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141.
Pereira F Carneiro J Matthiesen R van Asch B Pinto N Gusmão L Amorim A 《Nucleic acids research》2010,38(22):e203
The quest for a universal and efficient method of identifying species has been a longstanding challenge in biology. Here, we show that accurate identification of species in all domains of life can be accomplished by multiplex analysis of variable-length sequences containing multiple insertion/deletion variants. The new method, called SPInDel, is able to discriminate 93.3% of eukaryotic species from 18 taxonomic groups. We also demonstrate that the identification of prokaryotic and viral species with numeric profiles of fragment lengths is generally straightforward. A computational platform is presented to facilitate the planning of projects and includes a large data set with nearly 1800 numeric profiles for species in all domains of life (1556 for eukaryotes, 105 for prokaryotes and 130 for viruses). Finally, a SPInDel profiling kit for discrimination of 10 mammalian species was successfully validated on highly processed food products with species mixtures and proved to be easily adaptable to multiple screening procedures routinely used in molecular biology laboratories. These results suggest that SPInDel is a reliable and cost-effective method for broad-spectrum species identification that is appropriate for use in suboptimal samples and is amenable to different high-throughput genotyping platforms without the need for DNA sequencing. 相似文献
142.
Fgf and Tgfβ are key regulators of bone development. It is not known, however, whether there is a relationship between defective Fgf signalling, resulting in a premature cranial suture fusion, and Tgfβ signalling. We used mouse calvaria osteoblasts carrying a mutation (hFGFR2-C278F) associated with Crouzon and Pfeiffer syndromes to investigate effects of this mutation on cell growth and possible mechanisms underlying it. Mutated osteoblasts displayed reduced S-phase, increased apoptosis and increased differentiation. As Tgfβ signalling appeared to be required in an autocrine/paracrine manner for osteoblast proliferation, we tested the hypothesis that reduced growth might be due, at least in part, to an altered balance between FGF and Tgfβ signalling. Tgfβ expression was indeed decreased in mutated osteoblasts, as compared to osteoblasts carrying the wild type hFGFR2. Treatment with Tgfβ, however, neither increased proliferation in mutated osteoblasts, unlike in controls, nor rescued proliferation in control osteoblasts treated with an Erk1/2 inhibitor. Significantly, Erk2, that is important for proliferation, was reduced relatively to Erk1 in mutated cells. Altogether this study suggests that the hFGFR2-C278F mutation affects the osteoblast ability to respond to Tgfβ stimulation via the Erk pathway and that the overall effect of the mutation is a loss of function. 相似文献
143.
J. E. Martínez J. F. Calvo J. A. Martínez I. Zuberogoitia E. Cerezo J. Manrique G. J. Gómez J. C. Nevado M. Sánchez R. Sánchez J. Bayo A. Pallarés C. González J. M. Gómez P. Pérez J. Motos 《Biodiversity and Conservation》2010,19(13):3757-3767
Although wind farms in Spain have increased in numbers in recent years, their impact on birds, particularly large raptors, has received relatively little attention in the scientific literature. We study the potential impact of 72 wind energy developments planned for the south-east of Spain covering 128 golden eagle and 152 Bonelli’s eagle territories using nearest neighbour distances (NND) as an indicator of potential future interactions (abandonment, displacement and collision risk). Our analyses indicate low levels of potential interactions between wind farms and large eagles, and suggest that, of the two species studied, golden eagles will be the more affected because a greater proportion of wind farms will be constructed close to the breeding territories of this species. In the light of these findings, we discuss various management strategies in order to improve the compatibility of harvesting wind energy with the conservation of both species. 相似文献
144.
