DERMATOLOGIC SURGERY, Issue 6 2006
NATALIA CYMROT CYMBALISTA MD
BACKGROUND Idiopathic cutaneous hyperchromia of the orbital region (ICHOR) does not have a clear etiopathogenesis. Genetic factors, increased melanin, prominent vasculature, and eyelid skin slackness seem to be involved. OBJECTIVE To evaluate individuals with ICHOR clinically and histologically, before and after treatment with high-energy pulsed light (HEPL), considering epidermal and dermal melanin, in order to evaluate HEPL efficacy in clearing away ICHOR, and 1 month and 1 year later to check whether improvement was maintained. METHODS Twelve individuals with ICHOR underwent clinical and histological evaluation before and after HEPL application, with photographic comparison. They underwent one to four HEPL sessions on the lower eyelid at approximately 30-day intervals. Melanin quantification by area, before and after treatment was performed by digital image morphometry. RESULTS Eyelid skin was significantly lightened (p=.24), and was maintained 1 year later with no ICHOR reincidence. All individuals (100%) showed postinflammatory hyperchromia (average 6-month duration), while 58.33% presented hypochromia (7-month duration). There was significantly decreased epidermal and dermal melanin after treatment. CONCLUSION HEPL was shown to be useful in clearing up ICHOR. This was maintained after 1 year. Epidermal and dermal histopathology showed decreased melanin following treatment. Longer follow-up is needed to evaluate possible later recurrence of ICHOR. [source]

Glycolic Acid Treatment Increases Type I Collagen mRNA and Hyaluronic Acid Content of Human Skin

DERMATOLOGIC SURGERY, Issue 5 2001
Eric F. Bernstein MD
Background. Chronic solar irradiation results in both morphologic and functional changes in affected skin. ,-hydroxy acids, such as glycolic acid, have been shown to improve photodamaged skin. Objective. To investigate alterations in collagen gene induction and epidermal and dermal hyaluronic acid production as a result of administered glycolic acid. Methods. In this study we compared collagen gene expression from skin biopsy specimens, and epidermal and dermal hyaluronic acid immunohistochemical staining between glycolic acid-treated and vehicle-treated skin. Forearm skin was treated with 20% glycolic acid lotion or a lotion vehicle control twice a day for 3 months. Results. Epidermal and dermal hyaluronic acid and collagen gene expression were all increased in glycolic acid-treated skin as compared to vehicle-treated controls. Conclusion. Our data suggest that epidermal and dermal remodeling of the extracellular matrix results from glycolic acid treatment. Longer treatment intervals may result in collagen deposition as suggested by the measured increase in mRNA. [source]

Sodium lauryl sulphate alters the mRNA expression of lipid-metabolizing enzymes and PPAR signalling in normal human skin in vivo

EXPERIMENTAL DERMATOLOGY, Issue 12 2009
Hans Törmä
Abstract:, Detergents irritate skin and affect skin barrier homeostasis. In this study, healthy skin was exposed to 1% sodium lauryl sulphate (SLS) in water for 24 h. Biopsies were taken 6 h to 8 days post exposure. Lipid patterns were stained in situ and real-time polymerase chain reaction (PCR) was used to examine mRNA expression of enzymes synthesizing barrier lipids, peroxisome proliferator-activated receptors (PPAR) and lipoxygenases. The lipid pattern was disorganized from 6 h to 3 days after SLS exposure. Concomitant changes in mRNA expression included: (i) reduction, followed by induction, of ceramide-generating ,-glucocerebrosidase, (ii) increase on day 1 of two other enzymes for ceramide biosynthesis and (iii) persistent reduction of acetyl-CoA carboxylase-B, a key enzyme in fatty acid synthesis. Surprisingly, the rate-limiting enzyme in cholesterol synthesis, HMG-CoA reductase, was unaltered. Among putative regulators of barrier lipids synthesis, PPAR, and PPAR, exhibited reduced mRNA expression, while PPAR,/, and LXR, were unaltered. Epidermal lipoxygenase-3, which may generate PPAR, agonists, exhibited reduced expression. In conclusion, SLS induces reorganization of lipids in the stratum corneum, which play a role in detergents' destruction of the barrier. The changes in mRNA expression of enzymes involved in synthesizing barrier lipids are probably important for the restoration of the barrier. [source]

Two Japanese cases of lichen planus pigmentosus-inversus

INTERNATIONAL JOURNAL OF DERMATOLOGY, Issue 7 2007
Aki Kashima MD
Case 1 was a 51-year-old Japanese woman. She presented with an asymptomatic brown macule located on the right axilla of 2 months' duration. The smooth macule was 2 cm in diameter with a sharp demarcation (Fig. 1A). Figure 1. Photographs of skin lesions in two patients. (A) Case 1. Well-circumscribed brown macule without an active red border in the central portion of the right axilla. (B) Case 2. Symmetric distribution of brown macules without an active red border in the popliteal fossae Case 2 was a 62-year-old Japanese man. He presented with asymptomatic, symmetric, gray,brown macules located on the groin, axillae, and popliteal region of 6 months' duration. The smooth macules were several millimeters to centimeters in diameter and sharply demarcated (Fig. 1B). Oral or nail lesions, previous inflammatory processes in affected areas, and internal malignancies were absent. A causal relationship with drugs, recent sun exposure, or trauma could not be identified. Findings for work-up, including blood cell count, fasting blood sugar levels, liver function, serum electrolyte levels, serum electrophoresis, urinalysis, antinuclear antibodies, and serological examinations for human hepatitis viruses and syphilis, were within normal limits or negative. The lesions gradually disappeared without medication within 6 months. Biopsy specimens showed a lymphocytic infiltrate with basal vacuolar changes and prominent melanin incontinence in the upper dermis (Fig. 2A). The band-like lymphocytic infiltrate was moderate in Case 1 and mild in Case 2. Immunohistochemistry showed infiltrative CD8+ T lymphocytes with keratinocytic damage, indicating cytotoxic injury of the keratinocytes (Fig. 2B). Both the epidermis and the upper dermis contained CD1a+ cells (Fig. 2C). The keratinocytes focally and weakly expressed HLA-DR (Fig. 2D). These findings were identical in samples from both patients. Figure 2. Light and immunohistochemical microphotographs. (A) Mild, band-like, lymphocytic infiltrate with basal vacuolar change and prominent melanin incontinence in the upper dermis with apoptosis or necrosis of keratinocytes. (B) Epidermal infiltrate of CD8+ T lymphocytes with keratinocytic damage. (C) CD1a+ cells in the upper dermis. (D) Keratinocytes focally and weakly express HLA-DR (original magnifications: A, ×200; B,D, ×400) [source]

