Autoantibodies to nuclear valosin-containing protein-like protein: systemic sclerosis-specific antibodies revealed by in vitro human proteome.

OBJECTIVES: To identify and characterize undescribed systemic sclerosis (SSc)-specific autoantibodies targeting nucleolar antigens and to assess their clinical significance. METHODS: We conducted proteome-wide autoantibody screening (PWAS) against serum samples from SSc patients with nucleolar patterned anti-nuclear antibodies (NUC-ANAs) of specific antibodies (Abs) unknown, utilizing wet protein arrays fabricated from in vitro human proteome. Controls included SSc patients with already-known SSc-specific autoantibodies, patients with other connective tissue diseases, and healthy subjects. The selection of nucleolar antigens was performed by database search in the Human Protein Atlas. The Presence of autoantibodies was certified by immunoblots and immunoprecipitations. Indirect immunofluorescence assays on HEp-2 cells were also conducted. Clinical assessment was conducted by retrospective review of electric medical records. RESULTS: PWAS identified three candidate autoantibodies, including anti-nuclear valosin-containing protein-like (NVL) Ab. Additional measurements in disease controls revealed that only anti-NVL Abs are exclusively detected in SSc. Detection of anti-NVL Abs was reproduced by conventional assays such as immunoblotting and immunoprecipitation. Indirect immunofluorescence assays demonstrated homogeneous nucleolar patterns. Anti-NVL Ab-positive cases were characterized by significantly low prevalence of diffuse skin sclerosis and interstitial lung disease, compared with SSc cases with NUC-ANAs other than anti-NVL Abs, such as anti-U3-RNP and anti-Th/To Abs. CONCLUSION: Anti-NVL Ab is an SSc-specific autoantibody associated with a unique combination of clinical features, including limited skin sclerosis and lack of lung involvement.

Physiological effects of appropriate washing on xerotic skin.

BACKGROUND: Daily skin care is important for treatment of skin diseases, but few studies have reported on appropriate washing methods. AIM: This study aimed to provide guidance on washing techniques and examine changes in skin condition after using the recommended washing technique and foaming-type skin cleanser in patients with atopic or asteatotic dermatitis. METHODS: An internet-based questionnaire survey on skin symptoms and cleaning methods was conducted. Further, a left-right comparative, nonrandomized trial was performed in 19 patients with asteatotic or atopic dermatitis and xerosis. Participants were instructed to wash with a cotton towel and their normal cleanser during Weeks 1-4 and with bare hands or a cotton towel and the recommended foaming-type cleanser during Weeks 5-8. RESULTS: The survey revealed that the degree of lathering differed depending on the cleaning tool. In the trial, scores for erythema, desquamation, and xerosis in the lower legs were significantly reduced after 4 weeks compared with scores at the start. Between Weeks 4 and 8, scores for erythema, xerosis, and pruritus in the inner forearm on the side washed with bare hands and scores for xerosis, pruritus, and excoriation on the side washed with a cotton towel were significantly reduced. A significant increase was noted in stratum corneum ceramide content on both left and right inner forearms, whereas a significant decrease was noted in stratum corneum thymus and activation-regulated chemokine level ratios in the lower legs on both sides. CONCLUSIONS: Xerotic skin disease symptoms can be improved using appropriate body washing methods.

Bioactive substances in the stratum corneum of the epidermis found as indicators of skin damage due to sun exposure.

BACKGROUND/PURPOSE: Although an inflammatory response upon acute injury caused by ultraviolet radiation (UV) can be observed immediately, the influence of long-term, repetitive low-dose UV exposure on the skin cannot be precisely perceived, making early detection of chronic damage difficult. This study investigated bioactive substances in the stratum corneum as a potential early and sensitive indicator of the influence of sun exposure on the skin using receiver operating characteristic (ROC) analysis. METHODS: Receiver operating characteristic analysis was performed to assess the responsiveness of cytokines [interleukin (IL)-1α, IL-1 receptor antagonist (IL-1ra), IL-10, tumor necrosis factor (TNF)-α], BCL2-associated protein X (Bax), Toll-like receptor (TLR)3, and TLR4 in the stratum corneum of healthy people exposed (dorsum of the hand) and unexposed (inner arm) to UV. Sunscreen was applied to patients with photodermatosis for 4 weeks to evaluate changes in IL-1ra/IL-1α, TNF-α, Bax, and TLR3 levels after sunscreen application, as these molecules exhibited high responsiveness to sun exposure according to ROC analysis. In addition, IL-1ra, IL-1α, and IL-10 levels were quantified by enzyme-linked immunosorbent assay, and TNF-α, Bax, TLR3, and TLR4 levels were semi-quantitatively assessed by immunocytochemistry. RESULTS: Receiver operating characteristic analysis identified IL-1ra/IL-1α, TNF-α, Bax, and TLR3 in the stratum corneum as highly responsive to sun exposure. Moreover, in participants, including patients with photodermatosis, IL-1ra/IL-1α, TNF-α, and Bax levels decreased significantly after sunscreen application. CONCLUSION: The results revealed that IL-1ra/IL-1α, TNF-α, and Bax in the stratum corneum represent sensitive indicators of the influence of sun exposure on the skin.

