Padrão histológico, perfil imunoistoquímico e potencial pré-maligno das lesões diaplásicas da próstata canina

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Use of probiotics to reduce faecal shedding of Shiga toxin-producing Escherichia coli in sheep

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

The methylation patterns of the IGF2 and IGF2R genes in bovine spermatozoa are not affected by flow-cytometric sex sorting

The objectives of this study were to investigate the effect of sexing by flow cytometry on the methylation patterns of the IGF2 and IGF2R genes. Frozen-thawed, unsorted, and sex-sorted sperm samples from four Nellore bulls were used. Each ejaculate was separated into three fractions: non-sexed (NS), sexed for X-sperm (SX), and sexed for Y-sperm (SY). Sperm were isolated from the extender, cryoprotectant, and other cell types by centrifugation on a 40:70% Percoll gradient, and sperm pellets were used for genomic DNA isolation. DNA was used for analyses of the methylation patterns by bisulfite sequencing. Methylation status of the IGF2 and IGF2R genes were evaluated by sequencing 195 and 147 individual clones, respectively. No global differences in DNA methylation were found between NS, SX, and SY groups for the IGF2 (P=0.09) or IGF2R genes (P=0.38). Very specific methylation patterns were observed in the 25th and 26th CpG sites in the IGF2R gene. representing higher methylation in NS than in the SX and SY groups compared with the other CpG sites. Further, individual variation in methylation patterns was found among bulls. In conclusion, the sex-sorting procedure by flow cytometry did not affect the overall DNA methylation patterns of the IGF2 and IGF2R genes, although individual variation in their methylation patterns among bulls was observed. Mol. Reprod. Dev. 79:7784, 2012. (C) 2011 Wiley Periodicals, Inc.

Phenotypic and genotypic identification of streptococci and related bacteria isolated from bovine intramammary infections.

BACKGROUND Streptococcus spp. and other Gram-positive, catalase-negative cocci (PNC) form a large group of microorganisms which can be found in the milk of cows with intramammary infection. The most frequently observed PNC mastitis pathogens (major pathogens) are Streptococcus uberis, Strep. dysgalactiae, and Strep. agalactiae. The remaining PNC include a few minor pathogens and a large nonpathogenic group. Improved methods are needed for the accurate identification and differentiation of PNC. A total of 151 PNC were collected from cows with intramammary infection and conclusively identified by 16S rRNA sequencing as reference method. Nine phenotypic microbiological tests (alpha-hemolysis, CAMP reaction, esculin hydrolysis, growth on kanamycin esculin azide agar and on sodium chloride agar, inulin fermentation, hippurate hydrolysis, leucine aminopeptidase and pyrrolidonyl peptidase activity), multiplex PCR for the three major pathogens (target genes for Strep. uberis, Strep. dysgalactiae and Strep. agalactiae: pauA, 16S rRNA, and sklA3, respectively), and mass spectroscopy using the matrix-assisted laser desorption ionization-time of flight (MALDI-TOF MS) were evaluated for the diagnosis and discrimination of the three clinically most relevant PNC. RESULTS The probability that a strain of Strep. uberis, Strep. dysgalactiae and Strep. agalactiae was correctly identified by combining the results of the 9 phenotypic tests was 92%, 90%, and 100%, respectively. Applying the multiplex PCR, all strains of the three major pathogens were correctly identified and no false positive results occurred. Correct identification was observed for all strains of Strep. uberis and Strep. agalactiae using MALDI-TOF MS. In the case of Strep. dysgalactiae, some variability was observed at the subspecies level, but all strains were allocated to one single cluster. CONCLUSIONS The results of the present study show that reliable identification of the clinically most relevant PNC (Strep. uberis, Strep. agalactiae and Strep. dysgalactiae) can be obtained by use of a combination of colony morphology, hemolysis type and catalase reaction, and a multiplex PCR with specific primers restricted to these 3 pathogens. The MALDI-TOF MS is a fast method that shows promising results, although identification of Strep. dysgalactiae at the subspecies level is not yet satisfactory.

