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101.
A ribosomal RNA gene intergenic spacer based PCR and DGGE fingerprinting method for the analysis of specific rhizobial communities in soil 总被引:5,自引:0,他引:5
de Oliveira VM Manfio GP da Costa Coutinho HL Keijzer-Wolters AC van Elsas JD 《Journal of microbiological methods》2006,64(3):366-379
A direct molecular method for assessing the diversity of specific populations of rhizobia in soil, based on nested PCR amplification of 16S-23S ribosomal RNA gene (rDNA) intergenic spacer (IGS) sequences, was developed. Initial generic amplification of bacterial rDNA IGS sequences from soil DNA was followed by specific amplification of (1) sequences affiliated with Rhizobium leguminosarum "sensu lato" and (2) R. tropici. Using analysis of the amplified sequences in clone libraries obtained on the basis of soil DNA, this two-sided method was shown to be very specific for rhizobial subpopulations in soil. It was then further validated as a direct fingerprinting tool of the target rhizobia based on denaturing gradient gel electrophoresis (DGGE). The PCR-DGGE approach was applied to soils from fields in Brazil cultivated with common bean (Phaseolus vulgaris) under conventional or no-tillage practices. The community fingerprints obtained allowed the direct analysis of the respective rhizobial community structures in soil samples from the two contrasting agricultural practices. Data obtained with both primer sets revealed clustering of the community structures of the target rhizobial types along treatment. Moreover, the DGGE profiles obtained with the R. tropici primer set indicated that the abundance and diversity of these organisms were favoured under NT practices. These results suggest that the R. leguminosarum-as well as R. tropici-targeted IGS-based nested PCR and DGGE are useful tools for monitoring the effect of agricultural practices on these and related rhizobial subpopulations in soils. 相似文献
102.
103.
Ítalo Antônio Cotta Coutinho Dayana Maria Teodoro FrancinoAristéa Alves Azevedo Renata Maria Strozi Alves Meira 《Flora》2012
In this paper the ontogenesis and histochemistry of the petiolar glands found on the petiole/rachis of the eight Chamaecrista species of the section Absus, subsection Baseophyllum (Leguminosae, Caesalpinioideae) are studied by using light microscopy techniques, aiming to characterise these structures and to provide taxonomic characters which may be useful in phylogenetic approaches. Strips for glucose identification reacted positively with the exudates of the glands, confirming the presence of nectar in the secretion, characterising these glands as extrafloral nectaries (EFN). Histochemical tests also detected the presence of neutral and acid muco-polysaccharides, pectins, mucilages, total proteins, and phenolic compounds in the EFNs. The EFNs arise from a group of meristem cells (protodermis, ground meristem and procambium) in the petiole/rachis. All EFNs of the investigated taxa share some morpho-anatomical characters, so that their peculiarities are too weak to be used alone in the identification of particular species. Rather their similarities may be used to include these species into a single group, supporting the hypothesis of monophyly of the subsection Baseophyllum. 相似文献
104.
Santos KK Matias EF Tintino SR Souza CE Braga MF Guedes GM Rolón M Vega C de Arias AR Costa JG Menezes IR Coutinho HD 《Experimental parasitology》2012,131(1):130-132
Chagas disease is caused by Trypanosoma cruzi, being considered a public health problem. An alternative to combat this pathogen is the use of natural products isolated from fruits such as Eugenia uniflora, a plant used by traditional communities as food and medicine due to its antimicrobial and biological activities. Ethanolic extract from E. uniflora was used to evaluate in vitro anti-epimastigote and cytotoxic activity. This is the first record of anti-Trypanosoma activity of E. uniflora, demonstrating that a concentration presenting 50% of activity (EC(50)) was 62.76 μg/mL. Minimum inhibitory concentration (MIC) was ≤ 1024 μg/mL. Our results indicate that E. uniflora could be a source of plant-derived natural products with anti-epimastigote activity with low toxicity. 相似文献
105.
