Paracoccidioides brasilinsis-Induced Migration of Dendritic Cells and Subsequent T-Cell Activation in the Lung-Draining Lymph Nodes

Paracoccidioidomycosis is a mycotic disease caused by a dimorphic fungus, Paracoccidioides brasiliensis (Pb), that starts with inhalation of the fungus; thus, lung cells such as DC are part of the first line of defense against this microorganism. Migration of DC to the lymph nodes is the first step in initiating T cell responses. The mechanisms involved in resistance to Pb infection are poorly understood, but it is likely that DC play a pivotal role in the induction of effector T cells that control Pb infection. In this study, we showed that after Pb Infection, an important modification of lung DC receptor expression occurred. We observed an increased expression of CCR7 and CD103 on lung DC after infection, as well as MHC-II. After Pb infection, bone marrow-derived DC as well lung DC, migrate to lymph nodes. Migration of lung DC could represent an important mechanism of pathogenesis during PCM infection. In resume our data showed that Pb induced DC migration. Furthermore, we demonstrated that bone marrow-derived DC stimulated by Pb migrate to the lymph nodes and activate a T helper (Th) response. To the best of our knowledge, this is the first reported data showing that Pb induces migration of DC and activate a T helper (Th) response.

Dendritic Cells Transfected with scFv from Mab 7.B12 Mimicking Original Antigen gp43 Induces Protection against Experimental Paracoccidioidomycosis

Paracoccidioidomycosis (PCM), endemic in Latin America, is a progressive systemic mycosis caused by Paracoccidioides brasiliensis (P. brasiliensis), which primarily attacks lung tissue. Dendritic cells (DCs) are able to initiate a response in naive T cells, and they also participate in Th-cell education. Furthermore, these cells have been used for therapy in several disease models. Here we transfected DCs with a plasmid (pMAC/PS-scFv) encoding a single chain variable fragment (scFv) of an anti-Id antibody that is capable of mimicking gp43, the main antigenic component of P. brasiliensis. First, Balb/c mice were immunized subcutaneously with pMAC/PS-scFv and, after seven days, scFv protein was presented to the regional lymph nodes cells. Moreover, we showed that the DCs transfected with scFv were capable of efficiently activating proliferation of total lymph node cells and inducing a decrease in lung infection. Therefore, our results suggested that the use of scFv-transfected DCs may be a promising therapy in the paracoccidioidomycosis (PCM) model.

T Helper 1-Inducing Adjuvant Protects against Experimental Paracoccidioidomycosis

Immunostimulatory therapy is a promising approach to improving the treatment of systemic fungal infections such as paracoccidioidomycosis (PCM), whose drug therapy is usually prolonged and associated with toxic side effects and relapses. The current study was undertaken to determine if the injection of a T helper (Th) 1-stimulating adjuvant in P. brasiliensis infected mice could have a beneficial effect on the course of experimental PCM. For this purpose, mice were infected and treated with complete Freund's adjuvant (CFA), a well-established Th1 experimental inductor, or incomplete Freund's adjuvant (IFA - control group) on day 20 postinfection. Four weeks after treatment, the CFA-treated mice presented a mild infection in the lungs characterized by absence of epithelioid cell granulomas and yeast cells, whereas the control mice presented multiple sites of focal epithelioid granulomas with lymphomonocytic halos circumscribing a high number of viable and nonviable yeast cells. In addition, CFA administration induced a 2.4 log reduction (>99%) in the fungal burden when compared to the control group, and led to an improvement of immune response, reversing the immunosuppression observed in the control group. The immunotherapy with Th1-inducing adjuvant, approved to be used in humans, might be a valuable tool in the treatment of PCM and potentially useful to improve the clinical cure rate in humans.

