bluuebunny/crossref_metadata_2025 huggingface dataset:
Detection of human papillomavirus DNA in cell scrapes and formalin‐fixed, paraffin‐embedded tissue of the uterine cervix by filter in situ hybridisation
Authors: Demeter T. , Kulski J. K. , Rakoczy P. , Sterrett G. F. , Pixley E. C.
Published: March 2007

Filter in situ hybridisation (FISH) was used to detect the presence of DNA of human papillomavirus (HPV) types 6/11 or 16/18 in cell scrapes (CYTOFISH) and formalin‐fixed, paraffin‐embedded biopsies (HISTOFISH) taken from the uterine cervices of 19 women. Paraffin tissue sections collected for HISTOFISH were either digested with pepsin or lysed with alkali/Triton X‐100. The digest or lysate of the tissue sections and cell scrapes were applied to nylon or nitrocellulose membrances for nucleic acid hybridisation using 32P‐labeled HPV‐DNA probes. CYTOFISH and HISTOFISH were compared directly by taking samples for each method from the cervices of the same women.

Of 19 women examined by colposcopy, cytology, and histology, eight were assessed as normal and 11 had evidence of a cervical disorder and/or the presence of HPV infection. Whereas no HPV‐DNA was detected in the normal cases, the presence of HPV‐DNAs was detected by both CYTOFISH and HISTOFISH in 11 cases with histological evidence of HPV infection and/or dysplasia. In these HPV positive cases, eight contained HPV 16/18, two HPV 6/11, and one a mixed infection of HPV 6/11/16/18. The high correlation between the results of CYTOFISH and HISTOFISH shows that formalin‐fixed, paraffin‐embedded cervical biopsies are suitable specimens for the detection and typing of HPV‐DNA by FISH. Both CYTOFISH and HISTOFISH should facilitate studies on the prevalence and distribution of HPVs and their association with neoplasia.
Immunoassay for serologic diagnosis of influenza type A using recombinant DNA produced nucleoprotein antigen and monoclonal antibody to human IgG
Authors: Harmon Maurice W. , Jones Ian , Shaw Mike , Keitel Wendy , Reimer Charles B. , Halonen Pekka , Kendal Alan P.
Published: March 2007

Influenza type A nucleoprotein (NP) derived from the full length cloned gene expressed in E. coli was evaluated in a solid phase enzyme immunoassay (EIA) for detection of human antibody to influenza. Monoclonal antibody to human IgG was used for detection. Direct and indirect assays were developed and sera were tested in serial and single dilution formats. Preliminary results indicated that recombinantand virion‐derived NP antigens were comparable in binding ability to plastic and binding human antibody. Eighty‐seven paired sera from influenza patients were tested. The most sensitive assay (indirect‐serial dilution) detected 56 (64%) rises and the simplest assay (direct‐single dilution) detected 43 (49%) rises, compared to 36 (41%) for complement fixation. Paired sera from 18 control patients showed no evidence of antibody rises by any of the assays. Forty‐nine paired sera from influenza B infected patients were negative for antibody rises except for one borderline rise by the indirect‐serial dilution assay. These results indicate that the use of recombinant DNA derived nucleoprotein for immunoassay is feasible. The sensitivity of immunoassays using NP adsorbed to the solid phase and monoclonal antibody specific for human IgG to detect bound antibody exceeded that of conventional complement fixation testing for establishing serologic evidence of influenza type A infection.
Suppressor T‐cell activity in chronic hepatitis B‐virus infection: Relationship with the presence of HBV‐DNA in serum
Authors: Iraburu Margarita , Civeira María Pilar , Serrano Manuel , Morte Susana , Castilla Alberto , Prieto Jesús
Published: March 2007

Suppressor T‐cell activity and allogeneic T‐cell response to concanavalin A (ConA) were investigated in 46 patients chronically infected with hepatitis B virus (HBV). Thirty‐eight patients had chronic active hepatitis, seven of whom were superinfected with Delta virus, and eight were healthy chronic HBV carriers. T‐cell suppressor activity was in the normal range in healthy carriers and in patients negative for serum HBV‐DNA, independent of the e antigen status. In contrast, the group of patients positive for HBV‐DNA exhibited a significant reduction in suppressor activity. Longitudinal studies in patients who cleared serum HBV‐DNA demonstrated that suppressor T‐cell activity became normal thereafter. These results suggest a relationship between suppressor T‐cell function and the stage of viral replication in individuals with chronic HBV infection.
Immunogenicity of a yeast‐recombinant hepatitis B vaccine in high‐risk children
Authors: Rumi M. G. , Romeo R. , Bortolini M. , Gringeri A. , Zanett A. R. , Mannucci P. M. , Colombo M.
Published: March 2007

Horizontal transmission of hepatitis B virus (HBV) from illicit drug users to their contacts, including young children, can be prevented by active immunization against HBV. Yeast‐recombinant hepatitis B vaccines are now available for this purpose, but their potential efficacy in such high‐risk contacts has not yet been evaluated. Therefore we gave 20 mcg of a recombinant yeast‐derived hepatitis B vaccine to 38 children who were at high risk for HBV infection because they had been institutionalized in a community for drug users in which 8.7% of the occupants are carriers. After third dose of vaccine (at 0, 1, and 6 months), all children had anti‐HBs responses with titers of 10 mlU/ml or more, with 81% showing responses greater than 1,000 mlU/ml. At 12 months, the percentage of anti‐HBs‐positive children was 100%, and the percentage of children with anti‐HBs higher than 1,000 mlU/ml was 56%. None of the children developed HBV infection during follow‐up. Hence the recombinant vaccine was immunogenic, with percentages of seroconversion and anti‐HBs titers comparable with those attained in other categories of high‐risk children with plasma‐derived vaccines.
Nosocomial outbreak of infant rotavirus diarrhea due to the appearance of a new serotype 4 strain
Authors: Matteo Angela Di , Sarasini Antonella , Scotta M. Serenella , Parea Maurizio , Licardi Giovanna , Gerna Giuseppe
Published: March 2007

