Physical Irritation Research Articles

Research Progress of Far Field Light Scattering Spectra of Single Gold Nanorods

The far-field optical technology of single gold-nanorod(AuNR) has attracted a great deal of attention in recent years. Because of special local surface plasmon resonance (LSPR) property, AuNR particles have high conductivity of optical signals localized on the surface from physical or chemical irritants. The mechanism, application, progress and novel optical characteristics of AuNRs in optical detection and spectroscopy method are being reviewed in our work. The paper describes an overall introduction as follows: (1)various related technologies on AuNR scattered spectrum, including dark-field technology, homodyne and heterodyne technology, photonic crystal technology, spatial modulation, polarization modulation technology, etc; (2) the properties of AuNR scattered spectrum, including spectral line-shape functions, effects of line-width and substrate, comparison of theoretical and experimental spectrums, etc; (3) the development of related spectrum technologies in recent years. The paper focuses on the method of far-field optical scattering based on LSPR and mainly discusses the linear method based on AuNRs, such as direct and indirect scattering detection method. We also put emphasis upon studying the importance of medium environment (for example, substrate, the molecules combined on surface and other nanomaterials) and the influence on scattered spectrum and the extinction rate. Of particular note is the quantitative method and correlation studies of AuNR’s surface and morphology, and its character is that most of the methods are compared with theoretical model and experiments in terms of accuracy. The combination of the experiments and theoretical tools can be used to explain the optical properties of single gold-nanorod particle in detail.

CONSIDER - Core Outcome Set in IAD Research: study protocol for establishing a core set of outcomes and measurements in incontinence-associated dermatitis research.

This study protocol describes the methodology for the development of a core set of outcomes and a core set of measurements for incontinence-associated dermatitis. Incontinence is a widespread disorder with an important impact on quality of life. One of the most common complications is incontinence-associated dermatitis, resulting from chemical and physical irritation of the skin barrier, triggering inflammation and skin damage. Managing incontinence-associated dermatitis is an important challenge for nurses. Several interventions have been assessed in clinical trials, but heterogeneity in study outcomes complicates the comparability and standardization. To overcome this challenge, the development of a core outcome set, a minimum set of outcomes and measurements to be assessed in clinical research, is needed. A project team, International Steering Committee and panelists will be involved to guide the development of the core outcome set. The framework of the Harmonizing Outcomes Measures for Eczema roadmap endorsed by Cochrane Skin Group Core Outcomes Set Initiative, is used to inform the project design. A systematic literature review, interviews to integrate the patients' perspective and a consensus study with healthcare researchers and providers using the Delphi procedure will be performed. The project was approved by the Ethics review Committee (April 2016). This is the first project that will identify a core outcome set of outcomes and measurements for incontinence-associated dermatitis research. A core outcome set will reduce possible reporting bias, allow results comparisons and statistical pooling across trials and strengthen evidence-based practice and decision-making. This project has been registered in the Core Outcome Measures in Effectiveness Trials (COMET) database and is part of the Cochrane Skin Group Core Outcomes Set Initiative (CSG-COUSIN).

Patterns and severity of menopausal symptoms among Jordanian women

ABSTRACTWe examined 359 women aged 45–65 years who visited Jordan University Hospital between February and November 2014. The menopausal symptoms were assessed using a validated Arabic version of the menopause rating scale. The mean age at menopause was 49.4 years. Women aged 50–55 years more frequently exhibited hot flushes and vaginal dryness. Although premenopausal women were 1.5 times more likely to experience irritability, perimenopausal women were more likely to experience hot flushes, physical and mental irritability, sexual problems, vaginal dryness, and joint and muscular discomfort. Hence, health care providers should focus on women at all stages of life.

Transcriptome Sequencing of Chemically Induced Aquilaria sinensis to Identify Genes Related to Agarwood Formation.