Transcriptional profiling reveals the expression of novel genes in response to various stimuli in the human dermatophyte Trichophyton rubrum

Background: Cutaneous mycoses are common human infections among healthy and immunocompromised hosts, and the anthropophilic fungus Trichophyton rubrum is the most prevalent microorganism isolated from such clinical cases worldwide. The aim of this study was to determine the transcriptional profile of T. rubrum exposed to various stimuli in order to obtain insights into the responses of this pathogen to different environmental challenges. Therefore, we generated an expressed sequence tag (EST) collection by constructing one cDNA library and nine suppression subtractive hybridization libraries. Results: The 1388 unigenes identified in this study were functionally classified based on the Munich Information Center for Protein Sequences (MIPS) categories. The identified proteins were involved in transcriptional regulation, cellular defense and stress, protein degradation, signaling, transport, and secretion, among other functions. Analysis of these unigenes revealed 575 T. rubrum sequences that had not been previously deposited in public databases. Conclusion: In this study, we identified novel T. rubrum genes that will be useful for ORF prediction in genome sequencing and facilitating functional genome analysis. Annotation of these expressed genes revealed metabolic adaptations of T. rubrum to carbon sources, ambient pH shifts, and various antifungal drugs used in medical practice. Furthermore, challenging T. rubrum with cytotoxic drugs and ambient pH shifts extended our understanding of the molecular events possibly involved in the infectious process and resistance to antifungal drugs.

Therapeutic Efficacy of Cintredekin Besudotox (IL13-PE38QQR) in Murine Lung Fibrosis Is Unaffected by Immunity to Pseudomonas aeruginosa Exotoxin A

Background: We have previously explored a therapeutic strategy for specifically targeting the profibrotic activity of IL-13 during experimental pulmonary fibrosis using a fusion protein comprised of human IL-13 and a mutated form of Pseudomonas aeruginosa exotoxin A (IL13-PE) and observed that the intranasal delivery of IL13-PE reduced bleomycin-induced pulmonary fibrosis through its elimination of IL-13-responsive cells in the lung. The aim of the present study was to determine whether the presence of an immune response to P. aeruginosa and/or its exotoxin A (PE) would diminish the anti-fibrotic properties of IL13-PE. Methodology/Principal Findings: Fourteen days after P. aeruginosa infection, C57BL/6 mice were injected with bleomycin via the intratracheal route. Other groups of mice received 4 doses of saline or IL13-PE by either intranasal or intraperitoneal application, and were challenged i.t. with bleomycin 28 days later. At day 21 after bleomycin, all mice received either saline vehicle or IL13-PE by the intranasal route and histopatological analyses of whole lung samples were performed at day 28 after bleomycin. Intrapulmonary P. aeruginosa infection promoted a neutralizing IgG2A and IgA antibody response in BALF and serum. Surprisingly, histological analysis showed that a prior P. aeruginosa infection attenuated the development of bleomycin-induced pulmonary fibrosis, which was modestly further attenuated by the intranasal administration of IL13-PE. Although prior intranasal administration of IL13-PE failed to elicit an antibody response, the systemic administration of IL13-PE induced a strong neutralizing antibody response. However, the prior systemic sensitization of mice with IL13-PE did not inhibit the anti-fibrotic effect of IL13-PE in fibrotic mice. Conclusions: Thus, IL13-PE therapy in pulmonary fibrosis works regardless of the presence of a humoral immune response to Pseudomonas exotoxin A. Interestingly, a prior infection with P. aeruginosa markedly attenuated the pulmonary fibrotic response suggesting that the immune elicitation by this pathogen exerts anti-fibrotic effects.

First Description of a Cluster of Acute/Subacute Paracoccidioidomycosis Cases and Its Association with a Climatic Anomaly

