Abstract This study aimed to evaluate the in vitro antifungal activity of terpinen-4-ol, tyrosol, and β-lapachone against strains of Coccidioides posadasii in filamentous phase (n = 22) and Histoplasma capsulatum in both filamentous (n = 40) and yeast phases (n = 13), using the broth dilution methods as described by the Clinical and Laboratory Standards Institute, to determine the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of these compounds. The mechanisms of action of these compounds were also investigated by analyzing their effect on cell membrane permeability and ergosterol synthesis. The MIC and MFCf these compounds against C. posadasii, mycelial H. capsulatum, and yeast-like H. capsulatum, were in the following ranges: 350-5720 µg/mL, 20-2860 µg/mL, and 40-1420 µg/mL, respectively for terpinen-4-ol; 250-4000 µg/mL, 30-2000 µg/mL, and 10-1000 µg/mL, respectively, for tyrosol; and 0.48-7.8 µg/mL, 0.25-16 µg/mL, and 0.125-4 µg/mL, respectively for β-lapachone. These compounds showed a decrease in MIC when the samples were subjected to osmotic stress, suggesting that the compounds acted on the fungal membrane. All the compounds were able to reduce the ergosterol content of the fungal strains. Finally, tyrosol was able to cause a leakage of intracellular molecules.

ABSTRACTThe antifungal activity of some statins against different fungal species has been reported. Thus, at the first moment, the in vitro antifungal activity of simvastatin, atorvastatin and pravastatin was tested againstCandida spp. and Cryptococcus spp. Then, in a second approach, considering that the best results were obtained for simvastatin, this drug was evaluated in combination with antifungal drugs against planktonic growth and tested against biofilms ofCandida spp. and Cryptococcus spp. Drug susceptibility testing was performed using the microdilution broth method, as described by the Clinical and Laboratory Standards Institute. The interaction between simvastatin and antifungals against planktonic cells was analyzed by calculating the fractional inhibitory concentration index. Regarding biofilm susceptibility, simvastatin was tested against growing biofilm and mature biofilm of one strain of each tested yeast species. Simvastatin showed inhibitory effect against Candida spp. andCryptococcus spp. with minimum inhibitory concentration values ranging from 15.6 to 1000 mg L-1 and from 62.5 to 1000 mg L-1, respectively. The combination of simvastatin with itraconazole and fluconazole showed synergism against Candidaspp. and Cryptococcus spp., while the combination of simvastatin with amphotericin B was synergistic only againstCryptococcus spp. Concerning the biofilm assays, simvastatin was able to inhibit both growing biofilm and mature biofilm ofCandida spp. and Cryptococcus spp. The present study showed that simvastatin inhibits planktonic cells and biofilms ofCandida and Cryptococcus species.

Este estudo teve como objetivo avaliar a atividade antifúngica de extratos de M. oleifera frente a fungos isolados de camarões, cultivados em água doce, e testar a toxicidade dos extratos em larvas de Macrobrachium amazonicum. Os extratos etanólicos de vagens, sementes, folhas, caules e flores e o extrato clorofórmico de flores de M. oleifera foram testados contra 14 cepas de Candida spp. e 10 cepas de Hortaea werneckii isolados da água de cultivo e do trato digestório de M. amazonicum. A atividade antifúngica foi determinada por microdiluição, com base nos documentos M27-A3 e M38-A2 do CLSI. A toxicidade foi avaliada por exposição das larvas de M. amazonicum a concentrações entre 10-1000 mg mL-1 dos extratos, realizando contagem de larvas mortas (CL50), após 24 horas. Os melhores resultados foram verificados com o extrato clorofórmico de flores, agindo frente a todas as cepas testadas, com concentrações inibitórias mínimas variando entre 0,019-2,5 mg mL-1. O extrato etanólico de folhas, flores e sementes agiu ante 22/24, 21/24 e 20/24 cepas, respectivamente. O extrato de vagens foi eficaz contra cepas de Candida spp. (14/24) e o extrato de caule apenas contra quatro cepas de H. werneckii (4/24). Os extratos de sementes, flores (fração clorofórmica), caules e folhas apresentaram baixa ou nenhuma toxicidade, enquanto que extratos de vagens e flores (fração etanólica) apresentaram toxicidade moderada. Assim, observou-se atividade antifúngica dos extratos em Candida spp . e H. werneckii com uma ampla margem de segurança para as larvas de M. amazonicum, demonstrando ser promissor para o manejo sustentável dos efluentes do cultivo de M. amazonicum