Improvement of port wine stain laser therapy by skin preheating prior to cryogen spray cooling: A numerical simulation

LASERS IN SURGERY AND MEDICINE, Issue 2 2006
Wangcun Jia PhD
Abstract Background and Objectives Although cryogen spray cooling (CSC) in conjunction with laser therapy has become the clinical standard for treatment of port wine stain (PWS) birthmarks, the current approach does not produce complete lesion blanching in the vast majority of patients. The objectives of this study are to: (1) experimentally determine the dynamic CSC heat flux when a skin phantom is preheated, and (2) numerically study the feasibility of using skin preheating prior to CSC to improve PWS laser therapeutic outcome. Study Design/Materials and Methods A fast-response thin-foil thermocouple was used to measure the surface temperature and thus heat flux of an epoxy skin phantom during CSC. Using the heat flux as a boundary condition, PWS laser therapy was simulated with finite element heat diffusion and Monte Carlo light distribution models. Epidermal and PWS blood vessel thermal damage were calculated with an Arrhenius-type kinetic model. Results Experimental results show that the skin phantom surface can be cooled to a similar minimum temperature regardless of the initial temperature. Numerical simulation indicates that upon laser irradiation, the epidermal temperature increase is virtually unaffected by preheating, while higher PWS blood vessel temperatures can be achieved. Based on the damage criterion we assumed, the depth and maximum diameter of PWS vessels that can be destroyed irreversibly with skin preheating are greater than those without. Conclusions Skin preheating prior to CSC can maintain epidermal cooling while increasing PWS blood vessel temperature before laser irradiation. Numerical models have been developed to show that patients may benefit from the skin preheating approach, depending on PWS vessel diameter and depth. Lasers Surg. Med. 38:155,162, 2006. © 2006 Wiley-Liss, Inc. [source]

DNA vaccination against tumors

THE JOURNAL OF GENE MEDICINE, Issue 1 2005
Gérald J. Prud'homme
Abstract DNA vaccines have been used to generate protective immunity against tumors in a variety of experimental models. The favorite target antigens have been those that are frequently expressed by human tumors, such as carcinoembryonic antigen (CEA), ErbB2/neu, and melanoma-associated antigens. DNA vaccines have the advantage of being simple to construct, produce and deliver. They can activate all arms of the immune system, and allow substantial flexibility in modifying the type of immune response generated through codelivery of cytokine genes. DNA vaccines can be applied by intramuscular, dermal/epidermal, oral, respiratory and other routes, and pose relatively few safety concerns. Compared to other nucleic acid vectors, they are usually devoid of viral or bacterial antigens and can be designed to deliver only the target tumor antigen(s). This is likely to be important when priming a response against weak tumor antigens. DNA vaccines have been more effective in rodents than in larger mammals or humans. However, a large number of methods that might be applied clinically have been shown to ameliorate these vaccines. This includes in vivo electroporation, and/or inclusion of various immunostimulatory molecules, xenoantigens (or their epitopes), antigen-cytokine fusion genes, agents that improve antigen uptake or presentation, and molecules that activate innate immunity mechanisms. In addition, CpG motifs carried by plasmids can overcome the negative effects of regulatory T cells. There have been few studies in humans, but recent clinical trials suggest that plasmid/virus, or plasmid/antigen-adjuvant, prime-boost strategies generate strong immune responses, and confirm the usefulness of plasmid-based vaccination. Copyright © 2004 John Wiley & Sons, Ltd. [source]

Effect of Laser Resurfacing on p53 Expression in Photoaged Facial Skin

DERMATOLOGIC SURGERY, Issue 6 2007
MOETAZ M. EL-DOMYATI MD
BACKGROUND p53 overexpression has been reported in photoaged skin. Meanwhile, p53 gene mutations have been implicated as an important factor in the pathogenesis of ultraviolet (UV) light,induced skin cancer. OBJECTIVE The objective was to evaluate the effect of laser resurfacing on the epidermal thickness and expression of p53 in photoaged skin. METHODS Specimens were obtained from the facial skin of 10 patients before and after 3 months and 1 year of treatment using CO2 (five cases) and erbium (Er):YAG (five cases) lasers. Specimens were also obtained from six age-matched controls. These biopsies were used for routine histopathology, histometry, and p53 immunoperoxidase staining. RESULTS Both CO2 and Er:YAG lasers were found to induce a significant decrease in p53 expression in biopsies obtained after 3 months (p=.0004 and .002, respectively) followed by gradual increase (p=.01 in both groups). A significant increase (p<.01) in epidermal thickness was also observed after 1 year of resurfacing. This increase, however, is inversely correlated with the level of p53 expression in such patients. CONCLUSION The decrease in epidermal p53 expression after CO2 and Er:YAG lasers may account for some of the benefits of resurfacing on the epidermis, as well as prevention of actinic neoplasia by adjusting any disturbance in the proliferation/apoptosis balance observed in photoaged facial skin. [source]