An outbreak of acute gastroenteritis, involving 30 infants and young children aged 2 months to 4 years, took place in a pediatric ward of the University Hospital of Pavia, Northern Italy, in the period from November 9 to December 1, 1986. Out of the 14 patients examined, ten were found to shed rotavirus with stools. All strains were characterized for serotype, using a monoclonal antibody‐based enzyme‐linked immunosorbent assay, and for electropherotype, by polyacrylamide gel electrophoresis of genomic RNA. It was shown that a single serotype 4 (subtype 4A) strain spread within the ward from a primary case to seven other patients. The remaining two patients were found to be infected by a serotype 1 strain that was circulating in the same area prior to the outbreak. The clinical symptoms were unusually severe, since significant dehydration was observed in four of the eight serotype 4 rotavirus‐infected children. Previous epidemiological studies had shown that since 1983 serotype 4 strains had not been circulating in Pavia, and the electropherotype of the newly circulating serotype 4 strain was different from those observed in 1981–1983. Thus, the severity of the diarrheal disease appeared to be related to the circulation of both a new serotype and a new electropherotype.
Titration of igg antibodies against varicella zoster virus before bone marrow transplantation is not predictive of future zoster
Authors: Webster A. , Grint P. , Brenner M. K. , Prentice H. G. , Griffiths P. D.
Published: March 2007

Serum antibodies to varicella zoster virus (VZV) were measured in 77 patients about to undergo allogeneic bone marrow transplantation, and in 65 of their donors. Ten patients developed zoster within the first 6 months following transplant. There was no significant difference in the mean pretransplant antibody titre between those patients who did or did not subsequently develop zoster. Likewise, the level of antibody to VZV amongst donors had no effect on the subsequent development of zoster. We conclude that the pretransplant level of antibody to VZV is not predictive of subsequent zoster infection, and would not be helpful in identifying patients for trials of antiviral prophylaxis. These results contrast with those previously found for another herpesvirus, herpes simplex (HSV), where antibody level pretransplant is predictive of future HSV recurrence.
Antigenic variation of human RSV strains isolated in Japan
Authors: Tsutsumi Hiroyuki , Nagai Kazushige , Suga Kazuhiro , Chiba Yasuo , Chiba Shunzo , Tsugawa Satoshi , Ogra Pearay L.
Published: March 2007

Antigenic variations of respiratory syncytial virus (RSV) strains were analyzed using a collection of nine, seven, two, and one monoclonal antibodies (MAbs), respectively, raised against the fusion protein (F), large glycoprotein (G), nucleoprotein (NP), and phosphoprotein (P) components of the Long strain of RSV. Competitive binding assay by these MAbs demonstrated eight, four, and two distinct epitopes on F, G, and NP components, respectively. Comparison of prototype Long with ten field strains isolated in Sapporo, Japan, during a 9‐year period from 1980 to 1988 by radioimmunoprecipitation (RIP), immunofluorescence (IF), and enzyme‐linked immunosorbent assay (ELISA) test revealed four different patterns of reaction to these MAbs. Thus, prototype Long reacted to all 19 MAbs. Six field strains have shown a different reactivity to one of nine anti‐F and to one of seven anti‐G antibodies (subgroup A). Three of the remaining isolates failed to react with three of nine anti‐F and with all of seven anti‐G antibodies (subgroup B). One strain (58–104) isolated in 1983 was similar to subgroup A except for a lack of reaction with two anti‐G antibodies. All field strains reacted with two anti‐NP and one anti‐P antibodies. The numbers of altered epitopes in subgroup A were 1/8 and 1/4; in subgroup B, 3/8 and 4/4; and in 58–104, 1/8 and 2/4 on the F and G components, respectively. No other variations have been observed among field isolates tested.
Serum IgA antibodies in HSV asymptomatic genital infections
Authors: Guglielmino S. P. P. , Garozzo A. , Pinizzotto M. R. , Furneri P. M. , Corbino N. , Castro A. , Cianci S.
Published: March 2007

Cervico‐vaginal swabs and serum samples from 81 women without herpes‐associated clinical symptoms were examined for virus isolation and for the presence of IgA antibodies to herpes simplex virus (HSV) using the indirect fluorescent antibody technique. Nineteen of the 81 women were followed up for 2 years. Blood samples and swabs from cutaneous lesions were obtained from another group of 20 women (medical staff) with herpes labialis. By analysing the specific anti‐HSV IgA antibodies, a higher geometric mean titre (GMT) was found in women with positive HSV‐2 isolation (GMT = 380.03), as opposed to the GMT observed in women with negative HSV‐2 (GMT = 63.43) or positive HSV‐1 isolation (GMT = 55.15). Results from the longitudinal study also demonstrated that positive virus isolation always correlated with a marked rise in serum IgA antibody to HSV.
Definition of high‐proficiency serological markers for diagnosis of Varicella‐zoster virus infections by enzyme immunoassay
Authors: Echevarría José M. , de Ory Fernando , León Pilar , Téllez Alicia
Published: March 2007

