A polyphasic approach for characterization of a collection of cereal isolates of the Fusarium incarnatum-equiseti species complex

"Available online 22 June 2016"DNA-based phylogenetic analyses have resolved the fungal genus Fusarium into multiple species complexes. The F. incarnatum-equiseti species complex (FIESC) includes fusaria associated with several diseases of agriculturally important crops, including cereals. Although members of FIESC are considered to be only moderately aggressive, they are able to produce a diversity of mycotoxins, including trichothecenes, which can accumulate to harmful levels in cereals. High levels of cryptic speciation have been detected within the FIESC. As a result, it is often necessary to use approaches other than morphological characterization to distinguish species. In the current study, we used a polyphasic approach to characterize a collection of 69 FIESC isolates recovered from cereals in Europe, Turkey, and North America. In a species phylogeny inferred from nucleotide sequences from four housekeeping genes, 65 of the isolates were resolved within the Equiseti clade of the FIESC, and four isolates were resolved within the Incarnatum clade. Seven isolates were resolved as a genealogically exclusive lineage, designated here as FIESC 31. Phylogenies based on nucleotide sequences of trichothecene biosynthetic genes and MALDI-TOF MS (Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry) were largely concordant with phylogeny inferred from the housekeeping gene. Finally, Liquid Chromatography (Time-Of-Flight) Mass Spectrometry [LC-(TOF-)MS(/MS)] revealed variability in mycotoxin production profiles among the different phylogenetic species investigated in this study.This work was supported by the EU project EC KBBE-2007-222690-2 MYCORED

A Strategy for Resolving Europe\u27s Problem Loans

This discussion note uses a new survey of European country authorities and banks to examine the structural obstacles that discourage banks from addressing their problem loans. A three pillared strategy is advocated to remedy the situation, comprising: (i) tightened supervisory policies, (ii) insolvency reforms, and (iii) the development of distressed debt market

Increased vertebral morphometric fracture in patients with postsurgical hypoparathyroidism despite normal bone mineral density

Background\ud In human malaria, the naturally-acquired immune response can result in either the elimination of the parasite or a persistent response mediated by cytokines that leads to immunopathology. The cytokines are responsible for all the symptoms, pathological alterations and the outcome of the infection depends on the reciprocal regulation of the pro and anti-inflammatory cytokines. IL-10 and IFN-gamma are able to mediate this process and their production can be affected by single nucleotide polymorphisms (SNPs) on gene of these cytokines. In this study, the relationship between cytokine IL-10/IFN-gamma levels, parasitaemia, and their gene polymorphisms was examined and the participation of pro-inflammatory and regulatory balance during a natural immune response in Plasmodium vivax-infected individuals was observed.\ud \ud Methods\ud The serum levels of the cytokines IL-4, IL-12, IFN-gamma and IL-10 from 132 patients were evaluated by indirect enzyme-linked immunosorbent assays (ELISA). The polymorphism at position +874 of the IFN-gamma gene was identified by allele-specific polymerase chain reaction (ASO-PCR) method, and the polymorphism at position -1082 of the IL-10 gene was analysed by PCR-RFLP (PCR-Restriction Fragment Length Polymorphism).\ud \ud Results\ud The levels of a pro- (IFN-gamma) and an anti-inflammatory cytokine (IL-10) were significantly higher in P. vivax-infected individuals as compared to healthy controls. The IFN-gamma levels in primoinfected patients were significantly higher than in patients who had suffered only one and more than one previous episode. The mutant alleles of both IFN-gamma and IL-10 genes were more frequent than the wild allele. In the case of the IFNG+874 polymorphism (IFN-gamma) the frequencies of the mutant (A) and wild (T) alleles were 70.13% and 29.87%, respectively. Similar frequencies were recorded in IL-10-1082, with the mutant (A) allele returning a frequency of 70.78%, and the wild (G) allele a frequency of 29.22%. The frequencies of the alleles associated with reduced production of both IFN-gamma and IL-10 were high, but this effect was only observed in the production of IFN-gamma.\ud \ud Conclusions\ud This study has shown evidence of reciprocal regulation of the levels of IL-10 and IFN-gamma cytokines in P. vivax malaria, which is not altered by the presence of polymorphism in the IL-10 gene.M.L.M. received financial support from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and FJAP receive financial support from CNPq (306027/2011-9). This study received financial support from Fundação de Apoio ao Ensino Pesquisa e Assistência (FAEPA). We thank Sebastião Brandão Filho and Marta T. Nakao for technical assistance and Prof. Dr. Edson Z. Martinez for expert contribution on the statistical analysis

Recruitment of MBD1 to target genes requires sequence-specific interaction of the MBD domain with methylated DNA

MBD1, a member of the methyl-CpG-binding domain family of proteins, has been reported to repress transcription of methylated and unmethylated promoters. As some MBD1 isoforms contain two DNA-binding domains—an MBD, which recognizes methylated DNA; and a CXXC3 zinc finger, which binds unmethylated CpG—it is unclear whether these two domains function independently of each other or if they cooperate in facilitating recruitment of MBD1 to particular genomic loci. In this report we investigate DNA-binding specificity of MBD and CXXC3 domains in vitro and in vivo. We find that the methyl-CpG-binding domain of MBD1 binds more efficiently to methylated DNA within a specific sequence context. We identify genes that are targeted by MBD1 in human cells and demonstrate that a functional MBD domain is necessary and sufficient for recruitment of MBD1 to specific sites at these loci, while DNA binding by the CXXC3 motif is largely dispensable. In summary, the binding preferences of MBD1, although dependent upon the presence of methylated DNA, are clearly distinct from those of other methyl-CpG-binding proteins, MBD2 and MeCP2

Evaluation of optimal medical therapy in acute myocardial infarction patients with prior stroke

