Relationship of intestinal histology with the resistance to Trichostrongylus colubriformis infection in three breeds of sheep

Avaliaram-se a associação entre o número de células inflamatórias no intestino delgado e a resistência à infecção por Trichostrongylus colubriformis em ovinos de três raças (Santa Inês, Suffolk e Ile de France), naturalmente infectados. Mastócitos, eosinófilos e leucócitos globulares foram quantificados na mucosa intestinal. A concentração de histamina foi estimada em amostras teciduais do intestino, bem como foi determinado o comprimento de machos e fêmeas de T. colubriformis. A resposta celular foi similar na mucosa intestinal das três raças ovinas (P>0,05). Houve grande variação entre os ovinos em relação aos resultados parasitológicos e celulares, mesmo nos animais de mesma raça. em geral, os animais que apresentaram número menor de células inflamatórias tiveram cargas parasitárias maiores, contagens de ovos por grama de fezes mais altas e exemplares de T. colubriformis maiores. Os resultados indicaram que mastócitos, eosinófilos e leucócitos globulares prejudicaram o estabelecimento, o desenvolvimento e a sobrevivência dos parasitas.

Comparison of three concentration methods for the recovery of canine intestinal parasites from stool samples

The aim of present study was to compare the efficiency of a commercial assay and two conventional methods for fecal concentration in detecting canine gastrointestinal parasites. Fecal samples from 254 dogs were processed by centrifugation-sedimentation (CS), centrifugation-flotation (CF) and a commercial assay for fecal concentration (TF-test (R)). The following parasites were detected: Ancylostoma (37.8%), Giardia (16.9%), Toxocara canis (8.7%), Trichuris vulpis (7.1%), Isospora (3.5%), and Sarcocystis (2.7%). The calculated analytical sensitivity indicated that CF was more accurate (P < 0.01) in detecting Ancylostoma, T. canis, T. vulpis and Giardia infections. However, CF showed significantly higher sensitivity only for Ancylostoma, compared to the other two methods. The kappa index value of diagnostic agreement between TF-test and CF was high for T. canis (83%) and moderate for Giardia (72%) and Ancylostoma (63%). The advantages and limitations of each method were assessed for individual diagnosis and epidemiological investigation. (C) 2010 Elsevier B.V. All rights reserved.

Diagnosis of Giardia infections by PCR-based methods in children of an endemic area

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Identification and characterization of excretory/secretory products of Giardia duodenalis trophozoites

One of the major questions concerning Giardia is the understanding of pathophysiological processes associated with small intestine abnormalities. There are evidences that Giardia trophozoites contain and/or release proteolytic enzymes that may be implicated in the host intestinal epithelium. The present investigation was undertaken to examine the protease activity in excretory/secretory (E/S) products of Giardia duodenalis trophozoites of an axenic Brazilian strain (BTU-11) and the reference strain Portland 1 (P1). E/S products from trophozoites of each strain in conditioned medium were tested with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) for the protein profiles, and the protease activity was analyzed using substrate-impregnated SDS-PAGE (gelatin and collagen) and hemoglobin assay. The proteases characterization was based on inhibition assays including synthetic inhibitors. Electrophoresis analysis of E/S products revealed a banding pattern composed by few bands (4 to 6 bands) in the migration region of 123 to 28 kDa. Proteolytic products were detected in the conditioned medium by trophozoites of both assayed strains. In the gels containing copolymerized gelatin and collagen, E/S products promoted substrate degradation and the most evident proteolysis zones were distributed in the migration regions of 77 to 18 kDa and 145 to 18 kDa, respectively, in the patterns of gelatinolytic and collagenolytic activities. Degradation of hemoglobin was also observed, and the pattern of hydrolysis was similar in both E/S products assayed. Inhibitor assays showed that the main proteolytic activity in both E/S products is due to cysteine proteases, although the presence of serine proteases was also indicated. Degradation of substrates including collagen and hemoglobin could lead us to speculate different functions of Giardia excreted/secreted proteases in vivo, but to confirm this possibility and to elucidate its implication on host-parasite interactions, further experiments applying protocols for the purification of proteases are necessary. Even so, our observations are relevant and hold the perspective for the understanding about protease activity in Giardia trophozoites of axenic strain isolated in an endemic area.