BackgroundAgarwood is a traditional Chinese medicine used as a clinical sedative, carminative, and antiemetic drug. Agarwood is formed in Aquilaria sinensis when A. sinensis trees are threatened by external physical, chemical injury or endophytic fungal irritation. However, the mechanism of agarwood formation via chemical induction remains unclear. In this study, we characterized the transcriptome of different parts of a chemically induced A. sinensis trunk sample with agarwood. The Illumina sequencing platform was used to identify the genes involved in agarwood formation.Methodology/Principal FindingsA five-year-old Aquilaria sinensis treated by formic acid was selected. The white wood part (B1 sample), the transition part between agarwood and white wood (W2 sample), the agarwood part (J3 sample), and the rotten wood part (F5 sample) were collected for transcriptome sequencing. Accordingly, 54,685,634 clean reads, which were assembled into 83,467 unigenes, were obtained with a Q20 value of 97.5%. A total of 50,565 unigenes were annotated using the Nr, Nt, SWISS-PROT, KEGG, COG, and GO databases. In particular, 171,331,352 unigenes were annotated by various pathways, including the sesquiterpenoid (ko00909) and plant–pathogen interaction (ko03040) pathways. These pathways were related to sesquiterpenoid biosynthesis and defensive responses to chemical stimulation.Conclusions/SignificanceThe transcriptome data of the different parts of the chemically induced A. sinensis trunk provide a rich source of materials for discovering and identifying the genes involved in sesquiterpenoid production and in defensive responses to chemical stimulation. This study is the first to use de novo sequencing and transcriptome assembly for different parts of chemically induced A. sinensis. Results demonstrate that the sesquiterpenoid biosynthesis pathway and WRKY transcription factor play important roles in agarwood formation via chemical induction. The comparative analysis of the transcriptome data of agarwood and A. sinensis lays the foundation for elucidating the mechanism of agarwood formation via chemical induction, and thus, enables future improvements in agarwood quality while protecting endangered wild A. sinensis.

Irritant contact dermatitis is regulated by antioxidative enzyme NAD(P)H:quinone oxidoreductase (Nqo1).

Abstract Irritant contact dermatitis (ICD) is the most common occupational skin inflammatory disorder caused by the exposure of chemical or physical irritants. Local release of inflammatory mediators such as cytokines, lipid mediators, and reactive oxygen species (ROS) are known to be important in the establishment of ICD. Antioxidative enzymes such as Nqo1 are known to be responsible for the degradation of ROS. Therefore, in the current study, we focused on the involvement of Nqo1 in ICD. In the mouse ICD model using croton oil, we found that Nqo1-deficient mice exhibited increased ear swelling and augmented expression of proinflammatory cytokines such as IL-6 and IL-17A in the skin. As we expected, it is indicated that Nqo1 has protective role against ICD. However, it is not applicable for general inflammatory responses, because the knockout mice showed unaltered response in the allergic contact dermatitis mouse model. Interestingly, we found that severe reduction of Vγ5Vδ1+ dendritic epidermal T cells (DETCs) in the skin of Nqo1 deficient mice, which are reported to inhibit ICD. Transfer of DETCs into RAG−/− mice revealed impaired survival of Nqo1-deficient DETCs. In addition, Nqo1-deficient DETCs were sensitive to oxidative stress-induced cell death, and anti-oxidant NAC treatment rescued decreased DETCs in Nqo1-deficient mice. These results suggest that Nqo1 regulates ICD through DETC development and/or homeostasis, possibly by antioxidative function.

