bluuebunny/crossref_metadata_2025 huggingface dataset:
Protective antibodies to hepatitis B virus in haemophiliacs
Authors: Oon Lynette L. E. , King Alistair , Higgins Julie A. , Lee Christine A. , Kernoff Peter B. A. , Goodall Alison H.
Published: March 2007

Haemophilic patients are at increased risk from hepatitis B virus infection because of their need for blood product therapy. They are potentially poor responders to hepatitis B vaccine due immunological abnormalities resulting from two causes: infection with the human immunodeficiency virus and treatment with clotting factor concentrates. The protective antibody response to hepatitis B virus in vaccinated haemophiliacs was investigated using a competitive enzyme‐linked immunosorbent assay which employs monoclonal antibody, RF‐HBs‐1, that recognizes a virus‐neutralising epitope on HBsAg. Serum samples from 55 haemophilic patients were studied a t 7, 12, and 24 months after the first injection with HB vaccine. Twenty‐four vaccinated normal subjects were used as controls.

The level of neutralising antibody was found to correlate with the polyclonal anti‐HBs response in the majority of subjects in both the control and patient groups. There was a small but statistically significant reduction in both antibody responses in the patients compared with the normal controls. Treatment with FVIII or FIX concentrate did not influence the antibody response in the patients. Eleven of the haemophilic patients were anti‐HIV seropositive. This group had a significantly lower antibody response than anti‐HIV negative patients, and this correlated with the duration of anti‐HIV seropositivity, rather than with their T4 counts.

We conclude that, following vaccination, the majority of haemophiliacs are able to mount a protective antibody response to hepatitis B virus. HIV infection was found to be the sole cause of immunological suppression of this response.
Human papillomavirus infection of the male diagnosed by Southern‐blot hybridization and polymerase chain reaction: Comparison between urethra samples and penile biopsy samples
Authors: Kataoka Akio , Claesson Urban , Hansson Bengt Göran , Eriksson Margareta , Lindh Elisabeth
Published: March 2007

A group of 108 healthy men aged 18 to 23 years underwent penoscopy after application of acetic acid. In 39 (37%) aceto‐white lesions were found and a biopsy was taken. From 105 patients a urethra sample was collected with a brush. The prevalence of HPV‐DNA by dot‐blot and Southern‐blot hybridizations (DBH/SBH) and poly‐merase chain reaction (PCR) as well as the correlation between results from urethra samples and penile aceto‐white epithelium were examined. In urethra, HPV‐DNA was detected in four of the 66 patients (6%) with normal epithelium and in three of the 39 (8%) with aceto‐white epithelium by using DBH/SBH. By PCR the corresponding findings were eight (12%) and 10 (26%) HPV‐DNA‐positive patients, respectively. Out of the 39 biopsy samples HPV‐DNA was detected in six (15%) by DBH/SBH and in 17 (44%) by PCR. Twenty‐two (56%) of the patients with aceto‐white lesions were by PCR positive in either the urethra sample or biopsy. All samples positive by DBH/SBH were also positive by PCR except for one biopsy with HPV 35, a type not tested for by PCR. HPV 11 was the most common finding in urethra and HPV 6 and 16 in aceto‐white epithelium.
Detection of flaviviruses by reverse‐transcriptase polymerase chain reaction
Authors: Eldadah Zayd A. , Asher David M. , Godec Mark S. , Pomeroy Kitty L. , Goldfarb Lev G. , Feinstone Stephen M. , Levitan Herbert , Gibbs C. J. , Gajdusek D. Carleton
Published: March 2007

RNA sequences of five flaviviruses were detected by a modified polymerase chain reaction (PCR) that incorporated a reverse transcriptase and RNase inhibitor. Oligonucleotide primer pairs were synthesized to amplify sequences from St. Louis encephalitis (SLE), Japanese encephalitis (JBE), yellow fever (YF), dengue 2 (DEN‐2), and dengue 4 (DEN‐4) viruses. The amplified products were visualized as bands of appropriate size on ethidium bromide‐stained agarose gels. The identity of these products was confirmed by restriction endonuclease cleavage to generate fragments of predicted lengths. The reverse transcriptase PCR (RT‐PCR) successfully amplified flavivirus sequences from cell cultures, frozen brain tissue, and formalin‐fixed, paraffin‐embedded brain tissue. The reactions were highly specific, and the method compared favorably to two conventional assays of viral infectivity. RT‐PCR followed by PCR with nesting primers (N‐PCR) was 1,000‐fold more sensitive in detecting virus than classical infectivity titration by intracerebral inoculation of suckling mice and nearly 1,000‐fold more sensitive than amplification of virus in cell culture followed by inoculation of mice.
Human papillomavirus in exophytic condylomatous lesions on different female genital regions
Authors: Sugase Motoyasu , Moriyama Shin‐Ichi , Matsukura Toshihiko
Published: March 2007

Clinically diagnosed exophytic condylomatous lesions on the vulva (20 cases), vagina (5 cases), and cervix (9 cases) were examined pathologically, and human papillomavirus (HPV) types present in those lesions were identified by Southern blot hybridization analysis. All vulvar and vaginal lesions showed typical histopathological features of classical condylomata, and HPV 6 and 11 were found in 15 vulvar and 3 vaginal lesions and in 5 vulvar and 2 vaginal lesions, respectively. In 5 cervical lesions with typical condylomatous changes, HPV 6 or 11 was also detected; however, HPV 16 was found in 2 cases of cervical lesion surrounded by prominent intraepithelial neoplasia, and HPV 31 was found in 2 cases of slightly elevated lesion with intraepithelial neoplasia. These observations suggest that HPV 6 and 11 have the potency to induce the specific pathological changes, condylomatous, in any regions of the female lower genital tract.
Lack of immunity to hepatitis B virus among the expatriates residing in southern Taiwan
Authors: Chen Mei , Sun Cindy , Chen Wei‐Jen
Published: March 2007

