bluuebunny/crossref_metadata_2025 huggingface dataset:
Shedding of coronavirus‐like particles by children in lesotho
Authors: Kidd A. H. , Esrey S. A. , Ujfalusi Maria J.
Published: March 2007

Stools from 266 children in four districts of Lesotho were examined for viruses by electron microscopy (EM) over a 5‐week period. Eighty one (30.5%) of the children shed coronavirus‐like particles (CVLPs). Shedding was not significantly associated with diarrhea. The proportion of children shedding these particles increased with increasing age. In one district, the prevalence of CVLP shedding (67.9%) was at least twice the prevalence in any of the other three districts. This was the only district to be sampled during the first week of the study. It was shown that the proportion of children shedding CVLPs declined during each of the 5 weeks of study. The stools of six children who shed CVLPs in the early summer of 1984–85 were negative by EM 5 months later. There was no association between the shedding of pathogenic bacteria or parasites and the presence or absence of CVLPs in the stool.
Calomys callidus as a potential junin virus reservoir
Authors: Videla Cristina , Kajon Adriana , Carballal Guadalupe , Weissenbacher Mercedes
Published: March 2007

The present study investigated whether C. callidus, a species belonging to the Calomys genus, is capable of developing experimentally a persistent Junin virus (JV) infection. Newborn and adult cricetids were inoculated with the attenuated XJ‐Clone 3 strain of JV by intracerebral or mucosal route. The present results indicate that the species is susceptible to JV infection, capable of shedding virus chronically through saliva and developing a persistent infection as shown by the detection of virus in brain tissue at 60 days post infection. These findings, and the fact that this cricetid shares its distribution areas with Calomys musculinus and Akodon azarae, support C. callidus as a potential JV reservoir.
Human papillomavirus coinfections of the vulva and uterine cervix
Authors: Kulski J. K. , Demeter T. , Rakoczy P. , Sterrett G. F. , Pixley E. G.
Published: March 2007

DNA filter in situ hybridisation (FISH) was used to determine the presence of human papillomavirus (HPV) genotypes 6/11, 16/18, and 31/33 in cell scrapes of the cervix and vulva of 128 women who had precancerous lesions and/or HPV infection of the cervix diagnosed by cytology, colposcopy, and histology. HPV‐DNA was found in 87 (68%) vulval and 95 (74%) cervical cell scrapes, and in both the vulval and cervical scrapes of 73 (57%) women, but not in either the vulva or the cervix of 19 women (15%). Of the HPV‐DNA‐positive smears, the prevalence of the HPV types was 61% HPV 16/18, 14% HPV 6/11, 3% HPV 31/33, and 22% HPV 6/11 and 16/18. By contrast, HPV‐DNA was not detected in the cervical smears of a control group of 35 women who were assessed to be free of cervical abnormalities by colposcopy and cytology.

The epithelial response of the vulva and the cervix to application of 5% acetic acid was assessed by colposcopy and the results correlated with the presence of HPV genotypes. A possible or definite disorder of the cervix and vulva was detected by colposcopy in 95 (74%) and 96 (75%) of the 128 cases, respectively. The colposcopic assessment of the vulva was inconclusive in ten cases (8%), and only eight women (6%) were found to be free of both a vulval and cervical disorder.

This study shows subclinical papillomavirus infections of the vulva frequently coexist with HPV infections and precancerous lesions of the cervix. Careful colposcopic examination and greater attention to HPV infections of the vulva may improve the treatment and management of precancerous lesions and HPV infections of the cervix, and help to reduce their recurrence. Should new methods of treatment of HPV disorders be developed, vulval infections will need to be treated coincidentally with those of the cervix.
Prevalence of hepatitis B markers among hospital workers in Senegal
Authors: Romieu I. , Sow I. , Lu S. , Laroque G. , Prince‐David M. , Romet‐Lemonne J. L.
Published: March 2007

A total of 775 serum samples from men and women working in hospitals in Dakar, Senegal, were tested for serological markers of hepatitis B virus (HBV). HBsAg was detected in 17.8% of the subjects, and 79.2% of the subjects were anti‐HBc positive. Among HBsAg carriers 0.04% (5) subjects were HBeAg positive, 0.03% (4) were HBV‐DNA positive, and 5.8% (8) were also anti‐Delta positive. HBsAg seropositivity was independent of sex and inversely related to age. Duration of service in the hospital was an important predictor of HBsAg seropositivity and the prevalence of seropositive subjects peaked between 2 and 3 years of employment (OR = 1.9; 95% confidence interval [CI] = 1.1–3.3, when compared to subjects who worked 1 year of less). This peak was critical in the Department of Dentistry, where subjects who worked for 2–3 years experienced a fourfold increase in the risk of HBV infection (OR =4.0; 95% CI = 1.8–9.0). Adjusting for age and sex did not modify the results. Within the Department of Dentistry, 15 subjects were HBsAg positive but anti‐HBc negative; 12 subjects were retested 1 year later and did not present any markers of past or current HBV infection. These results confirm the increased risk of HBV infection among hospital workers and suggest the presence of HBV variant(s) in Senegal.
Maturation of immunoglobulin G avidity after rubella vaccination studied by an enzyme linked immunosorbent assay (Avidity‐ELISA) and by haemolysis typing
Authors: Hedman Klaus , Hietala Jukka , Tiilikainen Anja , Hartikainen‐Sorri Anna‐Liisa , Räihä Kirsti , Suni Jukka , Väänänen Pertti , Pietiläinen Marjatta
Published: March 2007

