bluuebunny/crossref_metadata_2025 huggingface dataset:
Typing of human papillomaviruses in cervical intraepithelial neoplasia grade 3 biopsies from cape town
Authors: Williamson Anna‐Lise , Dehaeck Catherine M. C. , Soeters Robbert
Published: March 2007

DNA from 98 cervical intraepithelial neoplasia grade 3 (CIN 3) biopsies was screened, using the Southern blotting hybridization technique, for human papillomavirus (HPV) types 6, 11, 16, 18, 31, 33, and 35. HPV 16 was detected in 16 biopsies (16%), HPV 33 in eight biopsies (8%), HPV 31 in two biopsies (2%), and HPV 18 in one biopsy (1%). One of the biopsies contained both HPV 31 and 18. Six biopsies (6%) contained an HPV type very similar but not identical to HPV 16, and 35 biopsies were positive for HPV, but the Pst 1 restriction fragments were distinct from any of the HPV types used as probes. HPV was not detected in 32% (32/98) of the biopsies screened.
Molecular identification of a novel human rotavirus in relation to subgroup and electropherotype of genomic rna
Authors: Nakagomi Osamu , Oyamada Hajime , Kuroki Satoru , Kobayashi Yasuko , Ohshima Atsushi , Nakagomi Toyoko
Published: March 2007

A total of 41 stool rotavirus specimens collected from children with acute diarrhea at four different locations in Akita Prefecture, Japan, during the peak of the winter diarrhea epidemic in 1988 were analyzed by polyacrylamide gel electrophoresis of viral RNA in conjunction with subgrouping assay. We found that a single strain predominated, with cocirculating strains with less common electropherotypes at a given location, and that two different strains could predominate at geographically close but different locations even during a very limited time of the epidemic season. Furthermore, we isolated a human rotavirus strain (AU125) that was similar to the AU‐1 strain in that it possessed a long RNA pattern yet belonged to subgroup l. Genetic analysis by RNA‐RNA hybridization assay indicated that the AU125 strain was distinct from two previously identified human rotavirus gene groups (genogroups) represented by the Wa strain (subgroup II with long RNA electropherotype) and the DS‐1 strain (subgroup I with short RNA electropherotype), but was very closely related to the AU‐1 strain. These data suggest that the genetic diversity of human rotaviruses may be more extensive than was previously thought.
Regulation by dimethylsulfoxide, insulin, and corticosteroids of hepatitis b virus replication in a transfected human hepatoma cell line
Authors: Gripon Philippe , Diot Christian , Corlu Anne , Guguen‐Guillouzo Christiane
Published: March 2007

A human hepatoblastoma cell clone E4 was obtained by transfection of HepG2 cells with a plasmid DNA containing four tandem copies of hepatitis B virus (HBV) genome. Analysis of both intracellular and extracellular viral DNA revealed that this clone exhibited the main steps of the replication process previously found in normal hepatocyte primary cultures experimentally infected in vitro. Indeed, relaxed‐circular, covalently closed circular, and single‐stranded forms of viral DNA were identified in the cells together with complete virions and immature cores in the medium. Furthermore, the ability of these secreted particles to infect normal human hepatocyte cultures was established. These E4 cells were used to evaluate the effect of various soluble factors on HBV replication. Corticosteroids and, to a greater extent, dimethylsulfoxide (DMSO) increased intracellular viral DNA, whereas insulin reduced it dramatically. Parallel changes in the amounts of viral DNA secreted in the medium were observed. Measurement of the albumin secretion rate indicated that cellular and viral activities could be regulated, at least in part, in a coordinated manner.
Immunity in pekin ducks experimentally and naturally infected with duck hepatitis b virus
Authors: Vickery K. , Freiman J. S. , Kearney R. , Murray S. , Cossart Y. E. , Dixon R. J.
Published: March 2007

The immune response to duck hepatitis B virus (DHBV) had not been elucidated. An assay was therefore established to detect the presence of antibody to DHB surface antigen (anti‐DHBs) in serum of experimentally inoculated and naturally infected ducks. Anti‐DHBs in serum was detected by indirect RIA from the percentage inhibition of binding of rabbit anti‐DHBs to purified DHBsAg. Specificity was confirmed by positive and negative controls, infected and noninfected sera, and a mouse monoclonal antibody to DHB core antigen (anti‐DHBc). Serum and liver samples were tested for DHBV DNA by dot‐blot hybridization assay.

Adult ducks repeatedly inoculated with DHBV remained non‐viraemic but developed anti‐ DHBs. This antibody activity neutralized the infectivity of DHBV, which was experimentally inoculated into 1‐day‐old ducklings. In naturally infected flocks anti‐DHBs was detected in a proportion of noninfected adult ducks as well as 1‐ day‐old hatchlings. Anti‐DHBs activity in hatchlings neutralized the infectivity of experimentally inoculated DHBV.

Pekin ducks can therefore mount a neutralizing antibody response to DHBV, and immunity may be transferred in ovo from dam to offspring.
Adenovirus type 7 infections in children in new South Wales, Australia
Authors: De Silva L. M. , Colditz P. , Wadell G.
Published: March 2007

This study describes our experience of adenovirus type 7 infection in children in New South Wales, Australia. Some aspects of the epidemiology of the infection are also recorded. A community outbreak, a hospital‐ward outbreak, sporadic cases, and data from a centralised registry are described. Results of genome‐type identification using restriction endonucleases are given.