The levels of Varicella‐zoster virus (VZV)‐specific IgG, IgM, and IgA antibodies produced in 22 cases of varicella, 22 cases of cutaneous zoster, and 12 cases of acute aseptic meningitis due to VZV in the absence of cutaneous lesions, were measured by indirect enzyme immunoassay (EIA) and compared with those observed in a group of 34 age‐matched controls. The definition of cutoff titres for each serological marker and combinations of them allowed early diagnosis of infection in 82% of varicella patients and 91% of patients with acute aseptic meningitis lacking cutaneous lesions on a single serum sample, the specificity being over 90%. The system was as sensitive as the demonstration of intrathecally produced IgG antibodies for the early diagnosis of the infection in 22 cases of neurological disease due to VZV. A working protocol for the serological diagnosis of VZV infections, using currently available EIA reagents, is described.
Heterophile igm, iga, and ige antibodies in infectious mononucleosis
Authors: Duverlie G. , Driencourt M. , Roussel C. , Orfila J.
Published: March 2007

Fifty infectious mononucleosis (IM) and 150 noninfectious mononucleosis (non‐IM) sera were tested by a hemadsorption immunocapture test (HIT) for the detection of heterophile antibodies of IgM, IgA, and IgE classes. The specificity of Paul‐Bunnell (PB) antibodies was ascertained by a differential absorption test. IgM PB‐antibodies were demonstrated in 100% of IM sera, IgA in 92%, and IgE in 88%. PB antibodies were present only in IM sera; but other heterophile antibodies of IgM (68%), IgA (6.7%), and IgE (9.3%) classes were demonstrated in non‐IM sera. IgA and IgE heterophile antibodies were thus present, especially in IM sera. HIT was found to be more sensitive than Paul‐Bunnell Davidsohn test (PBD test) and suitable for the diagnosis of infectious mononucleosis.
Amplification of rhinovirus specific nucleic acids from clinical samples using the polymerase chain reaction
Authors: Gama R. E. , Horsnell P. R. , Hughes P. J. , North C. , Stanway G. , Bruce C. B. , Al‐Nakib W.
Published: March 2007

We describe a novel method for the detection of human rhinoviruses in clinical samples, using the polymerase chain reaction. Two synthetic oligonucleotide primers were produced that bind in the 5′ noncoding region of all rhinovirus serotypes tested, about 350 nucleotides apart, and were used to prime polymerase chain reaction amplification of the intervening stretch of DNA. The product of this reaction, which can be clearly visualized by gel electrophoresis, is a discrete 380 bp band, the occurrence of which is diagnostic of the presence of a rhinovirus in the clinical sample analysed.

The technique, which is rapid, sensitive, and reliable, has been used successfully for all the different rhinovirus serotypes tested to date in our laboratory. However, the sensitivity of detection is greatly dependent on the inclusion of both tRNA and vanadyl complexes during the viral RNA extraction process. Using this technique, under optimal conditions, we were able to detect virus in clinical samples with titres as low as TCID50 102.5.
Shedding of herpes simplex virus type 1 into saliva in patients with orofacial fracture
Authors: Kameyama T. , Futami M. , Nakayoshi N. , Sujaku C. , Yamamoto S.
Published: March 2007

Shedding of herpes simplex virus type 1 (HSV‐1) into saliva was studied in 83 patients with orofacial fractures. Infectious virus was isolated from 14 of 83 patients (16.8%) with no detectable herpetic lesion during hospitalization. Of the 83 patients, 44 (53%) had HSV‐1 specific antibody. Virus shedding into saliva was observed only in the patients with antibody to HSV‐1; thus, the frequency of virus shedding patients among those with antibody was 31% (14 of 44 patients).

The frequency was obviously higher than that of a healthy population. The period of HSV‐1 shedding had a mean of 3.7 days with range of 1 to 8 days, which is also significantly longer than that of a healthy population. These results strongly suggest that both treatment and operation lead effectively to reactivation of latently infected HSV‐1.
Viral release from hiv‐i‐induced syncytia of cd4+ c8166 cells
Authors: Harris J. R. , Kitchen A. D. , Harrison J. F. , Tovey G.
Published: March 2007

Infection of the permissive C8166 CD4+ lymphoid cell line with human immunodeficiency virus (HIV)‐I rapidly leads to syncytium formation. Ultrastructural features of medium‐sized (40 to 60m̈m) syncytia and larger syncytia are presented, with emphasis upon HIV‐I release from the syncytial surface and into intrasyncytial vacuoles. Although surface release of HIV‐I is shown to diminish with increasing syncytial size, it does not totally stop. Massive HIV‐I release and entrapment within intrasyncytial vacuoles is clearly apparent within the “foamy” multivacuolated zones within medium and large syncytia. Numerous multicored aberrant viruses are routinely detected within the intrasyncytial vacuoles, and double‐budding events likewise are readily detectable at the vacuolar membranes, but less so at the syncytial surface. It is suggested that this observation may be a reflection of the more restricted availability of vacuolar membrane area for excessive viral budding as compared with that at the syncytial surface.
Antibody against epstein‐barr virus dna polymerase activity in sera of patients with nasopharyngeal carcinoma
Authors: Liu Mei‐Ying , Chou Wen‐Hsiang , Chen Jen‐Yang , Yang Czau‐Siung , Nutter Louise , Hsu Mow‐Ming
Published: March 2007

A salt‐dependent DNA polymerase activity was demonstrated in the culture of an EBV‐producing, lymphoblastoid cell line (NPC‐204 cells) treated with 5‐iodo‐2′‐deoxyuridine (IUdR). There was a high frequency of levels of antibody to this enzyme in sera of patients with nasopharyngeal carcinoma (NPC). In contrast, sera from healthy subjects had little or no neutralizing activity. The high antibody level appeared as early as stage 1 of the disease in many NPC patients. The levels of the antibody increased with the progression of the disease and declined in treated patients. The results strongly suggest that tests measuring serum antibody against EBV DNA polymerase activity can be used for early diagnosis and prognosis of NPC.
Characterization of cytomegalovirus isolates recovered during repeated infection in renal transplant recipients
Authors: Kanesaki Takumi , Baba Koichi , Tanaka Kazuko , Ishibashi Michio , Yabuuchi Hyakuji
Published: March 2007