Two tests were introduced recently for assessment of the avidity of rubella immunoglobulin antibodies. In the quantitative test—avidity—enzyme linked immunosorbent assay (ELISA)—IgG antibodies obtained from individuals shortly after primary infection with rubella virus are distinguished from those with past immunity by their antigen‐elution characteristics. This method uses agents that disrupt hydrophobic bonds in proteins [Kamoun PP (1988): Denaturation of globular proteins by urea: Breakdown of hydrophobic bonds? Trends in Biological Sciences 13:424–425.]. In the semiquantitative, presumptive test—haemolysis typing—the lowavidity rubella‐IgG antibodies are distinguished from the high‐avidity antibodies by the quality of their haemolytic zones in a radial haemolysis test. In the present study, both tests were applied to sera taken before and after vaccination with two different strains (Cendehill or RA 27/3) of live attenuated rubella virus. It was found that after vaccination of previously nonimmune subjects, IgG synthesized during the first 2 months had a very low avidity; IgG avidity increased dramatically during the subsequent 4 months and less markedly between 6 and 12 months after vaccination. On the contrary, the initially high IgG avidity of previous immune vaccinees remained at an elevated level postvaccination. These results provide a basis for identification of recent primary rubella virus infections, or vaccination reactions, by the avidity of specific IgG and also for their separation from rubella reinfections.
Altered in vitro uptake of the radiolabelled antiviral imaging “probe” e‐5‐(2‐125iodovinyl)‐2′‐deoxyuridine following administration of acyclovir
Authors: Klapper Paul E. , Lewis Alfor G. , Cleator Graham M. , Templeton Phillip , Longson Maurice
Published: March 2007

Current developments in the use of radiolabelled antiviral drugs as specific “probes” for virus‐infected cells in vivo may allow the specific neuroradiological diagnosis of herpes encephalitis. As “blind therapy” with the antiviral drug acyclovir may precede specific neuroradiological diagnosis, the aim of the present study was to investigate whether or not acyclovir interferes with the uptake of the radioprobe E‐5‐(2‐125 lodovinyl)‐2′‐deoxyuridine (rlVDU) by virusinfected cells in vitro. Acyclovir treatment (0.1 to 10 m̈g/ml) was shown to increase initial radioprobe uptake by virus‐infected cells. However, with continued incubation in the presence of acyclovir, intracellular radioactivity decreased to a level not significantly different from that associated with noninfected cells. A mechanism to explain these results is proposed. It was concluded that concurrent acyclovir therapy could interfere with neuroradiological diagnosis using rlVDU, although this may not occur with all the candidate radioprobes currently under investigation.
Inactivation of human hepatitis b virus dna polymerase by pyridoxal 5′‐phosphate
Authors: Oh Sang‐Hwan , Park Yeon‐Hee , Woo Koo
Published: March 2007

Human hepatitis B virus (HBV) DNA polymerase activity was inhibited by pyridoxal 5′‐phosphate (PLP) specifically and noncompetitively with respect to deoxythymidine triphosphate (dTTP). NaBH4 reduction of PLP‐HBV core proteins resulted in the complete inactivation of HBV DNA polymerase, and PLP modification of the enzyme was thought to be mediated through Schiff‐base formation.

HBV DNA polymerase has a Michaelis constant (Km) of 0.31 m̈M for dTTP and an apparent inhibition constant (Ki) of 0.2 mM for PLP. Its inactivation and modification by PLP may be useful in the study of not only the reaction mechanism of catalysis, but also the physicochemical nature of the enzyme.
High prevalence of human papillomavirus infection and possible association with betel quid chewing and smoking in oral epidermoid carcinomas in taiwan
Authors: Chang Kuo‐Wei , Chang Che‐Shoa , Chou Ming‐Jer , Lai Koon‐Siew , Choo Kong‐Bung
Published: March 2007

Seventeen oral epidermoid carcinomas, three oral papillomas, and 17 normal gingival tissues were tested for the presence of human papillomavirus (HPV) types 6, 11, 16, and 18 sequences by Southern blot hybridization. Episomal HPV‐16 sequences in various amounts were detected in 76.4% of the oral carcinomas and in all three cases of papilloma. However, only one of the 17 normal tissues was HPV positive with an unknown type. None of the samples contained HPV‐6, ‐11, or ‐18 sequences. Examination of the habits of the patients showed that 59% of the patients were betel quid chewers and 82% were smokers. Thus, the concurrent incidence of HPV infection and betel quid chewing and/or smoking habits in oral carcinoma patients observed in Taiwan is consistent with the view that both viral and chemical factors may be involved in the process of carcinogenesis.
Animal model of ultraviolet‐radiation‐induced recurrent herpes simplex virus infection
Authors: Stanberry Lawrence R.
Published: March 2007

The development of a model of induced recurrent herpes simplex virus disease would facilitate studies of the mechanism(s) responsible for reactivation of latent virus. The guinea pig model of genital herpes is characterized by a selflimited primary infection, the establishment of a latent infection in sensory ganglia, and the subsequent development of spontaneous recurrent genital infections. This study reports that exposure of latently infected female guinea pigs to ultraviolet radiation results in the induction of recurrent genital herpes simplex virus infections in approximately 60% of animals. Furthermore, there was a dose‐response relationship between ultraviolet irradiation and ability to induce recurrent disease. This model should prove useful in further investigations of the pathophysiology of herpes simplex infections.
Comparison of three techniques for detection of respiratory viruses in nasopharyngeal aspirates from children with lower acute respiratory infections
Authors: Salomón Horacio E. , Grandien Mónica , Weissenbacher Mercedes C. , Avila María M. , Pettersson Carl‐Axel
Published: March 2007

