bluuebunny/crossref_metadata_2025 huggingface dataset:
Expression of human papillomavirus types 16 and 18 DNA sequences in cervical carcinoma cell lines
Authors: Pater Mary M. , Pater Alan
Published: March 2007

A total of six cervical carcinoma cell lines, of which two contained human papillomavirus (HPV) type 16 and four contained HPV type 18 DNA, were examined for viral RNA expression by spot‐blot hybridization. All six cell lines contained HPV‐specific cytoplasmic poly(A)‐containing RNA. The same RNA species were detected in both cytoplasmic and polysomal fractions, indicating nonselective translation of these messages into protein products. Primer extension analysis of RNA from the cell lines revealed the presence of three major start sites at the 5′ end of the E6 gene for RNA from HPV 16‐containing cell lines and four major start sites at the 5′ end of the E6 gene for RNA from HPV 18‐containing cell lines. One cell line, ME 180, had a distinct pattern with only one minor HPV 18 start site. Northern‐blot hybridization using the E6‐E7 region of HPV 16 and 18 DNAs as probes revealed the presence of E6‐E7 transcripts in all the cell lines except ME 180. Southern‐blot analysis revealed that while sequences hybridizing to the E6‐E7 coding region were present, the 5′ end region containing the enhancer‐promoter region of viral DNA is deleted in ME 180. Subsequent Northern‐blot hybridization using the E1 region of HPV 18 DNA as probe revealed that the viral RNA in ME 180 is transcribed from this region.
Detection of EBV and HPV DNA sequences in oral “hairy” leukoplakia by in situ hybridization
Authors: Eversole L. R. , Stone C. E. , Beckman A. M.
Published: March 2007

Oral “hairy” leukoplakia (OHL) is a white lesion of the oral mucosa, usually located on the lateral tongue among human immunodeficiency virus (HIV) positive acquired immunodeficiency syndrome (AIDS)‐risk patients. The lesion has been reported to be associated with Epstein‐Barr virus (EBV) and human papillomavirus (HPV). Twenty surgical biopsy specimens were evaluated for the presence of HPV genus‐specific antigen, HPV 2/4, 6/11, and 16/18 DNA and EBV DNA by in situ hybridization employing formalin‐fixed paraffin‐embedded sections. Three cases exhibited immunoreactivity for HPV genus‐specific antigen, with localization in cytopathically altered upper spinous layer keratinocytes. HPV 16, 18, or related DNA sequences were identifiable in a single case. Alternatively, employing an EBV long internal repeat subgenomic probe, 19 cases were found to harbor EBV DNA. In all positive cases, the hybrids were localized to upper spinous layer keratinocytes exhibiting nuclear/cytoplasmic vesiculation. It is concluded that OHL is consistently associated with EBV; furthermore, viral replication, as evidenced by DNA localization, corresponds to ultrastructural evidence of capsid and envelope assembly in the more differentiated layers of oral epithelium.
Hepatocellular carcinoma: Possible etiologies in patients without serologic evidence of hepatitis B virus infection
Authors: Tabor Edward
Published: March 2007

Although hepatitis B virus (HBV) has been closely associated with the development of hepatocellular carcinoma (HCC), no serologic markers of HBV can be found in up to 11% of HCC patients in developing countries and up to 68% of HCC patients in industrialized countries. Despite the absence of HBV serologic markers in these HCC patients, HBV DNA sequences have been found to be integrated into HCC DNA in 13–100% of these patients, indicating a possible role of HBV in the etiology of their HCC. Although six patients with chronic non‐A, non‐B hepatitis virus infection who were followed have been documented to develop HCC, it is not known whether the non‐A, non‐B hepatitis viruses cause or contribute to the development of HCC in some HCC patients without HBV serologic markers. Ethanol, cigarette smoking, oral contraceptives, and aflatoxin also have been suggested as possible etiologies and should be studied further. Suggested etiologies that are not supported by the published data include alpha‐1‐antitrypsin deficiency and schistosomiasis.
Persistent productive HIV infection in EBV‐transformed B lymphocytes
Authors: Tozzi Valerio , Britton Sven , Ehrnst Anneka , Lenkei Rodica , Strannegård Örjan
Published: March 2007

The susceptibility to HIV infection of 14 B‐cell lines established from five healthy HIV seronegative and from six HIV seropositive subjects by lymphocyte transformation with EBV and/or by lymphocyte cultivation with cyclosporin A was studied. Although the cell lines contained different proportions of CD4‐positive cells, as shown by flow cytometry, all of them could be infected with the SF‐2 strain of HIV. Infection was blocked by a monoclonal antibody directed against the viral attachment site of the CD4 molecule, even in a line that lacked demonstrable CD4 receptors. B‐cell lines with high proportions of CD4‐expressing cells produced HIV p24 antigen more rapidly and at higher concentrations than cell lines with low CD4 expression. Although HIV infection resulted in some cytopathic effects, it was possible to cultivate the infected cells for more than 8 months without refeeding the cultures with uninfected cells. Even in long‐term cultures, there was a continuous production of infectious HIV, as detected by transfer of culture supernatants to other susceptible cell lines. The production of viral antigens was consistently more pronounced in the B‐cell line with the highest CD4 positivity than it was in a permissive T‐cell line (HUT‐78) infected in the same manner. These results indicate that HIV can chronically and productively infect transformed B cells via interaction with CD4 molecules. Thus it is possible that B cells may constitute a source of infectious virus in HIV‐infected EBV‐positive individuals.
False‐positive sera do not react with human immunodeficiency virus (HIV) Gag‐encoded recombinant antigen
Authors: Bukrinsky M. I. , Syrtsev V. A. , Popov S. A. , Barsov E. V. , Chaplinskas S. A. , Karamov E. V.
Published: March 2007