Characterization of the amp genes involved in the regulation of beta-lactamase expression in Pseudomonas aeruginosa

Antibiotic resistance, production of alginate and virulence factors, and altered host immune responses are the hallmarks of chronic Pseudomonas aeruginosa infection. Failure of antibiotic therapy has been attributed to the emergence of P. aeruginosa strains that produce β-lactamase constitutively. In Enterobacteriaceae, β-lactamase induction involves four genes with known functions: ampC, ampR, ampD, and ampG, encoding the enzyme, transcriptional regulator, amidase and permease, respectively. In addition to all these amp genes, P. aeruginosa possesses two ampG paralogs, designated ampG and ampP. In this study, P. aeruginosa ampC, ampR, ampG and ampP were analyzed. Inactivation of ampC in the prototypic PAO1 failed to abolish the β-lactamase activity leading to the discovery of P. aeruginosa oxacillinase PoxB. Cloning and expression of poxB in Escherichia coli confers β-lactam resistance. Both AmpC and PoxB contribute to P. aeruginosa resistance against a wide spectrum of β-lactam antibiotics. The expression of PoxB and AmpC is regulated by a LysR-type transcriptional regulator AmpR that up-regulates AmpC but down-regulates PoxB activities. Analyses of P. aeruginosa ampR mutant demonstrate that AmpR is a global regulator that modulates the expressions of Las and Rhl quorum sensing (QS) systems, and the production of pyocyanin, LasA protease and LasB elastase. Introduction of the ampR mutation into an alginate-producing strain reveals the presence of a complex co-regulatory network between antibiotic resistance, QS alginate and other virulence factor production. Using phoA and lacZ protein fusion analyses, AmpR, AmpG and AmpP were localized to the inner membrane with one, 16 and 10 transmembrane helices, respectively. AmpR has a cytoplasmic DNA-binding and a periplasmic substrate binding domains. AmpG and AmpP are essential for the maximal expression of β-lactamase. Analysis of the murein breakdown products suggests that AmpG exports UDP-N-acetylmuramyl-L-alanine-γ-D-glutamate-meso-diaminopimelic acid-D-alanine-D-alanine (UDP-MurNAc-pentapeptide), the corepressor of AmpR, whereas AmpP imports N-acetylglucosaminyl-beta-1,4-anhydro-N-acetylmuramic acid-Ala-γ-D-Glu-meso-diaminopimelic acid (GlcNAc-anhMurNAc-tripeptide) and GlcNAc-anhMurNAc-pentapeptide, the co-inducers of AmpR. This study reveals a complex interaction between the Amp proteins and murein breakdown products involved in P. aeruginosa β-lactamase induction. In summary, this dissertation takes us a little closer to understanding the P. aeruginosa complex co-regulatory mechanism in the development of β-lactam resistance and establishment of chronic infection. ^

Correlation Between The Friedman Staging System And The Upper Airway Volume In Patients With Obstructive Sleep Apnea.

This study was designed to evaluate the correlation between computed tomography findings and data from the physical examination and the Friedman Staging System (FSS) in patients with obstructive sleep apnea (OSA). We performed a retrospective evaluation by reviewing the medical records of 33 patients (19 male and 14 female patients) with a mean body mass index of 30.38 kg/m(2) and mean age of 49.35 years. Among these patients, 14 presented with severe OSA, 7 had moderate OSA, 7 had mild OSA, and 5 were healthy. The patients were divided into 2 groups according to the FSS: Group A comprised patients with FSS stage I or II, and group B comprised patients with FSS stage III. By use of the Fisher exact test, a positive relationship between the FSS stage and apnea-hypopnea index (P = .011) and between the FSS stage and body mass index (P = .012) was found. There was no correlation between age (P = .55) and gender (P = .53) with the FSS stage. The analysis of variance test comparing the upper airway volume between the 2 groups showed P = .018. In this sample the FSS and upper airway volume showed an inverse correlation and were useful in analyzing the mechanisms of airway collapse in patients with OSA.