This study aimed to evaluate the antifungal activity of M. oleifera extracts against fungi isolated from farmed prawns and test the toxicity of the extracts on larvae of Macrobrachium amazonicum. The ethanol extracts of pods, seeds, leaves, stems and flowers and chloroform extract of flowers of M. oleifera were tested against 14 strains of Candida spp. and 10 strains of Hortaea werneckii isolated from farming water and the digestive tract of M. amazonicum. Antifungal activity was determined by microdilution, based on the M27-A3 and M38-A2 CLSI documents. Toxicity was evaluated by exposing larvae of M. amazonicum at concentrations between 10-1000mg mL-1, counting dead larvae (CL50) after 24 hours. The best results were verified with the chloroform extract of flowers, acting against all tested strains, with MICs ranging from 0.019 to 2.5 mg mL-1. Ethanol extracts of leaves, flowers and seeds acted against 22/24, 21/24 and 20/24 strains, respectively. The extract of pods was only effective against strains of Candida spp. (14/24) and extract of stem only against four strains of H. werneckii (4/24). Extracts of seeds, flowers (chloroform fraction), stems and leaves showed low or no toxicity, whereas extracts of pods and flowers (ethanol fraction) showed moderate toxicity. Thus, the antifungal activity of these extracts agaisnt Candida spp. and H. werneckii was observed, a wide margin of safety for larvae of M. amazonicum, demonstrating to be promising for the sustainable management of effluents from M. amazonicum farming

Este estudo teve como objetivo avaliar o papel do camarão Macrobrachium amazonicum como carreador de Candida spp. do ambiente aquático, por meio da análise da correlação entre Candida spp. isoladas desse camarão e do seu ambiente (água de superfície e sedimento) através das técnicas de M13-PCR fingerprinting e RAPD-PCR. Para tanto, 27 cepas de Candida spp. foram avaliadas. Essas cepas foram recuperadas a partir do trato gastrointestinal de M. amazonicum adultos (7/27), oriundos da lagoa do Catú, Ceará, Brasil, e do meio aquático (águas superficiais e sedimentos) desse lago (20/27). A comparação molecular entre as cepas de camarões e o ambiente aquático foi conduzida por M13-PCR fingerprinting e RAPD-PCR, utilizando os iniciadores M13 e OPQ16, respectivamente. A análise molecular revelou semelhanças entre os padrões de bandas de oito isolados de Candida com o iniciador M13 e 11 isolados com o primer OPQ16, indicando que elas estão presentes no trato digestivo de M. amazonicum e no ambiente aquático, onde esses camarões habitam. Portanto, essa espécie de camarão pode ser usada como sentinela para monitoramento ambiental através da recuperação de Candida spp. do ambiente aquático, a partir do seu trato gastrointestinal

This study aimed to evaluate the role of the Amazon River prawn, Macrobrachium amazonicum, as carrier of Candida spp., by analyzing the correlation between Candida spp. from these prawns and their environment (surface water and sediment), through M13-PCR fingerprinting and RAPD-PCR. For this purpose, 27 strains of Candida spp. were evaluated. These strains were recovered from the gastrointestinal tract of adult M. amazonicum (7/27) from Catú Lake, Ceará State, Brazil and from the aquatic environment (surface water and sediment) of this lake (20/27). Molecular comparison between the strains from prawns and the aquatic environment was conducted by M13-PCR fingerprinting and RAPD-PCR, utilizing the primers M13 and OPQ16, respectively. The molecular analysis revealed similarities between the band patterns of eight Candida isolates with the primer M13 and 11 isolates with the primer OPQ16, indicating that the same strains are present in the digestive tract of M. amazonicum and in the aquatic environment where these prawns inhabit. Therefore, these prawns can be used as sentinels for environmental monitoring through the recovery of Candida spp. from the aquatic environment in their gastrointestinal tract

Soroprevalence for Hepatitis C virus is reported as 2.12% in Northern Brazil, with about 50% of the patients exhibiting a sustained virological response (SVR). Aiming to associate polymorphisms in Killer Cell Immunoglobulin-like Receptors (KIR) with chronic hepatitis C and therapy responses we investigated 125 chronic patients and 345 controls. Additionally, 48 ancestry markers were genotyped to control for population stratification. The frequency of the KIR2DL2 and KIR2DL2+HLA-C Asp80 gene and ligand was higher in chronic infected patients than in controls (p < 0.0009, OR = 3.4; p = 0.001, OR = 3.45). In fact, KIR2DL3 is a weaker inhibitor of NK activity than KIR2DL2, which could explain the association of KIR2DL2 with chronic infection. Moreover, KIR2DS2 and KIR2DS2+HLA-C Asp80 (p < 0.0001, OR = 2.51; p = 0.0084, OR = 2.62) and KIR2DS3 (p < 0.0001; OR = 2.57) were associated with chronic infection, independently from KIR2DL2. No differences in ancestry composition were observed between control and patients, even with respect to therapy response groups. The allelic profile KIR2DL2/KIR2DS2/KIR2DS3 was associated with the chronic hepatitis C (p < 0.0001; OR = 3). Furthermore, the patients also showed a higher mean number of activating genes and a lower frequency of the homozygous AA profile, which is likely secondary to the association with non-AA and/or activating genes. In addition, the KIR2DS5 allele was associated with SVR (p = 0.0261; OR = 0.184).The ancestry analysis of samples ruled out any effects of population substructuring and did not evidence interethnic differences in therapy response, as suggested in previous studies.