Ten sera from healthy blood donors positive by enzyme‐linked immunoadsorbent assay (ELISA) were studied by immunoblot assay using natural and recombinant proteins. They interacted only with p17 or p24 proteins but were nonreactive with a recombinant protein (RP 50), which carries antigenic determinants to p17 and p24. Reactions were not blocked by preincubation of sera with genetically engineered p17 and p24 or purified viral p24, indicating that some new epitopes were formed during the Western blot procedure. Recombinant gag‐encoded protein is required for confirmation of human immunodeficiency virus (HIV) seropositivity.
Epidemiology of molluscum contagiosum using genetic analysis of the viral DNA
Authors: Scholz J. , Rösen‐Wolff A. , Bugert J. , Reisner H. , White M. I. , Darai G. , Postlethwaite R.
Published: March 2007

The molecular epidemiology of molluscum contagiosum virus (MCV) infections was investigated by restriction endonuclease analysis of the genomes of 222 separate isolates collected from 147 patients living in Germany (33 patients), Hong Kong (6 patients), and Scotland (108 patients). MCV type 1 (MCV‐1) caused 96.6% of the infections, and MCV type 2 (MCV‐2) caused 3.4%. However, isolates from four of the 142 MCV‐1‐infected patients and two of the five MCV‐2‐infected patients showed minor differences in their DNA restriction patterns because of the loss of a single or very few recognition sites for the enzymes used. No genome variations were detected amongst isolates collected from different sites or on several occasions from individual patients or from closely related patients. Southern blot hybridization revealed a high level of relatedness between MCV‐1 and 2. No differences were seen in the appearance or anatomical localization of lesions caused by either virus type. In particular, there was no preferred genital localization for MCV‐2 infections.
Early diagnosis of cytomegalovirus infection in renal transplant and dialysis patients by DNA‐DNA hybridisation assay
Authors: Agha S. A. , Mahmoud L. A. , Archard L. C. , Abd‐Elaal A. M. , Selwyn S. , Mee A. D. , Coleman J. C.
Published: March 2007

One hundred forty‐eight urine specimens were collected from 47 renal transplant and dialysis patients and screened for the detection of cytomegalovirus (CMV). Diagnosis of CMV infection was suggested in 17 out of 47 patients (36.2%) by more than one of the five methods used. DNA hybridisation assay (DNA HA) using 32P‐labelled probe detected CMV DNA in 15 (31.9%) of 47 patients, whereas virus isolation on conventional tube cell cultures (CTC), immunofluorescence incorporating monoclonal antibodies on centrifugation vial cultures (IF), complement fixation test (CFT), and electron microscopy (EM) yielded positive results in only nine (19.2%), 12 (25.2%), 11 (23.4%), and one (2.1%) of 47 patients, respectively. The significance of these results obtained by DNA HA lies not only in the apparent increase in number of patients diagnosed, but also in both early and rapid detection of CMV DNA. More importantly, the DNA HA is highly specific in that it correlates accurately with clinical and laboratory data characteristic of CMV disease. In respect of clinically manifest CMV disease, the specificity of DNA HA, CTC, IF, CFT, and EM was 87.5, 43.7, 56.3, 43.7, and 6.3%, respectively. These advantages of DNA HA make it the test of choice for early diagnosis of CMV infections in immunosuppressed patients.
High frequency of hepatitis B virus DNA in anti‐HBe positive sera on longitudinal follow‐up of patients with renal transplants and chronic hepatitis B
Authors: Norder Helene , Brattström Christina , Magnius Lars
Published: March 2007

Hepatitis B virus (HBV) markers were determined in 821 patients receiving renal allografts and undergoing immunosuppressive therapy during 1970–1986. Twenty‐four of the patients with a renal transplant functioning for longer than 1 year originally were or became chronic carriers of hepatitis B surface antigen (HBsAg). These patients remained carriers during the follow‐up period, which lasted until death or until the end of 1986. Follow‐up time was 1.2–15.3 years (mean 9.1 years). A total of 301 samples from the HBsAg‐positive patients were tested for HBV DNA and HBeAg/DNA and HBeAg/anti‐HBe.

Nine patients who were constantly positive for HBeAg also remained positive for HBV DNA. Reactivation of HBV replication occurred in 11 patients. Among these, HBV DNA and HBeAg varied in parallel in six patients, three patients developed anti‐HBe, and two patients were constantly positive for anti‐HBe. Another four of the 24 patients seroconverted to anti‐HBe, and two of these also lost HBV DNA.

Three of 12 deceased patients died from liver failure during follow‐up. None of these three had been constantly positive for HBeAg or HBV DNA, but they had had reactivations of HBV; two were also positive for HBV DNA in serum specimens available from their terminal month.

HBV DNA was demonstrated in 99% of HBeAgpositive and 53% of anti‐HBe‐positive sera and in at least two samples from each of the 24 patients. It may be presumed that the reason for failure to eliminate the HBV infection, and for progressive liver disease in this group of patients, was a discordant T‐and B‐cell response to HBeAg. Due to the high frequency of HBV DNA in HBeAg negative and in anti‐HBe positive sera from immunosuppressed HBsAg‐positive renal transplant patients, determination of serum HBV DNA is important for defining infectivity in these patients.
Protective effect of a synthetic peptide comprising the complete pres2 region of the hepatitis b virus surface protein
Authors: Emini Emilio A. , Larson Vivian , Ellis Ronald W. , Miller William J. , Gerety Robert J. , Conard Patricia , Garsky Victor M. , Anderson Charles A. , Eichberg Jorg , Rick Lee D.
Published: March 2007