Taiwan is an endemic area for hepatitis B virus (HBV) infection. There are more than 3‐million carriers on this island. Chronic HBV infections may result in liver cirrhosis and hepatocellular carcinoma, which are common in Taiwan. However little information is available concerning the immunity and the susceptibility to HBV among the expatriate‐population (non‐permanent residents from foreign countries) in Taiwan. Thus controversies exist on the indications for HBV vaccination among the foreign visitors and expatriates. This study evaluated the serological status of those who attended the Kaohsiung Adventist Clinic between May 1986 and August 1989. Hepatitis B surface antigen (HBsAg) and antibody (anti‐HBs) were determined by ELISA. A total of 152 expatriates (85M, 67F, aged 5 to 64 years, duration of residence in Taiwan: 2 weeks to 11 years) without previous HBV vaccination, were tested. HBsAg and anti‐HBs were not detected in 125 (82%). Anti‐HBs was found in 22 (15%). There was no correlation between the duration of residence and anti‐HBs positivity. However, there was an increased prevalence of anti‐HBs with increasing age after 30. For those expatriates who were anti‐HBs positive, 64% had a history of sexual contact with a partner from an endemic area of HBV. HBsAg was detected in 5 (3%). Three were symptomatic with abnormal liver function tests. Four had a history of sexual exposure. Their children were not antigenemic.

It is concluded that most (>82%) of the expatriates residing in southern Taiwan have no immunity to HBV and sexual contact is a major route (70%) of transmission. Vaccination is therefore strongly recommended for those expatriates who may have intimate relationships with local residents.
Hepatitis C virus antibody in hepatocellular carcinoma in Taiwan
Authors: Jeng Jen‐Eing , Tsai Jung‐Fa
Published: March 2007

The prevalence of antibody to hepatitis C virus (anti‐HCV) was investigated in patients with hepatocellular carcinoma (HCC), and correlated with the clinical features. Anti‐HCV was detected in 129 histology or aspiration cytology proven HCC patients and 54 healthy controls. Anti‐HCV was examined by the HCV EIA (Abbott Laboratories). All healthy controls were anti‐HCV‐negative. Nineteen of 81 (23.5%) hepatitis B surface antigen (HBsAg)‐positive HCC patients were positive for anti‐HCV. Anti‐HCV was found among 60.4% (29/48) of HCC patients without detectable HBsAg. Forty‐eight of 129 (37.2%) HCC patients were positive for anti‐HCV. There was a significant difference in the prevalence of anti‐HCV between patients with HBsAg (23.5%) and those without HBsAg (60.4%, P = 0.0001). However, irrespective of the status of HBsAg, there was no statistical difference in sex, age, routine liver function tests, alpha‐fetoprotein concentration, or associated cirrhosis between patients with anti‐HCV and those without. The results imply that hepatitis C virus may play a role in the pathogenesis of HCC.
Seroprevalence of hepatitis B virus infection in children in Taipei, 1989: Five years after a mass hepatitis B vaccination program
Authors: Tsen Yee‐Jeng , Chang Mei‐Hwei , Hsu Hong‐Yuan , Lee Chin‐Yun , Sung Juei‐Low , Chen Ding‐Shinn
Published: March 2007

A nationwide hepatitis B vaccination program was launched in Taiwan in 1984. To study the impact of this ongoing program on hepatitis B virus (HBV) infection, a follow‐up seroepidemiologic study was carried out in 1989 in a Taipei district where pre‐vaccination seroepidemiology had been studied. HBV markers were studied in 1134 apparently healthy children (619 boys and 515 girls) under 13 years of age between March and July 1989.

The prevalence of hepatitis B surface antigen (HBsAg) in children under 5 years of age decreased from 9.3% in 1984 to approximately 2% in 1989. A significant decrrease in HBsAg prevalence and hepatitis B core antibody in 5‐ to 8‐year‐old children who were not immunized against HBV showed that horizontal infection among the older children had also decreased. Thus, this program not only protected vaccinated subjects; the reduction in numbers of highly infectious young HBV carriers also contributed to a lower prevalence of hepatitis B infection and carrier rates in some older children. This study demonstrates that hepatitis B vaccination is effective in protecting the majority of children in hyperendemic areas from HBV infection and from becoming chronic carriers.
Characterisation of a series of human immunodeficiency virus isolates derived sequentially from a single patient
Authors: Farrar Graham H. , Roff Marilynn A. , Amin Tehmina , Ball Jonathan , Parrett Alison M. , Battacharyya Uma , Booth James , Wansbrough‐Jones Mark H. , Greenaway Peter J.
Published: March 2007

Five HIV‐1 isolates were obtained sequentially from a single seropositive individual during the later stages of AIDS. Four of these isolates were adapted to grow in a continuous human T‐lymphocytic cell line. Comparative biological and biochemical studies of the virus isolates were made using persistently infected cultures or virus derived from these systems respectively. The data obtained clearly shows that viruses with different biological properties can be isolated from the same individual at different times during the course of clinical AIDS.
Chronic fatigue syndrome: Lack Of association with hepatitis C virus infection
Authors: Dale Janet K. , Di Bisceglie Adrian M. , Hoofnagle J. H. , Straus Stephen E.
Published: March 2007

Chronic fatigue syndrome (CFS) is a debilitating heterogeneous disorder lacking consistent, objective physical or laboratory abnormalities. Among the hypothetical etiologies for CFS are chronic viral infections. The present controlled seroprevalence study found that, among typical CFS patients, evidence of hepatitis C virus (HCV) infection is uncommon. Only one of 36 patients and none of 14 controls were anti‐HCV positive. The positive patient had persistent aminotransferase elevations and prior posttransfusion hepatitis. Thus HCV infection is not a common feature of CFS and should not be routinely sought.
Autoantibodies during α‐interferon therapy for chronic hepatitis B
Authors: Fattovich G. , Betterle C. , Brollo L. , Pedini B. , Giustina G. , Realdi G. , Alberti A. , Ruol A.
Published: March 2007