A decade of research on Incontinence-Associated Dermatitis (IAD): Evidence, knowledge gaps and next steps

BackgroundIncontinence-Associated Dermatitis (IAD) is one of the clinical manifestations of Moisture- Associated Skin Damage (MASD). IAD is a common problem in aged patients with fecal and/or urinary incontinence. AimUpdate about IAD terminology, etiology, epidemiology, observation, prevention, and treatment. MethodsIntegrative review. ResultsThe lack of an ICD-10 code and an internationally validated and standardized method for IAD data collection contribute to a variation in epidemiological data. Frequent episodes of incontinence (especially fecal), occlusive containment products, poor skin condition, reduced mobility, diminished cognitive awareness, inability to perform personal hygiene, pain, pyrexia, certain medications (antibiotics, immunosuppressants), poor nutritional status, and critical illness are associated with IAD. Correctly diagnosing IAD and distinguish it from pressure ulcers is difficult. Even though the clinical presentation of partial thickness pressure ulcers and IAD is similar, the underlying etiologic factors differ. However, incontinence and IAD were found to be risk factors for pressure ulcer development. IAD management should essentially focus on skin cleansing to remove dirt, debris and microorganisms; skin moisturization to repair or augment the skin's barrier; and the application of a skin barrier product to prevent skin breakdown by providing an impermeable or semi-permeable barrier on the skin. The body of evidence is still limited, but growing since the last decade. ConclusionIncontinence causes disruptions of the skin barrier function and leads to superficial skin damage. Macerated skin and superficial skin changes due to incontinence are associated with pressure ulcer development. Skin maceration, chemical irritation, and physical irritation should be targeted to effectively prevent and treat IAD.

Sleep and Behavioral Problems in Children with Autism Spectrum Disorder.

Children with autism spectrum disorder (ASD) are at high risk for sleep disturbance and behavioral dysregulation. However, the relationships between these difficulties are not fully understood. The current study examined the relationships between specific types of sleep and behavioral problems among 81 children with ASD. Sleep problems were significantly associated with physical aggression, irritability, inattention, and hyperactivity. In multivariate analyses, distinct sets of sleep problems accounted for between 22 and 32% of the variance in behavior problems across models. These results indicate that sleep disturbance is associated with behavioral dysregulation among children with ASD. Of note, night awakenings had the most consistently strong association with daytime behavior problems, even after controlling for the effects of age and sex.

Evaluasi Uji Iritasi Dan Uji Sifat Fisik Sediaan Emulgel Minyak Atsiri Bunga Cengkeh (Syzigium Aromaticum)

Inflammation is the human body's response to a tissue damage as indicated by an increase of vascular permeability. One of the natural substances that can be used to treat inflammation is clove oil (MABC). This study aimed to evaluate the physical properties and MABC emulgel irritation test ( S yzygium aromaticum ). This experimental study was initiated by the emulgel formulation with three kinds of formula, with each formula contains MABC levels of 10% (F1), 12.5% (F2), and 15% (F3). The physical tests for emulgel was done including pH, spreading test and adhesive test and irritation test in guinea pigs using Draize test method. Statistical analysis by t test with a level of confidence 95% was conducted to detect a significant differences between the treatment groups. The emulgel formula with a concentration of 10%, 12.5% and 15% were physically stable, moreover the dispersive power test for each formula showed normal distribution data (p>0.05) and homogeneous (p>0.05), ANOVA test results obtained significantly different results in each formula (p<0.05), whereas the stickiness test results for each formula more than 30 minutes with pH 6. The results of irritation test by using Draize test method, indicated that each formula did not show any irritant effect.

Victimisation, limbic system irritability and sociomoral reasoning in male sex offenders

ABSTRACTThis study used Narvaez's (2010) Triune Ethics Theory to examine the victim-to-perpetrator hypothesis. The study surveyed 68 sex offenders from the New York State Sex Offender Registry to investigate correlates of physical and sexual abuse, and to analyse associations between victimisation, limbic system irritability, and sociomoral reasoning. Surveys were mailed to sex offenders and analysed using correlation and multivariate analysis of variance. Post hoc analysis of Sexual and Physical Abuse Questionnaire and Limbic System Check List-33 scores showed a significant relationship between physical and sexual abuse and limbic system irritability. Using the Sociomoral Reflection Measure-Short Form, sex offenders were found on average to reason at the Transitional stage 2(3). Predicting self-report abuse history resulted in a 35% correct classification rate based on within-group correlations between predictors and discriminate functions as well as standardised weights.