Histometric and Histochemical Analysis of the Effect of Trichloroacetic Acid Concentration in the Chemical Reconstruction of Skin Scars Method

DERMATOLOGIC SURGERY, Issue 10 2006
SUNG BIN CHO MD
BACKGROUND Atrophic scars can be induced by various causes, including severely inflamed acne, chicken pox, and trauma. Many treatment modalities are used for reconstructing and improving the appearance of scars with various treatment results. OBJECTIVE A recent report shows the clinical efficacy of the chemical reconstruction of skin scars (CROSS) method, which consists of the focal application of trichloroacetic acid (TCA) in a higher concentration. Histometric analysis of the CROSS method, however, has not yet been established. METHODS In this study, five hairless mice were used to evaluate the effect of the CROSS method and to analyze the difference between the CROSS method and simple TCA application. RESULTS Similar histologic changes were observed in the two methods, including epidermal and dermal rejuvenation with new collagen deposition. These changes, however, were more prominent in the CROSS method,treated areas, particularly when 100% TCA was used. CONCLUSION The results of this study suggest that treatment of atrophic scars using the CROSS method is more effective than simple application of TCA in activating fibroblasts in the dermis and increasing the amount of collagen. [source]

Treatment of Idiopathic Cutaneous Hyperchromia of the Orbital Region (ICHOR) with Intense Pulsed Light

Sebaceous Carcinoma, Basal Cell Carcinoma, Trichoadenoma, Trichoblastoma, and Syringocystadenoma Papilliferum Arising within a Nevus Sebaceus

DERMATOLOGIC SURGERY, Issue 12p2 2004
Christopher J. Miller MD
Background. Nevus sebaceus has a well-documented potential to develop a wide variety of neoplasms of both epidermal and adnexal origins. It is highly unusual for more than three tumors to arise simultaneously within a single nevus sebaceus. Sebaceous carcinoma arising within a nevus sebaceus is a rare occurrence. Objective. The objective was to report the case of a patient with a nevus sebaceus that simultaneously developed five distinct neoplasms of epidermal and various adnexal origins and to report the fourth case of sebaceous carcinoma arising within a nevus sebaceus. Methods. A 45-year-old woman presented with a nevus sebaceusthat contained five separate neoplasms, including sebaceous carcinoma, basal cell carcinoma, trichoadenoma, trichoblastoma, and syringocystadenoma papilliferum. Results. Complete excision of the nevus sebaceus and the five tumors was performed. Systemic work-up showed no evidence of metastatic disease or association with Muir-Torre syndrome. Conclusion. This case report highlights the diverse neoplastic potential of nevus sebaceus and demonstrates the capacity of this hamartoma to develop aggressive tumors, such as sebaceous carcinoma. Prophylactic excision or at least close clinical surveillance for sudden development of new growths is warranted in all cases of nevus sebaceus. [source]

Combined Therapy Using Q-Switched Ruby Laser and Bleaching Treatment With Tretinoin and Hydroquinone for Acquired Dermal Melanocytosis

DERMATOLOGIC SURGERY, Issue 10 2003
Akira Momosawa MD
Background and objective. Acquired dermal melanocytosis (ADM; acquired bilateral nevus of Ota-like macules) is known for its recalcitrance compared with Nevus of Ota, and we assume that one of the reasons is a higher rate and degree of postinflammatory hyperpigmentation (PIH) seen after laser treatments. Methods. Topical bleaching treatment with 0.1% tretinoin aqueous gel and 5% hydroquinone ointment containing 7% lactic acid was initially performed (4 to 6 weeks) to discharge epidermal melanin. Subsequently, Q-switched ruby (QSR) laser was irradiated to eliminate dermal pigmentation. Both steps were repeated two to three times until patient satisfaction was obtained (usually at a 2-month interval for laser sessions). This treatment was performed in 19 patients with ADM. Skin biopsy was performed in six cases at baseline, after the bleaching pretreatment, and at the end of treatment. Results. All patients showed good to excellent clearing after two to three sessions of QSR laser treatments. The total treatment period ranged from 3 to 13 (mean of 8.3) months. PIH was observed in 10.5% of the cases. Histologically, epidermal hyperpigmentation was observed in all specimens and was dramatically improved by the topical bleaching pretreatment. Conclusion. QSR laser combined with the topical bleaching pretreatment appeared to treat ADM consistently with a low occurrence rate of PIH and lessen the number of laser sessions and total treatment period and may also be applied to any other lesions with both epidermal and dermal pigmentation. [source]

Glycolic Acid Treatment Increases Type I Collagen mRNA and Hyaluronic Acid Content of Human Skin

Functional analysis of synaptotagmin gene regulatory regions in two distantly related ascidian species

DEVELOPMENT GROWTH & DIFFERENTIATION, Issue 7 2008
Jun Matsumoto
We have studied the structure and function of a promoter region of the Halocynthia synaptotagmin (Hr-Syt) gene, which is abundantly expressed in neuronal cells. Our previous analysis suggested that the expression of Hr-Syt is regulated by at least one epidermal and two neuronal regulatory regions. In this study, the regulatory regions of Hr-Syt promoter were further characterized by using two species of ascidians, Halocynthia roretzi and Ciona intestinalis embryos. A putative GATA transcription factor binding site in the epidermal regulatory region has ectodermal enhancer activity in the Halocynthia embryo. Neuronal expression of Hr-Syt was regulated by multiple redundant enhancer regions. Among these enhancer regions, a 200-bp (,2900/,2700) region drove the reporter expression in neurons in both species of ascidian. Although the synaptotagmin promoter sequences did not show overall similarity between Hr-Syt and Ciona synaptotagmin (Ci-Syt), 5,-upsteream two short sequences of Ci-Syt have similarity to the ,2766/,2732 region of the Hr-Syt promoter. The homeodomain binding sites in this region are required for the neuronal enhancer activity. These results suggest that GATA and homeodomain transcription factors regulate the expression of synaptotagmin. [source]

