bluuebunny/crossref_metadata_2025 huggingface dataset:
Inhibition of hepatitis B virus DNA polymerase by 3′‐fluorothymidine triphosphate and other modified nucleoside triphosphate analogs
Authors: Meisel H. , Reimer K. , Janta‐Lipinski M. V. , Bärwolff D. , Matthes E.
Published: March 2007

The 3′‐fluoromodified nucleotide analogs 3′‐fluorothymidine triphosphate (FdTTP), 2′,3′‐dideoxy‐3′‐fluoro‐5‐chlorouridine triphosphate (F‐5CldUTP), 2′,3′‐dideoxy‐3′‐fluoro‐5‐ethyluridine triphosphate (F‐5EtdUTP), 2′,3′‐dideoxy‐3′‐fluorouridine triphosphate (FdUTP), and 2′,3′‐dideoxy‐3′‐fluoro‐5‐fluorouridine triphosphate (F‐5FdUTP) as well as 2′,3′‐dideoxythymidine triphosphate (ddTTP), 2′,3′‐didehydro‐2′,3′‐dideoxythymidine triphosphate (ddeTTP), 3′‐chlorothymidine triphosphate (CldTTP), and 3′‐rhodanothymidine triphosphate (SCNdTTP) were tested for their ability to inhibit hepatitis B virus (HBV)‐associated DNA polymerase activity in vitro. The ID50 values of the most potent inhibitors were 0.15 μM for FdTTP, 0.2 μM for ddeTTP, 0.45 μM for ddTTP, and 0.8 μM for F‐5CldUTP. SCNdTTP, CldTTP, and F‐5EtdUTP were less efficient (ID50 = 3–5 μM), and FdUTP and F‐5FdUTP were the least efficient inhibitors (ID50 = 25 μM) of the enzyme activity. Kinetic analysis revealed a competitive type of inhibition for FdTTP and ddeTTP. The Ki values were estimated to be 0.04 μM and 0.08 μM, respectively, compared with a Km value for dTTP of about 0.18 μM.
Anti‐PreS2 antibody assay for evaluating immune responses among recipients of recombinant hepatitis B PreS2+S vaccine
Authors: Hurni William M. , Roehm Robert R. , Miller William J.
Published: March 2007

A competitive radioimmunoassay was developed for measuring hepatitis B virus (HBV) anti‐PreS2 antibody. The assay has been demonstrated to be highly specific for anti‐PreS2 antibody and not subject to interference by other antibodies to HBV‐specific antigens. This assay was used to evaluate anti‐PreS2 antibody responses in a hepatitis B PreS2+S vaccine human clinical trial in healthy adults.
Protracted alanine aminotransferase levels in tamarins infected with hepatitis A virus
Authors: Karayiannis P. , Chitranukroh R. , Fry M. , Petrovic L. M. , Moore D. , Scheuer P. J. , Thomas H. C.
Published: March 2007

Eighteen red‐bellied tamarins (Saguinus labiatus), experimentally infected with hepatitis A virus (HAV), were followed for up to 1.5 years after initial challenge. Half of these animals developed protracted alanine aminotransferase (ALT) abnormalities, which lasted for between 23 and 55 weeks post‐challenge. IgM anti‐HAV was detected intermittently during the early phase of their relapsing hepatitis and never after return of ALT levels to normal. The possibility that these findings may be related to continued HAV replication, coinfection with another agent, or the result of an autoimmune phenomenon triggered by HAV are discussed.
Hepatitis B virus DNA in peripheral blood leukocytes: A brief review
Authors: Harrison Tim J.
Published: March 2007

Despite being essentially hepatotropic, hepatitis B virus has frequently been detected in peripheral blood leukocytes. In most cases, viral DNA has been detected by hybridisation, though, occasionally, evidence of viral RNA and virus‐specific proteins has been reported. Though definitive evidence is lacking, it appears likely that the virus is able to replicate in at least a subfraction of these cells. The possible consequences with respect to the transmission and reactivation of virus infections are discussed.
Hepatitis B virus infection of peripheral blood mononuclear cells is common in acute and chronic hepatitis
Authors: Pasquinelli Claudio , Melegari Margherita , Villa Erica , Scaglioni Pier Paolo , Seidenari Michela , Mongiardo Nicola , De Rienzo Bruno , Manenti Federico
Published: March 2007

Hepatitis B virus (HBV) infection of peripheral blood mononuclear cells (PBMCs) has been observed in all stages of liver disease. Thus far all information about the physical state of HBV in mononuclear blood cells comes from Southern blot analysis and in situ hybridization. In this study we focused our attention on the presence of HBV DNA sequences in PBMCs of 30 patients with acute type B hepatitis and 6 patients with chronic active hepatitis by utilizing both Southern blot analysis and the polymerase chain reaction (PCR). Southern blot analysis showed no HBV DNA sequences in PBMCs of the acute hepatitis patients, although the sensitivity of our method enabled us to detect as little as 1 pg of cloned HBV insert. As far as the chronic hepatitis patients are concerned Southern blot analysis revealed the presence of HBV DNA sequences in 5 out of 6 patients but intermittently at successive follow‐up times. On the other hand we were able to demonstrate the presence of HBV related sequences in 14 out of 30 acute hepatitis patients (5 HBeAg positive, 9 antiHBe positive) and in all 6 chronic hepatitis patients by PCR. Our results indicate that the involvement of PBMCs with HBV during acute HBV infection occurs at a very low level, often below the detection limit of the Southern blot technique.
Cultivation of human immunodeficiency virus from whole blood: Effect of anticoagulant and inoculum size on virus growth
Authors: Bayliss Gary J. , Jesson William J. , Mortimer Philip P. , McLean Kenneth A. , Evans Brian A.
Published: March 2007

