Journal of Dermatological Science - Articles in Press

Differential expression of T cell immunoglobulin- and mucin-domain-containing molecule-3 (TIM-3) according to activity of Behçet's disease - Corrected Proof

Joong Sun Lee, Mi Jin Park, Sun Park, Eun-So Lee — 2012-02-16

Behçet's disease (BD) is a systemic chronic inflammatory disorder characterized by recurrent oral aphtha, genital ulcers, uveitis, and skin lesions. The etiology and pathogenesis of BD are unknown; however, many studies show a predominant T helper (TH1)-mediated immune response in BD . Recently, a novel cell surface protein, T cell immunoglobulin and mucin domain (TIM), was identified . Among three genes (TIM-1, TIM-3 and TIM-4) in humans, TIM-3 was found to be specifically expressed by differentiated TH1 cells . Later TIM-3 expression was demonstrated in other cell populations such as Th17, CD8+ cells, NK cells, NCT cells and monocytes . The role of Tim-3 in autoimmune diseases and inflammatory diseases has been accumulated. In vivo blockade of Tim-3 enhanced the onset in an adoptive transfer model of autoimmune diabetes . In multiple sclerosis, preferential TIM-3 expression on TH1 cells is reported. Considering that TH1 immune responses play an important role in the pathogenesis of BD, we wanted to examine the expression of TIM-3 in BD patients and to determine whether TIM-3 expression is correlated with disease activity.

Eleven novel mutations of the ADAR1 gene in dyschromatosis symmetrica hereditaria - Corrected Proof

2012-02-16

Dyschromatosis symmetrica hereditaria (DSH: MIM#127400) shows an autosomal dominant pattern of inheritance with high penetrance, which is characterized by hyperpigmented and hypopigmented macules on the face and dorsal aspects of the extremities. The phenotypes commonly appear in infancy or early childhood . In Asia, the condition occurs predominantly among Japanese and Chinese individuals. Previous study have clarified that a heterozygous mutation of the adenosine deaminase acting on RNA1 (ADAR1, formerly DSRAD) gene causes DSH in Japanese DSH families . The human ADAR1 spans 30kb and contains 15 exons. It encodes RNA-specific adenosine deaminase composed of 1,226 amino acid residues. The enzyme has two Z-alpha domains, three dsRNA binding domains and the deaminase domain . To date, more than 90 mutations have been reported from East Asian countries so far , which confirmed that the ADAR1 gene is responsible for DSH not only in Japanese but also in other ethnic groups. ADAR1 protein catalyzes the deamination of adenosine to inosine in double-stranded RNA substrates , which results in the creation of alternative splicing sites or alternations of the codon and thus leads to functional changes in the target protein. The ADAR1 gene is expressed ubiquitously, but the target RNA(s) in the skin as well as the mechanisms by which mutations in ADAR1 gene cause DSH still remain unknown. DSH patients are heterozygous for the ADAR1 gene mutation that is inherited as a dominant trait. Unlike patients with DSH, mice that are heterozygous for Adar1 gene deletion did not manifest any skin symptom. Homozygous Adar1 gene deletion was embryonic lethal. Recently epidermis-specific Adar1 gene knockout mouse model was generated . Histologically these mice showed massive necrosis of epidermis in FAB background mice, and thickened keratinocyte and stratum corneum in B6 background mice. Therefore, Adar1 was thought to be an essential molecule for skin integrity. In this study, we describe mutation analyses of Japanese families with DSH. The mutational analysis of the ADAR1 gene was performed as previously described . Informed consent and blood samples of patients were obtained under protocols approved by the Ethics Committee of Yamagata University School of Medicine.

Rare hereditary autoinflammatory disorders: Towards an understanding of critical in vivo inflammatory pathways - Corrected Proof

Nobuo Kanazawa — 2012-02-15

Abstract: Hereditary autoinflammatory syndromes are monogenic disorders with an inborn error of innate immunity, and include periodic fever syndromes such as familial Mediterranean fever (FMF), tumor necrosis factor receptor-associated periodic syndrome and cryopyrin-associated periodic syndromes (CAPS), pyogenic diseases such as pyogenic arthritis, pyoderma gangrenosum and acne syndrome (PAPAS), and granulomatous diseases such as Blau syndrome. By identifying the genetic abnormalities and subsequent analyses of the molecular mechanisms underlying these disorders, several critical in vivo pathways for inflammatory processes have been discovered. In this review, three categories of autoinflammatory disorders are discussed: inflammasomopathies, receptor antagonist deficiencies and proteasome disability syndromes. Inflammasomopathies are diseases with dysregulated NLRP3 inflammasome activation, and include CAPS with NLRP3, FMF with MEFV, and PAPAS with PSTPIP1 mutations. Analyses of these diseases have clarified some critical pathways regulating NLRP3 inflammasome signaling. Receptor antagonist deficiencies include the newly defined deficiency for interleukin-1 receptor antagonist resulting in sterile multifocal osteomyelitis with periostosis and pustulosis, and deficiency for interleukin-36 receptor antagonist resulting in generalized pustular psoriasis. The identification of these genetic abnormalities has revealed a critical role for receptor antagonists of IL-1 family cytokines in regulating neutrophil activation/recruitment. Finally, proteasome disability syndromes with PSMB8 mutations include Nakajo-Nishimura syndrome and related disorders distributed globally. Analyses of these diseases have unexpectedly shown a critical role of the ubiquitin–proteasome system in the regulation or homeostasis of inflammation/metabolism. Since there still remain a number of predicted but undefined hereditary autoinflammatory syndromes, further clinical and genetic approaches are required to discover novel in vivo critical inflammatory pathways.

Interaction of plectin and intermediate filaments - Corrected Proof

2012-02-14

Abstract: Background: Plectin, a member of the plakin family proteins, is a high molecular weight protein that is ubiquitously expressed. It acts as a cytolinker for the three major components of the cyotoskeleton, namely actin microfilaments, microtubules and intermediate filaments.Objective: The aim of our experiments was to identify new binding sites for intermediate filaments on plectin and to specify these sites.Methods: We introduced truncated forms of plectin into several cell lines and observe interaction between plectin and intermediate filaments.Results: We found that a linker region in the COOH-terminal end of plectin was required for the association of the protein with intermediate filaments. In addition, we also demonstrated that a serine residue at position 4645 of plectin may have a role on binding of plectin to intermediate filaments.Conclusion: A linker region in the COOH-terminal end and serine residue at position 4645 may be important for the binding of plectin to intermediate filaments.

