Navegando por Assunto "Cromoblastomicose"

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Análise da interação in vitro entre Fonsecaea pedrosoi e macrofagos peritoneais de camundongos C57/BL6 e BALB/c
(Universidade Federal do Pará, 2008-04-02) YAMANO, Suellen Sirleide Pereira; SALGADO, Claudio Guedes; http://lattes.cnpq.br/2310734509396125
Chromoblastomycosis (CBM) is a chronic, subcutaneous, granulomatous infection caused by traumatic implantation in the skin of several dematiaceous fungi, usually Fonsecaea pedrosoi. Brazil has second highest disease prevalence in the world and Para State is the most endemic area. Histologically, CBM is characterized by the presence of multinucleated giant cells and sclerotic cells can be found engulfed by macrophages. The objective of this study was to analyze the different aspects of interaction between peritoneal macrophages from BALB/c or C57/BL6 mice with F. pedrosoi conidia or sclerotic cells, calculating infection, phagocytosis and cellular fusion rates. The results showed phagocytosis and infection rates with conidia higher than sclerotic cells to BALB/c (p <0.05), while the rate of cellular fusion was higher for sclerotic cells interaction, with Langhans giant cells formation, in comparison to foreign-body giant cells after interaction with conidia. Macrophages from BALB/c co-cultured with conidia produced more TNF-α than control group after 3 to 72 hours, and more IL-10 after 3h. Macrophages interacting with sclerotic cells produced more TNF-α than control group after 1h and 3h, and the amount of IL-10 was higher after 72h of interaction. In the co-culture of C57/BL6 macrophages with conidia, the presence of large vacuoles after 24h was observed, while in the coculture with sclerotic cells, macrophages were detached from coverslip glasses after 24 h. Our results indicate higher levels of TNF-α after interaction of conidia compared to controls at 1 and 72 h and increase of IL-10 after 48h. However, after interaction with sclerotic cells, only IL-10 differed from control, being higher after 1 to 48 hours. All of these data suggest that macrophage response to fungus is different between BALB/c and C57/BL6 mice, differing also on the response of the same type macrophage for each fungal form, sclerotic cells apparently being more immunogenic than conidia.
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Análise morfológica in vitro da ação de antifúngicos em cepas de Fonsecaea pedrosoi
(Universidade Federal do Pará, 2014-02-21) MASSOUD JUNIOR, Heleno Ramos; SALGADO, Claudio Guedes; http://lattes.cnpq.br/2310734509396125
Choromoblastomycosis (CBM) is a disease caused by traumatic implantation of many species of melanized fungi. The State of Pará is the major endemic area in Brazil and Fonsecaea pedrosoi is the major etiological agent. The treatment is not standardized and many forms of interventions are related in the literature. In the other hand, the in vitro susceptibility test to antifungal drugs may help in the therapeutic choice and in the identification of resistant strains. The objective of this work is to evaluate the in vitro susceptibility of 20 F. pedrosoi clinical isolates to itraconazole (ITZ), ketoconazole (KCZ), fluconazole (FCZ) and terbinafine (TBF) as well as the possible morphological alterations induced by ITZ or TBF in the Minimal Inhibitory Concentration (MIC) and high concentrations. The tests were performed according to the Clinical and Laboratory Standards Institute (CLSI, M38-A2 document) recommendations. The final concentrations of ITZ, TBF and KCZ in each test were to 16 to 0.03 μg/mL. To FCZ the final concentrations were to 64 to 0.125 μg/mL. The MIC was defined as the lowest drug concentration that inhibit 100% the visual growth when compared to the non-treated group after five days of incubation at 30°C. ITZ proved to be the most effective drug in vitro against F. pedrosoi (CIM 90= 1μg/mL). TBF showed a low drug activity with 70% of the isolates with MIC ≥ 0.5 μg/mL. The conidia morphological analysis revealed an increasing in the diameter, an interruption of the cellular division and the formation of little chains after the treatment with ITZ in the MIC. At the high concentration used in the susceptibility test we noticed an irregular shape, a detachment of pigmented material from the cell wall and a vacuolization. Rupture in cell wall and amorphous conidia were observed at 32 μg/mL and 64 μg/mL. Significant alterations were not observed after treatment with TBF at the same concentrations. Moreover, the 5-fluorocytocise (5-FC) and FCZ do not stop the conidia growth at high concentrations. However, ultrastructure alterations were noticed after treatment with 5-FC 64 μg/mL. Thus, it is suggested a different morphological pattern after ITZ or TBF treatment during the in vitro susceptibility test. In synthesis, ITZ shown better in vitro antifungal activity while 5-FC only provoked structures alterations in the highest concentration tested.