A comparison of immunofluorescence (IF), enzyme‐linked immunosorbent assay (ELISA), and isolation in tissue culture (TC) for detection of respiratory viruses was performed on 496 nasopharyngeal aspirates from children under 5 years of age with lower acute respiratory infections who were receiving attention at three hospitals in Buenos Aires, Argentina. All samples were tested by the three methods for respiratory syncytial virus (RSV), influenza A and B, adenovirus, and parainfluenza 1 and 3. Viral diagnosis was made in 167 samples (33.7%); of these, 124 (74.3%) were isolated in TC, whereas 120 (71.8%) were detected by ELISA and 127 (76%) by IF. RSV was detected in 121 samples, mainly by ELISA and IF. The sensitivity and specificity of each rapid technique as compared with isolation in TC were similar, reaching 98% and 92%, respectively. When ELISA was compared with IF, the sensitivity was 95%, and the specificity was 98%. Adenovirus was detected in 18 patients by TC. For this virus, rapid techniques sensitivity as compared with TC was low (almost 22%). Parainfluenza 3 was readily detected by IF and TC; influenza A, B and parainfluenza 1 were detected in few samples; and tissue culture proved more efficient than rapid techniques. The results indicate that both rapid techniques are good tools for the detection of most respiratory viruses except for adenovirus, for which TC cannot be omitted.
Hbv‐Dna hybridization in hepatocellular carcinoma associated with alcohol in japan
Authors: Horiike Norio , Michitaka Kojiro , Onji Morikazu , Murota Toshiko , Ohta Yasuyuki
Published: March 2007

To clarify the role of the hepatitis B virus (HBV) in hepatocellular carcinoma (HCC) associated with alcohol consumption, HBV‐DNA in the liver of 19 patients with HCC were investigated. HBV‐DNA was examined by Southern blot hybridization. HBV‐DNA was integrated into tumor cells from five out of six (83%) patients with HCC associated with HBs antigen (HBsAg)‐positive posthepatitic liver cirrhosis (LC), but this was not related to the history of alcohol intake. In 13 HCC patients of HBsAg‐negative alcoholic LC, HBV‐DNA integration was not detected in any patient. These findings suggest that HBV does not play a major role in the pathogenesis of HCC in HBsAgnegative alcoholics in Japan.
Hiv genome in peripheral blood mononuclear cells of seronegative regular sexual partners of hiv‐infected subjects
Authors: Pezzella M. , Vonesch N. , Sorice F. , Mannella E. , Mirolo M. , Miceli M. , Rosci M. A. , Morace G. , Rapicetta M. , Macchi B. , Angeloni P.
Published: March 2007

We have investigated the presence of the human immunodeficiency virus (HIV) by using in situ hybridization on peripheral blood mononuclear cells (PBMCs) from seronegative regular sexual partners of HIV‐infected subjects. The cells were hybridized with a 9 kilobase (kb) Sstl‐Sstl lambda BH 10 probe, which was able to recognize both viral mRNA and proviral cDNA. Labeling was done by chemical insertion of an antigenic sulfone group in cytosine moieties and was visualized by a double‐antibody immunohistochemical reaction. In all the subjects studied, the HIV genome was present. The HIV infected cells showed morphological aspects consistent with that of lymphocytes and monocytes. Our data suggest that the anti‐HIV seronegative individuals who are regular sexual partners of HIV‐infected subjects may be HIV‐infected.
Case report: Acute type a hepatitis presenting with hypotension, bradycardia, and sinus arrest
Authors: Gordon Stuart C. , Patel Atulkumar S. , Veneri Robert J. , Keskey Kristin A. , Korotkin Steven M.
Published: March 2007

The cardiovascular complications of viral hepatitis are not widely recognized. Profound hypotension and a variety of dysrhythmias, including sinus arrest, have been reported in association with viral hepatitis, but most of these cases were described before the advent of specific viral serologic markers. Furthermore, previous instances of cardiovascular complications occurred during the course of an established viral syndrome. We report the cases of two previously healthy women who presented with unexplained hypotension and bradycardia as the initial manifestations of acute icteric hepatitis A. This phenomenon appears to represent a variant clinical presentation of a common viral entity.
Enhanced endothelial cytopathogenicity induced by a cytomegalovirus strain propagated in endothelial cells
Authors: James Waldman W. , Sneddon Jeffrey M. , Stephens Ralph E. , Roberts William H.
Published: March 2007

Endothelial involvement has been implicated in cytomegalovirus (CMV) infection, a source of major complications in immunosuppressed individuals (e.g., those with acquired immune deficiency syndrome [AIDS] and organ transplants). Traditionally, CMV has been grown in fibroblasts; however, propagation in these cells may alter characteristics of the virus. In developing an in vitro model system of CMV/endothelial cell interaction, we have addressed this issue by propagating a clinical isolate, CMV VHL 1, in human umbilical vein endothelial (HUVE) cells by serial cocultivation of heavily infected cultures with fresh HUVE monolayers and have compared its infectious properties with those of the fibroblast‐raised strain, CMV AD169. In situ hybridization using a biotinylated DNA probe, as well as immunofluorescent staining for CMV specific antigen, has confirmed infection of HUVE cells inoculated with either strain of the virus. Infection of HUVE by VHL was accompanied by dramatic cytopathology not observed in AD169‐infected cells. Plaque assay of culture supernatants revealed greater virus production in VHL‐infected HUVE as compared with equivalently inoculated fibroblasts. In contrast, AD169 production in inoculated fibroblasts exceeded that in HUVE. These studies demonstrate the suitability of cultured endothelial cells as a substrate for CMV propagation and suggest that a strain of virus thus propagated may offer an accurate model of CMV/endothelial cell interaction in human disease.
New cd4( +) cell line susceptible to infection by hiv‐1
Authors: Tremblay Michel , Rooke Ronald , Geleziunas Romas , Wainberg Mark A. , Sullivan Arthur K. , Tsoukas Christos , Gilmore Norbert , Shematek Gene
Published: March 2007