Adenovirus type 7b infection was associated with significant mortality and points to the need, previously expressed, for an adequate vaccine for high‐risk infants. Continued surveillance of adenovirus type 7 infections worldwide is necessary to identify genome types so that appropriate vaccines can be developed.
Tropism and histopathology of the d, b, k, and mm variants of encephalomyocarditis virus
Authors: Cerutis Delie R. , Bruner Richard H. , Thomas Donald C. , Giron David J.
Published: March 2007

Variants of encephalomyocarditis virus (EMCV), which are immunologically indistinguishable by hyperimmune serum, produce different disease syndromes in mice. For instance, in ICR Swiss male mice, EMCV‐B produces no overt illness, EMCV‐MM produces severe neurological signs followed by death, EMCV‐D destroys pancreatic beta cells producing a disease syndrome resembling insulin‐dependent diabetes mellitus, and EMCV‐K is lethal but produces no overt signs of infection. The present study was done to determine the tissue tropism and histopathology of each of these EMCV variants in the ICR Swiss mouse model. The data show the highest concentrations in the following organs: EMCV‐D in the pancreas, EMCV‐B in the pancreas, EMCV‐MM in the cerebrum, and EMCV‐K in the medulla/brainstem. They also show that the pathological lesions produced by each variant correlate well with viral titers.
Validation of a first‐generation epstein‐barr virus vaccine preparation suitable for human use
Authors: Morgan A. J. , Allison A. C. , Finerty S. , Scullion F. T. , Byars N. E. , Epstein M. A.
Published: March 2007

The efficacy of a new vaccine preparation against Epstein‐Barr (EB) virus was investigated in cotton‐top tamarins. The vaccine consists of fast protein liquid chromatography‐purified EB virus membrane antigen glycoprotein of 340 Kd (MA gp340) mixed with a synthetic muramyl dipeptide adjuvant emulsified in squalane containing a pluronic polymer; it is suitable for both scaled‐up batch production and eventual administration to man. Vaccinated tamarins rapidly developed ELISA detectable high titre antibodies to MA gp340, and their sera became strongly EB virus‐neutralising. After challenge with a massive 100% carcinogenic dose of EB virus, the vaccinated tamarins had a strikingly low level of circulating EB virus‐carrying mononuclear cells, in contrast to a control animal, and remained entirely free of tumours. This first‐generation vaccine has thus been validated in experimental animals and the way opened for a phase I human trial.
Serological evidence of dengue fever among refugees, Hargeysa, Somalia
Authors: Botros Boulos A. M. , Watts Douglas M. , Soliman Atef K. , Salib Adel W. , Moussa Mahmoud I. , Mursal H. , Douglas C. , Farah M.
Published: March 2007

Epidemics of a malaria‐like illness affected several thousand residents of the Dam Camp, a refugee camp near Hargeysa in Somalia, during 1985, 1986, and 1987. The disease was characterized by fever, chills, sweats, headache, back and joint pains for as long as 10 days in some patients. Blood smears from acutely ill patients were negative for malaria. Of 28 acute and 10 convalescent sera tested by the indirect fluorescent antibody (IFA) and by the hemagglutination inhibition (HI) tests, all were negative for antibody to Rift Valley fever, Crimean‐Congo hemorrhagic fever, Sindbis, Chikungunya, yellow fever, and Zika viruses. However, antibody reactive to dengue 2 virus was detected by the IFA test in 39% (15/38), and 11 of 29 (38%) of the same sera were antibody positive by the HI test. Also, IgG antibody reactive to dengue 2 was demonstrated in 60% (17/28) of the same sera by the enzyme immunoassay (EIA), and 14% (4/28) were positive for IgM antibody. Of ten patients for which acute and convalescent sera were available, two developed four fold or greater rises in antibody titer evidencing infection. These data suggested that dengue virus may have been the cause of the epidemic among the Dam Camp refugees.
Detection of human cytomegalovirus immediate early antigen in leukocytes as a marker of viremia in immunocompromised patients
Authors: Grazia Revello M. , Percivalle Elena , Zavattoni Maurizio , Parea Maurizio , Grossi Paolo , Gerna Giuseppe
Published: March 2007

Peripheral blood polymorphonuclear (PMN) cells from 35 immunocompromised patients (22 heart transplant recipients and 13 AIDS patients) and four normal subjects were tested for the presence of human cytomegalovirus (HCMV) immediate early antigen (IEA) (antigenemia) by indirect immunofluorescence (IFA) and IEA‐specific monoclonal antibodies (MAb). PMN samples were tested in parallel for HCMV isolation (viremia) by using MAb to viral early antigens (EA) and the IFA technique 24–48 hr after inoculation onto human fibroblast monolayers. HCMV was isolated from 26 of 83 PMN samples examined: of these, 25 were also positive for HCMV IEA (96% sensitivity). Seven additional PMN samples negative for viral isolation resulted IEA‐positive (87.7% specificity). Six of the seven discordant samples were taken from four patients during ganciclovir treatment. The transitory dissociation between positive HCMV antigenemia and negative viremia during antiviral treatment was followed, at the end of the therapy, either by virus clearance and disappearance of IEA‐positive PMNs (one patient) or by reappearance of viremia (three patients). Among concordant positive samples, a significant correlation was observed between the number of IEA‐positive PMN leukocytes and EA‐positive nuclei of infected fibroblasts, when the same number of PMNs were used for both tests.
Complement‐Independent neutralising monoclonal antibody with differential reactivity for strains of human cytomegalovirus
Authors: Baboonian C. , Blake K. , Booth J. C. , Wiblin C. N.
Published: March 2007