Cell Death and Proliferation after Treatment and Reinfection of Clonorchis sinensis in the Sprague-Dawley Rat Bile Duct

The structural change and distribution of mitochondrial enzyme (ATPase, cytochrome-c-oxidase), cell proliferation (proliferating cell nuclear antigen, PCNA), cell death (caspase-3) and cell growth factor (fibroblast growth factor 8, FGF-8) in the Sprague-Dawley rat bile duct during Clonorchis sinensis infection was investigated. Experimental groups were divided into C. sinensis infection, superinfection and reinfection of C. sinensis after `praziquantel` treatment group. As a result, C. sinensis infected rat bile ducts showed the features of chronic clonorchiasis, i.e., connective tissue thickening, ductal fibrosis and epithelial tissue dilatation. PCNA for cell proliferation increased in the infection group, and decreased after praziquantel treatment. Caspase-3 was distributed in reinfection group only. FGF-8 was distributed in the rat bile duct after praziquantel treatment but not distributed in infection and reinfection group. Overall, C. sinensis infection causes physical and chemical irritations and then brings on the abnormalities of intracellular energy metabolism and cellular growth factors, which hinders bile duct tissue from functioning properly, and resultingly, fibrosis occurs and epithelial cells dilated abnormally. More intense infection makes tissue fibrosis chronical and activates apoptosis factors.

Nasal citology: a new diagnostic approach in rhinology

Nasal cytology is a diagnostic method in the field rinology used to detect changes into the endonasal epithelium exposed to physical and/or chemical irritation, acute or chronic inflammation due to different angents (viral, bacterial, fungal or parasitic). Nasal cytology was born in 1889 thanks to H. Gollash who found many eosinophilic cells into the nasal secretions of a patient affected by asthma, and thought of them as important elements in the pathogenesis of this disease. The main evolution of nasal cytology occurred in 1927, thanks to the report done by C. Eyermann, who noted the presence of eosinophilic cells into the nasal secretions of allergic patients. Since then great importance is given to the recognition of specific cell types into the pathogenesis of different nasal diseases. Nasal cytology is nowadays more frequently used in the study of allergic, vasomotor, inflammatory and infectious rhinitis. It is a simple, non-invasive and repeatable examination which is useful in the follow-up and monitoring of the real effectiveness of medical and surgical treatments. Patients affected by allergic rhinitis (AR) develop an endonasal immediate response, so-called “early phase”, followed by a “late phase” response to the allergenic agent. From the microscopic point of view, both these responses are always characterized by an infiltration of the mucosa by immunoflogistic cells (eosinophils, mast cells, neutrophils and lymphocytes) that, following the release of several chemical mediators, are the main cause of the symptoms that characterize the IgE-mediated disease. In AR the main cells type released during the immunological response will change depending on whether the patient is examined during or outside the pollinic season. In the first condition, the patient will present all the clinical signs and his nasal cytology examination will be characterized by neutrophils, lymphocytes, eosinophils and mast cells largely degranulated. Nasal cytology allows us to differentiate between various forms of rhinitis which are resumed as: non-allergic rhinitis with neutrophil (Narne); non-allergic rhinitis with eosinophilia (NARES); non-allergic rhinitis with mast cells (NARMA); non-allergic eosino.