Mcl-1 determines the imiquimod-induced apoptosis but not imiquimod-induced autophagy in skin cancer cells - Corrected Proof

2012-02-06

Abstract: Background: Imiquimod had been shown to induce apoptosis and autophagy in several skin cancer cells, especially basal cell carcinoma (BCC) cells.Objective: We evaluate the molecular mechanisms of imiquimod-induced apoptosis and autophagy in skin cancer cell lines.Methods: The Mcl-1, Bcl-2 and Bcl-xL proteins were determined by immunoblotting. The Mcl-1 mRNA level was examined by RT-PCR and real-time PCR. The mechanisms of imiquimod-induced decrease in Mcl-1 protein were evaluated by addition of cycloheximide, MG132 proteasome inhibitor or pan-caspase inhibitor. The phosphorylation of eIF4E, 4E-BP1 and eEF2 in imiquimod treated cells were examined by immunoblotting. The imiquimod-induced apoptosis and autophagy were evaluated in Mcl-1-overexpressing cells by XTT test, mitochondrial membrane potential measurement, DNA content assay, LC3 immunoblotting, EGFP-LC3 puncta formation and quantification of acidic vesicular organelle with acridine orange staining.Results: The decrease in the Mcl-1 protein level was faster and stronger than the decrease in Bcl-2 and Bcl-xL in imiquimod-treated skin cancer cells. The imiquimod-induced decrease in Mcl-1 protein was not caused by blocked transcription or the promotion of degradation but was associated with inactivation of translation factors in BCC cells. The Mcl-1-overexpressing BCC cells were more resistant to intrinsic cellular apoptosis than control BCC cells during imiquimod treatment. Mcl-1 overexpression in BCC cells resulted in the basal activation of autophagy but did not modulate imiquimod-induced autophagy or rescue imiquimod-induced autophagic cell death in BCC cells.Conclusions: Imiquimod may rapidly downregulate Mcl-1 protein levels by inhibiting translation in skin cancer cells. Mcl-1 may act to protect against apoptosis but not autophagy and autophagic cell death during imiquimod treatment in skin cancer cells.

Spectroscopic evidence of abnormal structure of psoriasis vulgaris stratum corneum - Corrected Proof

Kouichi Nakagawa, Satoko Minakawa, Daisuke Sawamura — 2012-02-06

EPR (electron paramagnetic resonance) is useful for elucidating structural aspects of stratum corneum (SC) . Non-invasive spectroscopic characterization of the outermost layer of the SC is an important subject in dermatology and cosmetology. The SC, a heterogeneous structure composed of corneocytes embedded in the intercellular lamellar lipid bilayer, acts as the main epidermal barrier against chemicals, oxidative stress, and other invasive environmental factors, as well as regulating TEWL (transepidermal water loss) to prevent dehydration of viable cells underneath the SC. The measurement of TEWL is the most common method to estimate the state of the skin. However, the TEWL does not provide structural information about the lipid in the SC.

Inflammatory response of human keratinocytes triggered by Sporothrix schenckii via toll-like receptor 2 and 4 - Corrected Proof

Min Li, Qing Chen, Jun Sun, Yongnian Shen, Weida Liu — 2012-02-06

Sporotrichosis is the most common cutaneous and subcutaneous mycosis caused by the dimorphic fungus Sporothrix schenckii of which conidia and/or hyphae could convert into yeast form at the site of infection after traumatic inoculation . Histopathological findings of early cutaneous sporotrichosis showed an infiltration of neutrophils and other inflammatory cells within the epidermis and dermis . Recently Sassá et al. reported that activation of macrophage TLR4 played an important role in host defense against S. schenckii . We sought to clarify whether human keratinocytes play immunological roles in the inflammatory response during the early phase of S. schenckii infection through TLR2 and TLR4.

Migration of human melanocytes into keratinocyte monolayers in vitro - Corrected Proof

Dheshnie Keswell, Lester M. Davids, Susan H. Kidson — 2012-02-06

The repigmentation of adult skin after wound healing, or loss of melanocytes (e.g. vitiligo), poses a particular and unique challenge to the invading population of melanocytes. The melanocytes, or their precursors, resident in adjacent skin or in nearby niches, must be activated to proliferate, and then negotiate their way through a tightly held epidermal barrier. The mechanisms whereby melanocytes negotiate their way through the epidermis to achieve this repigmentation have not been described in any detail and may occur trans-epidermally, and/or along the basement membrane and/or in the dermis. While there have been many studies on migratory cells during embryonic development and in particular physiological processes , these situations differ from the adult skin epidermal environment. To migrate through the tight spaces between keratinocytes, either junctions must be broken and spaces thus created, or alternatively, quite extreme cellular contortions would need to occur to allow the cell to move through the narrow paracellular spaces. Further knowledge of these processes will certainly contribute towards elucidating the mechanisms of repigmentation in pigmentary disorders.

Epidermogenesis in a skin wound deep through the basement membrane contributes to scar formation - Corrected Proof

Lujun Yang, Koji Hashimoto, Yuji Shirakata — 2012-02-03

In normal human skin, the basement membrane (BM) lies between the epidermis and dermis, connecting the two compartments tightly and keeping them from direct contact. During skin wound healing, fibroblasts from adjacent intact dermis are activated and migrate to fibrin clots to form granulation tissue, onto which activated keratinocytes at the wound margin migrate, stratify, and differentiate to form a new integument; however, a well-developed BM is not observed until a few months later . During this period, homeostasis between the epidermal and dermal compartments is interrupted and abnormal epidermal–dermal events can ensue.

The dimensions and characteristics of the subepidermal nerve plexus in human skin – Terminal Schwann cells constitute a substantial cell population within the superficial dermis - Corrected Proof

Christina M. Reinisch, Erwin Tschachler — 2012-02-03

Abstract: Background: The skin constitutes the largest sensorial organ. Its nervous system consists of different types of afferent nerve fibers which spread out immediately beneath the skin surface to sense temperature, touch and pain.Objective: Our aim was to investigate the dimension and topographic relationship of the different nerve fibers of the subepidermal nerve plexus in human hairy skin and to analyze numbers and marker expression of terminal Schwann cells.Methods: Nerve fibers and Schwann cells were investigated on dermal sheet preparations and thick sections of skin from various body regions of 10 individuals.Results: The dimension of subepidermal nerve fibers varied between different body sites with highest values in chest skin (100±18mm/mm2) and lowest in posterior forearm skin (53±10mm/mm2). The majority of fibers (85.79%) were unmyelinated, thus representing C-fibers, of which 7.84% were peptidergic. Neurofilament-positive fibers (A-fibers) accounted for 14.21% and fibers positive for both neurofilament and myelin (Aβ-fibers) for only 0.18%. The number of Schwann cells varied in accordance with nerve fiber length from 453±108 on chest skin to 184±58/mm2 in skin of the posterior forearm. Terminal Schwann cells showed a marker profile comparable to Schwann cells in peripheral nerves with the notable exception of expression of NGFr, NCAM, L1CAM and CD146 on myelinating Schwann cells in the dermis but not in peripheral nerves.Conclusion: Our data show that terminal Schwann cells constitute a substantial cell population within the papillary dermis and that both nerve fiber length and Schwann cell numbers vary considerably between different body sites.

