Fatores de virulência e susceptibilidade a antifúngicos de Cryptococcus Spp / Virulence factors and susceptibility to Cryptococcus Spp. antifungals

Criptococose é uma doença grave que afeta tanto imunocomprometidos quanto imunocompetentes, com isso analisar a virulência é fundamental para novas terapêuticas. Objetivo: Analisar a capacidade de virulência e susceptibilidade aos antifúngicos de Cryptococcus spp. isolados de líquor de pacientes de hospital do norte do Paraná. Métodos: A partir de dois isolados clínicos C. neoformans e C. gattii, realizou-se a confirmação da identificação. Para a virulência, avaliou-se o tamanho da cápsula, capacidade de sobrevivência após exposição a neutrófilos, produção de melanina e urease. No antifungigrama por difusão em disco utilizou-se: anfotericina B, cetoconazol, voriconazol, itraconazol e miconazol. Resultados: C. gattii destaca-se por maior desenvolvimento da cápsula além da melhor capacidade de sobreviver a fagocitose em relação ao C. neoformans. No antifungigrama, ambos os isolados se apresentam sensíveis às drogas estudadas. Conclusão: Esses achados contribuem para a compreensão das diferentes patogêneses entre C. gattii e C. neoformans.

Cryptococcosis is a serious disease that can affect both immunocompromised and immunocompetent individuals, thus the virulence analysis is fundamental for the development of new treatments. Objective: To analyze the virulence and susceptibility of Cryptococcus spp. isolated from cerebrospinal fluid of patients from a hospital in the north of Paraná. Methods: From two clinical isolates, C. neoformans and C. gattii were confirmed and identified. For virulence, capsule size, survival capacity after exposure to neutrophils, melanin production and urease were evaluated. In the disc-diffusion method, the following antifungals were used: amphotericin B, ketoconazole, voriconazole, itraconazole and miconazole Results: It was observed that C. gattii presents greater results for development of the capsule beside presenting the best ability to survive phagocytosis in relation to C. neoformans. In the disc-diffusion method, both isolates presented sensitivity to the studied drugs. Conclusion: These findings contribute to the understanding of the different pathogens between C. gattii and C. neoformans.

Papilas Fungiformes Pigmentadas de la Lengua. Características Clínicas, Histológicas y Dermatoscópicas de una Serie de Casos Ecuatorianos / Pigmented Fungiform Papillae of the Tongue. Clinical, Histological and Dermatoscopic Characteristics A Serie of Ecuadorian Cases

RESUMEN: Las papilas fungiformes pigmentadas de la lengua, cuyas siglas son PFPT, del inglés Pigmented fungiform papillae of the tongue, es una condición asintomática, no progresiva que se presenta en personas de piel oscura, en las cuales las papilas fungiformes cambian de su color rosado natural, a una gama de café a negro. El objetivo de nuestro estudio es reportar y describir las caractetísticas clínicas, dermatoscópicas e histológicas de la PFPT por primera vez en una serie de pacientes ecuatorianos. Estudio prospectivo simple en el Centro de Especialidades Dermatológicas Garzón, período de dos años. El criterio de inclusión fue cambio de coloración a nivel lingual, se recolectaron datos demográficos, clínicos; fotografías, dermatoscopía, y biopsia, para tinción con hematoxilina-eosina y Fontana-Mason. Examinamos 8.640 pacientres, 15 (12 mujeres, 3 varones) fueron diagnosticados de PFPT. La edad promedio fue 31 años, todos fueron mestizos, con fototipo de piel predominante III y IV. El tiempo de evolución promedio en años fue 5,8. Ninguno tuvo antescedentes familiares o personales relacionados a la patología. La evaluación clínica demostró que el patrón de distribución de acuerdo a la clasificación de Holzwanger en la gran mayoría fue tipo II (13/15). En todos los casos la dermatoscopía y la histología fueron específicas demostrando hallazgos típicos y comprobatorios de PFPT.

ABSTRACT: The pigmented fungiform papillae of the tongue, whose acronyms are PFPT, of the English Pigmented fungiform papillae of the tongue, is an asymptomatic, nonprogressive condition that occurs in dark-skinned people, in which the fungiform papillae change their color natural pink, to a range of brown to black. The aim of our study is to report and describe the clinical, dermatoscopic and histological characteristics of the PFPT for the first time in a series of Ecuadorian patients. A simple prospective study at the Garzón Dermatological Specialty Center, a two-year period. The inclusion criteria was lingual change of color, demographic, clinical data were collected; photographs, dermatoscopy, and biopsy, for staining with hematoxylin-eosin and FontanaMason. We examined 8,640 patients, 15 (12 women, 3 men) were diagnosed with PFTP. The range of age was 31 years, all were mestizos, with skin phototype predominant III and IV. The range of evolution time in years was 5.8. None had family or personal precedents related to the pathology. The clinical evaluation showed that the pattern of distribution according to the Holzwanger classification in the great majority was type II (13/15). In all cases, the dermatoscopy and histology were specific, demonstrating typical and evidential findings of PFPT.

