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891.
Casalino L Magnani D De Falco S Filosa S Minchiotti G Patriarca EJ De Cesare D 《Molecular biotechnology》2012,50(3):171-180
The use of Embryonic Stem Cells (ESCs) holds considerable promise both for drug discovery programs and the treatment of degenerative
disorders in regenerative medicine approaches. Nevertheless, the successful use of ESCs is still limited by the lack of efficient
control of ESC self-renewal and differentiation capabilities. In this context, the possibility to modulate ESC biological
properties and to obtain homogenous populations of correctly specified cells will help developing physiologically relevant
screens, designed for the identification of stem cell modulators. Here, we developed a high throughput screening-suitable
ESC neural differentiation assay by exploiting the Cell
maker robotic platform and demonstrated that neural progenies can be generated from ESCs in complete automation, with high standards
of accuracy and reliability. Moreover, we performed a pilot screening providing proof of concept that this assay allows the
identification of regulators of ESC neural differentiation in full automation. 相似文献
892.
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
893.
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894.
Mendoza M Brito A Schaper DA Spooner VA Alvarado P Castro A Fernández A 《Revista iberoamericana de micología》2012,29(3):120-125
BackgroundSporotrichosis caused by the dimorphic fungus Sporothrix schenckii can presents in a variety of clinical forms. Routine diagnosis is made by mycology and serology studies. Few investigations have been focused on the evaluation of the molecular diagnosis.AimTo determine the value of the nested PCR technique for the diagnosis of experimental sporotrichosis in organs of mice, and to compare the results with the established laboratory diagnostic procedures.MethodsBALB/c mice were inoculated with growing concentrations of the 2 morphological phases of the fungus. The infected animals were sacrificed one month later and specimens from liver, spleen, lung and testicle were obtained to perform wet mount, culture and molecular diagnosis by the nested PCR technique. Blood samples were obtained for determination of specific antibodies against S. schenckii by the double immunodiffusion procedure.ResultsThe pathogenicity observed with the different concentrations of the fungus inoculated and its isolation by culture, showed scarce differences in the study of specimens from organs infected with the 2 morphological phases of S. schenckii. Specimens from organs of mice inoculated with the mycelial phase when studied by wet mount and culture, showed a higher positivity (100 and 37.5%) than those from mice inoculated with the yeast phase (73 and 2%). However, diagnosis by the nested PCR molecular technique applied to the latter specimens showed a higher percentage of positivity (75%) and 43% of positive results coming from animals infected with the mycelial phase. Specific antibody detection was positive in 100% all groups of infected mice.ConclusionsIn the study of experimental sporotrichosis in mice, the culture, as well as the antibody detection, was an effective diagnostic procedure, while the nested PCR and microscopic studies had a lower diagnostic value. 相似文献
895.
Rodolfo Valdés Andrés Tamayo Marcos González Sigifredo Padilla Déborah Geada William Ferro Lorely Milá Leonardo Gómez Rosario Alemán Alberto Leyva Cristina García Otto Mendoza Tatiana Alvarez Lamay Dorta Yanet Villega Dobián Cecilia Hasel Aragón Tatiana González Maylín La O Jorge López 《Biotechnology and Bioprocess Engineering》2012,17(1):145-159
Monoclonal antibody (mAb) production methods (ascites, in vitro technologies, transgenic animals, and dicot or monocot transgenic plants; moss, algae) have been improved since they were first developed in 1975. In this study, we illustrate a summary of a study case in which mice, a hollow fiber system, and tobacco transgenic plants were assessed for the production of mAb for vaccine manufacturing and vaccine production. 相似文献
896.