TNF-α increases lipogenesis via JNK and PI3K/Akt pathways in SZ95 human sebocytes - Corrected Proof

2012-02-03

Abstract: Background: Tumor necrosis factor-alpha (TNF-α) is an important pathophysiologic factor involved in the development of acne. However, its role is unclear.Objective: To explore the lipogenic effect by TNF-α and possible molecular mechanisms in sebocyte.Methods: Using SZ95 human sebocytes, lipid formation by TNF-α was assessed by Oil Red O, Nile Red staining and thin layer chromatography (TLC). Expression of lipogenic genes and activation of mitogen-activated protein kinase as well as Akt were examined by real-time polymerase chain reaction and/or Western blot analysis. Activation of peroxisome proliferator-activated receptor (PPAR) was evaluated by luciferase assay using PPAR response element containing reporter plasmids. Involvement of c-Jun N-terminal kinase (JNK) and Akt in TNF-α-induced lipogenesis was investigated by molecule specific small interfering RNA and inhibitors.Results: TNF-α treatment significantly increased formation of lipid droplets in accordance with up-regulated expression of FAS and activation of SREBP-1, but not PPARs. Suppression of phosphorylated JNK by the JNK inhibitor SP600125 greatly diminished TNF-α-induced expression of FAS and SREBP-1. TNF-α could not induce both expression of lipogenic proteins and lipid synthesis when Akt expression was attenuated with siRNA.Conclusions: TNF-α induces lipogenesis in SZ95 human sebocytes through the JNK and phosphoinositide-3-kinase/Akt pathways. These results will be valuable in developing therapeutic strategies for control of seborrhea and acne.

Possible role of infrared or heat in sun-induced changes of dermis of human skin in vivo - Corrected Proof

Hyun-Sun Yoon, Yeon Kyung Kim, Mary Matsui, Jin Ho Chung — 2012-02-03

The signs of photoaging in human skin have been attributed almost exclusively to the ultraviolet radiation (UVR) which represents just 6.8% of solar radiation, compared to the infrared and visible radiation which account for 54.3% and 38.9%, respectively, of the incident solar energy . Little is known about the biologic effects of non-UVR components of solar radiation on human skin. Recent studies suggest that infrared radiation induces extracellular matrix (ECM) changes including increased matrix metalloproteinases and decreased procollagen expression , which manifest over time as wrinkles . Additionally, increased skin temperature by infrared radiation as well as heat alone can cause ECM changes similar to UVR.

Heparin serves as a natural stimulant of the inflammasome and exacerbates the symptoms of tumor necrosis factor receptor-associated periodic syndrome (TRAPS) - Corrected Proof

Shun Ohmori, Ryosuke Hino, Motonobu Nakamura, Yoshiki Tokura — 2012-01-31

Tumor necrosis factor receptor-associated periodic syndrome (TRAPS) is a rare autosomal dominant disorder characterized by recurrent episodes of fever, myalgia, abdominal pain, conjunctivitis and skin eruptions, which occur spontaneously or after some triggers. This syndrome is associated with missense mutations in TNFRSF1A, the gene encoding the 55kDa type 1 tumor necrosis factor receptor (TNFR1) . TRAPS-associated mutant TNFR1 accumulates intracellularly and does not act as a conventional TNF receptor . Although the pathogenesis of TRAPS has not been fully elucidated, the accumulation of mutant TNFR1 causes spontaneous mitogen-activated protein kinase (MAPK) activation or nuclear factor κB (NF-κB) activation . In TRAPS patients, therefore, even minor triggers that stimulate innate immune system can lead to inflammatory cytokine production, such as interleukin-1β (IL-1β), IL-6 or TNFα .

Modulation of skin oxidative stress and inflammatory markers by environmental stressors. Differences between young and old - Corrected Proof

2012-01-25

Beside ultraviolet radiation (UV), the skin is exposed to other stressors such as cigarette smoke (CS) and ozone (O3). O3 is not a radical species per se and its toxic effect is mediated by the production of a cascade of bioactive molecules (i.e. hydroxyl radical and aldehydes). CS contains more than 4300 different chemicals, most of which are classified as carcinogens for several tissues including the skin. Both, O3 and CS have been shown to induce oxidative stress and to deprive tissues of their antioxidants defense . There is increasing evidence that the susceptibility to air toxicants effect varies from one individual to another and those who suffer more from exposure to air pollution are the elderly . We have previously documented that O3 and CS modulate MMP's activity and the responses are affected by age . To further explore the skin's response to oxidants, young (8-week-old) and aged (18-month old) SKH1 mice were exposed to O3 (6h/day at 0.25ppm) or CS (6h/day) for 4 days as previously described . Oxidative stress and inflammation markers were analyzed by Western blot in skin tissue homogenate. All statistical analyses were performed using one-way analysis of variance (ANOVA).

The suppressive effects of ultraviolet radiation on immunity in the skin and internal organs: Implications for autoimmunity - Corrected Proof

Gary M. Halliday, Diona L. Damian, Sabita Rana, Scott N. Byrne — 2012-01-25

Abstract: Low doses of sunlight that can be received during normal daily activities suppress immunity in humans. Both ultraviolet (UV) B (290–320nm) and UVA (320–400nm) are immunosuppressive. The wavelength dependence in humans shows distinct non-overlapping immunosuppressive peaks of solar effectiveness centred at 310nm UVB and 370nm UVA. In murine models of systemic immunosuppression low dose UV inhibits expansion of effector T cells in skin-draining lymph nodes, and retention of dermal effector memory CD8T cells at sites of antigen challenge. In addition to suppressing skin immunity, UV inhibits immunity in internal organs, including activation of CD8 T cells and cytotoxic T cell activity in the spleen, and memory T cell activation in the spleen and bone marrow. Neither of the chromophores responsible for UV suppression of skin immunity, DNA damage and urocanic acid, nor reactive oxygen species are involved in regulation of CD8 T cells in internal organs. Thus UVB impedes the activation and cytotoxicity of antigen-specific T cells in internal organs by mechanisms independent of suppression of skin immunity. These deleterious effects of low dose UV on skin immunity are likely to contribute to skin cancer, however UV suppression of immunity in internal organs may protect from autoimmunity. Epidemiological evidence suggests that sunlight protects from some autoimmune diseases directed towards internal organs. As UV suppression of skin and internal organ immunity appear to occur via different mechanisms, it may be possible to protect skin immunity and therefore reduce skin cancer incidence without preventing UV from reducing autoimmunity in internal organs.

