Takafumi Kamiya

Basic fibroblast growth factor reduces scar formation in acute incisional wounds

In order to identify a means to reduce scar formation of the skin after incision, this study examined the effect of local administration of basic fibroblast growth factor (bFGF) in humans. bFGF was administered to a sutured wound immediately after an operation. The drug was injected once into the dermis of the margins of wounds using a 27G needle or rinsing after performing dermostitches. The lengths of the treated wounds varied from 1 to 32 cm, and the subjects were 2–86 years old. Sutured wounds after excision of skin tumors from the face, trunk, and limbs and sutured wounds such as those at the donor sites of full‐thickness skin grafts were treated with low‐dose bFGF injections (0.1 μg/cm wound; Group 2), high‐dose bFGF injections (1 μg/cm wound; Group 3), and rinsed with high‐dose bFGF (1 μg/cm wound; Group 4). No patient treated with bFGF had hypertrophic scars, while some patients had hypertrophic or very wide scars in the control group (Group 1), and the ratios of minimum scarring of Group 2 ( p<0.001), Group 3 ( p<0.0001), and Group 4 ( p<0.0001) were statistically significantly higher than those of Group 1. Postoperative administration of bFGF inhibited hypertrophic scarring and widening of remaining scars without any serious side effects.

The mechanisms underlying fibroblast apoptosis regulated by growth factors during wound healing

While investigating the mechanisms underlying cell death during wound healing processes, we uncovered the pro‐apoptotic effects of basic fibroblast growth factor (bFGF) on granulation tissue fibroblasts following pretreatment with transforming growth factor (TGF)‐β1 in vitro. bFGF induced caspse‐3 activation and apoptosis in TGF‐β1‐pretreated granulation tissue‐derived fibroblasts (GF‐1) following bFGF treatment for 48 and 96 h. In contrast, fibroblasts that had been treated in the same manner and that originated from the uninjured dermis did not display apoptosis, indicating that the mechanisms underlying apoptosis events in fibroblasts that originate from normal dermal and wound tissues differ. In this process, we also found that bFGF inhibited Akt phosphorylation at serine 473 and induced a rapid loss of phosphorylation of focal adhesion kinase (FAK) at tyrosine 397 in pretreated GF‐1 cells, an event that coincided with the dissociation of phosphorylated FAK from the focal adhesions. Therefore, inhibition of survival signals relayed via the disrupted focal adhesion structures and inactivated Akt following bFGF treatment may lead to apoptosis in GF‐1 cells pretreated with TGF‐β1. Pretreatment of GF‐1 with TGF‐β1 followed by the addition of bFGF resulted in significantly greater inhibition of phosphorylation of Akt and FAK compared to treatment with TGF‐β1 or bFGF alone. The combinatorial treatment also led to proteolysis of FAK and inhibition of FAK and Akt protein expression in GF‐1 cells. These findings demonstrated a significant role for the two cytokines in apoptosis of granulation tissue fibroblasts during wound healing. In vivo studies also confirmed a marked decline in phosphorylation and protein expression of Akt and FAK in bFGF‐injected skin wounds. These results led to the hypothesis that temporal activation of TGF‐β1 and bFGF at the injury site promotes apoptosis in granulation tissue fibroblasts, an event that is critical for the termination of proliferative granulation tissue formation. Copyright

Small cell variant of CD30+ primary cutaneous T-cell lymphoma with epidermotropism that completely regressed after incisional skin biopsy

rates of 25%, 45%, 83% and 100%, respectively. The probability of provocation was modelled using binary logistic regression in an equation with PLESI, current age, years since onset of PLE, buttock MED, gender, skin type and positive family history as predictor variables. All these variables, except the PLESI score, were not significant and were sequentially removed from the model. The resultant equation had the PLESI score as a single predictor variable (P 1⁄4 0Æ01) and was: P 1⁄4 exp ()2Æ93 + 0Æ062 · PLESI)/ [1 + exp ()2Æ93 + 0Æ062 · PLESI)]. This equation describes a sigmoid curve (see Fig. 1). In the model, the probability of successful provocation with PLESI scores of 25, 50 and 75 was 20%, 54% and 84%, respectively. The mean PLESI score of those not provoked and those provoked by three, two and one exposures was 46Æ5 (n 1⁄4 14), 51Æ3 (n 1⁄4 8), 69Æ3 (n 1⁄4 11) and 59Æ4 (n 1⁄4 3), respectively; this upward trend was significant (rs 1⁄4 0Æ42, P 1⁄4 0Æ01), despite the low last value. The use of SSR may not be the optimal method for provocation; for example, a source emitting wavelengths almost entirely in the UVA range might provoke a greater proportion of patients. Although further studies to assess the relationship between PLESI score and outcome of provocation are desirable, we have shown that the PLESI is associated with the likelihood of experimental provocation of the rash. This suggests that investigators wishing to provoke the eruption experimentally will benefit by selecting patients with high PLESI scores, and provides further validation of the PLESI. To interpret the results of PLE research fully, the severity of the disease in participating subjects needs to be known. Therefore, studies of the provocation, prevalence, pathogenesis and treatment of PLE would benefit by using the PLESI to consider disease severity in participating patients. Acknowledgments