Variação sazonal da fotossíntese, condutância estomática e potencial da água na folha de laranjeira 'Valência'

Seasonal variation in environmental conditions may influence gas exchange rates as well as water relations in perennial species. This work was carried out to evaluate photosynthetic rates (A), transpiration (E), stomatal conductance (g) and leaf water potential (psi f ) in 'Valencia' orange trees grafted on four different rootstocks. Measurements were made twice a day: from 9h00 to 11h00 a.m. and from 1h00 to 3h00 p.m., during January, March and July. A and g were significantly lower and psif was significantly more negative, in the afternoon. The decrease in A may be related to the reduction in g, due to the increase in the vapor pressure deficit between the air and the leaf (VPDair-leaf ) in the afternoon, when temperatures are higher. In spite of the partial stomatal closure in the afternoon, the values for E were approximately the same as those measured in the morning, due to the increase in the VPDair-leaf . A decrease in A and g could also be noted from January to July, that is, from the hot and humid summer months, to the colder and drier winter ones. It was suggested that the decrease in A and g observed from January through March, may be related to the decrease in plant growth rates, which could have influenced the source-sink relationships, since the climatic conditions for both months were similar. The decrease in A and g showed in July, seems to be related to the decrease in both the night temperature and the growth rate of plants.

Influência da expansão rápida da maxila na rizólise dos caninos decíduos usados como ancoragem

OBJETIVOS: o presente estudo radiográfico, retrospectivo e longitudinal objetivou determinar se a ancoragem do aparelho expansor fixo tipo Haas, modificado para as dentaduras decídua e mista, interfere na velocidade de rizólise e esfoliação dos caninos decíduos. MÉTODOS: foi feita uma avaliação quantitativa da rizólise do canino decíduo mediante a medição do comprimento coroa-ápice dos caninos decíduos superiores, dos lados direito e esquerdo. Para essa avaliação, 24 crianças submetidas à expansão rápida da maxila (ERM) na dentadura decídua ou no início da dentadura mista foram comparadas com 15 crianças que não passaram por esse procedimento. A medição do comprimento coroa-ápice dos caninos decíduos foi realizada com o programa computadorizado CEF-X Cefalometria Digital, produzido pela CDT Informática, que permitiu calibrar o tamanho dos dentes pela uniformização das imagens radiográficas digitalizadas. RESULTADOS: os dados estatísticos revelaram que não houve diferença na velocidade de rizólise dos caninos decíduos entre as crianças do grupo controle e as do grupo submetido à ERM. CONCLUSÕES: é possível inferir que o aparelho expansor fixo tipo Haas ancorado em dentes decíduos não influencia a rizólise dos caninos decíduos usados como ancoragem.

Influence of light and temperature on seed germination of Cereus pernambucensis Lemaire (Cactaceae)

The objective of the present study was to evaluate the effects of light and temperature on germination of Cereus pernambucensis seeds, a species of columnar cactus native to Brazil and naturally incident in the restinga. Cereus pernambucensis seeds were incubated under different temperatures, from 5 to 45 °C, with 5 °C intervals, and under alternating temperatures of 15-20 °C, 15-30 °C, 20-25 °C, 20-30 °C, 20-35 °C, 25-30 °C, 25-35 °C, and 30-35 °C, both under continuous white light and dark. The seeds were also incubated in a gradient of phytochrome photoequilibrium at 25 °C. The highest percentage germination in this species was between 25 and 30 °C. The minimum temperature was between 15 and 20 °C and the maximum between 35 and 40 °C. Alternating temperatures did not affect the percentage of seed germination, but it did alter the rate and synchronization indexes. Seeds incubated in the dark did not germinate under any of the conditions tested, indicating that this species when cultivated present light sensitive seeds controlled by phytochrome. The seeds can tolerate a lot of shade conditions, germinating under very low fluence response of phytochrome.