The development of autoantibodies and autoimmune reactions has been reported during and after interferon (IFN) therapy. Thirteen different antibodies from the sera of 32 patients with chronic hepatitis B treated with α‐interferon (α‐IFN) were tested. Seventeen HBeAg‐positive patients received 4.5 megaunits (MU) of recombinant IFN thrice weekly for 4 months, and 15 anti‐HBe and HBV‐DNA positive patients were treated with 5 MU/m2 of lymphoblastoid IFN thrice weekly for 6 months. Five patients (15%) had antinuclear antibodies (ANA) and one patient (3%) had smooth muscle antibodies before treatment. ANA appeared during IFN treatment in five (18%) of 28 previously negative patients. With discontinuation of treatment, the titer of ANA fell to undetectable levels in all patients. In contrast, none of the patients developed antibodies to endocrine organs, such as thyroid microsomal, thyroglobulin, parietal cells, pancreatic islet cell, and adrenal cortex antibodies or autoantibodies specifically associated with autoimmune liver disease such as liver kidney microsomal antibodies and antimitochondrial antibodies. There was no correlation between autoantibody positivity before therapy or autoantibody occurrence during treatment and response to IFN therapy. None of the patients developed clinical signs of autoimmune disease. These results indicate that these regimens of recombinant and lymphoblastoid IFN therapy of chronic hepatitis B are associated with a low risk of clinically significant autoimmunity.
Activation of cellular oncogenes by clinical isolates and laboratory strains of human cytomegalovirus
Authors: Boldogh I. , Abubakar S. , Fons M. P. , Deng C. Z. , Albrecht T.
Published: March 2007

The effect on cellular (c) oncogene RNA levels was investigate after infection of permissive cells with cell culture adapted strains (AD‐169, C‐87, Davis) and unadapted clinical isolates (82–1, 84–2, 85–1) of human cytomegalovirus (HCMV). The results indicate that both adapted and unadapted strains of HCMV induce substantial increases in c‐oncogene RNA levels for fos, jun, and myc measured by Northern blot hybridization. Elimination of immediate early (IE) protein synthesis between 0 and 3 hrs or reduction of virus infectivity (99.99%) by UV‐irradiation did not reduce the increase in c‐oncogene RNA levels. Inhibition of viral and cellular protein synthesis by cycloheximide resulted in a high abundance (superinduction) of specific RNAs which hybridized to c‐oncogene probes after infection with either adapted or unadapted strains of HCMV. These data suggest that IE viral gene expression is not essential for activation of c‐oncogenes. Inhibition of DNA‐dependent RNA synthesis by blocking RNA elongation with actinomycin‐D or by inhibiting the activity of RNA polymerase II with alpha‐amanitin significantly reduced the increase in c‐oncogene RNA levels, suggesting that activation of cellular genes by HCMV is controlled at the level of transcription. Activation of c‐oncogenes by HCMV may be particularly important because their protein products appear to be involved in initiation and regulation of viral and cellular gene expression.
Primary and recurrent cytomegalovirus infections have different effects on human herpesvirus‐6 antibodies in immunosuppressed organ graft recipients: Absence of virus cross‐reactivity and evidence for virus interaction
Authors: Ward K. N. , Sheldon M. J. , Gray J. J.
Published: March 2007

The relationship between serum antibodies to human herpesvirus‐6 (HHV‐6) and cytomegalovirus (CMV) infection was studied in immunosuppressed adult organ graft recipients all of whom had IgG to both HHV‐6 and Epstein‐Barr virus capsid antigen (EBVCA) before operation and who had received an organ or organs from HHV‐6 seropositive donors. In primary CMV infection the titre of IgG to HHV‐6 rose substantially (between 32‐ and 512‐fold) in eight out of eight patients whereas JgG to EBVCA only rose 32‐fold in two patients. Moreover, the HHV‐6 responses coincided closely with the CMV seroconversion. Serum absorption studies gave no evidence for anti body cross‐reaction between CMV and HHV‐6 because the CMV antibody titre could be reduced specifically without affecting HHV‐6 antibody titres and vice versa. In recurrent CMV infection, HHV‐6 antibody levels rose (32‐fold) in three out of eight patients but these changes did not coincide with the CMV antibody response. Similarly, in the complete absence of CMV infection, five out of eight patients showed antibody rises to HHV‐6 (between four‐ and 16‐fold). IgG titres to EBVCA were stable in both these groups of patients. It is concluded that there is serological evidence (rising titre ≥ four‐fold) for genuine HHV‐6 reactivation or, alternatively, for reinfection in 16 out of the 24 patients. This phenomenon was most frequent in primary CMV infection where the largest HHV‐6 antibody responses were seen probably because of an, as yet, undetermined interaction between the two viruses.
Is vestibular papillomatosis associated with human papillomavirus?
Authors: Costa Silvano , Rotola Antonella , Terzano Patrizia , Secchiero Paola , Luca Dario Di , Poggi Maria G. , Masotti Patrizia , Martinell Giuseppe , Cassai Enzo
Published: March 2007

The origin and clinical significance of vestibular papillae were evaluated by comparing histological features with the presence of human papillomavirus (HPV) types 6/11 and 16/18, as revealed by Southern blot DNA hybridization. Twenty women with vestibular papillomatosis underwent clinical evaluation and follow‐up. When available, male partners were also examined. Histological changes suggestive of HPV infection were present in all the 20 specimens. Sixteen cases (80%) contained DNA sequences homologous to the viral probes. In particular, 12 cases (60%) reacted with the HPV 16/18 probe. Follow‐up for more than 18 months revealed no variation in the distribution and appearance of vestibular papillae. No male partner showed signs of HPV lesions. The study shows that HPV 16 is frequently associated with vestibular papillae but does not support a productive infection. Therefore the most appropriate management of these patients should be evaluated clinically in each individual case.
Murine acinar pancreatitis preceding necrotizing myocarditis after coxsackievirus B3 inoculation
Authors: Gómez Ricardo M. , Lascano Eduardo F. , Berrìa Marìa I.
Published: March 2007