Infection of a newly described human T lymphoid cell line, CEM‐CL10, with three different variants of HIV‐1 resulted in cytopathic effects followed by cell lysis. Following primary lytic infection, proviral DNA could not be detected by Southern blot analysis in the outgrowth of the surviving CEM‐CL10 cells 60 days after initial exposure to HIV‐1. These surviving cells could be further grown as a separate line, derived from CEM‐CL10, and were found to be resistant to subsequent infection by HIV‐1. A marked decrease in CD4 antigen expression was observed in these latter cells but not of the CD3 and transferrin receptor antigens. This decline in cell surface CD4 expression was correlated with both an absence of specific CD4 mRNA and with changes in structure of the CD4 gene. Both the HIV‐1‐sensitive CEM‐CL10 cell line and its CD4(‐), HIV‐1‐resistant derivative line, will be made available to interested investigators.
Postpartum subsidence of hepatitis B viral replication in hbeag‐positive carrier mothers
Authors: Lin Ho‐Hsiung , Chen Pei‐Jer , Chen Ding‐Shinn , Sung Juei‐Low , Yang Kung‐Ho , Young Yiang‐Chin , Liou Yu‐Sen , Chen Yuan‐Ping , Lee Tzu‐Yao
Published: March 2007

To elucidate the effects of pregnancy and delivery on hepatitis B e antigen (HBeAg)‐positive carrier mothers, 31 HBeAg‐positive carrier mothers were followed‐up postpartum 1 year, with 30 HBeAg‐positive nonpregnant female carriers as controls. Serum hepatitis B surface antigen (HBsAg), HBeAg titer, and hepatitis B virus (HBV)‐DNA concentration were studied at defined intervals. The results revealed that in the control group HBeAg titers and HBV‐DNA concentrations fluctuated, whereas the HBsAg titers showed little change, but HBeAg clearance or seroconversion to anti‐HBe were not noted on follow‐up. In contrast, one carrier mother sero‐converted to anti‐HBe during pregnancy and the antibody persisted thereafter. Five of the remaining 30 carrier mothers cleared HBeAg postpartum, and among these five cases, one also seroconverted to anti‐HBe. In addition, in another five of the 30 cases, the HBV‐DNA fell to undetectable level (< 0.04 ng/ml). All these ten cases had a common tendency of showing a decrease in HBeAg titers and/or HBV‐DNA concentrations 1–2 months after delivery. The HBeAg titers and HBV‐DNA concentrations in the other 11 cases remained unchanged, whereas the remaining nine cases had increased levels. It is concluded that subsidence of HBV replication is precipitated by delivery in one‐third of HBeAg‐positive carrier mothers in Taiwan, and this occurs most frequently 1–2 months postpartum.
Combined effects of acyclovir and human interferon‐α on herpes simplex virus replication in cultured neural cells
Authors: Hanada Naoki , Kido Shinji , Kuzushima Kiyotaka , Goto Yasuhiro , Rahman Mizanur M. , Morishima Tsuneo
Published: March 2007

The combined effects of Acyclovir [9‐(2α‐hydroxyethoxymethyl)guanine; ACV] and human interferon‐α (IFN‐α) on replication of the herpes simplex virus type I (HSV‐1) were determined in human neural cell lines, neuroblastoma (IMR), glioblastoma (118MGC), and glioma (U251MG). HSV‐1 grew well in all these cells, with final yields of more than 1 x 106 PFU/ml. In terms of virus‐yield reduction, ACV was found to be highly effective in IMR, moderately effective in U251MG, but ineffective in 118MGC. By contrast, IFN‐α reduced the virus yield significantly in 118MGC and in U251MG, but did not in IMR. Combined application of ACV and IFN‐α strongly inhibited the virus replication in all three cell lines with various degrees of synergism or additive effect. These results were also confirmed by immunofluorescent examinations. The sensitivity of HSV‐1 to ACV or IFN‐α was found to be different among the three different cell types. By combining the two agents, the virus growth was strongly suppressed in all the cells. These results suggest the importance of combination therapy for severe type of herpes simplex encephalitis in clinical practice.
DNA dot‐blot hybridization implicates human papillomavirus type 11‐DNA in epithelioma cuniculatum
Authors: Knobler R. M. , Schneider S. , Neumann R. A. , Bodemer W. , Radlwimmer B. , Aberer E. , Söltz‐Szöts J. , Gebhart W.
Published: March 2007

Epithelioma cuniculatum is a rare, low‐grade squamous cell carcinoma, belonging to the group of verrucous carcinomas. Evidence is presented that suggests that human papilloma virus type‐11 may be involved in the pathogenesis of epithelioma cuniculatum. HPV‐DNA was detected by dot‐blot hybridization with 32P‐labelled probe DNA. Plasmids containing HPV‐DNA were isolated by the benzoylated naphthoylated DEAE cellulose method (BND‐method), an improved procedure for routine detection of HPV‐DNA.
Restriction fragment differences between the genomes of the Oka varicella vaccine virus and american wild‐type varicella‐zoster virus
Authors: Adams Susan G. , Dohner Dennis E. , Gelb Lawrence D.
Published: March 2007

