Superficial intense focused ultrasound on periorbital wrinkle.

The periorbital wrinkles are easily perceived evidence of aging, so become a major concern for many patients. Various treatments have been attempted to improve periorbital wrinkles, but the need for new treatments that are less invasive and more effective is still high. In this study, we evaluated the safety, clinical and histological effects of intense focused ultrasound using only a 1.5 mm transducer in the management of periorbital wrinkles. Ten adult Korean females were enrolled. The treatment effect and safety profile were evaluated up to 3 months after 1 session of IFUS treatment on the periorbital wrinkles. The mean subjective satisfaction score was 3.2 ± 0.79 (mean ± standard deviation) by 5- point scale. The mean objective clinical improvement score was highest in the fine wrinkle on the crow's feet area and lowest in the deep wrinkles of the infraorbital and crow's feet area. Histometrically, increase of collagen and elastic fiber density was observed in the all layers of dermis. No serious side effects occurred after the treatment. In conclusion, intense focused ultrasound treatment using a 1.5-mm transducer alone can significantly improve periocular wrinkles after a single treatment with a good safety profile.

Dermal Remodeling of Burn Scar by Fractional CO2 Laser.

BACKGROUND: Ablative CO2 fractional lasers have recently been introduced for burn scar treatment because of pronounced clinical outcomes with fewer treatment sessions than nonablative fractional laser. OBJECTIVE: This study was conducted to observe clinical as well as histologic changes of burn scars after treatment with CO2 fractional laser. MATERIALS AND METHODS: Eleven patients (one female and 10 males, aged 31-59 years) with skin phototypes III to V with burn scars received 10 sessions of fractional CO2 laser treatments (UltraPulse(®) Encore; Lumenis, Santa Clara, CA, USA) over an average 5-week interval. Two passes were performed using the following parameters: deep FX mode, 12.5-30 mJ, with a density setting of 5-10 %. Clinical evaluations by three blinded dermatologists were obtained at baseline and at 6 months after the final treatment via photographs using the Vancouver scar scale (VSS). Skin biopsies were done on four patients before and after treatment. RESULTS: All patients showed clinical improvement in their scars with significant decrease in VSS. Histologic findings showed the changes in the upper dermis with newly formed dermal papilla. This characteristic upper dermis change was presented as improvement in surface smoothness and skin tension clinically. Postinflammatory hyperpigmentation and itching sensation were the most common adverse effects. CONCLUSION: Burn scar treatment by fractional CO2 laser is effective by forming new collagen fibers mainly in the upper dermis. LEVEL OF EVIDENCE IV: 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 www.springer.com/00266 .

Treatment of dilated pores with 1410-nm fractional erbium-doped fiber laser.

Dilated pores can be an early sign of skin aging and are a significant cosmetic concern. The 1410-nm wavelength is optimal for superficial dermal treatments up to 650 µm deep. The aim of the present study was to evaluate the clinical effectiveness and safety of the fractional erbium-doped fiber 1410-nm laser in the treatment of dilated pores. Fifteen patients with dilated facial pores underwent three laser treatments at 3-week intervals. Posttreatment skin responses and side effects were assessed at treatment and follow-up visits by study physicians. Clinical effectiveness of treatment was assessed by both study physicians and patients 3 months after the final laser treatment using a quartile grading scale. Histological examination was performed using biopsy samples taken at baseline (pretreatment) and 3 months after the last treatment. This study showed that greater than 51 % improvement in dilated pores was demonstrated in 14 of 15 patients after three sessions of laser treatments. Improvements in skin texture, tone, and smoothness were reported in all patients. Treatment was well tolerated in all patients, with no unanticipated side effects. This study demonstrates that the 1410-nm fractional erbium fiber laser is effective and safe for treatment of dilated facial pores in Fitzpatrick skin types III-IV.

Intense focused ultrasound for facial tightening: histologic changes in 11 Patients.