Background: Identifying clusters of acute paracoccidioidomycosis cases could potentially help in identifying the environmental factors that influence the incidence of this mycosis. However, unlike other endemic mycoses, there are no published reports of clusters of paracoccidioidomycosis. Methodology/Principal Findings: A retrospective cluster detection test was applied to verify if an excess of acute form (AF) paracoccidioidomycosis cases in time and/or space occurred in Botucatu, an endemic area in Sao Paulo State. The scan-test SaTScan v7.0.3 was set to find clusters for the maximum temporal period of 1 year. The temporal test indicated a significant cluster in 1985 (P<0.005). This cluster comprised 10 cases, although 2.19 were expected for this year in this area. Age and clinical presentation of these cases were typical of AF paracccidioidomycosis. The space-time test confirmed the temporal cluster in 1985 and showed the localities where the risk was higher in that year. The cluster suggests that some particularities took place in the antecedent years in those localities. Analysis of climate variables showed that soil water storage was atypically high in 1982/83 (similar to 2.11/2.5 SD above mean), and the absolute air humidity in 1984, the year preceding the cluster, was much higher than normal (similar to 1.6 SD above mean), conditions that may have favored, respectively, antecedent fungal growth in the soil and conidia liberation in 1984, the probable year of exposure. These climatic anomalies in this area was due to the 1982/83 El Nino event, the strongest in the last 50 years. Conclusions/Significance: We describe the first cluster of AF paracoccidioidomycosis, which was potentially linked to a climatic anomaly caused by the 1982/83 El Nino Southern Oscillation. This finding is important because it may help to clarify the conditions that favor Paracoccidioides brasiliensis survival and growth in the environment and that enhance human exposure, thus allowing the development of preventive measures.

Evaluating the Taxonomic Identity in Four Species of the Lobose Testate Amoebae Genus Arcella Ehrenberg, 1832

The taxonomic identity in microbial eukaryotes remains an impediment to discussing ecology, biogeography and phylogeny, mainly due to a lack of standards in organism descriptions and few comparative works. The lobose testate amoebae (Arcellinida) present an ideal study system, as progress is severely hindered due to taxonomic confusion. In the present survey, we have examined the morphology, biometry and ecology of 2400 individuals in the genus Arcella Ehrenberg, 1832, collected from the Tiete River in Sao Paulo, Brazil. We then contrasted these new data with 26 previously described species, varieties and forms, looking for consistencies and trying to establish distinct entities. Using a combination of morphology and multivariate statistics we were able to determine 4 distinct taxa (Arcella hemisphaerica, Arcella discoides, Arcella gibbosa and Arcella brasiliensis), each of them encompassing a number of other non-distinct nominal taxa. We describe in detail each of the 4 taxa with notes on ecology and biogeography, and list the indistinguishable names in an effort to make identification and taxonomy in the testate amoebae a more objective and precise exercise by clarifying the taxonomic identity.

Lupeol

The title compound [systematic name: 3 beta-lup-20(29)-en-3-ol], C(30)H(50)O, was isolated from the leaves of Garcinia brasiliensis (common name: bacupari; a member of the Guttiferae family) and has been shown to have many useful medicinal and biological properties. The lupeol molecule consists of four six-membered rings (adopting chair conformations) and one five-membered ring (with an envelope conformation), all fused in trans fashion. Lupeol is isomorphic with the pentacyclic triterpene 3 beta,30-dihydroxylup-20(29)-ene, which differs from lupeol due to the presence of an additional hydroxy group. The crystal packing is stabilized by van der Waals interactions and intermolecular O-H center dot center dot center dot O hydrogen bonds, giving rise to an infinite helical chain along the c axis.

Source populations in coastal crabs: parameters affecting egg production

Benthic marine invertebrates may form metapopulations connected via propagule dispersal. Conservation efforts often target potential source coastlines to indirectly benefit areas depending on allochthonous offspring production. Besides population density, adult size structure, sex ratio, brooding frequency and the proportion of breeding individuals may significantly influence the reproductive output of benthic populations, but these effects have seldom been tested. We used rocky shore crabs to assess the spatial variability of such parameters at relevant scales for conservation purposes and to test their consistency over 2 consecutive years; we then used the data to address whether bottom-up processes or biological interactions might explain the patterns observed. We decomposed egg production rates into their components for the 2 most abundant brachyuran species inhabiting the intertidal rocky habitat. Adult density and brooding frequency varied consistently among shores for both species and largely explained the overall spatial trends of egg production. Temporally consistent patterns also included among-shore differences in the size of ovigerous females of the grapsid Pachygrapsus transversus and between-bay differences in the fecundity of the spider crab Epialtus brasiliensis. Sex ratio was remarkably constant in both. We found no positive or negative correlations between adult density and brooding frequency to support either the existence of a component Allee effect (lack of mate encounters) or an effect of intra-specific competition. Likewise, shore-specific potential growth in P. transversus does not negatively correlate with frequency of ovigerous individuals, as would be expected under a critical balance between these 2 processes. The patterns observed suggest that bottom-up drivers may best explain spatial trends in the reproductive output of these species.

