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1.
Circ J ; 85(9): 1494-1504, 2021 08 25.
Artigo em Inglês | MEDLINE | ID: mdl-33980765

RESUMO

BACKGROUND: Wild-type transthyretin-related amyloidosis (ATTRwt) and degenerative aortic stenosis (AS) are both age-related. Diagnosis of cardiac amyloidosis (CA) among patients with AS may be difficult due to overlapping morphological and functional criteria. The aim of this study was to describe an echocardiographic longitudinal strain (LS) pattern among patients with AS with and without ATTRwt.Methods and Results:Patients who have AS with ATTRwt (n=30), AS without ATTRwt (n=50) and ATTRwt without AS (n=31) underwent two-dimensional speckle-tracking echocardiography. Transthyretin CA was based on positive bone scintigraphy without monoclonal gammopathy. All patients showed a gradual decrease in LS from the base to the apex resulting in a decrease of the global LS. A cut-off value of 1.0 for relative apical LS (average apical LS/[average basal LS+mid-LS]) was sensitive (88%) but less specific (68%) in differentiating ATTRwt among patients with severe AS. The best cut-off value for relative apical LS for identifying patients with ATTRwt among the whole population was 0.9 (sensitivity 74%, specificity 66%); however, 35%, 25% and 11% of patients who have ATTRwt without AS, with moderate AS and with severe AS, respectively, did not reach this threshold. CONCLUSIONS: A decrease of global and relative apical LS is common in patients with AS, even in the absence of ATTRwt. ATTRwt CA can be present even in the absence of relative apical sparing of LS.


Assuntos
Neuropatias Amiloides Familiares , Estenose da Valva Aórtica , Neuropatias Amiloides Familiares/complicações , Neuropatias Amiloides Familiares/diagnóstico por imagem , Estenose da Valva Aórtica/diagnóstico por imagem , Ecocardiografia/métodos , Humanos , Tomografia Computadorizada por Raios X
2.
BMC Genomics ; 18(1): 667, 2017 Aug 29.
Artigo em Inglês | MEDLINE | ID: mdl-28851275

RESUMO

BACKGROUND: The ascomycete fungus Colletotrichum higginsianum causes anthracnose disease of brassica crops and the model plant Arabidopsis thaliana. Previous versions of the genome sequence were highly fragmented, causing errors in the prediction of protein-coding genes and preventing the analysis of repetitive sequences and genome architecture. RESULTS: Here, we re-sequenced the genome using single-molecule real-time (SMRT) sequencing technology and, in combination with optical map data, this provided a gapless assembly of all twelve chromosomes except for the ribosomal DNA repeat cluster on chromosome 7. The more accurate gene annotation made possible by this new assembly revealed a large repertoire of secondary metabolism (SM) key genes (89) and putative biosynthetic pathways (77 SM gene clusters). The two mini-chromosomes differed from the ten core chromosomes in being repeat- and AT-rich and gene-poor but were significantly enriched with genes encoding putative secreted effector proteins. Transposable elements (TEs) were found to occupy 7% of the genome by length. Certain TE families showed a statistically significant association with effector genes and SM cluster genes and were transcriptionally active at particular stages of fungal development. All 24 subtelomeres were found to contain one of three highly-conserved repeat elements which, by providing sites for homologous recombination, were probably instrumental in four segmental duplications. CONCLUSION: The gapless genome of C. higginsianum provides access to repeat-rich regions that were previously poorly assembled, notably the mini-chromosomes and subtelomeres, and allowed prediction of the complete SM gene repertoire. It also provides insights into the potential role of TEs in gene and genome evolution and host adaptation in this asexual pathogen.


