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Biomedical subjects

K Fujinaga

Publications and source records attributed to K Fujinaga.

At least 127 records · Page 7Linked to original sources
PubMed1991

Relatively low prevalence of human papillomavirus 16, 18 and 33 DNA in the normal cervices of Japanese women shown by polymerase chain reaction.

Ninety-two cervical scrapes and tissues, obtained from cytologically or histologically normal cervices of Japanese women, were examined for the presence of human papillomavirus (HPV) 16, 18 and 33 DNA by the polymerase chain reaction (PCR) method. Five out of 92 cases were HPV 16 DNA-positive, but neither HPV 18 nor 33 DNA was detected. The HPV (type 16, 18 and 33) prevalence rate in pregnant women, including postpartum, was 10% (3/31), which was higher than that in non-pregnant women. In two HPV 16-positive cases, we detected HPV 16 DNA again 2 months later. HPV (type 16, 18 and 33) prevalence in normal cervices was shown to be relatively low. However, it is very important to follow up the HPV-positive cases in cytologically normal cervices in order to elucidate the relation between HPV infection and the progression of cervical cancer.

Adolescent↗
PubMed1991

Fastidious human adenovirus type 40 can propagate efficiently and produce plaques on a human cell line, A549, derived from lung carcinoma.

Human adenovirus type 40 (Ad40) cannot propagate in conventional established human cell lines such as KB or HeLa cells. However, it has been shown that Ad40 DNA replicates in KB18 cells which express Ad2 E1B genes, suggesting that Ad40 is defective in the E1B gene function in KB or HeLa cells. We show here that Ad40 can propagate and produce plaques on A549 cells which do not contain Ad E1B genes. Our experiments show that the levels of replication of Ad40 DNA and production of infectious Ad40 virus in A549 cells are the same as or higher than those in 293 or KB18 cells. Dot blot analysis shows that the levels of Ad40 E1A and E1B mRNAs expressed in A549 cells at early to intermediate times postinfection are at least 10-fold higher than those in KB or KB18 cells. Northern (RNA) blot analysis shows that large E1B mRNA species (approximately 24S to 26S) are synthesized prior to the onset of DNA replication in A549 cells. No E1B mRNA species are synthesized in KB or KB18 cells at early times postinfection, and no differences in the expression of E1B mRNAs are seen between KB and KB18 cells. The experiment suggests that A549 cells have a cellular factor(s) which activates Ad40 E1B mRNA synthesis and that the E1B mRNA synthesis helps Ad40 propagation. In contrast, Ad40 can propagate in KB18 cells by using Ad2 E1B gene products that are constitutively expressed in this cell line. Furthermore, this result shows that Ad40 cannot propagate in KB cells because of the failure in the expression of E1B genes at early times postinfection.

Adenovirus Early Proteins↗
PubMed1991

Outbreak of pertussis in a residential facility for handicapped people.

An outbreak of pertussis was recognized and investigated in a ward for neurologically impaired residents. Of 50 residents ranging in age from 8 to 25 years old, 41 (82%) had serological or bacteriological evidence of pertussis infection and 28 (56%) developed typical symptoms of pertussis. Of the 50 residents, nine were immunized and 41 were unimmunized. There were no significant differences in the rate of infection and rate of developing typical symptoms between the immunized and unimmunized (9/8/4 vs 41/33/24). Of 43 carers, 6 (14%) developed typical symptoms and four of the six showed serological evidence of pertussis infection. Epidemiological survey of this outbreak revealed that pertussis was introduced and spread by the carers. Erythromycin which was administered only to the residents did not prevent the spread of pertussis. We conclude that pertussis immunization including a booster dose should be considered for neurologically impaired people in a closed environment and erythromycin prophylaxis should include adult carers.

Adolescent↗
PubMed1991

[A case report of Buschke-Lowenstein tumor (giant condyloma)].

A 28-year-old woman with diabetes mellitus and alcoholic hepatitis presented a rare case of Buschke Lowenstein tumor, or giant condyloma of vulva. HPV DNA of this tumor was detected by in situ hybridization using tritium labeled HPV 6 b, 16 and 18 DNA. This tumor appeared to harbour HPV 6 b DNA, but other HPV DNAs were negative. The distribution of HPV 6 b DNA was detected in the nucleus of the squamous epithelium showing koilocytosis. Total vulvectomy and resection of the clitoris were done. The postoperative course was uneventful and there has been no recurrence of tumor so far.

Adult↗
PubMed1990

The physical state of human papillomavirus 16 DNA in cervical carcinoma and cervical intraepithelial neoplasia.