Optimal application method of a moisturizer on the basis of skin physiological functions.

BACKGROUND: Clinical studies have clarified the usefulness of moisturizers for dry skin diseases. However, few reports exist on the appropriate application of moisturizers with respect to the skin physiological functions. AIMS: To clarify the optimal moisturizer application method on the basis of skin physiological functions. METHODS: This study investigated the appropriate time, dose, and frequency of moisturizer application from the perspective of skin physiology. In healthy subjects, the stratum corneum water content (SCW) was compared between different moisturizer application times (immediately [≤5 min] and 90 min after bathing), doses (0.5, 1.0, and 2.0 mg/cm2 ), and frequencies (once and twice daily). Thereafter, patients with dry skin were treated with the moisturizer once or twice daily for 8 weeks at the time, and application dose was determined to be optimal for the healthy subjects; the moisturizing effect was evaluated based on the SCW, trypsin activity, and desmoglein 1 localization score in the stratum corneum. RESULTS: In healthy subjects, compared to at control sites, the SCW was significantly higher at sites treated with the moisturizer immediately after bathing, with 1.0 and 2.0 mg/cm2 of the moisturizer, and with once- and twice-daily applications. In patients with dry skin, the SCW was significantly higher compared to control sites and the desmoglein 1 localization score was significantly lower after 8 weeks only when the moisturizer was applied twice daily. CONCLUSIONS: Moisturizer application of ≥1.0 mg/cm2 twice daily (immediately after bathing at night and in the morning) had a moisturizing effect, as verified from the skin physiological functions.

Effect of Vitamin-Containing Amino Acid Supplements on Menopausal Symptoms and Age-Related Skin Changes: A Randomised, Double-Blind, Placebo-Controlled Study.

INTRODUCTION: Women aged ≥ 45 years are known to experience various menopausal symptoms due to reduced oestrogen levels. Changes associated with cutaneous ageing, such as wrinkles and sagging, are also prominent in women in this age group. Numerous studies have evaluated the usefulness of individual basic nutrients and essential components, such as amino acids and vitamins, in menopausal women. The aim of this study was to comprehensively assess the effects of supplements containing amino acid and vitamin combinations on menopausal symptoms and age-related changes in the skin. METHODS: This is a randomised, placebo-controlled study on the effects of a supplement containing three types of amino acids [leucine, glutamine and arginine (LGA)] and 11 types of vitamins in 37 middle-aged women. The participants ingested either the test substance or a placebo twice daily for 8 consecutive weeks, at the end of which time the efficacy and safety of the test substance were assessed based on subjective symptoms of fatigue, Simplified Menopausal Index (SMI) score, the grade and number of wrinkles at the corners of the eyes, results of an analysis of the stratum corneum of the cheek and blood test results. RESULTS: None of the participants experienced adverse events, and all items assessed in our survey of subjective symptoms of fatigue showed a significant decrease in the test substance group (LGA group). Additionally, the SMI score of the LGA group was significantly lower than that of the placebo group. The LGA group showed significant improvement in the grade of the wrinkles in the corners of the eyes and the number of wrinkles, and the stratum corneum cell area declined significantly in the LGA group compared to the placebo group at the end of the study. CONCLUSION: These results indicate that the vitamin-containing amino acid supplement improved menopausal symptoms and age-related changes in the skin (wrinkles). TRIAL REGISTRATION: UMIN000029830.

Filaggrin-gene mutation has minimal effect on the disease severity in the lesions of atopic dermatitis.