Genital allergy and irritant reactions in atopics and non‐atopics

Genital allergy or irritant reaction should be considered as a possible diagnosis in all patients with genital soreness or irritation for which no infection or dermatosis can be identified and in whom clinical manifestations remain unchanged or worsen with treatment. The term denotes an acute or chronic inflammatory skin disorder caused by chemical/ physical irritants - irritant contact dermatitis (ICD) or allergens - allergic contact dermatitis (ACD). Despite their different mechanisms, they can be difficult to distinguish from one another at the clinical, histology, and even molecular levels. Clinical manifestations may be associated with irritants. They have the potential to disrupt membranes or interfere with metabolic processes in the epidermis or dermis. Chemical ICD (CICD) is either acute or chronic, which is usually associated with strong and weak irritants, respectively. Highly alkaline soaps, detergents, and cleaning products all have the common efffect of directly affecting the barrier properties of the epidermis. These effects include removing fat emulsion, inflicting cellular damage on the epithelium, and increasing the transepidermal water loss (TEWL) by damaging the horny layer water-binding mechanisms and damaging the DNA, which causes the layer to thin. The cosmetic products responsible for irritation are mainly liquid foam hygiene products. used too frequently and/or too long and/or badly rinsed. In woman, depilatory products can also be to blame. Some patients complain of irritation due to the perfume of scented toilet papers. Physical ICD (PICD) is a less researched form of ICD due to its various mechanisms of action and a lack of tests for its diagnosis. A complete patient history combined with negative allergic patch testing is usually necessary to reach a correct diagnosis. The simplest form of PICD results from prolonged rubbing, although the diversity of implicated irritants is far wider. Examples include paper friction and scratchy clothing. The anogenital region is often the site of ICD: in men, promoted by sweat and rubbing and in woman, by aggressive hygiene. ICD affects very young and very old patients more severely. The greatest single risk for ICD is a history of atopic dermatitis.

Dont Sweat It: Treating Hyperhidrosis with Microwaves

When considering the medical applications of microwave technology, ablating sweat glands is probably not what initially comes to mind. However, hyperhidrosis (the medical term for excessive sweating) is a condition estimated to affect up to 3% of the population [1]. There is also evidence that an additional 17% of the population have symptoms that are similar to those diagnosed with the condition [2]. Imagine a scenario where someone is inactive, sitting at his or her desk in a cool, dry environment, yet is sweating from the hands, feet, or underarms as if exercising. This is the type of situation a hyperhidrosis sufferer might face on a daily basis. Beyond physical irritation, there is significant evidence that the condition can result in occupational, emotional, psychological, social, and physical impairment leading to increased amounts of depression and other negative effects [1]. The treatment of sweat is also common in the general population in addition to severe sufferers, as evident from the large amounts of money spent on deodorants and antiperspirants (e.g., US$2.9 billion in the United States in 2012) [3]. In this article, an overview of the current treatment options for axillary (underarm) hyperhidrosis, potential energy-based approaches, and the microwave technique developed for treating this potentially debilitating condition is presented. The challenges of commercializing such a medical microwave device are also discussed.

Extracellularly Extruded Syntaxin-4 Is a Potent Cornification Regulator of Epidermal Keratinocytes.

In the skin epidermis, keratinocytes undergo anchorage-dependent cornification, which gives rise to stratified multilayers, each with a distinct differentiation feature. The active formation of the cornified cell envelope (CCE), an important element in the skin barrier, occurs in keratinocytes of the upper epidermal layers and impacts their terminal differentiation. In the present study, we identified the extracellularly extruded syntaxin-4 as a potent differentiation regulator of epidermal keratinocytes. We found that differentiation stimuli led to the acceleration of syntaxin-4 exposure at the keratinocyte cell surface and that the artificial control of extracellular syntaxin-4, either by the forced expression of several syntaxin-4 mutants with structural alterations at the putative functional core site (AIEPQK), or by using antagonistic circular peptides containing this core sequence, dramatically influenced the CCE formation, with spatial misexpression of TGase1 and involucrin. We also found that the topical application of a peptide that exerted the most prominent antagonistic activity for syntaxin-4, named ST4n1, evidently prevented the formation of the hyperplastic and hyperkeratotic epidermis generated by physical irritation in HR-1 mice skin. Collectively, these results demonstrate that extracellularly extruded syntaxin-4 is a potent regulator of CCE differentiation, and that ST4n1 has potential as a clinically applicable reagent for keratotic skin lesions.

Induction of cancer-related microRNA expression profiling using excretory-secretory products of Clonorchis sinensis.