Spatial and temporal analysis of skin glycation by the use of multiphoton microscopy and spectroscopy - Corrected Proof

Ara A. Ghazaryan, Po-Sheng Hu, Shean-Jen Chen, Hsin-Yuan Tan, Chen-Yuan Dong — 2012-01-25

Abstract: Background: Tissue glycation, the main cause of many diabetes-related complications, results in the accumulation of advanced glycation endproducts (AGE).Objectives: These AGEs are endogenous fluorophores that can serve as a viable pathological indicator for disease diagnostics. Here we explore the capabilities of multiphoton microscopy to non-invasively localize and quantify the skin glycation.Methods: In our study, multiphoton microscopy and spectroscopy were used to investigate glycation events-induced changes in the intensities of autofluorescence and second harmonic generation on ex vivo human skin.Results: Temporal and spatial dependence of degrees of glycation of the epidermis, collagen and elastin fibers of dermis were evaluated for their relevance to the changes in amplitudes of autofluorescence signals. We found that glycation drastically and linearly increases multiphoton autofluorescence intensity of epidermis and dermal collagen whereas changes in dermal elastin are moderate. We also found decrease in the level of second harmonic generation signal.Conclusion: Our study suggests that due to intrinsically weak autofluorescence the dermal collagen is the most sensitive skin tissue to be used for detecting changes in tissue glycation.

Hydrogen peroxide and monoethanolamine are the key causative ingredients for hair dye-induced dermatitis and hair loss - Corrected Proof

2012-01-24

Summary: Background: Hair dyes are being commonly used to change the color of hair for cosmetic reason. However, concern is growing over the dermatitis and subsequent hair loss associated with the repeated use of hair dye products, yet the causative ingredients have not been elucidated.Objective: Here we investigated hair dye-induced dermatitis and hair loss using in vivo mouse model to uncover the causative ingredients.Methods: Commercially available hair dye products or combination of the ingredients of hair dye product were applied topically for 3 days on the dorsum of the female C57BL/6 mice and, dermatitis and hair loss were examined.Results: The mice treated with hair dye products exhibited unequivocal signs of hair loss and dermatitis. To find out causative ingredients, combinations of the representative components of hair dye including reducing agents, the mixture of dye and monoethanolamine (MEA), ammonia, and hydrogen peroxide (H2O2) were applied and thereafter, hair loss and dermatitis were evaluated. All the groups treated with the combinations containing H2O2 and neutralized dye mixture manifested hair loss and dermatitis. Subsequent experiments revealed that H2O2 and MEA synergistically induced hair loss and dermatitis. Histological examination showed that oxidative stress may be the mechanism underlying hair-dye induced dermatitis. Consistently, H2O2 and MEA synergistically induced oxidative stress and cytotoxicity in human keratinocytes.Conclusion: These results suggest that H2O2 and MEA may be the key causative ingredients for hair dye-associated dermatitis and hair loss.

Molecular characterization of two novel KIT mutations in patients with piebaldism - Corrected Proof

Bartosz Wasag, Magdalena Chmara, Eric Legius — 2012-01-20

Piebaldism (OMIM 172800) is a rare autosomal dominant disorder characterized by congenital depigmented patches of the mid-forehead, chest, abdomen and extremities . The disorder is caused by inactivating mutations or deletions of the KIT gene on chromosome 4q12 or the SLUG gene on chromosome 8q11. Here we report two new KIT mutations, a missense mutation p.T619A and an in frame p.T437dup duplication, associated with piebaldism and we show that both non-protein truncating mutations have an inactivating effect on the KIT protein and are pathogenic.

Knocking-in the R142C mutation in transglutaminase 1 disrupts the stratum corneum barrier and postnatal survival of mice - Corrected Proof

2012-01-18

Abstract: Background: Mutations in the gene encoding transglutaminase 1 (TG1) are responsible for various types of autosomal recessive congenital ichthyosis (ARCI), such as lamellar ichthyosis (LI), congenital ichthyosiform erythroderma (CIE) and some minor variants of ARCI. A point mutation of R143C in the β-sandwich domain of TG1 has been often identified in patients with LI or CIE.Objective: To elucidate the effect of that point mutation on skin barrier structures and functions, we generated mice with a point mutation of R142C, which corresponds to the R143C mutation in human TG1.Methods: A mouse line with the R142C point mutation in TG1 was established using a gene targeting technique and the Cre-loxP system. The skin phenotypes were analyzed in homozygous mutant Tgm1R142C/R142C mice.Results: In the skin of Tgm1R142C/R142C mice, expression of the mutant transcripts was comparable with wild-type or Tgm1+/R142C mice. However, the amount of mutated protein in the skin was markedly decreased in Tgm1R142C/R142C mice, and the TG1 activity of Tgm1R142C/R142C keratinocytes was almost lost. Tgm1R142C/R142C mice exhibited morphological and functional skin barrier defects and neonatal lethality. The stratum corneum of those mice lacked cornified envelopes, and loricrin, the major structural component, failed to assemble at the corneocyte cell periphery. Tgm1R142C/R142C mice showed a marked increase in transepidermal water loss and their skin was easily permeable to toluidine blue dye. The intercellular lipid lamellar structures of the stratum corneum were irregular and the 13-nm periodic X-ray diffractions from the stratum corneum lipid molecules were lost in vivo.Conclusion: From these results, we suggest that the R142C mutation of TG1 reduces the enzyme stability which is indispensable for development of the stratum corneum and skin barrier function and for postnatal survival of mice.