Human immunodeficiency virus (HIV) was cultivated directly from whole blood treated with anticoagulant by cocultivation with phytohaemagglutinin‐stimulated cord blood lymphocytes. When heparin was used as the anticoagulant, isolation rates were low (10% to 56%, depending on the patient group); but when EDTA was used, isolation rates were much higher (50% to 100%). Culture of whole blood gave results identical to those of culture of separated peripheral mononuclear cells, and in some cases virus could be isolated from as little as 10 μl of unseparated EDTA anticoagulated blood.
Virus‐like particles in the liver of a patient with fulminant hepatitis and antibody to hepatitis E virus
Authors: Shavrina Asher Ludmila V. , Innis Bruce L. , Shrestha Mrigendra P. , Ticehurst John , Baze Wallace B.
Published: March 2007

In earlier studies, hepatitis E virus (HEV) particles were detected in the stools of patients with enterically transmitted non‐A, non‐B (ENANB) hepatitis, and HEV was etiologically associated with this disease. Such particles have not been observed in the liver, however. We describe the pathological findings in the liver of a young pregnant woman from Nepal who died as a result of fulminant NANB hepatitis. IgM antibody to HEV was detected in the patient's serum by immune electron microscopy, suggesting that she was acutely infected with that virus. On light microscopic examination of the liver we observed cholestatic hepatitis with proliferation of bile ductules and pseudoglandular arrangement of hepatocytes around distended bile canaliculi. Three types of virus‐like particles were detected by electron microscopy. The most frequently observed particles were in cells lining small bile ductules; they measured 32–37 nm and were enclosed by a membrane. Particles of a second type were seen in clusters in the sinusoidal cells; they were uniform in size, without a membrane, and measured about 32 nm in diameter. Particles of a third type (65 nm) were found in epithelial cells of the small bile ductules. Among the particles we detected, the 32 nm particles most closely resembled those of HEV.
HIV excretion patterns and specific antibody responses in body fluids
Authors: O'Shea Siobhan , Cordery Mark , Barrett Winsome Y. , Richman Douglas D. , Bradbeer Caroline , Banatvala Jangu E.
Published: March 2007

HIV excretion patterns and specific antibody responses were evaluated in blood, semen, female genital secretions, saliva, and crevicular fluid. Samples were examined for infectious virus, viral antigens, viral nucleic acid, HIV specific IgG, IgA, anti‐nef, and anti‐p24. Viral load in peripheral blood appeared to increase with disease progression. The proportion of patients with antibody responses specific for nef and p24 was also lower among patients with more advanced disease. Infectious virus and viral antigens were detected infrequently and at lower levels in body fluids than in blood, which may reflect the presence of local antibodies. HIV nucleic acid was detected in some semen and saliva samples in the absence of infectious virus.
Molecular epidemiology of Australian isolates of molluscum contagiosum
Authors: Thompson Carol H. , De Zwart‐Steffe Rita T. , Biggs Ian M.
Published: March 2007

Molluscum contagiosum lesions obtained from 75 Australian patients, 22 (29%) of whom were HIV positive, were examined by restriction endonuclease analysis and Southern blot hybridisation using radiolabelled and digoxigeninlabelled MCV DNA probes. The isolates were classified (MCV 1, 1v, 2, and 2v) on the basis of these results, which were in turn correlated with the clinical features of each lesion. A total of 44 (59%) of the lesions contained MCV 1 or 1v, 24 (32%) contained MCV 2 or 2v, three (4%) contained multiple MCV types, whereas four (5%) of lesions submitted contained no detectable MCV DNA. The ratio of MCV 1/1v to MCV 2/2v was determined to be approximately 1.75:1. There were substantial differences between the distribution of MCV types 1 and 2 among patients of different age groups, but no significant relationship between MCV type and the sex of the patient was found. MCV 2 was more frequently detected in lesions from anogenital areas and in immunosuppressed (HIV‐positive) patients, and MCV 1 was more commonly isolated from skin rather than genital lesions, but neither association was statistically significant. Fragment F (10 kbp) obtained from the genome of MCV 1 was capable of differentiating MCV 1 from MCV 2 when used as a probe in hybridisation experiments with BamHl cut samples and may be useful when small amounts of lesion prevents differentiation by direct visualisation of restriction endonuclease fragments.
Incidence of antibodies to human papillomavirus type 1 in patients with cutaneous and mucosal papillomas
Authors: Viac Jacqueline , Chomel Jean‐Jacques , Chardonnet Yvette , Aymard Michèle
Published: March 2007

A study of antibodies to human papillomavirus (HPV) 1 is reported in a series of sera from 126 patients with papilloma lesions of different clinical types (verruca plantaris, vulgaris, plana, condyloma acuminata, laryngeal papilloma) using a sensitive solid‐phase enzyme‐linked immunosorbent assay (ELISA). Our findings showed a significant prevalence of specific HPV 1 IgG antibodies in patients with plantar warts (65%). This incidence was considerably lower in patients with other papilloma lesions, in which positive sera may reflect a past infection with HPV 1. The ELISA test was evaluated as nearly 100 times more sensitive than the indirect immunofluorescence test, and lgG titers to HPV 1 in patients' sera were usually low. Serum samples taken at different times during the course of HPV infection in a few patients with refractory lesions did not show any significant change in their antibody titer.
Automated polymerase chain reaction for papillomavirus screening of cervicovaginal lavages: Comparison with dot‐blot hybridization in a sexually transmitted diseases clinic population
Authors: Morris Brian J. , Rose Barbara R. , Flanagan Judith L. , McKinnon Kay J. , Loo Cheen Y. , Thompson Carol H. , Flampoulidou Mary , Ford Robert M. , Hunter J. Christopher , Nightingale Brian N. , Cossart Yvonne E.
Published: March 2007

