Spatial and temporal control of laminin-332 and -511 expressions during hair morphogenesis.

Hair is one of the smallest organs, but has many important functions to mammals. Hair morphogenesis occurs through the reciprocal exchange of epithelial and mesenchymal signals. There are some reports about the expression of laminin-511 and -332 during hair morphogenesis, but are no reports of the chronological expression and function of laminin-511 and its counter regulator laminin-332 during hair morphogenesis. Our results of immunoblotting revealed that laminin-332 proteins were detected at stage 0 and downregulated during stage 1 to stage 2, and then recovered at stage 3. However, laminin α5 expression was constant throughout stages 0-3. According to the results of semi-quantitative RT-PCR, the mRNA expression of all laminin-332 subunits increased gradually from stage 0 to stage 2, while the mRNA expression of all laminin-511 subunits remained constant from stage 0 to stage 3. Our results suggest that the proper expression of laminin-332 and laminin-511 may regulate appropriate hair morphogenesis.

[A case of recurrent pancreatic cancer effectively responding to S-1 combined irinotecan third-line chemotherapy with PSK].

A case of recurrent pancreatic cancer effectively responded to S-1 and irinotecan combined with third-line chemotherapy (IRIS) with PSK. The patient was a 75-year-old female. In October 2007, a pancreatoduodenectomy was performed, followed with 6 courses of systemic adjuvant chemotherapy of gemcitabine (GEM). Three months after finishing the adjuvant chemotherapy, a recurrence of para-aortic lymph node metastasis was confirmed. We resumed the second-line chemotherapy of S-1/GEM (GS) with PSK. GS therapy was continued for about 3 years, until the recurrent lesion was found to have increased after 30 courses. Nevertheless, we continued up to 39 courses. In November 2011, we started third-line chemotherapy using S-1/irinotecan (IRIS) with PSK. The regimen was S-1 of 80 mg/body/day, continuously administered for day 1-14th, followed by a discontinuation for 2 weeks. CPT-11 100 mg/body/day was administered on day 1 and 15th; and PSK of 3 g/ body/day was continued, until it resulted in increased recurrent lesions. After the end of 4 courses, recurrent lesions started to respond partially. Currently, the patient is being treated as an outpatient. This case indicates that IRIS (S-1/CPT-11) is an effective therapy when pancreatic cancer fails to respond to GS therapy.

Protective effect of hochuekkito, a Kampo prescription, against ultraviolet B irradiation-induced skin damage in hairless mice.

A Kampo prescriptions, hochuekkito (HET) has been utilized for treating functional conditions such as general fatigue, compromised state and gastrointestinal motility disorder. Recently, HET has attracted the attention of dermatologists because of its clinically positive effects in atopic dermatitis (AD) treatment. To explain this positive effect of HET, we examined its protective ability against oxidative skin stress using a murine model. The dorsal region of 8-week-old male HR-1 hairless mice, which were raised on a HET (0%, 2% and 10%) mixed diet, was irradiated once with 70 mJ/cm(2) of ultraviolet (UV)-B light. After 4 days, transepidermal water loss (TEWL) and stratum corneum water content (SCWC), were determined as a measure of degree of skin dysfunction. To estimate the amount of active oxygen generated, the stratum corneum catalase activity (SCCA) and stratum corneum carbonylated protein (SCCP) content in the tape-stripped stratum corneum samples were measured. We also measured the H(2) O(2) scavenging ability of HET, and analyzed the changes in the expression levels of several inflammation and oxidative stress-related genes in the skin of HET-fed mice. In control mice, exposure to UV-B led to significant increases in TEWL and SCCP and significant decreases in SCWC and SCCA. These UV-B-induced changes were reduced in mice administrated HET, and the reduction was HET dose-dependent. Our results suggested that HET offered a protective effect against UV-B-induced skin damage. We also found that HET had relatively low ability to scavenge H(2) O(2) , and expression level of cyclooxygenase-2 mRNA decreased in HET-fed mouse.

Inducible nitric oxide synthase plays important roles in allergic reactions of pollinosis in mice sensitized with pollen allergy.

