Ary Corrêa

Infection with Strongyloides venezuelensis Induces Transient Airway Eosinophilic Inflammation, an Increase in Immunoglobulin E, and Hyperresponsiveness in Rats

ABSTRACT Infection by nematode parasites with a pulmonary migration in their life cycle and allergic asthma are two highly prevalent diseases in humans; therefore, one may expect both may occur concomitantly. There is a predominant and essential role of Th2 lymphocytes in the mechanisms underlying the control of parasite elimination as well as in the pathology observed in the asthmatic lung. The consequences of such situations have been explored, with controversial results, justifying the development of experimental models in which the relationship between allergic airway inflammation and helminth infection might be evaluated. The present work describes the inflammatory, humoral, and functional changes that occur in the lung of rats after single (subcutaneous inoculation of 1,500 L3 larvae) or multiple (five weekly subcutaneous inoculations of 1,500 L3 larvae) Strongyloides venezuelensis infections. The results show that the migration of S. venezuelensis larvae through the lungs of infected rats induces a local eosinophilic inflammation process which is mostly focal and parenchymal for rats infected a single time and which is peribronchial after multiple infections. The inflammatory process is accompanied by mucus hypersecretion, thickening of bronchial epithelial and muscle layers, and local increase in immunoglobulin E concentrations that peak after 5 to 7 days and are resolved after 12 days of single or multiple infections. The peak of lung immunopathologic changes observed in infected rats coincides with lung airway hyperresponsiveness (AHR), a key functional alteration in asthma. We propose that this experimental model is ideal to carry out further studies on immunoprotection against nematode infection versus immunopathology of allergic airway inflammation.

Role of the bradykinin B2 receptor for the local and systemic inflammatory response that follows severe reperfusion injury

Bradykinin (BK) appears to play an important role in the development and maintenance of inflammation. Here, we assessed the role of the BK B2 receptor for the injuries that occur after ischemia and reperfusion (I/R) of the territory irrigated by the superior mesenteric artery. Tissue (lung and duodenum) kallikrein activity increased after ischemia with greater enhancement after reperfusion. A selective inhibitor of tissue kallikrein, Phenylacetyl‐Phe‐Ser‐Arg‐N‐(2,3‐dinitrophenyl)‐ethylenediamine (TKI, 0.001–10 mg ml−1), inhibited kallikrein activity in a concentration‐dependent manner in vitro. In vivo, pretreatment with TKI (30 mg kg−1) prevented the extravasation of plasma and the recruitment of neutrophils. Similarly, the bradykinin B2 receptor antagonists, HOE 140 (0.01–1.0 mg kg−1) or FR173657 (10.0 mg kg−1), inhibited reperfusion‐induced increases in vascular permeability and the recruitment of neutrophils in the intestine and lungs. In a model of more severe I/R injury, HOE 140 (1.0 mg kg−1) inhibited the increase in vascular permeability, neutrophil recruitment, haemorrhage and tissue pathology. Furthermore, HOE 140 significantly inhibited the elevations of TNF‐α in tissue and serum and partially prevented lethality. This was associated with an increase in the concentrations of IL‐10 in tissue and serum. Thus, our results demonstrate that, following intestinal I/R injury, there is an increase in tissue kallikrein activity and activation of BK B2 receptors. B2 receptor activation is essential for the development of inflammatory tissue injury and lethality. These results contrast with those of others showing that BK mostly exerts a protective role during I/R injury.

Diversity and frequency of yeasts from the dorsum of the tongue and necrotic root canals associated with primary apical periodontitis

AIMS To determine the frequency and diversity of yeasts from the dorsum of the tongue and necrotic root canals with teeth associated with primary apical periodontitis. METHODOLOGY Detailed medical and dental histories of 168 patients were recorded. The samples were collected from the dorsum of tongue and from 168 teeth with root canals contained necrotic pulps. Yeasts were isolated on Sabouraud agar with 100 mg L(-1) chloramphenicol, purified and characterized by standard methods. Identification was confirmed by EI1 PCR fingerprint technique. Yeast isolates of uncertain identity or with a different genetic fingerprint profile from the reference strains were identified by sequencing the D1/D2 variable domains of the large subunit rDNA. RESULTS Yeasts were isolated from 22.6% of teeth sampled and from 45.8% of tongue samples. Candida albicans was the most frequently isolated species at both investigated sites but other species were also found. Saccharomyces cerevisiae and Kluyveromyces lactis were recovered from the tongue. CONCLUSIONS Although the detection of yeasts in the root canal does not imply an involvement in the disease process, the study suggests a frequency of Candida spp. in primary endodontic infections that deserves further clarification.