The retinoic acid-induced up-regulation of insulin-like growth factor 1 and 2 is associated with prolidase-dependent collagen synthesis in UVA-irradiated human dermal equivalents - Corrected Proof

2012-01-16

Abstract: Background: Ultraviolet (UV) A irradiation causes the degeneration of extracellular matrix in the skin dermis, mainly due to disrupted collagen homeostasis, resulting in the photo-aging of human skin. All-trans retinoic acid (ATRA) improves photo-aged human skin in vivo.Objectives: Although the effects of ATRA on collagen synthesis and MMP regulation are well known, the effects of ATRA on other collagen homeostasis-associated genes have not been elucidated. This study was aimed to study the factors that are pharmacologically associated with the effect of ATRA on collagen homeostasis.Methods: The gene transcription profile of collagen homeostasis-associated genes was systematically evaluated in three-dimensional human dermal equivalents (HDEs) following UVA-irradiation and/or ATRA treatment.Results: In addition to the expected changes in MMPs and collagen synthesis in HDEs in response to ATRA, prolidase, an important enzyme in the recycling of proline and hydroxyproline from degraded collagen molecules, was significantly decreased by UVA irradiation, and its down-regulation was antagonized by ATRA. Transfection with a prolidase-specific siRNA led to a significant decrease in procollagen synthesis in human fibroblasts. ATRA inhibited the UVA irradiation-induced decrease in prolidase activity through an insulin-like growth factor (IGF) receptor signaling pathway in HDEs. ARTA increased IGF1 and IGF2 production in HDEs, and neutralizing IGFs with anti-IGF antibodies abolished the effect of ATRA on proliase activity.Conclusions: These data demonstrate that ATRA regulates prolidase activity in HDEs via IGF receptor signaling, suggesting one of the pharmacological mechanisms by which improves photo-aged human skin.

The predominant drug-specific T-cell population may switch from cytotoxic T cells to regulatory T cells during the course of anticonvulsant-induced hypersensitivity - Corrected Proof

Takaaki Hanafusa, Hiroaki Azukizawa, Sayaka Matsumura, Ichiro Katayama — 2012-01-09

Abstract: Background: Delayed hypersensitivity is responsible for severe cutaneous adverse drug reactions (cADRs), especially in Stevens-Johnson syndrome (SJS), toxic epidermal necrolysis, and drug-induced hypersensitivity syndrome (DIHS) (also known as drug rash with eosinophilia and systemic symptoms [DRESS] syndrome). The drug-induced lymphocyte stimulation test (DLST), or lymphocyte transformation test (LTT), is used to identify the culprit drug in severe cADR cases.Objective: The aim of this study was to examine the immune reactions in cADR patients through the identification of the drug-specific proliferating cells by flow cytometric DLST (FCM-DLST).Methods: The peripheral blood mononuclear cells of 16 anticonvulsant-induced cADR patients were investigated by conventional DLST and a FCM-DLST protocol in which CFSE dilution and BrdU incorporation were combined. FCM-DLST allowed for the identification of the drug-specific proliferating cells in six cases. Three of these cases were DIHS cases, whereas there was one case of SJS, one case of maculopapular rash (MP), and one case of erythema multiforme (EM) among the six cases.Results: In FCM-DLST, drug-specific proliferating T cells were detected as CFSElow BrdUhigh cells. These cells corresponded to the cells incorporating 3H-thymidine in conventional DLST. Although CD4+ T-cell proliferation dominated the observed proliferation in most of the cases (in the recovery stage of the three DIHS cases, the MP case, and the EM case), drug-specific CD8+ cytotoxic T lymphocytes (CTLs) were detected, especially in the acute stages of the SJS case and one of the DIHS cases. There was a dramatic switch in the predominant drug-specific proliferating T-cell population in the course of one of the cases of DIHS in which CD8+ CTLs were predominant initially, whereas CD4+ T cells were predominant later. Moreover, drug-specific CD4+ CD25+ Foxp3+ regulatory T cells (Tregs) proliferated during the recovery stage in one DIHS case.Conclusions: FCM-DLST revealed that the cell proliferation detected by conventional DLST is a heterogeneous proliferation of both CD8+ CTLs and CD4+ T cells that likely includes Tregs. However, the number of cADR cases in this study was limited, which limits the conclusions that can be drawn from it.

A novel hairless mouse model for malignant melanoma - Corrected Proof

2011-12-15

Abstract: Background: An appropriate animal model for malignant melanoma could be a strong tool to develop biomarkers through analysis of melanomagenesis.Objective: Development of a novel animal model that spontaneously develops malignant melanoma with a high percentage.Methods: We crossed oncogenic RET (RFP–RET)-carrying transgenic mice of line 304/B6 (RET-mice) with hairless mice (hr/hr) and newly established hairless RFP–RET-transgenic mice of line 304-hr/hr (HL-RET-mice).Results: The HL-RET-mice developed hyperpigmented skin and benign melanocytic tumors without exception. More importantly, 63.8% (46/72) of the benign tumors were transformed to malignant melanoma in the HL-RET-mice. Mean time until the development of benign melanocytic tumors (2.4 months; n=102) in the HL-RET-mice was about half of that in the original RET-mice (4.6 months; n=20). Mean life span in the HL-RET-mice (9.7 months; n=38) was also significantly (p<0.01) shorter than that in the original RET-mice (10.8 months; n=20). Since early development of tumors could contribute to shortening of the research period, HL-RET-mice could be a useful model for analysis of melanomagenesis. We then found that the expression level of Mps one binder kinase activator-like-2B (Mobkl2b) in benign tumors was higher than that in malignant melanoma in HL-RET-mice. Expression level of MOBKL2B in malignant melanoma cell lines was also lower than that in non-malignant melanocytic cells in mice and humans, suggesting that MOBKL2B could be a novel marker for malignant melanoma.Conclusion: We established a novel hairless RET-transgenic mouse line spontaneously developing cutaneous malignant melanomas from benign melanocytic tumors. This mouse model may be useful to find new candidates of melanoma-related molecule.

WITHDRAWN: Analysis of interleukin-7 receptor-α gene single nucleotide polymorphism in Behcet's diseases - Corrected Proof

Koichiro Nakamura, Atsuko Neuchi, Tetsuya Tsuchida — 2011-03-18

This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause.The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy.

WITHDRAWN: Preface - Corrected Proof

Masutaka Furue — 2010-07-12

The publisher regrets that this article is an accidental duplication of an article that has already been published, doi:10.1016/j.jdermsci.2004.12.003. The duplicate article has therefore been withdrawn.

WITHDRAWN: Overview of Yusho - Corrected Proof

2010-07-12

The publisher regrets that this article is an accidental duplication of an article that has already been published, doi:10.1016/j.jdermsci.2004.12.004. The duplicate article has therefore been withdrawn.

WITHDRAWN: Behavior and toxic effects of PCBs and PCDFs in Yusho patients for 35 years - Corrected Proof

Yoshito Masuda, on behalf of the Study Group for Yusho — 2010-07-12

The publisher regrets that this article is an accidental duplication of an article that has already been published, doi:10.1016/j.jdermsci.2004.12.005. The duplicate article has therefore been withdrawn.

WITHDRAWN: Improvement in dioxin analysis of human blood and their concentrations in blood of Yusho patients - Corrected Proof

2010-07-12

The publisher regrets that this article is an accidental duplication of an article that has already been published, doi:10.1016/j.jdermsci.2004.12.006. The duplicate article has therefore been withdrawn.