The Oka vaccine strains of varicella‐zoster virus (VZV) have a significantly different BgII DNA restriction pattern from that of American wild‐type isolates of VZV. This difference consists primarily of an additional BgII site, which lies within the BamHI “D” fragment. In conjunction with a study of the efficacy of an experimental Merck/Oka VZV vaccine, the area of the genome from which the most marked restriction pattern alteration arises was studied more closely to determine if there are other significant differences between the Oka strains and American wild‐type strains. BamHI “D” fragments from the DNA of the Oka parent strain (the progenitor of the vaccine strain), the RIT/Oka vaccine strain (a derivative of the Oka parent strain), the Merck/Oka vaccine strain, and the EF strain (an American wild type), were submitted to extensive endonuclease digestion studies to ascertain if additional unique restriction sites are present in the Oka parent or vaccine strains. The extra BgII restriction site characteristic of the Merck/Oka vaccine strain is also present in the DNA of the parent virus as well as its derivitives and was therefore not produced by the “attenuation” process. No other novel sites were found in the Oka parent or Oka‐derived strains in this section of the genome. The Merck/Oka vaccine strain of VZV, despite its Japanese origin, is therefore quite similar to circulating American varicella‐zoster virus strains. Varicella‐zoster virus DNA, at least in the area of the BamHI D fragment, also appears to be remarkably stable from strain to strain.
Hepatitis delta virus infections in intravenous drug abusers with hepatitis B in the west of Scotland
Authors: McCruden E. A. B. , Follett E. A. C.
Published: March 2007

The prevalence of delta virus infection was studied in 264 intravenous drug abusers (IVDAs) whose sera were found to be HBsAg positive between 1985 and 1987 and in 15 IVDAs from the period 1971–75, seven from 1976, 15 from 1979, and 37 from 1982. Delta markers were present in 41% of IVDAs with acute hepatitis B and in 65% of those who were chronic HBsAg carriers between 1985–87. The first evidence of delta virus infection was found in 1975. In 1976, 1979, and 1982, respectively, 42, 63, and 5% of IVDAs with acute hepatitis B had delta coinfection.

In the West of Scotland, delta infection has been established in the IVDA population since 1975. The very high percentage of IVDA carriers superinfected with the delta virus implies that there will be an excess of patients presenting with severe liver disease in the future.
Detection of human coronavirus 229E in nasal washings using RNA:RNA hybridization
Authors: Myint S. , Siddell S. , Tyrrell D.
Published: March 2007

A method is described for the detection of human coronavirus 229E (HCV 229E) in nasal washings using RNA:RNA filter hybridization. Volunteers were inoculated with HCV 229E, and daily nasal washings were collected. These washings were then examined for the presence of viral RNA using a single‐stranded RNA probe. Nucleic acid hybridization is shown to be a sensitive technique for the diagnosis of HCV 229E infections.
Comparison of different methods for detecting human immune deficiency virus in human immunodeficiency virus‐seropositive hemophiliacs
Authors: Schneweis K. E. , Ackermann A. , Friedrich A. , Kleim J.‐P. , Kornau K. , Ruff R. , Siefer‐Wippermann B.
Published: March 2007

Since the detection of antibodies against the human immune deficiency virus (HIV) does not definitely prove HIV infection in hemophiliacs, virus detection was attempted by virus isolation from the peripheral blood monocytes (PBL), by demonstration of p24 antigen and decline of p24 antibody, and by detection of viral DNA by the polymerase chain reaction (PCR). Virus isolation was optimized by immediate coculture of PBL and by replacement of the reverse transcriptase test by the p24 antigen test, whereas the elimination of CD8+ lymphocytes proved to be unnecessary. Virus detection was dependent on the clinical stage of the illness. Virus isolation in 70 of 211 patients (33%) was more sensitive than detection of p24 antigen or decline of p24 antibody. PCR was performed in 25 patients and indicated infection in all of 15 isolation‐positive cases and in 6 of 10 patients from whom virus was not isolated. Changes from negative to positive virus culture and from a weakly fusiogenic to a highly fusiogenic isolate were often accompanied by a progression of the disease. The results suggest that reactivation of HIV occurs when immune deficiency has become manifest. Apparently virus isolation detects only the virus already reactivated in vivo, whereas the PCR may also detect latent virus.
Delta hepatitis in kiribati: A pacific focus
Authors: Tibbs C. J.
Published: March 2007

From the Pacific Republic of Kiribati, 90/130 (69%) of hepatitis B‐infected individuals carry antibodies to hepatitis delta virus. The prevalence of delta infection varied between geographically isolated subgroups in the population and delta antibodies were found more frequently in subjects over the age of 10. It is unlikely that delta superinfection in this population is associated with an acute hepatitic illness. The prevalence of delta infection is the highest recorded from the Pacific region and thus migrants from Kiribati may have provided the source of infection in other populations in this region.
Passive protection of mice, goats, and monkeys against Japanese encephalitis with monoclonal antibodies
Authors: Zhang Mingjie , Wang Meixan , Jiang Shaozhun , Ma Wenyu
Published: March 2007

Six monoclonal antibodies (McAbs) against Japanese encephalitis virus (JEV) were tested for passive protection in JEV‐infected mice, goats, and rhesus monkeys. mG9 and nG2 had no protective effect; mG3 and 2D2 had some protective effect, but not sufficient to be of therapeutic significance; and 2H4 and 2F2 had excellent protective efficacy in mice even 120 hr after infection when most of the mice in the virus control group were sick. The mixture of 2H4, 2F2, mC3 (M‐McAb), and their F(ab')2 fragments showed excellent protection in mice, goats, and monkeys and was safe. The protective effects of McAbs correlated with their neutralization titers, but cytotoxicity‐mediated activities also played a role in protection.
Active immunization of homosexual men using a recombinant hepatitis B vaccine
Authors: Hess Georg , Rossol Siegbert , Voth Rita , Cheatham‐Speth Darline , Clemens Ralf , Zum Büschenfelde Karl‐Hermann Meyer
Published: March 2007