Acesso aberto (Open Access)
Avaliação da suscetibilidade in vitro de Fonsecaea spp a diferentes antifúngicos e análise das concentrações plasmáticas de itraconazol e hidroxiitraconazol em pacientes com cromoblastomicose na região amazônica
(Universidade Federal do Pará, 2015-06-18) GRISÓLIA, Daniella Paternostro de Araújo; SALGADO, Claudio Guedes; http://lattes.cnpq.br/2310734509396125
Chromoblastomycosis (CBM) is a fungal infection by implantation in the skin, with a chronic evolution, caused by traumatic inoculation of dematiaceous fungi. Pará state is one of the most important endemic areas in Brazil, and F. pedrosoi is the most prevalent agent. The low cure rate is consequence of the lack of a standardized treatment regimen and there are a few works related to susceptibility of black fungi to the drugs available for treatment. The main objective of this study was to evaluate in vitro susceptibility of clinical isolates of Fonsecaea spp. against nine antifungals, correlating itraconazole (ITZ) minimal inhibitory concentrations (MIC) and minimal fungicidal concentrations (MFC) with clinical outcome and the types of lesions. Furthermore, plasma levels of ITZ and ITZOH were measured in patients treated in Dr Marcello Candia Reference Unit in Sanitary Dermatology (UREMC, Pará State). Thirty-eight clinical isolates of Fonsecaea spp. were evaluated against ITZ, ketoconazole (CTZ), posaconazole (PCZ), voriconazole (VCZ), fluconazole (FCZ), amphotericin B (ANF B), caspofungin (CAS), terbinafine (TBF) and 5-fluorocytosine (5-FLU), according to the protocol of the Clinical and Laboratory Standards Institute (CLSI, M38-A2 document). After 5 days of incubation at 30°C, MICs were determined visually evaluating fungal growth in different drug concentrations, in comparison to the drug-free control wells. PCZ was the most effective (MIC 0.28 μg/ml, MFC 4.35 μg/ml) drug. The correlation between the MIC to ITZ and clinical outcome results demonstrate that patients with worsening had a media value ± SEM (0.90 ± 0.10 μg/ml) superior to no improvement (0.45 ± 0.05 μg/ml) or improvement (0.59 ± 0.05 μg/ml) and cure (0.45 ± 0.05 μg/ml), with a significant difference (p < 0.05) among the groups. There was no correlation between ITZ CIM and types of lesions. We observed a large inter-individual variability of ITZ and ITZOH plasma levels. ITZOH was up to 3 times higher in the plasma of patients (p < 0.001), which can contribute to the antifungal treatment and suggests an important participation of this metabolite in therapy. Lastly, we evaluated three patients who underwent high dose 600 mg/day ITZ with a significant increase of ITZOH (p= 0.0148) plasma levels. In summary, our data 1) confirmed F. pedrosoi as the main agent of CBM in Pará; 2) did not show correlation between the lesions, species and ITZ sensitivity; 3) Showed that ITZ, VCZ and PCZ are the drugs with lower CIM, wherein PCZ had the lowest MFC; 4) correlated the metabolite ITZOH with the clinical evolution of patients and; 5) indicated that the use of high doses of up to 600 mg/day ITZ can be used in patients who do not respond to lower therapeutic doses.