Epidermal transient down-regulation of retinol - binding protein 4 and mirror expression of apolipoprotein Eb and estrogen receptor 2a during zebrafish fin and scale development

DEVELOPMENTAL DYNAMICS, Issue 11 2006
Angèle Tingaud-Sequeira
Abstract Very little is known about the molecular control of skin patterning and scale morphogenesis in teleost fish. We have found radially symmetrical epidermal placodes with down-regulation of retinol - binding protein 4 (rbp4) expression during the initial paired fin and scale morphogenesis in zebrafish (Danio rerio). This finding may be related to changes in keratinocyte cytodifferentiation and/or the integument retinoid metabolism. rbp4 transcripts are expressed afterward in the central epidermis of the scale papilla and gradually extend to the epidermis, covering the growing scale, whereas no transcripts were detected in posterior margin epidermis. In contrast, induction of apolipoprotein Eb (apoeb) and up-regulation of estrogen receptor 2a (esr2a) transcripts were observed in the epidermis at initiator sites of zebrafish ectodermal/dermal appendage morphogenesis. This expression was maintained in the posterior margin epidermis of the formed scales. esr2a was also strongly expressed in neuromasts, whereas no rbp4 and apoeb transcripts were detected in these mechanosensory structures. The observed epidermal molecular events suggest that epidermis patterning is due to an activator,inhibitor mechanism operational at epidermal,dermal interaction sites. rbp4 transcript expression was also strongly down-regulated by 1-phenyl-2-thio-urea (PTU). As this inhibitor is commonly used to block obscuring pigmentation during in situ hybridization studies, this finding suggests that PTU should be used with caution, particularly in studying skin development. Developmental Dynamics 235:3071,3079, 2006. © 2006 Wiley-Liss, Inc. [source]

Conserved RARE localization in amphioxus Hox clusters and implications for Hox code evolution in the vertebrate neural crest

DEVELOPMENTAL DYNAMICS, Issue 6 2006
Hiroshi Wada
Abstract The Hox code in the neural crest cells plays an important role in the development of the complex craniofacial structures that are characteristic of vertebrates. Previously, 3, AmphiHox1 flanking region has been shown to drive gene expression in neural tubes and neural crest cells in a retinoic acid (RA)-dependent manner. In the present study, we found that the DR5-type RA response elements located at the 3, AmphiHox1 flanking region of Branchiostoma floridae are necessary and sufficient to express reporter genes in both the neural tube and neural crest cells of chick embryos, specifically at the post-otic level. The DR5 at the 3, flanking region of chick Hoxb1 is also capable of driving the same expression in chick embryos. We found that AmphiHox3 possesses a DR5-type RARE in its 5, flanking region, and this drives an expression pattern similar to the RARE element found in the 3, flanking region of AmphiHox1. Therefore, the location of these DR5-type RAREs is conserved in amphioxus and vertebrate Hox clusters. Our findings demonstrate that conserved RAREs mediate RA-dependent regulation of Hox genes in amphioxus and vertebrates, and in vertebrates this drives expression of Hox genes in both neural crest and neural tube. This suggests that Hox expression in vertebrate neural crest cells has evolved via the co-option of a pre-existing regulatory pathway that primitively regulated neural tube (and possibly epidermal) Hox expression. Developmental Dynamics 235:1522,1531, 2006. © 2006 Wiley-Liss, Inc. [source]

Cx31 and Cx43 double-deficient mice reveal independent functions in murine placental and skin development

DEVELOPMENTAL DYNAMICS, Issue 3 2005
Mark Kibschull
Abstract The overlapping expression of gap junctional connexins in tissues has indicated that the channels may compensate for each other. During development, Cx31 and Cx43 are coexpressed in preimplantation embryos, in the spongiotrophoblast of the placenta and in the epidermis. This study shows that Cx31/Cx43 double-deficient mice exhibit the known phenotypes of the single-knockout strains but no combined effects. Thus, Cx43, coexpressed with Cx31 at midgestation in the spongiotrophoblast of the placenta, cannot be responsible for a partial rescue of the lethal Cx31 knockout phenotype, as assumed before (Plum et al. [ 2001] Dev Biol 231:334,337). It follows that both connexins have unique functions in placental development. Despite an altered expression of other epidermal connexin mRNAs, epidermal differentiation and physiology was unaltered by the absence of Cx31 and Cx43. Therefore, in epidermal and preimplantation development, gap junctional communication can probably be compensated by other isoforms coexpressed with Cx31 and Cx43. Developmental Dynamics 233:853,863, 2005. © 2005 Wiley-Liss, Inc. [source]

Retinoids and nonvertebrate chordate development

DEVELOPMENTAL NEUROBIOLOGY, Issue 7 2006
Shigeki Fujiwara
Abstract Retinoic acid (RA) is required for the differentiation and morphogenesis of chordate-specific features, such as the antero-posterior regionalization of the dorsal hollow nerve cord and neural crest cells. RA receptors (RARs) have been reported exclusively in chordates, suggesting that the acquisition of the RAR gene was important for chordate evolution. A scenario is presented here for the establishment of an RAR-mediated developmental regulatory system during the course of chordate evolution. In the common chordate ancestor, RAR came to control the spatial expression pattern of Hox genes in the ectoderm and endoderm along the antero-posterior axis. In these germ layers, RA was required for the differentiation of epidermal sensory neurons and the morphogenesis of pharyngeal gill slits, respectively. As the diffuse epidermal nerve net in the chordate ancestor became centralized to form the dorsal nerve cord, the epidermal Hox expression pattern was carried into the central nervous system. Because the Hox code here came to specify neuronal identity along the antero-posterior axis, RA became inextricably linked to the antero-posterior patterning of the chordate central nervous system. © 2006 Wiley Periodicals, Inc. J Neurobiol 66: 645,652, 2006 [source]

