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Mucormycosis is an extremely aggressive fungal disease with a high mortality rate, especially in people with compromised immune systems. Most cases of mucormycosis are caused by the fungus Rhizopus oryzae. The treatments used are based on high doses of antifungals, associated with surgical resections, when it is possible. However, even with this aggressive treatment, the estimated attributable mortality rate is high. There is therefore a need to develop adjuvant treatments. Photodynamic Inactivation (PDI) may be an auxiliary therapeutic option for mucormycosis. Due to the lack of reports in the literature on the morphology and photodynamic inactivation of R. oryzae, characterization of the fungus using Confocal Microscopy and Transmission Electron Microscopy, and different protocols using Photodithazine® (PDZ), a chlorin e6 compound, as a photosensitizer, were performed. The fungus growth rate under different concentrations and incubation times of the photosensitizer and its association with the surfactant Sodium Dodecyl Sulphate (SDS) was evaluated. For the hyphae, both in the light and dark phases, in the protocols using only PDZ, no effective photodynamic response was observed. Meanwhile with the combination of SDS 0.05% and PDZ, inhibition growth rates of 98% and 72% were achieved for the white and black phase, respectively. In the conidia phase, only a 1.7 log10 reduction of the infective spores was observed. High concentration of melanin and the complex and resistant structures, especially at the black phase, results in a high limitation of the PDI inactivation response. The combined use of the SDS resulted in an improved response, when compared to the one obtained with the amphotericin B treatment.

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Most polymeric materials are synthetic and derived from petroleum, hence they accumulate in landfills or the ocean, and recent studies have focused on alternatives to replace them with biodegradable materials from renewable sources. Biodegradable wastes from food and agroindustry, such as spent coffee grounds (SCGs), are annually discarded on a large scale and are rich in organic compounds, such as polysaccharides, that could be used as precursors to produce films. Around 6.5 million tons of SCGs are discarded every year, generating an environmental problem around the world. Therefore, it was the aim of this work to develop films from the SCGs polysaccharide fraction, which is comprised of cellulose, galactomannans and arabinogalactans. Two types of crosslinking were performed: the first forming coordination bonds of calcium ions with polysaccharides; and the second through covalent bonds with 1,4-phenylenediboronic acid (PDBA). The films with Ca2+ ions exhibited a greater barrier to water vapor with a reduction of 44% of water permeability vapor and 26% greater tensile strength than the control film (without crosslinkers). Films crosslinked with PDBA presented 55-81% higher moisture contents, 85-125% greater permeability to water vapor and 67-150% larger elongations at break than the films with Ca2+ ions. Film biodegradability was demonstrated to be affected by the crosslinking density, with the higher the crosslinking density, the longer the time for the film to fully biodegrade. The results are promising and suggest that future research should focus on enhancing the properties of these films to expand the range of possible applications.

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The objective was to evaluate the effect of dietary 25-hydroxycholecalciferol (25(OH)D3) or vitamin D3 (VitD3) and different total calcium (Ca) levels on the performance, carcass characteristics, blood, enzymatic, and bone biochemistry of broilers reared under heat stress between 1 and 42 days of age. A total of 504 male, Cobb 500, broiler chickens were distributed in a completely randomized design in a 2 × 4 factorial arrangement (VitD3 or 25(OH)D3 × four Ca levels (100, 90, 80 and 70% of the recommendations of Rostagno et al. (2011)), eight treatments, seven replicates and nine broilers per cage. Feed intake and feed conversion ratio did not (p>0.05) vary when levels of Ca were reduced and vitamin D3 sources were supplemented in the diets from 1 to 21 days for broilers chickens. 25 (OH)D3 increased weight gain results (p<0.05). From 1 to 42 d, no differences (p>0.05) were observed on performance, carcass yields and meat quality, bone deposition of Ca and P, and alkaline phosphatase concentration. Higher serum (p<0.05) concentrations of Ca and P were found in broilers fed with 25(OH)D3. The replacement of VitD3 with 25(OH)D3 and the Ca reduction of 30% in diets did not negatively affect performance, carcass characteristics, and Ca and P deposition in the tibia of broilers at 42 days of age, under heat stress.(AU)