Borja Jiménez-Alfaro Eduardo Fernández-Pascual Tomás E. Díaz González Aaron Pérez-Haase Josep M. Ninot 《Folia Geobotanica》2012,47(4):403-419
In temperate mountains, fens have been reported as relict habitats subject to geographical fragmentation and broad climatic gradients, but few studies have analyzed the influence of these factors on plant diversity. Here we investigate the effect of isolation on the vegetation diversity of rich fens (Caricion davallianae) in the mountains of the Iberian Peninsula, the distribution limit of these habitats in south-western Europe. We used plot-based vegetation data from the Pyrenees and the Cantabrian mountain range to evaluate their regional species-pool, occurrence of specialists, beta-diversity and the effect of geo-climatic variables on their species-richness and species-composition. We found a lower ratio of rare specialists in the Pyrenees than in the Cantabrian range, but similar estimates in the species pools, total species-richness per plot and beta-diversity. The isolation of the two mountain regions resulted in different species assemblages best predicted by summer precipitation and bedrock types, showing region-based differences in the response of vegetation and plant specialists to the environment. The tighter correlation between local climate and diversity estimates in the Cantabrian range suggests relict character of rich fens in that region, where climatic conditions have restricted local distribution of formerly more widely distributed specialists. Although there is no relevant evidence of vegetation impoverishment in that region, historical isolation has probably resulted in the existence of fragmentary plant communities. We conclude that fen vegetation may experience long-time persistence in climatically sub-optimal mountain refugia, but related plant specialists may be sensitive to climatic changes and subject to the extinction of local populations. 相似文献
897.
Ariadne Letra Walid Fakhouri Renata F. Fonseca Renato Menezes Inga Kempa Joanne L. Prasad Toby G. McHenry Andrew C. Lidral Lina Moreno Jeffrey C. Murray Sandra Daack-Hirsch Mary L. Marazita Eduardo E. Castilla Baiba Lace Ieda M. Orioli Jose M. Granjeiro Brian C. Schutte Alexandre R. Vieira 《PloS one》2012,7(9)
Previous evidence from tooth agenesis studies suggested IRF6 and TGFA interact. Since tooth agenesis is commonly found in individuals with cleft lip/palate (CL/P), we used four large cohorts to evaluate if IRF6 and TGFA interaction contributes to CL/P. Markers within and flanking IRF6 and TGFA genes were tested using Taqman or SYBR green chemistries for case-control analyses in 1,000 Brazilian individuals. We looked for evidence of gene-gene interaction between IRF6 and TGFA by testing if markers associated with CL/P were overtransmitted together in the case-control Brazilian dataset and in the additional family datasets. Genotypes for an additional 142 case-parent trios from South America drawn from the Latin American Collaborative Study of Congenital Malformations (ECLAMC), 154 cases from Latvia, and 8,717 individuals from several cohorts were available for replication of tests for interaction. Tgfa and Irf6 expression at critical stages during palatogenesis was analyzed in wild type and Irf6 knockout mice. Markers in and near IRF6 and TGFA were associated with CL/P in the Brazilian cohort (p<10−6). IRF6 was also associated with cleft palate (CP) with impaction of permanent teeth (p<10−6). Statistical evidence of interaction between IRF6 and TGFA was found in all data sets (p = 0.013 for Brazilians; p = 0.046 for ECLAMC; p = 10−6 for Latvians, and p = 0.003 for the 8,717 individuals). Tgfa was not expressed in the palatal tissues of Irf6 knockout mice. IRF6 and TGFA contribute to subsets of CL/P with specific dental anomalies. Moreover, this potential IRF6-TGFA interaction may account for as much as 1% to 10% of CL/P cases. The Irf6-knockout model further supports the evidence of IRF6-TGFA interaction found in humans. 相似文献
898.
Eduardo Díaz-Rubio Auxiliadora Gómez-Espa?a Bartomeu Massutí Javier Sastre Margarita Reboredo José Luis Manzano Fernando Rivera MaJosé Safont Clara Montagut Encarnación González Manuel Benavides Eugenio Marcuello Andrés Cervantes Purificación Martínez de Prado Carlos Fernández-Martos Antonio Arrivi Inmaculada Bando Enrique Aranda Spanish Cooperative Group for the Treatment of Digestive Tumors 《PloS one》2012,7(10)
Background
In the MACRO study, patients with metastatic colorectal cancer (mCRC) were randomised to first-line treatment with 6 cycles of capecitabine and oxaliplatin (XELOX) plus bevacizumab followed by either single-agent bevacizumab or XELOX plus bevacizumab until disease progression. An additional retrospective analysis was performed to define the prognostic value of tumour KRAS status on progression-free survival (PFS), overall survival (OS) and response rates.Methodology/Principal Findings
KRAS data (tumour KRAS status and type of mutation) were collected by questionnaire from participating centres that performed KRAS analyses. These data were then cross-referenced with efficacy data for relevant patients in the MACRO study database. KRAS status was analysed in 394 of the 480 patients (82.1%) in the MACRO study. Wild-type (WT) KRAS tumours were found in 219 patients (56%) and mutant (MT) KRAS in 175 patients (44%). Median PFS was 10.9 months for patients with WT KRAS and 9.4 months for patients with MT KRAS tumours (p = 0.0038; HR: 1.40; 95% CI:1.12–1.77). The difference in OS was also significant: 26.7 months versus 18.0 months for WT versus MT KRAS, respectively (p = 0.0002; HR: 1.55; 95% CI: 1.23–1.96). Univariate and multivariate analyses showed that KRAS was an independent variable for both PFS and OS. Responses were observed in 126 patients (57.5%) with WT KRAS tumours and 76 patients (43.4%) with MT KRAS tumours (p = 0.0054; OR: 1.77; 95% CI: 1.18–2.64).Conclusions/Significance
This analysis of the MACRO study suggests a prognostic role for tumour KRAS status in patients with mCRC treated with XELOX plus bevacizumab. For both PFS and OS, KRAS status was an independent factor in univariate and multivariate analyses. 相似文献899.