The role of neuropeptides and neuropeptide-degrading enzymes in wound healing

EXPERIMENTAL DERMATOLOGY, Issue 9 2004
John E. Olerud
Thirty to 40% of diabetic patients develop sensory neuropathy. Neuropathy is a major causal factor in diabetic ulcers. Only 31% of neuropathic diabetic ulcers heal in 20 weeks. Patients with neuropathy have a 15.5-fold excess risk of amputation. Diabetic patients with neuropathy particularly lose epidermal and papillary dermal sensory nerves which release neuropeptides such as substance P (SP). Neutral endopeptidase (NEP), the enzyme that degrades SP, is dramatically over expressed in patients with diabetic neuropathy. SP has positive effect on wound healing. Treatment strategies related to the nervous system for prevention and treatment of diabetic ulcers currently being studied include prevention of neuropathy with tight control of blood glucose, application of neuropeptides, nerve growth factors (NGF), and antagonists of NEP. [source]

Cloning, characterization and localization of a novel basic peroxidase gene from Catharanthus roseus

FEBS JOURNAL, Issue 5 2007
Santosh Kumar
Catharanthus roseus (L.) G. Don produces a number of biologically active terpenoid indole alkaloids via a complex terpenoid indole alkaloid biosynthetic pathway. The final dimerization step of this pathway, leading to the synthesis of a dimeric alkaloid, vinblastine, was demonstrated to be catalyzed by a basic peroxidase. However, reports of the gene encoding this enzyme are scarce for C. roseus. We report here for the first time the cloning, characterization and localization of a novel basic peroxidase, CrPrx, from C. roseus. A 394 bp partial peroxidase cDNA (CrInt1) was initially amplified from the internodal stem tissue, using degenerate oligonucleotide primers, and cloned. The full-length coding region of CrPrx cDNA was isolated by screening a leaf-specific cDNA library with CrInt1 as probe. The CrPrx nucleotide sequence encodes a deduced translation product of 330 amino acids with a 21 amino acid signal peptide, suggesting that CrPrx is secretory in nature. The molecular mass of this unprocessed and unmodified deduced protein is estimated to be 37.43 kDa, and the pI value is 8.68. CrPrx was found to belong to a ,three intron' category of gene that encodes a class III basic secretory peroxidase. CrPrx protein and mRNA were found to be present in specific organs and were regulated by different stress treatments. Using a ,-glucuronidase,green fluorescent protein fusion of CrPrx protein, we demonstrated that the fused protein is localized in leaf epidermal and guard cell walls of transiently transformed tobacco. We propose that CrPrx is involved in cell wall synthesis, and also that the gene is induced under methyl jasmonate treatment. Its potential involvement in the terpenoid indole alkaloid biosynthetic pathway is discussed. [source]

Prediction of jet flows in the supersonic nozzle and diffuser

INTERNATIONAL JOURNAL FOR NUMERICAL METHODS IN FLUIDS, Issue 10-11 2005
Yi Liu
Abstract The authors' recently-developed code for a needle-free powdered vaccine delivery device, the epidermal powdered inject system (EPI), is summarized in this paper. The behaviour of supersonic jet flows, which accelerate micron sized powdered vaccines to penetrate human skin or mucosal tissue, is therefore of great importance. A well-established modified implicit flux vector splitting (MIFVS) solver for the Navier,Stokes equations is extended to study numerically the transient supersonic jet flows of interest. A low Reynolds number k,, turbulence model, with the compressibility effect considered, is integrated into MIFVS solver to predict the turbulent structures and interactions with inherent shock systems. The results for the NASA validation case NPARC, Venturi and contoured shock tube (CST) of the EPI system are discussed. Copyright © 2005 John Wiley & Sons, Ltd. [source]

Quantitative model of cellulite: three-dimensional skin surface topography, biophysical characterization, and relationship to human perception

INTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 4 2005
L. K. Smalls
Gynoid lipodystrophy (cellulite) is the irregular, dimpled skin surface of the thighs, abdomen, and buttocks in 85% of post-adolescent women. The distinctive surface morphology is believed to result when subcutaneous adipose tissue protrudes into the lower reticular dermis, thereby creating irregularities at the surface. The biomechanical properties of epidermal and dermal tissue may also influence severity. Cellulite-affected thigh sites were measured in 51 females with varying degrees of cellulite, in 11 non-cellulite controls, and in 10 male controls. A non-contact high-resolution three-dimensional (3D) laser surface scanner was used to quantify the skin surface morphology and determine specific roughness values. The scans were evaluated by experts and na,ve judges (n = 62). Body composition was evaluated via dual-energy X-ray absorptiometry; dermal thickness and the dermal,subcutaneous junction were evaluated via high-resolution 3D ultrasound and surface photography under compression. Biomechanical properties were also measured. The roughness parameters Svm (mean depth of the lowest valleys) and Sdr (ratio between the roughness surface area and the area of the xy plane) were highly correlated to the expert image grades and, therefore, designated as the quantitative measures of cellulite severity. The strength of the correlations among na,ve grades, expert grades, and roughness values confirmed that the data quantitatively evaluate the human perception of cellulite. Cellulite severity was correlated to BMI, thigh circumference, percent thigh fat, architecture of the dermal,subcutaneous border (ultrasound surface area, red-band SD from compressed images), compliance, and stiffness (negative correlation). Cellulite severity was predicted by the percent fat and the area of the dermal,subcutaneous border. The biomechanical properties did not significantly contribute to the prediction. Comparison of the parameters for females and males further suggests that percent thigh fat and surface area roughness deviation are the distinguishing features of cellulite. [source]