The use of biodiversity as source of new chemical entities against defined molecular targets for treatment of malaria, tuberculosis, and T-cell mediated diseases: a review

The modern approach to the development of new chemical entities against complex diseases, especially the neglected endemic diseases such as tuberculosis and malaria, is based on the use of defined molecular targets. Among the advantages, this approach allows (i) the search and identification of lead compounds with defined molecular mechanisms against a defined target (e.g. enzymes from defined pathways), (ii) the analysis of a great number of compounds with a favorable cost/benefit ratio, (iii) the development even in the initial stages of compounds with selective toxicity (the fundamental principle of chemotherapy), (iv) the evaluation of plant extracts as well as of pure substances. The current use of such technology, unfortunately, is concentrated in developed countries, especially in the big pharma. This fact contributes in a significant way to hamper the development of innovative new compounds to treat neglected diseases. The large biodiversity within the territory of Brazil puts the country in a strategic position to develop the rational and sustained exploration of new metabolites of therapeutic value. The extension of the country covers a wide range of climates, soil types, and altitudes, providing a unique set of selective pressures for the adaptation of plant life in these scenarios. Chemical diversity is also driven by these forces, in an attempt to best fit the plant communities to the particular abiotic stresses, fauna, and microbes that co-exist with them. Certain areas of vegetation (Amazonian Forest, Atlantic Forest, Araucaria Forest, Cerrado-Brazilian Savanna, and Caatinga) are rich in species and types of environments to be used to search for natural compounds active against tuberculosis, malaria, and chronic-degenerative diseases. The present review describes some strategies to search for natural compounds, whose choice can be based on ethnobotanical and chemotaxonomical studies, and screen for their ability to bind to immobilized drug targets and to inhibit their activities. Molecular cloning, gene knockout, protein expression and purification, N-terminal sequencing, and mass spectrometry are the methods of choice to provide homogeneous drug targets for immobilization by optimized chemical reactions. Plant extract preparations, fractionation of promising plant extracts, propagation protocols and definition of in planta studies to maximize product yield of plant species producing active compounds have to be performed to provide a continuing supply of bioactive materials. Chemical characterization of natural compounds, determination of mode of action by kinetics and other spectroscopic methods (MS, X-ray, NMR), as well as in vitro and in vivo biological assays, chemical derivatization, and structure-activity relationships have to be carried out to provide a thorough knowledge on which to base the search for natural compounds or their derivatives with biological activity.

Activity of selenium on cell proliferation, cytotoxicity, and apoptosis and on the expression of CASP9, BCL-XL and APC in intestinal adenocarcinoma cells

Intestinal cancers are correlated with diet. Thus, determining and understanding nutrient-genome interactions is important. The present work assessed the action of the oligoelement selenium on cell proliferation, cytotoxicity, and in situ apoptosis induction and on the expression CASP9, BCL-XL and APC genes in intestinal adenocarcinoma cells (HT29). HT29 cells were cultured and treated with selenium at concentrations of 5, 50 and 500 ng/mL with or without the damage-inducing agent doxorubicin. These cells were then evaluated for cytotoxicity (MTT), cell proliferation and in situ apoptosis induction. To evaluate gene expression, only the cells treated with 500 ng/mL of selenium were used. RNA was extracted from these cells, and the expressions of CASP9, BCL-XL and APC were analyzed by the RT-PCR method. The GAPDH gene was used as a reference gene. The MTT assay showed that selenium was not cytotoxic at any of the concentrations tested. The cell proliferation assay showed that selenium did not interfere with cell proliferation at the three concentrations tested. In contrast, when the three concentrations were combined with doxorubicin, a significant decrease in the proliferation rate was observed. The apoptosis rate was significantly increased in the selenium (500 ng/mL) and doxorubicin group. CASP9 expression was increased and BCL-XL expression decreased in the selenium (500 ng/mL) and doxorubicin group. APC was significantly increased in the selenium group alone. These results show that selenium increases apoptosis, especially when it is associated with a damage-inducing agent. Also, selenium has an important role in the expression of the APC gene, which is related to cell cycle regulation. (C) 2011 Elsevier B.V. All rights reserved.

Early physical activity promotes lower prevalence of chronic diseases in adulthood

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Emerging infectious diseases in cetaceans worldwide and the possible role of environmental stressors

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Skin diseases in Guiana dolphins (Sotalia guianensis) from the Paranagua estuary, Brazil: A possible indicator of a compromised marine environment

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Obstrução intestinal e enterotomia em tigre d’água (Trachemys dorbignyi)