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Characterization of phases of soft matter systems is a challenge faced in many physical chemical problems. For polymorphic fluids it is an even greater challenge. Specifically, glass forming fluids, as water, can have, besides solid polymorphism, more than one liquid and glassy phases, and even a liquid-liquid critical point. In this sense, we apply a neural network algorithm to analyze the phase behavior of a mixture of core-softened fluids that interact through the continuous-shouldered well (CSW) potential, which have liquid polymorphism and liquid-liquid critical points, similar to water. We also apply the neural network to mixtures of CSW fluids and core-softened alcohols models. We combine and expand methods based on bond-orientational order parameters to study mixtures, applied to mixtures of hardcore fluids and to supercooled water, to include longer range coordination shells. With this, the trained neural network was able to properly predict the crystalline solid phases, the fluid phases and the amorphous phase for the pure CSW and CSW-alcohols mixtures with high efficiency. More than this, information about the phase populations, obtained from the network approach, can help verify if the phase transition is continuous or discontinuous, and also to interpret how the metastable amorphous region spreads along the stable high density fluid phase. These findings help to understand the behavior of supercooled polymorphic fluids and extend the comprehension of how amphiphilic solutes affect the phases behavior.

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Docosahexaenoic acid (DHA, 22:6n-3) is an essential long chain polyunsaturated fatty acid associated with the development of the nervous system that has to be consumed by infants through breast milk or complementary food sources and which consumption is also usually inadequate in preschoolers. In this work, the in vitro bioaccessibility of DHA from two commercial infant formulas (8.9 and 9.1%) and two preschool children milks (6.9 and 7.2%), with similar DHA contents but formulated with different ingredients, was not improved by the presence of egg phospholipids in the product formulation. In addition, the importance of the choice of an age-appropriate in vitro digestion method was demonstrated by comparing the DHA bioaccessibility from the infant formulas by the Infogest 2.0 standardized method and a simulated digestion method specific for infants.

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Ocular toxoplasmosis is the major cause of infectious posterior uveitis worldwide, inducing visual field defect and/or blindness. Despite the severity of this disease, an effective treatment is still lacking. In this study, spiramycin-loaded PLGA implants were developed aiming at the treatment of ocular toxoplasmosis. Implants were manufactured by a hot-molding technique, characterized by Fourier Transform Infrared Spectroscopy, X-Ray Diffraction, Differential Scanning Calorimetry, Scanning Electron Microscopy; evaluated in terms of ocular biocompatibility by immunofluorescence, flow cytometry, cell migration, Hen's egg test-chorioallantoic membrane (HET-CAM) irritation test; and investigated in terms of in vitro efficacy against Toxoplasma gondii . Characterization techniques indicated that spiramycin was dispersed into the polymeric chains and both substances preserved their physical structures in implants. The HET-CAM test indicated that implants did not induce hemorrhage or coagulation, being non-irritant to the CAM. ARPE-19 cells showed viability by MTT assay, and normality in cell cycle kinetics and morphology, without stimulating cell death by apoptosis. Finally, they were highly effective against intracellular parasites without inducing human retinal pigment epithelial cell death. In conclusion, spiramycin-loaded PLGA implants represent a promising therapeutic alternative for the local treatment of ocular toxoplasmosis.

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Resumo Enquadramento: As desigualdades sociais manifestam-se como um desafio contemporâneo e os profissionais de enfermagem possuem um papel importante no seu combate, na defesa da justiça social e na promoção de relações de poder mais democráticas. Objetivo: Analisar as dimensões das desigualdades sociais nos discursos de entidades representativas da enfermagem brasileira. Metodologia: Estudo qualitativo, do tipo documental, que utilizou 10 documentos de posicionamento público de 3 sociedades da enfermagem brasileira. Resultados: O discurso sobre desigualdades sociais é representado pelas dimensões Género, Raça/Etnia, Fator económico e Outras vulnerabilidades, evidenciando a naturalização do racismo e da discriminação como condição que afeta a sociedade e a enfermagem. Os interesses específicos da enfermagem são apresentados como interesses gerais da sociedade, sobretudo quanto às dimensões de desigualdades de género e raça/etnia. Denúncia, defesa de públicos vulnerabilizados e persuasão dos atores sociais são os principais modos de organização discursiva presentes no posicionamento das entidades. Conclusão: Há um alto comprometimento discursivo das entidades com as desigualdades de grupos sociais, nomeadamente no que concerne ao género, raça/etnia, fator económico e outras vulnerabilidades, num posicionamento político de justiça social.