A peptide was synthesized containing the entire 55 amino acid residue sequence of the hepatitis B virus (HBV) surface antigen preS2 region (ad subtype). The unconjugated peptide was inoculated into four chimpanzees. Following multiple injections, all of the animals developed specific antipeptide antibodies that reacted with intact surface antigen particles bearing the preS2 moiety. All four peptide‐inoculated animals were found to be protected from infection after intravenous challenge with live HBV of either the ad or ay subtypes.
Brief report: Primary human herpesvirus 6 infection in a patient following liver transplantation from a seropositive donor
Authors: Ward K. N. , Gray J. J. , Efstathiou S.
Published: March 2007

A virus, isolated from peripheral blood lymphocytes of a patient 23 days after liver transplantation, was identified by negative contrast and thin‐section electron microscopy, DNA hybridisation, and immunofluorescence as human herpesvirus 6 (HHV‐6). The patient was seronegative for antibodies to HHV‐6 before receiving the liver from a seropositive donor. Seroconversion for IgM and IgG antibodies occurred at the same time as the viraemia, indicating a primary response to HHV‐6 infection that was probably acquired from the donor liver. The relationship of the infection to the patient's subsequent disease episodes is discussed.
Immune response of patients with congenital coagulation disorders to hepatitis b vaccine: Suboptimal response and human immunodeficiency virus infection
Authors: Miller Elizabeth J. , Lee Christine A. , Holmes Sue , Kernoff P. B. A. , Thomas H. C. , Karayiannis P.
Published: March 2007

Seventy‐eight patients with congenital coagulation disorders were treated with hepatitis B vaccine either subcutaneously or intradermally. All the children (eight vaccinated subcutaneously and eight vaccinated intradermally) responded. Seventeen of 19 (90%) anti‐HIV‐negative adults vaccinated subcutaneously and 14/25 (56%) anti‐HIV‐negative adults vaccinated intradermally showed an immune response. At 24 months, the anti‐HBs level was > 10 IU/I in all children vaccinated subcutaneously, 83% of children vaccinated intradermally, 77% of adults vaccinated subcutaneously, and 55% of adults vaccinated intradermally. Eight of 15 (53%) adult patients who were anti‐HIV positive were also anti‐HBc positive before vaccination and 6/8 (75%) failed to produce an amnestic response to vaccine. Subcutaneous vaccination with regular monitoring of anti‐HBs levels and appropriate boosting is recommended.
Epidemiology of hepatitis B in eastern Kenya
Authors: Hyams Kenneth C. , Morrill John C. , Woody James N. , Okoth F. A. , Tukei P. M. , Mugambi M. , Johnson Bruce , Gray Gregory C.
Published: March 2007

A cross‐sectional survey of outpatients attending the three district hospitals in the towns of Mombasa, Kilifi, and Malindi was conducted to determine the patterns of hepatitis B transmission in eastern Kenya. Of 1,533 study subjects (mean age 21.9 ± 13.2 years; range, 4 months to 80 years), 11.4% were positive for HBsAg and 56.2% were seropositive for at least one hepatitis B marker (HBsAg, anti‐HBs, or anti‐HBc). Antidelta antibody was found in 1.2% of HBsAg‐positive samples. HBeAg was found in 36.0% of HBsAg‐positive samples obtained from women of childbearing age. The prevalance of seropositivity for hepatitis B markers was positively correlated with age, increasing from 20% in subjects less than 4 years old to more than 80% in study subjects greater than 40 years old. On multivariate analysis, male sex was found to be associated with HBsAg positivity, and age and previous deliveries of children were associated with seropositivity for any hepatitis marker (HBsAg, anti‐HBs, or anti‐HBc). An effective hepatitis B immunization strategy in this region of Kenya would require vaccination early in life because a major portion of hepatitis B transmission occurs in childhood.
Brief report: Killer cell defect and persistent immunological abnormalities in two patients with chronic active epstein‐barr virus infection
Authors: Joncas J. , Monczak Y. , Ghibu F. , Alfieri C. , Bonin A. , Ahronheim G. , Rivard G.
Published: March 2007

Two members of a family have manifested a syndrome of chronic active Epstein‐Barr virus (EBV) infection. A father and his daughter suffered prolonged or recurrent mononucleosis, with splenomegaly, anemia, and intermittent fever; persistent immunological abnormalities included defective natural killer (NK) cytotoxicity, inverted CD4/CD8 ratios, hyper IgG1, high EBV viral capsid antigen (VCA) and early antigen (EA) antibodies, and low or undetectable EBV nuclear antigen (EBNA) antibody titers. The EBV seronegative member of the family was free of these abnormalities. However, NK activity in the seronegative individual was low‐normal and its EBV‐specific antibody‐dependent K‐cell cytotoxicity (EBV‐ADCC) was abnormally low, suggesting that this K‐NK cell defect may be primary. The father, who suffered from the syndrome for more than 15 years, lacked (or lost) antibodies to EBV‐envelope and infected cell membranes, such as antibody‐dependent cellular cytotoxicity (ADCC), neutralizing (NT), and gp 350/220 antibodies. Slow improvement over a period of years was heralded by rising NK cytotoxicity.
Toga‐Like virus as a cause of fulminant hepatitis attributed to sporadic non‐a, non‐b
Authors: Fagan Elizabeth A. , Portmann Bernard , Williams Roger , Ellis David S. , Tovey George M. , Zuckerman Arie J. , Lloyd Graham
Published: March 2007