Balb/C weanling mice were inoculated intraperitoneally with a myocarditic variant of coxsackie virus B3, with the aim of characterizing more fully the cell damage induced in the heart as well as in other organs. During the first week postin‐fection (pi), all animals developed acinar pancreatitis, followed by focal myocarditis. In accordance with the increasing infectivity titers, such progressive histopathological changes correlated with local viral replication. From day 4 pi, acinar degeneration accompanied by diffuse inflammatory exudate was observed in the pancreas, followed by fatty tissue replacement by day 8. In the heart, focal necrosis rather than inflammatory reaction first appeared at 4 days pi and became widespread by 6–8 days pi. Necrotic foci usually presented calcium deposits, with absence of myofibrils in the affected fibers. The fact that both periodic acid Schiff (PAS) and Best carmine staining remained positive even after diastase treatment ruled out basophilic necrosis. In summary, the pancreas appeared to be the site of primary viral replication leading to viremia.
Rhinovirus induces natural killer‐like cytotoxic cells and interferon alpha in mononuclear leukocytes
Authors: Levandowski Roland A. , Horohov David W.
Published: March 2007

Natural killer‐like cellular cytotoxicity was augmented by incubation of human rhinovirus serotype 2 with peripheral blood mononuclear leukocytes collected from healthy donors. The production of alpha interferon but not gamma interferon was identified in the same cell cultures. A specific interaction of conformationally intact rhinovirus with peripheral blood mononuclear leukocytes was required for induction of the response, since the response was extinguished at reduced quantities of infectious rhinovirus, and acid inactivated rhinovirus did not augment cellular cytotoxicity. Productive replication of rhinovirus was not observed in cultures of peripheral blood mononuclear leukocytes. The replicative failure was not related merely to interferon production, since the rate of disappearance of rhinovirus was similar to that observed in cell free medium. The findings suggest that natural killer cells should be considered as a potential component of the local nasopharyngeal pathophysiology of rhinovirus infection.
Cytomegalovirus IgG and IgA serum antibodies in a study of HIV infection and HIV related diseases in homosexual Men
Authors: Levy Esther , Margalith Miriam , Sarov Batia , Sarov Israel , Rinaldo Charles R. , Detels Roger , Phair John , Kaslow Richard , Ginzburg Harold , Saah Alfred J.
Published: March 2007

The significance of serum IgG and IgA antibodies to cytomegalovirus (CMV) at various stages of human immune deficiency virus (HIV) infection was studied in 175 homosexual men. Sera were obtained from 123 HIV seropositives [41 asymptomatic, 29 with lymphadenopathy associated syndrome (LAS), 22 with AIDS related complex (ARC), and 31 AIDS patients], 17 HIV seroconverters, and 35 HIV asymptomatic seronegatives. The sera were tested blindly for CMV IgA and IgG antibodies using the immunoperoxidase assay (IPA) and CMV infected human embryo cells. Crossectional analysis of CMV IgG antibodies at a titer of ⩾20 showed 87% and 100% prevalence in the HIV seronegative groups and in the HIV seropositive groups, respectively (P < 0.05). CMV IgG antibodies at a titer of ⩾80 were present in significantly higher proportions among the HIV seropositive subjects of the various groups as compared with the HIV seroneg‐ ative homosexual men. However, in the HIV seronegatives who later seroconverted to HIV, a significantly higher prevalence of CMV antibodies (35%) was detected before HIV seroconversion, as compared with the persistently HIV seronegative subjects (14.3%) (P < 0.05). The HIV seronegatives pre‐HIV seroconversion also exhibited a significantly higher geometric mean titer (GMT) of CMV IgG antibodies (62.17 ± 0.64) as compared with the persistently HIV seronegatives (34.0 ± 0.6) (P = 0.03). Significantly higher GMTs of CMV IgG antibodies were detected in all the HIV seropositive groups as compared with the persistently HIV seronegativegroup.

CMV IgG antibodies were not detected in the HIV seronegative subjects. CMV IgA antibodies were found in 5.9% of the HIV seroconverters, i n 19.5% of the HIV seropositive asymptomatic subjects, 34.5% of LAS, 22.7% of ARC, and 6.5% of AIDS patients. These data support the concept that CMV might be associated with susceptibility of homosexual men to HIV infection and disease progression.
Generation of a 1.5‐kb cDNA fragment of the hepatitis C virus genome by overlap extension
Authors: Tong Shuping , Li Jisu , Vitvitski Ludmila , And Daniel Lepot, , Trépo Christian
Published: March 2007

Hepatitis C virus (HCV) is a recently identified RNA virus responsible for most of non‐A, non‐B hepatitis. Genetic analysis of HCV sequences and their encoded proteins has been hampered by the difficulty in obtaining cDNA fragments of sufficient lengths: construction of cDNA library requires technical expertise while amplification by the polymerase chain reaction (PCR) usually yields fragments of less than 400 base pairs. In this report we have generated a 1.5‐kb HCV cDNA fragment by overlap extension of smaller PCR fragments and by ligation through restriction sites. Sequencing of the cloned fragment confirmed the absence of significant sequence alteration produced by this procedure. Overlap extension may represent an easy method for generating relatively large HCV cDNA clones for the expression of HCV‐encoded proteins.
Changing seroepidemiological patterns of cytomegalovirus infection in children in Taiwan from 1984 to 1989
Authors: Lee Ping‐Ing , Chang Mei‐Hwei , Lee Chin‐Yun , Kao Chuan‐Lian
Published: March 2007