Abstract Background: Social inequalities are a contemporary challenge. Nurses play an important role in fighting against them, defending social justice and promoting more democratic power relations. Objective: To analyze the dimensions of social inequalities in the discourses of representing entities in Brazilian nursing. Methodology: Qualitative document study in a sample of ten public texts illustrating the positioning of 3 Brazilian nursing entities. Results: The discourse on social inequalities is represented by the dimensions of Gender, Race/Ethnicity, Economic status, and Other vulnerabilities, highlighting the normalization of racism and discrimination as factors that affect society and nursing. Nursing's specific interests are presented as society's general interests, particularly regarding the dimensions of gender-based and racial inequalities. The identification and defense of vulnerable groups and persuasion of social actors are the main forms of discourse organization in the entities' position. Conclusion: Representing entities are highly committed to a discourse on social inequalities, namely gender-based, racial, related to economic status and other vulnerabilities.

Resumen Marco contextual: Las desigualdades sociales son un desafío contemporáneo y los profesionales de la enfermería tienen un papel importante que desempeñar para combatirlas, defender la justicia social y promover relaciones de poder más democráticas. Objetivo: Analizar las dimensiones de las desigualdades sociales en los discursos de las entidades representativas de la enfermería brasileña. Metodología: Estudio cualitativo de tipo documental, que utilizó 10 documentos de posicionamiento público de 3 sociedades de enfermería brasileñas. Resultados: El discurso sobre las desigualdades sociales está representado por las dimensiones de Género, Raza/Etnia, Factor económico y Otras vulnerabilidades, lo que destaca la naturalización del racismo y de la discriminación como condición que afecta a la sociedad y la enfermería. Los intereses específicos de la enfermería se presentan como intereses generales de la sociedad, especialmente en las dimensiones de desigualdades de género y raza/etnia. La denuncia, la defensa de los grupos vulnerabilizados y la persuasión de los actores sociales son los principales modos de organización discursiva presentes en el posicionamiento de las entidades. Conclusión: Existe un alto compromiso discursivo de las entidades con las desigualdades de los grupos sociales, en particular, en relación con el género, la raza/etnia, el factor económico y otras vulnerabilidades, en un posicionamiento político de justicia social.

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Lidocaine is a versatile drug that not only provides local anesthesia, but also reduces anesthetic requirements of other agents and has antiarrhythmic, pro-kinetic, anti-inflammatory, antiendotoxemic and antioxidant effects. As it is a drug commonly used in critically ill patients, its safety from the cardiovascular system should be ensured. The aim of this study was to determine the effects of a continuous rate infusion (CRI) of lidocaine on left ventricular systolic and diastolic function of healthy rabbits sedated with midazolam by use of transthoracic echocardiography. Ten New Zealand healthy rabbits were sedated with intramuscular midazolam (1 mg/kg) and enrolled in two experimental treatments (control or lidocaine). The control treatment (CT) comprised an intravenous bolus of 0.9% sodium chloride (0.05 mL/kg) followed by CRI at 5 mL/h, whereas the lidocaine treatment (LT) comprised a bolus of 2% lidocaine without epinephrine at 1 mg/kg followed by CRI at 50 µg/kg/minute. Echocardiographic and hemodynamic variables were studied. Variables were recorded at baseline (TB) and 20, 40 and 60 minutes following start of CRI (T20, T40 and T60, respectively). No differences were found between treatments. The results of this study demonstrate that a continuous rate infusion of lidocaine at 50 µg/kg/minute does not impair echocardiographic indices of left ventricular systolic and diastolic function of healthy rabbits sedated with midazolam.