The recent occurrence, establishment and potential impact of Geophagus proximus (Cichlidae : Perciformes) in the Tiete River reservoirs: an Amazonian fish species introduced in the Parana Basin (Brazil)

The introduction of allochthonous fish species happens constantly in large bodies of freshwater, like as the reservoirs of Parana Basin, located in Brazilian southeast, representing a threat for local biodiversity. The fish species Plagioscion squamosissimus and Cichla ocellaris were introduced from the 1970s in several water bodies of this basin and had successfully established themselves in all six reservoirs located in the middle and lower Tiete River (SP, Brazil), particularly. After six decades from the first recorded species introduction, this hydrographic system remains open to the invasion of further fish species, owing to widespread fish-farming activity and by the channels opened between this system and other reservoirs and river basin. This study was an effort to confirm the Geophagus proximus occurrence in the six Tiete River reservoirs, verifying the actual introduction status and analyzing its potential environmental impacts on local species by the analysis of the population structure (abundance, body dimensions and feeding habits). By the results, this species was confirmed in the Ibitinga, Nova Avanhandava and Tres Irmaos reservoirs. The abundance and feeding analysis shows, respectively, it is successfully established in the Tres Irmaos reservoir with the same feeding habitats of local species, such as Geophagus brasiliensis. It was further shown to be very likely that G. proximus would spread throughout the reservoir system of the middle and lower Tiete River, in the manner of P. squamosissimus and C. ocellaris, and the competition pressure for food resources between G. proximus and the local species which represents a potential environmental impact system. These scientific evidences fortifies the knowledge basin for the implantation of a fish management system, to control and reduce the abundance of the invader and to prevent its becoming established in all the Tiete River Basin, avoiding the disastrous consequences for the native species of Parana River Basin.

Oxidative stress biomarkers of exposure in the blood of cichlid species from a metal-contaminated river

The oxidative stress biomarkers of exposure, such as reduced glutathione (GSH), activity of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) and the levels of lipid peroxidation (LPO), were measured in the blood of three cichlid fish (Oreochromis niloticus, Tilapia rendalli, and Geophagus brasiliensis) taken during two seasons from two sites, unpolluted and polluted by industrial effluents, to evaluate the effectiveness of these biomarkers in assessing the impact of water contamination. The LPO levels in the blood were higher in fish from the metal-contaminated site and the chronic exposure led to significant changes in GPx, CAT, and SOD activities in all three cichlid species. The considerable variation of responses in these cichlids to water contamination evidenced differences in sensitivity to the metal contamination and/or in the potential to respond to it highlighting the importance of using a set of related biomarkers to assess the impact of water contamination. (C) 2007 Elsevier Inc. All rights reserved.

Antiproliferative Effect of Benzophenones and their Influence on Cathepsin Activity

The antiproliferative activity of two prenylated benzophenones isolated from Rheedia brasiliensis. the tri-prenylated garciniaphenone and the tetraprenylated benzophenone 7-epiclusianone, was investigated against human cancer cell lines. The antiproliferative activity on melanoma (UACC-62), breast (MCF-7), drug-resistant breast (NCI-ADR), lung/non-small cells (NCI460), ovarian (OVCAR 03), prostate (PC03), kidney (786-0), lung (NCI-460) and tongue (CRL-1624 and CRL-1623) cancer cells was determined using spectrophotometric quantification of the cellular protein content. The effect of these benzophenones on the activity of cathepsins B and G was also investigated. Garciniaphenone displayed cytostatic activity in all cell lines, whereas 7-epiclusianone showed a dose-dependent cytotoxic effect. The IC(50) values for cell proliferation revealed that 7-epiclusianone is more active than garciniaphenone against most of the cell lines. Furthermore, the antiproliferative effects demonstrated by garciniaphenone and 7-epiclusianone were related to their cathepsin inhibiting properties. In conclusion, 7-epiclusianone is a promising naturally occurring agent which displays multiple inhibitory effects which may be working in concert to inhibit cancer cell proliferation in vitro. The putative pathway by which 7-epiclusianone affects cancer cell development may involve cathepsin inhibition. Copyright (C) 2009 John Wiley & Sons, Ltd.