Physiological and lifestyle factors contributing to risk and severity of peri-orbital dark circles in the Brazilian population

AbstractBACKGROUND:Peri-orbital dark circles are a cosmetic concern worldwide, and have been attributed to hyperpigmentation from allergy or atopic dermatitis, blood stasis, structural shadowing effects, and a thin epidermis/dermis under the eye. It is of interest to better understand lifestyle and demographic risk factors and the relative impact of melanin, blood and epidermal/dermal factors on the severity of Peri-orbital dark circles.OBJECTIVE:To compare by non-invasive imaging the impact of biological factors to a visual grading scale for Peri-orbital dark circles, and test the correlation of various demographic factors with Peri-orbital dark circles.METHODS:Subjects completed a lifestyle and health survey, and Peri-orbital dark circles severity was evaluated using standardized photographs. Hyperspectral image analysis was used to assess the contributions of melanin, blood volume, degree of blood oxygen saturation, and dermal scattering.RESULTS:Family history was the most significant risk factor for Peri-orbital dark circles. The average age of onset was 24 years, and earlier onset correlated with higher severity scores. Asthma was significantly associated with Peri-orbital dark circles scores, but self-reported allergy was not. In this study, sleep was not correlated with Peri-orbital dark circles scores. Hyperspectral imaging indicated that melanin was the dominant correlate for Peri-orbital dark circles severity, while oxygen saturation was secondary. The difference between under-eye and cheek measurements for ΔL*and ΔE* were the most significant instrumental parameters correlated with visual assessment of Peri-orbital dark circles severity.CONCLUSION:Although typically associated with lack of sleep, risk of Peri-orbital dark circles is primarily hereditary. The main factors contributing to the appearance of Peri-orbital dark circles are melanin and (deoxygenated) blood.

Efeitos da luz UV-A e visível em células da pele e no cabelo / Effects UV-A and visible light on skin cells and hair

A luz solar apresenta ondas eletromagnéticas em ampla faixa espectral, incluindo as regiões do ultravioleta (UV-C, UV-B, UV-A), visível e infravermelho. Cada região interage com a pele de forma dependente da fotofísica e da fotoquímica dos seus respectivos compostos absorvedores. A luz UV-A causa a geração de espécies reativas de oxigênio e de nitrogênio (EROs e ERNs) através da fotossensibilização de moléculas endógenas (co-enzimas de flavina, porfirinas, melaninas). Quando fotossensibilizadores produzem quantidades de EROs e ERNs maiores do que a capacidade celular de supressão destas espécies, caracteriza-se um quadro de desbalanço redox, que causa lesão em biomoléculas como os ácidos nucleicos, lipídeos e as proteínas. Essas lesões podem levar à morte celular ou a outras transformações fenotípicas e genotípicas e também estimulam a liberação de citocinas pró-inflamatórias. Com a finalidade de melhor compreender a dinâmica dos mecanismos de resposta celular após exposição ao UV-A e ao visível, nós caracterizamos inicialmente as propriedades fotofísicas da melanina e detectamos a produção de oxigênio singlete (1O2) pela fotossensibilização no visível e a supressão desta espécie excitada pela reação do oxigênio singlete com a dupla ligação reativa dos grupos indóis presentes na melanina. Estes processos também foram observados no cabelo e levaram-nos a propor um modelo que explica o efeito da luz visível na estrutura e cor dos cabelos. Demonstramos também que a feomelanina produz mais (30%) 1O2 do que a eumelanina, que sofre maior modificação na sua estrutura por fotodegradação. O efeito destes processos na pele foi estudado a nível celular. Demonstramos que células epiteliais com maior teor de melanina apresentaram maior geração de 1O2 que causa lesão no DNA e morte necro-apoptótica após irradiação com luz visível. A foto-oxidação da melanina pela luz visível nos motivou a estudar um pigmento que fosse foto-protetor não somente contra luz UV-B mas também contra luz visível. A pigmentação com Acetil-Tirosina se mostrou atóxica e protetora contra luz UV-B e visível ao contrário do pigmento com tirosina, que se mostrou protetor do UV-B mas tóxico no visível. Este efeito foi relacionado com a localização celular do polímero e não com a estrutura do mesmo. A luz UV-A, por sua vez, promove o acúmulo de lipofuscina dentro dos vacúolos autofágicos de queratinócitos da pele e que também ativa a fototoxicidade pela luz visível. A lipofuscina dentro dos vacúolos autofágicos é foto-oxidada pela luz visível, causando lesão no DNA e morte celular programada tipo II. Doses UV-A que desencadeiam a liberação de citocinas também foram caracterizados