To elucidate the possible involvement of nitric oxide (NO) derived from inducible NO-synthase (iNOS) in the pathogenesis of patients with allergic rhinitis, we analyzed changes in the frequency of sneezing, plasma levels of NO metabolites, α-melanocyte-stimulating hormone (MSH) and immunoglobulin E and tracheal expression of IgA and mast cell tryptase in control and iNOS(-/-) mice. Eight-week-old control and iNOS(-/-) male C57BL/6j mice were sensitized with Cry j I antigen. After the last intranasal challenge of antigen, changes in the frequency of sneezing and plasma levels of IgE, α-MSH and NO metabolites and tracheal expression of iNOS, IgA and mast cell tryptase were analyzed by ELISA and immunohistochemistry using specific antibodies. The sensitization of mice with Cry j I antigen increased plasma levels of NO metabolites, α-MSH and IgE and tracheal expression of iNOS, IgA and mast cell tryptase in control not but in iNOS(-/-) mice. Administration of N(G)-nitro-L-arginine methyl ester strongly inhibited all these changes occurred in control mice. These results indicate that the symptom of pollinosis including sneezing is enhanced by iNOS derived NO through activation of α-MSH-receptor containing mast cells enriched with tryptase.

Mild exercise suppresses exacerbation of dermatitis by increasing cleavage of the ß-endorphin from proopiomelanocortin in NC/Nga mice.

This study investigated the mechanism by which the strength and weakness of exercise stress affects the skin symptoms of atopic dermatitis (AD). Specific pathogen-free (SPF) and conventional NC/Nga mice were used. Conventional mice, but not the SPF, spontaneously develop dermal symptoms similar to that of patients with AD. There were two types of stress, mild (20 m/min for 60 min) or strong exercise (25 m/min for 90 min), using a treadmill four times per day. The symptom of the conventional group were strongly exacerbated by strong exercise but ameliorated by mild exercise. The plasma concentrations of α-melanocyte stimulating hormone (α-MSH) and the expression of melanocortin receptor-1 in skin elevated after strong exercise but decreased after mild exercise. The plasma levels of ß-endorphin and the expression of µ-opioid receptor in skin were increased by mild exercise. In addition, the expression of prohormone convertase (PC) 1/3, PC2 and carboxypeptidase E (CPE) in pituitary gland were higher in the conventional group than in the SPF group. The level of PC2 was suppressed by mild exercise in the conventional groups, and elevated further by strong exercise. The level of PC1/3 becomes higher with the increase of the exercise load. On the other hand, the expression of the CPE was further increase by mild exercise but suppressed by strong exercise. These observations suggested that exercise-induced stress significantly affect the symptoms of AD in a pivotal manner depending on the levels of α-MSH and ß-endorphin, and the expression of pituitary PC2 and CPE.

Bullous pemphigoid IgG induces BP180 internalization via a macropinocytic pathway.

Bullous pemphigoid (BP) is an autoimmune blistering skin disease induced by pathogenic autoantibodies against a type II transmembrane protein (BP180, collagen type XVII, or BPAG2). In animal models, BP180 autoantibody-antigen interaction appears insufficient to develop blisters, but involvement of complement and neutrophils is required. However, cultured keratinocytes treated with BP-IgG exhibit a reduction in the adhesive strength and a loss of expression of BP180, suggesting that the autoantibodies directly affect epidermal cell-extracellular matrix integrity. In this study, we explored the consequences of two distinct epithelial cells treated with BP-IgG, particularly the fate of BP180. First, we followed the distribution of green fluorescent protein-tagged BP180 in an epithelial cell line, 804G, and normal human epidermal keratinocytes after autoantibody clustering. After BP-IgG treatment, the adhesive strength of the cells to their substrate was decreased, and BP180 was internalized in both cell types, together with the early endosomal antigen-1. By using various endocytosis inhibitors and a fluid-uptake assay, we demonstrated that BP-IgG-induced BP180 internalization is mediated via a macropinocytic pathway. Moreover, a macropinocytosis inhibitor rescued a BP-IgG-induced reduction in the adhesive strength of the cells from their substrate. The results of this study suggest that BP180 internalization induced by BP-IgG plays an important role in the initiation of disease pathogenesis.