Effects of 7-Epiclusianone on Streptococcus mutans and Caries Development in Rats

The aim of this study was to evaluate the effects of 7-epiclusianone (7-epi) on specific virulence attributes of Streptococcus mutans in vitro and on development of dental caries in vivo. 7-Epi was obtained and purified from fruits of Rheedia brasiliensis. We investigated its influence on surface-adsorbed glucosyltransferase (Gtf) B activity, acid production, and viability of S. mutans in biofilms, as well as on caries development using a rodent model. 7-Epi (100 mu g/mL) significantly reduced the activity of surface-adsorbed GtfB (up to 48.0 +/- 1.8 of inhibition at 100 mu g/mL) and glyco-lytic pH-drop by S. mutans in biofilms (125 and 250 mu g/mL) (vs. vehicle control, p < 0.05). In contrast, the test compound did not significantly affect the bacterial viability when compared to vehicle control (15% ethanol, p > 0.05). Wistar rats treated topically with 7-epi (twice daily, 60-s exposure) showed significantly smaller number of and less severe smooth-and sulcal-surface carious lesions (p < 0.05), without reducing the S. mutans viable population from the animals` dental biofilms. In conclusion, the natural compound 7-epiclusianone may be a potentially novel pharmacological agent to prevent and control dental caries disease.

New approaches to underground systems in Brazilian Smilax species (Smilacaceae)

MARTINS, A. R. (Institute of Biology, State University of Campinas - UNICAMP, 13083-970, Campinas, SP, Brazil), N. PUT, (Division of Biology and Education, University of Vechta, 49377 Vechta, Germany), A. N. SOARES, A.B BOMB, and B. APPEZZATO DA GLORIA (Biological Science Department, Escola Superior de Agricultura `Luiz de Queiroz`, University of Sao Paulo, 13418-900, Piracicaba, SP, Brazil). J. Torrey Bot. Soc. 137: 220-235. 2010.-New approaches to underground systems in Brazilian Smilax species (Smilacaceae). Scientific studies show that the watery extract of the thickened underground stem and its adventitious roots of the genus Smilax can act as a therapeutic agent in immunoinflammatory disorders, such as rheumatic arthritis. Brazilians have used this genus of plants in folk medicine, however it is very hard to identify these species, since the morphology of the underground systems is very similar in this group. For better identification of those systems, we studied six species of Smilax L. (S. brasiliensis, S. campestris, S. cissoides, S. goyazana, S. oblongifolia and S. rufescens), collected in different regions of Brazil with different physiognomies and soil characteristics. The main purpose is to describe the morpho-anatomy of the underground systems and to analyze if their structure depends on environmental conditions. The underground stem (rhizophore) is of brown color and it is knotty, massive, slender (S. rufescens) or tuberous (S. brasiliensis, S. campestris, S. cissoides, S. goyazana and S. oblongifolia). The tuberization is a result of primary thickened meristem (PTM) activity. The color and thickness of the adventitious roots change during development because the epidermis and outer cortex are disposed of, so the inner cortex becomes the new covering tissue with lignified and dark color cells. There are differences in starch grain shapes in mature roots. The chemical attributes of the soil are very similar in all studied environments and, even when soil characteristics varied, all the species` underground system was distributed close to the soil surface (10 to 15 cm deep). The species exhibited clonal growth hence their underground system functions as storage structures and the axillary buds can sprout into new stems. Only Smilax rufescens, collected in sandy soil of Restinga, has vegetative dispersal due to the runners.

Rhodacaridae mites (Acari: Mesostigmata: Rhodacaroidea) from the state of Sao Paulo, Brazil, with descriptions of a new genus and three new species

- This paper reports the occurrence of five species of Rhodacaridae mites collected in the State of Sao Paulo, Brazil. One of these corresponds to a new genus and a new species, Binodacarus brasiliensis n. gen. n. sp.; two correspond to new species of kno