Assuntos
Cromossomos Fúngicos/genética , Colletotrichum/genética , Colletotrichum/metabolismo , Elementos de DNA Transponíveis/genética , Genômica , Família Multigênica/genética , Recombinação Homóloga/genética , Anotação de Sequência Molecular , Filogenia , Mutação Puntual/genética
3.
New Phytol ; 211(4): 1323-37, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27174033

RESUMO

The genome of the hemibiotrophic anthracnose fungus, Colletotrichum higginsianum, encodes a large repertoire of candidate-secreted effectors containing LysM domains, but the role of such proteins in the pathogenicity of any Colletotrichum species is unknown. Here, we characterized the function of two effectors, ChELP1 and ChELP2, which are transcriptionally activated during the initial intracellular biotrophic phase of infection. Using immunocytochemistry, we found that ChELP2 is concentrated on the surface of bulbous biotrophic hyphae at the interface with living host cells but is absent from filamentous necrotrophic hyphae. We show that recombinant ChELP1 and ChELP2 bind chitin and chitin oligomers in vitro with high affinity and specificity and that both proteins suppress the chitin-triggered activation of two immune-related plant mitogen-activated protein kinases in the host Arabidopsis. Using RNAi-mediated gene silencing, we found that ChELP1 and ChELP2 are essential for fungal virulence and appressorium-mediated penetration of both Arabidopsis epidermal cells and cellophane membranes in vitro. The findings suggest a dual role for these LysM proteins as effectors for suppressing chitin-triggered immunity and as proteins required for appressorium function.


Assuntos
Arabidopsis/imunologia , Arabidopsis/microbiologia , Quitina/farmacologia , Colletotrichum/metabolismo , Espaço Extracelular/química , Proteínas Fúngicas/metabolismo , Imunidade Vegetal/efeitos dos fármacos , Sequência de Aminoácidos , Proteínas de Arabidopsis/metabolismo , Quitinases/metabolismo , Colletotrichum/efeitos dos fármacos , Colletotrichum/genética , Colletotrichum/patogenicidade , Proteínas Fúngicas/química , Regulação Fúngica da Expressão Gênica/efeitos dos fármacos , Genes Fúngicos , Hifas/metabolismo , Mutação/genética , Filogenia , Interferência de RNA , Transcrição Gênica/efeitos dos fármacos , Virulência/genética
5.
New Phytol ; 202(2): 485-498, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24372490

RESUMO

Nonrace specific disease resistance 1 (NDR1) is a conserved downstream regulator of resistance (R) protein-derived signaling. We identified two NDR1-like sequences (GmNDR1a, b) from soybean, and investigated their roles in R-mediated resistance and pathogen effector detection. Silencing GmNDR1a and b in soybean shows that these genes are required for resistance derived from the Rpg1-b, Rpg3, and Rpg4 loci, against Pseudomonas syringae (Psg) expressing avrB, avrB2 and avrD1, respectively. Immunoprecipitation assays show that the GmNDR1 proteins interact with the AvrB2 and AvrD1 Psg effectors. This correlates with the enhanced virulence of Psg avrB2 and Psg avrD1 in GmNDR1-silenced rpg3 rpg4 plants, even though these strains are not normally more virulent on plants lacking cognate R loci. The GmNDR1 proteins interact with GmRIN4 proteins, but not with AvrB, or its cognate R protein Rpg1-b. However, the GmNDR1 proteins promote AvrB-independent activation of Rpg1-b when coexpressed with a phosphomimic derivative of GmRIN4b. The role of GmNDR1 proteins in Rpg1-b activation, their direct interactions with AvrB2/AvrD1, and a putative role in the virulence activities of Avr effectors, provides the first experimental evidence in support of the proposed role for NDR1 in transducing extracellular pathogen-derived signals.


Assuntos
Proteínas de Bactérias/metabolismo , Resistência à Doença/genética , Glycine max/genética , Doenças das Plantas , Pseudomonas syringae/patogenicidade , Proteínas de Soja/genética , Fatores de Transcrição/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Regulação da Expressão Gênica de Plantas , Inativação Gênica , Genes de Plantas , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Pseudomonas syringae/genética , Pseudomonas syringae/metabolismo , Transdução de Sinais , Proteínas de Soja/metabolismo , Glycine max/metabolismo , Glycine max/microbiologia , Fatores de Transcrição/metabolismo , Virulência
6.
BMC Genomics ; 14: 658, 2013 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-24069909