The sources of cytomegalovirus (CMV) infection in seropositive renal transplant recipients include reactivation of latent endogenous virus in the recipient, reactivation of latent virus in donated kidney, or both. If infection occurs by either source, viruses isolated from the same recipient should be the same strain, whereas if it occurs by both sources, those from the same recipient should be different. In this study, we followed prospectively 25 seropositive recipients who predominantly received a kidney from a seropositive donor to determine whether CMV isolates recovered repeatedly from them are the same or different. During an average 10 month follow‐up period, six (24%) patients had excreted the virus more than twice at any site and/or at different times. Restriction enzyme analysis of viral DNA prepared by the Hirt procedure revealed that three or four isolates obtained from each of five patients were same, whereas six isolates from one patient included three different strains. Five of six patients had clinical symptoms at the times when CMV was recovered. Three patients with acute rejection and one patient with hepatitis had been infected with one single strain, and all were successfully treated. One patient with fever and acute rejection had been infected with three different strains, and he failed to recover his renal function. These results suggest that most CMV infections in seropositive recipients may be caused by one single strain. However, multiple infections with different strains can also occur. Such infections are associated with more severe clinical disease.
Dissociated antibody responses to the s and pre‐s2 regions of the hepatitis b virus after vaccination in hemophiliacs
Authors: Zanetti Alessandro , Tanzi Elisabetta , Mannucci Pier Mannuccio
Published: March 2007

The membranes of hepatocytes and the pre‐S2 envelope protein of the hepatitis B virus (HBV) contain binding sites for polymerized human albumin, which is thought to act as a link between HBV and hepatocytes. Hence, anti‐pre‐S2 antibodies should prevent HBV uptake by the liver, and there is indeed preliminary evidence that they protect chimpanzees from HBV infection. To evaluate whether a plasma‐derived vaccine containing the pre‐S2 sequence induced an antipre‐S2 response in 105 vaccinated hemophiliacs, anti‐pre‐S2 was measured in parallel with antibody to hepatitis B surface antigen (anti‐HBs). Eighty‐five percent of the hemophiliacs had both anti‐pre‐S2 and anti‐HBs when vaccination was completed, 13% had anti‐HBs alone, and 2% (two cases) had anti‐pre‐S2 alone. Eighty‐seven percent of anti‐pre‐S2‐positive hemophiliacs compared with only 50% of anti‐pre‐S2‐negative hemophiliacs (P< 0.001) developed high anti‐HBs titers (≥ 1,000 mlU/ml). This study demonstrates, therefore, that the antibody responses to the S and pre‐S2 regions of HBV may be dissociated after vaccination in hemophiliacs and that higher anti‐HBs titers are attained in anti‐pre‐S2‐positive hemophiliacs.
Clearance of hepatitis b virus dna and pre‐s surface antigens in patients with markers of acute viral replication
Authors: Delfini C. , Colloca S. , Stroffolini T. , Taliani G. , Mazzotta F. , D'Agata A. , Buonamici C. , Carloni G.
Published: March 2007

To clarify the relationship between the pre‐S antigens and other serological markers of hepatitis B virus (HBV) replication, we followed up 27 patients: 21 presented with symptoms of acute hepatitis (two progressed to chronicity) and six suffered from chronic hepatitis. Pre‐S1, pre‐S2, HBV DNA, IgM antihepatitis core antigen (HBc), hepatitis B e antigen(HBeAg), and anti‐HBe were detected in about 200 sera serially collected at different times for at least 6–12 months from the onset of clinical observation. In the early symptomatic phase of acute hepatitis, the pre‐S1and pre‐S2antigens were present in 95% of the cases and correlated well with high levels of alaninetransferase (ALT) and IgM anti‐HBc, while HBV DNA was present in the sera of only six (28.6%) patients (P< 0.0001). This was the first marker to disappear (1 month after the initial stage). All of the HBV DNA‐positive patients were also HBeAg positive, whereas no HBeAg‐negative subjects were found with serum HBV DNA. In the six chronic patients, pre‐S antigens were always present independently of the HBeAg/anti‐HBe status; HBV DNA was detected in three of them, even if transiently, and in two of these it reappeared together with the pre‐S2epitope. The follow‐up data suggest that, in acute hepatitis, the clearance of pre‐S antigens can be considered as a prognostic index of clinical resolution and that, in chronic hepatitis, the persistence of pre‐S antigens seems to indicate progression of the disease. In particular, pre‐S2, in patients in whom it is intermittent, can be considered as an index of reactivation.
Predilection of a nasopharyngeal carcinoma‐derived isolate of epstein‐barr virus for infection of specific subsets of B lymphocytes
Authors: Tarr Kathleen L. , Glaser Ronald
Published: March 2007

It is important to know whether there are variants of Epstein‐Barr virus (EBV) with biological properties that are different from the prototype viruses that have been studied in detail, such as P3HR‐1 and B95‐8. We have studied an EBV isolate derived from a nasopharyngeal carcinoma (NPC) tumor, designated NPC‐EBV. We have examined the target B lymphocytes infected and growth‐transformed with NPC‐EBV as compared with two common EBV isolates, B95‐8 and AG876 EBV, for stage of maturation using antibodies to several immunoglobulin chains. Typing of the NPC‐EBV transformed lymphoblastoid cell lines revealed the predilection of the NPC‐EBV isolate to infect immature B lymphocytes. This was not the case for the B95‐8 and AG876 isolates. The reason for the predilection of NPC‐EBV for immature B lymphocytes remains to be explored further. However, these results may be important in understanding the pathophysiology of EBV‐associated diseases.
Elimination of circulatory IGM anti‐HSA precedes anti‐HBe seroconversion in patients with CAH type B
Authors: Hellström U. B. , Sylvan S. P. E.
Published: March 2007

