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11.
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
12.
Progeny From Irradiated Colorectal Cancer Cells Acquire an EMT‐Like Phenotype and Activate Wnt/β‐Catenin Pathway
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Lilian Gonçalves dos Reis Bastos Priscila Guimarães de Marcondes Julio Cesar Madureira de‐Freitas‐Junior Fernanda Leve André Luiz Mencalha Waldemir Fernandes de Souza Wallace Martins de Araujo Marcelo Neves Tanaka Eliana Saul Furquim Werneck Abdelhay José Andrés Morgado‐Díaz 《Journal of cellular biochemistry》2014,115(12):2175-2187
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Yanwen Jiang David Redmond Kui Nie Ken W Eng Thomas Clozel Peter Martin Leonard HC Tan Ari M Melnick Wayne Tam Olivier Elemento 《Genome biology》2014,15(8)
Background
Molecular mechanisms associated with frequent relapse of diffuse large B-cell lymphoma (DLBCL) are poorly defined. It is especially unclear how primary tumor clonal heterogeneity contributes to relapse. Here, we explore unique features of B-cell lymphomas - VDJ recombination and somatic hypermutation - to address this question.Results
We performed high-throughput sequencing of rearranged VDJ junctions in 14 pairs of matched diagnosis-relapse tumors, among which 7 pairs were further characterized by exome sequencing. We identify two distinctive modes of clonal evolution of DLBCL relapse: an early-divergent mode in which clonally related diagnosis and relapse tumors diverged early and developed in parallel; and a late-divergent mode in which relapse tumors developed directly from diagnosis tumors with minor divergence. By examining mutation patterns in the context of phylogenetic information provided by VDJ junctions, we identified mutations in epigenetic modifiers such as KMT2D as potential early driving events in lymphomagenesis and immune escape alterations as relapse-associated events.Conclusions
Altogether, our study for the first time provides important evidence that DLBCL relapse may result from multiple, distinct tumor evolutionary mechanisms, providing rationale for therapies for each mechanism. Moreover, this study highlights the urgent need to understand the driving roles of epigenetic modifier mutations in lymphomagenesis, and immune surveillance factor genetic lesions in relapse.Electronic supplementary material
The online version of this article (doi:10.1186/s13059-014-0432-0) contains supplementary material, which is available to authorized users. 相似文献15.
Aline Lima Leite Janete Gualiume Vaz Madureira Lobo Heloísa Aparecida Barbosa da Silva Pereira Mileni Silva Fernandes Tatiani Martini Fernanda Zucki Dóris Hissako Sumida Alfredo Rigalli Marília Afonso Rabelo Buzalaf 《PloS one》2014,9(9)
Administration of high doses of fluoride (F) can alter glucose homeostasis and lead to insulin resistance (IR). This study determined the profile of protein expression in the gastrocnemius muscle of rats with streptozotocin-induced diabetes that were chronically exposed to F. Male Wistar rats (60 days old) were randomly distributed into two groups of 18 animals. In one group, diabetes was induced through the administration of streptozotocin. Each group (D-diabetic and ND-non-diabetic) was further divided into 3 subgroups each of which was exposed to a different F concentration via drinking water (0 ppm, 10 ppm or 50 ppm F, as NaF). After 22 days of treatment, the gastrocnemius muscle was collected and submitted to proteomic analysis (2D-PAGE followed by LC-MS/MS). Protein functions were classified by the GO biological process (ClueGO v2.0.7+Clupedia v1.0.8) and protein-protein interaction networks were constructed (PSICQUIC, Cytoscape). Quantitative intensity analysis of the proteomic data revealed differential expression of 75 spots for ND0 vs. D0, 76 for ND10 vs.D10, 58 spots for ND50 vs. D50, 52 spots for D0 vs. D10 and 38 spots for D0 vs. D50. The GO annotations with the most significant terms in the comparisons of ND0 vs. D0, ND10 vs. D10, ND50 vs. D50, D0 vs. D10 and D0 vs. D50, were muscle contraction, carbohydrate catabolic processes, generation of precursor metabolites and energy, NAD metabolic processes and gluconeogenesis, respectively. Analysis of subnetworks revealed that, in all comparisons, proteins with fold changes interacted with GLUT4. GLUT4 interacting proteins, such as MDH and the stress proteins HSPB8 and GRP78, exhibited decreased expression when D animals were exposed to F. The presence of the two stress proteins indicates an increase in IR, which might worsen diabetes. Future studies should evaluate whether diabetic animals treated with F have increased IR, as well as which molecular mechanisms are involved. 相似文献
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Waldemir Fernandes de Souza Leandro Augusto Barbosa Lijun Liu Wallace Martins de Araujo Julio Cesar Madureira de-Freitas-Junior Natalia Fortunato-Miranda Carlos Frederico Leite Fontes José Andrés Morgado-Díaz 《The Journal of membrane biology》2014,247(1):23-33
Studies have reported that Na,K-ATPase interacts with E-cadherin to stabilize (AJs) and regulate the expression of claudins, the main proteins present in the tight junction (TJ) in epithelial cells containing caveolae. However, the role of this ATPase in the regulation of the AJ and TJ proteins in colorectal cancer cells as well as the molecular events underlying this event in a caveolae-independent system remain undefined. In the present study, we used ouabain, a classic drug known to inhibit Na,K-ATPase, and Caco-2 cells, which are a well-established human colorectal cancer model that does not exhibit caveolae. We demonstrated that ouabain treatment resulted in a reduction of the β1 Na,K-ATPase protein and cell redistribution of the AJ proteins E-cadherin and β-catenin, as well as the α1 Na,K-ATPase subunit. Furthermore, ouabain increased claudin-3 protein levels, impaired the TJ barrier function and increased cell viability and proliferation during the early stages of treatment. Additionally, the observed ouabain-induced events were dependent on the activation of ERK1/2 signaling; but in contrast to previous studies, this signaling cascade was caveolae-independent. In conclusion, our findings strongly suggest that α1 and β1 Na,K-ATPase downregulation and ERK1/2 activation induced by ouabain are interlinked events that play an important role during cell–cell adhesion loss, which is an important step during the tumor progression of colorectal carcinomas. 相似文献
17.