Relações entre conteúdo e forma de conhecimentos e práticas pedagógicas em Geociências: imaginário de futuros professores numa disciplina de licenciatura

Pesquisas e reflexões sobre ensino, história e epistemologia das Ciências da Terra têm sido desenvolvidas, em grande parte, intimamente associadas às práticas docentes dos professores/pesquisadores universitários. Nessa mesma perspectiva, buscamos neste trabalho compreender relações que os estudantes de licenciatura em Geografia estabelecem entre o conteúdo/forma trabalhado numa disciplina universitária e o conteúdo/forma profissional, ou seja, pensado na relação com a escola básica, com o tornar-se professor. As respostas dos estudantes a uma questão por nós formulada indicou como eles representaram uma das atividades desenvolvidas na disciplina na relação com seu imaginário sobre ser professor da educação básica. As análises mostraram também que essas significações se produzem de maneiras muito diferentes entre os estudantes, fazendo intervir no imaginário a partir do qual essas significações se produzem, aspectos da memória escolar de alguns estudantes e da maneira como representam o conhecimento escolar e as práticas de ensino na disciplina Geografia da educação básica. As análises mostraram que a atividade não é transparente quanto ao que seja o "conteúdo" de ensino da atividade, posto que essas significações são o trabalho de diferentes memórias discursivas, implicado em diferentes posições-aluno. Este estudo aponta para a importância de considerar e para a possibilidade de trabalhar com os imaginários dos futuros professores sobre ensino na relação que estabelecem com os conteúdos/formas em suas disciplinas científicas universitárias.

Brazilian offshore wave climate based on NWW3 reanalysis

This paper provides a description of the wave climate off the Brazilian coast based on an eleven-year time series (Jan/1997-Dec/2007) obtained from the NWW3 operational model hindcast reanalysis. Information about wave climate in Brazilian waters is very scarce and mainly based on occasional short-term observations, the present analysis being the first covering such temporal and spatial scales. To define the wave climate, six sectors were defined and analyzed along the Brazilian shelf-break: South (W1), Southeast (W2), Central (W3), East (W4), Northeast (W5) and North (W6). W1, W2 and W3 wave regimes are determined by the South Atlantic High (SAH) and the passage of synoptic cold fronts; W4, W5 and W6 are controlled by the Intertropical Convergence Zone (ITCZ) and its meridional oscillation. The most energetic waves are from the S, generated by the strong winds associated to the passage of cold fronts, which mainly affect the southern region. Wave power presents a decrease in energy levels from south to north, with its annual variation showing that the winter months are the most energetic in W1 to W4, while in W5 and W6 the most energetic conditions occur during the austral summer. The information presented here provides boundary conditions for studies related to coastal processes, fundamental for a better understanding of the Brazilian coastal zone.

Georges Perec: o anti-proust?

This article presents and analyses the points of view of literary critics that have studied Marcel Proust's role in the work of Georges Perec with the intention of showing the workings of their approaches. The article also points towards a direction that can be followed when analyzing these two writers' relations.

Amazon deforestation alters small stream structure, nitrogen biogeochemistry and connectivity to larger rivers