The prevalence of cytomegalovirus (CMV) antibody was studied in 966 children in 1984 and 927 children in 1989. The overall prevalence rate of CMV antibody was 59% for children in 1984 and 46% for children in 1989 (P < 0.05). In both study years, the prevalence rate of CMV antibody was about 70% in infants under 6 months of age, declined to a trough between the ages of 6 and 12 months, and then increased to 40‐50% between 1 and 4 years of age. The rate of CMV antibody for children above 4 years in 1984 increased steadily with age and reached 82% by 12 years. In contrast, the prevalence rate in 1989 remained at the level of 40‐5070 from age 4 to 10 years. It was followed by a sharp increase after 10 years of age and reached 84% at 12 years old. The seropositive rate in each of the 5‐, 6‐, 8‐, 9‐, and 10‐year‐old groups was higher in 1984 than that in 1989. These observations indicated that the prevalence rate of CMV antibody is decreasing in children. This change may be related to various socioeconomic factors, especially the less crowded family conditions in recent years.
Sequence analysis of an HIV‐1 isolate which displays unusually high‐level AZT resistance in vitro
Authors: Muckenthaler M. , Gunkel N. , Levantis P. , Broadhurst K. , Goh B. , Colvin B. , Forster G. , Jackson G. G. , Oxford J. S.
Published: March 2007

Multiple mutations in the reverse transcriptase (RT) gene were observed in a drug‐resistant isolate of human immunodeficiency virus type 1 (HIVI) from an individual having prolonged (<2 years) zidovudine (AZT) therapy. The virus replicated in PBMC′s in the presence of very high concentrations of AZT (1 25 μM). Drug‐sensitive strains were curtailed by 0.01 μM AZT.

Eleven defined mutations were observed as compared with published sequences of RT for eight strains of HIVI. Eight of these mutations were found in the domain involved in nucleotide recognition and enzyme function. Only one of the mutations, giving a Thr—Tyr change at amino acid 215, matched those previously ascribed (67,70, 215, and 219) to the generation of high‐level resistance to AZT. Therefore additional amino acid changes may have significance in the emergence of super‐resistant viruses.
Sequence variation of functional HTLV‐II tax alleles among isolates from an endemic population: Lack of evidence for oncogenic determinant in tax
Authors: Hjelle Brian , Chaney Rebecca
Published: March 2007

Human T‐cell leukemia‐lymphoma virus type II (HTLV‐II) has been isolated from patients with hairy cell leukemia (HCL). We previously described a population with longstanding endemic HTLV‐II infection, and showed that there is no increased risk for HCL in the affected groups. We thus have direct evidence that the endemic form(s) of HTLV‐II cause HCL infrequently, if at all. By comparison, there is reason to suspect that the viruses isolated from patients with HCL had an etiologic role in the disease in those patients. One way to reconcile these conflicting observations is to consider that isolates of HTLV‐II might differ in oncogenic potential. To deter mine whether the structure of the putative oncogenic determinant of HTLV‐11, tax2 might differ in the new isolates compared to the tax of the prototype HCL isolate, MO, four new functional tax cDNAs were cloned from new isolates. Sequence analysis showed only minor (0.9‐2.0%) amino acid variation compared to the published sequence of MO tax2 Some codons were consistently different from published sequences of the MO virus, but in most cases, such variations were also found in each of two tax2 clones we isolated from the MO T‐cell line. These variations rendered the new clones more similar to the tax1 of the pathogenic virus HTLV‐I. Thus we find no evidence that pathologic determinants of HTLV‐II can be assigned to the tax gene.
Prevalence of antibodies to recombinant virion infectivity factor in the sera of prospectively studied patients with HIV‐1 infection
Authors: Schwander S. , Braun R. W. , Kühn J. E. , Hufert F. T. , Kern P. , Dietrich M. , Wieland U.
Published: March 2007

Two hundred twenty‐four sera were collected from 34 HIV‐1 infected patients during an observation period of up to 4.5 years (109 patient years of observation). The sera were tested for the presence of antibodies against the HIV‐1 virion infectivity factor (vif) protein. Thirty sera from 6 HIV‐1 seronegative individuals served as negative controls. The sera were immunoblotted against a recombinant, prokaryotically expressed vif protein. The prevalence of anti‐vif antibodies increased significantly with progression of the disease from 18% to 81% (P < 0.0001) which suggests a possible role of vif in HIV‐1 replication and pathogenicity.
Human infections with Borna disease virus: Seroprevalence in patients with chronic diseases and healthy individuals
Authors: Bode L. , Riegel S. , Lange W. , Ludwig H.
Published: March 2007

Borna disease virus (BDV) is as yet an unclassified infectious agent which causes a neurologic disease in horses and sheep and is transmissible to other animal species. Human sera were tested for BDV‐specific antibodies by immunofluorescence and immunoprecipitation. The sera were collected from three thousand subjects in Europe, the USA, and Africa classified by clinical syndromes/infectious diseases, and from healthy controls. In all three continents, positive serum samples were detected which recognized the major BDV antigen of 38/40 KD present in the nuclei of infected cells. In contrast to 2% of se‐ropositives among the normal population, significantly higher prevalences (13‐14%) were present among patients with chronic progressive diseases of the brain and the immune system. In children, antibody frequencies were two to four times higher than in adults. The results suggest a latent infection with BDV in humans leading to low antigen expression in healthy subjects and frequent reactivation events in chronically ill patients.
Detection of HPV‐16 in cell lines and cervical lavage specimens by a polymerase chain reaction‐enzyme immunoassay assay
Authors: Coutlée François , Bobo Linda , Abbass Hawwari , Dalabetta Gina , Hook Ned E. , Shah Keerti , Viscidi Raphael P.
Published: March 2007