A mouse monoclonal antibody with complement‐independent neutralising activity against cytomegalovirus (CMV) and reactive with the 86 kilodalton (kDa) viral glycoprotein H is described. Neutralisation tests against a range of different strains of CMV showed significant crossreactivity, but clear differences were evident between the two prototype viruses AD169 and Davis, and particularly between AD169 and several low‐passage recent clinical isolates; CMV present in urine was neutralised weakly if at all.
Comparative assessment of antiretrovirals in human monocyte‐macrophages and lymphoid cell lines acutely and chronically infected with the human immunodeficiency virus
Authors: Crowe Suzanne M. , McGrath Michael S. , Elbeik Tarek , Kirihara Jean , Mills John
Published: March 2007

We compared the efficacy of 3 antiretrovirals in cultured human peripheral blood monocyte‐derived macrophages and lymphoid cells infected with the human immunodeficiency virus (HIV). Zidovudine (>0.01 μg/ml) orfoscarnet (100 μg/ml) consistently inhibited HIV replication (p24 antigen production) in acutely infected macrophages by more than 90%; alpha interferon, (1,000 units/ml) inhibited HIV replication by 88–99%. Drug efficacy was equal in lymphoid cells and monocyte‐derived macrophages. However, these antiretrovirals even in high concentration only minimally inhibited chronic, established HIV infection: while zidovudine (0.01 μg/ml) inhibited infection by over 90% in acutely infected macrophages and lymphoid cells, 50 μg/ml of this antiretroviral produced only 19–55% inhibition of HIV in chronically infected cells. Slot‐blot analysis of HIV RNA was concordant with the p24 antigen data. Acutely infected macrophages are as susceptible to the inhibitory effects of antiretrovirals as lymphoid cells; however, chronically infected macrophages (which probably constitute the main in vivo reservoir for HIV) are several orders of magnitude more resistant.
Mutants of the hepatitis b virus surface antigen that define some antigenically essential residues in the immunodominant a region
Authors: Ashton‐Rickardt Philip G. , Murray Kenneth
Published: March 2007

The nature of the immunodominant a region of hepatitis B virus surface antigen (HBsAg) has been examined by mutation of specific amino acid residues. Proline 142 was required for the exhibition of full antigenicity. Replacement of cysteines 124 and 147 by serines drastically reduced or eliminated reactivity with antibodies to HBsAg, which implicates these two residues in stabilising the conformation of the antigen. The a region of HBsAg has also been shown to influence both the immunoreactivity of the adjacent subtype antigenic region, despite being immunologically distinct from it, and the ability of the antigen to interact with a subtype‐specific monoclonal antibody. These results emphasise the importance of the polypeptide region between cysteine residues 124 and 147 in determining a antigenicity, as well as manifestation of the subtype of HBsAg, and reinforce the view that these are conformational epitopes.
Formalin inactivated junin virus: Immunogenicity and protection assays
Authors: Videla Cristina , Carballal Guadalupe , Remorini Patricia , Torre José La
Published: March 2007

The aim of this study was to determine if Junin virus inactivated with formalin (FA) was immunogenic and able to elicit a protective response in the guinea pig. The XJ‐Clone 3 strain of Junin virus grown in Vero cells was exposed to FA at 0°C. The following inactivated antigens were prepared: A1, 0.1% FA for 50 hr; A2, 0.1% FA for 50 hr followed by concentration with polyethylene glycol (PEG); B1, 0.05% FA for 70 hr; B2, 0.05% FA for 70 hr plus PEG concentration; C, 0.1% FA for 50 hr followed by ultracentrifugation and purification by sucrose gradient. No residual infectivity was detected in any inactivated antigen either after two passages in newborn mice or by coculture with Vero cells.

After immunization, high‐neutralizing and low‐immunofluorescent antibody titers were obtained in adult mice and guinea pigs, thus showing that antigenicity was preserved. However, in spite of the presence of neutralizing antibodies, guinea pigs gained no protection against challenge with the highly pathogenic XJ strain.

These results suggest that antibody amount and/or quality may be inadequate; alternatively, mechanisms other than humoral immunity, such as cellular response, not elicited by inactivated antigens may be essential for protection against Junin virus.
Efficiency of the polymerase chain reaction for the detection of human immunodeficiency virus type (HIV‐1) DNA in the lymphocytes of infected persons: Comparison to antigen‐enzyme‐linked immunosorbent assay and virus isolation
Authors: Stoeckl Elisabeth , Barrett Noel , Heinz Franz X. , Banekovich Michael , Stingl Georg , Guggenberger Klaus , Dorner Friedrich , Kunz Christian
Published: March 2007

Seventy‐one human immunodeficiency virus type (HIV‐1)‐positive patients were investigated by polymerase chain reaction (PCR), virus isolation, and antigen detection for the existence of HIV in blood. The identification of HIV DNA by PCR, using three different pairs of primers, yielded a clearly higher detection rate (86%) than with two primer pairs (75%) and was far more sensitive than virus isolation (45%) and antigen ELISA (14%). The PCR‐negative results were clearly correlated to asymptomatic clinical stages. However, there was a limited correlation between the clinical stage of disease and the amount of HIV DNA that could be detected in equal numbers of CD4+ cells from different patients, which might be due to their treatment with azido‐thymidine (AZT).
Pig erythrocyte ghost cells used for concentration of enteric viruses from experimentally contaminated clinical specimens
Authors: Armon R. , Neeman I. , Kott Y.
Published: March 2007