Nones K Ledur MC Zanella EL Klein C Pinto LF Moura AS Ruy DC Baron EE Ambo M Campos RL Boschiero C Burt DW Coutinho LL 《Animal genetics》2012,43(5):570-576
Major objectives of the poultry industry are to increase meat production and to reduce carcass fatness, mainly abdominal fat. Information on growth performance and carcass composition are important for the selection of leaner meat chickens. To enhance our understanding of the genetic architecture underlying the chemical composition of chicken carcasses, an F2 population developed from a broiler × layer cross was used to map quantitative trait loci (QTL) affecting protein, fat, water and ash contents in chicken carcasses. Two genetic models were applied in the QTL analysis: the line‐cross and the half‐sib models, both using the regression interval mapping method. Six significant and five suggestive QTL were mapped in the line‐cross analysis, and four significant and six suggestive QTL were mapped in the half‐sib analysis. A total of eleven QTL were mapped for fat (ether extract), five for protein, four for ash and one for water contents in the carcass using both analyses. No study to date has reported QTL for carcass chemical composition in chickens. Some QTL mapped here for carcass fat content match, as expected, QTL regions previously associated with abdominal fat in the same or in different populations, and novel QTL for protein, ash and water contents in the carcass are presented here. The results described here also reinforce the need for fine mapping and to perform multi‐trait analyses to better understand the genetic architecture of these traits. 相似文献
106.
Campos LB Pucca MB Roncolato EC Netto JC Barbosa JE 《Journal of biochemical and molecular toxicology》2012,26(8):308-314
The study of venom components is an important step toward understanding the mechanism of action of such venoms and is indispensable for the development of new therapies. This work aimed to investigate the venom of Lachesis muta rhombeata and evaluate enzymes related to its toxicity. Phospholipase A2 (PLA2), l ‐amino acid oxidase (LAAO), and proteinase activities were measured, and the molecular weights were estimated. We found the venom to contain one PLA2 (17 kDa), one LAAO (132 kDa), and three serine proteinases (40, 31, and 20 kDa). Although only serine proteinases were observed in the zymogram, metalloproteinases were found to contribute more to the total proteolytic activity than did serine proteinases. The work confirmed the presence of highly active enzymes; and, moreover, we proposed a novel method for confirming the presence of LAAOs by zymography. We also suggested a simple step to increase the sensitivity of proteinase assays. © 2012 Wiley Periodicals, Inc. J Biochem Mol Toxicol 26:308–314, 2012; View this article online at wileyonlinelibrary.com. DOI 10:1002/jbt.21422 相似文献
107.
Brad J. Sagarin Amy L. Martin Savia A. Coutinho John E. Edlund Lily Patel John J. Skowronski Bettina Zengel 《Evolution and human behavior》2012,33(6):595-614
The theory of evolved sex differences in jealousy predicts sex differences in responses to sexual infidelities and emotional infidelities. Critics have argued that such differences are absent in studies that use continuous measures to assess responses to hypothetical infidelities or in studies that assess responses to real infidelities. These criticisms were tested in two random-effects meta-analyses of 40 published and unpublished papers (providing 209 effect sizes from 47 independent samples) that measured sex differences in jealousy using continuous measures. A significant, theory-supportive sex difference emerged across 45 independent samples using continuous measures of responses to hypothetical infidelities, g*=0.258, 95% confidence interval (CI) [0.188, 0.328], p<.00001. Measured emotion significantly moderated effect size. Effects were strongest when measures assessed distress/upset (g*=0.337) and jealousy (g*=0.309). Other commonly measured negative emotions yielded weaker effects, including hurt (g*=0.161), anger (g*=0.074), and disgust (g*=0.012). Across the 45 independent samples, six significant moderators emerged: random sampling, population type (student vs. nonstudent samples), age, inclusion of a forced-choice question, number of points in the response scale, and year of publication. A significant, theory-supportive effect also emerged across seven studies assessing reactions to actual infidelities, g*=0.234, 95% CI [0.020, 0.448], p=.03. Results demonstrate that the sex difference in jealousy neither is an artifact of response format nor is limited to responses to hypothetical infidelities. 相似文献
108.
