UHC, which were derived from U‐937HIV IIIB by limiting dilution. Cell‐free virus, derived from each of U‐937HIV IIIB cells and the UHC1 clone were noninfectious for MDM, as determined by failure to express viral p24 antigen (Ag). In contrast, viral p24 Ag production was detected in MDM that had been cocultivated with U‐937HIV IIB, and with each of three UHC clones that produced infectious virus. Infection, in each case, was confirmed by polymerase chain reaction detection via the amplification of proviral DNA. In contrast, cocultivation with the UHC15.7 clone, which fails to cleave viral gp160 to its gp120 and gp41 products or the UHC8 clone, which lacks functional reverse transcriptase, did not lead to infection of MDM. Pretreatment of MDM for 2 hr with 1 μM AZT completely prevented infection by culture fluids containing HIVada' a macrophage‐tropic virus, but did not affect infection mediated by cocultivation. These results suggest that cell‐to‐cell transmission of HIV‐1, among monocyte‐derived macrophages, can be mediated by proviral DNA. Moreover, gp120 at the surface of infected cells may play an important role in this process, since cell‐to‐cell HIV transmission could not be demonstrated with the UHC clone that is defective in cleavage of the viral envelope glycoprotein gp160.
Detection of human papillomavirus L1 protein in condylomata acuminata from adults with defects in cell‐mediated immunity
Authors: Brown Darron R. , Bryan Janine T. , Wools Kara , Rodriguez Michael , Tyring Stephen
Published: March 2007

Immunohistochemical assays for human papillomavirus (HPV) L1 protein, using antiserum directed against the L1 major capsid protein of bovine papillomavirus (anti‐BPV serum), were performed on 101 condylomata acuminata biopsy samples from 47 men (40 of whom had intact cell mediated immunity [CMI], and 7 with conditions known to cause CMI defects), and 54 women (48 with normal CMI, and 6 with CMI defects). L1 protein was detected in 28% of all biopsies, including 20.5% of samples from patients with normal CMI and 76.9% of patients with CMI defects (P = .00002). For both males and females, L1 protein was detected significantly more often in samples from patients with CMI defects than those with normal CMI. Immunohistochemical assays of HPV 11‐infected human foreskin implants grown in athymic mice were performed to optimize the conditions of the assay. Three dilutions of anti‐BPV serum or preimmune rabbit serum were used. A 1:500 dilution provided readily interpretable results, while preimmune serum at this dilution did not stain to any significant degree. However, at 1:10 and 1:100 dilution, both the anti‐BPV serum and preimmune serum caused an unacceptable amount of nuclear staining, making results uninterpretable. Defects in CMI may allow active viral particle assembly at an increased rate, as judged by detection of L1 protein in condylomata acuminata from these patients.
Immunohistological detection of human herpes virus 6 in formalin‐fixed, paraffin‐embedded lung tissues
Authors: Pitalia Anil K. , Liu‐Yin John A. , Freemont Anthony J. , Morris David J. , Fitzmaurice Richard J.
Published: March 2007

Human herpes virus type 6 (HHV‐6) infection is widespread in healthy individuals. The only definite disease association is with exanthem subitum in infants though the virus has been linked with a variety of other diseases including interstitial pneumonitis in bone marrow allograft recipients. In order to investigate the role of HHV‐6 in the latter disease we have developed an optimised staining method for the demonstration of specific antigen in routinely processed post‐mortem tissues. Formalin‐fixed, paraffin‐embedded lung tissue from 8 immunocompromised patients who died from interstitial pneumonitis was subjected to immunoperoxidase staining with monoclonal antibodies against HHV‐6, cytomegalovirus (CMV), and adenovirus, using a modified avidin‐biotin complex (ABC) method. Staining for HHV‐6 was obtained in 6 of the 8 patients studied and was present in pneumocytes and macrophages. CMV and adenovirus antigens were identified in 4 and 6 patients, respectively. Whilst the lung tissue of 6 patients contained more than one virus, there was no evidence of cross‐reactivity between the monoclonal antibodies. We demonstrated that accurate localisation of HHV‐6 using monoclonal antibodies is possible in post‐mortem lung tissue and conclude that either HHV‐6 alone or in combination with other viruses may play a role in the development of interstitial pneumonitis following bone marrow transplantation or chemotherapy.