Cryptosporidium sp. in children suffering from acute diarrhea at Uberlândia City, State of Minas Gerais, Brazil

This study's objective was to search for Cryptosporidium sp. in diarrheic feces from children aged zero to 12 years and cared for at medical units within Universidade Federal de Uberlândia or at a private practice in Uberlândia, State of Minas Gerais, Brazil, from September 1992 to August 1993. Three fecal samples preserved in 10% formalin, were collected from 94 children. Oocyst concentration was performed through Ritchie's (modified) method and staining of fecal smears for each sample (total of 1128 slides) was done by the "Safranin/Methylene Blue" and the "Kinyoun (modified)" techniques. The Hoffmann, Pons & Janer method was also employed to look for other enteroparasites. From 94 children, 4.26% excreted fecal Cryptosporidium oocysts. The infection seemed to vary according to age: 5.08% of patients aged zero to two years old; 33.33% of those aging eight to ten years (P>0.05). Cryptosporidium appeared in November, December and March, during the rainy season. 20.21% of the children harbored at least one enteroparasite different from Cryptosporidium, mainly Giardia intestinalis (12.77%). From Cryptosporidium infected patients, two had only this kind, another harbored Giardia intestinalis; the last one hosted Strongyloides stercoralis

Intestinal parasites in school food handlers in the city of Uberlândia, Minas Gerais, Brazil

In order to verify the presence of intestinal parasites in food handlers, stool samples were collected from 104 cooks and their helpers that were working in food preparation in 20 public elementary schools, in various areas of the city of Uberlândia, Minas Gerais, Brazil. The samples were collected during the months of November and December, 1988, in plastic flasks containing a 10% formaldehyde solution and processed by the Hoffmann, Pons & Janer method. The sediment was examined using triplicate slides. All individuals were females aged between 24 to 69 years. Intestinal parasites were found in 85.0% of the studied schools and 47.1% of the studied food handlers (cooks and helpers) were found to be positive. Among the 49 infected food handlers, 32 (65.3%) carried a single parasite and 17 (34.7%) carried two parasites. The following intestinal parasites were found: Giardia lamblia (21.1%), Entamoeba coli (21.1%), hookworms (9.6%), Ascaris lumbricoides (5.8%), Entamoeba histolytica (2.9%), Hymenolepis nana (1.9%), Strongyloides stercoralis (1.0%). These data emphasize the need for a rigid semi-annual control in all school food handlers, including diagnosis, specific treatment and orientation about the mechanisms of transmission of the intestinal parasites.Para verificar a presença de parasitas intestinais em manipuladores de merenda escolar foram coletadas amostras fecais de 104 cantineiras e serventes que auxiliavam no preparo da alimentação de 20 Estabelecimentos da rede pública de ensino de 1º grau, localizados em diferentes bairros, da cidade de Uberlândia, Minas Gerais. As amostras foram coletadas nos meses de novembro e dezembro de 1988 em frascos plásticos contendo formol a 10% e processadas pelo método de Hoffmann, Pons & Janer sendo o sedimento examinado em triplicata. Todos os indivíduos pertenciam ao sexo feminino com idade entre 24 e 69 anos. Foram detectados manipuladores de merenda escolar parasitados em 85% das Escolas estudadas. Das 104 amostras de fezes, 49 (47,1%) estavam positivas, sendo que 32 (65,3%) pertenciam a indivíduos monoparasitados e 17 (34,7%) a indivíduos biparasitados. Os parasitas em ordem decrescente de ocorrência foram: Giardia lamblia (21,1%), Entamoeba coli (21,1%), ancilostomídeos (9,6%), Ascaris lumbricoides (5,8%), Entamoeba histolytica (2,9%), Hymenolepis nana (1,9%) e Strongyloides stercoralis (1,0%). Conclui-se pela necessidade de rigoroso controle semestral, tratamento específico e orientação sobre os mecanismos de transmissão das enteroparasitoses a todos os manipuladores de merenda escolar dos Estabelecimentos de ensino

Eutirucallin, a RIP-2 Type Lectin from the Latex of <i>Euphorbia tirucalli</i> L. Presents Proinflammatory Properties