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The fractional derivative concept to treat non-isothermal solid state thermal decomposition was employed in this work. Simulated data were compared with the exact solutions for the method validation. Calculated fractional kinetics data for four heating rates were initially considered and the Kissinger-Akahira-Sunose (KAS) method demonstrate that, although the activation energy is not retrieved, it can be useful to determine a single or multistep process. Experimental thermal decomposition data of lumefantrine heated at 5, 10 ,15, and 20 oC min- 1 were fitted for a single-step process. The kinetic parameters were retrieved for integer and fractional kinetics, considering some ideal and general models. Application of the KAS method to these data demonstrated an activation energy dependent on the conversion rate, indicating a multistep process. Five data subintervals were fitted separately using the general model with variable derivative order. It was found a process that occours with integer order derivative until α = 0.3 and fractional order for α > 0.3 with combination of simultaneous reactions, since the parameters do not correspond to any ideal model. The determined activation energies showed the same increasing behavior observed in the KAS approach. The results for multistep process presented an error 102 times smaller if compared with the best result, considering a single-step process. Therefore, the fractional kinetic model presents a powerful extension to the usual thermal data analysis.

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Human spermatogonial stem cells (SSCs) are an essential source to maintain spermatogenesis as an efficient process for daily sperm production with high self-renewal capacity along adulthood. However, the phenotype and the subpopulation that represent the real reserve SSC for the human testis remain unknown. Moreover, although SSC markers have been described for undifferentiated spermatogonia (Adark and Apale), the existence of a specific subtype that could be identified as the actual/true SSC has not yet been fully determined. Herein we evaluated spermatogonial morphology, kinetics, positioning regarding blood vasculature in relation to protein expression (UTF1, GFRA1, and KIT) as well as proliferative activity (MCM7) and identified a small subpopulation of Adark with nuclear rarefaction zone (AdVac) that behaves as the human reserve SSC. We show that AdVac is the smallest human spermatogonial population (10%), staying quiescent (89%) and positioned close to blood vessels throughout most of the stages of the seminiferous epithelium cycle (SEC) and divides only at stages I and II. Within this AdVac population, we found a smaller pool (2% of A undifferentiated spermatogonia) of entirely quiescent cells exhibiting a high expression of UTF1 and lacking GFRA1. This finding suggests them as the real human reserve SSC (AdVac UTF1+/GFRA1-/MCM7-). Additionally, Adark without nuclear vacuole (AdNoVac) and Apale have similar kinetic and high proliferative capacity throughout the SEC (47%), indicating that they are actively dividing undifferentiated spermatogonia. Identification of human stem cells with evident reserve SSC functionality may help further studies intending to sort SSCs to treat male diseases and infertility.

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Com o presente estudo, objetivou-se avaliar o tempo de fornecimento de náuplios de artêmia e o período de transição alimentar para pós-larvas de acará-severo (Heros severus). Foram utilizadas 450 pós-larvas de acará-severo, distribuídas em 45 aquários, em delineamento inteiramente ao acaso, em esquema fatorial 3x5, com três repetições. Foram avaliados três períodos de fornecimento de náuplios de artêmia: zero, cinco e 10 dias. Além disso, foram testados cinco períodos de transição alimentar: um, dois, três, quatro e cinco dias, quando os animais receberam alimentação conjunta de náuplios de artêmia e dieta farelada. Ao final do experimento, os peixes foram contados, pesados e fotografados, para avaliação do desempenho produtivo e das variáveis morfométricas. Para todos os parâmetros avaliados, não foi constatada interação entre o tempo de fornecimento de artêmia e o período de transição alimentar. Os animais alimentados por 10 dias com náuplios de artêmia apresentaram os melhores resultados de desempenho e as maiores variáveis morfométricas. Os peixes que receberam alimentação conjunta por três dias apresentaram desenvolvimento satisfatório. Dessa forma, recomendam-se 10 dias de fornecimento de náuplios de artêmia e três dias de transição alimentar, antes de se iniciar o fornecimento exclusivo de dieta inerte para pós-larvas de acará-severo.(AU)