Twenty homosexual men [13 anti‐human immunodeficiency virus (HIV)‐positive, seven anti‐HIV negative] without HBsAg, anti‐HBs, and anti‐HBc were vaccinated with three 20 μg doses of a recombinant hepatitis B vaccine. All anti‐HIV‐positive homosexuals were nonresponders independent of the initial number of CD4‐positive cells. Among seven anti‐HIV‐negative individuals, five responded. After three doses of the vaccine, CD4‐positive cells fell in anti‐HIV positive individuals by 22.4%. A similar fall in CD4‐positive cells of an average 24.9% was noted in 17 matching, but nonvaccinated, anti‐HIV‐positive homosexuals. The study indicates that the efficacy of vaccination in anti‐HIV‐positive individuals is questionable. There is, however, no evidence that vaccination against hepatitis B might be harmful to anti‐HIV‐positive subjects.
Direct detection of human cytomegalovirus in urine specimens from renal transplant patients following polymerase chain reaction amplification
Authors: Olive D. Michael , Mufti Siham Al , Simsek Mehmet , Fayez Hana , Nakib Widad Al
Published: March 2007

A polymerase chain reaction (PCR) assay was used to amplify human cytomegalovirus (HCMV) directly from urine specimens taken from renal transplant patients. In serial urine samples from patients who had at least one specimen positive for HCMV; the PCR assay consistently detected the presence of HCMV DNA sequences, whereas virus detection by other tests such as enzyme‐linked immunosorbent assay (ELISA), nonradioactive DNA hybridization assay, and virus isolation were variable. Of 37 specimens positive by PCR, 36 were positive by either ELISA, hybridization assay, or virus isolation. Infectious virus was detected in 13 of the 37 PCR‐positive urines. HCMV DNA was detected by PCR in all samples that were positive for HCMV by either hybridization assay or virus isolation. The viral genome copy number was determined by PCR assay for several urine samples that were positive by virus isolation but negative for HCMV by ELISA or hybridization assay. Viral genome copy number estimates indicated the presence of HCMV at very low levels in these urines verifying the fidelity of the virus isolation procedures. The consistency of the PCR assay makes it an ideal method for detection of infection and monitoring antiviral drug therapy in patients infected with HCMV.
Evaluation of a new enzyme‐linked immunosorbent assay (ELISA) in the diagnosis of rhinovirus infection
Authors: Al‐Nakib W. , Dearden C. J. , Tyrrell D. A. J.
Published: March 2007

This study describes the evaluation of a newly developed ELISA for the direct detection of rhinovirus antigens in nasal washings. Of 54 volunteers inoculated with 100 TCID50 of human rhinovirus type 2 (HRV‐2), 50 (96.6%) and 32 (59%) excreted antigen and virus on at least 1 of 3 days investigated, respectively. Thirty‐three (61%) had significant rises in rhinovirus‐specific IgA by ELISA. Twelve (22%) developed symptoms of colds. Generally the ELISA detected antigen more frequently in volunteers later in the course of infection and provided evidence of infection in a higher proportion of asymptomatic compared with symptomatic volunteers. On the other hand, virus isolation detected virus more frequently earlier in the course of infection and in a higher proportion of symptomatic compared with asymptomatic volunteers. We conclude that rhinovirus antigen detection by ELISA is a simple, rapid, sensitive, and practical test to diagnose a rhinovirus infection and potentially a viable alternative to virus isolation.
Detection of supercoiled hepatitis B virus DNA and related forms by means of molecular hybridization to an oligonucleotide probe
Authors: Lin Hsiang Ju , Chung Hau Tim , Lai Ching Lung , Leong Susan , Tam Oi Sin
Published: March 2007

A novel assay for supercoiled and other fully double‐stranded forms of hepatitis B virus (HBV) DNA in blood is presented that utilizes molecular hybridisation to a radiophosphorous‐labeled oligonucleotide probe. The probe [5′‐d(ACGTGCA‐GAGGTGAAGCGA)] is complementary to the S(+)‐strand sequence furthest downstream, at the end of the gap. We examined blood specimens from 137 healthy HBsAg‐positive individuals, applying the probe to dots representing 2–3.5 ml serum or plasma. We found that supercoiled HBV is present in many HBV DNA‐positive blood specimens albeit in small quantities. Of the 104 specimens that were positive for HBV DNA of any form, 53 annealed to the probe. Serial specimens from the same subject taken over a period of months showed that the proportion of supercoil to other HBV DNA forms was variable. The presence of supercoil HBV DNA was not closely correlated with the level of serum HBV DNA polymerase. The supercoil is an HBV DNA form that can persist in the liver in the presence or absence of other replicative intermediates. This assay may enable further characterization of the status of HBV infection.
Immunoglobulin isotypes of anti‐HBc and anti‐HBe and hepatitis B virus (HBV) DNA elimination in acute hepatitis B
Authors: Sällberg Matti , Norder Helen , Weiland Ola , Magnius Lars
Published: March 2007

Antibodies to hepatitis B core antigen (anti‐HBc) and e antigen (anti‐HBe) were assayed in 46 sera from ten patients with acute hepatitis B utilizing immunoglobulin class‐ and subclass‐specific enzyme immunoassays (EIAs). The sera were sampled 1 to 512 days after onset of hepatic symptoms. Four patients cleared HBsAg rapidly, within 24 days, and six patients cleared HBsAg slowly, within 27–74 days after the onset of symptoms. In three of the patients with rapid clearance of HBsAg, hepatitis B virus (HBV) DNA was not detected in sera tested during the first week after onset. The fourth patient was not tested until 12 days after onset and was then found to be negative for HBV DNA. In four of the patients with slow clearance of HBsAg, HBV DNA was present during the first week of illness. In the other two patients, HBV DNA was not detected in the first serum, 11 and 17 days after the onset of illness. Anti‐HBc IgM and IgA1 were detected in all patients, with maximum titers shortly after onset. Anti‐HBc IgG1 was present in all sera tested. Anti‐HBc IgG2 was not detected in any of the sera. Anti‐HBc IgG3 and IgG4 were detected in all patient sera, with IgG3 paralleling IgG1, and IgG4 mainly in sera long after onset. Anti‐HBe IgG1, IgG3, and IgG4 were detected in three, two, and two patients, respectively. Anti‐HBe IgG2, IgM, IgA1, or IgA2 was not found in any patient. The time required for maximum titer of anti‐HBc IgG1 was shorter in the patients with rapid clearance of HBsAg. Thus, in these patients, the geometric mean titer (GMT) of anti‐HBc IgG1 had not increased significantly, in sera sampled 56–182 days after onset, whereas, in the patients with slow clearance of HBsAg, the increase in GMT of anti‐HBc IgG1 was significant (P < 0.05; Student's t test). The humoral responses to HBcAg and HBeAg in acute hepatitis B differ in that HBcAg elicits a vigorous IgM or IgA1 response whereas HBeAg elicits only low levels of IgG, mainly IgG1. Thus our findings support differences regarding T‐cell dependence, when comparing the humoral responses to HBcAg and HBeAg.
Natural infection of duck embryos with duck hepatitis B Virus: Time and tissue sequence of infection
Authors: Dixon Robert J. , Jones Noel F. , Freiman John S. , Wilson Naomi S.
Published: March 2007