A gene amplification method that combines the polymerase chain reaction with detection of amplified DNA in a solution hybridization/enzyme immunoassay (PCR‐EIA) was developed for HPV‐16 DNA. Samples were amplified with primers for the E7‐E1 region of HPV‐16. Amplified DNA products were identified and quantitated by hybridization in solution with a biotinylated RNA probe. Labeled DNA/RNA hybrids were measured semiquantitatively in an enzyme immunoassay using solid phase anti‐biotin antibody and liquid phase B‐d‐galactosidase labeled monoclonal antibody against DNA‐RNA hybrids. Enzyme bound to the solid phase was quantitated with a fluorogenic substrate. The assay was linear over 2 log10 dilutions of SiHa cells and the detection limit was three copies of HPV‐16 genome. The sensitivity of PCR‐EIA for detection of PCR amplified products compared favorably with slot and Southern blots using a 32P‐labeled RNA probe. The assay was used to assess HPV‐16 infection of uterine cervix in women attending a clinic for sexually transmitted diseases. Twenty‐one of the 81 specimens (25.9%), obtained by cervicovaginal lavage, were positive for HPV‐16 by PCR‐EIA. The assay provides a convenient means to objectively measure HPV DNA amplified with PCR. © 1992 Wiley‐Liss, Inc.
Hepatitis E virus: The cause of a waterbourne hepatitis outbreak
Authors: Skidmore S. J. , Yarbough P. O. , Gabor K. A. , Reyes G. R.
Published: March 2007

Newly developed assays for antibody to hepatitis E virus (HEV) were used to study 114 serum samples collected during an outbreak of enterically transmitted hepatitis that occurred in Kashmir in 1978/9. The sera included samples from patients with viral hepatitis, anicteric hepatitis, contacts of cases, and unaffected persons. A total of 71% of patients with viral hepatitis were found positive for anti‐HEV specific IgG, and 75% of these were also positive for IgM. These data confirm the hepatitis E virus as the causative agent in this outbreak. © 1992 Wiley‐Liss, Inc.
Fine specificity of the humoral immune response to HIV‐1 GP160 in HIV‐1 infected individuals from tanzania
Authors: Nkya Watoky M. M. M. , Warren Ronald Q. , Wolf Hans , Hendrix Craig W. , Tesha John , Redfield Robert R. , Melcher Gregory P. , Burke Donald S. , Kanda Patrick , Kennedy Ronald C.
Published: March 2007

A total of 160 sera from HIV‐1 infected individuals from Tanzania were examined for their fine specificity characteristics relative to 9 synthetic peptides that define HIV‐1 gp160 epitopes. Immunorecessive and immunodominant epitopes were identified in both gp120 and gp41 based on serologic reactivity of these HIV‐1 infected sera. A significant difference in fine specificity among HIV‐1 infected individuals from Tanzania and the United States was observed for an immunodominant gp41 epitope. No significant differences in reactivity among asymptomatic vs. symptomatic HIV‐1 infected individuals were detected for the selected HIV‐1 gp160 epitopes defined by these peptides. The majority of sera from HIV‐1 infected Tanzanians contained antibodies that recognized a peptide corresponding to the V3 region of gp120 from the HIV‐1 MN isolate. These data suggest that regional isolates of HIV‐1 may exist in Tanzania that differ from HIV‐1 isolated in the United States. However, based on serology, HIV‐1 isolates exhibiting sequences with HIV‐1 MN V3 similarity may also be prevalent in Tanzania. The results of this study may be useful for the design of more effective AIDS diagnostic and therapeutic products for use worldwide. © 1992 Wiley‐Liss, Inc.
Inhibition of HIV‐1 infection in vitro by murine monoclonal anti‐p24 antibodies
Authors: Franke L. , Grunow R. , Meissner K. , Porstmann T. , von Baehr R.
Published: March 2007

Murine monoclonal antibodies (Mabs) to the major core protein p24 of the human immunodeficiency virus type 1 (HIV‐1) were tested for their ability to inhibit the replication and spread of the virus in permanent cell cultures (Molt4/8, K37, H9) and in the culture of 11‐2 stimulated T cells of healthy donors. After addition of ascitic fluid containing monoclonal anti‐p24 antibodies or purified anti‐p24 antibodies or the respective control to co‐cultures of infected and non‐infected cells, HIV‐1 replication was evaluated by determining the percentage of infected cells and the activity of reverse transcriptase (RT) in cell‐ free supernatant. In addition, the supernatant's infectivity was determined.

FACS analysis demonstrated p24 antigen in about 40% of unfixed HIV‐1 infected cells at the cell membrane. Monoclonal anti‐p24 antibodies of different epitope specificity added to the cells but not to the virus delayed the spread of HIV‐1 infection in permanent cell culture.

Furthermore, anti‐p24 Mabs inhibited the re‐ lease of RT‐active virus particles by HIV‐1 infected cell lines or 11‐2 stimulated T‐lymphocytes, respectively, up to 60%. The mode of action of anti‐p24 antibodies after HIV‐1 infection is discussed on the basis of the data obtained. © 1992 Wiley‐Liss, Inc.
The interrelationship between HBV‐markers and HIV antibodies in patients with hepatocellular carcinoma
Authors: Tswana S. A. , Moyo S. R.
Published: March 2007

To determine the interrelationship between hepatitis B viral markers (HBV), the human Immunodeficiency virus (HIV), and hepatocellular carcinoma (HCC) in HCC patients, a total of 282 subjects were included in the study. Out of 282 subjects, 182 were HCC patients as determined by raised α‐feto‐protein (AFP) of >1,000 ng/ml. The other 100 control patients presented with other conditions and had detectable AFP of <1,000 ng/ml i n their sera.

On presentation, 10 ml of venous blood was drawn from each enrolled subject and taken to the laboratory.

HBV markers were detected using commercial reagents; HIV antibodies were detected by the commercial ELISA tests and were confirmed by Western blot. AFP was detected using an RIA technique.