Differential lectin labelling of circulating hemocytes from Biomphalaria glabrata and Biomphalaria tenagophila resistant or susceptible to Schistosoma mansoni infection

Lectins/carbohydrate binding can be involved in the Schistosoma mansoni recognition and activation of the Biomphalaria hemocytes. Therefore, expression of lectin ligands on Biomphalaria hemocytes would be associated with snail resistance against S. mansoni infection. To test this hypothesis, circulating hemocytes were isolated from B. glabrata BH (snail strain highy susceptible to S. mansoni), B. tenagophila Cabo Frio (moderate susceptibility), and B. tenagophila Taim (completely resistant strains), labelled with FITC conjugated lectins (ConA, PNA, SBA, and WGA) and analyzed under fluorescence microscopy. The results demonstrated that although lectin-labelled hemocytes were detected in hemolymph of all snail species tested, circulating hemocytes from both strains of B. tenagophila showed a larger number of lectin-labelled cells than B. glabrata. Moreover, most of circulating hemocytes of B. tenagophila were intensively labelled by lectins PNA-FITC and WGA-FITC, while in B. glabrata small hemocytes were labeled mainly by ConA. Upon S. mansoni infection, lectin-labelled hemocytes almost disappeared from the hemolymph of Taim and accumulated in B. glabrata BH. The role of lectins/carbohydrate binding in resistance of B. tengophila infection to S. mansoni is still not fully understood, but the data suggest that there may be a correlation to its presence with susceptibility or resistance to the parasite.

Participation of cell‐free haemolymph of Biomphalaria tenagophila in the defence mechanism against Schistosoma mansoni sporocysts

Biomphalaria tenagophila of Taim strain is able to completely destroy Schistosoma mansoni sporocyst few hours after parasite penetration, although the mechanism is still not well known. In this experimental work we show that passive transference of cell‐free haemolymph, especially from B. tenagophila Taim, resulted in higher resistance of B. tenagophila Cabo Frio to S. mansoni infection. This effect was demonstrated in vivo, by the reduction in the infection rate, and the significantly lower production of sporocysts and cercariae of the parasite in snails treated with Taim cell‐free haemolymph compared to CBSS‐inoculated snails. The protective effect of Taim cell‐free haemolymph was also observed during the in vitro interaction between haemocytes and sporocysts. In this system, addition of B. tenagophila cell‐free haemolymph, especially from Taim strain, was responsible for significant increase in sporocyst mortality compared to B. glabrata cell‐free haemolymph or culture medium. Moreover, the combination of Taim cell‐free haemolymph and Cabo Frio haemocytes increased significantly the mortality of sporocysts. The results show that Taim cell‐free haemolymph would act direct and indirectly on destruction of S. mansoni sporocysts. The results also suggest that cell‐free haemolymph indirectly increases parasite recognition by the circulating granulocytes and it is species specific.

Experimental Infection with Schistosoma mansoni in CCR5-Deficient Mice Is Associated with Increased Disease Severity, as CCR5 Plays a Role in Controlling Granulomatous Inflammation

ABSTRACT The plasma level of the chemokine CCL3 is elevated in patients with chronic severe schistosomiasis mansoni. We have previously shown that CCL3−/− mice with experimental infection showed diminished pathology and worm burden compared to those of wild-type (WT) mice. To elucidate further the role of CC chemokines during schistosomiasis mansoni infection, we evaluated the course of infection in C57BL/6J mice deficient in CCR5, one of the receptors for CCL3. The CCR5 deficiency proved to be remarkably deleterious to the host, since mortality rates reached 70% at 14 weeks postinfection in CCR5−/− mice and 19% in WT mice. The increased lethality was not associated with an increased parasite burden, since similar numbers of eggs and adult worms were found in mice from both groups. Liver granulomas of chronically infected CCR5−/− mice were larger and showed greater numbers of cells and collagen deposition than liver granulomas from WT mice. This was associated with higher levels of production of intereleukin-5 (IL-5), IL-13, CCL3, and CCL5 in infected CCR5−/− mice than in infected WT mice. Moreover, at 8 weeks after infection, just before changes in pathology and mortality, the numbers of FoxP3-positive cells were lower in liver granulomas of CCR5−/− mice than in WT mice. In conclusion, the CCR5 deletion is deleterious to mice infected with Schistosoma mansoni, and this is associated with enhanced fibrosis and granulomatous inflammation.