The aim of this study was to evaluate the supply time of artemia and the period of food transition to post-larvae of severum (Heros severus). A total of 450 post-larvae of severum were distributed in 45 aquaria, in a completely randomized design, in a 3x5 factorial scheme, with three replicates. Three feeding periods with artemia nauplii were evaluated: zero, five, and ten days. In addition, five feeding transitions were tested: one, two, three, four and five days, when the animals received joint feeding of artemia nauplii and dry diet. At the end of the experiment the fish were counted, weighed and photographed, to evaluate the productive performance and the morphometric variables. For all parameters evaluated, no interaction between the time of artemia supply and the feeding transition period was observed. Animals fed for 10 days with artemia nauplii showed the best performance results and the highest morphometric variables. The fish that received joint feeding for three days presented satisfactory development. Thus, 10 days of artemia nauplii supply and three days of feeding transition is recommended before starting the exclusive supply of diet inert to post-larvae of severum.(AU)

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Com o presente estudo, objetivou-se avaliar o tempo de fornecimento de náuplios de artêmia e o período de transição alimentar para pós-larvas de acará-severo (Heros severus). Foram utilizadas 450 pós-larvas de acará-severo, distribuídas em 45 aquários, em delineamento inteiramente ao acaso, em esquema fatorial 3x5, com três repetições. Foram avaliados três períodos de fornecimento de náuplios de artêmia: zero, cinco e 10 dias. Além disso, foram testados cinco períodos de transição alimentar: um, dois, três, quatro e cinco dias, quando os animais receberam alimentação conjunta de náuplios de artêmia e dieta farelada. Ao final do experimento, os peixes foram contados, pesados e fotografados, para avaliação do desempenho produtivo e das variáveis morfométricas. Para todos os parâmetros avaliados, não foi constatada interação entre o tempo de fornecimento de artêmia e o período de transição alimentar. Os animais alimentados por 10 dias com náuplios de artêmia apresentaram os melhores resultados de desempenho e as maiores variáveis morfométricas. Os peixes que receberam alimentação conjunta por três dias apresentaram desenvolvimento satisfatório. Dessa forma, recomendam-se 10 dias de fornecimento de náuplios de artêmia e três dias de transição alimentar, antes de se iniciar o fornecimento exclusivo de dieta inerte para pós-larvas de acará-severo.(AU)

The aim of this study was to evaluate the supply time of artemia and the period of food transition to post-larvae of severum (Heros severus). A total of 450 post-larvae of severum were distributed in 45 aquaria, in a completely randomized design, in a 3x5 factorial scheme, with three replicates. Three feeding periods with artemia nauplii were evaluated: zero, five, and ten days. In addition, five feeding transitions were tested: one, two, three, four and five days, when the animals received joint feeding of artemia nauplii and dry diet. At the end of the experiment the fish were counted, weighed and photographed, to evaluate the productive performance and the morphometric variables. For all parameters evaluated, no interaction between the time of artemia supply and the feeding transition period was observed. Animals fed for 10 days with artemia nauplii showed the best performance results and the highest morphometric variables. The fish that received joint feeding for three days presented satisfactory development. Thus, 10 days of artemia nauplii supply and three days of feeding transition is recommended before starting the exclusive supply of diet inert to post-larvae of severum.(AU)

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BACKGROUND: Bioengineering aims to develop innovative scaffolds to improve cellular activities for tissue regeneration. OBJECTIVES: To evaluate the biological behavior of human dental pulp stem cells (hDPSCs) seeded onto an experimental polymeric-based scaffold comprising poly-ε-caprolactone/poly (rotaxane). MATERIAL AND METHODS: Adhesion, viability, and proliferation as well as alkaline phosphatase (ALP) activity, mineralized nodule formation (alizarin red assay), and expression of genes related to osteogenic differentiation, including ALP, type 1 collagen alpha 1 (COL1A1), Runt-related transcription factor (Runx-2), and osteocalcin (BGLAP/OCN), were evaluated in hDPSCs seeded onto polymeric scaffolds. RESULTS: hDPSCs expressed typical levels of mesenchymal stem cell surface markers. Cell growth increased upon cultivation on polymeric blend scaffold and the cells gained osteoblast-like appearance. Fourteen days after seeding hDPSCs on the scaffolds, irrespective to the culture medium used (clonogenic or mineralization medium), the cells presented ALP activity higher than that of control cells grown in clonogenic medium. The cells cultivated in mineralization medium on the scaffold showed significantly higher expression of all genes than the control cells, except for BGLAP gene expression. At 21 days, the group cultivated on the scaffold and mineralization medium showed maximum level of mineralization. SIGNIFICANCE: Poly-ε-caprolactone/poly (rotaxane) blend is noncytotoxic to hDPSCs and improved genomic and functional osteogenic differentiation. Thus, poly-ε-caprolactone/poly (rotaxane) blend may serve as a promising bioactive biomaterial for bone tissue bioengineering.