Cervical carcinomas and cervical intraepithelial neoplasias (CIN) were analyzed for the presence of human papillomavirus (HPV) DNA using Southern blot hybridization. Of the five HPV types examined (HPV types 6, 11, 16, 18, and 33), HPV 16 DNA was detected most frequently. In most HPV 16-positive carcinomas examined, HPV 16 DNA was present in an integrated state in cellular DNA with or without the coexistence of episomal species. In one case, however, only episomal species were detected. Among seven cases of HPV 16-positive CIN, four contained HPV 16 DNA only in the episomal state and the rest contained HPV 16 DNA only in the integrated state, but the coexistence of both states was not found. These results suggest that the integration of HPV 16 DNA is not necessary for cells to become malignant, although it is frequently associated with malignant cells.

Blotting, Southern↗
PubMed1990

Detection of adenovirus type41 in stool samples by a latex agglutination method.

We have developed a simple agglutination (LA) method for the detection of enteric adenovirus (EAd) in stool samples from infants with acute gastroenteritis. Ad type 41 (Ad41) was detected with high sensitivity and specificity by a slide agglutination test using latex particles coated with antiAd41 antibody (LA-antiAd41). The agglutination of LA-antiAd41 with Ad41 on a glass slide was evident macroscopically within 2 min. The sensitivity of the LA method was four times higher than that of the EM method.

Adenoviruses, Human↗
PubMed1990

Identification of sequences responsible for positive and negative regulation by E1A in the promoter of H-2Kbm1 class I MHC gene.

The mechanism of transcriptional regulation of the H-2Kbm1 major histocompatibility complex (MHC) class I gene by adenovirus type 12 E1A (Ad12-E1A) was studied in transfected rat embryonal fibroblasts. Results of long-term expression of the chloramphenicol acetyl transferase (CAT) gene placed under the control of the 5'-flanking region of the mouse MHC class I gene. H-2Kbm1, and the results of nuclear run-on transcription assays, yield evidence for both positive and negative regulation of H-2Kbm1 by E1A gene product. Deletion studies in the H-2Kbm1 promoter region revealed that a proximal 58 bp upstream sequence (-194 to -136, relative to the cap site) and a distal 316 bp sequence (-1837 to -1521) respectively contribute to positive and negative regulation mediated by the E1A gene product. Both regulatory elements of MHC class I gene promoter region are responsible for the differential expression of the H-2Kbm1 gene in Ad12 transformed cells. A nuclear factor binding to the negative element has been detected only in extracts derived from cells expressing Ad12-E1A.

Adenovirus Early Proteins↗
PubMed1990

Suppression of interferon-induced oligo-2',5'-adenylate synthetase induction in persistent infection.

Persistent infections with several strains of mumps virus (strains Torii and Miyahara), measles virus (strains Edmonston, CAM-70, AIK-C and Schwarz) and subacute sclerosing panencephalitis (SSPE) virus (Hälle and Mantooth) were established in various cell lines (FL, KB, A549, SK-AS, 293, K562, Ramos and NC-37). Oligo-2',5'-adenylate synthetase activity was demonstrated to be only slightly induced by interferon in cytoplasmic and nuclear fractions of cell lines persistently infected with mumps virus. In these cells, resistance to vesicular stomatitis virus infection was not induced by interferon treatment. Treatment of the persistently infected cells with interferon for 10 and 24 h did not stimulate an increase in the amount of synthetase mRNA. In cells persistently infected with measles and SSPE viruses, reduced induction of the enzyme varied with host cell types. Induction of the enzyme was not found in K562, SK-AS and KB cells, but was recognized in NC-37 and FL cells.

2',5'-Oligoadenylate Synthetase↗
PubMed1990

Amplification and specific detection of transforming gene region of human papillomavirus 16, 18 and 33 in cervical carcinoma by means of the polymerase chain reaction.

We have established a highly sensitive method for specific detection of human papillomavirus (HPV) 16, 18 and 33, by using the polymerase chain reaction (PCR). A HPV-related sequence (140 bp) in the E6 transforming region was specifically amplified and detected by gel electrophoresis and by the use of a specific oligonucleotide probe. The PCR could detect 10(5)-10(6) copies per cell (maximum sensitivity). Furthermore, HPV 16, 18 and 33 DNAs were synthesized in a common reaction solution and specifically detected by HPV type-specific probes. The PCR detected the HPV sequence from tissues which were negative to Southern hybridization. This detection technique may contribute significantly to the precise analysis of HPV in small proliferative lesions in the cervix.

Base Sequence↗
PubMed1990

[Studies of human papillomavirus (HPV) in urological tumors].