Melanoma-Targeted Chemothermotherapy and In Situ Peptide Immunotherapy through HSP Production by Using Melanogenesis Substrate, NPrCAP, and Magnetite Nanoparticles

Exploitation of biological properties unique to cancer cells may provide a novel approach to overcome difficult challenges to the treatment of advanced melanoma. In order to develop melanoma-targeted chemothermoimmunotherapy, a melanogenesis substrate, N-propionyl-4-S-cysteaminylphenol (NPrCAP), sulfur-amine analogue of tyrosine, was conjugated with magnetite nanoparticles. NPrCAP was exploited from melanogenesis substrates, which are expected to be selectively incorporated into melanoma cells and produce highly reactive free radicals through reacting with tyrosinase, resulting in chemotherapeutic and immunotherapeutic effects by oxidative stress and apoptotic cell death. Magnetite nanoparticles were conjugated with NPrCAP to introduce thermotherapeutic and immunotherapeutic effects through nonapoptotic cell death and generation of heat shock protein (HSP) upon exposure to alternating magnetic field (AMF). During these therapeutic processes, NPrCAP was also expected to provide melanoma-targeted drug delivery system.

Clinical and epidemiological analysis in 149 cases of rhododendrol-induced leukoderma

Rhododendrol‐induced leukoderma is an acquired depigmentation that develops mainly at the contact site after repeated use of skin‐whitening cosmetics containing rhododendrol. In most cases, cessation of further depigmentation or occurrence of repigmentation is observed after discontinuing the use of cosmetics. However, some patients develop vitiligo vulgaris through the spread of depigmentation into the non‐exposed areas. Our study aims to investigate the patient‐specific factors that may affect the extent of depigmentation or repigmentation, as well as development of vitiligo vulgaris. The degree of depigmentation of the face, neck and hands where exposed to rhododendrol was scored using photographs over time. The relationships between depigmentation score at first visit/improvement rate of depigmentation score and patient demographics were evaluated and three important clinical observations were made. First, repigmentation of the face was superior compared with that of the hands and neck, suggesting a possible role for the migration and differentiation of melanocyte stem cells from hair follicles, as a mechanism of repigmentation. Second, the intensity of rhododendrol exposure did not contribute to differences in the severity of depigmentation. This suggested a possibility of underlying genetic susceptibility to melanocyte cytotoxicity or immune reaction. Third, depigmentation score at first visit and past history of atopic dermatitis were significantly high in patients who developed vitiligo vulgaris. This suggested that severe chemical damage of melanocytes by rhododendrol leads to a higher risk of developing vitiligo vulgaris through the possible involvement of an immune reaction. These clinical observations may help to further understand the pathogenesis of rhododendrol‐induced leukoderma.

Serum cytokeratin 19 fragment 21‐1 is a useful tumor marker for the assessment of extramammary Paget's disease

Cytokeratin 19 fragment 21‐1 (CYFRA 21‐1) has been used as a tumor marker for several malignancies. However, to date, no studies have assessed whether CYFRA 21‐1 could be a useful marker for extramammary Pagets disease (EMPD). The present study aimed to evaluate the significance of CYFRA 21‐1 as a serum tumor marker for EMPD progression. Concentrations of serum CYFRA 21‐1 and carcinoembryonic antigen (CEA) in 13 cases of EMPD were measured prior to undergoing treatment at Sapporo Medical University Hospital from January 2014 to May 2016. Four of the 13 patients had lymph node metastases at diagnosis, but none had distant metastases. Immunohistochemistry indicated that all 13 primary tumors and four metastatic tumors in lymph nodes were positive for cytokeratin 19. Although none of the 13 patients showed high serum CEA levels, six patients (46.2%) had elevated serum CYFRA 21‐1. Furthermore, CYFRA 21‐1 was reduced in association with post‐treatment tumor reduction in all six patients. Among these six patients, four developed recurrence and metastasis during the follow‐up period. CYFRA 21‐1 was re‐elevated in all four of these patients; however, serum CEA was elevated only in the patient with distant metastasis. These results suggest that CYFRA 21‐1 is more sensitive compared with CEA, and can be useful as a tumor marker for evaluating tumor progression and treatment efficacy in patients with EMPD.