109.
Kroll-Palhares K Silvério JC Silva AA Michailowsky V Marino AP Silva NM Carvalho CM Pinto LM Gazzinelli RT Lannes-Vieira J 《Memórias do Instituto Oswaldo Cruz》2008,103(4):375-385
In Chagas disease, understanding how the immune response controls parasite growth but also leads to heart damage may provide insight into the design of new therapeutic strategies. Tumor necrosis factor-alpha (TNF-alpha) is important for resistance to acute Trypanosoma cruzi infection; however, in patients suffering from chronic T. cruzi infection, plasma TNF-alpha levels correlate with cardiomyopathy. Recent data suggest that CD8-enriched chagasic myocarditis formation involves CCR1/CCR5-mediated cell migration. Herein, the contribution of TNF-alpha, especially signaling through the receptor TNFR1/p55, to the pathophysiology of T. cruzi infection was evaluated with a focus on the development of myocarditis and heart dysfunction. Colombian strain-infected C57BL/6 mice had increased frequencies of TNFR1/p55+ and TNF-alpha+ splenocytes. Although TNFR1-/- mice exhibited reduced myocarditis in the absence of parasite burden, they succumbed to acute infection. Similar to C57BL/6 mice, Benznidazole-treated TNFR1-/- mice survived acute infection. In TNFR1-/- mice, reduced CD8-enriched myocarditis was associated with defective activation of CD44+CD62Llow/- and CCR5+ CD8+ lymphocytes. Also, anti-TNF-alpha treatment reduced the frequency of CD8+CCR5+ circulating cells and myocarditis, though parasite load was unaltered in infected C3H/HeJ mice. TNFR1-/- and anti-TNF-alpha-treated infected mice showed regular expression of connexin-43 and reduced fibronectin deposition, respectively. Furthermore, anti-TNF-alpha treatment resulted in lower levels of CK-MB, a cardiomyocyte lesion marker. Our results suggest that TNF/TNFR1 signaling promotes CD8-enriched myocarditis formation and heart tissue damage, implicating the TNF/TNFR1 signaling pathway as a potential therapeutic target for control of T. cruzi-elicited cardiomyopathy. 相似文献
110.
Cuervo P Cupolillo E Britto C González LJ E Silva-Filho FC Lopes LC Domont GB De Jesus JB 《Journal of Proteomics》2008,71(1):109-122
A comparative analysis of proteomic maps of long-term grown and fresh clinical Trichomonas vaginalis isolates exhibiting low and high virulence phenotypes, respectively, was performed using two-dimensional gel electrophoresis and mass spectrometry. Of 29 protein spots differentially expressed between the isolates, 19 were over-expressed in the isolate exhibiting high virulence phenotype: proteins associated with cytoskeletal dynamics, such as coronin and several isoforms of actin, as well as proteins involved in signal transduction, protein turnover, proteolysis, and energetic and polyamine metabolisms were identified. Some malate dehydrogenase, fructose-1,6-bisphosphate aldolase and ornithine cyclodeamidase isoforms were exclusively expressed by the highly virulent isolate. During interaction assays with VEC, parasites exhibiting high virulence phenotype rapidly adhered and switched to amoeboid forms. In contrast, low adhesion and no morphological transformation were observed in parasites displaying low virulence phenotype. Our findings demonstrate that expression of specific proteins by high and low virulence parasites could be associated with the ability of each isolate to undergo morphological transformation and interact with host cells. Such data represent an important step towards understanding of the complex interaction network of proteins that participate in the mechanism of pathogenesis of this protozoan. 相似文献