UVB-induced epidermal pigmentation in mice eyes with no contact lens wear and non-UVB blocking and UVB blocking contact lens wear.

PURPOSE: Irradiation by ultraviolet (UV) B is known to increase the number of Dopa-positive melanocytes in the skin. This study examines the effectiveness of a contact lens for the defense of UVB eye irradiation-induced pigmentation. METHODS: A 2.5 kJ/m(2) dose of UVB radiation was delivered by a sunlamp to the eye of C57BL/6j male mice, and changes in the expression of Dopa-positive melanocytes in the epidermis and the plasma level of alpha-melanocyte-stimulating hormone (α-MSH) was analyzed. RESULTS: The degree of change in the Dopa-positive melanocytes expression was reduced by UVB blocking contact lens using mice given UVB irradiation to the eye. The plasma level of α-MSH increased in the C57BL/6j mice after irradiation to the eye, but there was no increase in the UVB blocking contact lens mice given UVB irradiation to the eye. Both the increase of the expression of Dopa-positive melanocytes and the plasma level of α-MSH were strongly suppressed by an alignment fitting UVB blocking contact lens and only a slightly suspended UVB blocking contact lens. In addition, these changes were successfully inhibited by a UVB blocking contact lens but not by a non-UVB blocking contact lens with a similar absorbance. CONCLUSION: These observations suggest that the UVB blocking contact lens inhibits the pigmentation of the epidermis in mice by suppressing of the α-MSH.

Intercellular pathway through hyaluronic acid in UVB-induced inflammation.

Ultraviolet B (UVB) radiation induces inflammation in the skin specifically at the site of exposure. We unexpectedly found that UVB-induced inflammation was not induced in gp91phox-depleted mice. To test whether gp91phox is directly involved in UVB-induced inflammation, neutrophil- and hyaluronic acid-depleted mice were also irradiated and examined for their response. Hyaluronic acid-depleted mice showed strongly inhibited UVB-induced inflammation, but the neutrophil-depleted mice did not exhibit any suppressed UVB-induced inflammation. To elucidate the pathway by which UVB irradiation induced inflammation, we examined the expression of nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3 (NLRP3) and caspase-1 in the mouse skin. An increase in the expression of NLRP3 and caspase-1 was seen following the UVB irradiation of C57BL mice; however, the UVB-irradiated gp91phox-knockout (gp91phox(-/-)) mice did not have this increase in expression. Furthermore, the plasma IL-1ß level increased after the UVB irradiation in C57BL mice, but there was no change in the gp91phox(-/-) mice. These results clearly indicate that nicotinamide adenine dinucleotide phosphate oxidase is activated by gp91phox, which is expressed on the surface in response to the increased expression of hyaluronic acid induced by UVB irradiation, and as result, the generation of reactive oxygen species (ROS) increases. This ROS activate NLRP3, and NLRP3 leads to the production of caspase-1, which subsequently increases IL-1ß, thereby finally inducing inflammation. It is thought that this system may play an important role in the damage and ageing of skin, and further studies are necessary to confirm these finding.

Effect of the smell of Seirogan, a wood creosote, on dermal and intestinal mucosal immunity and allergic inflammation.

Seirogan, a wood creosote, has been used as an antidiarrhetic drug in Asian countries including Japan for many years. This antidiarrhetic has recently been used as a sugar-coated pill because Seirogan has a strong smell. The strong smell of the uncoated form of Seirogan may modulate the defense systems of animals because the sense of smell is important for the detection of toxic metabolites in foods contaminated with pathogens. This study examined the effect of the sugar-coated and uncoated forms of this antidiarrhetic on the immunological response and inflammatory reactions in mice that had been sensitized with either fluorescein isothiocyanate or oxazolone. The sensitization of mice with either FITC or oxazolone markedly increased the plasma levels of tumor necrosis factor-α and mucosal IgA and elicited severe inflammation in the colon by a mechanism that could be suppressed by exposure of animals to the smell of uncoated Seirogan as effectively as the oral administration of the agent. Dermal inflammation in the FITC- and oxazolone-sensitized mice was also suppressed effectively either by the exposure to the smell or oral administration of the agent. Biochemical and histochemical analyses revealed that the elevated levels of plasma tumor necrosis factor-α and mucosal IgA were significantly decreased by exposure to the smell of uncoated Seirogan as well as by oral administration of the agent. Exposure of mice to the smell of Seirogan but not oral administration of the agent selectively increased plasma levels of adrenocorticotropic hormone and cortisol, particularly in the sensitized animals. These observations suggest that exposing the animals to the smell of Seirogan per se activated the hypothalamo-pituitary-adrenal axis and systemically modulated immunological reactions to suppress the allergic reactions.