An optimized method for intensive screening of molecules that stimulate , -defensin 2 or 3 (hBD2 or hBD3) expression in cultured normal human keratinocytes

INTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 3 2005
I. Pernet
Synopsis Normal human skin controls the intrusion of microorganisms by the production of peptide antibiotics such as defensins. The aim of our study was to develop a culture model of normal human keratinocytes for optimal , -defensin mRNA detection which allows the screening of molecules able to stimulate hBD2 and hBD3 without inducing pro-inflammatory cytokines. A keratinocyte culture model in 96-well plates, in high calcium medium (1.7 mm) allowed to analyze hBD2 and hBD3 mRNA expression in basal condition and after cell stimulation by products from diverse vegetal extracts. The release of IL-8 and the chemokine MIP-3, was also evaluated in cell supernatants by ELISA. Among the 184 extracts tested, 75 showed a stimulatory effect on , -defensin expression: 40 on hBD2, 26 on hBD3 and nine on both defensins. Fifteen of these substances which also induced the release of pro-inflammatory cytokines were eliminated. Among the other substances, four were selected and were analyzed in a dose-dependent study (n = 4) by real-time quantitative RT-PCR and completed by a measure of MIP-3,, IL-8 and IL-1, levels. These data underline the important necessity of screening result controls by a quantitative method reproduced at least three times. This new method of intensive screening allowed us to exhibit vegetal extracts that were able to stimulate epidermal , -defensin expression without inducing an up-secretion of pro-inflammatory cytokines. Résumé La peau humaine normale exerce une fonction barrière contre l'intrusion de microorganismes par la production de peptides antibiotiques comme les défensines. Le but de cette étude a consistéà mettre au point un modèle de culture de kératinocytes humains normaux permettant une détection optimale des ARNm des défensines en général, et adapté au screening de molécules aptes à stimuler les défensines épidermiques hBD2 et hBD3 en particulier, sans induire de cytokines pro-inflammatoires. Un modèle de culture de kératinocytes en plaques 96 puits, en milieu riche en calcium (1,7 mm) permet une analyse de l'expression des ARNm de hBD2 et hBD3 en condition basale et après stimulation par divers extraits végétaux. La sécrétion d'IL-8 et de la chimiokine MIP-3, a étéévaluée dans les surnageants de culture par ELISA. Parmi les 184 extraits testés, 75 montrent un effet stimulant sur l'expression des , -défensines : 40 ont un effet sur hBD2, 26 sur hBD3 et 9 sur les 2 types de défensines. Quinze de ces actifs qui induisent aussi la sécrétion de cytokines pro-inflammatoires ont étééliminés. Parmi les autres molécules, 4 ont été sélectionnées pour faire l'objet d'une étude de leurs effets-doses (n = 4) sur l'expression des , -défensines par une technique quantitative de RT-PCR en temps réel. Cette étude est complétée par le dosage des cytokines IL-1,, IL-8 et MIP-3,. Les résultats obtenus soulignent l'importante nécessitée de contrôler au moins trois fois par une méthode quantitative les résultats d'un screening. Cette nouvelle méthode de screening intensif nous a permis de mettre en évidence des extraits végétaux capables de stimuler les défensines épidermiques sans induire de cytokines pro-inflammatoires. [source]

Reduced barrier efficiency in axillary stratum corneum

INTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 3 2002
A. Watkinson
Synopsis The skin of the axilla is cosmetically important with millions of consumers daily applying antiperspirant/deodorant products. Despite this, we know virtually nothing about axillary skin or how antiperspirant (AP) use impacts upon it. To characterize the axillary stratum corneum and determine whether this is a unique skin type, we have looked at stratum corneum composition and function, particularly its barrier properties, and compared it with other body sites. Transepidermal water loss (TEWL) and corneosurfametry (CSM) revealed a reduced barrier function in the axilla. HPTLC analysis of the stratum corneum lipids demonstrated statistically elevated levels of fatty acids, ceramides, and particularly cholesterol in the axilla. Both ceramide and cholesterol did not appear to change with depth, indicating that they were predominantly of stratum corneum origin. On the other hand, at least some of the fatty acid had a sebaceous origin. We hypothesized that the reduced barrier function might be owing to the changes in the crucial ceramide : cholesterol ratio. To address this, we used a combination of attenuated total reflectance,Fourier-transformed infrared spectroscopy (ATR,FTIR) with cyanoacrylate sampling. These results demonstrated more ordered lipid-lamellae phase behaviour in the axilla, suggesting that the elevated cholesterol might form crystal microdomains within the lipid lamellae, allowing an increase in water flux. Since an exaggerated application of antiperspirant had no effect upon the axilla barrier properties, it is concluded that this region of skin physiologically has a reduced barrier function. Résumé La peau des aisselles est importante du point de vue cosmétique, avec des millions de consommateurs appliquant quotidiennement des produits antitranspirants/déodorants. Malgré cela, nous ne connaissons pratiquement rien de la peau des aisselles ou de l'impact que l'utilization d'antitranspirants peut avoir sur celle-ci. Afin de caractériser la stratum corneum des aisselles et de déterminer si elle constitue un type de peau spécifique, nous avons examiné la composition et la fonction de la stratum corneum, en particulier ses propriétés de barrière, comparées à d'autres zones du corps. La perte d'eau trans -épidermique (trans -epidermal water loss TEWL) et la cornéosurfamétrie (CSM) ont révélé une fonction barrière réduite dans les aisselles. L'analyse HPTLC des lipides de la stratum corneum a démontré la présence de niveaux statistiquement plus élevés d'acides gras, de céramides et en particulier de cholestérol dans la peau axillaire. Ni les céramides ni le cholestérol n'ont semblé changer en fonction de la profondeur, indiquant qu'ils sont probablement originaires du stratum corneum. Cependant, au moins une partie des acides gras avaient une origine sébacée. Notre hypothèse est qu'une réduction de la fonction de barrière de la stratum corneum axillaire est peut-être due à des changements dans le rapport crucial céramides : cholestérol. Pour examiner cette hypothèse, nous avons utilisé la spectroscopie infrarouge par transformée de Fourier ATR[l'attenuated total reflectance Fourier-transformed infrared spectroscopy (ATR,FTIR)] combinée à l'échantillonnage au cyanoacrylate. Ces résultats ont dévoilé un comportement de phase de lamelles lipidiques plus ordonnée dans les aisselles, suggérant que le cholestérol puisse former des microdomaines cristallins à l'intérieur des lamelles lipidiques, ce qui permettrait ainsi une augmentation du flux d'eau. Puisque l'application exagérée d'anti-transpirant n'a pas eu d'effet sur les propriétés de barrière des aisselles, nous concluons que cette région de la peau a une fonction de barrière physiologique réduite. [source]