Virus‐like particles (60–70 nm) with spiked surfaces budding into cell vacuoles and rod‐shaped inclusions were detected in nuclei of hepatocytes from a British patient transplanted for sporadic non‐A, non‐B fulminant hepatitis (NANBFHF), probably contracted in Kenya. Identical particles were seen in two successive grafts (days 2 and 10) at regrafting for recurrent FHF. Ultrastructural features resembled those of the RNA‐containing arbovirus, Rift Valley fever virus, but serological markers against a representative panel for arboviruses (Togaviruses) and transmission in mice proved negative. The particles shared features with the different arboviruses seen in the hepatectomy specimen of a second patient with NANB‐FHF, and in both patients an insect vector was implicated in the clinical history. The particles were identical in size to those of a third patient with NANB‐FHF, who had remained in the United Kingdom. These findings, together with the recent report of isolation of an RNA‐containing virus resembling the Togaviridae, in parenteral NANB, suggest that several exotic virus‐like agents resembling the arboviruses may be involved in the aetiology of NANB, including in the sporadic forms of FHF in the United Kingdom.
Seroepidemiology of hepatitis b virus, hepatitis d virus, and human immunodeficiency virus infections among parenteral drug abusers in southern taiwan
Authors: Chung Dye‐Chii , Ko Ying‐Chin , Chen Chien‐Jen , Chen Eng‐Rin , Wu Chung‐Chieng , Wu Pyng‐Shouh
Published: March 2007

A total of 390 parenteral drug abusers (PDAs) at the Kaohsiung Municipal Narcotics Abstention Institute were examined for markers of hepatitis B virus (HBV), hepatitis D virus (HDV), and human immunodeficiency virus (HIV). All sera were tested for hepatitis B surface antigen (HBsAg), surface antibody (anti‐HBs), and core antibody (anti‐HBc) by radioimmunoassay (RIA) and for antibody to HIV (anti‐HIV) by enzyme‐linked immunosorbent assay (ELISA). Hepatitis B e antigen (HBeAg) and antibody to HDV (anti‐HDV) were also tested for HBsAg‐positive serum samples. Although the HBsAg‐positive rate (22.1%) among PDAs was similar to that of the general population in southern Taiwan, the HBV infection rate (99.2%) and the anti‐HDV‐positive rate (78.5%) among HBsAg‐positive subjects were significantly higher than those of the general population in southern Taiwan (P < 0.0001). None of the PDAs studied were positive for anti‐HIV.

The levels of serum glutamic oxaloacetic transaminase (SGOT) and serum glutamic pyruvic transaminase (SGPT) among PDAs were significantly higher than those of the general population in southern Taiwan (P < 0.0001). The more frequent the institutionalisation, the higher the infection rates with HBV and HDV and elevated levels of SGOT and SGPT. Horizontal transmission through parenteral drug abuse may be considered a possible reason for the significantly higher rates of HBV and HDV among parenteral drug abusers.
Mutations that change the immunological subtype of hepatitis b virus surface antigen and distinguish between antigenic and immunogenic determination
Authors: Ashton‐Rickardt Philip G. , Murray Kenneth
Published: March 2007

The molecular basis of the d or y immunological subtype of hepatitis B virus (HBV) surface antigen (HBsAg) has been investigated by mutation of specific amino acid residues. When combined with substitution of serine 113 by threonine, replacement of arginine 122 by Iysine or of tyrosine 134 by phenylalanine, or both of these changes, altered the antigenic subtype of HBsAg from y+ d– to y+ d+. These same mutations had a more dramatic effect on the subtype of antibodies induced by the antigens, a combination of all three mutations completely changing the subtype from y to d. Our study thus identifies residues in HBsAg that not only affect the subtype but discriminate between changes in antigenic and immunogenic behaviour. It also shows how the y and d subtypes may be manifest by the same molecule.
Free and antibody‐complexed antigen and antibody profile in apparently healthy HIV seropositive individuals and in aids patients
Authors: Portera Mario , Vitale Francesco , Licata Rosaria La , Alesi Domenico Russo , Lupo Giuseppina , Bonura Filippa , Romano Nino , Cuonzo Giordano Di
Published: March 2007

The pattern of free and antibody‐complexed HIV antigen and the antibody profile were investigated retrospectively in 305 serum samples taken from 22 AIDS patients before and during the development of AIDS and from 40 apparently healthy seropositive individuals.

Most AIDS patients were found positive for both free and complexed antigen and had high gp41 antibody titres but low or undetectable p24 antibody. Four different patterns of HIV antigenaemia were observed: 1) positive for both free and complexed antigen; 2) negative for free HIV antigen at first, but always positive for complexed antigen; 3) positive for free antigen without complexed antigen; and 4) negative for both free and complexed antigen.

The development of immune complexes preceded the appearance of free antigen and might reflect the ongoing viral replication with antigen excess and binding of anticore antibodies. No correlation was found between the development of AIDS symptoms and either the duration of free antigen positivity or the level of antigenaemia.

A different pattern was observed in apparently healthy seropositive individuals: 90% of whom had high antibody titres to p24 and gp41 and were persistently negative for free and complexed HIV antigen.

This study demonstrates that testing HIV markers in sequentially collected serum samples from HIV seropositive individuals is a useful and simple tool for early identification of persons at risk of developing AIDS.
Serum IgG subclass responses of humans to inactivated and live influenza A vaccines compared to natural infections with influenza A
Authors: Hocart Michael J. , Mackenzie John S. , Stewart Geoffrey A.
Published: March 2007

Studies with various viral agents have suggested that a preferential production of IgG subclasses may occur during infection, but limited information has been reported on the IgG isotypes produced during vaccination with live or killed virus preparations. The serum IgG subclass responses to influenza A infection or inoculation with live or killed influenza A vaccines were examined by an enzyme‐linked immunosorbent assay, and results were expressed using a 4‐parameter logistic model. It was observed that IgG1 was induced by both natural infections and the live virus vaccine depending on the dose given. Inactivated vaccines induced significant titres of IgG1, IgG2, and IgG3 isotypes in vaccines, again depending upon the amount of virus preparation administered.
Speed of progression to AIDS and degree of antibody response to accessory gene products of HIV‐1
Authors: Reiss Peter , Lange Joep M. A. , de Ronde Anthony , de Wolf Frank , Dekker John , Debouck Christine , Goudsmit Jaap
Published: March 2007