The presence of IgM and IgA antibodies with specificity for human serum albumin (HSA) were consecutively analyzed in serum samples from four patients with biopsy verified CAH type B during seroconversion in the HBe/anti‐HBe antigen system. Circulatory IgM anti‐HSA antibodies were present during HBe antigenemia. The antibody titers fluctuated, decreased, and were finally lost from the circulation. After the disappearance of IgM anti‐HSA antibodies, seroconversion to anti‐HBe reactivity occurred and a quiescent phase of the disease was established, as judged by normalization of transaminases and absence of circulatory HBV‐DNA.

IgA anti‐HSA antibodies persisted in the circulation after the elimination of IgM anti‐HSA and seroconversion to anti‐HBe reactivity. For one of the patients, a dramatic increase in titers was followed by elimination of IgA anti‐HSA and seroconversion to anti‐HBs.

The data indicate that the host “self”‐component HSA, when associated with “foreign” HBe or HBs antigenic structures, elicit immune responses to HSA, preventing the adequate development of anti‐HBe and anti‐HBs. The cessation of anti‐HSA reactivity, however, seemed to permit subsequent sensitization to HBe and HBs antigenic determinants, as detected by the presence of circulatory antibodies.
Active replication of human immunodeficiency virus type 1 by peripheral blood mononuclear cells following coincubation with herpes viruses
Authors: Tremblay Michel , Gornitsky Mervyn , Wainberg Mark A.
Published: March 2007

Patients with acquired immunodeficiency syndrome (AIDS) commonly suffer from opportunistic infections associated with members of the herpes virus family. To investigate whether certain of these other viruses might have an effect on the ability of the human immunodeficiency virus type 1 (HIV‐1) to replicate, we coincubated peripheral blood mononuclear cells (PBMC) from nine HIV‐1‐seropositive donors with live preparations of various herpes viruses. In seven of nine cases, exposure of PBMC to preparations of either HSV‐1, HSV‐2, or CMV stimulated the cells to become active producers of HIV‐1, as determined by reverse transcriptase activity and by the presence of infectious progeny virus. This increased production of HIV‐1 particles appeared to be a consequence of mitogenic proliferation and of herpes virus‐encoded transacting factors. These results supplement earlier findings on the molecular activation of the HIV‐1 genome by both HSV and CMV genetic elements and point to a possible role for these viruses in the pathogenesis and ultimate clinical outcome of HIV‐1 infections.
Coronavirus‐like particles in adults in melbourne, Australia
Authors: Marshall J. A. , Thompson W. L. , Gust I. D.
Published: March 2007

Coronavirus‐like particle(s) (CVLP) are faecal‐derived pleomorphic membrane bound virus‐like particles characterised by a fringe of clubshaped spikes that measure about 27 nm in length. The association of CVLP with a variety of social, clinical, and epidemiological factors was examined after a 69 month survey of faeces received for routine testing at an infectious diseases hospital. CVLP was found most commonly in three groups: first, intellectually retarded individuals who were usually inmates of institutions; second, recent overseas travellers who were either Indochinese refugees/immigrants or were overseas travellers who had usually visited developing communities for lengthy periods; and, third, male homosexuals who had a history of multiple sexual contacts and/or venereal disease. It was concluded that the excretion of CVLP had a strong association with unhygienic living or working conditions irrespective of any clinical symptoms the individual might show.
Effect of 3′‐azido‐3′‐deoxythymidine on replication of duck hepatitis B virus in vivo and in vitro
Authors: Haritani Hideaki , Uchida Toshikazu , Okuda Yasunori , Shikata Toshio
Published: March 2007

3′‐Azido‐3′‐deoxythymidine (AZT) inhibits the replication of the human immunodeficiency virus (HIV) by blocking the formation of the phosphodiester bond and has been used clinically for the treatment of HIV infection. To assess the effect of AZT on the replication of hepadnaviruses, which replicate through reverse transcription, both the liver tissue and primary cultured hepatocytes from ducklings previously infected with duck hepatitis B virus (DHBV) were examined for DHBV DNA before and after the treatment with AZT. We did not observe suppression of DHBV replication at any doses in our system as measured by viral DNA synthesis in infected duck hepatocytes. The data strongly suggest that AZT has no inhibitory effect on DHBV reverse transcriptase.
Immune blot analysis of viral surface proteins in serum and liver of patients with chronic hepatitis B virus infection
Authors: Gerken Guido , Manns Michael , Gerlich Wolfram H. , Hess Georg , Zum Büschenfelde Karl‐Hermann Meyer
Published: March 2007

The small and the middle surface proteins of hepatitis virus form either the virion or the 22 nm particle both of which are secreted. The large surface protein by itself remains cell bound in artificially transfected cell culture unless it is accompanied by an excess of the smaller proteins. Its behavior in vivo is not yet well studied. Using specific monoclonal antibodies for immunoblotting, we found an abundance of small surface protein in the serum of chronic virus carriers and moderate amounts in the liver irrespective of viremia. The large surface protein was present in the serum and the liver of viremic carriers. In nonviremic carriers, the large protein was absent from serum, but in the liver a shorter form of the large protein was readily detectable. These findings suggest a complex regulatory mechanism of the viral surface protein depending on the expression of other viral gene products.
Epidemiology of human T‐Cell leukemia/lymphoma virus type 1 (HTLV‐1) infections in a subpopulation of Afro‐Caribbean origin in England
Authors: Mowbray Janet , Mawson Sharron , Chawira Atha , Skidmore Susan , Boxall Elizabeth , Desselberger Ulrich , Nightingale Simon
Published: March 2007

