Skin Surface Area Research Articles

Antibiotic route and duration of therapy for cellulitis: data extracted from a multi-center clinical trial

Introduction: Although cellulitis is a relatively common condition, there is uncertainty about the benefit of intravenous (IV) over oral (PO) antibiotic therapy, and the appropriate duration of treatment.Methods: Data extracted from a clinical trial (NCT01876628) of antibiotic therapy for cellulitis were used to assess the association between the route of administration and duration of treatment, and clinical outcome.Results: Of 323 patients with antibiotic data, 114 received some IV therapy. IV antibiotic therapy was preferred in those who had received antibiotics prior to trial entry (P < 0.001). Patients characterised as having more severe cellulitis (C-reactive protein > 100 mg/L, affected skin surface area > 5% or systemic inflammatory response syndrome score ≥ 1) were more likely to have had IV therapy. Those given only PO therapy were more likely to have improved at day 5 compared with those given at least a single dose of IV therapy (P = 0.015), and were as likely to be back to their normal activities at day 10 (P = 0.90), and day 30 (P = 0.86). There was no association between initial severity and the duration of antibiotic therapy given within the trial. There was no association between duration of antibiotic therapy and outcome as measured at day 10 and day 30.Conclusions: This study provides evidence that recovery is not associated with the route of antibiotic administration for patients with cellulitis of similar severity, or that a course length of > 5 days results in any additional benefit.

Topical delivery of siRNA into skin using ionic liquids

Skin diseases such as lupus, cancer, psoriasis, and hyperhidrosis can potentially be treated effectively by suppressing allele-specific genes using small interfering RNA (siRNA). Injections of siRNA into skin, though effective, are painful and cover small surface areas and thus are not suitable as a long-term treatment option. Topical delivery of siRNA is an attractive alternative option to mediate RNA interference (RNAi). However, the barrier function of the epidermis impedes effective permeation of siRNA into the skin. Herein, we describe topical delivery of siRNA using ionic liquids (ILs) capable of complexing with siRNA non-covalently and delivering it effectively. Using complementary and synergistic strategies of ionic liquids, we report delivery of effective doses of siRNA into skin. The first strategy involved the use of hydrophobic cations to robe the siRNA and the second strategy involved the use of choline-geranic acid ionic liquid (CAGE) to enhance its dermal penetration. In vitro studies in porcine skin confirmed the synergistic effect of these strategies in enhancing epidermal and dermal penetration. In vivo application of siRNA formulation to SKH-1E hairless mice significantly suppressed GAPDH expression with no clinical evidence of toxicity. This is a simple, personalized, and scalable platform for effective topical delivery of siRNA for treating genetic skin diseases.

Correlation of neurological level and sweating level of injury in persons with spinal cord injury

Objective: Thermoregulatory dysfunction after spinal cord injury (SCI) impairs quality of life and predisposes persons to life-threatening sequela of heat-related illness (HRI) in conditions of high ambient temperature. SCI clinicians currently have no objective way to predict which persons are at greatest risk of HRI. Evaporative cooling via sweating is the body’s most efficient mechanism of heat dissipation. The relationship between the neurological level of injury (NLOI) and the degree of sudomotor dysfunction is not well defined. This study examines the relationship between the NLOI and sweating level of injury (SwLOI). This information can assist SCI clinicians in identifying individuals with SCI who have most impaired sudomotor function and thus highest risk of HRI. Design: Observational. Setting: Human physiology laboratory. Participants: 10 persons with tetraplegia (TP), 14 with paraplegia (PP) and 10 able-bodied (AB). Intervention: Passive heat stress (1°C rise in core temperature) with sweat responses (SR) quantified with the starch iodine test. Outcome measures: The most caudal dermatomal level in which sweating was visualized was recorded as the SwLOI, which was compared to the NLOI. Minimum, maximum and median differences between NLOI and SwLOI were calculated. Results: Persons with tetraplegia demonstrated no SR. Persons with paraplegia demonstrated SR at a median of 1 level below NLOI. Able-bodied controls demonstrated sweating on all skin surface areas. Conclusions: Persons with motor complete tetraplegia lack evaporative cooling capacity through SR during passive heat stress predisposing them to HRI. Meanwhile, persons with paraplegia sweat on average 1 dermatomal level below their NLOI.

Sobol sensitivity approach for the appraisal of geomedical health risks associated with oral intake and dermal pathways of groundwater fluoride in a semi-arid region of south India.

