Role of iron in the nitric oxide-mediated fungicidal mechanism of IFN-gamma-activated murine macrophages against Paracoccidioides brasiliensis conidia

Iron is an essential growth element of virtually all microorganisms and its restriction is one of the mechanisms used by macrophages to control microbial multiplication. Paracoccidioides brasiliensis, the agent of paracoccidioidomycosis, an important systemic mycosis in Latin America, is inhibited in its conidia-to-yeast conversion in the absence of iron. We studied the participation of iron in the nitric oxide (NO)-mediated fungicidal mechanism against conidia. Peritoneal murine macrophages activated with 50U/mL of IFN-gamma or treated with 35 µM Deferoxamine (DEX) and infected with P. brasiliensis conidia, were co-cultured and incubated for 96 h in the presence of different concentrations of holotransferrin (HOLO) and FeS0(4). The supernatants were withdrawn in order to assess NO2 production by the Griess method. The monolayers were fixed, stained and observed microscopically. The percentage of the conidia-to-yeast transition was estimated by counting 200 intracellular propagules. IFN-gamma-activated or DEX-treated Mthetas presented marked inhibition of the conidia-to-yeast conversion (19 and 56%, respectively) in comparison with non-activated or untreated Mthetas (80%). IFN-gamma-activated macrophages produced high NO levels in comparison with the controls. Additionally, when the activated or treated-macrophages were supplemented with iron donors (HOLO or FeSO4), the inhibitory action was reversed, although NO production remained intact. These results suggest that the NO-mediated fungicidal mechanism exerted by IFN-gamma-activated macrophages against P. brasiliensis conidia, is dependent of an iron interaction.

On the unknown ecological niche of Paracoccidioides brasiliensis: our hypothesis of 1989: present status and perspectives

In 1989, CONTI DÍAZ & RILLA published a hypothesis concerning the as yet unknown ecological niche of Paracoccidioides brasiliensis. The hypothesis proposed a highly efficient paracoccidioidal ecological strategy based on an important natural reservoir of the parasite, probably in heterothermic animals from fresh water environments. Further, the hypothesis proposed, a transient and variable residence in the soil with a wide aleuriospore dispersion throughout the environment together with an elevated capability of infecting humans, and domestic and wild animals. This paper analyzes scientific publications from the IX International Meeting on Paracoccidioidomycosis held in Águas de Lindoia, São Paulo, Brazil from 2-5 October 2005, providing a comparative study among this articles and with other recently published papers and the hypothesis' postulates. Since various findings and observations appear to agree with the postulates, the pursuit of novel, specific research projects in the supposed reservoirs is recommended partially or fully to confirm the hypothesis using classical laboratorial methods and modern molecular biology techniques.

Characterization of green zero-valent iron nanoparticles produced with tree leaf extracts

In the last decades nanotechnology has become increasingly important because it offers indisputable advantages to almost every area of expertise, including environmental remediation. In this area the synthesis of highly reactive nanomaterials (e.g. zero-valent iron nanoparticles, nZVI) is gaining the attention of the scientific community, service providers and other stakeholders. The synthesis of nZVI by the recently developed green bottom-up method is extremely promising. However, the lack of information about the characteristics of the synthetized particles hinders a wider and more extensive application. This work aims to evaluate the characteristics of nZVI synthesized through the green method using leaves from different trees. Considering the requirements of a product for environmental remediation the following characteristics were studied: size, shape, reactivity and agglomeration tendency. The mulberry and pomegranate leaf extracts produced the smallest nZVIs (5–10 nm), the peach, pear and vine leaf extracts produced the most reactive nZVIs while the ones produced with passion fruit, medlar and cherry extracts did not settle at high nZVI concentrations (931 and 266 ppm). Considering all tests, the nZVIs obtained from medlar and vine leaf extracts are the ones that could present better performances in the environmental remediation. The information gathered in this paper will be useful to choose the most appropriate leaf extracts and operational conditions for the application of the green nZVIs in environmental remediation.

Lysozyme plays a dual role against the dimorphic fungus Paracoccidioides brasiliensis

In order to determine the role of lysozyme, an antimicrobial peptide belonging to the innate immune system, against the dimorphic fungus Paracoccidioides brasiliensis, co-cultures of the MH-S murine alveolar macrophages cell line with P. brasiliensis conidia were done; assays to evaluate the effect of physiological and inflammatory concentrations of lysozyme directly on the fungus life cycle were also undertaken. We observed that TNF-α-activated macrophages significantly inhibited the conidia to yeast transition (p = 0.0043) and exerted an important fungicidal effect (p = 0.0044), killing 27% more fungal propagules in comparison with controls. Nonetheless, after adding a selective inhibitor of lysozyme, the fungicidal effect was reverted. When P. brasiliensis propagules were exposed directly to different concentrations of lysozyme, a dual effect was observed. Physiologic concentrations of the enzyme facilitated the conidia-to-yeast transition process (p < 0.05). On the contrary, inflammatory concentrations impaired the normal temperature-dependant fungal transition (p < 0.0001). When yeast cells were exposed to lysozyme, irrespective of concentration, the multiple-budding ability was badly impaired (p < 0.0001). In addition, ultra-structural changes such as subcellular degradation, fusion of lipid vacuoles, lamellar structures and interruption of the fibrilar layer were observed in lysozyme exposed conidia. These results suggest that lysozyme appears to exert a dual role as part of the anti-P. brasiliensis defense mechanisms.