Loss-of-function mutations of filaggrin (FLG) gene (FLG) are the strongest known genetic risk factor for atopic dermatitis (AD). It is still debatable how FLG gene mutations and the resulting abnormal amount of FLG protein contribute to skin barrier function and symptoms of AD. In this study, we examined the effects of loss-of-function mutations of FLG gene on the severity of skin lesions and skin barrier function in 55 patients with AD by evaluating eight patients with AD with FLG gene mutations and 47 patients with AD without mutations. The results showed that the FLG gene mutation did not affect the duration of AD, severity of AD, degree of local inflammatory symptoms, skin water content and trans-epidermal water loss of the lesions. Next, in these eight mutation carriers and the 47 non-carriers, stratum corneum was collected from the three site of skin lesions using tape-stripping method, and the amounts of FLG protein and total amino acid contained in the stratum corneum was measured to investigate the effect of the FLG gene mutation on the amount of FLG gene product in the local lesion. FLG abnormalities had little effect on FLG protein and total amino acid content in the stratum corneum in the lesional skin. The amount of the FLG products, especially amino acids derived from FLG, in the stratum corneum of AD lesional skin is influenced by development of dermatitis. The results obtained from this study supports that the activation of Th2-dominant inflammatory cells, together with FLG abnormality, plays a role in suppressing the production of FLG in skin lesions.

Novel petrolatum-based ointment that is highly moisturizing and has superior usability with increased adherence in patients with facial dry skin.

BACKGROUND: Petrolatum is often used to suppress water evaporation from the skin surface. However, its stickiness, shininess, and other factors make it inconvenient for continuous use. OBJECTIVE: To compare the effectiveness and usability between a newly developed petrolatum-based ointment (NOV® II Moisture Balm) and white petrolatum. METHODS: Twenty-nine subjects with atopic dermatitis or xeroderma with symptoms of dry skin applied NOV® II Moisture Balm on the right sides of the face and white petrolatum on the left side, respectively, for 8-12 weeks. The skin conditions (erythema, exudate/crusting, dryness, and itch) were scored, stratum corneum water content and transepidermal water loss (TEWL) were measured, and the free amino acid and thymic stromal lymphopoietin (TSLP) levels in the stratum corneum were analyzed before and end of the trial. RESULTS: Dryness, exudate/crusting, and TEWL decreased significantly on both the white petrolatum and the NOV® II Moisture Balm sides, while stratum corneum water content and the free amino acid levels in the stratum corneum increased significantly. On the NOV® II Moisture Balm side, erythema and the TSLP levels decreased significantly. In a questionnaire on usability, the subjects felt NOV® II Moisture Balm spread better and were less sticky and shiny than white petrolatum, and more subjects wanted to continue using NOV® II Moisture Balm. CONCLUSIONS: NOV® II Moisture Balm was equivalent to white petrolatum in moisturizing and improving the physiologic functions of the skin, but had superior usability.

Physiological and microbiological verification of the benefit of hair washing in patients with skin conditions of the scalp.

BACKGROUND: It is unclear whether hair washing is effective against scalp eruption and pruritus caused by seborrheic dermatitis or psoriasis vulgaris. AIMS: To assess whether a proper hair-washing regimen, including the use of antibacterial shampoo, can ameliorate scalp eruption symptoms and alter the composition of the scalp microflora. METHODS: Eighteen patients with seborrheic dermatitis or psoriasis vulgaris scalp eruptions were instructed in proper techniques of daily hair washing, rinsing, and shampooing, which they underwent for 12 weeks. They used control shampoo in weeks 1-4 and 9-12, and an antibacterial shampoo during weeks 5-8. At the start of the test period and at weeks 4, 8, and 12, we assessed scalp symptoms (erythema, scaling/desquamation, dryness, itchiness, and scratching scars); microbial DNA levels from lesion and nonlesion areas; and levels of interleukin (IL)-1α, IL-1ra, and total protein in the scalp's horny layer. RESULTS: Compared to baseline values, scaling/desquamation and itchiness improved significantly at weeks 8 and 12. Other observed skin symptoms also improved over time. Malassezia colonization levels in lesion and nonlesion areas decreased gradually; the decrease was significant at week 8 in lesion areas and at weeks 4 and 8 in nonlesion areas. Bacterial colonization levels also decreased gradually, achieving significance in lesion areas at week 4. Gradual decreases in IL-1ra/IL-1α level showed statistical significance at weeks 4 and 12, while the protein quantity significantly decreased at week 12. CONCLUSION: Proper hair washing improved scalp condition symptoms, and possibly the underlying etiology.

Clinical studies of the treatment of facial atrophic acne scars and acne with a bipolar fractional radiofrequency system.