Ultrastructure of the tentacle nerve plexus and putative neural pathways in sea anemones

INVERTEBRATE BIOLOGY, Issue 3 2002
Jane A. Westfall
Abstract. Neurons of sea anemone tentacles receive stimuli via sensory cells and process and transmit information via a plexus of nerve fibers. The nerve plexus is best revealed by scanning electron microscopy of epidermal peels of the tentacles. The nerve plexus lies above the epidermal muscular layer where it appears as numerous parallel longitudinal and short interconnected nerve fibers in Calliactis parasitica. Bipolar and multipolar neurons are present and neurites form interneuronal and neuromuscular synaptic contacts. Transmission electron microscopy of cross sections of tentacles of small animals, both C. parasitica and Aiptasia pallida, reveals bundles of 50,100 nerve fibers lying above groups of longitudinal muscle fibers separated by intrusions of mesoglea. Smaller groups of 10,50 slender nerve fibers are oriented at right angles to the circular muscle formed by the bases of the digestive cells. The unmyelinated nerve fibers lack any glial wrapping, although some bundles of epidermal fibers are partially enveloped by cytoplasmic extensions of the muscle cells; small gastrodermal nerve bundles lie between digestive epithelial cells above their basal myonemes. A hypothetical model for sensory input and motor output in the epidermal and gastrodermal nerve plexuses of sea anemones is proposed. [source]

Ultrastructural and histochemical study on gills and skin of the Senegal sole, Solea senegalensis

JOURNAL OF APPLIED ICHTHYOLOGY, Issue 6 2004
J. M. Arellano
Summary This study was undertaken to identify the normal ultrastructural features of gills and skin of the Senegal sole, Solea senegalensis, for a comparative measure to morphological alterations caused by environmental stressors such as reduced water quality and diseases. In the Senegal sole skin, four morphologically distinct layers were identified: cuticle, epidermis, dermis and hypodermis. The epidermis was composed of stratified epithelium containing three cellular layers: the outermost or mucosa layer, the middle or fusiform layer and the stratum germinativum or the basal layer. In the mucosa, two mucous cell types were differentiated: type A cells containing several round vesicles of different electron density and type B cells containing mucosomes of uniform electron density. Senegal sole have five pairs of gill arches, each containing two rows of well-developed and compactly organized primary filaments and secondary lamellae. Fingerprint-like microridges were observed on the surface of epithelial cells. The branchial lamellae epithelium consisted of different cell types: pavement, mucous and chloride. Between the chloride cells and the larger pavement cells, accessory cells were observed. Complexes of tight junctions and desmosomes were frequently observed between adjacent chloride and epithelial cells. Neutral mucosubstances and/or glycoconjugates were observed in the epidermis, dermis and hypodermis of S. senegalensis skin. Proteins rich in different amino acids, such as arginine and cysteine, reacted negatively or weakly positive in the epidermis, dermis and hypodermis. In gills, some mucous cells responded weakly positive to periodic acid-Schiff (PAS) reaction but were strongly stained with Alcian Blue at pH 0.5, 1 and 2.5. When Alcian Blue pH 2.5,PAS reaction was performed, most mucous cells were stained blue (carboxylated mucins) and some mucocytes stained purple, indicating a combination of neutral and acid mucins. Proteins rich in cysteine-bound sulphydryl (-SH-) and cystine disulphide (-S-S-) groups were strongly detected in branchial and epidermal mucous cells, whereas lysine, tyrosine and arginine containing proteins showed very weak staining in both epidermal and branchial mucous cells. Protein reactions were strongly positive in the pillar cells, except for those rich in tryptophan, whereas the branchial cartilaginous tissue did not show an important reaction. The performed lipid reactions were negative in goblet and chloride cells. It is concluded from this study that ultrastructural and cytohistochemical features of the Senegal sole skin and gills may serve as control structures in both natural and aquaculture systems to monitor or detect environmental stress responses at the histological level. [source]

Serological detection and immunogold localization of cross-reactive antigens shared by Camellia sinensis and Exobasidium vexans

JOURNAL OF APPLIED MICROBIOLOGY, Issue 5 2007
B.N. Chakraborty
Abstract Aims:, Pathogenicity of Exobasidium vexans, causal agent of blister blight of tea, was studied in 30 commercially cultivated tea varieties by analysing the antigenic patterns of host and pathogen using immunological techniques. Methods and Results:, Whole plant inoculation of tea varieties with E. vexans showed that T-78 and T-17/1/54 were most susceptible and most resistant respectively. Antigen preparations from tea varieties, pathogen, nonpathogen (Fusarium oxysporum) and of nonhosts (Glycine max, Leucaena leucocephala and Oryza sativa) were compared by indirect enzyme-linked immunosorbent assay and dot-immunobinding assay using polyclonal antibodies raised against the pathogen, nonpathogen, susceptible and resistant tea varieties. Cross-reactive antigens (CRA) were found among susceptible varieties and E. vexans isolates but not in resistant varieties, nonhosts or nonpathogen. Indirect staining of antibodies using fluorescein isothiocyanate indicated CRA were concentrated mainly around epidermal and mesophyll cells in compatible host (T-78). This was substantiated by ultrastructural studies using gold-labelled antibodies through transmission electron microscopy which showed specific localization in the chloroplasts and host cytoplasm. Conclusion:, Pathogenicity of E. vexans to different tea varieties is therefore related to the level of antigenic similarity between host and pathogen. Significance and Impact of the Study:, Immunological methods proved to be valuable in screening commercially cultivated tea varieties against E. vexans. [source]