A simple, rapid and efficient procedure of virus concentration from urine, cerebrospinal fluid (CSF), and feces was developed. Pig erythrocyte ghost cells were used to adsorb and elute such viruses as poliovirus LSC‐1, echovirus 6, and human rotavirus (clinical isolate). In urine and CSF, the adsorption efficiency range was 80–100% and elution was from 85% to >100%. In addition, poliovirus LSC‐1 was used as an experimental model to examine this procedure under various clinical conditions, such as calcium, glucose, amino acids, and urea at high concentrations. These were added to normal urine specimens to simulate pathological conditions. The results suggest that pig erythrocyte ghost cells are an efficient matrix for adsorption and elution of enteric viruses found in clinical specimens of urine, CSF, and feces. This method might be useful in virus concentration from clinical specimens and for preparative microscopy and other clinical laboratory methods that require subsequent virus concentration.
Preliminary evidence that azidothymidine does not affect hepatitis B virus replication in acquired immunodeficiency syndrome (AIDS) patients
Authors: Farraye Francis A. , Mamish Diaa M. , Zeldis Jerome B.
Published: March 2007

The effect of azidothymidine (AZT) on hepatitis B virus (HBV) replication was determined in three patients with the acquired immunodeficiency syndrome (AIDS). Serum viral DNA was present, and its concentration either remained the same or increased in two patients. Since AIDS patients may be infected with a variety of viral agents in addition to the human immunodeficiency virus (HIV), the effects of the antiviral agents and biological modifiers on other common viral infections should also be determined in developing new approaches to HIV infection. Our results give preliminary evidence that AZT does not affect HBV viral replication in vivo.
Improved Sensitivity of Detection by Avidin—Biotin Complex (ABC) Immunocytochemistry in Epstein‐Barr Virus Serology
Authors: Se Thoe S. Y. , Sam C. K. , Cheng H. M. , Prasad U.
Published: March 2007

Serum antibodies against Epstein‐Barr virus (EBV)‐determined antigens have traditionally been titrated by the indirect immunofluorescence (IIF) technique. The avidin—biotin complex (ABC) immunocytochemical technique was used to determine the serum levels of IgA against EBV viral capsid antigen (IgA/VCA) and IgA against EBV early antigen (IgA/EA) in sera of 106 nasopharyngeal carcinoma (NPC) patients prior to treatment and 100 normal individuals. The sensitivity of the ABC technique is enhanced by an amplification of the antigen—antibody reaction, which involves the binding of the enzyme‐linked ABC to the second biotinylated antibody. There was a good correlation (r = 0.9988) between ABC and IIF‐determined IgA/VCA‐positive titres, with the ABC technique being more sensitive than IIF in the detection of IgA/VCA in NPC sera: 94% (99/106) and 76% (80/106), respectively. The frequency of IgA/EA reactivity in NPC sera was also markedly increased by immunodetection with the ABC technique as compared with IIF technique: 63% (69/106) and 28% (30/106) respectively. Both the immunocytochemical techniques were equally specific in discriminating between elevated serum titres of IgA/VCA and IgA/EA in NPC sera from normal human sera.
Herpes simplex virus infections in male and female mice following pinna inoculation: Responses to primary infection and artificially induced recurrent disease
Authors: Blondeau Joseph M. , Embil Juan A. , Sandra McFarlane E.
Published: March 2007

The pathogenesis of herpes simplex virus (HSV) infection is now well understood. After humans or experimental animals recover from primary infection, the virus remains latent in sensory ganglia of the peripheral nervous system. Latency, for the most part, remains unresolved, and elucidating the mechanisms involved with latency has proved difficult. The requirement for unravelling HSV latency is the availability of a reproducible animal model. Previously, the mouse ear model has been extensively characterized; however, many studies using this model have involved female mice only despite evidence that recurrent HSV infection in humans may vary by gender. We inoculated male and female mice subcutaneously in the pinna with varying amounts of one of four strains of HSV and monitored the mice for signs of primary infection. Following recovery from primary infection, mice were induced to develop recurrent disease. In addition, we attempted to isolate virus from dorsal root ganglia of mice suspected of harboring latent virus. There were no differences in the response of male and female mice to either primary infection or artificially induced recurrent disease when inoculated with the same virus. Differences were noted when female mice were inoculated with different strains of virus.
In vivo administration of monoclonal antibody to the NK 1.1 antigen of natural killer cells: Effect on acute murine cytomegalovirus infection
Authors: Shanley John D.
Published: March 2007

Monoclonal antibody to the NK 1.1 antigen, found on the natural killer cells of a number of strains of mice, specifically suppresses NK cell function when given in vivo. Using this monoclonal antibody, we have examined the effects of specific suppression of natural killer (NK) cells in vivo on acute murine cytomegalovirus (MCMV) infection in C57BL/10ScN mice. Administration of antibody to NK 1.1 substantially lowered the resistance of C57BL/10ScN mice to lethal virus challenge. In addition, antibody administration prior to intraperitoneal infection significantly increased MCMV replication in salivary glands, lungs, and spleens. In C3H/HeN mice, a strain that lacks the NK 1.1 antigen, antibody to NK 1.1 had no effect on virus replication or lethal infection. Thus, in vivo administration of monoclonal antibody to NK 1.1 alters the course of acute MCMV infection. These findings further substantiate the role of NK cells in defense against acute MCMV infection.
Nested polymerase chain reaction assay for the detection of cytomegalovirus overcomes false positives caused by contamination with fragmented DNA
Authors: Porter‐Jordan Kathleen , Rosenberg Eric I. , Keiser John F. , Gross Jeffrey D. , Ross Allan M. , Nasim Suhail , Garrett Carleton T.
Published: March 2007