WITHDRAWN: Blood chemistry, alpha-fetoprotein and hepatitis B surface antigen in Yusho - Corrected Proof

Hiroshi Tsuji, Yasuo Ito, on behalf of the Study Group for Yusho — 2010-07-12

The publisher regrets that this article is an accidental duplication of an article that has already been published, doi:10.1016/j.jdermsci.2004.12.007. The duplicate article has therefore been withdrawn.

WITHDRAWN: Cardiac, pulmonary and renal function in Yusho patients - Corrected Proof

2010-07-12

The publisher regrets that this article is an accidental duplication of an article that has already been published, doi:10.1016/j.jdermsci.2004.12.008. The duplicate article has therefore been withdrawn.

WITHDRAWN: Neurological signs and symptoms in patients with chronic PCB poisoning (Yusho accident) for more than 36 years - Corrected Proof

2010-07-12

The publisher regrets that this article is an accidental duplication of an article that has already been published, doi:10.1016/j.jdermsci.2004.12.009. The duplicate article has therefore been withdrawn.

WITHDRAWN: Complete blood cell counts and blood chemistry in Yusho - Corrected Proof

2010-07-12

The publisher regrets that this article is an accidental duplication of an article that has already been published, doi:10.1016/j.jdermsci.2004.12.010. The duplicate article has therefore been withdrawn.

WITHDRAWN: Ophthalmic findings in Yusho - Corrected Proof

2010-07-12

The publisher regrets that this article is an accidental duplication of an article that has already been published, doi:10.1016/j.jdermsci.2004.12.018. The duplicate article has therefore been withdrawn.

WITHDRAWN: Oral mucosa and dental findings in Yusho - Corrected Proof

2010-07-12

The publisher regrets that this article is an accidental duplication of an article that has already been published, doi:10.1016/j.jdermsci.2004.12.012. The duplicate article has therefore been withdrawn.

WITHDRAWN: Dermatological manifestations in Yusho: correlation between skin symptoms and blood levels of dioxins, such as polychlorinated dibenzofurans (PCDFs) and polychlorinated biphenyls (PCBs) - Corrected Proof

2010-07-12

The publisher regrets that this article is an accidental duplication of an article that has already been published, doi:10.1016/j.jdermsci.2004.12.016. The duplicate article has therefore been withdrawn.

WITHDRAWN: Sex ratio in the children of Yusho patients - Corrected Proof

2010-07-12

The publisher regrets that this article is an accidental duplication of an article that has already been published, doi:10.1016/j.jdermsci.2004.12.011. The duplicate article has therefore been withdrawn.

WITHDRAWN: Relationship of clinical symptoms and laboratory findings with blood levels of PCDFs in patients with Yusho - Corrected Proof

Yoshiyuki Kanagawa, Tomoaki Imamura, on behalf of the Study Group for Yusho — 2010-07-12

The publisher regrets that this article is an accidental duplication of an article that has already been published, doi:10.1016/j.jdermsci.2004.12.013. The duplicate article has therefore been withdrawn.

WITHDRAWN: The concepts of the new criteria for Yusho poisoning - Corrected Proof

2010-07-12

The publisher regrets that this article is an accidental duplication of an article that has already been published, doi:10.1016/j.jdermsci.2004.12.014. The duplicate article has therefore been withdrawn.

WITHDRAWN: Effects of dioxins on stress-responsive systems and their relevance to toxicity - Corrected Proof

2010-07-12

The publisher regrets that this article is an accidental duplication of an article that has already been published, doi:10.1016/j.jdermsci.2004.12.015. The duplicate article has therefore been withdrawn.

The current status and future direction of percutaneous peptide immunization against melanoma - Corrected Proof

Naohiro Seo, Masahiro Takigawa — 2010-07-12

Summary: Dendritic cell (DC)-based tumor immunotherapy is widely known to elicit protective anti-tumor immune responses, although the safety and effectiveness have yet to be thoroughly explored. We reported that a disruption in the stratum corneum barrier resulted in enhanced permeability and alterations in the skin immune system in such a way that epidermal Langerhans cells (LCs) functioned as vigorous antigen presenters for T helper (Th) cells and cytotoxic T lymphocytes (CTLs). In both human and murine models, topical application of melanoma-associated antigen peptides onto stratum corneum barrier-disrupted skin, specifically induced tumoricidal immune responses in vivo and in vitro accompanying an increased expression of MHC and co-stimulatory molecules on LCs. In addition, for reasons of simplicity, safety and effectiveness, percutaneous peptide application has demonstrated a certain degree of feasibility in clinical approach in patients with melanoma. In the future, resolution of some of the outstanding issues concerning the selection of the most effective adjuvants in combination with barrier disruption and depletion of regulatory T (Treg) cell-mediated immune suppression would appear as essential to improve percutaneous melanoma immunotherapy.

Significance of the expression of phosphorylated-STAT3, -Akt, and -ERK1/2 in several tumors of the epidermis - Corrected Proof

2010-07-12

In general, signal transduction pathways controlling cell division and proliferation are closely correlated. Signal transducer and activator of transcription-3 (STAT3), Akt, and extracellular signal-regulated kinase1/2 (ERK1/2) are main effector molecules that are associated with cell proliferation, apoptosis, differentiation, and malignant transduction . The aims of this study are to evaluate the expression of p-STAT3, p-Akt, and p-ERK1/2 in several tumors of epidermis, and evaluate correlations between their expressions and either the tumor size or metastasis of SCC.

Tricho-rhino-phalangeal type I syndrome and mental retardation: Identification of a novel mutation in the TRPS1 gene - Corrected Proof

Luz M. Gonzalez-Huerta, Sergio A. Cuevas-Covarrubias, Olga M. Messina-Baas — 2010-07-12

After Giedion reported the first case of tricho-rhino-phalangeal syndrome (TRPS) in 1966 , three types of TRPS (I, I, III) have been described in the literature . Pear-shape nose with bulbous tip, protruding ears, sparse hair, sparse eyebrows and long flat philtrum are common features to all TRPS types. Cone-shape epiphyses are the most characteristic radiographic findings, but they are not detected before 2 years of life. The 3 types of TRPS are due to affection of the TRPS1 gene, TRPS II is a contiguous gene syndrome that differs from TRPS I by the presence of mental retardation (MR) and multiple cartilaginous exostoses (MCE). MR ranges from borderline intelligence to profound MR . Microcephaly, large ears, micrognathia and lax skin frequently are present in TRPS II . TRPS III is similar to TRPS I except by more severe shortening of phalanges and metacarpals and short stature . TRPS I is inherited as an autosomal dominant trait and is due to molecular defects in the TRPS1 gene, TRPS II apparently follows autosomal dominant inheritance, but is due to a microdeletion in 8q24.1 region . Most TRPS II patients do not show a cytogenetically detectable deletion at 8q24.1; the majority of cases with TRPS II are sporadic . TRPS III is an allelic variant due to mutations in exon 6 of the TRPS1I gene . In the present study, we describe a TRPS I patient with MR and a novel 2bp deletion in the TRPS1 gene.