Takehiko Shimizu Bao Ho Kathleen Deeley Jessica Brise?o-Ruiz Italo M. Faraco Jr Brett I. Schupack Jo?o A. Brancher Giovana D. Pecharki Erika C. Küchler Patricia N. Tannure Andrea Lips Thays C. S. Vieira Asli Patir Mine Yildirim Fernando A. Poletta Juan C. Mereb Judith M. Resick Carla A. Brandon Iêda M. Orioli Eduardo E. Castilla Mary L. Marazita Figen Seymen Marcelo C. Costa José M. Granjeiro Paula C. Trevilatto Alexandre R. Vieira 《PloS one》2012,7(9)
There is evidence for a genetic component in caries susceptibility, and studies in humans have suggested that variation in enamel formation genes may contribute to caries. For the present study, we used DNA samples collected from 1,831 individuals from various population data sets. Single nucleotide polymorphism markers were genotyped in selected genes (ameloblastin, amelogenin, enamelin, tuftelin, and tuftelin interacting protein 11) that influence enamel formation. Allele and genotype frequencies were compared between groups with distinct caries experience. Associations with caries experience can be detected but they are not necessarily replicated in all population groups and the most expressive results was for a marker in AMELX (p = 0.0007). To help interpret these results, we evaluated if enamel microhardness changes under simulated cariogenic challenges are associated with genetic variations in these same genes. After creating an artificial caries lesion, associations could be seen between genetic variation in TUFT1 (p = 0.006) and TUIP11 (p = 0.0006) with enamel microhardness. Our results suggest that the influence of genetic variation of enamel formation genes may influence the dynamic interactions between the enamel surface and the oral cavity. 相似文献
900.
Benita Wolf Adrian Schwarzer Anik L. C?té Thomas H. Hampton Thomas Schwaab Eduardo Huarte Craig R. Tomlinson Jiang Gui Jan L. Fisher Camilo E. Fadul Joshua W. Hamilton Marc S. Ernstoff 《PloS one》2012,7(12)
Lymphocytes are a key component of the immune system and their differentiation and function are directly influenced by cancer. We examined peripheral blood lymphocyte (PBL) gene expression as a biomarker of illness and treatment effect using the Affymetrix Human Gene ST1 platform in patients with metastatic renal cell carcinoma (mRCC) who received combined treatment with IL-2, interferon-?-2a and dendritic cell vaccine. We examined gene expression, cytokine levels in patient serum and lymphocyte subsets as determined by flow cytometry (FCM). Pre-treatment PBLs from patients with mRCC exhibit a gene expression profile and serum cytokine profile consistent with inflammation and proliferation not found in healthy donors (HD). PBL gene expression from patients with mRCC showed increased mRNA of genes involved with T-cell and TREG-cell activation pathways, which was also reflected in lymphocyte subset distribution. Overall, PBL gene expression post-treatment (POST) was not significantly different than pre-treatment (PRE). Nevertheless, treatment related changes in gene expression (post-treatment minus pre-treatment) revealed an increased expression of T-cell and B-cell receptor signaling pathways in responding (R) patients compared to non-responding (NR) patients. In addition, we observed down-regulation of TREG-cell pathways post-treatment in R vs. NR patients. While exploratory in nature, this study supports the hypothesis that enhanced inflammatory cytotoxic pathways coupled with blunting of the regulatory pathways is necessary for effective anti-cancer activity associated with immune therapy. This type of analysis can potentially identify additional immune therapeutic targets in patients with mRCC. 相似文献