The main objective of the present investigation is appraisal of human health hazard based on the intake and dermal contact of fluoride enriched potable groundwater used for rural water supply in a semi-arid region (Shanmuganadhi River basin) of south India. A variance decomposition based Sobol sensitivity method was used to assess the relative contribution as well as interaction of input variables for both oral and dermal models. Three different scores were evaluated: FOE (first order effect), SOE (second order effect) and TE (total effect) for different age groups of population including gender (kids, women and men). The spatio-temporal mapping indicates that about 26% of water supply wells exceeded the recommended limit (WHO) of fluoride (>1.5mgl-1) for safe intake. These wells spread over 104.03km2 area consisting 16 villages in the basin. To assess the human health risk related to fluoride enrichment in potable water, hazard index (HI) was calculated as per USEPA guidelines. The non-carcinogenic risk based on oral intake ranges from 0 to 1.81, from 0 to 1.59 and from 0 to 1.29 for kids, women and men respectively. Nearly 30%, 21% and 12% of well samples exceeded the upper permissible limit (HI>1) for kids, women and men respectively. The Sobol sensitivity analysis reveals that, Cw (concentration of F- in water) and IR (intake rate) combination plays a vital role in the HQ oral model for the appraisal of health hazard in kids. However, these two parameters have negligible effect on health hazard for adult population (men and women). Therefore, lower age group people especially kids have significant ill effect due to the consumption of fluoride enriched potable water. The model output suggests that body weight (BW) has least effect on health hazard in the lower age group of population. Interestingly 'exposure frequency (EF)' and 'body weight (BW)' have not much effect on health hazards related to dermal contact, and 'skin surface area (SA)' has minimal effect for men (TE=0.92) and women (TE=1.26). However, 'SA' has vital effect for kids (TE=17.39). Because of this, older age group people have more dermal risk than the younger people. Therefore, the HQ dermal results indicate that 49%, 64% and 67% of samples possess non-carcinogenic risk to the kids, women and men respectively.

Scrotal thermography to assess the seminal quality in Nelore and Girolando bulls

Bull selection by andrological examination aims to estimate the reproductive capacity of the male. Bulls of zebu origin adapt better to high temperatures than bulls of taurine origin, which may influence scrotum temperature and seminal quality due to the imbalance of testicular thermoregulation. The objective of this study was to investigate the relationship between bioclimatic variables, the temperature of body and scrotal areas, assessed with infrared thermography, and the quality of fresh and post-thawed semen in zebu Nelore bulls (Bos taurus indicus) and Girolando bulls (Bos taurus taurus x Bos taurus indicus). Bulls were kept in pickets with access to water, mineral mix and a diet supplemented with concentrate. Infrared thermographs of the scrotum, orbital globe and muzzle were performed twice a week with a Flir E40 thermal imager. For scrotal thermograms, we analyzed the temperatures of the spermatic cord, proximal and distal portion of the testes and tail of the epididymis using the Flir Tools software. Samples were collected using an artificial vagina and the ejaculates were processed and frozen in liquid nitrogen until further analyses. Data were analyzed with the Tukey test or the Kruskall-Wallis test, depending on their normal distribution. Our results showed differences (p &lt; 0.05) between the two breeds regarding the temperature in the ocular globe, spermatic cord and proximal portion of the scrotum. Nelore bulls presented lower temperature in the body and in certain regions of the scrotum compared to Girolando bulls. Seminal characteristics varied between breeds, with the Nelore breed presenting better semen. Positive correlations were observed between minor sperm defects and ventral regions of the testes and tails of the epididymis in Girolando bulls. Nelore bulls were less influenced by climatic variables and presented lower temperature in skin surface areas in the infrared thermography examination compared to Girolando bulls. Nelore bulls presented superior semen quality in both fresh and thawed samples than Girolando bulls.

Sex-related differences in sudomotor function in healthy early twenties focused on activated sweat gland density.