INTRODUCTION: Intense focused ultrasound (IFUS) is a novel modality for treating skin laxity that produces thermal effects at various depths while sparing the overlying tissue. This study assessed histologic changes and the safety and efficacy of intense focused ultrasound (Doublo(TM), HIRONIC Co., Sungnam, Korea) for tightening of facial skin in Asian patients. METHODS: Eleven patients with facial laxity were treated with IFUS and evaluated before and after treatment. Mean age was 46 years (range, 35-64 years). Two available hand-pieces with different focal depths (3 mm and 4.5 mm) were used with three to five passes 1-2 mm apart. Outcome assessment included photographic evaluation by two blinded investigators, skin biopsies before and two months after treatment, and patient satisfaction. RESULTS: Subjective and objective analyses showed 63.6% and 72.7% improvement at the two-month evaluation, respectively. Histologic evaluation by hematoxylin and eosin (H&E) and Masson's trichrome staining showed increased collagen fibers in the lower dermis and between fat layers. DISCUSSION AND CONCLUSIONS: Intense focused ultrasound can be used as a non-invasive skin tightening technique in Asian patients. It induced collagen generation in the dermis and fat layers and was effective and safe in our study population.

The efficacy of a microwave device for treating axillary hyperhidrosis and osmidrosis in Asians: a preliminary study.

A microwave-based device has been developed to treat axillary hyperhidrosis by selectively heating the interface between the skin and underlying fat in the axilla. This study was conducted to evaluate the efficacy and safety of microwave-based devices for axillary hyperhidrosis and osmidrosis in Asians. Eleven patients (8 females and 3 males, age range 20-52 years, mean age 37.6 years) with axillary hyperhidrosis or osmidrosis were enrolled, treated with the microwave-based device, and followed up for 7 months. Procedure efficacy, patient satisfaction, and treatment safety were assessed. The clinical records were reviewed and the patients were interviewed individually at follow-up visits or via telephone. Evaluation of sweating showed at least a 2-point drop or greater in hyperhidrosis disease severity scale (HDSS) in 83.3% subjects (10/12 axillae) as measured at the 7-month follow-up. Of 16 axillae with osmidrosis, 93.8% (15/16 axillae) showed good to excellent results. Histologic findings also showed destruction of eccrine and apocrine glands that were replaced with fibrosis. Regarding safety, altered sensation of arms developed in one case that resolved after 3 months. This novel microwave-based treatment appears to be effective and well tolerated for the treatment of axillary hyperhidrosis and osmidrosis in Asians.

Extramammary paget disease: minimal surgical therapy.

BACKGROUND: Extramammary Paget disease (EMPD) is an uncommon malignant neoplasm affecting apocrine gland-bearing skin which usually occurs in the anogenital area of patients older than 50 years. Although Mohs micrographic surgery (MMS) is recommended for the treatment of EMPD, wide local excision has also been performed by many other surgeons including dermatosurgeons. However, the extent of an adequate resection margin is still under debate. OBJECTIVE: The efficacy of minimal surgical therapy consisting of a wide excision combined with preoperative multiple scouting biopsies and postoperative topical imiquimod was investigated for the treatment of EMPD in Korean patients. METHODS: Between 2006 and 2012, 10 patients with primary EMPD were treated with wide surgical excision, with a surgical margin of less than 2.5 cm. Multiple preoperative scouting biopsies and postoperative topical imiquimod were also performed to delineate the lesional boundaries and to reduce the recurrence rate. RESULTS: During the 6-year follow-up period, complications and recurrences were not observed. CONCLUSION: Minimal surgical therapy may be an effective alternative when MMS is unavailable.

A intense-focused ultrasound tightening for the treatment of infraorbital laxity.