Urological tumors were examined for the presence of human papillomavirus (HPV) DNA by using Southern blot hybridization. In 20 male patients with condyloma acuminatum, HPV type 6 was found at 85% (17/20), HPV type 11 at 95% (19/20), HPV type 16 at 5% (1/20) and HPV type 18 at 0% (0/20). In 2 female patients with condyloma acuminatum, HPV types 6, 11, 16 and 18 were found at 100% (2/2), 100% (2/2), 50% (1/2) and 0% (0/2), respectively. All 6 of the patients who were positive for HPV type 6, were also positive for HPV type 11. Two patients were positive for HPV types 6, 11 and 16, the last of which was frequently found in penile cancer and uterine cervical cancer. In 6 patients with penile cancer, two patients were positive for HPV type 16 and negative for HPV types 6, 11 and 18. The remaining 4 patients were negative for all these HPV types. One patient who was positive for HPV type 16 had penile cancer after three previous episodes of penile condyloma acuminatum. From this information, a malignant change in the condyloma acuminatum was assumed to indicate the possible association of HPV type 16 with the process of malignant degeneration. HPV types, 6, 11, 16 and 18 were not detected in a female patient with vulvar cancer. Although HPV was thought to participate in the development of urological tumors except for external genital tumors, all patients examined, consisting of 2 with benign prostatic hypertrophy, 5 with prostatic cancer and 24 with bladder cancer, were negative for HPV types 6, 11, 16 and 18. Eight patients with bladder cancer were negative for HPV type 33.

Adult↗
PubMed1990

[The sensitivity of in situ hybridization for detection of human papillomavirus].

We studied a sensitivity of HPV DNA detection by in situ hybridization method using 3H labeled HPV DNA. The materials were CaSki cells and SiHa cells which were derived from as a negative control. The total cellular DNAs extracted from these cell lines were estimated copy numbers of HPV 16 DNA using Southern blot hybridization. In our result, CaSki cell has 400 copies/cell, SiHa cell were appeared to have 1-5 copies/cell. Simultaneously these cells were fixed by periodate-buffered lysine-paraformaldehyde-glutaraldehyde (PLPG) and were detected HPV 16 DNA using in situ hybridization. We detected HPV 16 DNA in CaSki cells and SiHa cells by in situ hybridization also. We concluded that the sensitivity of our in situ hybridization technique is 1-5 copies/cell.

DNA, Viral↗
PubMed1989

Binding sites of HeLa cell nuclear proteins on the upstream region of adenovirus type 5 E1A gene.

Twenty one binding sites of HeLa cell nuclear proteins were identified on the upstream region of adenovirus type 5 E1A gene using DNase I footprint assay. The proximal promoter region contained five binding sites that overlapped the cap site, TATA box, TATA-like sequence, CCAAT box, and -100 region relative to the E1A cap site(+1). The -190 region was a potential site for octamer-motif binding proteins, such as NFIII and OBP100. An upstream copy of the E1A enhancer element 1 was the site for a factor (E1A-F) with the binding specificity of XGGAYGT (X = A, C; Y = A, T). E1A-F factor also bound to three other sites, one of which coincided with the distal E1A enhancer element. The distal element also contained a potential site for ATF factor. The adenovirus minimal origin of DNA replication competed for DNA-protein complex formation on the CCAAT and TATA box region and the -190 region, suggesting that these regions interacted with a common or related factor.

Adenovirus Early Proteins↗
PubMed1989

A pertussis toxin-sensitive GTP-binding protein plays a role in the G0-G1 transition of rat hepatocytes following establishment in primary culture.

Acute spontaneous c-myc gene expression and sustained increase of a GTP-binding protein(s) (G-protein) which is sensitive to islet-activating protein (IAP), pertussis toxin, occurred early during primary culture of adult rat hepatocytes. Following these earlier events, DNA synthesis was demonstrated in response to EGF and insulin. Addition of IAP immediately after plating of primary cultures inhibited c-myc expression and the hormone-induced DNA synthesis. Addition at 24 h or later following cell inoculation, however, produced only weak effects on DNA synthesis, even though the IAP-sensitive G-proteins were completely inactivated. We conclude that the IAP-sensitive G-protein(s) plays a role in the earlier process(es) of the G0-G1 transition, which is essential for the initiation of growth factor-dependent DNA synthesis.

Adenosine Diphosphate Ribose↗
PubMed1989

H-ras gene is expressed at the G1 phase in primary cultures of hepatocytes.

The expression of c-H-ras and proliferating cell nuclear antigen (PCNA) in primary cultures of rat hepatocytes was determined in order to elucidate the relationship between the c-H-ras gene and the S phase of the cell cycle. In cells treated with EGF, elevation of c-H-ras expression was detected at the 22nd, 34th, 44th, and 54th h after plating, PCNA expression and DNA synthesis were detected at the 44th and 54th h. In cells without EGF treatment, only c-H-ras expression was detected at the 44th and 54th h. In our previous report, we showed that c-myc expression increased within several hours after plating, suggesting that isolated hepatocytes traverse from G0 to G1 under culture conditions, regardless of EGF treatment. These results clearly showed that the c-H-ras gene of adult rat hepatocytes was expressed in the mid-to-late G1 phase of the cell cycle as well as in the early S phase in primary culture.

Animals↗