Integrated control of Penicillium digitatum by the predacious yeast Saccharomycopsis crataegensis and sodium bicarbonate on oranges

Our investigation of integrated biological control (IBC) started with an assay testing activity of the predacious yeast Saccharomycopsis crataegensis UFMG-DC19.2 against Penicillium digitatum LCP 4354, a very aggressive fungus that causes postharvest decay in oranges. Under unfavourable environmental conditions, the yeast showed a high potential for control (39.9% disease severity reduction) of this fungus. This result was decisive for the next step, in which S. crataegensis was tested in association with sodium bicarbonate salt, a generally regarded as safe (GRAS) substance. The yeast was able to survive at different concentrations of the salt (1%, 2% and 5%), and continued to grow for a week at the wound site, remaining viable at high population for 14 days on the fruit surface. The yeast alone reduced the severity of decay by 41.7% and sodium bicarbonate alone reduced severity of decay by 19.8%, whereas the application of both led to a delay in the development of symptoms from 2 to 10 days. Ingredients of the formulations were not aggressive to fruits since no lesions were produced in control experiments.

Microinjected antisense Inf24 oligonucleotides inhibit appressorium development in Uromyces

Germlings of Uromyces appendiculatus respond to topographical stimuli and develop appressoria. During this period several genes are expressed, one of which is Inf 24. Germlings were microinjected with a sense and antisense fragment of the ORF of Inf 24. Sense ( Inf 24-S), 5′-ATG AGT GCT TCA TCG CAG CAG CAG-3′ fragment did not affect germling response to the topographical stimuli and the developed appressoria. Microinjection of antisense ( Inf 24-RC), 5′-CTG CTG CTG CGA TGA AGC ACT CAT-3′ blocked gene transcription and appressoria were rarely formed. Injection of Inf 24-RC into already formed appressoria did not inhibit continued development of subsequent infection structures, e.g. penetration peg, sub-stomatal vesicle.

Participation of N-acetyl-D-glucosamine carbohydrate moieties in the recognition of Schistosoma mansoni sporocysts by haemocytes of Biomphalaria tenagophila

Lectin-carbohydrate binding may be involved in the recognition of Schistosoma mansoni sporocysts by haemocytes of Biomphalaria; therefore, we tested if this interaction is associated with snail resistance against Schistosoma infection. In vitro data showed that most of the S. mansoni sporocysts cultured with haemocytes from Biomphalaria glabrata BH, a highly susceptible snail strain, had a low number of cells that adhered to their tegument and a low mortality rate. Moreover, the addition of N-acetyl-D-glucosamine (GlcNAc) did not alter this pattern of adherence and mortality. Using haemocytes and haemolymph of Biomphalaria tenagophila Cabo Frio, we observed a high percentage of sporocysts with adherent cells, but complete encapsulation was not detected. Low concentrations of GlcNAc increased haemocyte binding to the sporocysts and mortality, which returned to basal levels with high concentrations of the carbohydrate. In contrast, haemocytes plus haemolymph from B. tenagophila Taim encapsulated cellular adhesion index of level 3 and destroyed over 30% of the S. mansoni sporocysts in culture. Interestingly, the addition of GlcNAc, but not mannose, to the culture medium resulted in the significant inhibition of cellular adhesion to the parasite tegument and the reduction of parasite mortality, suggesting that GlcNAc carbohydrate moieties are important to the recognition of S. mansoni by B. tenagophila Taim.

YEAST COMMUNITIES IN TWO ATLANTIC RAIN FOREST FRAGMENTS IN SOUTHEAST BRAZIL

We studied the yeast communities associated with fruits, mushrooms, tree exudates, and flies of the genus Drosophila, in two Atlantic Rain Forest fragments in state of Minas Gerais, Brazil. A total of 456 samples were collected from Rio Doce State Park and 142 from Ecological Station of Universidade Federal de Minas Gerais. From these samples, 608 yeast isolates were obtained, belonging to 71 different species. Among the yeasts isolated from Rio Doce State Park, 17 isolates were recovered from fruits, 12 from mushrooms, 13 from tree exudates, and 299 from Drosophila spp. In the Ecological Station of Universidade Federal de Minas Gerais, 24 isolates were recovered from fruits and 243 from Drosophila spp. Distinct communities of yeast were observed in Drosophila flies, fruits, mushrooms and tree exudates. The highest number of yeast species was recovered from Drosophila flies suggesting that flies are the natural vectors of these microorganisms.