Sunlight presents electromagnetic radiation over a wide spectral range, including the regions of ultraviolet (UV-C, UV-B, UV-A), visible and infrared. Each region interacts with skin dependending on the photophysics and photochemistry of the respective absorbing compounds. UV-A light causes the generation of reactive oxygen and nitrogen species (ROS and RNS) by photosensitization of endogenous molecules (flavin coenzymes, porphyrins, melanins). When photosensitizers produce amounts of ROS and RNS larger than the cell capacity to suppress these species, a set of redox imbalance, which damages biomolecules such as nucleic acids, lipids and proteins. This damage cause cell death and to other phenotypic and genotypic changes and also stimulates the release of proinflammatory cytokines. In order to better understand the dynamics of the mechanisms of cellular responses after exposure to UV-A and visible light, we initially characterized the photophysical properties of melanin and detected the production of singlet oxygen (1O2) by photosensitization in the visible, as well as the suppression of these excited species by reaction of singlet oxygen with the double bonds of the reactive groups presented in the melanin indols. These processes were also observed in hair and led us to propose a model that explains the effects of visible light on the structure and color of hair. We also demonstrated that pheomelanin produces more (30%) 1O2 than eumelanin, which undergoes a quick change on its structure by photodegradation. The effect of these processes in the skin was studied at the cellular level. We demonstrated that epithelial cells with larger melanin content have stronger generation of 1O2, which causes DNA damage and necro-apoptotic death after irradiation with visible light. The photo-oxidation of melanin by visible light has motivated us to study a pigment that was not only able to protect against UV-B but also against visible. Pigmentation with Acetyl-Tyrosine proved nontoxic and protective against UV-B and visible light instead of pigmentation with Tyrosine, which shielded against UV-B but showed toxicity in the visible. This effect was associated with the polymer, cell location and not with its structure. UV-A light, in turn, promotes the accumulation of lipofuscin, within autophagic vacuoles of keratinocytes also enabling phototoxicity in the visible light. The lipofuscin within the autophagic vacuoles is fotooxidized by visible light, causing DNA damage and programmed cell death type II. Linear dose of UV-A that trigger the release of cytokines were also characterized

Techniques of digital image analysis for histological quantification of melanin / Técnicas de análise de imagem digital para quantificação histológica da melanina

Morphometric analysis of tissue melanin may quantitatively contribute to research on pigmentation disorders. The authors present three methods for image analysis, which allow for identification of melanin-equivalent pixels in the epidermis using Fontana-Masson stain and, therefore, for the calculation of its percentage in the different epidermal layers. Moreover, they discuss the main elements related to the analysis and the need for rigorous standardization of the process.

A análise morfométrica da melanina tecidual pode subsidiar quantitativamente a pesquisa em discromias. Os autores demonstram três técnicas de análise de imagem digital que permitem a identificação dos pixels equivalentes à melanina na epiderme pela coloração de Fontana-Masson, possibilitando o cálculo da sua porcentagem nas diferentes camadas da epiderme, e discutem os principais elementos relacionados à análise e a necessidade de rigorosa padronização do processo.

Retroperitoneal sclerosing PEComa with melanin pigmentation and granulomatous inflammation-A rare association within an uncommon tumor.

PEComa, defined as a perivascular epithelioid cell tumor, displays a wide clinicopathological spectrum. Lately, a sclerosing PEComa has been identified as its distinct variant, but with limited documentation, in view of its rarity. Herein, we describe an uncommon case of a 53-year-old lady, who was referred to us with pain abdomen. Radiological imaging disclosed a well-defined, hypodense retroperitoneal mass. The excised tumor was a round, encapsulated soft tissue mass measuring 7 cm with a tan-brown cut surface. Microscopy showed uniform, epithelioid cells with clear cytoplasm, focal melanin pigmentation and mild nuclear atypia, arranged in sheets and nests around capillary-sized vessels in a dense sclerotic stroma. Additionally, co-existing epithelioid granulomas were noted. On immunohistochemistry (IHC), tumor cells were diffusely positive for HMB45; focally for desmin and smooth muscle actin (SMA), while negative for EMA, CD10, S100-P, Melan A, CD34, AMACR and CK MNF116. This case reinforces sclerosing PEComa as an uncommon, but a distinct clinicopathological entity and exemplifies diagnostic challenge associated with it; necessitating application of IHC markers for its correct identification. Presence of melanin pigment and granulomatous inflammation in the present tumor constitute as novel histopathological findings in a sclerosing PEComa.