Background: Treatment of acute myocardial infarction (AMI) patients with prior stroke is a common clinical dilemma. Currently, the application of optimal medical therapy (OMT) and its impact on clinical outcomes are not clear in this patient population. Methods: We retrieved 765 AMI patients with prior stroke who underwent percutaneous coronary intervention (PCI) during the index hospitalization from the international multicenter BleeMACS registry. All of the subjects were divided into two groups based on the prescription they were given prior to discharge. Baseline characteristics and procedural variables were compared between the OMT and non-OMT groups. Mortality, re-AMI, major adverse cardiovascular events (MACE), and bleeding were followed-up for 1 year. Results: Approximately 5% of all patients presenting with AMI were admitted to the hospital for ischemic stroke. Although the prescription rate of each OMT medication was reasonably high (73.3%-97.3%), 47.7% lacked at least one OMT medication. Patients receiving OMT showed a significantly decreased occurrence of mortality (4.5% vs 15.1%, p < 0.001), re-AMI (4.2% vs 9.3%, p = 0.004), and the composite endpoint of death/re-AMI (8.6% vs 20.5%, p < 0.001) compared to those without OMT. No significant difference was observed between the groups regarding bleeding. After adjusting for confounding factors, OMT was the independent protective factor of 1-year mortality, while age was the independent risk factors. Conclusions: OMT at discharge was associated with a significantly lower 1-year mortality of patients with AMI and prior stroke in clinical practice. However, OMT was provided to just half of the eligible patients, leaving room for substantial improvement

Assessment of parental contributions to fast- and slow-growing progenies in the sea bream Sparus aurata L. using a new multiplex PCR

Molecular tools to assist breeding programs in the gilthead sea bream (Sparus aurata L.) are scarce. A new multiplex PCR technique (OVIDORPLEX), which amplifies nine known microsatellite markers, was developed in this work. This multiplex system showed a high mean heterozygosity (>0.800) and a high mean number of alleles per marker (>14) when tested in two sea bream broodstocks (A: 40 breeders and B: 38 breeders). We tested this multiplex PCR for inferring parentage in a Spanish hatchery that graded the animals by size as part of their management procedure. The progeny of the broodstock were divided into fast- and slow-growth groups. Parentage studies revealed that this management procedure entailed a global reduction of the breeders' representation in progeny and that breeders' contributions were significantly unequal. Due to this, effective sample sizes fell to N ê¿13-14 for fast- and N ê¿18-24 for slow-growth progeny groups. These results imply a 3 to 4% rate of inbreeding per generation in the fast-growth group, which is more important to hatchery managers than the slow group. Not all the progeny were evaluated in this experiment (due to the discarding steps), and thus it is difficult to know if the phenotypic performance showed by the fast-growing progeny will be heritable. However, there were genetic differences between the differentiated growth progeny groups (fast vs. slow, F ST values=0.016 to 0.023; P<0.01). We also identified breeders with significantly different contributions to the fast- (10 breeders) or to the slow- (15 breeders) growth progeny groups. Our results demonstrated that this new multiplex PCR could be useful for quantitative programs (breeding programs, detection of QTL, inbreeding control or reconstruction of fish genealogies) to improve the aquaculture of the gilthead sea bream (S. aurata). © 2011 Elsevier B.V.This work was carried out in collaboration with the fish farm Granja Marina Safor, S.L. (Gandia, Valencia, Spain) and the hatchery Piscicultura Marina Mediterranea, S.L. (Burriana, Castellon, Spain). It was financed by JACUMAR (the PROGENSA project) and the Spanish Ministry of Science and Innovation (MICINN; National Program of Resources and Food and Agriculture Technologies, AGL2006-13411-C03-00, SELECTSPARUS, and AGL2007-64040-C03-00, SELECTBREAM, including European Regional Development Funds). V. Gallego was supported by a FPI scholarship financed by MICINN; C. Garcia-Fernandez was supported by a FPU scholarship financed by MICINN; and I. Mazzeo was supported by a FPI scholarship financed by Generalitat Valenciana. We are indebted to three anonymous referees and the journal editor for valuable comments.Borrell, YJ.; Gallego Albiach, V.; García Fernández, C.; Mazzeo ., I.; Pérez Igualada, LM.; Asturiano Nemesio, JF.; Carleos, CE.... (2011). Assessment of parental contributions to fast- and slow-growing progenies in the sea bream Sparus aurata L. using a new multiplex PCR. Aquaculture. 314(1-4):58-65. https://doi.org/10.1016/j.aquaculture.2011.01.028S58653141-

Terfenadine induces apoptosis and autophagy in melanoma cells through ROS-dependent and -independent mechanisms

Previously we found that terfenadine, an H1 histamine receptor antagonist, acts as a potent apoptosis inducer in melanoma cells through modulation of Ca2+ homeostasis. In this report, focusing our attention on the apoptotic mechanisms activated by terfenadine, we show that this drug can potentially activate distinct intrinsic signaling pathways depending on culture conditions. Serum-deprived conditions enhance the cytotoxic effect of terfenadine and caspase-4 and -2 are activated upstream of caspase-9. Moreover, although we found an increase in ROS levels, the apoptosis was ROS independent. Conversely, terfenadine treatment in complete medium induced ROS-dependent apoptosis. Caspase-4, -2, and -9 were simultaneously activated and p73 and Noxa induction were involved. ROS inhibition prevented p73 and Noxa expression but not p53 and p21 expression, suggesting a role for Noxa in p53-independent apoptosis in melanoma cells. Finally, we found that terfenadine induced autophagy, that can promote apoptosis. These findings demonstrate the great potential of terfenadine to kill melanoma cells through different cellular signaling pathways and could contribute to define new therapeutic strategies in melanoma