Epidemiological studies on neurological diseases in residents of Afro‐Caribbean origin in the West Midlands region of England have identified eight patients with tropical spastic paraparesis (TSP), all of whom were found to be infected with human T‐cell leukemia/lymphoma virus type 1 (HTLV‐1). The husband of one of the patients with TSP was also infected with HTLV‐1 and had a T‐cell lymphoma. In addition, six asymptomatic HTLV‐1‐infected first‐degree relatives of the TSP patients have been found. By anonymous testing of over 700 sera obtained from individuals of Afro‐Caribbean, African, or Asian ethnic origin, seven HTLV‐1‐infected individuals were detected, who were all immigrants from the Caribbean. Overall, these numbers yielded a seroprevalence of HTLV‐1 infections of 3.4% among the immigrant population of Afro‐Caribbean origin, which is comparable with the prevalence of HTLV‐1 in Jamaica in an equivalent age and sex cohort. Sera were tested for HTLV‐1 antibody by means of three different procedures: passive particle agglutination test (Serodia), indirect enzyme‐labeled immunosorbent assay (ELISA; Dupont), and indirect immunofluorescence test (in‐house, using HTLV‐1‐infected MT2 cells). The results of all three tests correlated very well with each other. HTLV‐1 antibody titres in TSP patients were on the whole significantly higher than those of asymptomatic carriers, but some of the apparently healthy firstdegree relatives and one anonymously tested individual had titres as high as most of the TSP patients.
An outbreak of acute flaccid paralysis in jamaica associated with echovirus type 22
Authors: Figueroa J. P. , Ashley D. , King D. , Hull B.
Published: March 2007

An outbreak of acute flaccid paralysis in Jamaica in 1986 associated with echovirus type 22 is described. Six patients aged 1 to 27 years developed acute onset of severe flaccid paralysis, with inability to walk. Three cases had facial weakness, four required intensive care with assisted ventilation, and two died. Echovirus type 22 was isolated from the stool of two patients who showed a significant increase in antibody titre. Echovirus type 22 was also isolated from the stool of another patient who had aseptic meningitis without any neurological deficit. There was no evidence of poliovirus infection in any of these patients, most of whom were fully immunized. Of the four surviving cases with flaccid paralysis, three had residual weakness in their lower limbs and walked with an abnormal gait 3 years after the acute paralytic attack. This is the first report in the literature of acute flaccid paralysis associated with type 22 echovirus.
Comparison of class and subclass distribution of antibodies to the hepatitis B core and B e antigens in chronic hepatitis B
Authors: Sällberg Matti , Norder Helene , Magnius Lars O.
Published: March 2007

The IgG subclasses IgM and IgA1 of antibodies to hepatitis B core antigen (anti‐HBc) and hepatitis B e antigen (anti‐HBe) were assayed in sera from 82 patients with chronic hepatitis B utilising class/subclass‐specific enzyme immunoassays (EIA). The solid‐phase was either recombinant hepatitis B core antigen (rHBcAg) or rHBcAg converted to HBeAg by addition of 0.1% SDS with remaining HBcAg antigenicity blocked with monoclonal anti‐HBc. Anti‐HBc IgG1 was detected in 81 sera at a geometrical mean titre (GMT) of 296, 110 2.9. Anti‐HBc IgG2 was not detected in any of the sera, and anti‐HBc IgG3 and IgG4 were detected in 50 and 37 sera, respectively. Anti‐HBc IgM and IgA1 were both significantly correlated to the presence of HBV DNA. The predominant antibody to HBeAg was found to be IgG1, being detected in 45 sera with a GMT of 1,035 3.3. Anti‐HBe IgG2 was not detected in any serum, while anti‐HBe IgG3 and IgG4 were found in 8 and 23 sera, respectively. Anti‐HBe IgG1, IgG3, and IgG4 were mainly detected in sera positive for anti‐HBe in RIA (Abbott). No patient was found positive for anti‐HBe IgA1 or IgM. Thus, in contrast to HBcAg, HBeAg does not trigger a persistent IgM and IgA1 response in chronic hepatitis B. The levels of anti‐HBe IgG1 and IgG3 were much lower than the levels of anti‐HBc IgG1 and IgG3. The presence of anti‐HBe IgG4 was significantly correlated to that of anti‐HBc IgG4. The relative levels of anti‐HBc IgG subclasses were found to be IgG1 > IgG3 > IgG4, whereas the relative levels of anti‐HBe subclasses were IgG1 ≥ IgG4 ≥ IgG3. The anti‐HBe IgG1:IgG4 ratio was much lower than that of anti‐HBc, 2.7:1 versus 890:1. Our findings support the suggestion of differences at the B‐cell level in the regulation of the immunoresponse to HBeAg and HBcAg, though both antigenic epitopes reside on the same polypeptide.
Expression of pre‐S2 region of hepatitis B surface antigen inEscherichia coli
Authors: Yu M. W. , Shih J. W.‐K. , Bursztyn‐Pettegrew H. , Byars N. E. , Allison A. C. , Chan H. W.
Published: March 2007