BACKGROUND: Infraorbital laxity is a common problem that increases with age. Blepharoplasty with lipectomy is a very commonly performed surgical procedure to treat this problem; however, it is invasive and is associated with the potential for re-emergence. Therefore, young patients may prefer a non-surgical procedure rather than to a surgical procedure. Intense-focused ultrasound (IFUS) has emerged as an effective, non-surgical, tissue-tightening procedure. OBJECTIVE: This study assessed the safety and efficacy of IFUS (Ulthera system, Ulthera Inc, Mesa, AZ, U.S.A.) for facial tightening in Asian patients with infraorbital laxity. METHODS: We studied 15 patients who were treated with an IFUS device applied to both lower eyelids. The primary outcome measure was an objective improvement in a paired comparison of pre-treatment and post treatment (6 months) photographs. A secondary outcome measure was patient satisfaction as measured by a questionnaire. RESULTS: The mean patient age was 50 years (range, 27-69). All patients received one to two treatments with intense-focused ultrasound. All patients in the study experienced both subjective and objective improvement. CONCLUSION: IFUS can be used as a non-invasive, skin-tightening procedure for infraorbital laxity. No serious, permanent, or delayed side effects were noted up to 6 months post treatment. Thus, this procedure can be effective and safe in the treatment of decreased laxity of the lower eyelids.

Treatment of striae distensae combined enhanced penetration platelet-rich plasma and ultrasound after plasma fractional radiofrequency.

BACKGROUND: Striae distensae (striae alba) is a challenging cosmetic problem for which various treatment modalities have been applied. However, the treatment of striae distensae has not been satisfactory. OBJECTIVES: This study was done to evaluate the effectiveness and the safety of enhanced penetration of platelet rich plasma with ultrasound after plasma fractional radiofrequency for the treatment of striae distensae. SUBJECTS AND METHOD: Eighteen participants with striae distensae were treated with a Legato system (Alma Lasers, Israel) every two weeks for a total of four sessions. Thereafter, in order to enhance platelet-rich plasma penetration, ultrasound is applied. Clinical photographs were taken before first treatment and two months after the final treatment. Objective and subjective improvement scores were evaluated to demonstrate the efficacy. Abdominal skin biopsies were obtained from three individuals and histological changes were analyzed by light microscopy. RESULTS: During the two months after the last treatment, the average width of the widest striae had decreased from 0.75 to 0.27 mm. In the objective assessment, 71.9% of the participants reported ''good" or "very good'' overall improvement. In the subjective assessment, and 72.2% of the participants reported ''very satisfied" or "extremely satisfied'' with overall improvement. The only reported side effect was post-inflammatory hyperpigmentation (11.1%). CONCLUSIONS: The plasma fractional radiofrequency and transepidermal delivery of platelet-rich plasma using ultrasound is useful in the treatment of striae distensae.

Neural stimulation on human bone marrow-derived mesenchymal stem cells by extremely low frequency electromagnetic fields.

Adult stem cells are considered multipotent. Especially, human bone marrow-derived mesenchymal stem cells (hBM-MSCs) have the potential to differentiate into nerve type cells. Electromagnetic fields (EMFs) are widely distributed in the environment, and recently there have been many reports on the biological effects of EMFs. hBM-MSCs are weak and sensitive pluripotent stem cells, therefore extremely low frequency-electromagnetic fields (ELF-EMFs) could be affect the changes of biological functions within the cells. In our experiments, ELF-EMFs inhibited the growth of hBM-MSCs in 12 days exposure. Their gene level was changed and expression of the neural stem cell marker like nestin was decreased but the neural cell markers like MAP2, NEUROD1, NF-L, and Tau were induced. In immunofluorescence study, we confirmed the expression of each protein of neural cells. And also both oligodendrocyte and astrocyte related proteins like O4 and GFAP were expressed by ELF-EMFs. We suggest that EMFs can induce neural differentiation in BM-MSCs without any chemicals or differentiation factors.

Histological features and immune cell changes in skin lesions of engraftment syndrome of children undergoing hematopoietic stem cell transplantation.

