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LyeTx I, an antimicrobial peptide isolated from the venom of Lycosa erythrognatha, known as wolf spider, has been synthesised and its structural profile studied by using the CD and NMR techniques. LyeTx I has shown to be active against bacteria (Escherichia coli and Staphylococcus aureus) and fungi (Candida krusei and Cryptococcus neoformans) and able to alter the permeabilisation of L: -alpha-phosphatidylcholine-liposomes (POPC) in a dose-dependent manner. In POPC containing cholesterol or ergosterol, permeabilisation has either decreased about five times or remained unchanged, respectively. These results, along with the observed low haemolytic activity, indicated that antimicrobial membranes, rather than vertebrate membranes seem to be the preferential targets. However, the complexity of biological membranes compared to liposomes must be taken in account. Besides, other membrane components, such as proteins and even specific lipids, cannot be discarded to be important to the preferential action of the LyeTx I to the tested microorganisms. The secondary structure of LyeTx I shows a small random-coil region at the N-terminus followed by an alpha-helix that reached the amidated C-terminus, which might favour the peptide-membrane interaction. The high activity against bacteria together with the moderate activity against fungi and the low haemolytic activity have indicated LyeTx I as a good prototype for developing new antibiotic peptides.

Photodynamic inhibition was more efficient in promoting cell death than the antifungal cyclopiroxolamine against T. rubrum. ROS, ONOO· and NO· were important in the fungicidal activity of aPI. A suggested mechanism for this activity is that TBO is excited by LED light (630 nm), reacts with biomolecules and increases the availability of transition electrons and substrates for nitric oxide synthase, thereby increasing the oxidative and nitrosative bursts in the fungal cell.

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Cryptococus gattii is an emergent primary human pathogen that causes meningismus, papilledema, high intracranial pressure and focal involvement of the central nervous system in immunocompetent hosts. Prolonged antifungal therapy is the conventional treatment, but it is highly toxic, selects for resistant strains, contributes to therapy failure and has a poor prognosis. Photodynamic inactivation (PDI) offers a promising possibility for the alternative treatment of cryptococcosis. The aim of this study was to test the effectiveness of toluidine blue O (TBO) and light-emitting diode (LED) against C. gattii strains with distinct susceptibility profile to antifungal drugs (amphotericin B: 0.015-1.0 μg mL(-1); itraconazole: 0.015-2 μg mL(-1); fluconazole: 4-64 μg mL(-1)). Using 25 μM (6.76 μg mL(-1)) TBO and LED energy density of 54 J cm(-2) these fungal isolates presented variable susceptibility to PDI. The production of reactive oxygen species (ROS)/peroxynitrite was determined, and the catalase and peroxidase activities were measured. After PDI, high amounts of ROS/peroxynitrite are produced and higher catalase and peroxidase activities could be correlated with a lower susceptibility of C. gattii isolates to PDI. These results indicate that PDI could be an alternative to C. gattii growth inhibition, even of isolates less susceptible to classical antifungal drugs, also pointing to mechanisms related to their variable susceptibility behavior.

Forty-three clinical isolates of Sporothrix schenckii derived from humans and animals were evaluated in vitro for their susceptibility to amphotericin B, itraconazole, and terbinafine. MICs were determined by the method of micro dilution in liquid media, using protocols M27-A2 for the yeast form and M38-A for the mycelial form, both standardized by the Clinical Laboratory Standards Institute. In general, higher MICs were found for the mycelial form (intervals of up to two dilutions). In the case of amphotericin B, a significant difference in activity was observed, with higher values (p<0.05) found for the mycelial form. MICs for itraconazole and terbinafine were similar for both yeast and mycelial forms but slightly higher for mycelia. Although data presented here indicate different levels of susceptibility when both growth forms were compared, indicating an intrinsic difference between them, it is still difficult to draw a consensus as to which form correlates better with clinical findings. More studies are necessary to determine the criteria for in vitro tests that will lead to efficient therapeutic choices.

BACKGROUND -Trichophyton rubrum is the most common agent of superficial mycosis of the skin and nails causing long lasting infections and high recurrence rates. Current treatment drawbacks involve topical medications not being able to reach the nail bed at therapeutic concentrations, systemic antifungal drugs failing to eradicate the fungus before the nails are renewed, severe side effects and selection of resistant fungal isolates. Photodynamic therapy (PDT) has been a promising alternative to conventional treatments. OBJECTIVES: This study evaluated the in vitro effectiveness of toluidine blue O (TBO) irradiated by Light emitting diode (LED) in the reduction of T. rubrum viability. METHODS: The fungal inoculums' was prepared and exposed to different TBO concentrations and energy densities of Light emitting diode for evaluate the T. rubrum sensibility to PDT and production effect fungicidal after photodynamic treatment. In addition, the profiles of the area and volume of the irradiated fungal suspensions were also investigated. RESULTS: A small reduction, in vitro, of fungal cells was observed after exposition to 100 μM toluidine blue O irradiated by 18 J/cm 2 Light emitting diode. Fungicidal effect occurred after 25 μM toluidine blue O irradiation by Light emitting diode with energy density of 72 J/cm 2 . The analysis showed that the area and volume irradiated by the Light emitting diode were 52.2 mm² and 413.70 mm³, respectively. CONCLUSIONS: The results allowed to conclude that Photodynamic therapy using Light emitting diode under these experimental conditions is a possible alternative approach to inhibit in vitro T. rubrum and may be a promising new treatment for dermatophytosis caused by this fungus. Keywords: Antifungal agents; Onychomycosis; Photochemotherapy Resumo: FUNDAMENTOS -Trichophyton rubrum é o agente mais comum das micoses superficiais de pele e unhas causando infecções de longa duração e altas taxas de recidiva. As desvantagens do tratamento atual envolvem medicações tópicas as quais não são capazes de alcançar o leito ungueal em concentrações terapêuticas, antifúngicos sistêmicos que não erradicam o fungo antes das unhas serem renovadas, efeitos colaterais graves e seleção de isolados fúngicos resistentes. A terapia fotodinâmica tem sido uma alternativa promissora aos tratamentos convencionais. OBJETIVOS -Este estudo avaliou a eficácia, in vitro, de azul de orto-toluidina irradiado por diodo emissor de luz na redução da viabilidade de T. rubrum. MÉTODOS: O inóculo fúngico foi preparado e exposto a diferentes concentrações de azul de orto-toluidina e densidades de energia do diodo emissor de luz, para avaliar a sensibilidade de T. rubrum e o efeito fungicida, após terapia fotodinâmica. Além disso, os perfis da área e volume das suspensões fúngicas irradiados também foram investigados. RESULTADOS: Uma pequena redução, in vitro, de células fúngicas foi observada após a exposição a 100 mM azul de orto-toluidina irradiados por diodo emissor de luz a 18 J/cm 2 . Efeito fungicida ocorreu a...

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