The purpose of this study was to quantitatively assess the difference in sudomotor function between healthy males and females in their early twenties by measuring skin surface area and activated sweat gland density (ASGD). The quantitative sudomotor axon reflex test (QSART), a method for evaluating autonomic nervous system activity, was used for quantification. In QSART, the sweat glands are activated directly or indirectly by the subcutaneous application of neurotransmitters, such as acetylcholine, through iontophoresis. This series of mechanisms is called the sudomotor axon reflex. After recording age, height, weight, and several measurements of the forearm, QSART was performed on 101 healthy controls aged 21-26 years to measure ASGD. The mean temperature and humidity on the measurement days were 11.4°C and 58.1% on May 3, 2018, and 14.7°C and 70.3% on May 10, 2018. The result of independent sample t-test showed higher ASGD in women (P < 0.05). The body surface area and the surface area of the forearms were higher in men (P < 0.001), but the number of activated sweat glands was not significantly different according to sex. The activated sweat gland counts of the body and forearms were analyzed through linear regression by age for males and females. Except for the activated sweat gland count of the male body, the analysis showed a tendency to decrease with increasing age but was not statistically significant in any case (P > 0.05). Showing insufficient coefficient of determination (R2), multiple regression analyses with sex and ages did not correct this insignificance between age and activated sweat gland count.

Development of Cost-effective Mosquito Repellent and Distribution Method by Extracting Patchouli Oil

Purpose : We will develop the distribution method of wide oil extract technology to develop repellent using this technology and study mosquito repellent persistence. Research design, data and methodology : A positive control group containing purified water, ethanol and picaridin was prepared, and the experimental control group was prepared in the same proportion as the positive control group, and 0.6% of broad oil was added. The results were summarized using the calculation method according to the avoidance effect and statistically tested by t-test using the excel statistics program. Results : Experiments on skin surface area and voice control of participants showed that men had 8.9% wider skin surface area than women, and voice control tests showed that women were bitten by mosquitoes five times more than men. Both the positive and the experimental control groups had a valid duration of up to three hours, but from the time of five hours, the positive control group had 77% and the experimental control had 90%, indicating a difference of 14.4% over the positive control group. Conclusions : The mosquito repellents developed in this study on the basis of safety and continuity are cost-effective in terms of mosquito repellent, and in addition fragrance, odor removal, perfume, ink, skin care and massage effect.

SkinDot: A modified full-skin transplantation technique

Existing epidermal transplantation procedures applied in burn surgery or wound treatment, such as mesh grafting or the Meek method, do not lead to a restoration of all the skin layers. Dermal skin layers are indispensable in ensuring the quality and function of the transplanted skin as a frictional surface and a carrier of skin appendages such as hair, sweat glands, and sebaceous glands, as well as nerve receptors for detecting pressure, vibration, and temperature. Because of the restricted skin surface area that can be provided by the donor, full-skin transplants cannot be transplanted over a large area. Cultured skin procedures, based on skin cells cultivated in a laboratory, have not yet reached a stage of development where a complex full epidermal transplantation is possible. In particular, the introduction of skin appendages with a functional cell-to-cell communication has not been observed thus far in cultivated skin. Based on the Reverdin transplantation method, in which concave skin islands with epidermal and dermal parts are transplanted, Davis in 1910 described the transplantation of multiple 2−5 mm sized full-skin islands as a new method for the treatment of skin lesions. Further modifying this 100-year-old procedure, we developed a miniaturization and automation of the Davis transplantation method that started in 2011 and called it “SkinDot”. In the following article we describe the effectiveness of the full-skin island transplant procedure in two patients. The transplantation of single 2−3 mm full-skin islands results in a full-skin equivalent without any limits on donor area and with a reduced donor morbidity.

Enhancement of Dermal Delivery of Finasteride Using Microemulsion Systems.

Purpose: Finasteride is a pharmaceutical agent that treats hair loss and acne with hormonal patterns. Due to its poor water solubility, and the smaller surface area in comparison to total skin surface area, penetration of the drug into hair follicles and skin is low. The aim of this research was to formulate, characterize and evaluate in vitro skin permeability of finasteride microemulsions (MEs). Methods: Finasteride MEs were prepared using a pseudo-ternary phase diagram method with an appropriate ratio of oil mixture, surfactant-co-surfactant mixture and water. MEs containing 1% finasteride were prepared with a suitable amount of oily phase and surfactant and cosurfactant. The physicochemical properties of these MEs and in vitro skin permeability of MEs were evaluated. Results: The results showed that the mean droplet size range of ME samples was 5–17 nm and pH was 5.1–5.7. The viscosity of MEs ranged from 86.4–209.6 cps. The drug release profile showed that 49.510% of the drug was released (ME-F-6) over the 24 hours of the experiment. The kinetics of drug release from all selected MEs were approximately described by Higuchi and first-order modeling. All ME formulations with different compositions and properties significantly increased flux and permeability coefficient from rat skin. The selected MEs exhibit 99.9% finasteride after six months of storage. Conclusion: This study showed that any change in the content and composition of MEs could change the physical and chemical properties in addition to ME permeability parameters. The MEs increased permeability of the skin to finasteride.