Antibodies to E. coli‐produced HIV‐1 nef, rev, tat, vpu, and vpr proteins were measured by enzyme immunoassay in serial sets of sera from 72 men seroconverting for antibodies to HIV‐1 structural proteins, and from 190 initially symptom‐free men who were seropositive for these antibodies at entry into the study. In the men seroconverting for antibodies to structural proteins the levels of nef‐, rev‐, and tat‐specific antibodies, but not of vpu‐, and vpr‐specific antibodies, within 3 months of seroconversion, appeared to be lower in the five men progressing to AIDS, compared with the men remaining symptom‐free during follow‐up. Analysis of the prevalence of previously described antibody profiles to these accessory gene products was carried out. In all HIV‐1 antibody seroconverters and in those HIV‐1 antibody seropositive men with 15 or more months of follow‐up who progressed to AIDS, there was a shift from predominantly nef‐ and vpu‐specific antibody negative profiles in the men developing AIDS in the early years of the study to predominantly nef‐ and vpu‐specific antibody positive profiles in men who developed AIDS later. Rev‐ and tat‐specific antibody negative profiles were dominant in men progressing to AIDS throughout follow‐up. No vpr‐specific antibody profile occurred preferentially in the men progressing to AIDS throughout follow‐up. Low antibody reactivity to accessory gene products nef, rev, and tat appears, like low anti‐core antibody reactivity, to be associated with progression to AIDS relatively rapidly after infection with HIV‐1.
Reduced duck hepatitis B virus viraemia in ducklings coinfected with the immunodepressive reticuloendotheliosis virus
Authors: Dixon Robert J. , Jones Noel F. , Freiman John S.
Published: March 2007

Coinfection of avian hosts by duck hepatitis B virus (DHBV) and reticuloendotheliosis virus (REV) was studied to assess the effect of immunodepression by REV on the replication of DHBV. One‐day‐old ducklings, domestic chickens, and turkey poults were inoculated either with DHBV or DHBV and REV and were bled and weighed at regular intervals. DHBV infection as manifested by viraemia and DHBV DNA in liver was established only in ducklings. All chickens and turkeys were negative for DHBV DNA in serum and liver. However, ducklings coinfected with REV showed a delayed onset and reduced level of viraemia compared to ducklings infected only with DHBV. The narrow host range of DHBV was confirmed even in immunodepressed species. It is suggested that the reduction in DHBV viraemia in ducklings was due to factors not involving the specific immune system.
Intrahepatic expression of HBcAg and delta antigen in anti‐HBe positive HBsAg carriers with acute exacerbation or chronic active liver disease
Authors: Chu Chia‐Ming , Liaw Yun‐Fan , Sheen I‐Shyan , Chen Tong‐Jong
Published: March 2007

Intrahepatic expression of HBcAg and hepatitis delta antigen (HDAg) was studied in 43 patients with acute exacerbation AE and 37 with chronic active liver disease CALD, in order to evaluate the role of hepatitis B virus (HBV) replication and hepatitis delta virus (HDV) superinfection in development of AE or CALD in anti‐HBe positive HBsAg carriers in Taiwan, and the results were compared with 37 patients with only minor hepatitic activity. Only 8.1% of patients with minor hepatitic activity were HBcAg positive and none were HDAg positive. In contrast, 41.8% and 32.6% of patients with AE were positive for HBcAg and HDAg, respectively, and the other 25.6% were negative for both. The clinical features of AE showed no difference in relation to HBcAg and HDAg activity in the liver except that patients without HBcAg and HDAg were predominantly older adults. These patients might be supposed to have non‐A, non‐B hepatitis (NANB) virus superinfection, as many reports have shown a predominance of older patients in acute NANB hepatitis. Of the patients with CALD, 40.5% were HBcAg positive, 27.0% were HDAg positive, and 32.5% were positive for neither. Histological features of CALD were identified in 83.3% and 100% of patients with HBcAg and HDAg activity, respectively, but only in 26.1% of those without HBcAg and HDAg. The etiology of CALD in the latter group remains unclear. Furthermore, some of the anti‐delta seropositive patients with AE or CALD expressed HBcAg rather than HDAg in the liver. In conclusion, the majority of AE or CALD instances in anti‐HBe carriers in Taiwan could be attributed to HBV replication or HDV superinfection. Studies of HBcAg and HDAg activity in the liver are thus of great importance in the diagnosis and management of HBsAg/anti‐HBe positive chronic liver disease.
Very low dose hepatitis B vaccination in the newborn: Anamnestic response to booster at four years
Authors: Moyes C. D. , Milne A. , Waldon J.
Published: March 2007

Seventy‐eight children who had received three very low doses (1 or 2 μg) of Merck, Sharp, and Dohme (MSD) plasma‐derived vaccine (PDV) in early infancy were followed to approximately four years of age. Of the 70 who had responded to the initial course of vaccine with measurable anti‐HBs, levels had fallen to below 10 mlU/ml in 38% of subjects given 1 μg doses and in 17% of those who had been given 2 μg doses. None of the children were positive for anti‐HBc. Two weeks after a 2 μg dose of MSD recombinant DNA (rDNA) vaccine all subjects had more than 10 mlU/ml of anti‐HBs, with 90% exceeding 1,000 mlU/ml. A response to hepatitis B vaccine in infancy is followed by an effective immunological memory for serveral years, even if anti‐HBs falls to low levels. The rDNA hepatitis B vaccine (MSD) in 2 μg doses is an effective booster following a primary course of plasma derived vaccine.
Serum antibodies to hepatitis C virus in Italian patients with hepatocellular carcinoma
Authors: Sbolli G. , Zanetti A. R. , Tanzi E. , Cavanna L. , Civardi G. , Fornari F. , Di Stasi M. , Buscarini L.
Published: March 2007