The polymerase chain reaction (PCR) technique offers a promising alternative to tissue culture for the rapid and sensitive detection of cytomegalovirus (CMV) infection. However, high levels of background amplification detected in samples containing water but no DNA make interpretation of borderline positive samples extremely difficult and reduce the sensitivity of the assay. The signal from amplification of water or positive samples can be eliminated by DNase treatment, but not by filtration through anisotropic membrane, autoclaving, or ultraviolet irradiation. A lag time of 10 to 12 cycles is observed before the reactions with water will show product formation by liquid hybridization detection. The use of nested PCR eliminates the background and, in serial dilutions of a positive sample, shows a 500‐ to 1000‐fold increase in sensitivity by liquid hybridization detection. We suggest that the background signal is arising from small fragments of DNA, which may be produced by autoclaving viral culture material. Such fragments would escape filtration, and overlapping fragments of DNA can prime one another to form complete mosaic sequences that will then amplify. Nested PCR, appropriately controlled for the number of cycles at each step, should successfully overcome such false positives caused by fragmented DNA, no matter if the contamination occurs at the collection site, in processing, or at the facility performing the test.
Detection of human cytomegalovirus early and late antigen and DNA production in cell culture and the effects of dimethyl sulfoxide, dexamethasone, and DNA inhibitors on early antigen induction
Authors: Li S. B. , Fong C. K. Y.
Published: March 2007

Recently a conventional method for the laboratory diagnosis of human cytomegalovirus (HCMV) infection was improved by using centrifugation culture to enhance viral adsorption and by detecting HCMV early antigen and DNA. Comparison of the sensitivity of three rapid methods using commercial diagnostic reagents for the detection of HCMV early antigen (EA), late antigen (LA), and DNA was quantitatively evaluated in centrifugation cultures of human fibroblast cells (MRC‐5) infected with HCMV. HCMV‐EA was first detected 4 hours after infection, and the number of antigen‐positive cells increased rapidly thereafter. Using biotinylated DNA probe, viral DNA was first detected 12 hours postinfection; the number of DNA‐positive cells increased slowly. HCMV‐LA was first seen 48 hours postinfection, and the number of LA‐positive cells also increased thereafter. Thus detection of HCMV‐EA was the most rapid and sensitive method for HCMV diagnosis.

Several chemical compounds have been used to enhance HCMV replication. The effect of dimethyl sulfoxide (DMSO), dexamethasone (DEX), 5‐bromo‐2‐deoxyuridine (BrdU), 5‐fluoro‐deoxyuridine (FdU), and cytosine arabinoside (Ara‐C) on HCMV‐EA induction was evaluated in centrifugation cultures of MRC‐5 cells infected with HCMV. Infected cells treated with 1% DMSO alone or with DMSO plus DEX (10−5 M) have been shown to increase the number of HCMV‐EA—positive cells three‐ to fivefold over the untreated control cultures. The enhancing effects of Ara‐C, BrdU, and BrdU plus FdU were demonstrated only occasionally.
Human papillomavirus types in anogenital warts of children
Authors: Gibson Patricia E. , Gardner Sylvia D. , Best Susan J.
Published: March 2007

Tissue from anogenital warts of 25 children, 10 of whom were suspected of being victims of sexual abuse, was investigated by dot blot and Southern blot techniques for human papillomavirus (HPV) types. HPV DNA was detected in 22 children, two of whom had double infections. The genital HPV types 6 and/or 11 were detected in 20 children, and in three children other HPV types were found. One had HPV 18 (as well as 11); in a second child a possible skin type, HPV 2, was detected; and the third child was infected with an unidentified type.

In three cases genital wart material was available from one of the parents, and in all three the HPV type was the same as that of the child. For nine other children one or both parents were reported to have genital warts. The source of infection appeared to be the adult genital tract, but sexual contact might not be the only means of transmission.
Detection of hepatitis B viral DNA by polymerase chain reaction in patients with hepatitis B surface antigen
Authors: Shih Ling‐Na , Sheu Jin‐Chuan , Wang Jin‐Town , Huang Guan‐Tarn , Chen Juei‐Suei , Sung Juei‐Low , Wang Teh‐Hong , Chen Ding‐Shinn
Published: March 2007

Hepatitis B virus (HBV) DNA was assayed using the polymerase chain reaction in serum samples of 116 hepatitis B surface antigen (HBsAg) carriers, including 30 positive for hepatitis B e antigen (HBeAg) and 86 negative for HBeAg. In the HBeAg‐positive group, all were positive for HBV DNA. In the HBeAg‐negative group, 80.2% were positive for HBV DNA (80.0% in the healthy carrier group, 90.0% in the chronic active liver disease group, and 69.2% in patients with cirrhosis). This study indicated that every HBeAg‐positive carrier as well as the majority of HBeAg‐negative carriers were infectious and, in the latter group, that viral replication is most active in patients with chronic active liver disease.
Diagnostic significance of IgM antibody to hepatitis delta virus in fulminant hepatitis B
Authors: Tassopoulos Nicolaos C. , Koutelou Maria G. , Macagno Silvina , Zorbas Panayiotis , Rizzetto Mario
Published: March 2007

The prevalence of hepatitis delta virus (HDV) infection was studied in 25 adult patients with fulminant hepatitis who were admitted consecutively to our unit from February, 1986, to September, 1988. Enzyme and radioimmunoassays were used for the detection of serological markers of HAV, HBV, and HDV (HDAg, IgM anti‐HD, total [IgG] anti‐HD) infections. Two hundred twenty‐nine serum samples (three to 19 samples/patient) were tested for serological markers of HDV infection. Of the 25 patients, 17 (68%) were HBsAg‐positive, and the remaining eight (32%) were HBsAg‐negative on admission to the hospital. All patients were seropositive for IgM anti‐HBc. Serological markers of HDV infection were detected in 13 (52%) of the 25 patients. In particular, HDV infection was observed in nine (53%) of the 17 HBsAg‐positive and in four (50%) of the eight HBsAg‐negative patients with type B fulminant hepatitis. Survival was 16.7% for patients with hepatitis B and 57.8% for patients with B and D coinfection. Coinfections were responsible for fulminant hepatitis in 100% of drug addicts and 40% in patients who were not drug addicts. All patients with HBV/HDV coinfections became seropositive for IgM anti‐HD. The results show that HDV infection has a significant role (52%) in type B fulminant hepatitis in an area with a moderate prevalence of HBV infections, that it should be tested in cases with early clearance of HBsAg, and that it does not seem to be accompanied by a high fatality rate. Testing for IgM anti‐HD is mandatory for accurate evaluation of the incidence of HBV/HDV coinfections in patients with fulminant hepatitis.
Blocking‐ELISA for detection of specific antibodies to the glycoproteins of the human immunodeficiency virus type 1 (HIV‐1)
Authors: Tardy J. C. , Basson J. , Douglas A. , Aymard M.
Published: March 2007