Of 282 examined subjects 182 (64.5%) had detectable AFP of >1,000 ng/ml. 113 (40.1%) and 103 (36.5%) had HBsAg and Anti‐HBc respectively. However, HBeAg was found in 21 of 113 (18.6%) of the HBsAg positive only. Anti‐HIV anti‐bodies were present in 15 (5.3%) of the 282 tested individuals. Only 1 (1.0%) of the control group had detectable anti‐HIV antibodies in the serum. Eleven percent and 4.0% of the same control group had HBsAg and anti‐HBc in their sera respectively.

The study shows a significant correlation between HCC and HBV‐markers (P < 0.0001). Similarly, a significant correlation between anti‐HIV antibodies and HBV‐markers, (P < 0.0001) was found. © 1992 Wiley‐Liss, Inc.
Detection of HCV RNA in saliva, urine, seminal fluid, and ascites
Authors: Liou Tai‐Cherng , Chang Ting‐Tsung , Young Kung‐Chia , Lin Xi‐Zhang , Lin Ching‐Yih , Wu Hua‐Lin
Published: March 2007

Approximately half of the patients with type C hepatitis do not have a history of parenteral exposure. The route of nonparenteral infection remains unknown. To evaluate the possible role of body fluids, the existence of hepatitis C virus (HCV) RNA in saliva, urine, seminal fluid, and ascites was examined by “nested” polymerase chain reaction (PCR). Amplification of the HCV 5′ noncoding sequences was carried out. The amplified product was confirmed by Southern blot hybridization and restriction endonuclease digestion. Among 34 patients with chronic liver disease who were positive for anti‐HCV and serum HCV RNA, the prevalence of HCV RNA in body fluids was 100% (7/7) in ascites, 48% (1 5/31 ) in saliva, 24% (4/17) in seminal fluid, and 7% (2/29) in urine. The body fluids collected from 3 healthy subjects and 5 patients with chronic liver disease who were positive for anti‐HCV but negative for serum HCV RNA were all negative for HCV RNA. Hence, the potential infectivity of body fluids in patients testing negative for serum HCV RNA can probably be discounted. Conversely, the presence of HCV RNA in saliva and seminal fluid of patients positive for serum HCV RNA suggests sexual and household contact as likely modes of nonparenteral transmission of type C hepatitis. Furthermore, the high prevalence of HCV RNA in ascites and saliva may have important implications in medical and dental practice. © 1992 Wilev‐Liss, Inc.
Patterns of serological markers in transfusion‐transmitted hepatitis C virus infection using second‐generation HCV assays
Authors: Lelie P. Nico , Cuypers H. Theo M. , Reesink Henk W. , van der Poel Cees L. , Winkel Irene , Bakker Ed , van Exel‐Oehlers Patricia J. , Vallari David , Allain Jean Pierre , Minims Larry
Published: March 2007

A semiautomated dot blot assay and cDNA polymerase chain reaction (PCR) were used to study longitudinal anti‐hepatitis C virus (HCV) recognition patterns i n relation to presence of HCV‐RNA in transfusion recipients and their infectious donors. In 9 recipients, 4 different patterns of HCV infection were observed: (A) persistent HCV carriage accompanied by chronic hepatitis in 6, (B) acute resolved hepatitis, but persistent HCV replication in one, and (C) continuous HCV replication without hepatitis in one and (D) acute resolved hepatitis with clearance of infection in one. This last self‐limited infection was characterized by the disappearance of HCV‐RNA as well as anti‐HCV reactivity. In contrast, antibody reactivity persisted in 7 of 8 patients with chronic HCV infection who could be followed until 1990. Seven of the 9 recipients developed antibodies to all recombinant peptides in dot blot assay; one became positive for anti‐C33 and anti‐core and one developed anti‐core only. The sequence of appearance of antibodies differed among individual patients. In 7 patients with full anti‐HCV recognition patterns, the sequence of events was (mean and limits in days after transfusion): onset of hepatitis at day 50 (22–74), seroconversion of anti‐C33 at day 91 (59–129), anti‐core at day 133 (54–203), and anti‐C100 at day 143 (59–365). The incorporation of C33 and core proteins, in addition to C100, in the second generation anti‐HCV ELISA enhanced the detection rate in the HCV‐infected transfusion recipients from 7/9 (78%) to 9/9 (100%). In 4 of the 7 anti‐C100 seroconverters anti‐HCV was detectable 38–277 days earlier in the new ELISA. However, the development of anti‐HCV in the second‐generation ELSA was still delayed [54–192 (mean 103) days after transfusion and 0–135 (mean 53) days after onset of hepatitis]. We conclude that combined application of anti‐HCV immunoblot techniques and cDNA‐PCR in consecutive samples is necessary for reliable diagnosis of either ongoing productive or resolving HCV infection. © 1992 Wiley‐Liss, Inc.
Increased expression of transforming growth factor α after transfection of a human hepatoblastoma cell line with the hepatitis B virus
Authors: Tabor Edward , Farshid Mahmood , Bisceglie Adrian Di , Hsia Chu Chieh
Published: March 2007

The expression of transforming growth factor a (TGF‐α) was examined in a human hepatoblastoma cell line. Hep G2, which does not contain hepatitis B virus (HBV) DNA, and in the cell line 2.2.15, which was formed by the transfection of Hep G2 cells with the complete HBV DNA, to study the possibility that HBV and TGF‐α could function as co‐factors in hepatocarcinogenesis. Northern blot hybridization of RNA extracted from these cell lines, with densitometric analysis, revealed expression of the TGF‐α gene in the transfected cells at a level three times higher than in the nontransfected cells. Staining of the cells using a monoclonal antibody to TGF‐α and the avidin‐biotin‐peroxidase immunohistochemical method revealed a much higher intensity of TGF‐α staining in the transfected cell line. These findings show that the presence of HBV DNA appears to cause a significant up‐regulation of the TGF‐α gene. This effect on the TGF‐α gene may be a mechanism by which HBV contributes to the etiology of hepatocellular carcinoma in some patients. © 1992 Wiley‐Liss, Inc.
Serum antibodies to structural proteins of hantavirus arise at different times after infection
Authors: Groen J. , Dalrymple J. , Fisher‐Hoch S. , Jordans J. G. M. , Clement J. P. , Osterhaus A. D. M. E.
Published: March 2007