Antibodies against hepatitis C virus (anti‐HCV) were detected in 60.8% of 78 patients with hepatocellular carcinoma (HCC). Cirrhosis, present in most of the patients, as well as alcohol abuse, age, sex, and α‐fetoprotein were equally distributed in the anti‐HCV‐positive and ‐negative groups. HBsAg positivity was significatively higher in negative anti‐HCV group. By contrast, hepatitis B virus (HBV) antibodies were detected more frequently in positive anti‐HCV patients than in the negative anti‐HCV group. These data must be considered with caution becuase of the small number of HBsAg‐positive patients. It is concluded that the high prevalence of anti‐HCV in patients with HCC may suggest an etiological role of the hepatitis C virus, although in relationship to age, alcohol abuse and cirrhosis, the similarity in the two groups questions this hypothesis.
Detection of integrated hepatitis B virus DNA in hepatocellular carcinoma cell lines by nonradioactive in situ hybridization
Authors: Tay Nicholas , Chan Soh‐Ha , Ren Ee‐Chee
Published: March 2007

A sensitive and specific nonradioactive in situ hybridization method capable of detecting single‐copy integrated hepatitis B virus (HBV) DNA sequences in hepatocellular carcinoma (HCC) cell lines was developed. In situ hybridization of biotinylated HBV (adr, adw) DNA probes with nine different human HCC cell lines were carried out in 96‐well microtiter plates. Integration was detected in HCC cell lines HCCM, Hep3B, huH‐1, huH‐4, and PLC/PRF/5. Detection of single‐copy HBV DNA sequences was also achieved in Hep3B and huH‐4. HCC cell lines HepG2, HUH‐6, HUH‐7, Mahlavu, and the non‐HCC control MCF‐7, gave clear negative results. These results show a 100% correlation with those obtained by Southern blot hybridization assay. The results demonstrate that nonradioactive detection of single‐copy integrated HBV DNA sequences in HCC cell lines is possible by the method described, which has potential application for viral genome analysis requiring in situ hybridization.
Antibody‐ and complement‐mediated lysis of HIV‐infected cells and inhibition of viral replication
Authors: Gregersen J. P. , Mehdi S. , Baur A. , Hilfenhaus J.
Published: March 2007

HIV‐1‐positive antisera were tested for their ability to lyse HIV‐1‐infected cells in the presence of active complement. Cytolytic effects caused by sera derived from infected humans were slower than those observed with sera from immunised chimpanzees. Lytic but also negative sera were found among HIV‐1‐infected asymptomatic men as well as among clinical AIDS cases. Human antisera that lysed infected cells reacted similarly irrespective of whether the complement was heterologous or autologous. Analysis of complement‐mediated lysis using defined antisera against recombinant HIV‐1 env or core antigens suggested that gp 160/gp 120 and p24 can act as target antigens for an antibody‐ and complement‐mediated cytolysis of infected cells. Complement alone reduced the spread of HIV‐1 infection in CD4+ cells and the ability of HIV‐1 and HIV‐2 to form plaques in CD4‐transfected HeLa cells. Co‐operative effects of specific antibodies and complement were the most effective in inhibiting HIV infections.
Molecular epidemiology of adenoviruses isolated from hospitalized children with severe lower acute respiratory infection in Santiago, Chile
Authors: Kajon Adriana E. , Suarez Manuela Vicente
Published: March 2007

Genome analysis was performed on 69 adenovirus isolates from pharyngeal secretions of young children hospitalized with severe lower acute respiratory disease in Santiago, Chile, between 1984 and 1986. As expected, most isolated strains belonged to subgenus B (68.1%). Among the isolates of 1984, Ad7c was the dominant genotype (12 out of 23). The majority of isolates of 1986 were of the recently described genomic variant 3f.
Human papillomavirus infection of the uterine cervix analyzed by nonisotopic in situ hybridization
Authors: Hara Yoshihiro , Tuchida Shigeru , Nakamura Tetsuo , Yamamoto Kumio , Yamagata Shigefusa , Sugawa Tadashi , Minekawa Yoshiichi
Published: March 2007

Some types of human papillomaviruses (HPVs) have been suggested to be strongly related to uterine cervical carcinoma. An attempt to detect these in formalin‐fixed, paraffin‐embedded sections was made by either immunohistochemical or by in situ hybridization. Anticapsid protein of bovine papillomavirus antibody labeled with peroxidase was used for immunohistochemistry, and biotin was used instead of radioisotopes to label probes for in situ hybridization, which resulted in low background and a rapid procedure. Condylomatous changes were stained immunochemically with this antibody even in invasive carcinoma, whereas the carcinoma itself was not stained. Direct correlation was demonstrated by in situ hybridization between the HPV genome and histopathological structure, which was impossible by Southern or dot hybridization. HPV DNAs were detected in the nuclei of koilocytes and dyskeratinocytes of condylomata and dysplasias. Furthermore, hybridization signals of HPV DNAs in basal and parabasal cells suggested that HPV infection had already begun in the basal cells. In the case of malignant neoplasia accompanied by dysplasia, the same type of HPV was detected both in the malignant neoplasia and accompanying dysplasia. In one case of intraepithelial carcinoma, the very small focus of carcinoma just arisen in the cells of dysplasia was identified, and both were positive for HPV 18. This fact supports the suggestion that the carcinoma arises in dysplasia. Invasive carcinomas were classified further into keratinized, large‐cell nonkeratinized, and small‐cell nonkeratinized types, and the positive frequency for HPV 16 decreased as the differentiation of the carcinoma decreased. In the case of keratinized type of invasive carcinoma, strong hybridization signals were prominent around the malignant pearl formation.
Antiviral effect of antileukemic drugs N4‐Behenoyl‐1‐β‐D‐Arabinofuranosylcytosine (BH‐AC) and 2,2′‐Anhydro‐1‐β‐D‐Arabinofuranosylcytosine (Cyclo‐C) Against Human Cytomegalovirus
Authors: Nakamura Kazuo , Eizuru Yoshito , Kumura Keiko , Minamishima Yoichi
Published: March 2007

