Evaluation of telomerase activity in non-genital Bowen's disease.

We investigated the level of telomerase activity (TA) in 17 specimens of non-genital Bowen's disease (BD) and in 14 specimens of skin without sun exposure (non-exposed skin) using a non-isotopic PCR-based telomeric repeat amplification protocol (TRAP) assay. Expression of human telomerase reverse transcriptase (hTERT; the catalytic subunit of telomerase) was also evaluated by immunochemistry in the non-genital BD tissues. Moderate to high levels of TA were detected in 41.2% of 17 non-genital BD specimens (P = 0.001). In contrast, TA was not evident in non-exposed skin. Recently, nucleolin was reported to be associated with hTERT, so we used this antibody instead of hTERT antibody. Immunohistochemistry showed that nucleolin expression was associated with high TA levels in non-genital BD. Our results also revealed differences of TA levels among non-genital BD specimens. High levels of TA in those specimens were not age related. Five out of 7 specimens (71.4%) with moderate to high TA levels were from sun-exposed sites, while the remaining 10 specimens with low levels of TA were from non-exposed sites. These results suggested that cellular DNA damage caused by ultraviolet irradiation might be associated with an increase of TA in non-genital BD. Among non-genital BD specimens, 4 out of 17 (23.5%) showed high levels of TA (median relative TA value: 79.8%; P = 0.003), which might be associated with immortalization or transformation to invasive squamous cell carcinoma.

The effects of topical application of phytonadione, retinol and vitamins C and E on infraorbital dark circles and wrinkles of the lower eyelids.

BACKGROUND: Infraorbital dark circles and wrinkles of the lower eyelids are a cosmetic problem, especially with age. AIMS: To determine whether a gel containing 2% phytonadione, 0.1% retinol and 0.1% vitamins C and E is effective in reducing dark under-eye circles and wrinkles of the lower eyelids in healthy Japanese adults. PATIENTS/METHODS: Fifty-seven adult Japanese volunteers with dark under-eye circles and wrinkles were enrolled in an open label study. The gel formulation was applied twice daily to the lower eyelid site for 8 weeks. Haemostasis, pigmentation and wrinkles were evaluated by a physician and by the patients themselves, using a digital camera and a visual analogue scale respectively, after 4 and 8 weeks of treatment. RESULTS: Topical application of the gel decreased not only haemostasis but also wrinkles after 8 weeks of treatment. Of 57 patients, 27 (47%) had reductions in haemostasis. Wrinkles were also decreased in some patients. However, pigmentation was not clearly removed by this gel. CONCLUSIONS: Topical application of the gel containing 2% phytonadione, 0.1% retinol, 0.1% vitamin C and 0.1% vitamin E was fairly or moderately effective in reducing dark under-eye circles, especially in cases of haemostasis, over a short treatment period in healthy Japanese adults. This treatment also slightly decreased wrinkles.

Human papillomavirus associated Bowen's disease of the foot: unique clinical features mimicking a common wart.

We describe a case of Bowen's disease that developed on the left dorsal foot of a 56-year-old female patient and that mimicked the clinical features of a common wart. PCR-based analysis indicated that the lesion contained HPV-16 DNA and HPV-16 positive cells were found in the upper layers of the epidermal lesion using in situ hybridization with an HPV-16 probe.

Human papillomavirus type 58 in Bowen's disease of the elbow.

Human papillomavirus (HPV) can be detected in skin lesions of Bowen's disease, particularly on the fingers, and its genotype is associated with mucosal/genital types of HPV. We report herein an 85-year-old woman who had HPV-associated Bowen's disease on her elbow. HPV-58 DNA was detected in the lesion by polymerase chain reaction with restriction fragment length polymorphism and by Southern blot hybridization. In situ hybridization revealed numerous hybrid cells in the nuclei of the upper epidermis and stratum corneum of Bowen's disease. A high-risk type of mucosal HPV-58 DNA is associated with Bowen's disease in this case, suggesting that HPV-related Bowen's disease is not always restricted to genital or finger lesions.

The presence of mucosal human papillomavirus in Bowen's disease of the hands.

BACKGROUND: Bowen's disease (BD) of the genital skin region is generally associated with human papillomavirus (HPV) infection. Various molecular analyses have identified mainly HPV-16 in the lesions. However, the HPV genotypes associated with BD of the hands have not yet been characterized. METHODS: The skin specimens of 12 patients with BD of the hands were investigated clinicopathologically and immunohistochemically, and the total DNAs extracted from the skin were analyzed for the presence of HPV DNA using Southern blot hybridization and polymerase chain reaction (PCR) in combination with restriction fragment length polymorphisms (RFLP) and direct sequencing analysis of the amplified products. In addition, the histologic localization of HPV DNA was examined by in situ hybridization in paraffin embedded sections of HPV positive patients. RESULTS: In 8 of 12 BD lesions (66.7%), HPV types (HPV-16, -31, -54, -58, -61, -62, and -73) were detected by Southern blot hybridization and/or PCR with RFLP and direct sequencing analysis. In 6 of 7 HPV positive lesions examined (85.7%), the viral genomes were identified by in situ hybridization in the nuclei of keratinocytes in the upper stratum malpighii and/or stratum corneum. CONCLUSIONS: These results demonstrate that BD of the hands is frequently associated with HPV infection. The seven HPV genotypes are known as mucosal (genital) HPVs and to the authors' knowledge, this is the first time HPV-31, -54, -58, -61, -62, and -73 have been identified in BD lesions. These findings strongly suggest that HPVs related to mucosal lesions play an important role in the development of BD of the hands.

