Tsuyoshi Mitsuishi

Human papillomavirus infection in actinic keratosis and bowen’s disease: comparative study with expression of cell-cycle regulatory proteins p21waf1/cip1, p53, pcna, ki-67, and bcl-2 in positive and negative lesions

We examined the presence of human papillomavirus (HPV) DNA in tissues of premalignant skin lesions, i.e., actinic keratosis (n = 13) and Bowens disease (n = 62), taken from 69 Japanese immunocompetent and renal transplant recipient patients. Detection and typing of HPV DNA were performed using polymerase chain reaction (PCR) and sequence analysis or restriction fragment length polymorphism (RFLP) analysis, respectively. The positivity rates of HPV DNA in tissues of actinic keratosis and Bowens disease were 77% and 65%, respectively. Twenty-seven HPV types were detected in 50 (67%) premalignant skin lesions, in which Z95963 (accession no. in the EMBL Databank), Z95968, AJ010823, and AJ000151 have been described as partial sequences of unknown HPV types. Furthermore, 2 unknown types, HPVX1 and HPVX2, were found in specimens of actinic keratosis. Sequence analysis showed that HPVX1 is related to HPV-37 (86.1% sequence homology) and that HPVX2 is related to HPV-38 (79.7%). These results indicate that various mucosal and epidermodysplasia verruciformis-related HPV types are associated with the pathogenesis of actinic keratosis and Bowens disease. In addition, 24 specimens of HPV-positive or HPV-negative premalignant skin lesions were examined immunohistochemically for proliferating cells to determine biological differences between HPV-positive and HPV-negative lesions. Immunohistochemistry for p21(Waf1/Cip1), p53, proliferating cell nuclear antigen (PCNA), Ki-67, and Bcl-2 revealed that there was no significant difference in the cell proliferation activity between HPV-positive and HPV-negative lesions, suggesting that HPV infection alone does not induce cell proliferation in those lesions.

Activation of STAT5 confers imatinib resistance on leukemic cells through the transcription of TERT and MDR1.

We used two imatinib resistant cell lines, K562-ADM cells, which over-express P-glycoprotein (a product of the ABCB1 gene, more commonly known as MDR1), and K562-hTERT cells, which over-express the telomerase reverse transcriptase (TERT), as models to show that the acquisition of multidrug resistance in CML is associated with the enhanced phosphorylation of signal transducer and activator of transcription 5 (STAT5). The induction of P-glycoprotein expression that occurred in response to adriamycin treatment was accompanied by increased phosphorylation of BCR-ABL and STAT5, as well as increased telomerase protein expression. Intriguingly, a ChIP assay using an anti-STAT5 antibody revealed direct binding of STAT5 to the promoter regions of both the human TERT gene and the MDR1 gene in K562-ADM cells. Conversely, silencing of endogenous STAT5 expression by siRNA significantly reduced both the expression of P-glycoprotein and telomerase activity and resulted in the recovery of the imatinib sensitivity of K562-ADM cells. These findings indicate a critical role for STAT5 in the induction of P-glycoprotein and in the modulation of telomerase activity in drug-resistant CML cells. Furthermore, primary leukemic cells obtained from patients in blast crisis showed increased levels of phospho-STAT5, P-glycoprotein and telomerase. In contrast, none of these proteins were detectable in the cells obtained from patients in the chronic phase. Together, these findings indicate a novel mechanism that contributes toward multidrug resistance involving STAT5 as a sensor for cytotoxic drugs in CML patients.

Treatment of localized epidermodysplasia verruciformis with tacalcitol ointment

A 69‐year‐old Japanese woman presented with multiple, erythematous, keratotic plaques distributed on her right breast, right shoulder, and left cubital fossa. The lesions varied in size from 5 mm to 6 cm. The patient had first noticed a small area of primary erythema on her left cubital fossa at the age of 18 years. The number and size of the lesions had increased for 50 years. A black nodule appeared on the plaque on the right breast 3 years ago and enlarged to 5 cm in diameter ( Fig. 1A ).

Novel Polymerase Chain Reaction Method for Detecting Cutaneous Human Papillomavirus DNA

There is no simple test to identify the human papillomavirus (HPV) genotypes that cause cutaneous warts. A new polymerase chain reaction (PCR) method, called SK‐PCR, was developed for this purpose. This PCR amplifies 210–238 base pairs of L1 DNA of 17 HPV types (HPV‐1a, ‐2a, ‐3, ‐4, ‐7, ‐10, ‐27, ‐28, ‐29, ‐40, ‐57, ‐60, ‐63, ‐65, ‐77, ‐91, and ‐94), which are thought to cause various cutaneous warts, including common, flat, butchers, punctate, and pigmented warts. The method is novel because the location of these primers is completely different from that of any previous PCR method for HPV. The target sequences are specific to alpha‐, gamma‐, and mu‐papillomaviruses (PVs), but not to beta‐PVs. Furthermore, direct sequencing and restriction fragment length polymorphism (RFLP) were used to determine the HPV genotypes. Fifty of samples of plantar warts were examined, and HPV‐27 was identified in 22 warts, HPV‐57 in 15 warts, and HPV‐2a in 9 warts. These PVs, which are alpha species 4, were the most common. HPV‐4 and ‐65 (gamma‐PVs) and HPV‐1a and ‐63 (mu‐PVs) were detected in one case each. A single HPV type was identified in all of these warts. This method appears to be useful for genotyping the HPVs causing skin warts, and for distinguishing between HPV‐induced warts and warty lesions unrelated to HPV infection. J. Med. Virol. 84:138–144, 2011.