Intestinal obstruction occurs in chelonian mainly due parasitism and foreign bodies, as stones and sand. An intestinal compaction was described in a five year-old male d'orbigny's slider which was taken to the Veterinary Medical Teaching Hospital of the UNICASTELO, at Fernandopolis, SP, Brazil, presenting anorexia for a week, severe dehydration and stupor. Definitive diagnostic was performed by radiographic exam and great amount of intestinal radiopac substance was detected. The animal went through emergency celiotomy for removing the intestinal foreign bodies. Inhalatory anesthesia with isofluorane was used for anesthesia induction and manutention. After local antisepsy, a 4cm(2) oblique opening was conducted on plastron by using a vibratory saw. Peritoneum was cut, intestines exteriorized and enterotomy performed. During the surgery, the animal was radiographed to confirm the complete taken out of the foreign bodies. Mononylon 4-0 strand was applied for intestinal suture in two planes. The plastron piece that was taken out was replaced and set with epoxy resine and gaze on the surgical window, making it waterproof. In the postoperative time, animal was medicated with analgesics for two days and pentabiotics for five days. Oral creamy diet was used with oral tube from the second to tenth day, what provided a great clinicosurgical recovering.

Avaliação química e da atividade antidiarréica das folhas de Byrsonima cinera DC. (Malpighiaceae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Relative antioxidant activity of Brazilian medicinal plants for gastrointestinal diseases

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Conseqüências nutricionais das alterações metabólicas dos macronutrientes na doença hepática crônica

A doença hepática crônica cursa, freqüentemente, com anormalidades metabólicas de macronutrientes que propiciam o desenvolvimento ou agravamento da desnutrição protéico-energética. O papel central do fígado no metabolismo dos substratos energéticos e de proteínas e aminoácidos é revisto, de modo relacionado à desnutrição protéico-energética, em pacientes com hepatopatia crônica. Aceita-se que a redução da ingestão dietética seja um dos principais componentes etiológicos da desnutrição, particularmente em pacientes alcoolistas. Acresce-se a iatrogenia pela indicação de dietas restritas e jejum prolongado aos pacientes hospitalizados. Como fatores agravantes, há má absorção intestinal de gorduras e o hipermetabolismo associado ao alcoolismo agudo. Hipoglicemia, resistência insulínica, esteatose e hipertrigliceridemia constituem achados comuns, assim como níveis elevados de alguns aminoácidos com conseqüências neurológicas. O entendimento desses mecanismos fisiopatológicos permite a intervenção nutricional apropriada reduzindo a morbidade e mortalidade desses pacientes.

Implicações do alcoolismo e da doença hepática crônica sobre o metabolismo de micronutrientes

A doença hepática, alcoolismo e desnutrição são condições comumente associadas que interferem no metabolismo de micronutrientes. Como resultado da doença hepática pode ocorrer menor estocagem e conversão de vitaminas nas suas formas ativas, e má digestão e/ou má absorção. Há ainda o agravante do álcool diminuindo a ingestão e absorção de micronutrientes em virtude da redução da ingestão dietética e de sua associação com doença do intestino delgado ou pancreática. Outras causas de deficiências seriam: tratamento com drogas, peroxidação lipídica, déficit protéico, maior excreção urinária e aumento da necessidade e degradação de nutrientes. Como conseqüências dessas deficiências, esses pacientes apresentam usualmente anemia, esteatose hepática, estresse oxidativo e imunossupressão.

Maternal protein restriction affects gene expression and enzyme activity of intestinal disaccharidases in adult rat offspring

This study investigated the consequences of intrauterine protein restriction on the gastrointestinal tract and particularly on the gene expression and activity of intestinal disaccharidases in the adult offspring. Wistar rat dams were fed isocaloric diets containing 6% protein (restricted, n = 8) or 17% protein (control, n = 8) throughout gestation. Male offspring (n = 5-8 in each group) were evaluated at 3 or 16 weeks of age. Maternal protein restriction during pregnancy produced offspring with growth restriction from birth (5.7 ± 0.1 vs 6.3 ± 0.1 g; mean ± SE) to weaning (42.4 ± 1.3 vs 49.1 ± 1.6 g), although at 16 weeks of age their body weight was similar to control (421.7 ± 8.9 and 428.5 ± 8.5 g). Maternal protein restriction also increased lactase activity in the proximal (0.23 ± 0.02 vs 0.15 ± 0.02), medial (0.30 ± 0.06 vs 0.14 ± 0.01) and distal (0.43 ± 0.07 vs 0.07 ± 0.02 U·g-1·min-1) small intestine, and mRNA lactase abundance in the proximal intestine (7.96 ± 1.11 vs 2.38 ± 0.47 relative units) of 3-week-old offspring rats. In addition, maternal protein restriction increased sucrase activity (1.20 ± 0.02 vs 0.91 ± 0.02 U·g-1·min-1) and sucrase mRNA abundance (4.48 ± 0.51 vs 1.95 ± 0.17 relative units) in the duodenum of 16-week-old rats. In conclusion, the present study shows for the first time that intrauterine protein restriction affects gene expression of intestinal enzymes in offspring.