We constructed a recombinant plasmid that can express the entire pre‐S2 sequence of hepatitis B surface antigen (HBsAg) as a fusion protein inE. coli. The hybrid protein, which comprises the bacterial TrpLE protein and the pre‐S2 sequence, was the prominent protein that was found in cell extracts. As determined by immune blot analysis, this protein reacted with human HBV convalescent sera, as well as with sera from animals immunized with either purified HBsAg or isolated polypeptides containing pre‐S2. It bound specifically to125I‐polymerized human albumin cross‐linked with glutaraldehyde but not to125I‐monomeric human albumin. A novel adjuvant formulation was used in place of Freund's adjuvant to immunize guinea pigs with the recombinant product. The antisera obtained from serial bleedings were found to react with HBsAg of both d and y subtypes. These antisera were also shown to react solely with HBsAg polypeptides which contain the pre‐S2 sequence and to block the binding of HBsAg to solid‐phase polymerized human albumin.
Neutralizing antibody assay for human Herpesvirus‐6
Authors: Suga Sadao , Yoshikawa Tetsushi , Asano Yoshizo , Yazaki Takehiko , Ozaki Takao
Published: March 2007

Antibody to human herpesvirus‐6 (HHV‐6) was measured in cord blood mononuclear cell cultures by a neutralization (NT) test, in which the presence or absence of characteristic large cell formation in cells infected with HHV‐6 was used as an indicator for neutralization of the virus. The NT test could measure antibodies during and just after the appearance of the skin rash in patients with exanthem subitum. The levels of antibodies measured by the NT test was generally higher than that by an indirect immunofluorescence assay.
Immunoglobulin and hepatitis B surface antigen‐specific circulating immune complexes in chronic hepatitis with hepatitis delta virus infection
Authors: Tsai Jung‐Fa , Margolis Harold S. , Fields Howard A. , Nainan Omana V. , Chang Wen Yu , Tsai Juei‐Hsiung
Published: March 2007

IgM, IgG, and HBsAg containing circulating immune complexes (CIC) were determined, by conglutinin (K) and C1q assays, for assessing the role of CIC in hepatitis delta virus (HDV) infection in 54 HBsAg‐negative controls and 85 HBsAg‐positive patients with chronic hepatitis. The prevalence of HDV markers (HDV antigen and anti‐HD) was 24.70% (21/85). CIC were a common feature of HDV infection with 95.24% of patients having at least one abnormal test resutlt. The prevalence of elevated IgM‐K, IgG‐K, IgM‐C1q, and IgG‐C1q CIC were 85.71, 85.71, 57.14, and 85.71%, respectively. The prevalence of IgM class CIC were statistically higher in patients with HDV infection than in those without (P= .001 for the K assay andP= .023 for the C1q assay). There was no difference in the prevalence of IgG class CIC. Patients with HDV infection also have significantly higher median levels of IgM K‐CIC (P= .002), IgG K‐CIC (P= .049), and IgG C1q‐CIC (P= .008). In patients with HDV infection, there was positive correlation between IgM C1q‐CIC and transminase levels (r= .519,P= .016 for AST;r= .500,P= .021 for ALT). There was no difference in the prevalence of HBsAg containing CIC between patients with HDV infection (76.19%) and those without (74.60%). In conclusion, IgM class CIC are the major CIC and correlate with disease activity in HDV infection. CIC may play a role in the pathogenesis of HDV infection.
Serial passage of West‐European sporadic non‐A non‐B hepatitis in rhesus monkeys by inoculation with fecal extracts
Authors: Sharma Madhav D. , Maillard Patrick , Vrati Sudhanshu , Mukherjee Rama , Rajagopalan M. S. , Poynard Thierry , Talwar Gurusaran P. , Pillot Jacques
Published: March 2007

An experimental model of sporadic non‐A non‐B hepatitis involving a Fab nonimmune binding activity in stools was established in the rhesus monkey. The first animal was inoculated intravenously with a stool extract from a French patient who had never left the country and in whom post‐transfusion hepatitis was excluded. Four passages were performed, and the infection was transmitted by parenteral as well as the oral routes by inoculation of stools or liver extracts. Infection led in three monkeys to reversible hepatocyte injury manifested by a transitory increase in serum aminotransferases. The other three animals, in which persistently high levels of aminotransferases was observed, were sacrificed on day 60 after inoculation. The incubation period, as evidenced by elevation of aminotransferases was about 3 to 4 weeks. The infectious agent was transitorily present in the stools before aminotransferase elevation. The presence of the infectious agent in the stools was correlated with the nonimmune Fab binding activity.
A new genome type of adenovirus 3 associated with severe lower acute respiratory infection in children
Authors: Kajon Adriana E. , Murtagh Patricia , Franco Susana Garcia , Freire Maria Cecilia , Weissenbacher Mercedes C. , Zorzópulos Jorge
Published: March 2007

Adenovirus 3 was found associated with ten cases of infantile lower acute respiratory infection in patients aged 2–18 months. Of these, five had a fatal outcome, with severe lung damage. Restriction enzyme analysis with Bam HI, Bcl I, BgI II, Bst E II, Hind III, Sal I, Sma I, Xba I, and Xho I revealed the presence of the same genome type in all ten specimens. The genomic variant was different from those previously reported for serotype 3 and therefore was tentatively denominated 3f.
Hepatitis B vaccine in the prevention of perinatally transmitted hepatitis B virus infection: Final report on a West Midlands pilot study
Authors: Wheeley Susan M. , Boxall Elizabeth H. , Tarlow Michael J. , Rashid Gatrad A. , Anderson Janet , Bissenden Jeffrey , Chin Kwei Cheong , Mayne Andrea
Published: March 2007

A four‐dose vaccination schedule was used to interrupt perinatal transmission of hepatitis B virus from carrier mothers to their babies. Of 49 babies immunised and successfully followed up, 43 (88%) became immune: 15 out of 21 (71%) of babies born to HBeAg+ mothers became immune, the other 6 becoming the only carrier babies in the study. Without immunisation a carrier rate in excess of 70% would have been expected in this high‐risk group. Vaccine alone, given in a rapid immunisation schedule, protected the majority of babies at risk. In those babies in whom the carrier state occurred in spite of immunisation, infection may have taken place in utero, or the infant may have failed to produce adequate antibody in response to the vaccine.
Glycoprotein gB of herpes simplex virus expresses type‐common and type‐specific antigenic determinants in vivo
Authors: Dix Richard D.
Published: March 2007