The Study of Using 3D Scan Technique to Evaluate the Expanding Method of Ear Reconstruction Before Operation

PurposeThree-dimensional scanning technology was used to measure the expansion of the area and size of auricular skin to meet the normal standard of the external ear before ear reconstruction among microtia patients.Materials and MethodsThe skin surface area of microtia patients was measured by three-dimensional scanner: the surface area (S), vertical length (A), vertical curve length (B), transverse length (C), transverse curve length data (D), and then taking the average. Corresponding measurements in healthy adults were also obtained: surface area (S0), the vertical curve length (B0), and transverse curve length (D0) of the normal external ear were obtained by scanning normal adult male ears with reference to the range of the vertical length and the transverse straight length. Mean surface area (S and S0), vertical curve length (B and B0), and transverse curve length (D and D0) were compared between microtia patients and healthy adults.ResultsThe surface area, vertical curve length, and transverse curve length were statistically significantly higher among healthy adults.ConclusionsWith the amount of expanded water injection of 120–130 ml, the expanded skin still does not reach the standard of the normal external ear in terms of skin surface area and size.Level of Evidence IVThis 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.

Long-Term Risk of Skin Cancer Among Childhood Cancer Survivors: A DCOG-LATER Cohort Study.

Skin cancer is common after radiotherapy among childhood cancer survivors (CCSs). We studied risks and risk factors for subsequent skin cancers, with emphasis on radiation dose, exposed skin surface area, and chemotherapeutic agents. The DCOG-LATER cohort study includes 5-year Dutch CCSs diagnosed 1963-2001. Subsequent skin cancers were identified from record linkages with the Netherlands Cancer Registry and Dutch Pathology Registry. Incidence rates were compared with general population rates. Multivariable Cox regression models were used, applying a novel method of case-control sampling enabling use of tumor location in cohort analyses. All statistical tests were two-sided. Among 5843 CCSs, 259 developed 1061 basal cell carcinomas (BCCs) (standardized incidence ratio [SIR] = 29.8, 95% confidence interval [CI] = 26.3 to 33.6; excess absolute risk per 10 000 person-years (EAR) = 24.6), 20 had melanoma (SIR = 2.3, 95% CI = 1.4 to 3.5; EAR = 1.1), and 10 had squamous cell carcinoma (SIR = 7.5, 95% CI = 3.6 to 13.8; EAR = 0.8). Cumulative incidence of BCC 40 years after childhood cancer was 19.1% (95% CI = 16.6 to 21.8%) after radiotherapy vs 0.6% expected based on general population rates. After a first BCC, 46.7% had more BCCs later. BCC risk was associated with any radiotherapy to the skin compartment of interest (hazard ratio [HR] = 14.32, 95% CI = 10.10 to 20.29) and with estimated percentage in-field skin surface area (26-75%: HR = 1.99, 95% CI = 1.24 to 3.20; 76-100%: HR = 2.16, 95% CI = 1.33 to 3.53, vs 1-25% exposed; Ptrend among exposed = .002), but not with prescribed radiation dose and likelihood of sun-exposed skin-area. Of all chemotherapy groups examined, only vinca alkaloids increased BCC risk (HR = 1.54, 95% CI = 1.04 to 2.27). CCSs have a strongly, 30-fold increased BCC risk. BCC risk appears to increase with increasing skin surface area exposed. This knowledge underscores the need for awareness by survivors and their health care providers.

Burn-induced Oxidative Stress and Serum Glutathione Depletion; a Cross Sectional Study.

