bluuebunny/crossref_metadata_2025_split huggingface dataset:
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

AbstractA 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

AbstractHIV‐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

AbstractGenome 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

AbstractSome 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

AbstractThe 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

AbstractSera, 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

AbstractA 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

AbstractEpstein‐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

AbstractTwo 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

AbstractThe 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

AbstractCytomegalovirus (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.
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

AbstractHaemophilic 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

AbstractA 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

AbstractRNA 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

AbstractClinically 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.