Clonorchis sinensis is a carcinogenic human liver fluke by which chronic infection is strongly associated with the development of cholangiocarcinoma. Although this cholangiocarcinoma is caused by both physical and chemical irritation from direct contact with adult worms and their excretory-secretory products (ESPs), the precise molecular events of the host-pathogen interactions remain to be elucidated. To better understand the effect of C. sinensis infection on cholangiocarcinogenesis, we profiled the kinetics of changes in cancer-related microRNAs (miRNAs) in human cholangiocarcinoma cells (HuCCT1) treated with C. sinensis ESPs for different periods. Using miRNA microarray chips containing 135 cancer-related miRNAs, we identified 16 miRNAs showing differentially altered expression following ESP exposure. Of these miRNAs, 13 were upregulated and 3 were downregulated in a time-dependent manner compared with untreated controls. Functional clustering of these dysregulated miRNAs revealed involvement in cell proliferation, inflammation, oncogene activation/suppression, migration/invasion/metastasis, and DNA methylation. In particular, decreased expression of let-7i, a tumor suppressor miRNA, was found to be associated with the ESP-induced upregulation of TLR4 mRNA and protein, which contribute to host immune responses against liver fluke infection. Further real-time quantitative PCR analysis using ESP-treated normal cholangiocytes (H69) revealed that the expressions of nine miRNAs (miR-16-2, miR-93, miR-95, miR-153, miR-195, miR-199-3P, let7a, let7i, and miR-124a) were similarly regulated, indicating that the cell proliferation and inhibition of tumor suppression mediated by these miRNAs is common to both cancerous and non-cancerous cells. These findings constitute further our understanding of the multiple cholangiocarcinogenic pathways triggered by liver fluke infection.

Phenylboronic acid modified mucoadhesive nanoparticle drug carriers facilitate weekly treatment of experimentallyinduced dry eye syndrome

Topical formulations, commonly applied for treatment of anterior eye diseases, require frequent administration due to rapid clearance from the ocular surface, typically through the lacrimal drainage system or through over-spillage onto the lids. We report on a mucoadhesive nanoparticle drug delivery system that may be used to prolong the precorneal residence time of encapsulated drugs. The nanoparticles were formed from self-assembly of block copolymers composed of poly(d, l-lactide) and Dextran. The enhanced mucoadhesion properties were achieved by surface functionalizing the nanoparticles with phenylboronic acid. The nanoparticles encapsulated up to 12 wt.% of Cyclosporine A (CycA) and sustained the release for up to five days at a clinically relevant dose, which led us to explore the therapeutic efficacy of the formulation with reduced administration frequency. By administering CycA-loaded nanoparticles to dry eye-induced mice once a week, inflammatory infiltrates were eliminated and the ocular surface completely recovered. The same once a week dosage of the nanoparticles also showed no signs of physical irritation or inflammatory responses in acute (1 week) and chronic (12 weeks) studies in healthy rabbit eyes. These findings indicate that the nanoparticles may significantly reduce the frequency of administration for effective treatment of anterior eye diseases without causing ocular irritation.

PENGEMBANGAN FORMULASI MASKER EKSTRAK AIR DAUN ALPUKAT (Persea americana Mill) SEBAGAI ANTIBAKTERI Staphylococcus aureus UNTUK PENGOBATAN JERAWAT

Skin problems are often on the spotlight, one of them is acne. Inflammation of acne can be triggered by the bacteria Propionibacterium acne, Staphylococcus epidermidis and Staphylococcus aureus . One of the herbs that can be used for acne treatment is the leaves of avocado ( Persea americana ). This study aims to determine the potential antibacterial activity of avocado leaf water extract on the growth of Staphylococcus aureus bacteria that causes acne and mask-making formulation preparation. Antibacterial activity test was carried out by the diffusion method and the observed inhibition zone that occured in a series of concentrations of avocado leaf water extract ( 15%, 17.5% , 35% , 50% , 75%). Antibacterial activity test was also conducted on some avocado leaf water extract mask formula (17.5% , 35% , 50% , 75%) was evaluated the physical properties of the mask and irritation test was done. The results showed that the avocado leaf water extract mask formula had antibacterial activity greater than the extract form. The physical condition evaluation and irritation test showed that the best mask formulation was concentration of 35%.