RESUMO

BACKGROUND: Various bacteria can use non-ribosomal peptide synthesis (NRPS) to produce peptides or other small molecules. Conserved features within the NRPS machinery allow the type, and sometimes even the structure, of the synthesized polypeptide to be predicted. Thus, bacterial genome mining via in silico analyses of NRPS genes offers an attractive opportunity to uncover new bioactive non-ribosomally synthesized peptides. Xanthomonas is a large genus of Gram-negative bacteria that cause disease in hundreds of plant species. To date, the only known small molecule synthesized by NRPS in this genus is albicidin produced by Xanthomonas albilineans. This study aims to estimate the biosynthetic potential of Xanthomonas spp. by in silico analyses of NRPS genes with unknown function recently identified in the sequenced genomes of X. albilineans and related species of Xanthomonas. RESULTS: We performed in silico analyses of NRPS genes present in all published genome sequences of Xanthomonas spp., as well as in unpublished draft genome sequences of Xanthomonas oryzae pv. oryzae strain BAI3 and Xanthomonas spp. strain XaS3. These two latter strains, together with X. albilineans strain GPE PC73 and X. oryzae pv. oryzae strains X8-1A and X11-5A, possess novel NRPS gene clusters and share related NRPS-associated genes such as those required for the biosynthesis of non-proteinogenic amino acids or the secretion of peptides. In silico prediction of peptide structures according to NRPS architecture suggests eight different peptides, each specific to its producing strain. Interestingly, these eight peptides cannot be assigned to any known gene cluster or related to known compounds from natural product databases. PCR screening of a collection of 94 plant pathogenic bacteria indicates that these novel NRPS gene clusters are specific to the genus Xanthomonas and are also present in Xanthomonas translucens and X. oryzae pv. oryzicola. Further genome mining revealed other novel NRPS genes specific to X. oryzae pv. oryzicola or Xanthomonas sacchari. CONCLUSIONS: This study revealed the significant potential of the genus Xanthomonas to produce new non-ribosomally synthesized peptides. Interestingly, this biosynthetic potential seems to be specific to strains of Xanthomonas associated with monocotyledonous plants, suggesting a putative involvement of non-ribosomally synthesized peptides in plant-bacteria interactions.


Assuntos
Biologia Computacional/métodos , Genoma Bacteriano/genética , Biossíntese de Peptídeos Independentes de Ácido Nucleico/genética , Peptídeos/metabolismo , Xanthomonas/genética , Sequência de Aminoácidos , Simulação por Computador , Ácidos Graxos/biossíntese , Genes Bacterianos , Loci Gênicos/genética , Família Multigênica , Mapeamento Físico do Cromossomo , Plantas/microbiologia , Reação em Cadeia da Polimerase , Homologia de Sequência de Aminoácidos , Xanthomonas/enzimologia
7.
New Phytol ; 197(4): 1225-1235, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23278373

RESUMO

The Pseudomonas syringae effector AvrB interacts with four related soybean (Glycine max) proteins (GmRIN4a-d), three (GmRIN4b, c, d) of which also interact with the cognate resistance (R) protein, Rpg1-b. Here, we investigated the specific requirements for the GmRIN4 proteins in R-mediated resistance and examined the mechanism of Rpg1-b activation. Using virus-induced gene silencing, we show that only GmRIN4a and b are required for Rpg1-b-mediated resistance. In planta binding assays show that GmRIN4a can associate with Rpg1-b indirectly via GmRIN4b. Pathogen-delivered AvrB induces the phosphorylation of GmRIN4b alone, and prevents interactions between GmRIN4b and Rpg1-b or GmRIN4a. Consistent with this result, a phosphomimic derivative of GmRIN4b (pm4b) fails to bind Rpg1-b and GmRIN4a. Conversely, a phosphodeficient derivative of GmRIN4b (pd4b) continues to bind the R protein and GmRIN4a, in the presence of AvrB. Coexpression of Rpg1-b with pm4b, but not GmRIN4b or pd4b, induces cell death and ion leakage in the heterologous Nicotiana benthamiana. Our data suggest that the AvrB-induced phosphorylation of GmRIN4b, and the subsequent inhibition of interaction among GmRIN4b, GmRIN4a and Rpg1-b, might activate the R protein. Furthermore, even though GmRIN4c and d are not required for Rpg1-b-derived resistance, they do function in resistance derived from other R loci.