Pediatric case report: clinical profile of a patient with PCWH with p.Q377X nonsense mutation in the SOX10 gene.

We report the case of a Japanese patient with PCWH, a neurological variant of Waardenburg type 4. Direct sequencing of the genomic DNA obtained from peripheral leukocytes revealed the p.Q377X nonsense mutation in the SOX10 gene. The patient had mottled hypopigmented macules on the trunk since birth; such macules have not been described previously. The so-called "white forelock", a triangular or diamond shaped leukoderma on the forehead, was absent. We also reviewed and summarized the outcomes of 23 patients with Waardenburg syndrome type 4, PCWH and Yemenite deaf-blind hypopigmentation syndrome, in which SOX10 mutations were identified. Among them, 17 cases were reported to have hypopigmented skin macule(s). The five patients who had the white forelock had PCWH with severe neurological complications. Paradoxically, two cases had hyperpigmented spots. Heterochromia of the iris was reported in four patients.

Two cases of steatocystoma simplex in infants.

Steatocystoma simplex is a rare benign cutaneous cyst that is thought to be a circumscribed malformation arising from the pilosebaceous duct junction. Reports describing steatocystoma simplex are scarce and many issues remain to be clarified, including frequency and age distribution. Steatocystoma simplex is generally considered to be a solitary, non-inherited tumor arising in adults; there have been no previous reports of steatocystoma simplex in infants. We encountered two cases of steatocystoma simplex arising on the head in infants less than 2 years old. Herein we discuss the clinical and pathological characteristics of these cases.

Lipoma with bone and cartilage components in the left axilla of a middle-aged woman.

UNLABELLED: Lipoma is the most common type of benign soft tissue tumor. However, lipoma containing bone or cartilage is very rare, especially a tumor containing both of these components. We encountered a 59-year-old woman who had a benign lipoma of the infraspinatus muscle that contained both osseous and chondral components. Histopathological examination revealed the presence of chondral and osseous elements mixed with mature adipose tissue. To the best of our knowledge, there has never been a report of benign lipoma containing osteochondral tissue at this location. Therefore, we report the first case of an axillary lipoma with bone and cartilage components. LEVEL OF EVIDENCE V: This journal requires that authors assign a level of evidence to each article.

Utility of Dermoscopy before and after Laser Irradiation in Port Wine Stains.

BACKGROUND: Port wine stains (PWSs) are commonly treated with pulsed dye laser (PDL) as a standard therapy. However, it is not easy to predict the minimal effective dose in the first treatment session. OBJECTIVE: The aim of this study was to assess whether dermoscopic findings before and after laser irradiation corresponded with the clinical improvement of PWS in patients undergoing PDL therapy. METHODS: Seven untreated PWSs in 6 patients (a male and 5 females), who presented to our hospital between May 2008 to January 2010, were assessed in this study. The mean age was 36.3 years, ranging from 14 to 57 years. A PDL with a wavelength of 585 nm and a spot size of 7 mm was used. Before and after test irradiation, patients underwent dermoscopy and clinical photography, and we assessed whether the dermoscopic findings corresponded with clinical improvement after 3 months. RESULTS: There were no obvious differences observed in the clinical photographs between each test level immediately after irradiation. However, dermoscopic photographs showed differences as the irradiated energy increased. These changes corresponded to the clinical improvement after 3 months. CONCLUSION: Our study indicates that the minimal effective fluence can be predicted by observing dermoscopic change immediately after irradiation. We think that examining the dermoscopic findings immediately after irradiation allows the laser surgeon to predict the minimal effective fluence and this prevents adverse effects of the skin.