The aim of the present study was to compare the recently developed polymerase chain reaction (PCR) technique with conventional dot‐blot DNA hybridization for human papillomavirus (HPV) detection. Cells were collected by cervicovaginal lavage from a study group of 109 women attending a sexually transmitted diseases clinic. Using a machine that we developed for alternation of temperature cycles, HPV was detected in 51% of patients by PCR. By dot‐blot hybridization, 44% of the patients were positive. Concordance of combined positive and negative results between PCR and dot blot was 69%. The greater sensitivity of PCR may have accounted for 19% of specimens that were PCR positive but dot‐blot negative. Unexpectedly, however, 12% of specimens were dot‐blot positive but negative by PCR, and several specimens were discordant for type of HPV. Both HPV DNA tests agreed with cytology in 41% of women, and in 33% cytology was negative in the face of positive PCR and dot blot. Concordance of cytology with just PCR was 59%, and only with dot blot was 56%. Cervicography agreed with both HPV DNA tests in 41% of patients, with PCR alone in 55%, and with dot blot alone in 58%. Biopsy results did not reveal a strong correlation between histopathological criteria of HPV infection and detection of HPV DNA by either PCR or dot‐blot hybridization. Thus the present study has shown that PCR is a slightly more sensitive indicator of HPV infection than dot‐blot hybridization. Agreement of HPV DNA results with conventional screening tests was not strong, an observation consistent with many comparative studies by others. In conclusion, PCR is slightly more sensitive than DNA hybridization for detection of HPV, it can be used in conjunction with specimen collection by gentle lavage of the cervicovaginal epithelium, and the possibility remains that it may prove suitable as a screening test.
Detection of human herpesvirus‐6 DNA in peripheral blood and saliva
Authors: Jarrett Ruth F. , Clark Duncan A. , Josephs Steven F. , Onions David E.
Published: March 2007

Saliva and peripheral blood samples from 20 healthy adults were examined for the presence of human herpesvirus‐6 (HHV‐6) DNA sequences using the polymerase chain reaction. Eighteen out of 20 whole saliva samples contained detectable HHV‐6 genomes. The majority of peripheral blood samples were also positive; however, the results suggest that only a rare cell in the peripheral blood is infected. Serological studies did not reveal any correlation between HHV‐6 antibody titre and the ability to detect HHV‐6 DNA. The data indicate that HHV‐6 genomes persist in the peripheral blood and oropharynx or salivary glands of most healthy individuals following primary infection.
Hepatitis B pre‐S1 and pre‐S2 proteins: Clinical significance and relation to hepatitis B virus DNA
Authors: van Ditzhuijsen T. J. M. , Kuijpers L. P. C. , Koens M. J. , Rijntjes P. J. M. , van Loon A. M. , Yap S. H.
Published: March 2007

Pre‐S proteins may have an important role in virus assembly and virus entry into the host cell. The presence of pre‐S proteins in serum has also been thought to correlate with active viral replication. To investigate whether pre‐S proteins in serum might have additional diagnostic and/or predictive value for liver sequelae in HBV infection, sera from six different serological groups of patients with HBV markers (total number 363) and different manifestations of liver histology were examined for the presence of pre‐S1 and pre‐S2 proteins using micro‐ELISAs. Pre‐S1 and pre‐S2 proteins were detected significantly more often in HBV‐DNA‐positive than in HBV‐DNA‐negative sera from HBsAg carriers. However, pre‐S1 and pre‐S2 proteins were also found in HBV‐DNA‐negative HBsAg carriers irrespective of serum HBeAg/anti‐HBe or liver histologic findings. These results suggest that the presence of the pre‐S1 and or pre‐S2 proteins in serum either does not seem to reflect the presence of active viral replication and active liver disease or pre‐S proteins are more readily detectable than HBeAg and HB‐DNA as measured by a dot‐blot technique. Furthermore, the presence of pre‐S proteins in serum is strongly correlated with that of HBsAg.
Antigenic relatedness among the Norwalk‐like agents by serum antibody rises
Authors: Madore H. Paul , Treanor J. J. , Buja R. , Dolin R.
Published: March 2007

The Norwalk, Snow Mountain (SMA), and Hawaii agents are etiologically associated with separate outbreaks of acute viral gastroenteritis. Previous cross‐challenge of volunteers, immune electron microscopy, and/or enzyme‐immunoassay analysis suggested that these agents are antigenically distinct. We examined paired sera from human volunteers challenged with these agents for the presence of homologous and heterologous serum antibody titer rises to the agents. Two‐way cross‐reactions occurred between Hawaii agent and SMA. A one‐way cross‐reaction occurred between Norwalk agent and SMA, as volunteers challenged with Norwalk agent had heterologous serum antibody titer rises to SMA, but the reverse did not occur. The Norwalk and Hawaii agents had minimal cross‐reaction, with only one volunteer challenged with Hawaii agent having a heterologous rise to Norwalk agent. These observations indicate varying degrees of antigenic relatedness among these agents.
Diagnosis of cytomegalovirus infection in cyclosporin‐treated renal allograft recipients
Authors: Morris D. J. , Fox A. J. , Klapper P. E.
Published: March 2007

The relative merits of antibody response and virus shedding as markers of cytomegalovirus (CMV) infection among cyclosporin‐treated renal allograft recipients were analysed. CMV‐specific antibody was assayed by lgG‐specific radioimmunosorbent test (RIST) and by complement fixation test (CFT). CMV shedding was assayed by virus isolation and by the rapid test immediate early nuclear antigen detection (IENAD). RIST and CFT detected seroconversion in similar numbers of patients, but the former test was the more sensitive when CMV antibody was sought in pretransplant sera to differentiate primary from recurrent infection. IENAD detected or excluded CMV shedding for more urine specimens than virus isolation (462/515 [90%] vs. 366/515 [71%]), but the reverse applied to saliva specimens (33/57 [58%] vs. 54/57 [95%]). The high specificity of IENAD allowed positive results by IENAD to be accepted when virus isolation failed to provide a result. IENAD was, however, less sensitive than virus isolation even when specimens yielding CMV by IENAD, but no result by virus isolation, were included in the analysis (27/44 [61%] vs. 38/44 [86%]). Assays of both antibody response and virus shedding were required to maximise the diagnosis of recurrent CMV infections, but the former assay detected all primary CMV infections. The diagnostic implications of these results are discussed.
Neopterin and beta‐2 microglobulin levels in asymptomatic HIV infection: The predictive value of combining markers
Authors: Harrison Nigel A. , Skidmore Susan J.
Published: March 2007