[Effects of shi-ka-ron and Chinese herbs in mice treated with anti-tumor agent mitomycin C].

The Shi-ka-Ron, and its constituent Chinese herbs Lithospermum erythrorhizon, Astragalus membranaceus and Ligusticum Wallichii were administered with antitumor agent, mitomycin C (MMC) to ICR mice, and their effects on murine macrophages and lymphocytes were studied. Peritoneal macrophages were significantly inhibited both in their number and chemotactic activity by MMC treatment. Splenic weight and blastogenic responsiveness to Concanavalin A of spleen lymphocytes also decreased significantly in MMC-treated mice. NK cell activity was also suppressed by MMC treatment. When these mice were orally treated with extracts of Shi-ka-ron or each Chinese herbs mentioned above, it showed protective effects to immunosuppressive mice on all 5 items studied. The number of macrophages, and the functions of macrophages and lymphocytes maintained the same or more than normal levels in MMC plus each group of these extracts treated mice. These results suggest that the Shi-Ka-Ron and Chinese herbs could resist immunosuppression induced by antitumor agent MMC, and its mechanisms might be correlated with stimulation of the RES (reticuloendothelial system), activation of T cell blastogenesis and NK cell cytotoxicity.

[Immunomodulative effects of Chinese herbs in mice treated with anti-tumor agent cyclophosphamide].

Extracts of Chinese herbs were administered with antitumor agent, cyclophosphamide (CY), and their effects on macrophages and lymphocytes were studied. Number of peritoneal macrophages significantly decreased and their chemotactic activity was suppressed by treatment with CY. Blastogenic responsiveness to Concanavalin A and NK cell activity of spleen lymphocytes were suppressed significantly in CY-treated mice. Extracts of Lithospermi radix, Astragali radix and Glycyrrhizae radix showed protective effects on immunosuppressive mice. The number of macrophages, chemotactic activity of macrophages and blastogenic response of lymphocytes were recovered to the same or more than that of normal levels. An extract of Ginseng radix showed protective effects on the number and functions of macrophages by treatment with CY but did not show any effects on the lymphocytic blastogenesis. On the contrary it showed a strong inhibitory effect on the NK cell activity. These results suggest that Chinese herbs could modulate cellular immune response, especially in the activation of macrophages and splenic lymphocytes.

Effect of shi-ka-ron and Chinese herbs on cytokine production of macrophage in immunocompromised mice.

Shi-Ka-Ron is a prescription composed of 8 crude extracts of Chinese herbs. It reduces suppression of cytokine production by peritoneal macrophages in mice Immunocompromised by the anti-tumor agent, cyclophosphamide (CY), in vivo. Although it dose not increase IL-1 production in vitro, it enhances TNF production. We found that Ginseng radix, Lithospermi radix, Astragli radix and Glycyrrhizae radix somewhat reduced suppression of cytokine production in CY treated macrophages. Especially, Glycyrrhizae radix shows an active immune response both in vivo and in vitro. Our results suggested that the mechanism underlying immunomodulation of Shi-Ka-Ron is closely related to cytokine production: each herb stimulating macrophages.

Immunological properties of TtT/M-87 cell line established from murine pituitary tumor-associated macrophages.

TtT/M-87 cell is a macrophage cell line established from thyrotropic pituitary tumor tissues in mouse. In this paper, we report the immunological properties of M-87 cells as a model of tumor-associated macrophage. Contrasting with resident peritoneal macrophages, M-87 cells constitutively secreted small but significant amounts of TNF-alpha and IL-1 alpha, which were detectable in both biological assays (cytotoxic activity for L929 and co-mitogenic activity for Con A-induced T cell proliferation, respectively) and ELISA, and produced larger amounts of these cytokines upon stimulation with LPS. They expressed MHC class II molecules on their cell surface without stimulation by IFN-gamma. The accessory or antigen-presenting cell activity in antibody-producing response of spleen lymphocytes to sheep red blood cells was shown to be much higher in M-87 cells than normal peritoneal macrophages. In addition, when normal spleen lymphocytes were cultured with allogeneic tumor cells, such as EL-4 and S-180, in the presence of M-87 cells, lymphocytes reactive to stimulator cells were activated to manifest inhibitory effect on the tumor cell growth and also to manifest specific cytotoxic effect on the allogeneic tumor cells. These results show that M-87 cells derived from tumor-associated tissue are activated macrophages and that they are inhibitory to tumor cell growth and augmentative in the induction of T-cell-mediated immune responses.