Human activities that modify land cover can alter the structure and biogeochemistry of small streams but these effects are poorly known over large regions of the humid tropics where rates of forest clearing are high. We examined how conversion of Amazon lowland tropical forest to cattle pasture influenced the physical and chemical structure, organic matter stocks and N cycling of small streams. We combined a regional ground survey of small streams with an intensive study of nutrient cycling using (15)N additions in three representative streams: a second-order forest stream, a second-order pasture stream and a third-order pasture stream. These three streams were within several km of each other and on similar soils. Replacement of forest with pasture decreased stream habitat complexity by changing streams from run and pool channels with forest leaf detritus (50% cover) to grass-filled (63% cover) channel with runs of slow-moving water. In the survey, pasture streams consistently had lower concentrations of dissolved oxygen and nitrate (NO(3) (-)) compared with similar-sized forest streams. Stable isotope additions revealed that second-order pasture stream had a shorter NH(4) (+) uptake length, higher uptake rates into organic matter components and a shorter (15)NH(4) (+) residence time than the second-order forest stream or the third-order pasture stream. Nitrification was significant in the forest stream (19% of the added (15)NH(4) (+)) but not in the second-order pasture (0%) or third-order (6%) pasture stream. The forest stream retained 7% of added (15)N in organic matter compartments and exported 53% ((15)NH(4) (+) = 34%; (15)NO(3) (-) = 19%). In contrast, the second-order pasture stream retained 75% of added (15)N, predominantly in grasses (69%) and exported only 4% as (15)NH(4) (+). The fate of tracer (15)N in the third-order pasture stream more closely resembled that in the forest stream, with 5% of added N retained and 26% exported ((15)NH(4) (+) = 9%; (15)NO(3) (-) = 6%). These findings indicate that the widespread infilling by grass in small streams in areas deforested for pasture greatly increases the retention of inorganic N in the first- and second-order streams, which make up roughly three-fourths of total stream channel length in Amazon basin watersheds. The importance of this phenomenon and its effect on N transport to larger rivers across the larger areas of the Amazon Basin will depend on better evaluation of both the extent and the scale at which stream infilling by grass occurs, but our analysis suggests the phenomenon is widespread.

A flow-based procedure with solenoid micro-pumps for the spectrophotometric determination of uric acid in urine

The determination of uric acid in urine shows clinical importance, once it can be related to human organism dysfunctions, such as gout. An analytical procedure employing a multicommuted flow system was developed for the determination of uric acid in urine samples. Cu(II) ions are reduced by uric acid to Cu(I) that can be quantified by spectrophotometry in the presence of 2,2`-biquinoline 4,4`-dicarboxylic acid (BCA). The analytical response was linear between 10 and 100 mu mol L(-1) uric acid with a detection limit of 3.0 mu mol L(-1) (99.7% confidence level). Coefficient of variation of 1.2% and sampling rate of 150 determinations per hour were achieved. Per determination, 32 mu g of CuSO(4) and 200 mu g of BCA were consumed, generating 2.0 mL of waste. Recoveries from 91 to 112% were estimated and the results for 7 urine samples agreed with those obtained by the commercially available enzymatic kit for determination of uric acid. The procedure required 100-fold dilution of urine samples, minimizing sample consumption and interfering effects. In order to avoid the manual dilution step, on-line sample dilution was achieved by a simple system reconfiguration attaining a sampling rate of 95 h(-1). (C) 2009 Elsevier B.V. All rights reserved.

Skewness, splitting number and vertex deletion of some toroidal meshes

The skewness sk(G) of a graph G = (V, E) is the smallest integer sk(G) >= 0 such that a planar graph can be obtained from G by the removal of sk(C) edges. The splitting number sp(G) of C is the smallest integer sp(G) >= 0 such that a planar graph can be obtained from G by sp(G) vertex splitting operations. The vertex deletion vd(G) of G is the smallest integer vd(G) >= 0 such that a planar graph can be obtained from G by the removal of vd(G) vertices. Regular toroidal meshes are popular topologies for the connection networks of SIMD parallel machines. The best known of these meshes is the rectangular toroidal mesh C(m) x C(n) for which is known the skewness, the splitting number and the vertex deletion. In this work we consider two related families: a triangulation Tc(m) x c(n) of C(m) x C(n) in the torus, and an hexagonal mesh Hc(m) x c(n), the dual of Tc(m) x c(n) in the torus. It is established that sp(Tc(m) x c(n)) = vd(Tc(m) x c(n) = sk(Hc(m) x c(n)) = sp(Hc(m) x c(n)) = vd(Hc(m) x c(n)) = min{m, n} and that sk(Tc(m) x c(n)) = 2 min {m, n}.