The antiviral activities of antileukemic drugs 1‐β‐D‐arabinofuranosylcytosine (Cytarabine; Ara‐C), 2,2′‐anhydro‐1‐β‐D‐arabinofuranosylcytosine (Ancitabine; Cyclo‐C), and N4‐behenoyl‐1‐β‐D‐arabinofuranosylcytosine (Enocitabine; BH‐AC) were evaluated in vitro against human cytomegalovirus (HCMV) in comparison with those of five other antiviral drugs. Both Ara‐C and Cyclo‐C showed the strongest inhibitory effect to HCMV. BH‐AC inhibited the replication of HCMV and depicted almost as the same dose‐response curve as Ganciclovir (DHPG). In the presence of Ara‐C, Cyclo‐C, or BH‐AC, triphosphate forms of the nucleoside analogs were detected in the HCMV‐infected cells, and synthesis of HCMV DNA was strongly suppressed. Thus, Ara‐C, Cyclo‐C, and BH‐AC were not only antileukemic, but also antiviral in vitro. However, Ara‐C and Cyclo‐C may not be suitable as anti‐HCMV agents, because they are cytotoxic or excreted rapidly in the urine in vivo [Van Voris, 1984; Hirayama et al., 1974]. Because of lower toxicity and longer retention in vivo, BH‐AC may be expected as an anti‐HCMV agent in patients with leukemia, in addition to serving as an antileukemic drug.
Association of antibodies blocking HIV‐1 gp160‐sCD4 attachment with virus neutralizing activity in human sera
Authors: Back Nicole K. T. , Thiriart Clotilde , Delers Anne , Ramautarsing Chitra , Bruck Claudine , Goudsmit Jaap
Published: March 2007

Sera, from HIV‐1 and HIV‐2 seropositive individuals, were tested for the presence of antibodies able to inhibit the binding (BI) of HIV‐IIIB gp160 (produced in mammalian cells using a vaccinia expression system) to the extracellular portion of the CD4 receptor. A competition enzyme immunoassay (EIA) with scluble CD4 (sCD4) was used. BI antibodies were highly prevalent among HIV‐1 seropositives but not in HIV‐2 infected individuals. Attempts to localize the binding site for these BI antibodies on the primary sequence of gp120 by using synthetic peptides encompassing the putative CD4 binding site on gp120 (aa 397–439) were not successful. This study did not reveal a significant correlation between the presence of BI antibodies and disease evolution. BI antibody titres correlated less well with anti‐gp160 titres (r=0.51, P⩽0.011) than with neutralizing antibody (NA) titres using either the isolates HIV‐SF2 (SF2) (r=0.77, P⩽0.000) and HIV‐MN (MN) (r=0.61, P⩽0.002) or the isolate HIV‐IIIB (HX10) (r=0.89, P⩽0.000) of which the gp160 for the assays was derived. An HIV‐IIIB neutralizing serum, elicited in a rabbit by immunization with a 17‐mer synthetic peptide derived from the third variable domain (V3) of gp120, did bind gp160 without inhibiting the subsequent attachment of sCD4 to gp160. These results indicate that (conformational) epitopes on gp160 recognized by antibodies that inhibit the attachment to CD4 may be identical or closely linked to the binding sites (not in the V3 domain) for HIV‐1 (group‐specific) neutralizing antibodies.
Time trends in the prevalence of human papillomavirus infections in archival Papanicolaou smears: Analysis by cytology, DNA hybridization, and polymerase chain reaction
Authors: Rakoczy Piroska , Sterrett Gregory , Kulski Jerzy , Whitaker Darrel , Hutchinson Laura , Mackenzie John , Pixley Ellis
Published: March 2007

A retrospective study was undertaken to determine the prevalence of human papillomavirus (HPV) infections in routine Papanicolaou (Pap) smears collected by general practitioners from Western Australian women in each of the years 1972, 1982, and 1987. HPV infection was detected by cytology, dot‐blot hybridization, or polymerase chain reaction (PCR). It was found that the prevalence of HPV infection remained unchanged over the 15 year study period, was independent of age, and was associated with normal cytology at a rate far greater than previously recognized. Indeed, the prevalence of cervical intraepithelial neoplasia (CIN) lesions, as detected by cytology, was 3.0% in 1972 and 3.8% in 1982 and 1987. The prevalence of HPV infection, detected as koilocytosis or parakeratosis, was 6.5%, 6.8%, and 5.3% in smears collected in 1972, 1982, and 1987, respectively, from 1,800 women. In 237 cytologically normal smears reprocessed for HPV‐DNA studies, the prevalence of HPV 16 was determined to be 15.6%, 11.2%, and 17.8% in 1972, 1982, and 1987, respectively, as determined by dot‐blot hybridization. However, the PCR detected HPV 16 in an additional 55.5%, 62.9%, and 57.0% of cytologically normal and dot‐blot negative smears. The prevalence of HPV 16 infection in cytologically normal smears was estimated to be 71.0%, 74.1%, and 74.8% in 1972, 1982, and 1987, respectively, by combining the HPV 16 dot‐blot and PCR‐positive results.