Ultrastructural study of the mycelial phase of clinical isolates of Sporothrix schenckii obtained from feline, canine and human cases of sporotrichosis

Using transmission electron microscopy, we studied the presence of melanin and cell wall thickness of clinical isolates of Sporothrix schenckii obtained from cats, dogs and humans as compared to reference strains. We detected differences regarding presence of the melanin among the clinical isolates of S. schenckii and a correlation between presence of melanin and cell wall thickness.

Cor da pele e pigmentos / Skin color and pigments

As características da cor da pele resultam da interação da luz com a epiderme e com a derme. Existem quatro pigmentos importantes na pele normal que afetam a sua cor: melanina, hemoglobina oxigenada, hemoglobina reduzida e vários carotenos.

Comparison of skin erythema and melanin level in sulfur mustard induced chronic skin lesions and normal skin

Sulfur mustard gas is a chemical agent that has been used in many wars, especially in Iran-Iraq war. This chemical agent affects many organs including lungs, eyes and skin, causing numerous acute and chronic lesions including erythema and hyperpigmentation, respectively. This study was conducted to evaluate erythema and melanin in subjects with a history of exposure to sulfur mustard. This case-control study was done on 309 subjects. They were divided into four groups: sulfur mustard-exposed patients with skin lesions [n=87], sulfur mustard-exposed cases without current skin lesions [n=71], non sulfur mustard-exposed patients with dermatitis [n=78] and normal controls [n=74]. Erythema and melanin were measured in 4 areas [forehead, suprasternal, palm and back of hands] by Mexameter MX18 [Courage-Khazaka, Germany]. Erythema was significantly lower in suprasternal and palmar aspect of hands in sulfur mustard-exposed patients with dermatitis [P<0.05] while there was no significant difference in other areas. In terms of melanin, there was a significant difference in the dorsal aspect of hands in all four groups [P<0.05], where patients with dermatitis [both sulfur mustard exposed and normal population] had higher levels of melanin, probably due to pruritus in such areas. Forehead melanin of the normal population was also significantly lower than other three groups [P<0.05] while there was no significant difference between the melanin level of sulfur mustard exposed subjects [with or without dermatitis] and patients with dermatitis. Sulfur mustard contact can affect erythema and melanin content of the skin

Intra-oral hypermelanosis in an HIV sero-positive patient--a case report.

A 32 year old HIV sero positive male patient reported to the Ragas Dental College Annexure at Voluntary Health Services Hospital, Chennai for evaluation of oral health status. He had no history of any drug or tobacco usage. Intra oral examination revealed pigmentation in the buccal mucosa on both sides, which was increasing over the past six months. Histopathologic evaluation of the hyper pigmented lesion was done, using routine Haematoxylin & Eosin and Masson-Fontana stains. The hyper pigmentation was observed to be as a result of increased melanin production in the epithelium. The pathogenesis of hypermelanosis intra-orally in HIV sero-positive patients is discussed in this article.

Oral melanin pigmentation in smoked and smokeless tobacco users in India. Clinico-pathological study.

Tobacco used as smoked and smokeless form induces oral mucosal changes in which intra-oral mucosal pigmentation is one of the clinical manifestations. The melanocyte activity responsible for pigment changes is not well documented in the literature. The present study is undertaken to observe clinical and histological changes in oral buccal and labial mucosa of 41 tobacco users and compared with 8 controls. 95.24% of smokers showed pigmentation of both labial and buccal mucosa. Labial mucosa showed a high degree fo pigmentation (81%) than the buccal mucosa (33.3%). 93.3% of alcoholics showed a high degree of pigmentation. Hypermelanocytosis and melanosis were observed in smokers. Pigmentation at the site of quid placement was absent in smokeless tobacco users but mild pigmentation was observed away from the site of quid placement with the concurrent increase in the number of melanocytes and melanocytic activity.