Four monoclonal antibodies directed against glycoprotein B of herpes simplex virus were evaluated for their ability to immunize mice passively against acute virus‐induced neurological illness and death when administered intraperitoneally 2 hours prior to footpad challenge with type 1 or type 2 virus. Two monoclonal antibodies, H120 and H157, failed to reduce the severity of neurological disease in infected animals. In contrast, H233 and H368 antibodies provided significant protection in type‐common and type‐specific fashions, respectively. A direct correlation was observed between in vitro neutralization and in vivo protection. These results provide the first in vivo evidence that glycoprotein gB of herpes simplex virus expresses both type‐common and type‐specific determinants during the evolution of acute virus‐induced neurological disease.
Group C rotavirus associated with fatal enteritis in a family outbreak
Authors: Caul E. O. , Ashley C. R. , Darville J. M. , Bridger J. C.
Published: March 2007

A family outbreak of gastroenteritis involving three adults and three children is described in which diarrhoea and vomiting were the main clinical features. One infant died in whom no pathogens could be detected in either small or large intestinal postmortem samples. Stool samples from two symptomatic siblings contained rotaviruses as demonstrated by electron microscopy. Both of these faecal samples were negative when assayed in a group A specific rotavirus enzyme‐linked immunosorbent assay (ELISA) and subsequent genomic analysis of these rotaviruses was suggestive of group C rotavirus. Serological evidence showed that these atypical rotaviruses were members of serogroup C. Other atypical rotaviruses in faecal samples from sporadic cases in symptomatic children were detected over a similar time period and location. These had electrophoretic RNA profiles similar to those in the family outbreak. Furthermore, seroepidemiological studies detected group C rotavirus antibody in blood donors resident in the location of the family outbreak.
Prospective study of diarrheal diseases in Venezuelan children to evaluate the efficacy of rhesus rotavirus vaccine
Authors: Perez‐Schael Irene , Garcia Dorys , Gonzalez Marino , Gonzalez Rosabel , Daoud Naimeh , Perez Mireya , Cunto Walter , Kapikian Albert Z. , Flores Jorge
Published: March 2007

The efficacy of a rhesus rotavirus vaccine (MMU 18006, serotype 3) against infantile diarrhea was evaluated by active home surveillance of a group of 320 children 1–10 months of age in Caracas, Venezuela. During a 1 year period following oral administration of vaccine or placebo under a double‐masked code, over 600 diarrheal episodes were detected. Etiologic studies revealed that heat‐stable toxin (ST) producing enterotoxigenic E. coli (ETEC) was the most common diarrheal agent detected (34%) followed by enteropathogenic E. coli (EPEC, 10.9%), heat‐labile toxin (LT) producing ETEC (7.6%), rotavirus (6.9%), Cryptosporidium (4.8%) and Campylobacter (1.3%). ST‐producing ETEC were also recovered from over 20% of control stool specimens obtained during diarrhea‐free periods, whereas EPEC, rotavirus, Cryptosporidium, and Campylobacter were rarely detected in such control specimens. Rotavirus was responsible for about one‐half of the more severe cases of diarrhea. Twenty‐two of 151 infants who received placebo (14.6%) and eight of 151 receiving a 104 PFU dose of vaccine (5.3%) had rotavirus diarrhea during the follow‐up period for an efficacy level of 64% against any rotavirus diarrhea. However, vaccine efficacy reached 90% against the more severe cases of rotavirus diarrhea and was notably high in the 1–4 month age group. Serotypic analysis of the rotaviruses detected suggests that the resistance induced by the vaccine was type specific since significant protection was only evident against serotype 3 rotaviruses. A 103 PFU dose tested initially in 18 children did not appear to protect against rotavirus diarrhea.
Postexposure treatment of experimental DHBV infection: A new therapeutic strategy
Authors: Freiman John S. , Murray Scott M. , Vickery Karen , Lim Diana , Cossart Yvonne E.
Published: March 2007

The therapeutic efficacy of antiviral agents for postexposure prophylaxis to hepadnavirus infection has been studied using acyclovir and foscarnet in the duck hepatitis B virus (DHBV) model. A total of 112 Pekin‐Aylesbury ducks were inoculated with DHBV at 11 days posthatch. Three days later, groups of these birds were injected intraperitoneally twice daily for 10 days with acyclovir (25 mg/kg) or foscarnet (250 mg/kg) or phosphate‐buffered saline. Serum samples were taken before, during, and up to 4 weeks post‐treatment and were analysed for DHBV DNA by dot hybridization. Liver tissue obtained at sacrifice was examined for viral DNA and for histological changes.

At completion of treatment with acyclovir, 21 of 22 ducks were not viremic, compared with 6 of 26 control birds (P<0.001). Four weeks after withdrawal of acyclovir, 12 of 20 ducks remained nonviremic, compared with 2 of 23 controls (P<0.01). In liver tissue, viral DNA was detected in 10 of 19 treated ducks, compared with 21/24 controls (P<0.01). Histological changes of hepatitis were present in more of the control birds than in the treated group. The results with foscarnet treatment were similar, although a smaller inoculum of DHBV was used and fewer control birds became infected.

The administration of antiviral agents soon after exposure prevented productive infection in approximately 50% of birds. Therefore, the use of a safe antiviral agent such as acyclovir, which can be given orally, should be considered in post‐exposure prophylaxis against human hepatitis B virus (HBV) infection.