Electrochemical genosensors for the detection of Bonamia parasite. Selection of single strand-DNA (ssDNA) probes by simulation of the secondary structure folding

A post-PCR nucleic acid work by comparing experimental data, from electrochemical genosensors, and bioinformatics data, derived from the simulation of the secondary structure folding and prediction of hybridisation reaction, was carried out in order to rationalize the selection of ssDNA probes for the detection of two Bonamia species, B. exitiosa and B. ostreae, parasites of Ostrea edulis.Six ssDNA probes (from 11 to 25 bases in length, 2 thiolated and 4 biotinylated) were selected within different regions of B. ostreae and B. exitiosa PCR amplicons (300 and 304 bases, respectively) with the aim to discriminate between these parasite species. ssDNA amplicons and probes were analyzed separately using the "Mfold Web Server" simulating the secondary structure folding behaviour. The hybridisation of amplicon-probe was predicted by means of "Dinamelt Web Server". The results were evaluated considering the number of hydrogen bonds broken and formed in the simulated folding and hybridisation process, variance in gaps for each sequence and number of available bases. In the experimental part, thermally denatured PCR products were captured at the sensor interface via sandwich hybridisation with surface-tethered probes (thiolated probes) and biotinylated signalling probes. A convergence between analytical signals and simulated results was observed, indicating the possibility to use bioinformatic data for ssDNA probes selection to be incorporated in genosensors. (C) 2011 Elsevier B.V. All rights reserved.

Tonometria, paquimetria e comprimento axial ocular em cães glaucomatosos submetidos à ablação uveal intravítrea

Estudaram-se as alterações tonométricas, paquimétricas e de comprimento axial em cães com glaucoma submetidos à ablação uveal intravítrea. Foram avaliados 13 olhos irreversivelmente cegos de cães que apresentavam glaucoma crônico unilateral, nos quais realizou-se a ablação uveal intra-vítrea, por meio de injeção na câmara vítrea de 0,5ml de sulfato de gentamicina (40mg/ml) associado a 0,3ml de fosfato de dexametasona (4mg/ml). As mensurações da pressão intra-ocular (Po), espessura corneana e eixo axial com tonometria de aplanação, paquimetria ultra-sônica e ultra-sonografias modos A e B foram realizadas no dia da ablação (M0) e após uma (M1), quatro (M4), oito (M8), 12 (M12), 24 (M24) e 48 semanas (M48). A Po diminuiu significativamente em todos os momentos em relação ao M0, com aumento da espessura corneana ao longo do experimento. Nas avaliações ultra-sonográficas notou-se diminuição significativa do bulbo ocular a partir de M4, com medidas ultra-sonográficas modo A significativamente maiores que as do modo B. O procedimento foi efetivo na redução da Po e na diminuição do eixo axial, demonstrando ser viável no controle do glaucoma crônico em olhos irreversivelmente cegos, e ser uma alternativa à enucleação e inserção de prótese ocular.

Transplante de membrana amniótica canina criopreservada para cicatrização de córnea com deficiência de células límbicas em coelhos

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

Relation between photoluminescence emission and local order-disorder in the CaTiO3 lattice modifier

In this letter, the authors propose that photoluminescence emission in CaTiO3 is affected not only by disorder in the lattice former but also by structural disorder in the lattice modifier. Structural disorder was evaluated by Ti, Ca K-edge x-ray absorption near-edge structure experiments and by photoluminescence emission. The preedge feature of the Ca K edge was related to the intensity of photoluminescence emission. The results of the preedge feature of the Ca K-edge x-ray absorption near-edge structure confirm the presence of different Ca coordination numbers, namely, Ca-O-11 and Ca-O-12. (c) 2007 American Institute of Physics.

Characterization of electrodes chemically modified with Mn(III) porphyrin/polypyrrole films as catalytic surfaces for an azo dye

In this study we describe the electrochemical behavior of 5,10,15,20-tetrakis(2'-aminophenylporphyrin)manganese(III) chloride supported on a glassy carbon electrode, as well as the electrochemical preparation and characterization of thin films based on pyrrole-3-carboxylic acid. The electrocatalytic action of the electrode modified with the Mn(III) porphyrin toward an azo dye was tested, and the characteristic strong interaction between the incorporated metalloporphyrin and RR120 dye was verified. Copyright (c) 2006 Society of Porphyrins & Phthalocyanines.