The concentrations of neopterin, beta‐2 microglobulin, p24 antigen anti‐p24 antibody, and alpha‐interferon were determined in the sera of 25 asymptomatic anti‐HIV positive patients (CDC stages II and III) over a 50 month period (38 months retrospectively and 12 months prospectively). Evaluation of the data allowed the derivation of a threshold of abnormality for both neopterin and beta‐2 microglobulin. Pre‐AIDS (CDC groups IVa, IVc2, IVe) developed in 6 of the 26 patients within 18 months of the first abnormal value of beta‐2 microglobulin, and within 12 months of the first abnormal value of neopterin. Combining the abnormal results of neopterin, beta‐2 microglobulin, p24 antigen, and alpha‐interferon allows prediction of progression to pre‐AIDS within 18 months for most asymptomatic patients.
Development and evaluation of the TD97 measles virus vaccine
Authors: Suzuki K. , Morita M. , Katoh M. , Kidokoro M. , Saika S. , Yoshizawa S. , Hashizume S. , Horiuchi K. , Okabe N. , Shinozaki T. , Hirayama M.
Published: March 2007

The TD97 strain vaccine virus was prepared from the Tanabe strain measles virus by low‐temperature passages in primary cell cultures and ultraviolet (UV) mutagenesis. The TD97 strain exhibited the following characteristics: highly temperature sensitive, neither multiplying nor forming any plaques at 40°C in Vero cells; genetically stable, maintaining high temperature sensitivity after ten successive passages in CE cells at 30°C or 35°C; and M proteins of this virus about 1 KD slower in mobility in SDS‐PAGE than that of the Tanabe strain. The TD97 strain was further confirmed to be attenuated by an inoculation test into primate brain. In field trials, 752 healthy children were inoculated with a live virus vaccine prepared with this strain, and the following results were obtained: the seroconversion rate was 97% (517/533), and the average HI antibody titer was 25.2. An antibody‐increasing effect was also observed in children who were initially seropositive. In children who seroconverted, the rates of fever were 15.7% (55/351) for 37.5°C or higher and 4.0% (14/351) for 39°C or higher. The rash rate was 7.7% (27/351), and the incidence of local reaction was 5.4% (19/351). The TD97 strain is thus considered to be suitable in use for an attenuated measles vaccine.
Detection of Inoue‐Melnick virus (IMV)‐antibody in Japanese patients with colorectal carcinoma and isolation of IMV from Japanese colorectal carcinoma cell lines
Authors: Nishibe Y. , Inoue Y. K. , Nakamura Y.
Published: March 2007

The antibody prevalence rate of 3 types of IMV in 24 sera of Japanese patients with colorectal adenocarcinoma (mean age 60.8 years) was 83.3% (20/24). Of 20 positive sera, 19 contained type 1 antibody and one contained type 2. In contrast, the seropositivity of IMV in 21 sera of matched control patients (mean age 60.9 years) with non‐colorectal carcinoma such as mammary and lung carcinoma was 23.8% (5/21). The 5 positive sera contained type 1 antibody. The seropositivity to IMV in 79 sera of healthy Japanese over 50 years of age (mean age 62.3 years) was 24.0% (19/79). Again all of the 19 positive‐sera contained type 1 antibody. The seropositivity of IMV in the patient with colorectal adenocarcinoma was significantly higher than in the controls, non‐colorectal carcinoma patients and healthy persons over 50 years of age (P<0.001). Type 1 IMV was isolated from each of 2 recently established Japanese colorectal adenocarcinoma cell lines.
Low prevalence of antibody to hepatitis C virus in North East England
Authors: Brind A. M. , Codd A. A. , Cohen B. J. , Gabriel F. G. , Collins J. D. , James O. F. W. , Bassendine M. F.
Published: March 2007

The prevalence of antibodies to hepatitis C virus (anti‐HCV) was studied in North East England in blood donors, local multiply transfused patients, local high risk individuals, and chronic liver disease patients. Anti‐HCV was detected by enzyme‐linked immunosorbent assay (ELISA) in 2/1120 (0.18%) blood donors; 1/84 chronic renal failure patients on haemodialysis who had received 1,992 units of blood (seroconversion rate of 0.05% per unit transfused), 1/207 cardiac patients 6 months post cardiac surgery transfused with 1,403 units of blood (1 anti‐HCV pre‐operatively, seroconversion rate 0.07%), 40/50 haemophilia A patients treated with commercial factor VIII, and 38/100 intravenous drug users. In addition anti‐HCV was detected by ELISA in 5/35 cryptogenic chronic liver disease patients, 5/5 confirmed by recombinant immunoblot assay (RIBA) (14%); 3/30 patients with autoimmune chronic active hepatitis, 2/3 by RIBA (7%); 2/50 primary biliary cirrhosis patients, 1/2 by RIBA (2%); 0/30 alcoholic cirrhosis patients; and 2/9 patients with hepatocellular carcinoma, 1/2 by RIBA (11%). HCV is uncommon in North East England; it may be implicated in the aetiology of a minority of cases of cryptogenic liver disease and <5% of autoimmune chronic active hepatitis and primary biliary cirrhosis.
Efficient cloning of hepatitis B virus DNA from single‐stranded replicative intermediates and its application to S1 mapping of viral RNA in human liver tissue
Authors: Kajino Kazunori , Ohta Yasuyuki , Miyamura Tatsuo , Saito Izumu
Published: March 2007