Targeting TNFα rapidly reduces density of dendritic cells and macrophages in psoriatic plaques with restoration of epidermal keratinocyte differentiation - Corrected Proof

Deborah J. Marble, Kenneth B. Gordon, Brian J. Nickoloff — 2010-07-12

Summary: Background: The cytokine network theory for psoriasis postulates a key role for TNFα in mediating inflammation and altered epidermal differentiation.Objective: This study defines responses following administration of adalimumab, a TNFα inhibitor, in pre-psoriatic skin (PN) and lesional psoriatic plaques (PP) skin.Methods: PN and PP skin before and after treatment were biopsied at days 2, 7, 28 and 84 (n=6 different patients). Cryosections were immunohistochemically stained to detect TNFα and other relevant markers in epidermal and dermal compartments. Detection of apoptosis utilized antibody specific for activated caspase 3. Semiquantitative assessments and statistical analysis was performed for each staining profile.Results: TNFα+ cells were increased in PP skin. PP skin was also characterized by a four-fold increase in number of CD68+ macrophages as well as eight-fold increase in CD11c+ dermal dendritic cells (DCs) compared to PN skin. By two-color immunofluorescence staining, both CD68+ cells as well as CD11c+ cells expressed TNFα. Following initiation of adalimumab therapy, CD11c+ cells, significantly decreased in PP skin at days 7, 28, and 84, while CD68+ and CD14+ cells decreased at days 28 and 84. Other markers for DCs (CD83, CD86) showed decreases at days 7, 28, and 84. Reduction in DCs, macrophages or T cells was not accompanied by increased activated caspase 3-positive cells. When a keratinocyte terminal differentiation marker was examined, adalimumab triggered rapid restoration of loricrin expression (beginning on day 2), with loss of aberrant differentiation marker, keratin 17 (K17).Conclusion: Adalimumab impacts dermal-based immunocytes, and the epidermal compartment also responds by restoration of normal differentiation without detectable apoptosis.

Rapid changes in substance P signaling and neutral endopeptidase induced by skin-scratching stimulation in mice - Corrected Proof

Junichi Yamaoka, Seiji Kawana — 2010-07-12

Summary: Background: Skin-scratching is a commonly seen behavior in patients with pruritus which sometimes exacerbates original lesions. Substance P (SP) signaling may play a predominant role in the pathophysiology induced by skin-scratching, however, it has not been well-elucidated.Objectives: To clarify changes in SP, its receptor NK-1R and a degradating enzyme neutral endopeptidase (NEP) induced by skin-scratching stimulation in mice.Methods: After skin-scratching stimulation was given to mice, changes in SP signaling were investigated as follows. Mast cell degranulation was examined with toluidine blue staining. SP-immunoreactive nerve fibers and the expressions of NK-1R and NEP were examined with immunofluorescence. Protein contents of SP and the enzymatic activity of NEP were examined with an ELISA and a colorimetric assay, respectively.Results: After skin-scratching stimulation, mast cells significantly degranulated within several minutes. SP-immunoreactive nerve fibers disappeared immediately from sensory nerve fibers, indicating the quick secretion and the depletion of SP. Both protein contents of SP and NEP activity in skin decreased dramatically soon after skin-scratching stimulation and thereafter they returned to the basal level within a week. The expression of NK-1R was significantly upregulated in epidermal basal keratinocytes after several days, in which NEP and NK-1R were well-coexpressed. Blocking NK-1R by an NK-1R antagonist suppressed scratching-induced decreases in SP-immunoreactive nerve fibers and in NEP activity.Conclusions: The present study clarified changing patterns of factors involved in SP signaling and NEP induced by skin-scratching stimulation. These findings provide basic and useful information to understand the pathophysiology of scratching-associated pruritic skin diseases.

Flavonolignans from Silybum marianum moderate UVA-induced oxidative damage to HaCaT keratinocytes - Corrected Proof

Alena Svobodová, Adéla Zdařilová, Daniela Walterová, Jitka Vostálová — 2010-07-12

Summary: Background: UV radiation from sunlight is a very potent environmental risk factor in the pathogenesis of skin cancer. Exposure to UV light, especially the UVA part, provokes the generation of reactive oxygen species (ROS), which induce oxidative stress in exposed cells. Topical application of antioxidants is a successful strategy for protecting the skin against UV-caused oxidative damage.Objective: In this study, silybin (SB) and 2,3-dehydrosilybin (DS) (1–50μmol/l), flavonolignan components of Silybum marianum, were tested for their ability to moderate UVA-induced damage.Methods: Human keratinocytes HaCaT were used as an appropriate experimental in vitro model, to monitor the effects of SB and DS on cell viability, proliferation, intracellular ATP and GSH level, ROS generation, membrane lipid peroxidation, caspase-3 activation and DNA damage.Results: Application of the flavonolignans (1–50μmol/l) led to an increase in cell viability of irradiated (20J/cm2) HaCaT keratinocytes. SB and DS also suppressed intracellular ATP and GSH depletion, ROS production and peroxidation of membrane lipids. UVA-induced caspases-3activity/activation was suppressed by treatment with SB and DS. Lower concentrations of both compounds (10μmol/l) significantly reduced cellular DNA single strand break formation.Conclusion: Taken together, the results suggest that these flavonolignans suppress UVA-caused oxidative stress and may be useful in the treatment of UVA-induced skin damage.

Nitric oxide levels in plasma and fibroblast cultures of psoriasis vulgaris patients - Corrected Proof

Anna Zalewska, Janina Wyczółkowska, Joanna Narbutt, Anna Sysa-Jędrzejowska — 2010-07-12

Nitric oxide (NO) is an active molecule generated in many cells including fibroblasts and endothelial cells, participating in psoriatic inflammatory processes. In context of dermal vascular dilatation and increased blood flow, being characteristic features of psoriasis, the contribution of NO deserves special attention, however literature data on NO production in psoriatic plaques are inconsistent .