Experimental alcoholism and pathogenesis of prostatic diseases in UChB rats

Previous studies have shown that long-term alcohol treatment has negative effects on prostatic stromal-epithelial interaction. Thus, the aim of the present study was to analyze the histochemical, immunohistochemical and ultrastructural alterations that occur in the prostatic stroma and epithelium of rats submitted to chronic alcohol ingestion and alcohol abstinence, as well as to establish the relationship between these changes and prostatic diseases. Thirty male rats (10 Wistar and 20 UChB rats) were divided into three experimental groups: the control group received tap water, the alcoholic group received ethanol diluted to 10 degrees G.L. for 150 days, and the abstinent group received the same liquid diet as the alcoholic group up to 120 days of treatment and only tap water for 30 days thereafter. At the end of treatment, all animals were sacrificed and the ventral lobe of the prostate was removed and processed for histochemical, immunohistochemical and ultrastructural analyses. In addition, plasma testosterone levels were measured. The results showed, prostatic intraepithelial neoplasia, infolding of the epithelium towards the stroma, stromal hypertrophy and the presence of inflammatory cells in alcoholic animals. In the abstinent group, alterations were noted mainly in the stromal area. In conclusion, ethanol triggers alterations in prostatic epithelial and stromal compartments, affecting the stromal microenvironment and predisposing the organ to pathological processes. (C) 2006 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.

Atividade antimicrobiana de Lactobacillus acidophilus, contra microrganismos patogênicos veiculados por alimentos

The antimicrobial activity of a commercial probiotic culture, Lactobacillus acidophilus (La5), was tested against two foodborne pathogens, Escherichia coli and Staphylococcus aureus. The antagonistic effect of the probiotic culture in vitro was performed by applying both Multilayer Agar Plate and Agar Well Diffusion methods. The results indicated that the inhibitory substance present on 72 hours culture broth supernatant was extracellular and diffusible. The incubation period of the lactic acid bacteria on MRS Broth, at 3 7 degrees C in aerobic conditions, for the highest lactic acid production (1,08 g/%) was 72 hours, which gave a minimum pH value of the supernatant (3,90) and the best inhibition results by the Well Diffusion Agar Assay, showing inhibition zone diameters of 14,75mm and 15,0mm for E. coli and S. aureus, respectively. The inhibitor compound was not sensitive to proteolytic enzyme and freezing, but was totally inactivated when the supernatant was neutralized with NaOH 1 N solution. The results suggest that the inhibitory activity was due to the lactic acid concentration and the low pH of the probiotic culture broth.

Blue-green and red photoluminescence in CaTiO3 : Sm

Blue-green and red photoluminescence (PL) emission in structurally disordered CaTiO3:Sm (CT:Sm) powders was observed at room temperature with laser excitation at 350.7 nm. The perovskite-like titanate CT:Sm powders prepared by a soft chemical processing at different temperatures of annealing were structurally characterized by X-ray diffraction (XRD) and X-ray absorption near-edge structure (XANES). The results indicate that the generation of the broad PL band is related to order-disorder degree in the perovskitelike structure. (c) 2006 Elsevier B.V. All rights reserved.

Evaluation of transformation growth factor beta(1), interleukin-10, and interferon-gamma in male symptomatic and asymptomatic dogs naturally infected by Leishmania (Leishmania) chagasi

The aims of this study were to evaluate the immunomodulatory role of TGF-beta(1), 1L-10, and INF-gamma in spleen and liver extracts and supernatant cultures of white spleen cells from male symptomatic and asymptomatic dogs, naturally infected by Leishmania (Leishmania) chagasi. Thirty dogs from Aracatuba, São Paulo, Brazil, an endemic leishmaniosis area, were selected by positive ELISA serological reaction for Leishmania sp. and divided into two groups: asymptomatic (n=15) and symptomatic (n=15) consisting of animals with at least three characteristic signs (fever, dermatitis, lymphoadenopathy, onychogryphosis, weight loss, cachex a, locomotion problems, conjunctivitis, epistaxis, hepatosplenomegaly, edema, and apathy). After euthanasia, spleen and liver fragments were collected for ex vivo quantification of TGF-beta(1), IL-10, and INF-gamma. Naturally active in vitro produced TGF-beta(1) was also evaluated in spleen cell culture supernatant. Spleen and liver extract of asymptomatic dogs had higher mean TGF-beta(1) levels than symptomatic dogs. High concentrations of IL-10 were found in spleen, and mainly in liver extract of both groups. Higher INF-gamma concentrations were found in spleen extracts of symptomatic dogs, and in liver extracts of asymptomatic dogs. Extract of this cytokire was lower in spleen extract. Although INF-gamma is being produced in canine infection, mean levels of TGF-beta(1) and IL-10 from spleen and liver extracts were quantitatively much higher; suggesting that immune response in both asymptomatic and symptomatic dogs A as predominantly type Th2. (c) 2006 Elsevier B.V. All rights reserved.