Xuemei Zhao Katie Southwick Helene L. Cardasis Yi Du Michael E. Lassman Dan Xie Mohamed El‐Sherbeini Wayne M. Geissler KellyAnn D. Pryor Andreas Verras Margarita Garcia‐Calvo Dong‐Ming Shen Nathan A. Yates Shirly Pinto Ronald C. Hendrickon 《Proteomics》2010,10(15):2882-2886
Prolylcarboxypeptidase (PRCP) is a serine protease that catalyzes the cleavage of C‐terminal amino acids linked to proline in peptides. It is ubiquitously expressed and is involved in regulating blood pressure, proliferation, inflammation, angiogenesis, and weight maintenance. To identify the candidate proximal target engagement markers for PRCP inhibition in the central nervous system, we profiled the peptidome of human cerebrospinal fluid to look for PRCP substrates using a MS‐based in vitro substrate profiling assay. These experiments identified a single peptide, with the sequence YPRPIHPA, as a novel substrate for PRCP in human cerebrospinal fluid. The peptide YPRPIHPA is from the extracellular portion of human endothelin B receptor‐like protein 2. 相似文献
145.
Estefania Peña B. Calvo M. A. Martínez P. Martins T. Mascarenhas R. M. N. Jorge A. Ferreira M. Doblaré 《Biomechanics and modeling in mechanobiology》2010,9(1):35-44
In this paper, the viscoelastic mechanical properties of vaginal tissue are investigated. Using previous results of the authors
on the mechanical properties of biological soft tissues and newly experimental data from uniaxial tension tests, a new model
for the viscoelastic mechanical properties of the human vaginal tissue is proposed. The structural model seems to be sufficiently
accurate to guarantee its application to prediction of reliable stress distributions, and is suitable for finite element computations.
The obtained results may be helpful in the design of surgical procedures with autologous tissue or prostheses. 相似文献
146.
Monsalve-Castillo F Chacín-Bonilla L Atencio RJ Espinoza LP Costa-León L Echevarría JM 《Memórias do Instituto Oswaldo Cruz》2007,102(1):107-110
Previous studies have not found hepatitis C virus (HCV) infection in Amerindians from Western Venezuela. A survey of 254 Bari and Yukpa natives aged 10-60 years (mean +/- SD age = 35 +/- 5.4 years) from four communities, two Bari and two Yukpa, in this area were studied to assess the prevalence of antibodies to HCV (anti-HCV) and HCV RNA among these indigenous populations. Serum samples were examined initially for anti-HCV by a four generation enzyme-linked immunosorbent assay (ELISA). Reactive samples were then tested using a third generation recombinant immunoblot assay (RIBA-3). Viral RNA was investigated in all immunoblot-reactive samples by a nested polymerase chain reaction (PCR) method. Six (2.3%) of 254 natives were positive by ELISA, one (2.2%) of these reactive samples were positive by RIBA, and four (1.5%) were indeterminate. Only two (0.8%) were positive by PCR, corresponding to 1 (2.1%) of 47 inhabitants of a Yukpa community and to 1 (2.2%) of 45 subjects of a Bari community. Iatrogenic is thought to play a role in acquisition of the infection. The findings indicate a HCV focus of low endemicity and are compatible with a low degree of exposures of the natives to the virus. Studies are necessary to assess the risk factors for infection in these Amerindians. 相似文献
147.
Inflammatory bowel disease (IBD) is a chronic intestinal inflammatory disorder considered as a consequence of an aberrant response of the immune system to luminal antigens. Numerous groups of agents are being evaluated as novel therapeutic approaches for its treatment; in this way, different peptides have emerged as potential candidates. Galanin is an active neuropeptide distributed in the central and periphery nervous systems although it has been also described having important autocrine and paracrine regulatory capacities with interesting inflammatory and immune properties. In this line, we have observed that galanin treatment has a significant preventive effect in the experimental trinitrobenzensulfonic acid (TNBS) acute model of inflammatory colitis. The aim of the present study was to investigate intensively the role played by the peptide in the evolution of the inflammatory pathology associated to IBD. Galanin (5 and 10 microg/kg/day) was administered i.p., daily, starting 24 h after TNBS instillation, and continuing for 14 and 21 days. The lesions were blindly scored according to macroscopic and histological analyses and quantified as ulcer index. The results demonstrated that chronic administration of galanin improved the colon injury than the TNBS induced. The study by Western-blotting of the expression of nitric oxide inducible enzyme (iNOS), as well as the total nitrite production (NO) assayed by Griess-reaction, showed significant reduction associated with peptide administration. The number of mast cells was also identified in histological preparations stained with toluidine blue and the results showed that samples from galanin treatment, mostly at 21 days, had increased the number of these cells and many of them had a degranulated feature. In conclusion, chronic administration of galanin is able to exert a beneficial effect in the animal model of IBD assayed improving the reparative process. Participation of nitric oxide pathways and mucosal mast cells can not be discarded. 相似文献
148.