An efficient method for cloning subgenomic fragments of the hepatitis B virus (HBV) was developed, utilizing its abundant single‐stranded replicative intermediates. The total genomic DNA obtained from the liver tissue of patients with chronic HBV infection was treated by using the Klenow fragment of E. coli DNA polymerase I without adding any exogenous primers. Single‐stranded replicative intermediates were efficiently converted to double‐stranded linear DNA, one end of which terminated at (or near to) the direct repeat 1 (DR1) sequence of the HBV genome. By screening less than 1,000 recombinants from a DNA library after this treatment, we obtained a subgenomic HBV fragment of 2.0 kilobases. We then analysed HBV RNA in human liver tissue by S1 mapping. It was possible to map HBV RNA only when a DNA probe from the same tissue was used.
Typing and molecular characterization of human papillomaviruses in genital warts from South African women
Authors: Johnson Jennifer E. , Dehaeck Catherine M. C. , Soeters Robbert , Williamson Anna‐Lise
Published: March 2007

Condyloma acuminata from 12 women were examined for the presence of human papilloma‐virus DNA. Six of the women had HPV 6a, two had HPV 11a, one had a new HPV 6 subtype, and one a new HPV 11 subtype. The new HPV 6 subtype could be distinguished from other HPV 6 subtypes on the basis of Pst 1 fragments as well as Hind II fragments. The new HPV 11 subtype had a different Hpa 11 restriction endonuclease pattern. Restriction maps of both new subtypes were constructed. Two of the biopsies did not contain detectable HPV DNA when probed with HPV 6, HPV 11, and HPV16. Biopsies were taken from normal tissue 1 cm away from the condyloma in 11 of the patients. Only one of these normal tissue biopsies was positive for HPV by Southern blot hybridization.
Hepatitis C virus seroprevalence in Italian haemophiliacs injected with virus‐inactivated concentrates: Five year follow‐up and correlation with antibodies to other viruses
Authors: Pistello M. , Ceccherini‐Nelli L. , Cecconi N. , Bendinelli M. , Panicucci F.
Published: March 2007

The overall prevalence of anti‐HCV antibody in a group of 125 haemophiliacs was 62%. Four patients who had never received replacement therapy were anti‐HCV negative. Of the 121 patients injected regularly with commercial concentrates, 76 were already anti‐HCV seropositive in 1985 and remained so throughout the follow‐up. Two patients seroconverted in 1987 without obvious signs or symptoms of hepatitis. Our patients were treated with dry heat‐treated concentrates since 1985 and with wet heat‐ or solvent/detergent‐treated concentrates since 1988. The absence of further seroconversions and of symptoms of acute post‐transfusion non‐A, non‐B hepatitis since 1988 suggest that present virucidal treatments of concentrates are effective in preventing HCV transmission. Anti‐HCV positivity appeared to be unrelated to the type and degree of haemophilia as well as to the presence of antibodies to hepatitis B virus, human immunodeficiency virus type 1, and human herpesvirus type 6.
Human immunodeficiency virus‐1 (HIV‐1) in the vapors of surgical power instruments
Authors: Johnson Gregory K. , Robinson William S.
Published: March 2007

Cool vapors and aerosols produced by several common surgical power instruments and hot smoke plumes generated with electrocautery on known HIV‐1 innoculated blood were gently bubbled through sterile viral culture media. Tissue culture cells were then added and cell infection was detected by the appearance of HIV‐1 P‐24 core antigen assayed by ELISA in the culture medium. HIV‐1 was cultured from cool aerosols and vapors generated by a 30,000 RPM spinning router tip, an instrument similar to the Midas Rex and the Stryker oscillating bone saw. No infectious HIV‐1 was detected in aerosols generated by a Valley Lab electrocautery or with a manual wound irrigation syringe known as a Travenol Uromatic irrigator. We have demonstrated that HIV‐1 can remain viable in cool aerosols generated by certain surgical power tools and this raises the possibility of HIV transmission to medical personnel exposed to aerosols similarly generated during the care of HIV infected patients. Further work is required to determine whether such a risk exists but caution should be exercised by those exposed to aerosols generated during procedures on HIV‐1 infected patients.
Detection of B19 parvovirus in human fetal tissues by in situ hybridisation
Authors: Nascimento J. P. , Hallam N. F. , Mori J. , Field Anne M. , Clewley J. P. , Brown K. E. , Cohen B. J.
Published: March 2007

Evidence of B19 parvovirus infection was sought by in situ hybridisation with biotinylated probes in 65 tissue samples from 32 pregnancies (fetuses, products of conception and/or placentas). Twenty‐seven samples were reactive and the results were confirmed by other methods for B19 virus detection in 22 cases. The other methods used were in situ hybridisation with 3H and 35S labelled probes; dot‐blot hybridisation with biotin and 32P labelled probes; polymerase chain reaction assay; negative stain and thin section electron microscopy; and radioimmunoassay for B19 antigen. The five false positive results by in situ hybridisation with biotinylated probes were considered to be due to non‐specific biotin capture and were more frequent with unfixed samples than with formalin fixed material. It was concluded that while biotinylated probes offered advantages over radioactive probes for detecting B19 DNA by in situ hybridisation, positive findings should be confirmed by other methods.
Restriction site mapping of four genome types of adenovirus types 3 and 7 isolated in South America
Authors: Niel C. , Moraes M. T. B. , Mistchenko A. S. , Leite J. P. G. , Gomes S. A.
Published: March 2007

New genome types of Ad3 and Ad7 were found among adenovirus (Ad) strains isolated from stools of children during epidemiological surveys made in Säo Paulo, Brazil, and Buenos Aires, Argentina. These were characterized by DNA analysis with 11 restriction endonucleases and showed a number of new restriction patterns, notably for BamHI, BcII, BgIII, HindIII, Kpnl, and Smal. Restriction maps of the genome types, named Ad3e1, Ad3e2, Ad3h, and Ad7h, were constructed and compared with those of Ad3p and Ad7p.
A prospective study of post‐transfusion non‐A, non‐B (type C) hepatitis following cardiovascular surgery in Taiwan
Authors: Lee Shou‐Dong , Tsai Yang‐Te , Hwang Shinn‐Jang , Wu Jaw‐Ching , Yung Chien‐Hui , Cheng Kwok‐Kei , Lo Kwang‐Juei
Published: March 2007