Indications that topical l-carnitin-l-tartrate promotes human hair growth in vivo - Corrected Proof

Kerstin Foitzik, Edo Hoting, Ulrike Heinrich, Hagen Tronnier, Ralf Paus — 2010-07-12

l-Carnitine is essential for the intramitochondrial transport of long-chain fatty acids for β-oxidation . Recently, we have shown that l-carnitine-l-tartrate (CT) is able to increase hair shaft elongation and prolong anagen by upregulation of proliferation and downregulation of apoptosis in organ-cultured human scalp hair follicles . In order to test whether CT is also able to stimulate hair growth in human scalp hair follicles (HF) in vivo, we have performed a small, double-blind, randomised, placebo-controlled observational study of topical CT administration over 6 months twice a day. Penetration of CT into the skin, employing a liposomal hair tonic (Henkel, Düsseldorf, Germany) versus a conventional hair tonic was investigated, using the BUS (bovine udder model).

Identification of proteins involved in aggregation of human dermal papilla cells by proteomics - Corrected Proof

Xia Rushan, Hao Fei, Mou Zhirong, Wu Yu-zhang — 2010-07-12

Summary: Background: The dermal papilla is a major component of hair, which signals the follicular epithelial cells to prolong the hair growth process. To date, little is known about the significance of the specific protein(s) express in the dermal papilla cells (DPC) with regard to their aggregative behaviour.Objectives: To identify proteins involved in aggregative behaviour of DPC, we comparatively analyzed the proteome of cells with and without aggregative behaviour.Methods: A series of methods were used, including two-dimensional gel electrophoresis (2-DE), PDQuest software analysis of 2-DE gels, peptide mass fingerprinting based on matrix-assisted laser desorption/ionisation-time of flight-mass spectrometry (MALDI-TOF-MS), and NCBInr database searching, to separate and identify differentially expressed proteins. Western blotting and reverse transcriptase polymerase chain reaction (RT-PCR) were used to validate the differentially expressed proteins.Results: Image analysis revealed that averages of 618±22 and 568±47 protein spots were detected in passages 3 and 10 DPC, respectively. Twenty-four differential protein spots were measured with MALDI-TOF-MS. A total of 17 spots yielded good spectra, and 15 spots matched with known proteins after database searching. Western blotting confirmed that heat shocking protein 70 was up-regulated in passage 3 DPC. Over-expression of mitochondrial ribosomal protein S7 was confirmed by RT-PCR, indicating that they are involved in aggregation of DPC through some signaling pathway.Conclusions: The clues provided by the comparative proteome strategy utilized here will shed light on molecular mechanisms of DPC in aggregative behaviour.

Associations of IL-2 and IL-4 gene polymorphisms with psoriasis in the Korean population - Corrected Proof

2010-07-12

Summary: Background: Psoriasis is association with an overexpression of T-helper cell type 1(Th1) cytokines and relative underexpression of Th2 cytokines. The cytokine production is under genetic control, and certain allelic variants of cytokine genes are associated with higher or lower cytokine production in vitro and in vivo.Objectives: We aimed to evaluate association of cytokine genes polymorphisms with psoriasis in the Korean population.Methods: We investigated the polymorphisms of IL-2 −330, IL-4 −590, IL-4 receptor +1902, IL-10 −1082 and −819, and IFN-γ intron 1 in 114 psoriasis patients and 281 healthy normal controls in Korean.Results: IL-2 −330*G and IL-4 −590*C alleles significantly increased in psoriasis patients, especially late-onset group, compared to the control. The combined effect of IL-2 −330*G and IL-4 −590*C showed that the positive combination of IL-2 −330*G and IL-4 −590*C alleles were more significantly associated with the late-onset group of psoriasis patients than the controls.Conclusions: These results suggest that the genetic polymorphisms of IL-2 and IL-4 genes can be susceptible to psoriasis in Korean, especially late-onset psoriasis group.

Topical application of laminin-332 to diabetic mouse wounds - Corrected Proof

2010-07-12

Summary: Background: Keratinocyte migration is essential for wound healing and diabetic wound keratinocytes migrate poorly. Keratinocyte migration and anchorage appears to be mediated by laminin-332 (LM-332). Impaired diabetic wound healing may be due to defective LM-332 mediated keratinocyte migration.Objective: To evaluate LM-332 expression in diabetic (db/db) and control (db/−) mice and to test LM-332 wound healing effects when applied to mouse wounds.Methods: LM-332 expression in mouse wounds was evaluated using immunohistochemistry. LM-332 wound healing effects were evaluated by directly applying soluble LM-332, a LM-332 biomaterial, or a control to mouse wounds. Percent wound closure and histology score, based on healing extent, were measured.Results: Precursor LM-332 expression was markedly reduced in db/db when compared to db/− mice. In vitro, soluble LM-332 and LM-332 biomaterial demonstrated significant keratinocyte adhesion. In vivo, soluble LM-332 treated wounds had the highest histology score, but significant differences were not found between wound treatments (p>0.05). No differences in percentage wound closure between treatment and control wounds were found (p>0.05).Conclusion: The db/db wounds express less precursor LM-332 when compared to db/−. However, LM-332 application did not improve db/db wound healing. LM-332 purified from keratinocytes was primarily physiologically cleaved LM-332 and may not regulate keratinocyte migration. Application of precursor LM-332 rather than cleaved LM-332 may be necessary to improve wound healing, but this isoform is not currently available in quantities sufficient for testing.

A spectrum of mutations in keratins K6a, K16 and K17 causing pachyonychia congenita - Corrected Proof

2010-07-12

Summary: Background: Pachyonychia congenita (PC) is a rare autosomal dominant keratin disorder, subdivided into two major variants, PC-1 and PC-2. Predominant characteristics include hypertrophic nail dystrophy, focal palmoplantar keratoderma and oral leukokeratosis. Multiple steatocystomas that develop during puberty are a useful feature distinguishing PC-2 from PC-1. At the molecular level it has been shown that mutations in keratin K6a or K16 cause PC-1 whereas those in K6b or K17 lead to PC-2.Objective: To identify mutations in 22 families presenting with clinical symptoms of either PC-1/focal non-epidermolytic palmoplantar keratoderma (FNEPPK) or PC-2.Methods: Mutation analysis was performed on genomic DNA from PC patients by direct sequencing.Results: Here, we report four new missense and five known mutations in K6a; one new deletion and three previously identified missense mutations in K16; plus one known mutation in K17.Conclusion: With one exception, all these heterozygous mutations are within the highly conserved helix boundary motif regions at either end of the keratin rod domain. In one sporadic case, a unique mutation in K16 resulting in deletion of 24bp was found within the central rod domain, in a child with a phenotype predominantly consisting of focal plantar keratoderma. The identification of mutations in cases of PC is prerequisite for future development of gene-specific and/or mutation-specific therapies.