A blocking ELISA was developed to confirm the specificity of screening tests for anti‐HIV‐1 antibodies. A murine monoclonal antibody (McAb) raised against recombinant gp 160 was used in combination with a commercial technique (ELAVIA‐1). After determining the optimal experimental conditions, the assay was applied to 92 samples presenting different reactivities by Western blot (WB) analysis. All the sera containing antibodies to gp160/gp120 (53) were positive in our assay. The six patients who sero converted showed a low positivity by ELAVIA‐1 (optical density near the cutoff value) reacted by blocking‐ELAVIA‐1 with an McAb binding inhibition greater than 85%. By contrast, negative samples (29) and specimens that exhibited reactivity only against gag‐proteins (10) were not detected (McAb binding inhibition smaller than 15%).

This sensitive and specific blocking‐ELAVIA‐1 represents a convenient alternative to WB as a confirmatory test. The technique is time‐saving and inexpensive and can easily be integrated with a screening test for diagnostic or epidemiologic studies on HIV‐1 infection.
Inhibition of rate of tumour growth in rodent species by inoculation of herpesviruses and encephalomyocarditis virus
Authors: Cowan M. , Davies J. , Brookes K. , Billstrom M. , McLeish P. , Buchan A. , Skinner G. R. B.
Published: March 2007

Inoculation of herpesviruses and encephalomyocarditis virus into subcutaneous tumours in hamsters and mice reduced the rate of tumour growth compared to untreated tumours or secondary tumours which had arisen following surgical excision of the primary tumour; in addition, survival times were increased in animals whose tumours were inoculated with virus. It is suggested that the role of virus in the modification of tumour growth merits further exploration.
Detection of measles virus genomic sequences in sspe brain tissue by the polymerase chain reaction
Authors: Godec Mark S. , Asher David M. , Swoveland Peggy T. , Eldadah Zayd A. , Feinstone Stephen M. , Goldfarb Lev G. , Gibbs C. J. , Carleton Gajdusek D.
Published: March 2007

The polymerase chain reaction (PCR) was modified to detect RNA genomic sequences by generating cDNA copies of those sequences as a preliminary step. Oligonucleotide primer pairs complementary to sequences in each of the five major structural protein genes of the measles virus (nucleocapsid protein, phosphoprotein, matrix protein, fusion protein, and hemagglutinin protein) were synthesized. PCR products were tentatively identified by visualization of bands of the appropriate size by ethidium bromide staining after gel electrophoresis, and identity was confirmed by subsequent restriction enzyme cleavage of the products at predetermined sites to yield fragments of predicted size. This method successfully amplified 400–500 base regions from each of these five genes in RNA extracts of wild measles virus cultured in Vero cells and in RNA extracted from most of the SSPE brain tissues tested, but not in RNA from any control brain tissues. Measles virus genome was detected in SSPE brain tissues stored frozen for as long as 27 years and formalin‐fixed paraffin‐embedded subacute sclerosing panencephalitis (SSPE) brain tissues as old as 9 years. This method provides a simple, rapid and highly sensitive means of detecting and identifying sequences of RNA genomes by PCR. The success of this method in detecting measles virus in SSPE brain tissues suggest that PCR is appropriate to investigate the possible presence of RNA viruses in other neurological disorders of unknown etiology.
Case report: Electron microscopic detection of human parvovirus (B19) in a patient with HIV infection
Authors: Chrystie Ian L. , Almeida June D. , Welch Jan
Published: March 2007

We report the findings on an HIV‐positive patient found to be infected with human parvovirus B19. In a comparative study of sequential sera, direct electron microscopy (EM) had the same sensitivity as DNA hydridisation for the detection of the virus. EM did not require specific reagents and also yielded information on the state of the virus; i.e., whether random or complexed, and the type of antibody involved. The presence of parvovirus complicated the judgement as to drug treatment. This case also highlighted the importance of diagnosing the presence of opportunistic viral infections that have no, or lowgrade, pathogenic activity.
Infection enhancement of influenza A H1 subtype viruses in macrophage‐like P388D1 cells by cross‐reactive antibodies
Authors: Ochiai H. , Kurokawa M. , Kuroki Y. , Niwayama S.
Published: March 2007

