The prognostic value of a reverse transcriptase-PCR assay of sentinel lymph node biopsy for patients with cutaneous melanoma: a single-center analysis in Japan.

Sentinel lymph node biopsy (SLNB) is the standard of care for staging melanoma. However, limited research has been carried out on the prognostic value of SLNB in patients with primary cutaneous melanoma in Asian countries. The objective is to evaluate the efficacy of SLNB in Japanese patients with primary cutaneous melanoma and to elucidate whether reverse transcriptase (RT)-PCR analysis of sentinel lymph nodes (SLNs) is valuable for predicting patient outcome. A total of 101 patients with primary cutaneous melanoma underwent SLNB at the Department of Dermatology, Kyushu University (Fukuoka, Japan), between May 2001 and December 2009. The removed nodes were stained with hematoxylin-eosin and with immunohistochemical stains for HMB-45, tyrosinase, MART-1, and MITF and multiple-mRNA marker (MART-1, tyrosinase, and GP-100) RT-PCR assays were conducted. The following clinicopathological variables were evaluated: age, sex, histological type, tumor site, Breslow thickness, disease-free survival (DFS), and melanoma-specific survival (MSS). Several parameters were analyzed for DFS and MSS using the Kaplan-Meier method and Cox proportional hazards model. The success rate of identifying SLNs was 98% (99 of 101 cases). Tumor-positive SLNs were significantly correlated with higher Breslow thickness, stage, tumor subtype, and tumor site. Patients with tumor-positive SLNs had a significantly shorter MSS and DFS than those with tumor-negative SLNs (P=0.0153 and 0.0004, respectively). Patients with at least two positive markers in the RT-PCR assay had a significantly shorter DFS than those with less than one marker (P=0.013). SLNB and multimarker RT-PCR analysis are useful for predicting the prognosis of patients with melanoma.

Pneumatosis cystoides intestinalis with systemic sclerosis, limited type resulting in a poor prognosis.

Pneumatosis cystoides intestinalis (PCI) is a rare disease characterized by the presence of multilocular intramural clusters of gas in the alimentary tract and has been considered to have a favorable response to conservative treatment. We describe the first case of limited type of systemic sclerosis (SSc) with PCI. A 74-year-old Japanese woman presented with a 4-month history of an unhealed cutaneous ulcer on the right third finger, along with sclerodactyly of bilateral hands. Proximal skin sclerosis was absent. The patient reported acute abdominal pain, and a diagnosis of PCI was established on plain radiography. The patient died of multiple organ failure 5 months after the development of PCI. PCI is rarely complicated with SSc, and all cases previously reported were associated with diffuse SSc. Because PCI is one of the poor prognostic factors of SSc, we should recognize the presence of this condition even in patients with limited cutaneous involvement.

Expression of p16 and hTERT protein is associated with the presence of high-risk human papillomavirus in Bowenoid papulosis.

BACKGROUND: High-risk human papillomavirus (hrHPV) type E6 and E7 oncoproteins contribute to oncogenesis in multiple ways by modulating the activities of host components in cell-cycle regulation including the expression of p16 protein (p16) and human telomerase reverse transcriptase (hTERT). The expression of p16 and hTERT protein in Bowenoid papulosis (BP) has not been studied. METHODS: Biopsy samples of BP from 26 patients were subjected to in situ hybridization for various HPV strains and immunohistochemical staining for p16 and hTERT. RESULTS: Among the 26 biopsy specimens, in situ hybridization using DNA probes for HPV 16/18 revealed positivity in 18 specimens (69.2%), one of which also showed positivity with the probes for HPV 6/11. HPV 31/33/35 was found in three specimens (11.5%). Two specimens (7.7%) were positive for unclassified HPV. Twenty-one BP specimens that were infected with hrHPV were positive for p16 and/or hTERT. Moderate or strong and diffuse immunostaining was observed for p16 in 15 hrHPV-infected specimens and for hTERT in 16 hrHPV-infected specimens. The expression of p16 or hTERT was each significantly associated with the presence of hrHPV. CONCLUSIONS: hrHPVs were involved in inducing p16 and hTERT overexpression in BP. Moreover, our results suggested that immunohistochemical p16 and hTERT expression might be a useful marker of hrHPV infection in BP.

Anti-CXCR3 staining is useful for detecting human cutaneous and mucosal mast cells.