Molecular and histological responses in rat skin exposed to m -xylene,

JOURNAL OF BIOCHEMICAL AND MOLECULAR TOXICOLOGY, Issue 2 2003
Palur G. Gunasekar
Abstract Solvents, surfactants, cutting fluids, hydrocarbons, and oils cause skin irritation by incompletely understood mechanisms. This study examined histological and molecular changes in rodent skin caused by brief topical exposures to m -xylene. At 0, 1, 2, 4, and 6 h after 1-h exposure, skin samples were removed and analyzed for histopathological changes and interleukin-1, (IL-1,) and inducible nitric oxide synthase (iNOS) protein levels. Histopathological changes (epidermal,dermal separation and granulocyte infiltration) and increases in IL-1, and iNOS protein expression occurred during our observation period. IL-1, levels increased by 80% immediately after exposure and iNOS levels increased about 60% 4 hours after exposure. Our study demonstrates that dermal exposure to m -xylene promotes IL-1, and iNOS production in skin and these proteins may serve as early indicators of skin irritation. © 2003 Wiley Periodicals, Inc. J Biochem Mol Toxicol 17:92,94, 2003; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jbt.10065 [source]

A biodegradable copolymer for the slow release of growth hormone expedites scarring in diabetic rats

JOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 2 2007
Francisco García-Esteo
Abstract In many diseases wound healing is impaired. This study was designed to establish whether the healing process in diabetes could be improved using a site-specific polymer delivery system containing hGH. The system was first optimized in in vitro experiments performed on cultured fibroblasts taken from healthy and diabetic rats and then tested in an incisional wound model created in the diabetic Wistar rat. In the in vitro experiments using cultured fibroblasts, cell viability, growth, and proliferation were determined, along with polymer degradation, hormone release rates and the expression of TGF,1 in the culture medium. For the in vivo experiments, polymer discs with/without GH were inserted through 3 cm incisions made on the backs of the animals. Wound specimens were obtained 7 and 30 days after surgery to evaluate inflammatory/apoptotic cells, metalloprotease expression and neoangiogenesis using microscopy and immunohistochemical techniques. The local administration of GH using a polymer delivery system did not affect the normal wound healing process. Conversely, when used in diabetic animals, epidermal and dermal repair was expedited. Our findings indicate that GH induces cell proliferation, enhances CD4+ infiltration; increases extracellular matrix protein deposition; stimulates angiogenesis; and diminishes apoptosis at the diabetic wound site. These effects give rise to a comparable wound healing process to that observed in healthy animals. © 2006 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2006 [source]

A double-blind controlled study of a nonhydroquinone bleaching cream in the treatment of melasma

JOURNAL OF COSMETIC DERMATOLOGY, Issue 4 2005
Jean Luc Levy
Summary Background, Melasma is an acquired hypermelanosis predominantly affecting the face of women. It is often recalcitrant to treatment with hypopigmenting agents. Objective, To assess the efficacy of a nonhydroquinone cream (Amelan M®) vs. another (Mela-D®) as treatment for melasma. Methods, Twenty-two French women with bilateral epidermal and/or mixed melasma were enrolled in a split-faced prospective trial lasting 4 months during summer season weeks. Twelve patients applied once-daily Amelan M® to one side of the face with sun-protective factor 60 UVA sunscreen each morning and Mela-D® once-daily to the other side of the face. Pigmentation was measured objectively using a mexameter and the melasma area and severity index (MASI) were measured subjectively. Results, The mean decrease of pigmentation was statistically significant on the MASI with both cream and only with Amelan M with the mexameter. Some adverse side effects were observed. Conclusions, Amelan M® is really more effective than Mela D® cream on melasma. Even though some side effects were observed patients preferred the Amelan M-treated side. [source]

Herpetic Cytopathic Features Confined to Folliculosebaceous Units: What does it Mean?

JOURNAL OF CUTANEOUS PATHOLOGY, Issue 1 2005
N Walsh
The histopathological changes of herpes simplex, zoster and varicella are considered to be indistinguishable from one another. The clinical setting with adjunctive studies generally clarifies the diagnosis. Vesicular lesions in all 3 conditions can involve epidermal and adnexal epithelium with characteristic cytopathic features. We describe 3 patients with non-vesicular eruptions on the head and neck whose biopsies revealed exclusive folliculosebaceous involvement by herpes. All three patients developed typical herpes zoster within days of the biopsy. There is compelling scientific evidence in the literature indicating that, in herpes zoster, the virus is transported from dorsal root or trigeminal ganglia via myelinated nerves to the skin. These terminate at the isthmus of hair follicles and primary infection of follicular and sebaceous epithelium occurs. Secondary spread of infection to the epidermis follows. In contrast, data pertaining to recurrent herpes simplex indicates that axonal transport of the virus from sensory ganglia to the skin is directed primarily to the epidermis, via terminal non-myelinated nerve twigs. The clinical evolution of our 3 cases and scientific data in the literature indicate that exclusive folliculosebaceous involvement by herpes, in the setting of a non-vesicular eruption, represents early herpes zoster. [source]