There have been no studies addressing the detailed sequence of embryonic infection with duck hepatitis B virus (DHBV). Therefore, duck embryos from flocks infected with DHBV were examined to study the sequence of infection by DHBV in various embryonic tissues. Embryos from flocks infected with DHBV were harvested in duplicates from 7 to 25 days of incubation. Whole embryos (to 12 days) or dissected embryonic tissues were fixed, paraffin embedded, and stained for DHBV surface antigen (DHBsAg) using a peroxidase—antiperoxidase technique. Isolated hepatic cells were infected in 7‐day‐old embryos, and these increased in number until 11 days, when most cells were positive for DHBsAg. Endocrine pancreatic cells were positive from day 10, but only an occasional exocrine pancreatic cell was infected after day 20. Renal tubule cells were positive for DHBV by day 11, increasing in number until about day 18, after which a decline in numbers of infected cells occurred. Renal glomeruli became positive for DHBsAg from day 24. When present in the developing embryo, thymus, bursa of Fabricius, spleen, bone marrow, lung, and duodenum remained negative for DHBsAg. It was concluded that the timing of infection of specific tissues was not necessarily related to cellular maturity but may reflect a need for specific metabolic functions that permit viral replication.
Antibody response to herpes simplex virus glycoproteins gB and gD
Authors: Bernstein D. I. , Frenkel L. M. , Bryson Y. J. , Myers M. G.
Published: March 2007

The antibody response to herpes simplex virus (HSV) glycoproteins B and D was evaluated using these cloned glycoproteins in an ELISA assay and compared to a standard Western blotting procedure. The ELISA assay appeared to be more sensitive for detecting gB and gD antibodies. Antibodies to gB but not gD were detected in the acute sera of patients presenting with their first episode of true primary genital herpes. The geometric mean HSV gB titer (log 2) was also significantly higher than the geometric mean gD titer in the convalescent sera of these patients (7.9 ± 0.7 vs. 5.5 ± 0.9, P < .003). A cross‐reaction between HSV gB and varicella zoster virus (VZV) gp II was demonstrated. Thus, it is possible that a previous VZV infection could prime the immune system to respond rapidly and more vigorously to HSV gB. Indeed, in this report we demonstrated a significant correlation between the VZV ELISA absorbance and the titer to HSV gB and also detected a higher VZV ELISA absorbance and HSV gB titer in the acute sera of patients with a true primary HSV infection compared to other HSV seronegative VZV seropositive patients. Use of this quantitative assay should allow further investigation into the relationship of the immune response to these important targets and the clinical course of HSV disease.
Polymerase chain reaction for studies of mother to child transmission of HIV1 in Africa
Authors: Paterlini P. , Coeur S. Lallemant‐Le , Lallemant M. , M'Pelé P. , Dazza M. C. , Terre S. , Moncany M. , Jourdain G. , Courgnaud V. , N'Zingoula S. , Larouzé B. , Griscelli C. , Bréchot C.
Published: March 2007

The feasibility and implications of the use of the polymerase chain reaction (PCR) assay in studies of HIV1 mother to child transmission in Africa were investigated. Uncultured leukocyte blood cells (PBL) obtained in Brazzaville (Congo) from newborns and infants (mean age = 27 weeks) of infected mothers were tested. HIV1 DNA sequences were identified in the PBL of six of eight newborns and 14 of 23 babies born to HIV1‐positive mothers. In addition two of four babies, who at birth had been seropositive and subsequently were seronegative, were HIV1 DNA positive by PCR. This study demonstrates directly, therefore, a high rate of HIV1 transmission in Africa; it also indicates that PCR should be used for such epidemiological studies.
Antibodies to polioviruses in an Israeli population and Overseas volunteers
Authors: Margalith M. , Morag A. , Fattal B.
Published: March 2007

A seroepidemiological survey of prevalence of antibodies to poliovirus types 1–3 was carried out in 1,800 residents of 30 agricultural settlements (kibbutzim) in Israel during 1980 and 1981.

The survey showed that in 1980 84%, 90%, and 79% of the kibbutz residents exhibited antibodies to poliovirus types 1–3, respectively. In 1981 the figures were 81%, 91%, and 73%. Significantly lower levels of antibodies for poliovirus type 3 in 1980/1981, and for poliovirus type 1 in 1981, were noticed in the age group 6–17 years compared with the age group of > 18 years. Infants and young children of the kibbutzim received the Salk polio vaccine during 1957–1961 and the Sabin oral polio vaccine since 1961.

Among overseas volunteers from Western countries who came during 1980 and 1981 to work in the kibbutzim for a limited period of time, 88% exhibited antibodies to poliovirus type 1, 94% to poliovirus type 2, and 75% to poliovirus type 3.