In order to study how N, P, Fe, Mn and S concentrations in pore waters change with time at different temperatures, an incubation experiment was carried out with surficial intertidal sediment. To evaluate the importance of benthic microorganisms, an abiotic control was established by poisoning sediment. The live and poisoned sediments were incubated for ten hours at 10, 21, 30 and 40°C. Dissolved Inorganic Nitrogen (DIN), Dissolved Reactive Phosphorus (DRP), NH4
+, total dissolved manganese (Mndiss), total dissolved iron (Fediss) and soluble inorganic sulphide (HS–)t were followed in the pore water samples. Results indicated that high temperature influenced nitrification, allowing accumulation of ammonia and that microorganism activity did not seem important for Mn reduction. Anaerobic nitrification by Mn reduction was advanced as an explanation of the behaviour of DIN during the experiment. 相似文献
18.
Vivian Oliveira Moura-Massari Flávia Siqueira Cunha Larissa Garcia Gomes Diogo Bugano Diniz Gomes José Ant?nio Miguel Marcondes Guiomar Madureira Berenice Bilharinho de Mendonca Tania A. Sartori Sanchez Bachega 《PloS one》2016,11(2)
Background
In the nonclassical form (NC), good correlation has been observed between genotypes and 17OH-progesterone (17-OHP) levels. However, this correlation was not identified with regard to the severity of hyperandrogenic manifestations, which could depend on interindividual variability in peripheral androgen sensitivity. Androgen action is modulated by the polymorphic CAG tract (nCAG) of the androgen receptor (AR) gene and by polymorphisms in 5α-reductase type 2 (SRD5A2) enzyme, both of which are involved in the severity of hyperandrogenic disorders.Objectives
To analyze whether nCAG-AR and SRD5A2 polymorphisms influence the severity of the nonclassical phenotype.Patients
NC patients (n = 114) diagnosed by stimulated-17OHP ≥10 ng/mL were divided into groups according to the beginning of hyperandrogenic manifestations (pediatric and adolescent/adult) and CYP21A2 genotypes (C/C: homozygosis for mild mutations; A/C: compound heterozygosis for severe/mild mutations).Methods
CYP21A2 mutations were screened by allelic-specific PCR, MLPA and/or sequencing. HpaII-digested and HpaII-undigested DNA samples underwent GeneScan analysis to study nCAG, and the SRD5A2 polymorphisms were screened by RLFP.Results
Mean nCAG did not differ among pediatric, adolescent/adult and asymptomatic subjects. In the C/C genotype, we observed a significantly lower frequency of longer CAG alleles in pediatric patients than in adolescent/adults (p = 0.01). In patients carrying the A/C genotype, the frequencies of shorter and longer CAG alleles did not differ between pediatric patients and adolescent/adults (p>0.05). Patients with clitoromegaly had significantly lower weighted CAG biallelic mean than those without it: 19.1±2.7 and 21.6±2.5, respectively (p = 0.007), independent of the CYP21A2 genotype''s severity. The SRD5A2 polymorphisms were not associated with the variability of hyperandrogenic NC phenotypes.Conclusions
In this series, we observed a modulatory effect of the CAG-AR tract on clinical manifestations of the NC form. Although the NC form is a monogenic disorder, our preliminary data suggested that the interindividual variability of the hyperandrogenic phenotype could arise from polygenic interactions. 相似文献19.
J Brand?o Néto B B de Mendon?a T Shuhama J S Marchini G Madureira W P Pimenta M T Tornero 《Hormones et métabolisme》1989,21(4):203-206
The response of plasma prolactin (PRL) to oral administration of increasing doses of zinc (25.0, 37.5 and 50.0 mg) was studied in 17 normal adult men and women. Blood samples were collected at 10 and 30-min intervals over a period of 120 min after two basal times (-30 and 0 min). PRL concentrations significantly fell below basal levels in all subjects in response to the increase in plasma zinc levels, as compared to the controls. These results suggest that acute hyperzincemia can inhibit basal PRL secretion in normal individuals. 相似文献
20.