Acesso aberto (Open Access)
Avaliação in vitro da atividade antifúngica da Malpighia glabra linn. em agentes da cromoblastomicose
(Universidade Federal do Pará, 2017-08-29) CRUZ, Naila Ferreira da; SILVA, Moises Batista da; http://lattes.cnpq.br/5525661855611118; SALGADO, Claudio Guedes; http://lattes.cnpq.br/2310734509396125
Chromoblastomycosis (CBM) is a mycosis by implantation, chronic, with a cosmopolitan distribution, caused by melanized fungi. After transcutaneous implantation, the propagules of the CBM agents present a unique cellular and morphological plasticity. The cellular differentiation results in the appearance of muriform cells. The treatment of this mycosis is a challenge due to the absence of a standard antifungal, resulting in several therapy methods (physical and pharmacological), used isolated or associated, with little clinical cure success. Amazon region has a vast biodiversity of vegetables that need to be better characterized as to their chemical composition and applicability in the treatment of different diseases. In this variety of plants, the fruit of Malpighia glabra Linn. (acerola) has a high content of vitamin C, also presenting vitamins of the complex B, A, anthocyanins and flavonoids, which stand out for having different biological and therapeutic actions already demonstrated both in vitro and in vivo. A major interest is currently focused on the biological activities of quercetin, belonging to the class of flavonoids, as it exerts multiple pharmacological activities, presenting unique biological properties that can improve mental and/or physical performance and reduce the risk of different infections. This study aims to evaluate the in vitro antifungal activity of M. glabra extract in conidia and muriform cells of Fonsecaea spp. The extract of M. glabra presented antifungal activity in both, conidia and muriform cells, of the evaluated strains. Much interest is now centered on biological activities of the flavonoid quercetin. In addition to the crude extract, the conidia of different strains were also sensitive to different dilutions of the extract. The minimum inhibitory concentration (MIC) and the minimum fungicidal concentration (MFC) were evaluated. The geometric mean of the quercetin MIC for the conidia was 4.75 μg/mL and the geometric mean of the CFM was 11.31 μg/mL. Further study is needed so that M. glabra or quercetin, isolated or associated with another isolated component of the extract, may be used for the treatment of CBM in the future.
Acesso aberto (Open Access)
Clinical, epidemiological and mycological report on 65 patients from the Eastern Amazon region with chromoblastomycosis
(2012-08) PIRES, Carla Andréa Avelar; XAVIER, Marília Brasil; QUARESMA, Juarez Antônio Simões; MACEDO, Geraldo Mariano Moraes de; SOUSA, Bruna Ranyelle de Marinho; BRITO, Arival Cardoso de
BACKGROUND: Chromoblastomycosis is a chronic fungal infection caused by fungi from the Dematiaceae family. According to several studies, Fonsecaea pedrosoi is the most common of these fungi. The infection is more common in tropical countries, with the Brazilian state of Pará having one of the largest infected populations worldwide. The disease is difficult to treat and recurrences are common. OBJECTIVES: To describe the epidemiological and mycological aspects of cases of chromoblastomycosis and its clinical forms in the state of Pará, Brazil. METHODS: Mycological exams (direct mycological examinations, culture and microculture) were performed and a clinical/epidemiological evaluation was made of 65 patients receiving care at the Dermatology Department of the Federal University of Pará between 2000 and 2007. The clinical classification proposed by Carrión in 1950 was used in this study. RESULTS: The majority of the patients were male (93.8%), agricultural workers (89.2%) of 45 to 55 years of age, and the majority of lesions (55.4%) were of the verrucous type, located principally on the lower limbs (81.5%). In the majority of the cases investigated (61.5%), the infection had been present for a long time, with a mean duration of 11 years. Direct mycological examination was performed in 86.2% of the patients (n=56). Of these, 96.4% (n=54) tested positive. Culture and microculture were performed in vitro in 47 cases of those that tested positive at direct microscopy, results showing Fonsecaea pedrosoi to be the only agent present in this sample. CONCLUSION: This study highlighted the extent to which chromoblastomycosis still affects the quality of life of the local population, principally individuals working in agriculture. This is a chronic disease for which there is no effective treatment. The importance of continuing to investigate this disease should be emphasized, as further studies may lead to new clinical or epidemiological findings.
