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101.
Adriano Mondini Roberta Vieira de Moraes Bronzoni Silvia Helena Pereira Nunes Francisco Chiaravalloti Neto Eduardo Massad Wladimir J. Alonso Eduardo S. M. Lázzaro Amena Alcantara Ferraz Paolo Marinho de Andrade Zanotto Maurício Lacerda Nogueira 《PLoS neglected tropical diseases》2009,3(5)
The dengue virus has a single-stranded positive-sense RNA genome of ∼10.700 nucleotides with a single open reading frame that encodes three structural (C, prM, and E) and seven nonstructural (NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5) proteins. It possesses four antigenically distinct serotypes (DENV 1–4). Many phylogenetic studies address particularities of the different serotypes using convenience samples that are not conducive to a spatio-temporal analysis in a single urban setting. We describe the pattern of spread of distinct lineages of DENV-3 circulating in São José do Rio Preto, Brazil, during 2006. Blood samples from patients presenting dengue-like symptoms were collected for DENV testing. We performed M-N-PCR using primers based on NS5 for virus detection and identification. The fragments were purified from PCR mixtures and sequenced. The positive dengue cases were geo-coded. To type the sequenced samples, 52 reference sequences were aligned. The dataset generated was used for iterative phylogenetic reconstruction with the maximum likelihood criterion. The best demographic model, the rate of growth, rate of evolutionary change, and Time to Most Recent Common Ancestor (TMRCA) were estimated. The basic reproductive rate during the epidemics was estimated. We obtained sequences from 82 patients among 174 blood samples. We were able to geo-code 46 sequences. The alignment generated a 399-nucleotide-long dataset with 134 taxa. The phylogenetic analysis indicated that all samples were of DENV-3 and related to strains circulating on the isle of Martinique in 2000–2001. Sixty DENV-3 from São José do Rio Preto formed a monophyletic group (lineage 1), closely related to the remaining 22 isolates (lineage 2). We assumed that these lineages appeared before 2006 in different occasions. By transforming the inferred exponential growth rates into the basic reproductive rate, we obtained values for lineage 1 of R0 = 1.53 and values for lineage 2 of R0 = 1.13. Under the exponential model, TMRCA of lineage 1 dated 1 year and lineage 2 dated 3.4 years before the last sampling. The possibility of inferring the spatio-temporal dynamics from genetic data has been generally little explored, and it may shed light on DENV circulation. The use of both geographic and temporally structured phylogenetic data provided a detailed view on the spread of at least two dengue viral strains in a populated urban area. 相似文献
102.
In China, croplands account for a relatively large form of vegetation cover. Quantifying carbon dioxide exchange and understanding the environmental controls on carbon fluxes over croplands are critical in understanding regional carbon budgets and ecosystem behaviors. In this study, the net ecosystem exchange (NEE) at a winter wheat/summer maize rotation cropping site, representative of the main cropping system in the North China Plain, was continuously measured using the eddy covariance technique from 2005 to 2009. In order to interpret the abiotic factors regulating NEE, NEE was partitioned into gross primary production (GPP) and ecosystem respiration (Reco). Daytime Reco was extrapolated from the relationship between nighttime NEE and soil temperature under high turbulent conditions. GPP was then estimated by subtracting daytime NEE from the daytime estimates of Reco. Results show that the seasonal patterns of the temperature responses of Reco and light‐response parameters are closely related to the crop phenology. Daily Reco was highly dependent on both daily GPP and air temperature. Interannual variability showed that GPP and Reco were mainly controlled by temperature. Water availability also exerted a limit on Reco. The annual NEE was ?585 and ?533 g C m?2 for two seasons of 2006–2007 and 2007–2008, respectively, and the wheat field absorbed more carbon than the maize field. Thus, we concluded that this cropland was a strong carbon sink. However, when the grain harvest was taken into account, the wheat field was diminished into a weak carbon sink, whereas the maize field was converted into a weak carbon source. The observations showed that severe drought occurring during winter did not reduce wheat yield (or integrated NEE) when sufficient irrigation was carried out during spring. 相似文献
103.