The contribution of cross‐reactive hemagglutination inhibition (HI) antibodies to infection enhancement of influenza A H1 subtype NWS virus and two antigenic drift strains was investigated in a macrophage‐like cell line P388D1. When P388D1 cells, previously treated with neuraminidase (NA) to remove the viral receptors, were infected with NWS virus exposed to rabbit antiviral immunoglobulin (IgG) showing various levels of cross‐HI titers, virus yields were enhanced in the presence of a subneutralizing antibody, depending on their cross‐HI titers. By flow cytometric analysis using a fluorescein isothiocyanate (FITC)‐labeled NWS virus, the efficiency of attachment of virus‐rabbit IgG complexes to Fc receptors on NA‐treated cells showed close correlation with its cross‐HI titer. These data suggest that cross‐reactive HI antibodies could contribute to infection enhancement through the formation of potent infectious immune complexes with drift strains to mediate virus infection via Fc receptor uptake. Two monoclonal antibodies (mAB) in mouse IgG subclasses IgG1 and IgG2a showing strain‐specific or cross‐reactive HI activity were tested for their infection enhancement characteristics. A strain‐specific mAB enhanced infection of homologous NWS virus, but not that of two other drift strains in either antibody dilution. In contrast, a cross‐reactive mAB caused infection enhancement of all three virus strains in the presence of the subneutralizing antibody. This indicates that cross‐reactivity, but not the IgG subclass, acts as an enhancing factor to this phenomenon. The antibody, with the same specificity as cross‐reactive mAB, was detected semiquantitatively by competitive enzyme‐linked immunosorbent assay (ELISA) with results almost consistent with cross‐HI titers of polyclonal rabbit antiviral IgGs. These data suggest that the antibody detected by this assay might be one of the potent antibodies governing cross‐HI activity as a whole antibody and causing infection enhancement of drift strains.
Local antibody production and respiratory syncytial virus infection in children with leukaemia
Authors: Taylor C. E. , Craft A. W. , Kernahan J. , Millman R. , Reid M. M. , Scott R. , Toms G. L.
Published: March 2007

Children undergoing therapy for acute lymphoblastic leukaemia (ALL) are at increased risk of severe viral respiratory infection, and some find it difficult to terminate virus secretion. This increased severity may result from a defect in the mucosal immune response. To test this hypothesis, nasal immunoglobulin secretion and specific antiviral antibody responses to infection with respiratory syncytial (RS) virus in children with ALL have been compared with those in a normal age‐matched comparison group.

Children with leukaemia secreted normal levels of IgA and slightly raised lgM levels. lgG levels were depressed. Following RS virus infection, the majority of children with leukaemia secreted normal amounts of lgA and lgG nasal antibody and successfully cleared the virus. However, three of the 13 children studied made poor or undetectable nasal antibody responses, which correlated with their inability to clear the virus.
Clonal origin of mammalian hepatitis B virus‐related hepatocellular carcinoma
Authors: Shimoda Atsushi , Kaneko Shuichi , Uchijima Masato , Unoura Masashi , Hattori Nobu , Kobayashi Kenichi , Murakami Seishi
Published: March 2007

The clonal origins of 20 multifocal hepatocellular carcinomas (HCCs) in four woodchucks were analyzed by the Southern blot hybridization technique. The woodchucks were divided into two groups according to the morphological classification of multifocal tumors: 1) three woodchucks had multifocal tumors that were widely separated and similar in size, which suggests a multiclonal origin of the tumors; and 2) one woodchuck had ten small multifocal tumors surrounding two large main tumors, which indicated intrahepatic metastasis from an original tumor. Results from the first group demonstrated that the number of integrated woodchuck hepatitis virus (WHV) DNAs differed from tumor to tumor, and none of the bands were the same size. In the second group, eight of the ten small tumors surrounding the two large tumors showed the same pattern of WHV DNA integration. One demonstrated an additional band and also shared the same bands with the other tumors, and one small tumor had a different pattern of integration from the others. It was concluded that the clone dissimilarity demonstrated by hybridization patterns does not necessarily mean that HCCs originate independently from different clones, because genetic changes may occur after or at the time of metastasis.
Alterations in the cellular phenotype induced by herpes simplex viruses
Authors: Galloway Denise A. , McDougall James K.
Published: March 2007

Numerous studies have shown that herpes simplex virus types 1 and 2 (HSV‐1, HSV‐2) are able to transform the morphological phenotype of rodent cells. Unlike other DNA tumor viruses the transformed cells did not consistently retain of express a given set of viral genes. In fact, transformation could be obtained using fragments of viral DNA that did not wholly encode viral proteins. Of interest within the transforming fragments were sequences which could assume a secondary structure like that of insertion elements. The failure to detect viral DNA in transformed cells led to the hit‐and‐run hypothesis of HSV transformation. The mechanism by which HSV induces transformation is not understood. Various lines of investigation have shown that HSV is able to cause mutations—both point mutations and gene rearrangements. HSV is also able to induce gene amplification, particularly of sequences harboring an origin of replication such as SV40 or papillomaviruses. Other experiments have shown that HSV can activate the expression of endogenous type C retroviruses. More broadly, HSV has been shown to activate cellular transcription or to switch on the synthesis of host cell proteins not normally expressed in untransformed cells. the failure to detect viral DNA in a high proportion of human anogenital tumors made it difficult to implicate HSV in the etiology of those neoplasias, but it is consistent, however, with the observations on the mode of HSV transformation in vitro, and suggests that HSV could be involved in a multistage process of oncogenic transformation.
Key issues in the selection of an expression system for vaccine antigens
Authors: Ellis Ronald W. , Gerety Robert J.
Published: March 2007

Three criteria by which the appropriate host cell is chosen for the expression of a recombinant‐derived vaccine antigen are efficacy, safety, and scale‐up. Efficacy for a vaccine antigen refers to the ability of the host cell to produce a vaccine antigen capable of eliciting a protective immune response. A concern for safety of a vaccine antigen relates to residual DNA in the final product, especially when derived from continuous mammalian cell lines as opposed to microbial cells. Since tens (or hundreds) of millions of doses of a widely used vaccine might be injected into healthy infants and young children during the lifetime of the product, safety is a critical issue, such that the use of a microbial expression system might be preferable to the use of a continuous cell line in certain circumstances.
Diminished response to recombinant hepatitis B vaccine in homosexual men with HIV antibody: An indicator of poor prognosis
Authors: Loke Richard H. T. , Murray‐Lyon Iain M. , Coleman John C. , Evans Brian A. , Zuckerman Arie J.
Published: March 2007