A technique for auricular keloid core excision using a skin biopsy punch.

BACKGROUND: Keloids of the auricular region, resulting from ear piercing or external injury, are a common cosmetic problem. Surgical treatment followed by conservative management often is needed. The "hollowing out method for keloids of the auricle" retains the skin over the keloid lesion to minimize tension on the wound. This is considered to be the appropriate surgical treatment method, but skill is required to remove the keloid and retain the skin with a uniform thickness and appropriate form. MATERIAL AND METHODS: Four patients with auricle keloids were included in this study. All the patients were females between the ages of 18 and 29 years (average age, 24.8 years). Keloid core excision using a skin biopsy punch with the patient under local anesthesia was performed for all the patients. RESULTS: Use of a skin biopsy punch resulted in a shorter operating time without causing ear deformity. CONCLUSION: The authors report a technique for keloid core excision using a skin biopsy punch and believe it is a useful method LEVEL OF EVIDENCE V: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors at www.springer.com/00266.

Intense pulsed light therapy for aberrant Mongolian spots.

BACKGROUND: Aberrant Mongolian spots (AMS) distal to the lumbosacral region are thought to be more likely to persist than typical sacral Mongolian spots. So far, Q-switched ruby laser (QSRL) has been the treatment of choice for AMS. Intense pulsed light (IPL) is obtained from flashlamp devices that emit wavelengths between 515 and 1200 nm. IPL has documented efficacy for the treatment of irregular pigmentation, telangiectasia, rough skin texture, rhytids, hair removal, and vascular lesions, with several filters being available that can be used to block shorter wavelengths from the skin. As far as we could determine, there have been no clinical and histological studies on the treatment of AMS with IPL. Accordingly, the aim of this study was to assess the clinical and histological efficacy of IPL for AMS. METHODS: Seven patients (4 males and 3 females) presenting from September 2008 to July 2009 were assessed. Their mean age was 2.0 years, ranging from 0 to 7 years. The IPL device used in this study was a Natulight (Lumenis Ltd., Tokyo, Japan). Photographs were taken of all patients with a high-resolution digital camera at baseline and 6 months after treatment. Skin biopsy specimens were taken from 1 patient (case 4) before, immediately after, and 6 months after treatment. RESULTS: According to the 7 family members of the patients, the outcome of IPL was graded as follows: excellent improvement in 1 (14%), good improvement in 4 (57%), and slight improvement in 2 (29%). All families would have liked to continue IPL treatment. Evaluation of the effect of treatment by a physician was less favorable, with excellent improvement in 1 (14%), good improvement in 2 (29%), and slight improvement in 4 (57%). Histopathologic examination of the pigmented region revealed the typical features of a Mongolian spot in the hematoxylin-eosin stained section. Immediately after IPL, there were no changes in the dermis. At 6 months after treatment, however, the number of melanocytes in the middle and upper dermis was obviously decreased. CONCLUSIONS: IPL is an effective method for the treatment of AMS.

Dye laser photodynamic therapy for Bowen's disease in a patient with epidermodysplasia verruciformis.

Epidermodysplasia verruciformis (EV) is a rare heritable skin disease that results in unusual susceptibility to infection with specific types of human papillomavirus (HPV). Here we report a 53-year-old man with EV who developed Bowen's disease on his lower eyelid and the chest. Mutation analysis of EVER1 gene revealed homozygous splice acceptor site mutation (IVS8-2, A > T). In this patient, HPV3, HPV14, and HPV38 had been identified from the skin lesions. The Bowen's skin lesion on the left lower eye-lid was treated by photodynamic therapy (PDT) using 5-aminolevulinic acid (ALA) and pulsed dye laser (PDL). After two rounds of the PDT treatment, the skin lesion disappeared and a skin biopsy confirmed the efficacy of the treatment. This method was simple, less invasive than other treatments, and achieved a satisfactory cosmetic result.