RACE技术是一项扩增基因末端序列的新技术。该研究从牛BMP4基因出发,以牛软骨的RNA为模板,按照不同物种BMP4基因的相似性设计特异引物,运用PCR和RACE技术扩增并获得了特异片段,该片段经PCR、酶切和测序验证,证实所克隆序列为牛BMP4的3′端序列,包含有1170bp组成的开放读码框(ORF),编码389个氨基酸,3′非编码区121bp个核苷酸和poly(A)15。同源性分析结果表明,牛BMP4 cDNA最大开放读码框所推测的氨基酸序列与已知人、小鼠、大鼠、狗、羊和鸡等真核生物BMP4氨基酸序列进行比较,分别有94.5%、93.1%、91.9%、87.4%、94.2%、79%的同源性。这为克隆其他物种的BMP4基因提供了依据,同时牛骨形态发生蛋白的测序为我们更好的理解牛的生骨机理提供帮助。 相似文献
104.
Daniela Camargos Costa Ana Paula Madureira Lara Cotta Amaral Bruno Ant?nio Marinho Sanchez Luciano Teixeira Gomes Cor Jésus Fernandes Fontes Jean Ezequiel Limongi Cristiana Ferreira Alves de Brito Luzia Helena Carvalho 《Memórias do Instituto Oswaldo Cruz》2014,109(1):21-28
The polymerase chain reaction (PCR)-based methods for the diagnosis of malaria
infection are expected to accurately identify submicroscopic parasite carriers.
Although a significant number of PCR protocols have been described, few studies have
addressed the performance of PCR amplification in cases of field samples with
submicroscopic malaria infection. Here, the reproducibility of two well-established
PCR protocols (nested-PCR and real-time PCR for the Plasmodium 18
small subunit rRNA gene) were evaluated in a panel of 34 blood field samples from
individuals that are potential reservoirs of malaria infection, but were negative for
malaria by optical microscopy. Regardless of the PCR protocol, a large variation
between the PCR replicates was observed, leading to alternating positive and negative
results in 38% (13 out of 34) of the samples. These findings were quite different
from those obtained from the microscopy-positive patients or the unexposed
individuals; the diagnosis of these individuals could be confirmed based on the high
reproducibility and specificity of the PCR-based protocols. The limitation of PCR
amplification was restricted to the field samples with very low levels of
parasitaemia because titrations of the DNA templates were able to detect < 3
parasites/µL in the blood. In conclusion, conventional PCR protocols require careful
interpretation in cases of submicroscopic malaria infection, as inconsistent and
false-negative results can occur. 相似文献
105.
106.
Matias AC Marinho HS Cyrne L Herrero E Antunes F 《Archives of biochemistry and biophysics》2011,515(1-2):107-111
Taking into account published contradictory results concerning the regulation of fatty acid synthase (Fas) by H(2)O(2), we carried out a systematic study where two methods of H(2)O(2) delivery (steady-state and bolus addition) and the effect of a wide range of H(2)O(2) concentrations were investigated. A decrease in Fas activity was observed for cells exposed to 100 and 150μM H(2)O(2) in a steady-state, while a bolus addition of the same H(2)O(2) concentrations did not alter Fas activity. Similar results were observed for the mRNA levels of FAS1, the gene that encodes Fas subunit β. However, the exposure to a steady-state 50μM H(2)O(2) dose lead to an increase in FAS1 mRNA levels, showing a biphasic modulation of Fas by H(2)O(2). The results obtained emphasize that cellular effects of H(2)O(2) can vary over a narrow range of concentrations. Therefore, a tight control of H(2)O(2) exposure, which can be achieved by exposing H(2)O(2) in a steady-state, is important for cellular studies of H(2)O(2)-dependent redox regulation. 相似文献
107.