Constitutive melanin in the cell wall of the etiologic agent of Lobo's disease

Doenca de Jorge Lobo e uma doenca cronica granulomatosa causada por um fungo obrigatorio, em cuja parede celular demonstra-se a presenca de melanina constitutiva. Ao contrario, no P. brasiliensis, e em descricoes de outros ascomicetos filogeneticamente relacionados da familia Onygenaceae, a melanina nao e detectavel pela tecnica de coloracao histoquimica de Fontana-Masson

Projeçöes diencefálicas para o córtex motor e o núcleo pedunculopontino do tegmento, imunorreativas ao hormônio concentrador de melanina (MCH) e/ou ao neuropeptídeo EI (NEI): um estudo com múltiplos traçadores neuronais / Diencephalic origns of melanin-concentrating hormone-and neuropeptide EI - immunoreactive projections to pedunculopontine nucleous and motor cortex: a study with multiple neuronal tracers

O hormônio concentrador de melanina (MCH) é um heptadecapeptídeo cíclico descrito pela primeira vez em hipófise de salmäo e posteriormente em alguns mamíferos. Os corpos celulares McH-ir encontram-se localizados em regiöes específicas do sistema nervoso central (SNC) do rato, como a Zona Incerta (EI) e a Área Hipotalâmica Lateral (AHL) enquanto que as fibras apresentam-se espalhadas por todo o neuro-eixo. Indica que a inervaçäo cortical é densa e homogênea por todo o neocórtex, assim como, a inervaçäo de várias regiöes da formaçäo reticular, como o núcleo pedunculopontino do tegmento (PPTg). Faz um estudo com múltiplos traçadores neuronais em ratos albinos machos (Wistar) usando 6 procedimentos distintos. Utiliza injeçöes de Diamidino Yellow (DY) no córtex motor (CxM) e de True Blue (TB) no PPTg. Demonstra a existência de neurônios MCH-ir marcados com ambos os traçadores distintamente, tanto na ZI quanto na AHL. Realiza controle com leucoaglutimina do Phaseolus vulgaris (PHA-L) injetado nestes territórios, demonstrando a inervaçäo do CxM e do PPTg originários das regiöes onde as células dupla e triplamente marcada foram encontradas. Conclui que estas células säo também NEI-ir; os resultados envidenciam a presença de colaterais de AHL e ZI para 2 regiöes de SNC envolvidas com o comportamento locomotor. Discute a pesquisa em termos de envolvimento do MCH com o comportamento alimentar, que justamente com o locomotor indica uma atuaçäo na busca do alojamento (a maior quantidade de células marcadas säo originárias de fibras localizadas na regiäo motora das vibrissas), e no sistema reticular ativador ascendente

Ausência de melanócitos na epiderme do gambá (Didelphis azarae) adulto / Absence of melanocytes in the epidermis of the adult opossum (Didelphis azarae)

Estudou-se a presença de melanina na epiderme do gambá (Didelphis azarae). Este pigmento näo foi observado histoquimicamente e o estudo ultra-estrutural confirmou a ausência de melanócitos típicos, com ou sem grânulos de melanina

Melanins and lens pigments: a comparative study

Based on previous findings that lens pigments and melanins share many physicochemical properties, human lens pigments and natural (hair) and synthetic melanins were submitted to oxidation with permanganate under strong acidic conditions. This procedure has been utilized for the characterization of melanins and results in the well defined products, thiazole-4,5-dicarboxylic acid (TDCA) and pyrrole-2,3,5-tricarboxylic acid (PTCA), which can be quantitated by high performance liquid chromatography (HPLC). PTCA is regarded as a marker of black eumelanins and was therefore a main component of synthetic DOPA-eumelanin and dark hair. Its identity was established by synthesis from 5-hydroxyindole-2-carboxylic acid. TDCA derives from pheomelanins and was therefore an important component of red hair and synthetic GSH-pheomelanin. TDCA was identified by its retention time relative to PTCA. The analysis of a series of cataract digests of increasing pigmentation (type I < type IV < type V) and a purified fraction of lens pigments (DE52 pigment) revealed the presence in these preparations of both PTCA and TDCA. The concentration of TDCA significantly increased with the degree of pigmentation of the digests and reached a maximum in the DE52 pigment. The TDCA/PTCA ratio was high in the lens preparations and comparable to that given by hair pheomelanin. These findings support that pheomelanin is an integral part of lens pigments. By comparing the yields of TDCA in GSH-pheomelanin and in the purified lens pigment, a 9 per cent contribution of pheomelanin to the lens pigment was estimated

Veriaciones cuantitativas de la melanina durante la muda de pelaje del conejo de los volcanes (Romerologus diazi) / Quantitative variations of metanin during the moulting of atypical fur in the volcano rabbit (Romerolagus diazi)

Se establecen las diferencias cuantitativas de melanina en los diversos estadios del proceso de la muda atípica de pelaje del conejo de los volcanes. La intensidad del color de la piel de los conejos estudiados tuvo relación directa con su contenido de melanina. Las pieles con lunares temporales tuvieron mayor cantidad de melanoide que las normales, y éstas, a su vez, más que las pieles alopécicas