Several studies have shown the role of oxidative stress in pathophysiology of burn injuries. This study aimed to evaluate the changes of oxidant-antioxidant levels during the week following burn injuries and its correlation with grade of burn. In this prospective cross-sectional study, changes of total glutathione, reduced glutathione (GSH), oxidized GSH (GSSG), GSH/GSSG ratio, as well as Pro-oxidant-antioxidant balance (PAB) were investigated on the 1st, 2nd and 7th days of admission in patients with > 15 % burns. 40 patients with the mean age of 21.1 ± 14.5 were studied (47.5% male). More than 50% of patients were in the 18 - 55 years age range and over 70% had 20% - 60% grade of burn. Total serum glutathione level and GSH had significant decreasing trends (P < 0.001) and GSSG and GSH/GSSG ratio had increasing trends (p < 0.001). No significant correlation was observed between serum GSH level and the total body surface area (TBSA) of burn injury (r = 0.047; p = 0.779). The evaluation of PAB and its correlation with TBSA showed a significant and direct association between them on the 1st (coefficient = 0.516; p = 0.001), 2nd (coefficient = 0.62; p <0.001), and 3rd (coefficient = 0.471; p = 0.002) day of follow up. According to this study, the redox perturbation occurred in burn injury which was measured and proved by decreased GSH/GSSG ratio as well as the shift of PAB in favour of oxidants. Besides, since PAB positively correlated with the severity of dermal damage, it might suggest the application of antioxidants as a part of therapeutic protocol for which the dosage should be proportionate to the surface area of the damaged skin.

The role of interleukins in vitiligo: a systematic review.

Vitiligo is a disorder of the skin that causes depigmentation and asymptomatic macules whose exact cause is still unclear. Although its aetiology is not fully elucidated, the main theory of its pathomechanism is that it is associated with the autoimmune process. There is few summarized information about the role of inflammatory mediators, as interleukins, in vitiligo, so our aim was to present a systematic review of the role of interleukins in vitiligo, focusing on interleukins. In this review, we included all studies assessing interleukin levels in vitiligo patients conducted up to June 2017. Quality assessment of these studies was performed using the Newcastle-Ottawa Scale (NOS). The interleukins mainly involved were IL-2, IL-4, IL-6, IL-10 and IL-17. The studies highlight the crucial role of IL-17 in the onset and progression of the disease, and its synergistic action with IL-2, IL-6 and IL-33. Dysregulated levels of the interleukins were also correlated with the stage of disease, the affected skin surface area, and indicated as the main factor for lymphocyte infiltration found in depigmented regions. These findings illustrate the growing need for new therapies targeting vitiligo and further research into the role of interleukins as an area of particular interest.

Is Sunlight Exposure Enough to Avoid Wintertime Vitamin D Deficiency in United Kingdom Population Groups?

Solar ultraviolet radiation (UVR) is required for cutaneous vitamin D synthesis, and experimental studies have indicated the levels of sun exposure required to avoid a vitamin D deficient status. Our objectives are to examine the sun exposure behaviours of different United Kingdom sectors and to identify if their exposure is enough to maintain winter circulating 25-hydroxyvitamin D above deficiency (>25 nmol/L). Data are from a series of human studies involving >500 volunteers and performed using the same protocols in Greater Manchester, UK (53.5° N) in healthy white Caucasian adolescents and working-age adults (skin type I–IV), healthy South Asian working-age adults (skin type V), and adults with photodermatoses (skin conditions caused or aggravated by cutaneous sun exposure). Long-term monitoring of the spectral ambient UVR of the Manchester metropolitan area facilitates data interpretation. The healthy white populations are exposed to 3% ambient UVR, contrasting with ~1% in South Asians. South Asians and those with photodermatoses wear clothing exposing smaller skin surface area, and South Asians have the lowest oral vitamin D intake of all groups. Sun exposure levels prevent winter vitamin D deficiency in 95% of healthy white adults and 83% of adolescents, while 32% of the photodermatoses group and >90% of the healthy South Asians were deficient. The latter require increased oral vitamin D, whilst their sun exposure provides a tangible contribution and might convey other health benefits.

DESIGN AND PERFORMANCE VERIFICATION OF NEWLY DEVELOPED DISPOSABLE STATIC DIFFUSION CELL FOR DRUG DIFFUSION/PERMEABILITY STUDIES