Few clinical studies have examined the utility of bipolar fractional radiofrequency (FRF) therapy as a treatment for atrophic acne scars and active acne in people with darker skin. This study was designed to compare the safety and efficacy of bipolar FRF therapy as a treatment for atrophic acne scars and acne vulgaris. Twenty-three Japanese patients with atrophic acne scars and mild to severe acne on both cheeks were treated with a bipolar FRF system (eMatrix; Syneron, Yokneam Illit, Israel). Five treatment sessions were carried out at 1-month intervals, and the patients were followed up for 3 months after the final treatment. Assessments of scar severity and the number of acne lesions and 3-D in vivo imaging analysis were performed. Evaluations of the treatment outcomes and their effects on the patients' quality of life (QOL) were also carried out. We demonstrated that the improvement in scar volume was marked in the patients with mild scars and was at least moderate in 23 (57.5%) of the treated areas. With regard to the number of acne lesions, the treated areas exhibited significantly fewer lesions compared with the baseline at each time point (P < 0.05). The patients' assessments of the treatment outcomes and their QOL indicated that both had improved significantly by the end of the study. Furthermore, significant reductions in the patients' sebum levels, skin roughness and scar depth were observed. Bipolar FRF treatment significantly improved the atrophic acne scars and acne of Japanese patients and had minimal side-effects.

Clinical evaluation of glycolic acid chemical peeling in patients with acne vulgaris: a randomized, double-blind, placebo-controlled, split-face comparative study.

BACKGROUND: Glycolic acid (GA) peels are frequently performed as adjuvants to the treatment of facial acne. There have been few clinical trials reported of GA peels for acne in people with darker skin. OBJECTIVES: To determine the safety and efficacy of GA peels in the treatment of moderate acne vulgaris in Asian skin. METHODS: In this prospective, randomized, double-blind, placebo-controlled, split-face clinical trial, 26 patients with moderate acne were treated with 40% GA (pH 2.0) on half of the face and placebo on the other half. The procedure was performed five times at 2-week intervals. RESULTS: The GA sides had statistically significant reductions in acne lesions at each time point from baseline values. There were statistically significant differences between the GA and placebo sides. The GA sides had better responses for noninflammatory lesions than for inflammatory lesions. In bioengineering measurements, sebum levels were statistically significantly reduced after the initiation of therapy on both sides at weeks 8 and 10, but there were no statistically significant differences between the two sides. CONCLUSION: Forty percent GA peels significantly improved moderate acne in this study. It is effective and safe in Asians.

Chemokine receptor CXCR4 is a novel marker for the progression of cutaneous malignant melanomas.

The CXCR4/CXCL12 pathway has recently been reported to be involved in stimulating the metastasis of many different neoplasms, in which CXCR4 activates various phenomena such as chemotaxis, invasion, angiogenesis and proliferation. The purpose of this study was to analyze a possible association between the expression of chemokine receptors CXCR4, CCR6 and CCR7 with the clinicopathological features of cutaneous malignant melanoma, and to assess the usefulness of these chemokine receptors for diagnosis and prognosis. In our study, a percentage of immunoexpression of both CXCR4 and its ligands CXCL12 was associated with high clinical risk. In contrast, the patients with a low immunoexpression of CXCR4 and CXCL12 had low clinical risk. CCR6 and CCR7 immunoexpressions were also correlated with some clinical parameters, but seemed no more useful than CXCR4. These data suggest that the assessment of CXCR4 immunoexpression is a novel tool for predicting tumor aggressiveness in malignant melanomas, and in particular, a high immunoexpression percentage of CXCR4 and CXCL12 might be a sign of a poor prognosis.

Effect of sequential application of topical adapalene and clindamycin phosphate in the treatment of Japanese patients with acne vulgaris.

The efficacy of combined therapy with a retinoid and antibiotic for Japanese patients with acne vulgaris remains to be established. Further, maintenance strategies limiting the use of topical retinoids must be identified. The objectives of this study are to determine the efficacy of sequential application of topical adapalene and clindamycin phosphate and to assess the impact of this regimen on patients' quality of life. Sixty-six patients were recruited. The regimen comprised two phases. For the 4-week initial treatment, 1% clindamycin phosphate gel was applied twice daily and 0.1% adapalene gel, once. In the 4-week maintenance phase, patients were randomly assigned to the OD group (adapalene applied once daily) or the TW group (adapalene applied once daily on 2 days per week). The acne severity score, lesion counts, microcomedone count, and sebum amount were measured. Quality of life (QOL) was assessed using Skindex-16. All parameters improved significantly by week 4 of initial treatment. No statistically significant differences were found in the improvement of clinical findings between the groups. All QOL scores improved significantly and did not significantly differ between the groups. Our regimen may enable clinical control of acne in Japanese patients and improve their QOL. For limiting retinoid use, weekly application of adapalene during maintenance is suitable.