Three doses of a recombinant DNA HBV vaccine (MSD) were given to healthy male homosexuals. Seventy‐eight out of 104 (77.6%) participants had detectable antibody (anti‐HBs) two months after the third dose. Seroconversion occurred in only 9 out of 27 subjects (33.3%) who were anti‐HIV positive compared with 69 out of 77 (89.6%) who were negative (χ2 = 30.8; P < .001). Fifteen of the 18 anti‐HIV positive who did not mount an antibody response to the hepatitis B vaccine (anti‐HBs) later progressed to persistent generalised lymphadenopathy syndrome (5), AIDS‐related complex (5), and AIDS (5). Only one of the nine anti‐HIV positive anti‐HBs responders developed PGL (χ2 = 10.14; P < .005).

Our results show that anti‐HIV positive homosexuals are poor responders to the recombinant hepatitis B vaccine and anti‐HIV positive non‐responders are more likely to develop clinically apparent HIV infection.
Characterization of a nucleic acid probe for the diagnosis of human coronavirus 229E infections
Authors: Myint S. , Harmsen D. , Raabe T. , Siddell S. G.
Published: March 2007

A cDNA copy of the HCV229E nucleocapsid protein gene was isolated and characterized. Sequence analysis predicts a nucleocapsid polypeptide of 389 amino acids with a molecular weight (mol. wt.) of 43,450. Single strand RNA probes derived from the cDNA copy hybridize specifically to HCV229E RNA and approximately 50 pg of intracellular viral RNA can be readily detected. The application of nucleic acid hybridization as a routine procedure for the diagnosis of HCV229E infection is discussed.
Hepatitis B virus DNA patterns in the liver of children with chronic hepatitis B
Authors: Bartolomé Javier , Moraleda Gloria , Moreno Mercedes Ruiz , Porres Juan Carlos , Carreño Vicente
Published: March 2007

The patterns of hepatitis B virus DNA (HBV‐DNA) expression were studied in 2 sequential liver biopsies from 26 children (18 treated with interferon and 8 controls) with chronic hepatitis B. In the basal biopsy replicative forms of HBV‐DNA were detected in all of the samples and integrated viral DNA was present in 1 case. At the end of the study, 8 children had lost serum HBV‐DNA although 2 of the children were still HBeAg positive. (Six had been treated with interferon.) In all of the cases, HBV‐DNA was not detectable in the final biopsy. For the rest of the patients, HBV‐DNA was positive in serum and all of them had replicative forms of HBV‐DNA in the second liver sample. None of the patients lost hepatitis B surface antigen (HBsAg). Peripheral blood mononuclear cells (PBMC) from these patients were studied. HBV‐DNA was not found in the PBMC of the 8 children without serum HBV‐DNA, and HBV‐DNA was detected in the PBMC of 5/12 patients with serum HBV‐DNA.

In conclusion, HBV‐DNA disappeared from the biopsies of children who lost circulating HBV‐DNA, although some of the patients were still HBeAg positive. This result implies that the detection of HBV‐DNA in liver is important in order to assess the efficacy of the antiviral therapy. On the other hand, HBsAg remained positive in all children at the end of the study although HBV‐DNA was not detected in serum, liver, and PBMC by the conventional hybridization techniques.
Detection of respiratory syncytial virus antigen after seventy‐two hours of culture
Authors: Mézière Annyvonne , Mollat Claudine , Lapied Réjane , Billaudel Sylviane , Courtieu André‐Louis
Published: March 2007

Between September, 1987, and April, 1989, three techniques for the detection of respiratory syncytial virus (RSV) were compared: indirect immunofluorescence (IF) on the sample, indirect immunofluorescence after 72 h of MRC‐5 cell culture (IF 72h), and detection of the cytopathic effect (CPE) by MRC‐5 and HEp‐2 cell culture. A study of 383 nasal aspirates from young children admitted to the Centre Hospitalier de Nantes (CHR) showing miscellaneous respiratory symptoms produced the following results: 143 samples (37%) were RSV positive by IF, 119 (31%) were positive by IF 72h, and 117 (31%) showed RSV‐induced CPE. In comparison with tissue culture isolation (TC), the sensitivities of IF and IF 72h were 89% and 80% and their specificities 85% and 91%, respectively. During the winter of 1988–1989, of the 110 RSV‐positive nasal aspirates (104 by IF, 89 by IF 72h, 83 by CPE detection), 109 were identified by IF and/or IF 72h. IF 72h affords rapid detection of RSV.
Acute haemorrhagic conjunctivitis: Seroepidemiology of coxsackievirus A24 variant and enterovirus 70 in Singapore
Authors: Goh K. T. , Ooi P. L. , Miyamura K. , Ogino T. , Yamazaki S.
Published: March 2007

Epidemics of acute haemorrhagic conjunctivitis caused by a variant of coxsackievirus A24 (CA24v) and enterovirus 70 (EV70) have occurred periodically in Singapore. A seroepidemiological survey conducted before the CA24v epidemic in 1985 and in the midst of the EV70 epidemic in 1983 showed a neutralising antibody prevalence (⩾ 1:4) of 19.1% and 66.9% to CA24v and EV70, respectively. The seropositivity rate to both viruses was highest in children 10–14 years of age, but no sex or ethnic difference was noted. It appears from the data that an epidemic could be triggered when the herd immunity of the population falls below a critical level. There was a significant correlation in seroprevalence between CA24v and EV70 (P = 7.75 × 10−3).