Assuntos
Resistência à Doença/genética , Glycine max/genética , Doenças das Plantas/imunologia , Proteínas de Plantas/fisiologia , Pseudomonas syringae/imunologia , Proteínas de Bactérias/metabolismo , Fosforilação , Doenças das Plantas/microbiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Glycine max/microbiologia
8.
Microbiology (Reading) ; 156(Pt 7): 1963-1974, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20378646

RESUMO

Type III secretion (T3S) systems play key roles in the assembly of flagella and the translocation of bacterial effector proteins into eukaryotic host cells. Eleven proteins which are conserved among gram-negative plant and animal pathogenic bacteria have been proposed to build up the basal structure of the T3S system, which spans both inner and outer bacterial membranes. We studied six conserved proteins, termed Hrc, predicted to reside in the inner membrane of the plant pathogen Xanthomonas campestris pv. vesicatoria. The membrane topology of HrcD, HrcR, HrcS, HrcT, HrcU and HrcV was studied by translational fusions to a dual alkaline phosphatase-beta-galactosidase reporter protein. Two proteins, HrcU and HrcV, were found to have the same membrane topology as the Yersinia homologues YscU and YscV. For HrcR, the membrane topology differed from the model for the homologue from Yersinia, YscR. For our data on three other protein families, exemplified by HrcD, HrcS and HrcT, we derived the first topology models. Our results provide what is believed to be the first complete model of the inner membrane topology of any bacterial T3S system and will aid in elucidating the architecture of T3S systems by ultrastructural analysis.


Assuntos
Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Membrana Celular/química , Membrana Celular/metabolismo , Xanthomonas vesicatoria/metabolismo , Proteínas de Bactérias/genética , Membrana Celular/genética , Doenças das Plantas/microbiologia , Estrutura Secundária de Proteína , Estrutura Terciária de Proteína , Transporte Proteico , Xanthomonas vesicatoria/química , Xanthomonas vesicatoria/genética
9.
Arch Cardiovasc Dis ; 113(1): 50-58, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31734140

RESUMO

BACKGROUND: International guidelines recommend that preoperative coronary angiography is performed on patients at risk of coronary disease who have infective endocarditis requiring surgical treatment. However, the risks of contrast-induced nephropathy or vegetation embolization in case of aortic endocarditis should be considered. AIMS: To assess the safety, therapeutic implications and prognostic impact of coronary angiography in patients requiring surgical treatment for active infective endocarditis. METHODS: This retrospective monocentric study was conducted in patients referred to a tertiary care centre for active endocarditis management with a theoretical indication for surgery between January 2013 and February 2017. RESULTS: One hundred and ninety-three patients were included; 73.1% were men, the mean age was 61.9±16.3 years and the median EuroSCORE II was 5.8%. One hundred and nineteen patients (61.7%) had aortic endocarditis, which was associated with aortic vegetation in 74 cases (38.3%). Invasive coronary angiography was performed in 142 patients (73.6%) - 130 (91.6%) by radial approach - and 14 patients were evaluated by coronary multislice computed tomography (one patient had exploration with both techniques). Acute renal failure after coronary angiography was observed in 15 patients (10.6%), two patients (1.4%) presented a stroke within 24h after coronary angiography, but none had aortic endocarditis. Among the 178 patients (92.2%) who underwent surgery, 35 (19.7%) had significant coronary lesion(s) and 25 (14.0%) underwent an associated coronary artery bypass graft. CONCLUSIONS: Preoperative coronary angiography in patients affected by infective endocarditis provides relevant information in a significant proportion of patients and can be performed safely.