In an attempt to investigate the incidence and clinical course of non‐A, non‐B (NANB) hepatitis following blood transfusion in Taiwan, 288 patients who underwent cardiovascular surgery and received blood transfusion were followed prospectively with serum liver aminotransferase levels and viral hepatitis markers for at least six months. None had any past history of liver disease or drug abuse. All blood donors were tested for serum hepatitis B surface antigen and alanine aminotransferase (ALT) (>45U/L). Thirtyseven (12.8%) patients developed PTH. 34 (91.9%) were considered to be cases of NANB hepatitis, 2 (5.4%) were cytomegalovirus hepatitis, and one (2.7%) was caused by Epstein‐Barr virus. No one developed hepatitis B post‐transfusion hepatitis (PTH). Of the 34 NANB PTH patients, 15 (44.1%) were asymptomatic, 16 (47.1%) had clinical symptoms, and 9 (26.5%) had serum total bilirubin levels higher than 2 mg/dl. There was no case of fulminant hepatic failure. Of 26 NANB PTH patients who were followed up for more than one year, 15 (57.7%) still had abnormal serum ALT levels. The incubation period of NANB PTH ranged from 2 to 16 (mean 6.1 ±3.2) weeks. Of the 37 PTH patients, 32 (86.5%) were found to have anti‐HCV seroconversion during one year follow‐up period. NANB PTH is as common in Taiwan as in the United States and Japan, and is demonstrated by this study to be due mostly to HCV.
Characterization of immune complexes containing cytomegalovirus‐specific IgM antibodies following a kidney graft
Authors: Basson Jérôme , Tardy Jean‐Claude , Aymard Michèle
Published: March 2007

In order to demonstrate the viral specificity of IgM‐containing immune complexes (IgM‐CIC) detected by a C1q assay in renal allograft recipients developing a CMV infection, a technique is described allowing: (1) the dissociation of IgM‐CIC by action of an acid buffer, and (2) the characterization of the viral specificity of IgM antibodies released by this treatment. This step was performed by ELISA and Western Blot.

When technique was applied to the follow‐up of a renal allograft recipient developing a recurrent CMV infection within 2 months post‐graft, it was found that the IgM‐CIC detected on the day of the graft were not CMV‐specific, whereas the IgM‐CIC detected during the second month after transplantation contained CMV‐specific IgM antibodies. These CMV‐specific IgM‐CIC were detected as early as the urinary viral excretion.

It was shown by Western Blot analysis that these IgM antibodies reacted with a 45–47 kDa viral polypeptide which is a viral target for specific humoral response at the early phase of CMV infection.
Characterisation of a linear binding site for a monoclonal antibody to hepatitis B core antigen
Authors: Sällberg Matti , Rudén Ulla , Magnius Lars O. , Harthus Hans P. , Noah Michael , Wahren Britta
Published: March 2007

The complete amino acid (aa) sequence of the hepatitis B virus (HBV) core protein (HBcAg), ayw subtype, was synthesized as decapeptides with five overlapping aas. The peptides were tested for reactivity with monoclonal antibodies (mAbs) to HBcAg (35/312, 37/275, and 7/275). All the mAbs specifically inhibited human anti‐HBc by cross competition in assays for anti‐HBc and anti‐HBe.

The mAb 35/312 recognised a peptide covering residues 76–85 of the HBcAg sequence. The other two mAbs did not react specifically with any linear peptide, suggesting discontinuous epitopes for these mAbs. The linear sequence EDPASR at residues 77–82 was found to constitute the epitope for mAb 35/312 when fine mapping the binding site. The most essential aas for mAb 35/312 were found to be the DP at residues 79–80, when peptides were synthesized where the aas at 77–83, were substituted by the other 19 aas. Since the mAb 35/312 inhibits the binding of human anti‐HBc positive sera, which are known to recognise an SDS labile epitope, the sequence 77–82 might be a part of a larger discontinuous epitope. Alternatively the mAb 35/312 blocks the binding of human anti‐HBc by steric hindrance.
Cytotoxicity and cytoadherence of human and murine polymorphonuelear leukocytes against Junin Virus infected targets in the absence of anti‐viral antibody and complement
Authors: Portiansky Enrique Leo , González Pedro Horacio , Laguens Rubén Pedro
Published: March 2007

Human and murine polymorphonuclear leukocytes (PMN) lysed L cells infected with Junin Virus (JV) in an in vitro system free of antiviral antibody and complement. Infected VERO cells proved to be resistant to the cytolytic effect. Murine PMN showed an increased adherence on JV‐infected L cells but did not attach to VERO cells. The opposite was observed with human PMN, which did adhere to infected VERO cells but not to infected L cells. These results indicate that PMN activity may be an early mechanism of defense in JV infection by lysing virus‐infected cells when no immune response has been established, and that cytoadherence is not a necessary step for target lysis.
Significance of IgM anti‐hepatitis D virus (HDV) in chronic HDV infection
Authors: Lau Johnson Y. N. , Smith Heather M. , Chaggar Kanchan , Hansen Lucy J. , Portmann Bernard C. , Alexander Graeme J. M. , Williams Roger
Published: March 2007