Differential gene expression by Paracoccidioides brasiliensis in host interaction conditions: Representational difference analysis identifies candidate genes associated with fungal pathogenesis

Paracoccidioides brasiliensis causes infection by the host inhalation of airborne propagules of the mycelia phase of the fungus. These particles reach the lungs, and disseminate to virtually all organs. Here we describe the identification of differentially expressed genes in studies of host-fungus interaction. We analyzed two cDNA populations of P. brasiliensis, one obtained from infected animals and the other an admixture of fungus and human blood thus mimicking the hematologic events of the fungal dissemination. Our analysis identified transcripts differentially expressed. Genes related to iron acquisition, melanin synthesis and cell defense were specially upregulated in the mouse model of infection. The upregulated transcripts of yeast cells during incubation with human blood were those predominantly related to cell wall remodeling/synthesis. The expression pattern of genes was independently confirmed in host conditions, revealing their potential role in the infection process. This work can facilitate functional studies of novel regulated genes that may be important for the survival and growth strategies of P. brasiliensis in humans. (c) 2006 Elsevier Masson SAS. All rights reserved.

MOLECULAR DIAGNOSIS of LEISHMANIA AMAZONENSIS IN A CAPTIVE SPIDER MONKEY IN BAURU, São Paulo, BRAZIL

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

Seroepidemiology of Toxoplasma gondii, Neospora caninum, and Leishmania spp. infections and risk factors for cats from Brazil

The seroprevalence of infection by Toxoplasma gondii, Neospora caninum, and Leishmania spp. was detected through an indirect immunofluorescence in 70 cats from the Andradina Municipality, São Paulo State, Brazil. Anti-T. gondii antibodies (titer >64) were detected in 15.7% (11/70) of animals, whereas positivity for N. caninum (titer 16) was not observed in any animal. of the cats from urban and rural areas, 10.4% (5/48) and 27.2% (6/22) were positive for T. gondii, respectively. Breed, age, food, and contact with animals of other species were significant for considering the positivity for T. gondii (P <= 0.0001). Cats having access to streets (17.1%, 11/64), cats cohabiting with rats (19.6%, 10/51), and cats feeding on homemade food and raw milk (27.2%, 6/22) were positive for T. gondii. In addition, 4.2% (3/70) of the cats were positive for Leishmania spp. by ELISA technique and negative by IFAT without coinfection with T. gondii and Leishmania spp. There was no serological positivity against feline immunodeficiency virus or feline leukemia virus. In conclusion, T. gondii infection in part of the feline population from Andradina is not linked to immunosuppressions or coinfections but probably to postnatal infection in association with the type of diet and presence of rats.

Genetic diversity of Stryphnodendron adstringens (Mart.) Coville determined by AFLP molecular markers

Bark extracts of Stryphnodendron adstringens (Mart) Coville a Leguminosae species, well known in Brazil as barbatimao, are popularly used as healing agent. The objective of this work was to determine the genetic diversity of S. adstringens populations and to correlate genetic distances to the production of tannins. S. adstringens accessions from populations found in Cerrado regions in the states of Goias, Minas Gerais and São Paulo were analyzed using the AFLP (Amplified Fragment Length Polymorphism) technique. A total of 236 polymorphic bands were scored and higher proportion of genetic diversity was found inter populations (70.9%), rather than intra populations (29.1%). F-ST value was found to be significantly greater than zero (0.2906), demonstrating the complex genetic structure of S. adstringens populations. Accessions collected in Cristalina, GO, showed higher percentage of polymorphic loci (87.3%) and the highest genetic diversity. The lowest genetic variability was detected among accessions from the population growing in Caldas Novas, GO. The genetic distance among populations was estimated using the Unweighted Pair Group Method with Arithmetic Mean (UPGMA), which grouped populations into 3 clusters. Moreover, chemotypes with tannin concentration above 40% showed higher genetic similarity. AFLP analysis proved to be an efficient gene mapping technique to determine the genetic diversity among remaining populations of S. adstringens. Obtained results may be employed to implement further strategies for the conservation of this medicinal plant. (C) 2011 Elsevier Ltd. All rights reserved.

Hierarchical Assembly of CaMoO4 Nano-Octahedrons and Their Photoluminescence Properties

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)