Assuntos
Procedimentos Cirúrgicos Cardíacos , Angiografia por Tomografia Computadorizada , Angiografia Coronária , Doença da Artéria Coronariana/diagnóstico por imagem , Endocardite/cirurgia , Tomografia Computadorizada Multidetectores , Adulto , Idoso , Procedimentos Cirúrgicos Cardíacos/efeitos adversos , Angiografia por Tomografia Computadorizada/efeitos adversos , Angiografia Coronária/efeitos adversos , Doença da Artéria Coronariana/complicações , Doença da Artéria Coronariana/cirurgia , Endocardite/complicações , Endocardite/diagnóstico por imagem , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Tomografia Computadorizada Multidetectores/efeitos adversos , Valor Preditivo dos Testes , Estudos Retrospectivos , Medição de Risco , Fatores de Risco , Resultado do Tratamento
10.
FEMS Microbiol Lett ; 289(2): 225-32, 2008 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19054110

RESUMO

The filamentous ascomycete Botrytis cinerea is one of the most studied models for understanding the necrotrophic behaviour of phytopathogenic fungi. The genomes of two strains of B. cinerea have been sequenced (B05.10 and T4), which may contribute to elucidating the virulence polymorphism in this fungus. In this study, both strains were genetically modified in order to construct recipient strains designed to target genes that are hard to knock out. Deletions of BcKu70 gene in B05.10 strain and BcKu80 gene in T4 strain both affected the nonhomologous end-joining (NHEJ) DNA repair mechanism. NHEJ is responsible for the ectopic integration of gene replacement cassettes during fungal transformation and leads to a lower frequency of homologous recombination (HR). Ku deficiencies in B. cinerea did not disturb in vitro or in planta growth, but clearly improved HR efficiency for the putative sesquiterpene cyclase-encoding gene Cnd15, which was hard to knock out in a wild-type strain.


Assuntos
Botrytis/genética , Proteínas de Ligação a DNA/genética , Proteínas Fúngicas/genética , Técnicas de Inativação de Genes , Marcação de Genes , Botrytis/classificação , Botrytis/metabolismo , Proteínas de Ligação a DNA/metabolismo , Proteínas Fúngicas/metabolismo , Mutação , Filogenia , Doenças das Plantas/microbiologia , Recombinação Genética
11.
Diagnostics (Basel) ; 8(3)2018 Aug 18.
Artigo em Inglês | MEDLINE | ID: mdl-30126213

RESUMO

This paper presents the Wavelia microwave breast imaging system that has been recently installed at the Galway University Hospital, Ireland, for a first-in-human pilot clinical test. Microwave breast imaging has been extensively investigated over the last two decades as an alternative imaging modality that could potentially bring complementary information to state-of-the-art modalities such as X-ray mammography. Following an overview of the main working principles of this technology, the Wavelia imaging system architecture is presented, as are the radar signal processing algorithms that are used in forming the microwave images in which small tumors could be detectable for disease diagnosis. The methodology and specific quality metrics that have been developed to properly evaluate and validate the performance of the imaging system using complex breast phantoms that are scanned at controlled measurement conditions are also presented in the paper. Indicative results from the application of this methodology to the on-site validation of the imaging system after its installation at the hospital for pilot clinical testing are thoroughly presented and discussed. Given that the imaging system is still at the prototype level of development, a rigorous quality assessment and system validation at nominal operating conditions is very important in order to ensure high-quality clinical data collection.

12.
Front Plant Sci ; 9: 562, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29770142

RESUMO

The genome of the hemibiotrophic anthracnose fungus, Colletotrichum higginsianum, encodes a large inventory of putative secreted effector proteins that are sequentially expressed at different stages of plant infection, namely appressorium-mediated penetration, biotrophy and necrotrophy. However, the destinations to which these proteins are addressed inside plant cells are unknown. In the present study, we selected 61 putative effector genes that are highly induced in appressoria and/or biotrophic hyphae. We then used Agrobacterium-mediated transformation to transiently express them as N-terminal fusions with fluorescent proteins in cells of Nicotiana benthamiana for imaging by confocal microscopy. Plant compartments labeled by the fusion proteins in N. benthamiana were validated by co-localization with specific organelle markers, by transient expression of the proteins in the true host plant, Arabidopsis thaliana, and by transmission electron microscopy-immunogold labeling. Among those proteins for which specific subcellular localizations could be verified, nine were imported into plant nuclei, three were imported into the matrix of peroxisomes, three decorated cortical microtubule arrays and one labeled Golgi stacks. Two peroxisome-targeted proteins harbored canonical C-terminal tripeptide signals for peroxisome import via the PTS1 (peroxisomal targeting signal 1) pathway, and we showed that these signals are essential for their peroxisome localization. Our findings provide valuable information about which host processes are potentially manipulated by this pathogen, and also reveal plant peroxisomes, microtubules, and Golgi as novel targets for fungal effectors.