Cary JW OBrian GR Nielsen DM Nierman W Harris-Coward P Yu J Bhatnagar D Cleveland TE Payne GA Calvo AM 《Applied microbiology and biotechnology》2007,76(5):1107-1118
The aflatoxin-producing fungi, Aspergillus flavus and A. parasiticus, form structures called sclerotia that allow for survival under adverse conditions. Deletion of the veA gene in A. flavus and A. parasiticus blocks production of aflatoxin as well as sclerotial formation. We used microarray technology to identify genes differentially
expressed in wild-type veA and veA mutant strains that could be involved in aflatoxin production and sclerotial development in A. flavus. The DNA microarray analysis revealed 684 genes whose expression changed significantly over time; 136 of these were differentially
expressed between the two strains including 27 genes that demonstrated a significant difference in expression both between
strains and over time. A group of 115 genes showed greater expression in the wild-type than in the veA mutant strain. We identified a subgroup of veA-dependent genes that exhibited time-dependent expression profiles similar to those of known aflatoxin biosynthetic genes
or that were candidates for involvement in sclerotial production in the wild type. 相似文献
149.
Calcium channels must be subjected to a very precise regulation in order to preserve cell function and viability. Voltage gated calcium channels (VGCC) represent the main pathway for calcium entry in excitable cells. This explains why depolarization induces a rapid-onset and short-term inactivation of calcium currents. Contrarily to this well-documented mechanism to maintain calcium below toxic levels, the regulatory pathways inducing longer-lasting changes and cell surface expression of functional calcium channels are largely unknown. Since calcium is a main player in the activity-dependent regulation of many genes, we hypothesize that calcium channel coding genes could be also subjected to activity-dependent regulation. We have used prolonged depolarization to analyze the effects of sustained intracellular calcium elevation on the mRNAs coding for the different alpha(1) pore-forming subunits of the calcium channels expressed in chromaffin cells. Our findings reveal that persistent depolarization is accompanied by a prolonged intracellular calcium elevation and reduction of calcium current. This calcium current inhibition could be mediated, at least partially, by the downregulation of the mRNAs coding for several alpha(1) subunits. Thus, we show here that depolarization inhibits the expression of Ca(V)1.1, Ca(V)1.2, Ca(V)1.3, Ca(V)2.2 and Ca(V)2.3 mRNAs, while the Ca(V)2.1 mRNA remains unmodified. Moreover, such downregulation of channels depends on calcium entry through the L-type calcium channel, as both mRNA and calcium current changes induced by depolarization are abrogated by L-type channel specific blockers. 相似文献
150.
Fernández-Acero FJ Jorge I Calvo E Vallejo I Carbú M Camafeita E Garrido C López JA Jorrin J Cantoral JM 《Archives of microbiology》2007,187(3):207-215
Botrytis cinerea is a phytopathogenic fungus causing disease in a substantial number of economically important crops. In an attempt to identify putative fungal virulence factors, the two-dimensional gel electrophoresis (2-DE) protein profile
from two B. cinerea strains differing in virulence and toxin production were compared. Protein extracts from fungal mycelium obtained by tissue
homogenization were analyzed. The mycelial 2-DE protein profile revealed the existence of qualitative and quantitative differences
between the analyzed strains. The lack of genomic data from B. cinerea required the use of peptide fragmentation data from MALDI-TOF/TOF and ESI ion trap for protein identification, resulting
in the identification of 27 protein spots. A significant number of spots were identified as malate dehydrogenase (MDH) and
glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The different expression patterns revealed by some of the identified proteins
could be ascribed to differences in virulence between strains. Our results indicate that proteomic analysis are becoming an
important tool to be used as a starting point for identifying new pathogenicity factors, therapeutic targets and for basic
research on this plant pathogen in the postgenomic era. 相似文献