Identification of Paracoccidioides brasiliensis in adrenal glands biopsies of two patients with paracoccidioidomycosis and adrenal insufficiency

The authors report two cases of adrenal insufficiency secondary to infiltration of the adrenal glands by Paracoccidioides brasiliensis. The first patient had been treated for a chronic multifocal form of paracoccidiodomycosis 11 years ago. The diagnosis of the mycosis was done simultaneous with that of the adrenal insufficiency in the second patient. In both patients the diagnosis was done by direct visualization of fungus in adrenal biopsies. They were treated with hormonal supplements and itraconazol by 12 and six months, without relapses during the follow-up period.

DOSE RESPONSE EFFECT OF Paracoccidioides brasiliensis IN AN EXPERIMENTAL MODEL OF ARTHRITIS

Paracoccidioidomycosis (PCM) is caused by the dimorphic fungus Paracoccidioides brasiliensis (Pb) and corresponds to prevalent systemic mycosis in Latin America. The aim of the present work was to evaluate the dose response effect of the fungal yeast phase for the standardization of an experimental model of septic arthritis. The experiments were performed with groups of 14 rats that received doses of 103, 104 or 105 P. brasiliensis (Pb18) cells. The fungi were injected in 50 µL of phosphate-buffered saline (PBS) directly into the knee joints of the animals. The following parameters were analyzed in this work: the formation of swelling in knees infused with yeast cells and the radiological and anatomopathological alterations, besides antibody titer by ELISA. After 15 days of infection, signs of inflammation were evident. At 45 days, some features of damage and necrosis were observed in the articular cartilage. The systemic dissemination of the fungus was observed in 11% of the inoculated animals, and it was concluded that the experimental model is able to mimic articular PCM in humans and that the dose of 105 yeast cells can be used as standard in this model.

THE POWER OF THE SMALL: THE EXAMPLE OF Paracoccidioides brasiliensis CONIDIA

SUMMARYResearch on Paracoccidioides brasiliensis has centered in the yeast cell probably because of the lack of distinctive features in the mycelium. In 1942 and for the first time, lateral conidia were noticed in the fungus' hyphae. Later on, Brazilian, Venezuelan and Argentinean researchers described "aleurias" when the fungus was grown in natural substrates. In 1970 authors became interested in the conidia and were able to obtain them in large numbers and treat them as individual units. Their shape and size were defined and the presence of all the elements of a competent eukaryotic cell were demonstrated. Conidia exhibited thermal dimorphism and, additionally, when given intranasally to BALB/c male mice, they converted into yeasts in the lungs and produce progressive pulmonary lesions with further dissemination to other organs. Studies on the phagocyte-conidia interaction were revealing and showed that these versatile structures allow a better understanding of the host- P. brasiliensisinteractions.

Paracoccidioides brasiliensis AND Paracoccidioides lutzii, A SECRET LOVE AFFAIR

SUMMARYTo commemorate Prof. Carlos da Silva Lacaz's centennial anniversary, the authors have written a brief account of a few, out of hundreds, biological, ecological, molecular and phylogenetic studies that led to the arrival of Paracoccidioides lutzii, hidden for more than a century within Paracoccidioides brasiliensis. Lacaz's permanent interest in this fungus, and particularly his conviction on the benefits that research on paracoccidioidomycosis would bring to patients, were pivotal in the development of the field.

Ciclo Vital de Schistosoma mansoni através do Holochilus brasiliensis (Desmarest, 1818) em ambiente semi-natural (Trematoda, Shistosomatidae; Rodentia, Cricetidae)