Acesso aberto (Open Access)
Determinação das concentrações plasmáticas e teciduais de itraconazol em pacientes com cromoblastomicose
(Universidade Federal do Pará, 2008-09-01) GRISÓLIA, Daniella Paternostro de Araújo; VIEIRA, José Luiz Fernandes; http://lattes.cnpq.br/2739079559531098
Chromoblastomycosis is a subcutaneous mycosis caused by deployment transcutaneous of several species of dematiaceous fungi, that is, melanized fungi. Considering the incidence of this disease in the state of Pará and the resulting morbidity of patients affected, with economic and social repercussions, it was made to the optimization of therapeutic schemes adopted, to the best knowledge of the relation dose x response. The itraconazole is one of the few drugs available for treatment, which has marked variability kinetic intra and inter individual, which compromises the establishment of the relation dose and response, as well as tissue and plasma concentrations achieved. In this sense, this work aimed at validation of analytical methodology by High Performance Liquid Chromatography and subsequent determination of itraconazole in samples of plasma and tissue in 20 patients with chromoblastomycosis, assisted in the laboratory of dermatoimmunology Dr. Marcello Candia, Marituba, Pará, who used the drug in doses of 200mg/day and 400mg/day. The technique employed was validated and proved adequate results in accordance with applicable law. Concentrations of plasma and tissue of itraconazole in the dose of 200mg/day were 121.3 87.9 ng/mL and 5.36 5.9 μg/g. The average plasma concentration of itraconazole in patients using 400mg/day was 290 234 ng/mL, and the plasma and tissue mean concentrations of itraconazole in patients who showed no clinical favourable, at doses of 200mg, making it necessary to increase to 400mg were 217 216 and 304 173 ng/mL; 14.87 12.94 e 21.80 6.62 μg/g. The average of relation between tissue and plasma concentrations in patients who had positive developments in clinical in the doses of 200mg/day was 44.29 67.12 and those that did not show positive developments in clinical in the doses of 200mg/day, making necessary to increase to 400mg/day were 68.52 59.90 and 71.71 38.26 respectively.
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Indução de escleróticas in vitro e análise da resposta imune dos pacientes de cromoblastomicose em tratamento com itraconazol
(Universidade Federal do Pará, 2009-06-26) SILVA, Moisés Batista da; SALGADO, Claudio Guedes; http://lattes.cnpq.br/2310734509396125
Chromoblastomycosis (CBM) is a chronic fungal disease witch affects the skin, characterized for slowing development of polymorphic skin, that present infiltrated inflammatory granulomatous in the presence of sclerotics cells, characteristic of this illness. One of the objectives of this study was to evaluate the induction of scleroticts cells for natural mediums, with biomasses of Bactris gasipaes and Theobroma grandiflorum, whose respective species had induced in vitro similar sclerotics cells to those found in tissue of patients, in 10 and 2 days, respectively, what it made possible the production of a powder medium inductor, already donated to other groups that study the CBM. Another objective was to evaluate the histopathology of the CBM in the patients, before and during the use of itraconazole (ITZ). For this, the technique of ELISA for the cytokines was used TNF-α, circulating IL-4 and IL-10, and the immunohistochemestry of biópsias in different times of treatment - that it allowed to analyze the quantitative and qualitative alterations of the cellular types during 12 months of the treatment with ITZ in the 200 dose of mg/dia - with antibodies anti- CD20, anti-CD8 and anti-CD68. How much the cytokines, the circulating IL-10 did not show significant change, while IL-4 and TNF-α had presented an increase of the levels throughout 12 months of treatment. In relation to the immunophenotyping, it had a significant reduction in the inflammatory process and the cellular infiltrated during 3 and 6 months of the treatment, whereas only to the 12 months had the significant regression of the number of sclerotics cells. The immunophenotyping disclosed that the macrophages are mainly located in the areas central areas of granuloma, whereas cells TCD8+ are in the periphery and cells TCD20+, which were found throughout the tissues, with a significant increase after 6 months of the treatment, returning to the initial levels after one year. The cytotoxic macrophages and lymphocytes were having presented a significant increase after 12 months of treatment with ITZ. These results demonstrate that the formation of granuloma in the CBM is similar to those observed in other granulomatous infectious disease, and that the presence of IL-4 and IL-10 can be related with the persistence of fungi in the injuries and with the difficulty of cure observed in these patients.
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Isolation of Fonsecaea pedrosoi from thorns of Mimosa pudica, a probable natural source of chromoblastomycosis
(2004-02) SALGADO, Claudio Guedes; SILVA, Jorge Pereira da; DINIZ JUNIOR, José Antônio Picanço; SILVA, Moisés Batista da; COSTA, Patricia Fagundes da; TEIXEIRA, Cláudio Eduardo Corrêa; SALGADO, Ubirajara Imbiriba
We report the isolation of Fonsecaea pedrosoi from thorns of the plant Mimosa pudica L. at the place of infection identified by one of our patients. Clinical diagnosis of chromoblastomycosis was established by direct microscopic examination and cultures from the patient's lesion. The same species was isolated from the patient and from the plant. Scanning electron microscopy of the surface of the thorns showed the characteristic conidial arrangement of F. pedrosoi. These data indicate that M. pudica could be a natural source of infection for the fungus F. pedrosoi.