Various skin eruptions are encountered during hematopoietic stem cell transplantation (HSCT) of children with hematologic malignancies. Engraftment syndrome (ES) is a disease characterized by fever, weight gain, maculopapular skin rash and noncardiogenic pulmonary edema. ES occurs during neutrophil recovery without identifiable causes of infection. Early detection of ES is critical to reduce mortality and morbidity, but identical morphologic changes found in skin lesions from ES and graft-versus-host disease (GVHD) are a challenging problem for histology-based diagnosis. To resolve this issue, immunopathologic changes in skin lesions of ES were studied. Five skin biopsies from patients with symptoms clinically compatible with ES were retrieved and compared to 15 age- and sex-matched cases of acute GVHD with antibodies to CD3, CD4, CD8 and CD1a. Mean numbers of epidermal CD8+ cells and CD1a+ cells were lower in ES than in GVHD. However, there were no significant differences in mean score of GVHD grade, mean numbers of lymphoid cells, CD3+ cells, or CD4+ cells. In the setting of HSCT in children, the dominance of CD4+ cells and a decreased number of CD1a+ cells in the epidermis are specific features for the skin lesions of ES.

Malondialdehyde inhibits an AMPK-mediated nuclear translocation and repression activity of ALDH2 in transcription.

Aging process results from deleterious damages by reactive oxygen species, in particular, various metabolic aldehydes. Aldehyde dehydrogenase 2 (ALDH2) is one of metabolic enzymes detoxifying various aldehydes under oxidative conditions. AMP-activated protein kinase (AMPK) plays a key role in controlling metabolic process. However, little was known about the relationship of ALDH2 with AMPK under oxidative conditions. Here, we, by using MDA-specific monoclonal antibody, screened the tissues of young and old rats for MDA-modified proteins and identified an ALDH2 as a prominent MDA-modified protein band in the old rat kidney tissue. ALDH2 associates with AMPK and is phosphorylated by AMPK. In addition, AICAR, an activator of AMP-activated protein kinase, induces the nuclear translocation of ALDH2. ALDH2 in nucleus is involved in general transcription repression by association with histone deacetylases. Furthermore, MDA modification inhibited the translocation of ALDH2 and the association with AMPK, and ultimately led to de-repression of transcription in the reporter system analysis. In this study, we have demonstrated that ALDH2 acts as a transcriptional repressor in response to AMPK activation, and MDA modifies ALDH2 and inhibits repressive activity of ALDH2 in general transcription. We thus suggest that increasing amount of MDA during aging process may interrupt the nuclear function of ALDH2, modulated by AMPK.

Application of mesenchymal stem cells derived from bone marrow and umbilical cord in human hair multiplication.

BACKGROUND: The methods currently used for treating alopecia have some limitations. The drug treatment is so temporary that medication discontinuance may progress depilation immediately. The number of hair transplantation restricts because total transplantable hair number is no increase. To overcome these problems, researchers have attempted the in vitro culturing of hair follicle cells and implanting these cells in the treatment area. OBJECTIVES: In the present study, culture-expanded mesenchymal stem cells (MSCs) that do not possess aggregative activity were used to produce self-aggregated cell-aggregated spheroidal dermal papilla like tissues (DPLTs) with the aid of a special culture condition in vitro, and hair bulb structure inductive capacity pertinent to the aggregative activity was then evaluated. Then hair inducing activity of self-aggregated DPLTs employing MSCs was tested in athymic mice. METHODS: We isolated and cultivated MSCs from bone marrow and umbilical cord in vitro. After propagated MSCs underwent preconditioning in dermal papilla forming medium (DPFM), then subcultured MSCs formed self-aggregated DPLTs. We compared real human scalp dermal papilla cells (hDPCs) with DPLTs employing DPCs, DPLTs employing hBM-MSCs and DPLTs employing hUC-MSCs. RESULTS: Light microscopy and immunohistochemical staining were used to confirm that reconstructed DPLTs generated by this procedure had the size, shape, and expression of protein similar to actual DP. CONCLUSIONS: The DPLTs have the same hair bulb structure inductive ability as natural DPLTs in vitro. Transplanted DPLTs can induce new hair follicle in athymic mice. As a result, UC-MSCs and BM-MSCs may be an applicable and novel cell source for the generation of human hair cell therapy.

Monoclonal antibodies to human transglutaminase 4.