13.
ESC Heart Fail ; 4(2): 99-104, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28451445

RESUMO

AIMS: The main terminology used to describe heart failure (HF) is based on measurement of the left ventricular ejection fraction (LVEF). LVEF in the range of 40-49% was recently defined as HF with mid-range EF (HFmrEF) by the 2016 European Society of Cardiology guidelines. The purpose of our study was to assess the clinical profile and prognosis of patients with HF according to this new classification. METHODS AND RESULTS: A total of 482 patients referred for HF were retrospectively included over a period of 1 year. There were 258 (53%), 115 (24%), and 109 (23%) patients with HF with reduced EF (HFrEF), HFmrEF, and HF with preserved EF (HFpEF), respectively. Patient age increased, whereas left block bundle branch, brain natriuretic peptide level, and the use of beta-blocker and furosemide decreased from HFrEF to HFpEF. After adjustment for the age, patients with HFpEF and HFmrEF were more likely to have NYHA stage 2 dyspnea, had a higher systolic blood pressure, were less likely to have spironolactone, had lower furosemide dose, and had lower haemoglobin than those with HFrEF. Cardiovascular risk factors and medical history were similar in the three groups of patients. There was a 33% death rate after a mean follow-up of 32.2 ± 14.3 months. The survival was the same among patients whatever the group of HF (P = 0.884). CONCLUSIONS: Patients with HFrEF, HFmrEF, and HFpEF share the same cardiovascular risk factors, medical history, and prognosis. Patients with HFmrEF have a different clinical profile, which is nearly the same as patients with HFpEF, except for sex. These results question the relevance of this new classification of HF to stimulate research into this new group of patients.

14.
Amyloid ; 24(2): 101-109, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28553897

RESUMO

BACKGROUND: Among diagnosis associated with left ventricular hypertrophy (LVH), cardiac amyloidosis (CA) is a progressive disease with poor prognosis. Early noninvasive identification is of growing clinical importance. The objective of our study was to integrate clinical, biologic, electrocardiographic and echocardiographic parameters to build a diagnostic score in patients with LVH. METHODS AND RESULTS: One hundred and fourteen patients with LVH underwent a cardiac magnetic resonance (CMR) and a 99mTc-hydroxymethylene-diphosphonate scintigraphy (99mTc-HMDP) allowing to discriminate three groups of diagnoses: CA (n = 50 including 31, 18 and 1 ATTR, AL and AA amyloidosis), hypertrophic cardiomyopathy (n = 19) and unspecific cardiomyopathy (n = 45). Seven continuous variables associated with CA (systolic arterial pressure <130 mmHg; PR duration >200 ms; Sokolow index <12 mV; diastolic left ventricular posterior thickness >13 mm; E/Ea ratio >10; global longitudinal strain > -12% and sum of basal longitudinal strain > -47%) were selected and dichotomized according to the best cutoff value to build the diagnostic score, which was validated in an independent cohort of 34 patients with LVH from aortic stenosis. The area under the ROC curve for the diagnosis of CA using the score was 0.933 (95%CI 0.889-0.978). The best cut off value for the score was 3 leading to a sensitivity of 90% and specificity of 81%. Area under the ROC curve for the score was 0.932 in the validation cohort. A diagnostic score >3 was associated with a poorest prognosis. CONCLUSION: An integrated evaluation of 6 diagnostic factors including arterial blood pressure, ECG and echocardiographic parameters to build a diagnostic score is a simple and easily method to discriminate the 3 main CA in patients with LVH.