Objectives: The present study describes a disposable static diffusion cell for in vitro diffusion studies to achieve better results as compared to well existing Franz diffusion cell (FDC) in terms of the absence of bubbles, variable receptor compartment, ease of handling, and faster results.Materials and Methods: The cell consists of a cup-shaped donor compartment made of semi permeable that could be either cellophane membrane or, animal skin fitted to a rigid frame, which is supported on a plastic plate that contains a hole for the sample withdrawal. The receptor compartment is a separate unit, and it could be any container up to 500ml volume capacity. The most preferred receptor compartment is glass beaker. In the present study, goatskin was used as semi-permeable membrane and verification of its performance was carried out through diffusion studies using gel formulations of one each of the four-selected biopharmaceutical classification system (BCS) class drugs. Metronidazole, diclofenac sodium, fluconazole, and sulfadiazine were used as model drugs for BCS Class I, II, III, and IV, respectively.Results: The newly developed diffusion cell (NDDC) was found to provide faster and more reproducible results as compared to FDC. At the time interval of 24 h, the cell was found to exhibit a higher diffusion of metronidazole, diclofenac sodium, fluconazole, and sulfadiazine by 0.65, 0.65, 0.32, and 0.81 folds, respectively. The faster release obtained with NDDC was attributed to a larger surface area of skin as compared to that in FDC.Conclusion: It was concluded that better reproducibility of results could be achieved with NDDC.

3D scanning applications in medical field: A literature-based review

BackgroundMedical field has extensive use of different scanning like X-rays, CT, MRI and Ultrasound. These techniques are quite useful for providing information on the internal organs. However, there is a gap for obtaining information of the outer body parts which can now be taken care by 3D Scanning technologies. A large number of research papers about different scanning techniques and 3D scanning has been studied to identify the impact of them and the subsequent research. The aim of the workDoctors and technologists use the available scanning technologies in different sub-medical fields. This paper endeavours to find out the best possible usage of 3D scanning technologies in the medical area. This paper will help doctors for best treatment to the patient with high knowledge, minimum risks and maximising benefits. Materials and methodsA large number of the relevant research papers from 2008 to April 2018, identified through Scopus are studied using bibliometric analysis, thus to determine strength & limitation and to undertake application analysis. ResultsYear-wise, Journal-wise and relevant applications wise study undertaken in the medical field through Bibliometric research to identify the strength, limitations, and applications of 3D scanning, and other contemporaries scanning technologies. ConclusionThe bibliometric analysis shows that there is an increasing trend in the research work undertaken in 3D scanning application in the medical field, this also explains future potential and contribution. Corporate are exploring 3D scanning for commercial medical applications along with part analysis & designs before the actual production. It helps to create efficient implants easily and quickly. In this paper, we have summarised 3D scanning applications for medical purposes. This technology helps to produce medical implant according to the required specification. It is used for the accurate measuring of patient's body shape, size and skin surface area or an individual part of the body. In medical, data varies from patient to patient, so for producing a 3D digital image, 3D scanning technologies are used which provide digital 3D models. We can obtain improved patient treatment through the integration of digital models with Virtual Reality and Holographic techniques. The significant limitation of this technology is that it can only scan the outer surface of the body or part/model.

An extended Kalman filter for mouse tracking.

Animal tracking is an important tool for observing behavior, which is useful in various research areas. Animal specimens can be tracked using dynamic models and observation models that require several types of data. Tracking mouse has several barriers due to the physical characteristics of the mouse, their unpredictable movement, and cluttered environments. Therefore, we propose a reliable method that uses a detection stage and a tracking stage to successfully track mouse. The detection stage detects the surface area of the mouse skin, and the tracking stage implements an extended Kalman filter to estimate the state variables of a nonlinear model. The changes in the overall shape of the mouse are tracked using an oval-shaped tracking model to estimate the parameters for the ellipse. An experiment is conducted to demonstrate the performance of the proposed tracking algorithm using six video images showing various types of movement, and the ground truth values for synthetic images are compared to the values generated by the tracking algorithm. A conventional manual tracking method is also applied to compare across eight experimenters. Furthermore, the effectiveness of the proposed tracking method is also demonstrated by applying the tracking algorithm with actual images of mouse. Graphical abstract.

Using Terminal Transferase-mediated dUTP Nick End-labelling (TUNEL) and Caspase 3/7 Assays to Measure Epidermal Cell Death in Frogs with Chytridiomycosis.