Transglutaminase 4 (TG4) is a member of the enzyme family that catalyzes the calcium-dependent post-translational modification of proteins via cross-linking, polyamination, or deamidation. TG4 exhibits prostate-specific expression pattern and plays a crucial role in the formation of the copulatory plug in rodents. However, the physiological function(s) of human TG4 remains speculative. Human TG4 has been postulated to participate in the maturation process of sperm by modifying its cell surface, which results in suppression of sperm antigenicity in the female genital tract. To better understand the pathophysiological role of TG4 in prostate tissue, we generated monoclonal antibodies (MAb) against human TG4 in mice by repeated injections with the recombinant human TG4. Western blot analysis demonstrated that the selected MAbs react specifically with TG4, but not with other isoenzymes of the TG family. Immunocytochemical and immunohistochemical analyses showed that specific staining is observed with the cells overexpressing TG4 and with the paraffin-embedded prostate tissue specimens obtained from the benign prostate hyperplasia and prostate cancer patients, respectively. Our results indicate that these MAbs are suitable for detecting TG4 in the cultured cells or prostate tissues for investigating the biological functions of human TG4.

Formononetin prevents ovariectomy-induced bone loss in rats.

The major risk factor of postmenopausal osteoporosis is estrogen deficiency. Hormone replacement therapy is efficacious against osteoporosis, but it induces several significant adverse effects. In this study, therefore, we compared therapeutic potencies of three phytoestrogens: genistein, daidzein, and formononetin. Our result showed that in Saos-2 cells, formononetin and genistein (5 x 10(-7) M) treatment increased alkaline phosphatase activity by 33.0 +/- 5.8% and 21.1 +/- 4.0%. Genistein inhibited osteoclast formation in a dose-dependent manner. In OVX rats, formononetin-treated groups given 1 and 10 mg/kg/day displayed increased trabecular bone areas (TBAs) within the tibia. Genistein- and daidzein-treated groups also displayed increased tibial TBAs. TBAs of the lumbar vertebrae were higher in all treated groups than in the control group. In conclusion, formononetin as well as other isoflavones, such as daidzein and genistein, inhibited bone loss caused by estrogen-deficiency.

Differential alternative splicing of human transglutaminase 4 in benign prostate hyperplasia and prostate cancer.

Transglutaminase 4 is a member of enzyme family that catalyzes calcium-dependent posttranslational modification of proteins. Although transglutaminase 4 has been shown to have prostate-restricted expression pattern, little is known about the biological function of transglutaminase 4 in human. To gain insight into its role in prostate, we analyzed the expression status of human transglutaminase 4 in benign prostate hyperplasia (BPH) and prostate cancer (PCa). Unexpectedly, RT-PCR and nucleotide sequence analysis showed four alternative splicing variants of transglutaminase 4: transglutaminase 4-L, -M (-M1 and -M2) and -S. The difference between transglutaminase 4-M1 and -M2 is attributed to splicing sites, but not nucleotide size. The deduced amino acid sequences showed that transglutaminase 4-L, -M1 and -M2 have correct open reading frames, whereas transglutaminase 4-S has a truncated reading frame. RT-PCR analysis of clinical samples revealed that transglutaminase 4-M and -S were detected in all tested prostate tissue (80 BPH and 48 PCa). Interestingly, transglutaminase 4-L was found in 56% of BPH (45 out of 80) and only in 15% of PCa (7 out of 48). However, transglutaminase 4-L expression did not correlate with serum prostate-specific antigen (PSA) level, prostate volumes or PSA densities. These results will provide a clue to future investigation aiming at delineating physiological and pathological roles of human transglutaminase 4.

Pretibial epidermolysis bullosa: is this case a new subtype with loss of types IV and VII collagen?

Pretibial epidermolysis bullosa (PEB) is an extremely rare subtype of dominant dystrophic epidermolysis bullosa (DDEB), in which recurrent blistering with scarring predominantly involves the pretibial skin. Nail dystrophy, albopapuloid lesions, and hypertrophic scars may also occur. In PEB, immunohistochemical and electron microscopic studies demonstrate the complete or partial loss of the anchoring fibril (AF) in the basement membrane zone, suggesting disturbed synthesis or excessive degradation of collagen VII, the main component of AF. Interestingly, we report a case of PEB with unusual results of joint loss of types IV and VII collagen.