Combination effects of cosmetic moisturisers in the topical treatment of acne vulgaris.

Cosmetic moisturisers were applied to one side of the face of 18 male Japanese patients with acne vulgaris who were treated with a topical administration of adapalene and clindamycin phosphate gels. We assessed the alleviating effect of the moisturisers on the side effects of the treatment. The severity of acne and the number of inflammatory and non-inflammatory lesions were measured at 0, 2, and 4 weeks. The water content in the stratum corneum and transepidermal water loss were measured by comparing the moisturiser-treated and untreated sides of the face. The sensation of skin dryness and irritation on both sides of the face were assessed by the subjects. We observed that the use of moisturisers did not impact the efficacy of the standard topical treatment and they significantly improved the water content in the stratum corneum and the sensation of dryness. These results suggested that the use of moisturisers in combination with the standard topical treatment may improve adherence to therapy by alleviating the sensation of dryness.

Effect of chemical peeling on photocarcinogenesis.

Chemical peeling is one of the dermatological treatments available for certain cutaneous diseases and conditions or improvement of cosmetic appearance of photo-aged skin. We assessed the photo-chemopreventive effect of several clinically used chemical peeling agents on the ultraviolet-irradiated skin of hairless mice. Chemical peeling was done using 35% glycolic acid dissolved in distilled water, 30% salicylic acid in ethanol, and 10% or 35% trichloroacetic acid in distilled water at the right back of ultraviolet-irradiated hairless mice every 2 weeks for glycolic acid, salicylic acid and 10% trichloroacetic acid, and every 4 weeks for 35% trichloroacetic acid for a total of 18 weeks after the establishment of photo-aged mice by irradiation with ultraviolet B range light three times a week for 14 weeks at a total dose of 6.66 J/cm(2) . Tumor formation was assessed every week. Skin specimens were taken from treated and non-treated area for evaluation under microscopy, evaluation of p53 expression and mRNA expression of cyclooxygenase-2. Serum level of prostaglandin E(2) was also evaluated. All types of chemical peeling reduced tumor formation in treated mice, mostly in the treated area but also in the non-treated area. Peeling suppressed retention of p53-positive abnormal cells and reduced mRNA expression of cyclooxygenase-2 in treated skin. Further, serum prostaglandin E(2) level was decreased in chemical peeling treated mice. These results indicate that chemical peeling with glycolic acid, salicylic acid and trichloroacetic acid could serve tumor prevention by removing photo-damaged cells.

Preventive effect of chemical peeling on ultraviolet induced skin tumor formation.

BACKGROUND: Chemical peeling is one of the dermatological treatments available for certain cutaneous diseases and conditions or improvement of cosmetic appearance of photoaged skin. OBJECTIVES: We assessed the photochemopreventive effect of several clinically used chemical peeling agents on the ultraviolet (UV)-irradiated skin of hairless mice. METHODS: Chemical peeling was done using 35% glycolic acid dissolved in distilled water, 30% salicylic acid in ethanol, 10% or 35% trichloroacetic acid (TCA) in distilled water at the right back of UV-irradiated hairless mice every 2 weeks in case of glycolic acid, salicylic acid, and 10% TCA and every 4 weeks in case of 35% TCA for totally 18 weeks after the establishment of photoaged mice by irradiation with UVA+B range light three times a week for 10 weeks at a total dose of 420 J/cm(2) at UVA and 9.6 J/cm(2) at UVB. Tumor formation was assessed every week. Skin specimens were taken from treated and non-treated area for evaluation under microscopy, evaluation of P53 expression, and mRNA expression of cyclooxygenase (COX)-2. Serum level of prostaglandin E(2) was also evaluated. RESULTS: All types of chemical peeling reduced tumor formation in treated mice, mostly in the treated area but also non-treated area. Peeling suppressed clonal retention of p53 positive abnormal cells and reduced mRNA expression of COX-2 in treated skin. Further, serum prostaglandin E(2) level was decreased in chemical peeling treated mice. CONCLUSIONS: These results indicate that chemical peeling with glycolic acid, salicylic acid, and TCA could serve tumor prevention by removing photodamaged cells.