A histoplasmose é uma doença fúngica causada pela inalação de esporos de Histoplasma capsulatum. A maioria dos indivíduos normais não apresenta doença após pequena inalação, porém exposições mais prolongadas podem levar ao desenvolvimento de infecção pulmonar aguda, crônica ou disseminada. Nos pacientes imunocomprometidos a infecção é disseminada e grave. Relatamos o caso de um paciente de treze anos, imunocompetente, com febre, tosse seca e dispnéia progressiva havia dois meses. O radiograma e a tomografia computadorizada de tórax evidenciavam infiltrado intersticial com micronódulos difusos. O paciente relatava contato intenso com pássaros em sua residência. Foi submetido a biópsia pulmonar a céu aberto, que evidenciou Histoplasma capsulatum em tecido pulmonar. A cultura do fragmento da biópsia confirmou a presença de Histoplasma capsulatum sp. O paciente foi tratado com anfotericina-B por 28 dias, seguida de itraconazol por seis meses, com resolução do quadro.

Histoplasmosis is a fungal disease caused by inhalation of Histoplasma capsulatum fungus. The disease does not normally affect immunocompetent individuals after a single, transient inhalation exposure. However, longer exposure may cause chronic or disseminated acute pulmonary infection. In immunocompromised patients, the infection is disseminated and severe. We report the case of a 13-year-old immunocompetent patient, presenting with fever, cough and dyspnea for one month. The chest X-ray and computed tomography scan revealed interstitial infiltrate and diffuse micronodules. The patient reported having had close and prolonged contact with birds. He was submitted to an open lung biopsy and the tissue culture was positive for Histoplasma capsulatum sp. He was treated with amphotericin B for 28 days, followed by treatment with itraconazole for 6 months, and there was complete resolution of the disease.

INTRODUÇÃO: Os exames de cultura e o exame bacterioscópico pelo método de coloração de Gram (GRAM) do aspirado traqueal ainda são objeto de controvérsias com relação ao diagnóstico etiológico na pneumonia associada à ventilação mecânica (PAV). OBJETIVO: Avaliar a concordância entre os resultados do GRAM e da cultura quantitativa do aspirado traqueal e do lavado broncoalveolar nos pacientes com PAV. MÉTODO: Foram estudados de modo prospectivo os pacientes internados no período de outubro de 2001 a agosto de 2002, que estavam há mais de 48hs sob ventilação mecânica, e que apresentavam suspeita clínica de PAV. No momento da suspeita clínica foi realizado o aspirado traqueal seguido do lavado broncoalveolar. O diagnóstico de PAV foi confirmado com a suspeita clínica associada à cultura quantitativa do lavado broncoalveolar 10(4)ufc/ml. RESULTADOS: Dos 119 pacientes sob ventilação mecânica, 32 (26,8%) tiveram suspeita clínica de PAV, com confirmação diagnóstica em 25 (78%) deles. A comparação entre o GRAM do aspirado traqueal e a cultura do lavado broncoalveolar mostrou uma moderada concordância (coeficiente de Kappa de 0,56). Houve concordância entre a cultura quantitativa do aspirado traqueal e do lavado broncoalveolar em 22/25 (88%) e discordância em 3/25 (12%) casos (coeficiente de Kappa de 0,71). A sensibilidade e a especificidade do aspirado traqueal para o diagnóstico de PAV com o ponto de corte 10 6 ufc/ml foram de 71% e 72%, respectivamente. CONCLUSÃO: A combinação do GRAM com a cultura quantitativa do aspirado traqueal pode contribuir para a avaliação diagnóstica da PAV.

BACKGROUND: The value of endotracheal aspirate cultures and of the GRAM stain test for the etiologic diagnosis of ventilator-associated pneumonia (VAP) still remains controversial. OBJECTIVES: To assess the consistency between the results of quantitative cultures and GRAM stain tests of the endotracheal aspirates and the bronchoalveolar lavage in patients with VAP. METHOD: Between October/2001 and August/2002, we prospectively studied all patients under mechanical ventilation for more than 48 hours with clinical suspicion of VAP. At the time of clinical suspicion, an endotrachealaspirate followed by a bronchoalveolar lavage was performed. Diagnosis of VAP was defined when clinical suspicion was confirmed by a positive quantitative culture (>104cfu/ml) in the bronchoalveolar lavage. RESULTS: Among 119 patients, 32 (26.8%) were suspected of having VAP, with confirmation in 25 (78%). Comparison between GRAM test of endotracheal aspirates and culture results of the bronchoalveolar lavage showed a moderate consistency (Kappa coefficient: 0.56). The was consistency between quantitative cultures of endotracheal aspirate and bronoalveolar lavage in 22 (88%) at the patients and no consistency in 3 (12%). (Kappa coefficient: 0.71). The sensitivity and specificity of the endotracheal aspirate culture with cut off of 106 cfu/m were 72% and 71%, respectively. CONCLUSION: The combination of GRAM stain test and quantitative cultures of the endotracheal aspirates may contribute to the diagnostic evaluation of VAP.