Increase in cell migration and angiogenesis in a composite silk scaffold for tissue-engineered ligaments.

The purpose of this study was to evaluate the biocompatibility of silk and collagen-hyaluronan (HA) in vitro by assessing anterior cruciate ligament (ACL) cell and T-lymphocyte cultures on scaffolds. The use of composite scaffolds as artificial ligaments in ACL reconstruction and their effects on angiogenesis were evaluated in vivo. The silk scaffold was knitted by hand and dry coated with collagen-HA, whereas the composite silk scaffold was made by covering a silk scaffold with a lyophilized collagen-HA substrate. The initial attachment and proliferation of human ACL cells on the composite silk scaffold was superior to the attachment and proliferation observed on the silk scaffold. The immune response was higher in both scaffolds after 72 h (p < 0.05) compared with the control culture condition without scaffolding, as assessed by T-lymphocyte cultures in vitro. There was no significant difference in the immune response in vitro between the silk and composite silk scaffolds. Silk and composite silk scaffolds were implanted as artificial ligaments in ACLs removed from the knees of dogs, and they were harvested 6 weeks after implantation. On gross examination, the onset of an inflammatory tissue reaction, such as synovitis, was seen in both the silk scaffold and the composite silk scaffold groups. An histological evaluation of the artificial ligament implants revealed the presence of monocytes in the silk composite scaffold and the absence of giant cells in all cases. MT staining in the composite silk scaffold-grafted group showed granulation tissue consisting of fibroblasts, lymphocytes, monocytes, and collagen fibers. In addition, CD31 staining revealed the formation of new blood vessels. On the other hand, no reparative tissues, such as blood vessels, collagen, and cells, were observed in the silk scaffold-grafted group. These results suggest that the lyophilized collagen-HA substrate is biocompatible in vitro and enhances new blood vessel and cell migration in vivo.

A case of diffuse neurofibroma of the scalp.

A neurofibroma is a benign tumor of the peripheral nerve sheath characterized by proliferation of Schwann cells, perineural cells, and endoneurial fibroblasts. Different types of neurofibromas can be identified, including localized, plexiform, and diffuse types. Neurofibromas can involve any site on the body skin. The diffuse variant is rare and occurs primarily in children and young adults. It involves the skin and subcutaneous tissue in a plaque-like fashion on the head and neck regions. We present a case of a 10-year-old boy who had a diffuse neurofibroma on the scalp.

Lysophosphatidic acid and adenylyl cyclase inhibitor increase proliferation of senescent human diploid fibroblasts by inhibiting adenosine monophosphate-activated protein kinase.

This study was designed to elucidate the molecular mechanism underlying lysophosphatidic acid (LPA) and adenylyl cyclase inhibitor SQ22536 (ACI)-induced senescent human diploid fibroblast (HDF) proliferation. Because adenosine monophosphate (AMP)-activated protein kinase (AMPK) is known to inhibit cell proliferation, we examined the phosphorylation status of AMPK and p53 and the expression level of p21(waf1/cip1) after treating HDFs with LPA and ACI. Phosphorylation of AMPKalpha on threonine-172 (p-Thr172-AMPKalpha) increases its catalytic activity but phosphorylation on serine-485/491 (p-Ser485/491-AMPKalpha) reduces the accessibility of the Thr172 phosphorylation site thereby inhibiting its catalytic activity. LPA increased p-Ser485/491-AMPKalpha, presumably by activating cAMP-dependent protein kinase (PKA). However, ACI reduced p-Thr172-AMPKalpha by inhibiting the LKB signaling. Our data demonstrated that both LPA and ACI inhibit the catalytic activity of AMPKalpha and p53 by differentially regulating phosphorylation of AMPKalpha, causing increased senescent cell proliferation. These findings suggest that the proliferation potential of senescent HDFs can be modulated through the regulation of the AMPK signaling pathway.