Human synovial mast cells (MC) can be immunolabelled with antihuman CXCR3 antibody (Ab) (clone 49801). We have investigated whether cutaneous and mucosal MC are stained with anti-CXCR3 Ab in paraffin-embedded sections. Immunohistochemical staining and immunofluorescence double staining assays were performed with anti-CXCR3, anti-tryptase, and anti-chymase Ab using normal skin, psoriatic skin lesions, and normal colon. When compared with tryptase and chymase staining, 100% of the cutaneous and 98% of the mucosal MC were positive for CXCR3. Anti-CXCR3 staining is a useful marker for human cutaneous and mucosal MC in paraffin-embedded sections.

Clinical evaluation of allogeneic cultured dermal substitutes for intractable skin ulcers after tumor resection.

Clinical research on allogeneic cultured dermal substitute (CDS), which was newly developed at the R&D Center for Artificial Skin of Kitasato University, has been carried out in medical centers across Japan with the support of the Millennium Project of the Ministry of Health, Labor and Welfare of Japan. Allogeneic CDS was prepared by cultivation of fibroblasts on a two-layered spongy matrix of hyaluronic acid and atelo-collagen. This paper reports the clinical results of application of allogeneic CDS in 12 patients with full-thickness skin defects after surgical resection of skin tumors. In 9 of 10 patients, healthy granulation tissue developed immediately, allowing us to perform split-thickness skin grafts at an early stage. In two cases, allogeneic CDS was used to cover an expanded mesh skin graft that had been applied to treat a large ulcer, and rapid epithelization was observed. No patient developed local infection nor local tumor recurrence after treatment with CDS. The spongy matrix itself as well as the vascular endothelial growth factor (VEGF) released by the allogeneic CDS seemed to be beneficial for the treatment of intractable skin ulcers. Allogeneic CDS functions as an excellent biological dressing, and could dramatically change the treatment of intractable skin ulcers.

Photocured, styrenated gelatin-based microspheres for de novo adipogenesis through corelease of basic fibroblast growth factor, insulin, and insulin-like growth factor I.

De novo adipose tissue formation appears to proceed via two different biological events: neovascularization and spontaneous accumulation of preadipocytes and subsequent differentiation to mature adipocytes. In this article, we perform accelerated de novo adipose tissue engineering using photocured, styrenated, gelatin-based microspheres (SGMs) with different drug release rates of immobilized angiogenic and adipogenic factors. The concept of this system is to induce neovascularization and migration of endogenous preadipocytes by the rapid delivery of the angiogenic factor basic fibroblast growth factor (bFGF), followed by the proliferation and differentiation of preadipocytes into adipocytes by the prolonged delivery of the adipogenic factors, insulin and insulin-like growth factor I (IGF-I). Bioactive substance-immobilized SGMs with different drug release rates were prepared with different gelatin concentrations. An in vitro study showed the prolonged release of an immobilized model protein and the dependence of drug release rate on gelatin concentration. After the subcutaneous injections of SGMs immobilized with these bioactive substances in different combinations, the formation of masses or clusters of adipocytes was observed in rats. Triglyceride content in the injection site for the group that received bFGF-, insulin-, and IGF-I-immobilized SGMs was significantly higher than that for the group that received insulin- and IGF-I-immobilized SGMs 4 weeks after the injection of microspheres. These results suggest that the system developed here is effective for the de novo formation of adipose tissue as it enables the induction of the two-step biological reaction by single injection.

Novel strategy for soft tissue augmentation based on transplantation of fragmented omentum and preadipocytes.

Current therapeutic procedures for soft tissue augmentation still lack the ability to induce rapidly formation of adipose tissue and its long-term stability, which is determined by rapid revascularization. The omentum is highly vascularized with microvascular endothelial cells (ECs) and is composed mainly of adipocytes that produce an enormously high level of vascular endothelial growth factor (VEGF). The aim of this study was to determine the potential usefulness of fragmented omentum tissues, with or without cotransplantation with preadipocytes, in soft tissue augmentation. Fragmented omentum tissues (approximately 500 mg) with or without preadipocytes (approximately 2.3 x 10(6)) isolated from epididymal adipose tissues were transplanted under the dorsal skin of Wistar rats by percutaneous injection and the tissues were left under the skin for up to 12 weeks. Regardless of cotransplantation with preadipocytes, the general morphological features of the transplanted tissue were as follows. The transplanted tissues, the weight loss of which was limited to 30-40%, contained viable adipocytes and some pseudocysts surrounded by fibrotic septa with minor inflammatory cell infiltration. High levels of triacylglycerol content, capillary density, and VEGF production were observed in transplanted tissues 12 weeks postoperation. Cotransplantation with preadipocytes enhanced adipose tissue formation significantly. These observations strongly indicate that transplantation of fragmented omentum tissues or cotransplantation with preadipocytes may be a promising therapeutic procedure for soft tissue augmentation.