Amphibians are experiencing a great loss in biodiversity globally and one of the major causes is the infectious disease chytridiomycosis. This disease is caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd), which infects and disrupts frog epidermis; however, pathological changes have not been explicitly characterized. Apoptosis (programmed cell death) can be used by pathogens to damage host tissue, but can also be a host mechanism of disease resistance for pathogen removal. In this study, we quantify epidermal cell death of infected and uninfected animals using two different assays: terminal transferase-mediated dUTP nick end-labelling (TUNEL), and caspase 3/7. Using ventral, dorsal, and thigh skin tissue in the TUNEL assay, we observe cell death in the epidermal cells in situ of clinically infected animals and compare cell death with uninfected animals using fluorescent microscopy. In order to determine how apoptosis levels in the epidermis change over the course of infection we remove toe-tip samples fortnightly over an 8-week period, and use a caspase 3/7 assay with extracted proteins to quantify activity within the samples. We then correlate caspase 3/7 activity with infection load. The TUNEL assay is useful for localization of cell death in situ, but is expensive and time intensive per sample. The caspase 3/7 assay is efficient for large sample sizes and time course experiments. However, because frog toe tip biopsies are small there is limited extract available for sample standardization via protein quantification methods, such as the Bradford assay. Therefore, we suggest estimating skin surface area through photographic analysis of toe biopsies to avoid consuming extracts during sample standardization.

Erythroderma and extensive poikiloderma \u2013 a rare initial presentation of dermatomyositis: a case report

BackgroundDermatomyositis is a humoral-mediated inflammatory myopathy with symmetrical proximal muscle weakness and dermatological manifestations such as Gottron’s papules, heliotrope rash, periungual abnormalities, and flagellate erythema. Erythroderma is a severe and potentially life-threatening dermatological condition with diffuse erythema and scaling involving more than 90% of the skin surface area. Poikiloderma vasculare atrophicans refers to mottled hyperpigmentation and hypopigmentation of the skin with in-between telangiectases and areas of atrophy and is considered a variant of mycosis fungoides. Poikilodermatomyositis is the term given to the condition with poikiloderma and inflammatory myopathy. Only a few cases are reported on erythroderma in dermatomyositis and poikilodermatomyositis. Erythrodermal pattern of dermatomyositis transforming into poikilodermatomyositis is a recognized rare manifestation of dermatomyositis and we could find only one case report in the literature.Case presentationA 53-year-old Sri Lankan woman presented with intermittent fever of 5 months’ duration with erythroderma. Later she developed progressive, symmetrical proximal muscle weakness. Following a short course of small dose steroids, erythroderma settled but changed to extensive poikiloderma involving more than 90% of her skin with her face being relatively spared. She had an early heliotrope rash, shawl sign, and Gottron papules. Electromyography and muscle biopsy were supportive of inflammatory myositis and skin biopsy showed evidence of dermatomyositis. Inflammatory markers and muscle enzymes were also elevated. Autoimmune antibodies and myositis-specific autoantibodies were negative. She was started on orally administered prednisolone 1 mg/kg per day with methotrexate 10 mg once a week and had a good response to treatment with resolution of the skin condition and improvement of muscle power. Imaging studies, endoscopies, and tumor markers did not reveal any malignancy.ConclusionsThis case illustrates a rare presentation of dermatomyositis initially presenting as fever, erythroderma, and proximal muscle weakness and later developing poikiloderma involving more than 90% of the skin. It is important to be aware of this rare presentation to avoid misdiagnosis. With the currently available literature it is not possible to conclude that erythroderma is a bad prognostic factor in dermatomyositis or a predictive factor for a malignancy. Patients have a good response to steroids with a combination of immunosuppressants.

Differences in susceptibility to the skin fluke Benedenia epinepheli between Epinephelus bruneus, E. septemfasciatus, and a new hybrid grouper Kue-Tama, E. bruneus × E. lanceolatus

Interspecific hybrids of marine fish are becoming popular for aquaculture. The advantageous characteristics of hybrid fish include high growth rates, enhanced feed conversion rates, better appearance, high tolerance to environmental stress, and increased resistance to certain diseases. A new hybrid grouper Kue-Tama, female Epinephelus bruneus × male E. lanceolatus has recently been established. This hybrid showed significantly higher growth than that by E. bruneus and is considered a promising new species for aquaculture, especially in temperate areas where the slow growth of E. bruneus has prevented stable farming practices. The present study compared the resistance of Kue-Tama, its maternal parent E. bruneus and E. septemfasciatus, another popular grouper for farming, against the skin fluke, Benedenia epinepheli, which is a problematic parasite in grouper farming. The result of the cohabitation experimental infection showed that E. septemfasciatus was the most resistant to the skin fluke, followed by Kue-Tama and then E. bruneus. There appeared to be a negative relationship between the amount of mucus produced and the parasite intensity suggesting a resistance to skin flukes though the dry-weight of mucus collected from a given skin surface area did not differ between the three groupers. These results demonstrated that Kue-Tama has another favorable characteristic for aquaculture, in addition to their higher growth rate.