No significant differences in the level of antibodies were detected between males and females or between residents in those kibbutzim using or not using wastewater for irrigation.
The absence of hepatitis B virus DNA in hepatitis B e antigen positive sera from chronic hepatitis B surface antigen carriers in China
Authors: Scott Janet S. , Pan Pei‐En , Pace Raymond A. , Sloots Theo P. , Cooksley W. Graham
Published: March 2007

Sera from 20 Chinese patients with chronic hepatitis B were examined for hepatitis B e antigen and hepatitis B virus (HBV) DNA. There was considerable discordance with HBV DNA not being detectable in 10 out of 13 (77%) patients who were hepatitis B e antigen positive. Further testing for anti‐HBe and HBV‐DNA polymerase activity confirmed the results. Possible reasons for this discordance are discussed but neither hepatitis D (delta) infection nor the acquired immunodeficiency syndrome (AIDS) could be implicated.
Molecular characterization by RNA‐RNA hybridization of a serotype 8 human rotavirus with “super‐short” RNA electropherotype
Authors: Ohshima Atsushi , Takagi Toshiaki , Matsuno Toyoko Nakagomi Shigeo , Matsuno Shigeo , Nakagomi Osamu
Published: March 2007

A human rotavirus isolate (the 69M strain) with a “super‐short” RNA electropherotype has been classified as serotype 8, a new human serotype [Matsuno et al., 1985] RNA‐RNA hybridzation using 32P‐labeled transcription probes was used to assess the genetic relatedness of the 69M strain to a panel of human and animal rotaviruses. The 69M strain showed a medium level of homology with subgroup I, serotype 2 human rotaviruses with short RNA electropherotypes. However, the 69M strain was not significantly related to any other human rotavirus strains tested in this study, including recently identified serotype 9 strains (represented by the WI61 strain) and subgroup I human rotaviruses with long RNA electropherotypes (represented by the AU‐1 strain) that are shown to be genetically distinct from other human rotavirus strains. Unexpectedly, a medium level of homology was also found between the 69M probe and the double‐stranded RNAs from bovine rotavirus strains (represented by the NCDV strain).
An epidemic of respiratory syncytial virus in elderly people: Clinical and serological findings
Authors: Agius G. , Dindinaud G. , Biggar R. J. , Peyre R. , Vaillant V. , Ranger S. , Poupet J. Y. , Cisse M. F. , Castets M.
Published: March 2007

In 1984–1985, an outbreak of respiratory syncytial virus (RSV) infection occurred in two geriatric wards. Among 68 patients (mean age ± SD = 82.5 ± 12.5 years) with respiratory signs, 52 had signs caused by RSV infection. Among all patients, the clinical and serological attack rates were 61.2% and 75.0%, respectively. The most frequent clinical presentation was intensive coughing (96.1%) and fever (96.1%) associated with expectorate (63.5%). The duration of the respiratory symptoms was 5 to 7 days. The disease gradually resolved, although in eight (15.4%) patients complications occurred. For periods of up to 1 year after infection, 172 sera were obtained and tested by complement fixation test (CFT), fluorescent assays for titrating specific IgG, IgA, and IgM, and Western blotting. Specific IgM appeared in six (11.5%) of the infected patients and peaked 2 to 6 months after infection, and there was no significant correlation with severity of clinical symptoms. However, higher peak G and A antibody responses were observed in persons with rales (CFT: P=0.008; IgG: P=0.042; IgA: P=0.020), cough (IgG: P=0.034), sputum (IgG: P=0.030), dyspnea (CFT: P=0.024), conjunctivitis (CFT: P=0.025), and bronchitis (CFT: P=0.018). The temporal patterns of IgA and CFT results were found to be similar, whereas IgG peaked later, i.e., between 2 and 6 months. The patients with the most severe symptoms had the highest antibody titers obtained by conventional tests and by Western blots. Thus, RSV can be an epidemic pathogen among elderly persons, although this illness is usually mild.
Persistence of hepatitis A virus in fulminant hepatitis and after liver transplantation
Authors: Fagan Elizabeth , Yousef Galal , Brahm Javier , Garelick Hemda , Mann George , Wolstenholme Adrian , Portmann Bernard , Harrison Tim , Mowbray James F. , Mowat Alexander , Zuckerman Arie , Williams Roger
Published: March 2007

A peroxidase‐labelled, specific mouse monoclonal antibody to hepatitis A virus (HAV) and an in situ hybridization technique (streptavidin‐biotin‐horseradish peroxidase reaction) with an HAV‐specific cDNA probe (recombinant plasmid pAWHA comprising 1.8 kb of the HAV‐specific cDNA, located toward the 3′ end of the genome) were used to detect HAV in liver tissues in two patients with fulminant viral hepatitis type A treated by liver transplantation after a protracted (day 40: case 1) and relapsing (day 60: case 2) clinical course. HAV antigens and HAV‐specific genomic sequences were detected in the hepatectomy tissues and in serial biopsies of the liver grafts through to final follow‐up at 2 months (case 2) or death at 7 months after re‐grafting for chronic rejection (case 1). In the fulminant liver parenchyma, numerous degenerating and some surviving hepatocytes were positive and randomly scattered. The immunoperoxidase staining was predominantly cytoplasmic and often granular. The localization of the cDNA probe was predominantly nuclear/perinuclear but was occasionally cytoplasmic. High‐titre IgM‐anti‐HAV antibodies persisted until death (case 1) or resolution (5 months) of an acute hepatitis (case 2), which occurred at 2 months, accompanied by HAV antigen (ELISA), in stool. Intact replicating virus particles must have been present in one or more sites in each case, including extrahepatic locations, with a viraemia as the most likely explanation for subsequent reinfection of the grafts.