Assuntos
Amiloidose , Pressão Sanguínea , Eletrocardiografia , Imagem do Acúmulo Cardíaco de Comporta , Hipertrofia Ventricular Esquerda , Adulto , Idoso , Idoso de 80 Anos ou mais , Amiloidose/diagnóstico , Amiloidose/diagnóstico por imagem , Amiloidose/fisiopatologia , Feminino , Humanos , Hipertrofia Ventricular Esquerda/diagnóstico , Hipertrofia Ventricular Esquerda/diagnóstico por imagem , Hipertrofia Ventricular Esquerda/fisiopatologia , Masculino , Pessoa de Meia-Idade
15.
mBio ; 6(5): e01305-15, 2015 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-26374121

RESUMO

UNLABELLED: The cucumber anthracnose fungus Colletotrichum orbiculare forms specialized cells called appressoria for host penetration. We identified a gene, FAM1, encoding a novel peroxin protein that is essential for peroxisome biogenesis and that associates with Woronin bodies (WBs), dense-core vesicles found only in filamentous ascomycete fungi which function to maintain cellular integrity. The fam1 disrupted mutants were unable to grow on medium containing oleic acids as the sole carbon source and were nonpathogenic, being defective in both appressorium melanization and host penetration. Fluorescent proteins carrying peroxisomal targeting signals (PTSs) were not imported into the peroxisomes of fam1 mutants, suggesting that FAM1 is a novel peroxisomal biogenesis gene (peroxin). FAM1 did not show significant homology to any Saccharomyces cerevisiae peroxins but resembled conserved filamentous ascomycete-specific Pex22-like proteins which contain a predicted Pex4-binding site and are potentially involved in recycling PTS receptors from peroxisomes to the cytosol. C. orbiculare FAM1 complemented the peroxisomal matrix protein import defect of the S. cerevisiae pex22 mutant. Confocal microscopy of Fam1-GFP (green fluorescent protein) fusion proteins and immunoelectron microscopy with anti-Fam1 antibodies showed that Fam1 localized to nascent WBs budding from peroxisomes and mature WBs. Association of Fam1 with WBs was confirmed by colocalization with WB matrix protein CoHex1 (C. orbiculare Hex1) and WB membrane protein CoWsc (C. orbiculare Wsc) and by subcellular fractionation and Western blotting with antibodies to Fam1 and CoHex1. In WB-deficient cohex1 mutants, Fam1 was redirected to the peroxisome membrane. Our results show that Fam1 is a WB-associated peroxin required for pathogenesis and raise the possibility that localized receptor recycling occurs in WBs. IMPORTANCE: Colletotrichum orbiculare is a fungus causing damaging disease on Cucurbitaceae plants. In this paper, we characterize a novel peroxisome biogenesis gene from this pathogen called FAM1. Although no genes with significant homology are present in Saccharomyces cerevisiae, FAM1 contains a predicted Pex4-binding site typical of Pex22 proteins, which function in the recycling of PTS receptors from peroxisomes to the cytosol. We show that FAM1 complements the defect in peroxisomal matrix protein import of S. cerevisiae pex22 mutants and that fam1 mutants are completely defective in peroxisome function, fatty acid metabolism, and pathogenicity. Remarkably, we found that this novel peroxin is specifically localized on the bounding membrane of Woronin bodies, which are small peroxisome-derived organelles unique to filamentous ascomycete fungi that function in septal pore plugging. Our finding suggests that these fungi have coopted the Woronin body for localized receptor recycling during matrix protein import.


Assuntos
Colletotrichum/enzimologia , Cucurbitaceae/microbiologia , Proteínas Fúngicas/metabolismo , Biogênese de Organelas , Peroxissomos/metabolismo , Doenças das Plantas/microbiologia , Fatores de Virulência/metabolismo , Colletotrichum/genética , Proteínas Fúngicas/genética , Deleção de Genes , Teste de Complementação Genética , Proteínas de Membrana/deficiência , Microscopia Confocal , Microscopia Imunoeletrônica , Organelas/química , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae , Fatores de Virulência/genética
16.
J Proteomics ; 97: 256-64, 2014 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-23603630