Intrahepatic hepatitis D virus (HDV) antigen (HDAg) and serum HDV RNA are excellent markers of active HDV replication but the relation of IgM anti‐HDV to HDV replication and histological activity is less certain. To further elucidate the significance of serum IgM anti‐HDV, 90 paired sera and liver biopsies from 64 patients sero‐positive for total antibody to HDV were analysed for IgM anti‐HDV, intrahepatic HDAg expression, and histological inflammatory activity. IgM anti‐HDV was strongly associated with intrahepatic HDAg expression with a sensitivity of 94.1% but the assay lacked specificity since 14 out of 22 cases negative for intrahepatic HDAg were also positive for IgM anti‐HDV. In 20 patients in whom follow‐up biopsies and paired sera were available, two patients lost intrahepatic HDAg but paired serum remained IgM anti‐HDV positive. Although the presence of serum IgM anti‐HDV correlated significantly with a higher histological inflammatory activity (P = 0.001), there was a considerable overlap with the group seronegative for IgM anti‐HDV, again indicating a poor specificity. This lack of specificity of IgM anti‐HDV for both HDV replication and histological activity indicates that this assay provides no additional information over and above assay for total antibody to HDV.
Reinfection of mice with respiratory syncytial virus
Authors: Graham Barney S. , Bunion Lori A. , Wright Peter F. , Karzon David T.
Published: March 2007

A BALB/c model of respiratory syncytial virus infection has been developed in which hightitered replication occurs in lung, immunological infiltrates in lung can be detected histologically, and illness can be consistently reproduced. The immunodeterminants of RSV reinfection in this system were investigated by rechallenging mice with RSV early (less than 2 months after primary infection) and late (16 to 21 months after primary infection) and correlating illness and titer of RSV isolated from lungs and noses with RSV‐specific serological responses and lung histology. After early rechallenge, RSV was cleared within 24 hours from both nose and lung. After late rechallenge, RSV was isolated at 72 hours from nose in 21/24 mice, but from lung in only 5/24 mice. Isolation of RSV from lung after rechallenge was associated with low RSV‐specific antibody titers measured by ELISA and plaque‐reduction neutralization. The presence of lymphocyte aggregates around the bronchovascular bundles was associated with inability to isolate RSV from lung and lack of illness. The basophilic lymphocytes were small and uniform in size with dense nuclei and a small amount of cytoplasm. These studies demonstrate that nasal and pulmonary reinfection with RSV is possible in mice after late rechallenge. The studies also indicate the potential importance of RSV‐specific antibody in protecting lung from reinfection.
HIV reverse transcriptase inhibiting antibodies detected by a new technique: Relation to p24 and gp41 antibodies, HIV antigenemia and clinical variables
Authors: Neumüller Magnus , Karlsson Anders , Lennerstrand Johan , Källander Clas F. R. , Holmberg Viveca , Långström‐Persson Ulla , Thorstensson Rigmor , Sandström Erik , Gronowitz J. Simon
Published: March 2007

A new assay for HIV reverse transcriptase activity inhibiting antibodies (RTI‐ab) was used for the analysis of a large collection of sera sampled before and after confirmation of HIV infection. In this assay HIV‐RT was preincubated with diluted serum, after which residual RT activity was determined by a technique using a template coupled to macrobeads and125l‐lodo‐deoxyuridine‐triphosphate as the tracer‐substrate. Of the 936 sera analysed, 818 were found positive for RTI‐ab, and 824 were positive in Western blot (Wb). The prevalence of RTI‐ab compared to Wb was therefore 99.3%. The corresponding figure for 930 sera analysed for envelope‐ab, i.e., gp41‐ab, was 823 positive, and of these 930 sera 815 were Wb positive, giving a comparative prevalence of 101%. In contrast, only 678 samples of 993 analyzed for core ab, i.e., p24, were positive, giving a prevalence of 77.0% as 880 of these samples were Wb positive. Thus, RTI‐ab was as prevalent as gp41‐ab, and although the analyses of RTI‐ab amounts in different stages showed decreasing levels in stage IV compared to stages II or III, all of the sera except 1 were found positive in stages III and IV. Further, it was found that both the few RTI‐ab negative samples in stage II and the few RTI‐ab positive samples among Wb negative sera were sampled in connection with seroconversion. The specificity of the RTI‐ab assay was 100% in a test of 200 serum samples from HIV negative blood donors. It was concluded that RTI‐ab analyses can be made highly sensitive and specific and useful for studies of HIV infection.
Prevalence of hepatitis C in tropical communities: The importance of confirmatory assays
Authors: Tibbs C. J. , Palmer S. J. , Coker R. , Clark S. K. , Parsons G. M. , Hojvat S. , Peterson D. , Banatvala J. E.
Published: March 2007

The prevalence of antibody to hepatitis C virus (HCV) was estimated in 3 tropical populations using 2 screening ELlSAs to detect antibody to the c100‐3 antigen and 2 supplementary assays designed to test the specificity of these tests. Two hundred and eighty‐six of 385 (74.2%) sera from Kiribati, 17 of 138 (12.3%) sera from Vanuatu, and 39 of 173 (22.5%) sera from Zaire were reactive in the initial screening assay. The proportion of reactive sera which were also reactive in the second screening ELISA varied between populations (55.1% in Kiribati, 85.1% in Vanuatu, and 39.2% from Zaire). Reactive sera were selected at random for confirmatory testing. Only 3 of 49 (6.12%) of sera from Kiribati and 1 of 14 (4.76%) of sera from Vanuatu positive in the initial ELISA were reactive in the confirmatory assays. The proportion of confirmed positive sera from Zaire was higher 8 of 28 (28.5%). Based on the results of these supplementary assays the estimated prevalence of anti‐HCV in these populations is 4.8% in Kiribati, <1% in Vanuatu, and 6.4% in Zaire. Reliance on a single screening ELISA to estimate the prevalence of anti‐HCV in stored sera from tropical communities may lead to a gross over‐estimate of the true prevalence in these populations.
A randomised controlled trial of recombinant interferon‐γ in chinese patients with chronic hepatitis B virus infection
Authors: Lau Johnson Y. N. , Lai C. L. , Wu P. C. , Chung H. T. , Lok Anna S. F. , Lin H. J.
Published: March 2007

Nineteen Chinese patients with chronic hepatitis B virus (HBV) infection, seropositive for HBV e antigen (HBeAg) and HBV DNA on at least three occasions in 6 months, were randomised to receive either recombinant human interferongamma (rlFNγ) 0.1 mg/m2 intramuscularly thrice weekly for 16 weeks (n = 11) or no anti‐viral therapy (controls, n = 8). Five patients in the treatment group and four patients in the control group had persistently elevated serum alanine aminotransferases (ALT) of over two times the upper limit of normal before entering into the trial.