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This study analyzes the effects of the Rapha® system, which employs red light therapy (RLT) and a natural latex biomembrane in the healing of neuropathic ulcers associated with the diabetic foot. We conducted a randomized, controlled, blinded clinical trial with 15 participants that were divided into three groups (GI, GII and GIII): (i) Rapha® system application by the participant and a health professional at home, with clinical status evaluation every 2 weeks at the hospital (GI); (ii) standard protocol used in Brazil, performed by a health professional at the hospital (GII; control); and (iii) the Rapha® system applied by the participant at home and clinical status evaluation every 2 weeks at the hospital (GIII). We used image processing techniques on photographic recordings of the lesions, and several statistical tests were used to analyze the data, allowing for the comparison of the average results for all groups. The average healing rates of GI, GII, and GIII were 77.0, 51.4, and 80%, respectively. The granulation tissue evaluation indicated a higher efficacy in the tissue repair of lesions treated with the Rapha® system. In conclusion, the Rapha® system proved to be an effective healing system, even when self-applied at the patient's home.

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OBJECTIVE: The aim of this systematic review was to evaluate the most appropriate hydrogel scaffold type (natural, synthetic or hybrid) to be applied with stem cells for dental pulp regeneration. The findings should help clinicians make an informed choice about the appropriate scaffold to be applied for this approach. DESIGN: Three electronic databases were searched (Medline, Web of Science and Scopus). The review was conducted based on the Preferred Reporting Items for Systematic Reviews and Meta-analysis (PRISMA). RESULTS: From 4990 potentially relevant studies initially identified, 18 papers fulfilled the eligibility criteria and were considered for this review. Natural scaffolds were applied in most studies. Collagen was the most studied scaffold. In 5 of 10 studies, only growth factors were added to the constructs. Even without growth factors, these scaffolds containing stem cells were able to support the formation of dentin. The synthetic scaffolds were the least studied. Only 4 studies were selected, and in 3 of them, the same scaffold (Puramatrix) was evaluated. Puramatrix by itself was unable to form dental pulp when dental pulp stem cells were not present. Synthetic and hybrid hydrogels were unable to attract stem cells from the host. The presence of growth factors in these constructs seems to be of relevance since dental pulp tissue formation was achieved only when the hybrid scaffold was applied with growth factors. CONCLUSION: All types of hydrogel-based scaffolds, when containing mesenchymal stem cells, are able to form connective tissue with different degrees of similarity to dental pulp. However, current data is too heterogeneous to compare and identify the advantages of any specific scaffold.

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In light of the scarcity of information about the occurrence and epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) and coagulase-negative staphylococci (MRCNS) in small ruminants in general, and particularly dairy goats, we launched this limited-scope study. The findings reported here show the detection of MRSA and MRCNS in goat milk and teat skin samples from dairy goat herds in the state of Ohio. A total of 120 milk samples and 120 teat-swab samples were collected from 5 farms. After conventional isolation and phenotypic characterization of the staphylococci colonies, bacterial isolates were tested by PCR assay targeting the genes nuc to identify Staphylococcus aureus and mecA to detect MRSA and MRCNS. The clonal complexes of MRSA isolates was also determined by multiloccus sequence typing. Fifteen (6.2%) positive S. aureus samples were found in this study: 9 from milk and 6 from teat skin samples. Four (2%) MRSA isolates were detected and, using multiloccus sequence typing genotyping, these were designated to clonal complexes CC133 (n = 2; milk samples) and CC5 (n = 2; teat skin). Three (1.25%) coagulase-negative staphylococci isolates from the teat skin also harbored the mecA gene. Although, the MRSA isolated from milk samples is not a typical human-associated lineage, the CC5 clone isolated from teat skin is a common and widespread clonal complex associated with humans, suggesting that this extramammary niche could be a relevant reservoir of methicillin-resistant staphylococci. Furthermore, the fact that 75% of MRSA were recovered from 1 farm showing poor hygiene practices strengthens the hypothesis that good hygiene practices could be useful to prevent persistence and spread of MRSA at a farm level.