RESUMO

Pathogenicity of the rice pathogenic bacterium Xanthomonas oryzae pv. oryzae depends on a Hrp (hypersensitive response and pathogenicity) type III secretion system; the expression of which is induced in planta. Expression of the hrp operons is under transcriptional control of two key regulatory proteins, HrpG and HrpX. To identify new proteins that are co-regulated with the type III secretion system, we employed comparative proteomics. Cells of X. oryzae pv. oryzae ectopically expressing hrpX were compared to wild-type cells grown in vitro. Twenty protein spots with different abundances in both samples were identified by 2D-DIGE and LC-MS/MS. Seven spots could be unambiguously identified, corresponding to the HrpB1 protein, two different peptidyl-prolyl cis-trans isomerases, a component of an ATP binding cassette (ABC) transport system, an adenylate kinase, and a secreted protein of unknown function. Interestingly, the isoelectric point of the adenylate kinase was found to be under control of HrpX, most likely due to post-translational modification. Indeed, two glutamate residues of the adenylate kinase were found to be methylated but this modification did not account for the shift in electrophoretic mobility. In summary, we identified new HrpX-regulated proteins of X. oryzae pv. oryzae that might be important for pathogenicity. This article is part of a Special Issue entitled: Trends in microbial proteomics. BIOLOGICAL SIGNIFICANCE: We use 2D-DIGE to compare the proteomes of rice-pathogenic xanthomonads. We identify seven proteins that are co-regulated with the type III secretion system. We find post-translational glutamate methylation of a bacterial adenylate cyclase. The newly identified HrpX-regulated proteins might be important for pathogenicity.


Assuntos
Proteínas de Bactérias/metabolismo , Óperon/fisiologia , Proteômica , Fatores de Transcrição/metabolismo , Xanthomonas/metabolismo , Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/metabolismo , Proteínas de Bactérias/genética , Oryza/microbiologia , Peptidilprolil Isomerase/genética , Peptidilprolil Isomerase/metabolismo , Doenças das Plantas/microbiologia , Fatores de Transcrição/genética , Xanthomonas/genética , Xanthomonas/patogenicidade
17.
Curr Opin Plant Biol ; 16(4): 527-33, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23870750

RESUMO

Systemic acquired resistance (SAR) is a type of pathogen-induced broad-spectrum resistance in plants. During SAR, primary infection-induced rapid generation and transportation of mobile signal(s) 'prepare' the rest of the plant for subsequent infections. Several, seemingly unrelated, mobile chemical inducers of SAR have been identified, at least two of which function in a feed-back regulatory loop with a lipid transfer-like protein. Signal(s) perception in the systemic tissues relies on the presence of an intact cuticle, the waxy layer covering all aerial parts of the plant. SAR results in chromatin modifications, which prime systemic tissues for enhanced and rapid signaling derived from salicylic acid, which along with its signaling components is key for SAR induction. This review summarizes recent findings related to SAR signal generation, movement, and perception.


Assuntos
Resistência à Doença , Regulação da Expressão Gênica de Plantas , Doenças das Plantas/genética , Plantas/genética , Transdução de Sinais , Cromatina , Imunidade Inata , Doenças das Plantas/microbiologia , Plantas/imunologia , Ácido Salicílico/metabolismo
18.
Ultrasonics ; 52(5): 622-7, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22325847

RESUMO

In many industrial processes where online control is necessary such as in the food industry, the real time monitoring of visco-elastic properties is essential to ensure the quantity of production. Acoustic methods have shown that reliable properties could be obtained from measurements of velocity and attenuation. This paper proposes a simple, real time ultrasound method for monitoring linear medium properties (phase velocity and attenuation) that vary in time. The method is based on a pulse echo measurement and is self-calibrated. Results on a silica gel are reported and the importance of taking into account the changes of the mechanical loading on the front face of the transducer will be shown. This is done through a modification of the emission and reception transfer parameters. The simultaneous measurement of the input and output currents and voltages enables these parameters to be calculated during the reaction. The variations of the transfer parameters are in the order of 6% and predominate other effects. The evolution of the ultrasonic longitudinal wave phase velocity and attenuation as a function of time allows the characteristic times of the chemical reaction to be determined. The results are well correlated with the gelation time measured by rheological method at low frequency.


Assuntos
Géis/química , Reologia/métodos , Dióxido de Silício/química , Ultrassom , Algoritmos , Calibragem , Elasticidade , Transição de Fase , Controle de Qualidade , Transdutores , Vibração , Viscosidade
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