Aresta-Branco F Cordeiro AM Marinho HS Cyrne L Antunes F de Almeida RF 《The Journal of biological chemistry》2011,286(7):5043-5054
The plasma membrane of Saccharomyces cerevisiae was studied using the probes trans-parinaric acid and diphenylhexatriene. Diphenylhexatriene anisotropy is a good reporter of global membrane order. The fluorescence lifetimes of trans-parinaric acid are particularly sensitive to the presence and nature of ordered domains, but thus far they have not been measured in yeast cells. A long lifetime typical of the gel phase (>30 ns) was found in wild-type (WT) cells from two different genetic backgrounds, at 24 and 30 °C, providing the first direct evidence for the presence of gel domains in living cells. To understand their nature and location, the study of WT cells was extended to spheroplasts, the isolated plasma membrane, and liposomes from total lipid and plasma membrane lipid extracts (with or without ergosterol extraction by cyclodextrin). It is concluded that the plasma membrane is mostly constituted by ordered domains and that the gel domains found in living cells are predominantly at the plasma membrane and are formed by lipids. To understand their composition, strains with mutations in sphingolipid and ergosterol metabolism and in the glycosylphosphatidylinositol anchor remodeling pathway were also studied. The results strongly indicate that the gel domains are not ergosterol-enriched lipid rafts; they are mainly composed of sphingolipids, possibly inositol phosphorylceramide, and contain glycosylphosphatidylinositol-anchored proteins, suggesting an important role in membrane traffic and signaling, and interactions with the cell wall. The abundance of the sphingolipid-enriched gel domains was inversely related to the cellular membrane system global order, suggesting their involvement in the regulation of membrane properties. 相似文献
108.
MÁRCIA FERNANDES DE AQUINO SANTOS JOSÉ RICARDO M. MERMUDES VERA MARIA MEDINA DA FONSECA 《Palaeontology》2011,54(4):807-814
Abstract: A specimen of Curculionidae (Curculioninae) is described as Arariperhinus monnei gen. et sp. nov. The specimen is preserved on a laminated limestone sample of the Crato Formation (Santana Group), Lower Cretaceous (Aptian–Albian), and was collected from a quarry near Nova Olinda, Chapada do Araripe, State of Ceará, Brazil. The genus is placed in the subfamily Curculioninae because of its strongly convex body and relatively slender rostrum, but mainly by its rounded eyes and lack of a prosternal sulcus and tibial spurs. The very prominent eyes in lateral view, a cylindrical rostrum and a straight posterior margin of ventrite II are strong indications that this fossil belongs to the tribe Anthonomini. However, the claws, which would resolve the exact placement of this fossil, are poorly preserved. Arariperhinus monnei gen. et sp. nov. is distinguishable by the combination of several characters and the first record of the family Curculionidae in the Santana Group; it is the oldest record of a member of the subfamily Curculioninae. 相似文献
109.
110.
RICARDO CRUZ DE CARVALHO ANABELA BERNARDES DA SILVA RENATA SOARES ANDRÉ M. ALMEIDA ANA VARELA COELHO JORGE MARQUES DA SILVA CRISTINA BRANQUINHO 《Plant, cell & environment》2014,37(7):1499-1515
All bryophytes evolved desiccation tolerance (DT) mechanisms during the invasion of terrestrial habitats by early land plants. Are these DT mechanisms still present in bryophytes that colonize aquatic habitats? The aquatic bryophyte Fontinalis antipyretica Hedw. was subjected to two drying regimes and alterations in protein profiles and sucrose accumulation during dehydration and rehydration were investigated. Results show that during fast dehydration, there is very little variation in protein profiles, and upon rehydration proteins are leaked. On the other hand, slow dehydration induces changes in both dehydration and rehydration protein profiles, being similar to the protein profiles displayed by the terrestrial bryophytes Physcomitrella patens (Hedw.) Bruch and Schimp. and, to what is comparable with Syntrichia ruralis (Hedw.) F. Weber and D. Mohr. During dehydration there was a reduction in proteins associated with photosynthesis and the cytoskeleton, and an associated accumulation of proteins involved in sugar metabolism and plant defence mechanisms. Upon rehydration, protein accumulation patterns return to control values for both photosynthesis and cytoskeleton whereas proteins associated with sugar metabolism and defence proteins remain high. The current results suggest that bryophytes from different ecological adaptations may share common DT mechanisms. 相似文献