Junto ao Lago da Pampulha, Belo Horizonte, MG, foram capturados (julho/72-novembro/73) 28 exemplares de Holochilus brasiliensis, dos quais 11 (39,3%) eliminavam nas fezes ovos viáveis de S. mansoni. Miracídios da cepa mencionada ("H") infectaram Biomphalaria glabrata e as cercárias obtidas também infectaram camundongos albinos, recuperando-se, ao final do experimento, 35,3% de vermes adultos. Por outro lado, cercárias de cepa humana ("LE") de S. mansoni infectaram sete exemplares de H. brasiliensis, nascidos em laboratório, recuperando-se no fim de 60 dias, 30,5% de vermes adultos. Estudos anatomapatológicos de H. brasiliensis demonstraram infecção generalizada, encontrando-se granuloma no esôfago, estômago, intestino (delgado e grosso), fígado, baço, pâncreas e linfonodos abdominais. Espessamentos fibrosos da íntima da veia porta, granulomas em espaços porta e fibrose incipiente dos espaços porta e interlobular foram lesões decorrentes da presença de ovos de S. mansoni encontrados no fígado. Em ambiente semi-natural, foi possível fechar o ciclo do S. mansoni sem direta participação humana, utilizando-se B. glabrata experimentalmente infectadas com trematódeos da cepa "LE", H. brasiliensis nascidos em laboratório e B. glabrata nascida no ambiente semi-natural estabelecido. Verificou-se que ambas as cepas ("H" e "LE") comportaram-se de maneira análoga, não sendo verificadas, também, diferenças morfológicas entre os ovos e vermes adultos de ambas. As observações, realizadas no campo e no laboratório demonstraram que o Holochilus brasiliensis é bom hospedeiro de Schistosoma mansoni. Assim, em determinadas áreas e sob certas condições ecológicas, o cricetídeo em questão poderá, efetivamente, integrar-se ao ciclo do trematódeo na natureza, independente ou paralelamente à presença do homem. Assinala-se, finalmente, que o presente trabalho relata o segundo fechamento do ciclo biológico de S. mansoni em condições ditas semi-naturais. Os primeiros estudos, entretanto, de Antunes, Milward de Andrade, Katz & Coelho4,,em 1971 e de Antunes5, em 1971 foram feitos utilizando-se o Nectomys s. squamipes.

The palm tree Copernicia cerifera (carnaúba) as an ecotope of Rhodnius nasutus in rural areas of the State of Piauí Northeastern Brazil

The C. cerifera palm tree (carnaúba) is widely distributed in the Northeastem Brazil, including the State of Piauí. This investigation revealed that R. nasutus is the ortly triatomine species captured on that palm tree, in five different localities. 78% of palm trees were infested with triatomines, and 4.0% were infected with flagellates morphologically and biologically indistinguishable from Trypanosoma cruzi. Birds, rodents and marsupials were found as major blood meai sources for R. nasutus.

Utilização do picro-sirius para corar o Paracoccidioides brasiliensis

A solução do picro-sirius foi usada para corar o Paracoccidioides brasiliensis em tecidos humanos fixados em formol. À luz comum, o fungo mostra uma faixa periférica avermelhada e, à luz polarizada, apresenta um ou mais anéis birrefringentes esverdeados. A imersão prévia em solução de hidróxido de sódio a 1%, por alguns minutos, melhora a coloração, sendo o fungo visualizado com mais nitidez à luz polarizada. Por outro lado, a diferenciação dos cortes corados, com picro-sirius em hidróxido de sódio a 0,5%, elimina a birrefringência do colágeno mais rapidamente que a do Paracoccidioides.

Influence of light conditions on adult mortality and egg-laying of Triatoma brasiliensis neiva, 1911 (Hemiptera: Reduviidae: Triatominae)

We compared, for Triatoma brasiliensis, the egg-laying process and the mortality of adults under conditions of almost permanent darkness and with normal laboratory luminosity. Mortality did not differ between groups. The egg-laying per vial and per female was significantly greater in the group of normal luminosity. We consider that it is not recommendable to keep the adults of this species under complete darkness. Other biological aspects should be analysed in relation to luminosity.

Avaliaçao do efeito do extrato de casca de cajueiro-branco (Anacardium occidentale L.) Sobre A infecção por Leishmania (Viannia) brasiliensis

Este trabalho apresenta uma avaliação do efeito terapêutico do extrato hidroalcólico da casca de Anacardium occidentale L sobre a Leishmania (Viannia) brasiliensis. No modelo in vitro o extrato vegetal mostrou-se ativo contra promastigotas do parasita, contrastando com o modelo in vivo, onde não se observou qualquer atividade curativa.

Avaliação do cetoconazol em camundongos inoculados com Paracoccidioides brasiliensis pela histopatologia de fígado e baço e pela intradermorreação de paracoccidioidina

Camundongos machos albinos, foram inoculados por via intravenosa com 0,5xl07 células leveduriform.es viáveis de Paracoccidioides brasiliensis da amostra rotulada como Pb 2052 e tratados diariamente com cetoconazol nas dosagens de 50 e 100mg/kg durante 50 dias e em seguida sacrificados. Observou-se o número de parasitas .formação de granulomas no fígado e baço e reação de hipersensibilidade retardada usando-se antígeno de P. brasiliensís preparada segundo Fava Neto, inoculado na pata direita do animal. Verificou-se que: 1. cetoconazol diminuiu acentuadamente o número de parasitas encontrados no fígado e baço; 2. não havia diferença entre o número de granulomas formados no baço e fígado de animais tratados e não tratados; 3. a intensidade dos testes cutâneos tardios foram similares em todos os animais.