<div><p>Lectins are carbohydrate-binding proteins that recognize and modulate physiological activities and have been used as a toll for detection and identification of biomolecules, and therapy of diseases. In this study we have isolated a lectin present in the latex of Euphorbia tirucalli, and named it Eutirucallin. The latex protein extract was subjected to ion exchange chromatography and showed two peaks with haemagglutinating activity. Polypeptides of 32 kDa protein extract strongly interacted with immobilized galactose (α-lactose > D-N-acetylgalactosamine). The Eutirucallin was obtained with a yield of 5.6% using the α-lactose column. The lectin domain has 32 kDa subunits and at least two of which are joined by disulfide bridges. The agglutinating capacity for human erythrocytes A⁺, B⁺ and O⁺ is inhibited by D-galactose. The haemagglutinating activity of Eutirucallin was independent of Ca²⁺ and maintained until the temperature of 55°C. Eutirucallin presented biological activities such as neutrophils recruitment and cytokine prodution by macrophages. The analysis of the trypsin-digested Eutirucallin by ms/ms in ESI-Q-TOFF resulted in nine peptides similar to type 2 ribosome-inactivating protein (type-2 RIP). It's partial sequence showed a similarity of 67.4 – 83.1% for the lectin domain of type-2 RIP [Ricin and Abrin (83.1%), Viscumin, Ebulin, Pulchellin, Cinnamomin, Volkensin and type-2 RIP Iris hollandica]. Our data suggest that Eutirucallin is a new member of type 2 ribosome-inactivating protein and presents biotechnological potential.</p></div

Lectin purification.

a<p>A unit is defined as the minimum protein concentration required, able to promote the complete agglutination of erythrocytes (title⁻¹). ^bThe specific haemagglutinating activity is defined by the ratio between the UH and mg of protein used. Human erythrocytes A⁺ were used in the assay.</p

Alignment of partial sequence of Eutirucallin with type 2 ribosome-inactivating protein.

<p>All identity and similarity analyses were performed with the tool clustalw2. The sequences used were: Ricin (GenBank: XP_002534649.1), Abrin a (PDB: 2AAI_B), Viscumin (PDB: 2RG9_B), Ebulin (PDB: 1HWM_B), Pulchellin (GenBank: AAM19073.1), Cinnamomin (GenBank: AAK82460.1), Volkensin (GenBank: CAD61022.1) and type 2 RIP of <i>Iris hollandica</i> (GenBank: AAL55093.1). The letters “single letter code” correspond to amino acids residues. (*) Represent identical residues; ( ) residues identical for at least one of the sequences; (:) conserved residues; and (.) semi-conserved replacements. The arrow (↓) corresponds to the initial residue of B-chain (lectin domain). The residues involved in the glycidic recognition are marked with closed circles (•); the probable glycosylation points are represented by open squares (□).</p

Comparative analysis of similarity/identity of the Eutirucallin sequence with eight B-chains of type 2 ribosome- inactivating protein.

a<p>The identity amounts are presented below the axis (----) and represents the percentage of identical amino acids residues. Similarity amounts are presented above the diagonal axis. ^bThe amounts were obtained by considering only the paired regions of the polypeptide sequence of the Eutirucallin. All analyses were made with the tool Clustalw2.</p

Construction of the polypeptide sequence of Eutirucallin.

<p>A. Cluster assembled from of peptides obtained by ms/ms. B. The order of peptides in the cluster was established by alignment with ricin. The letters “single letter code” correspond to the amino acids residues. (*) Represent identical residues; (:) conserved residues; and (.) semi-conserved replacements. The arrow (↓) corresponds to the initial residues of the A- (N-glycosidase) and B- (lectin domain) chains, respectively. The signal peptide is indicated in the sequence initial segment. The residues (cys-cys) correspond to the intramolecular and intermolecular disulfide bindings, indicated by the full and dashed lines, respectively.</p

Influence of the heat treatment and EDTA on the haemagglutinating activity of the Eutirucallin.

<p><b>A.</b> Eutirucallin (1.8 µg) was incubated for 20 min at 25, 35, 45, 55, 65, 75, 85 and 95°C. After 60 min of sedimentation at room temperature the haemagglutinating activity was evaluated. <b>B.</b> The Eutirucallin (1.8 µg) was incubated with EDTA in the concentrations of 10, 50 and 100 mM, and kept for 18 h at 4°C. Following to that the haemagglutinating activity was evaluated. Human erythrocytes A+ at 2% were used in both assays. The positive haemagglutination was indicated by the formation of a uniform coating of red blood cells in the surface of the U plates, and its absence was determined by the sedimentation of the cells in the bottom of the plates.</p