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.

Akt and PKC are involved not only in upregulation of telomerase activity but also in cell differentiation-related function via mTORC2 in leukemia cells

We have shown previously that PI3K/Akt pathway is active after cell differentiation in HL60 cells. In the present study, we have investigated whether additional molecules, such as protein kinase C (PKC), are involved in the regulation, not only of telomerase, but also of leukemia cell differentiation. We show that PKC activates telomerase and is, itself, activated following VD3- or ATRA-induced differentiation of HL60 cells, as was observed for PI3K/Akt. To clarify the significance of PI3K/Akt and PKC pathway activation in leukemia cell differentiation, we examined the active proteins in either the downstream or upstream regulation of these pathways. In conjunction with the activation of Akt or PKC, mTOR and S6K were phosphorylated and the protein expression levels of Rictor were increased, compared with Raptor, following cell differentiation. Silencing by Rictor siRNA resulted in the attenuation of Akt phosphorylation on Ser473 and PKCα/βII phosphorylation, as well as the inhibition of Rictor itself, suggesting that Rictor is an upstream regulator of both Akt and PKC. In addition, in cells induced to differentiate by ATRA or VD3, Nitroblue-tetrazolium (NBT) reduction and esterase activity, were blocked either by LY294002, a PI3K inhibitor, or by BIM, a PKC inhibitor, without affecting cell surface markers such as CD11b or CD14. Intriguingly, the silencing of Rictor by its siRNA also suppressed the reducing ability of NBT following VD3-induced cell differentiation. Taken together, our results show that Rictor associated with mTOR (mTORC2) regulates the activity of both Akt and PKC that are involved in cell functions such as NBT reduction and esterase activity induced by leukemia cell differentiation.

Epidermodysplasia verruciformis with keratoacanthoma, Bowen's disease and squamous cell carcinoma: isolation of high‐risk types of HPV 5 and unknown type of human papillomavirus

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Poroid hidradenoma: A case report with review of Japanese published work

Dear Editor, We report a case of poroid hidradenoma (PH) arising on the leg. A 34-year-old Japanese male presented with a slowly enlarging nodule on his left leg which had been present for 6 years. Physical examination revealed a dark bluish, dome-shaped nodule, 21 mm × 13 mm in size, on the left side of the anterior aspect of his left leg (Fig. 1). Excisional biopsy was performed under local anesthesia. The excised specimen was embedded in paraffin, and serial sections were stained with hematoxylin–eosin, and immunohistochemical studies with antibodies for cytokeratin (CK) and carcinoembryonic antigen (CEA) were also performed. Histopathological examination showed that the neoplasm was a well-demarcated cyst with small solid components protruding into the cystic space (Fig. 2). There was no apparent connection to the overlying epidermis. At higher magnification, the tumor nests appeared to be composed of two types of cells: small and dark-staining poroid cells, and large and paler cuticular cells. Vacuolar spaces were seen within the cytoplasm of some cuticular cells. Ductlike structures surrounded by the cuticular cells were also observed (Fig. 3). Sclerotic stroma contained blood vessels. There was no evidence of decapitation secretion. These findings were consistent with poroid hidradenoma. Immunohistochemical findings were as follow. The lining cells of the duct-like structures and some of the surrounding cuticular cells were stained with antibody 35βH11 (DAKO Japan, Kyoto, Japan) against CK 8, which is expressed in the eccrine secretory elements and occasionally in the inner surfaces of dermal eccrine ducts. Some poroid cells and cuticular cells of the neoplasm were occasionally positive to antibody DE-K10 (DAKO Japan) against CK 10, which suggests the differentiation of the intermediate cells of the eccrine ducts. CEA was positive in the cells adjacent to the duct-like structures. CEA is presented in the inner cells of the eccrine ducts and the inner surfaces of the eccrine secretory components.

Human Papillomavirus Type 7-Associated Condyloma

Human papillomavirus type 7 (HPV-7) was originally identified in common warts of butchers. It has remained unclear, however, whether HPV-7 also induces other distinct types of cutaneous lesions. We observed similar keratoses on the groins of 2 patients. The lesions presented as multiple, smooth and small with little change in color from that of the adjacent skin and were diagnosed as condylomas. Their histological features were acanthosis, papillomatosis and parakeratosis with hyperkeratotic perinuclear vacuolation in the granular layer. By Southern blot analysis, HPV-7 DNA was identified in both condylomas. We conclude that HPV-7 infection causes condyloma. In addition, we discuss the long-held dogma regarding the association of HPV-7 with butcher’s warts and highlight the potential need for clinicians to know causal HPV types in cutaneous lesions given the increased use of prophylactic HPV vaccines.

Bowen's disease of the nail apparatus and association with various high-risk human papillomavirus types

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