An enzyme‐linked immunosorbent assay (ELISA) was developed for the quantification of serum antibodies against group‐specific epitopes of the glycoproteins (G1, G2) and nucleoprotein (NP) of the genus Hantavirus. This assay was used to study the kinetics of the development of serum antibodies after natural infection with Puumala‐like virus in humans. To this end a panel of 34 serum samples collected from individuals at different times after natural infection was tested by the ELISA. The samples were also tested for specific IgM and IgG levels against Puumala‐like virus, which provided confirmatory data about the presumed timing of infection. It was shown that serum antibodies against the G1 epitope were present in the acute and early convalescent period just before antibodies to the NP epitope could be demonstrated. In contrast, antibodies to two G2 epitopes were present not earlier than in the convalescent and late convalescent period. Since all these categories of antibodies seem to persist for long periods, antibodies against the G1 epitope and the NP epitope may be of specific diagnostic value. Furthermore, levels of G1‐specific antibodies and antibodies to either NP or G2 may allow estimation of the time elapsed following initial infection. © 1992 Wiley‐Liss, Inc.
Prevalence of hepatitis‐C virus infection in children with chronic post‐transfusion hepatitis
Authors: Puchhammer‐Stöckl Elisabeth , Hofmann Hanns , Fink Franz Martin , Mor Wolfgang , Höcker‐Schulz Sonja , Heinz Franz‐Xaver , Kunz Christian
Published: March 2007

The prevalence of hepatitis‐C virus (HCV) infection was investigated in a group of children with chronic post‐transfusion hepatitis using a first‐and second‐generation HCV‐antibody ELISA, 2 confirmatory tests (a second‐generation recombinant immunoblot assay and a line immunoas‐say) as well as an HCV‐polymerase chain reaction (PCR). In 33% of the children, clear discrepancies were observed between the 4 different HCV‐antibody detection assays, indicating that the serological diagnosis of HCV infection is still problematic.

HCV RNA was detectable by PCR in only 69% of the antibody positive patients, which may be due to a fluctuation of viraemia during the course of infection. Such a fluctuation was demonstrated in 6 patients from whom serum samples drawn at different times were investigated. In contrast, in 8 of the 15 seronegative patients, HCV infection was identified only by PCR, although the hepatitis had already persisted for more than 2 years.

Antibody assays and PCR together detected HCV infection in about 90% of the patients with chronic hepatitis. When markers of hepatitis B infection were also investigated, only 6% of the cases remained undiagnosed. © 1992 Wiley‐Liss, Inc.
Evidence for T‐cell involvement during the acute phase of echo virus meningitis
Authors: Lucht Frédéric , Cordier GenevièVe , Pozzetto Bruno , Frésard Anne , Revillard Jean‐Pierre
Published: March 2007

Little is known about the cellular immune response during the acute phase of enterovirus infection. This was studied in patients with echovirus meningitis by analysing changes in the lymphocyte subset distribution both in blood and in cerebrospinal fluid (CSF) and their stage of activation. A significant increase of whole T‐cell and CD4 + T‐cell counts was observed in CSF in parallel with a slight decrease of T‐cell percentage in blood. Activation of the cellular immune response is supported by the observation of elevated neopterin levels in serum and CSF. However, the phenotypic markers of T‐cell activation, IL‐2 receptor (CD251, and HLA‐DR antigens were not detected in blood or in CSF. © 1992 Wiley‐Liss, Inc.
Case report: Detection of varicella‐zoster virus DNA in maternal breast milk
Authors: Yoshida Masami , Yamagami Nazuko , Tezuka Tadashi , Hondo Ryo
Published: March 2007

A 27‐year‐old woman developed chickenpox postpartum. Her 2‐month‐old son who was breast fed also acquired chickenpox 16 days after the onset of the mother's illness. Varicella‐zoster virus (VZV) DNA was detected in maternal breast milk by the polymerase chain reaction. These results suggest that transmission of VZV may occur via maternal breast milk. © 1992 Wiley‐Liss, Inc.
Polymerase chain reaction assay for hepatitis C virus RNA using a single tube for reverse transcription and serial rounds of amplification with nested primer pairs
Authors: Lin Hsiang Ju , Shi Naiyi , Mizokami Masashi , Hollinger F. Blaine
Published: March 2007

A procedure is described for the detection of hepatitis C virus (HCV) RNA in blood by means of the polymerase chain reaction (PCR) in which the reverse transcription step and two rounds of amplification are carried out in a single tube. This results in fewer manipulations, reduced risk of contamination, and economy of time. The procedures are generally applicable to other assays based on the PCR. We describe the preparation (from 100 μL serum) of test samples that remain stable for at least 6 days under specified conditions and an assay that employs nested primer pairs homologous to conserved sequences in the 5′ noncoding region. The method was tested on 107 sera from the United States and Japan. Correlation with first‐generation anti‐HCV was 77%. Two sets of nested primer pairs homologous to sequences in the 5′ noncoding region and one set based on structural region sequences showed differences in their reactivities with serum HCV RNA. The recommended single tube procedure specified a primer for reverse transcription that was conserved in all reported HCV genomes but absent from pestivirus genomic sequences. The effects of preanalytical factors on the detection of HCV RNA were studied. Qualitatively, there was no change in the HCV RNA‐positivity of sera that were exposed to room temperature for 24 hours. Quantitative studies showed a decrease in titer in some specimens. Three cycles of freeze‐thawing had no detectable effects on the titers of HCV RNA. © 1992 Wiley‐Liss, Inc.