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Amazon mosses, such as Holomitriopsis laevifolia and Leucobryum sp. are naturally exposed to high levels of solar ultraviolet (UV) radiation. Theoretically, under environmental stress conditions these mosses have developed protective chemical and metabolic strategies against UV damage, by way of biosynthesis of secondary metabolites, such as flavonoids. The present paper aimed to evaluate the free-radical scavenging activity, and the photoprotective, mutagenic and photomutagenic potencies of the methanolic (ME), aqueous (AE), hydroalcoholic (HE), ethanolic (EE) extracts of H. laevifolia and Leucobryum sp. The phenolic contents were evaluated by spectrophotometry and by High-Performance Liquid Chromatography (HPLC). The present findings showed that the AE and HE of H. laevifolia and the AE of Leucobryum sp. presented the highest phenolic contents. The HPLC analysis indicated the presence mainly of phenolic and cinnamic acids, flavonols, flavones and flavanones. The AE and EE of H. laevifolia and the AE and HE of Leucobryum sp. efficiently scavenged the 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical. All extracts showed significant values of in vitro Sun Protection Factor alone, and HE of Leucobryum sp. showed a synergistic effect in association with benzophenone-3. None of the extracts induced mutagenicity in the auxotrophic strains for histidine of Salmonella typhimurium, and photomutagenicity of the TA102 and TA104 strains was not detected after exposure to UV-A radiation. Besides, all extracts showed photoprotective activity against UV-A radiation for the TA104 strain, including synergistic protection in association with BP-3. Thus, the constituents in H. Laevifolia and Leucobryum sp. could be good candidates for cosmetic and dermatological applications, particularly in association with synthetic UV filters, since the concentration of the filters in the final product could be reduced.

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BACKGROUND: The physiopathology of epidermal hypermelanization in melasma is not completely understood. Several cytokines and growth factors are increased in skin with melasma, nevertheless, nor the pathways involved in the increased αMSH expression have been adequately evaluated, nor a model for sustained focal melanogenesis is available. OBJECTIVE: To explore stimulatory pathways for epidermal pigmentation in facial melasma related to αMSH: those linked to ultraviolet radiation, oxidative stress, inflammation, neural crest pigmentation cell differentiation and antagonism of αMSH. METHODS: Paired skin biopsies (3 mm) from 26 women with facial melasma and from normal adjacent skin (<2 cm far) were processed for immunofluorescence with markers for p53, p38, αMSH, MC1R, Melan-A, IL-1α, COX2, Wnt1, WIF-1 and ASIP. RESULTS: The fluorescence intensity in the skin from melasma was higher for MC1R, αMSH at epidermis as at melanocytes (P < 0.05). There were no differences between the sites in epidermal protein expression of COX2, IL-1α, p53, WIF-1 and ASIP (P > 0.1). P53 was expressed only in epidermis, without difference between sites (P = 0.92). WNT1 was remarkable in the epidermis of melasma (P < 0.01), but not in dermis. Positive p38 cells were prominent in the upper dermis of melasma (P < 0.01), despite no marking in epidermis. CONCLUSION: Melanogenesis in melasma involves epithelial secretion of αMSH and activation of the Wnt pathway; nevertheless, it seems to be independent of the stimulation by ultraviolet radiation/p53, IL-1α, COX2/PgE2 , WIF-1 and ASIP. Damaged cells at upper dermis suggest the role of senescence/autophagy in sustained pigmentation in melasma.

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