The high prevalence of HPV 16 in cytologically normal Pap smears suggests that infection with HPV 16, as detected by PCR amplification, does not place women in a high‐risk category for cervical cancer. In addition, we have shown that the application of PCR on reprocessed Pap smears is a powerful and sensitive method for retrospectively evaluating any aetiological role of HPV infection in the development of cervical cancer.
Differential cellular susceptibility to Epstein‐Barr virus infection in a patient with X‐linked lymphoproliferative disease
Authors: Okano Motohiko , Thiele Geoffrey M. , Gross Thomas G. , Davis Jack R. , Purtilo David T.
Published: March 2007

Epstein‐Barr virus (EBV) is often associated with lethal lymphoproliferative disease in immunologically compromised individuals. Recently, we have studied a 20‐month‐old boy with X‐linked lymphoproliferative disease (XLP) who had succumbed to infectious mononucleosis (IM) complicated by fulminant hepatitis and virus‐associated hemophagocytic syndrome following EBV infection. EBV genomes were detected in peripheral blood lymphocytes (PBL), cervical and mesenteric lymph nodes, liver, spleen, thymus, and bone marrow. According to restriction endonuclease analyses, the EBV‐DNA pattern was similar in all samples except for the EBV‐DNA from the bone marrow. Additionally, circular EBV‐DNA (suggesting a latent infection) predominated in spontaneously established lymphoblastoid cell lines (LCLs) derived from both the lymph node and cord lymphocytes co‐cultured with PBL. In contrast, both circular and linear EBV‐DNA (suggesting a lytic infection) were noted in spontaneously established LCLs derived from his PBL. Furthermore, LCLs derived from both the lymph node and cord lymphocytes co‐cultured with PBL expressed fewer reactive cells for early antigen (EA) and viral capsid antigen (VCA) than spontaneous LCLs from his PBL, thus providing evidence for different B cellular susceptibility to EBV infection in this patient with XLP. Finally, defective EBV‐specific cytotoxic T cell activity was observed in this patient. Latent EBV infected cells may easily escape immuno‐surveillance by the host. These findings may explain the fatal course of EBV infection in this patient.
Adenovirus infection in patients with Kawasaki disease
Authors: Okano Motohiko , Thiele Geoffrey M. , Sakiyama Yukio , Matsumoto Shuzo , Purtilo David T.
Published: March 2007

Two outbreaks of Kawasaki disease at different times and areas (Kyoto in 1982 and Sapporo in 1985) of Japan were studied retrospectively for the presence of antibodies to adenoviruses and herpesviruses. Only 2 of 12 (16.7%) consecutive acute phase sera of patients from the outbreak in 1982 and 1 of 10 (10.0%) sera from 1985 showed positive antibodies for the common adenovirus antigen by a complement fixation (CF) test, whereas 10 of 16 (62.5%) age‐ and sex‐matched controls during the outbreak of Kawasaki disease in 1985 were seropositive by the CF test. In contrast, using a recently developed enzyme‐linked immunosorbent assay (ELISA), 9 patients (75.0%) in 1982 and 9 patients (90.0%) in 1985 had antibodies to adenovirus type 2. In addition, 5 of 10 (50.0%) of the 1982 and 6 of 9 (66.7%) of the 1985 patients who were seronegative for CF antibodies were positive for IgM antibodies to adenovirus type 2. Fifteen (93.8%) controls were positive for antibodies to adenovirus type 2 by ELISA and only two sera showed negative CF antibodies with positive IgM antibodies to adenovirus type 2. Sequential sera from 4 patients in 1985 had either IgM or IgG antibodies by ELISA and eventually three became seropositive by the CF test in time. Additionally, no significant difference was noted with antibody status to herpes simplex virus type 1 and 2 (HSV‐1 and HSV‐2), varicella zoster virus (VZV), and cytomegalovirus (CMV) between patients and controls. In contrast, antibodies to Epstein‐Barr virus (EBV) were found in only one of 22 (4.5%) patients, whereas 13 (81.3%) controls were seropositive for EBV. These findings suggests that the primary infection with adenovirus type 2 may be associated with Kawasaki disease in patients who are seronegative for EBV.
Detection of human herpesvirus 6 DNA in throat swabs by polymerase chain reaction
Authors: Kido Shinji , Kondo Kazuhiro , Kondo Toshio , Morishima Tsuneo , Takahashi Michiaki , Yamanishi Koichi
Published: March 2007

The detection of human herpesvirus 6 (HHV‐6) DNA was carried out in throat swabs of adults and children by the polymerase chain reaction, and isolation of virus was also attempted from peripheral mononuclear cells. Although virus was isolated only from peripheral blood mononuclear cells of infants with exanthem subitum, HHV‐6 DNA was detected in one of 30 healthy adults (3%), two of nine adults (22%) with common cold, two of 10 infants (20%) with exanthem subitum, four of 39 febrile children (10%) with antibody to HHV‐6 (including three of 10 infants aged under 1 year old, and one of 29 children aged over 1 year old). However, HHV‐6 DNA was not detected in samples from healthy neonates.
Differentiation of primary cytomegalovirus infection from reactivation using the urea denaturation test for measuring antibody avidity
Authors: Blackburn N. K. , Besselaar T. G. , Schoub B. D. , O'Connell K. F.
Published: March 2007

Cytomegalovirus (CMV) is probably the most common agent of prenatal infection of the new‐born, and one of 20 congenitally infected newborns shows serious symptoms. It was therefore considered important to be able to differentiate primary CMV from reactivation in pregnant females. A urea denaturation test was used to distinguish primary from secondary rubella infection in which the urea is included in the wash step of the standard IgG ELISA. This resulted in the removal of low‐avidity antibodies, which are the antibodies produced early in infection. A group of CMV IgM‐negative and ‐positive sera were tested, and all but one showed moderate to high avidity, with an avidity index reading of more than 30%. Among a group of babies 3–12 months of age, who were CMV IgM positive, 55% (16 of 29) showed low‐avidity CMV antibodies. A small group of renal transplant patients and patients with clinically and laboratory‐confirmed CMV gave more or less predicted avidity index results. It appears that, with the method used at this laboratory, the urea denaturation test can be applied to CMV to determine primary infection or reactivation in the majority of cases.