Nagashima-type palmoplantar keratosis caused by compound heterozygous mutations in SERPINB7

Nagashima-type palmoplantar keratosis (NPPK, MIM 615598) is an autosomal recessive disorder, first described by Nagashima [1-4]. It is characterized by diffuse mild palmoplantar hyperkeratosis that extends onto the dorsum of hands and feet and the Achilles tendon areas. Elbows and knees are also involved and palmoplantar hyperhidrosis is common. Recently, causative loss-of-function mutations in the SERPINB7 gene were identified [4-6]. Here we report a case of a patient with compound heterozygous [...]

Increased Caspase-2 Activity is Associated with Induction of Apoptosis in IFN-β Sensitive Melanoma Cell Lines

Interferon (IFN) is believed to be one of the most effective anti-melanoma agents. Specifically, IFN-beta has the ability to induce apoptosis of melanoma cells. Induction of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has also been suggested to have a critical role in IFN-beta-induced apoptosis. To characterize the signaling pathway involved in IFN-beta-induced apoptosis, we analyzed the biological effects of IFN-beta on the cell death and caspase activation of melanoma cells. IFN-sensitive cell lines, MM418, SK-mel-23, and SK-mel-118, showed increased apoptotic populations correlated with the activation of caspase-2 and caspase-3 by IFN-beta. IFN-beta-induced apoptosis was significantly suppressed by inhibitors for caspase-2 or caspase-3, but not by inhibitors for caspase-8 or caspase-9 in these cell lines. TRAIL expression was observed in IFN-beta-treated cells of SK-mel-23 and SK-mel-118, but not in those cells of MM418, which showed massive IFN-beta-induced apoptosis and resistance to exogenous TRAIL-mediated apoptosis. G361 was resistant to IFN-beta-induced apoptosis but sensitive to exogenous TRAIL-mediated apoptosis. Furthermore, IFN-beta pretreatment significantly increased the sensitivity against exogenous TRAIL-mediated apoptosis and activation of caspase-2 in G361. These results suggested that caspase-2 activation is commonly associated with induction of IFN-beta-induced apoptosis in IFN-beta-sensitive melanoma cells.

De novo follicular regeneration of the skin by wingless int 3 and bone morphogenetic protein 2 genes introduced into dermal fibroblasts and fibroblast growth factor-2 protein

In this study, we regenerated skin and its appendages by transplanting cultured normal dermal fibroblasts, into which morphogen genes had been introduced. We cultured normal dermal fibroblasts obtained from Fisher 344 rats on the surface of hydroxyapatite beads, and then adsorbed them onto the surface of a collagen sponge, which was transplanted into a full‐thickness skin defect prepared on the backs of rats. Before transplantation, genes were introduced into the dermal fibroblasts via adenovirus vector (ad)‐bone morphogenetic protein 2 and ad‐wingless int 3 genes in addition to fibroblast growth factor‐2 protein. By Week 4, the appearance of follicle germs or primitive hair germs was observed only in the ad‐bone morphogenetic protein 2+ad‐wingless int 3 combined with the fibroblast growth factor‐2 protein group. By Week 16, in that same group, hair follicles having mature pilosebaceous systems with equally spaced localization had formed in the ulcer wound.

Mucous membrane pemphigoid accompanied by ovarian cancer: A case with autoantibodies solely against γ2-subunit of laminin-332

phigus is greater than the general population, there may be a possibility of simple coincidence of IgA pemphigus and MM in these four patients. Further studies on the pathological role of MM in patients with IgA pemphigus are needed to elucidate the pathogenesis of this association. Jae Yong SUNG, Sang Eun LEE, Soo-Chan KIM Department of Dermatology and Cutaneous Biology Research Institute, Yonsei University College of Medicine, Seoul, and Department of Dermatology, CHA Bundang Medical Center, CHA University, Seongnam, Korea