Rlfnγ had no or minimal inhibitory effect on serum HBV DNA during treatment and no patient developed e‐seroconversion or sustained loss of serum HBV DNA. Hepatitic flare, which occurred in a proportion of patients responding successfully to interferon‐α (IFNα) therapy, was not observed with rlFNγ treatment. Side‐effects included pyrexia and mild headache that showed tachyphylaxis and were well tolerated by all patients. In the control group, one patient with elevated pre‐entry serum ALT lost serum HBV DNA and seroconverted to anti‐HBe. Another patient with elevated ALT lost serum HBV DNA transiently during therapy. In the dose given, rlFNγ was safe but had no apparent anti‐viral effects in Chinese patients with chronic HBV infection.
Prevalence of HBsAg carriers in native and immigrant pregnant female populations in Israel and passive/active vaccination against HBV of newborns at risk
Authors: Bogomolski‐Yahalom V. , Granot E. , Linder N. , Adler R. , Korman S. , Manny N. , Tur‐Kaspa R. , Shouval D.
Published: March 2007

Israel has no official prevention policy at present against perinatal and horizontal transmission of hepatitis B virus (HBV) infection in newborns and children at risk. The present study was designed to assess the prevalence of HBV carrier state in a population of 11,123 pregnant women at term. Among this population (mean age 29.7±5.9),98 women (0.88%) were found to be asymptomatic HBsAg+ carriers, and 97% of these carriers were anti‐HBe+. Evidence for HBV replication, as determined by serum HBV‐DNA, was established in 6.6% of the HBsAg+/anti‐HBe+ population. The HBsAg carrier rate was strongly influenced by religion, continent, and country of birth of the carrier mothers. The highest relative carrier rate was found among women of Moslem origin (4.3%), as compared to Jewish women (0.67%). Most carrier women were born in Israel (56.1%) to mothers who had emigrated from regions with intermediate or high endemicity of HBV, such as North Africa or the Middle East. In these groups, the HBsAg carrier rate ranged between 1.2 and 3.0%. Ninety‐three percent of newborns receiving passive/active vaccination against HBV developed protective levels of anti‐HBs. Finally, evidence for horizontal transmission of HBV was found in 19.3% of 83 non‐vaccinated children in families of HBsAg carriers. The present study therefore establishes HBsAg prevalence rates in specific risk groups of women at term and confirms the need for an official policy on immunization against HBV in Israel. Since over 50% of women at term belong to the defined risk groups, universal active vaccination of the entire newborn population each year is suggested as the most rational and needed policy in Israel.
Further evaluation of a live hepatitis A vaccine in marmosets
Authors: Provost P. J. , Banker F. S. , Wadsworth C. W. , Krah D. L.
Published: March 2007

Live, attenuated F́ hepatitis virus was studied in vivo in Saguinus labiatus marmosets for possible reversion to virulence, for possible establishment of persistent infection and for its capacity as a parenterally administered vaccine to induce immunity to oral infection. Serial transmission of the virus in S. labiatus, using infectious stool extracts for the second and third passages, produced no evidence of reversion of the F́ vaccine virus to virulence. Monitoring for live HAV in stools over a 135‐day period postinoculation of marmosets with the F′ vaccine revealed no evidence of persistent infection. Vaccinated animals were also shown to be resistant to infection on challenge by the oral route as well as by the previously demonstrated parenteral route.
Persistence of HIV‐1 silent infection in seronegative subjects at high risk
Authors: Pezzella Mario , Rosci Maria A. , Miceli Michela , Vonesch Nicoletta , Sturchio Elena , Mirolo Marina , Mannella Emilio
Published: March 2007

Twenty regular sexual partners of HIV‐1 infected subjects, without detectable human irnmunodeficiency virus (HIV‐1) antibody and positive for HIV‐1 genome by in situ hybridization (ISH), were selected and studied longitudinally for 6–36 months to estimate the duration of silent infection. During the follow‐up period, 10 showed atypical Western Blot (WB) patterns. Two seronegative partners seroconverted. Rapid progress t o AIDS was observed in 7 seropositive subjects.
Hepatitis B virus DNA in sera of virus carriers positive exclusively for antibodies to the hepatitis B core antigen
Authors: Luo Kang‐Xian , Zhou Rong , He Chao , Liang Zhi‐Sen , Jiang Shibo
Published: March 2007

The prevalence of serum HBV DNA in individuals positive for anti‐HBc alone was determined by the polymerase chain reaction in two groups with endemic HBV infection from Canton (group A) and Hainan (group B), provinces of China. Twenty‐one out of 294 individuals in group A (7.2%) and 193 out of 1995 in group B (9.7%) were positive for anti‐HBc but negative for other markers of ongoing or past HBV infection. (HBsAg and anti‐HBs). HBV DNA was detected in 6/21 sera in group A (28.6%) and 68/193 in group B (35.2%) in their initial serum specimen. One of the six HBV‐DNA‐positive individuals in group A became negative after 6 months and four of the 58 positive in group B became negative at 4 years of follow‐up. All of the individuals remained positive for anti‐HBc and negative for anti‐HBs, but one of them became positive for HBsAg on follow‐up. None of the anti‐HBc‐ and HBV‐DNA‐positive subjects had symptoms of liver diseases. They were, therefore, defined as chronic asymptomatic HBV carriers with undetectable HBsAg. This type of carrier should be added to the typical HBsAg‐positive carrier, who constitutes about 10–15% of the general Chinese population, to give a more complete estimate of asymptomatic HBV carriers in China.