Formulation and evaluation of Cyclosporin A emulgel for ocular delivery

Emulgels have been extensively covered as a promising drug delivery system for the administration of lipophilic drugs. This work was conducted to develop an emulgel formulation for Cyclosporin A (CsA) employing polycarbophil as the gelling agent for ocular delivery. The prepared emulgels were evaluated for their physical appearance, rheological behavior, drug release, stability, precorneal clearance and irritation. Results showed that CsA emulgel formulations prepared with polycarbophil exhibited acceptable physical properties and drug release, which remained consistent after storage for 3 months. A prolonged retention time was also observed on the ocular surface with improved ocular bioavailability and no irritation. Therefore, the polycarbophil-based emulgel could be exploited as a potential hydrophobic drug carrier for topical ocular drug delivery.

Increased expression of bronchial epithelial transient receptor potential vanilloid 1 channels in patients with severe asthma

The airway epithelium is exposed to a range of physical and chemical irritants in the environment that are known to trigger asthma. Transient receptor potential (TRP) cation channels play a central role in sensory responses to noxious physical and chemical stimuli. Recent genetic evidence suggests an involvement of transient receptor potential vanilloid 1 (TRPV1), one member of the vanilloid subfamily of TRP channels, in the pathophysiology of asthma. The functional expression of TRPV1 on airway epithelium has yet to be elucidated. In this study we examined the molecular, functional, and immunohistochemical expression of TRPV1 in asthmatic and healthy airways. Bronchial biopsy specimens and bronchial brushings were obtained from healthy volunteers (n = 18), patients with mild-to-moderate asthma (n = 24), and patients with refractory asthma (n = 22). Cultured primary bronchial epithelial cells from patients with mild asthma (n = 4), nonasthmatic coughers (n = 4), and healthy subjects (n = 4) were studied to investigate the functional role of TRPV1. Quantitative immunohistochemistry revealed significantly more TRPV1 expression in asthmatic patients compared with healthy subjects, with the greatest expression in patients with refractory asthma (P = .001). PCR and Western blotting analysis confirmed gene and protein expression of TRPV1 in cultured primary bronchial epithelial cells. Patch-clamp electrophysiology directly confirmed functional TRPV1 expression in all 3 groups. In functional assays the TRPV1 agonist capsaicin induced dose-dependent IL-8 release, which could be blocked by the antagonist capsazepine. Reduction of external pH from 7.4 to 6.4 activated a capsazepine-sensitive outwardly rectifying membrane current. Functional TRPV1 channels are present in the human airway epithelium and overexpressed in the airways of patients with refractory asthma. These channels might represent a novel therapeutic target for the treatment of uncontrolled asthma.

Primary culture of human face skin melanocytes for the study of hyperpigmentation

Facial epidermal pigmentation and skin tumors can be caused by UV exposure and other physical and chemical irritations. In this report we describe the primary culture of melanocytes from human face skin. The ability to culture these melanocytes will enable their morphological and biological properties to be investigated. Skin specimens were obtained from patients who had undergone lower blepharoplasty procedures. Digestion with neutral protease and trypsin was used to obtain single cell suspensions of epidermal cells. The cells were cultured in M254 medium supplemented with human melanocyte growth solution. Cell morphology was observed using inverted microscopy. Melanocytes were positively identified using both L-DOPA staining and S-100 protein immunohistochemical staining. Immunofluorescence was used to confirm the expression of tyrosinase-related protein-1, a melanocyte-specific protein. The cellular ultrastructure of the melanocytes was observed by transmission electron microscopy. The cultured human melanocytes from face skin were multi-dendritic, and many mature melanosomes were observed. Therefore, using a specific culture medium, melanocytes from face skin can be successfully cultured and made available for further investigations.