RESUMO
Geminiviruses are DNA viruses that cause severe crop losses in different parts of the world, and there is a need for genetic sources of resistance to help combat them. Arabidopsis has been used as a source for virus-resistant genes that derive from alterations in essential host factors. We used a virus-induced gene silencing (VIGS) vector derived from the geminivirus Cabbage leaf curl virus (CaLCuV) to assess natural variation in virus-host interactions in 190 Arabidopsis accessions. Silencing of CH-42, encoding a protein needed to make chlorophyll, was used as a visible marker to discriminate asymptomatic accessions from those showing resistance. There was a wide range in symptom severity and extent of silencing in different accessions, but two correlations could be made. Lines with severe symptoms uniformly lacked extensive VIGS, and lines that showed attenuated symptoms over time (recovery) showed a concomitant increase in the extent of VIGS. One accession, Pla-1, lacked both symptoms and silencing, and was immune to wild-type infectious clones corresponding to CaLCuV or Beet curly top virus (BCTV), which are classified in different genera in the Geminiviridae. It also showed resistance to the agronomically important Tomato yellow leaf curl virus (TYLCV). Quantitative trait locus mapping of a Pla-1 X Col-0 F2 population was used to detect a major peak on chromosome 1, which is designated gip-1 (geminivirus immunity Pla-1-1). The recessive nature of resistance to CaLCuV and the lack of obvious candidate genes near the gip-1 locus suggest that a novel resistance gene(s) confers immunity.
Assuntos
Arabidopsis/virologia , Geminiviridae/imunologia , Doenças das Plantas/virologia , Imunidade Vegetal , Inativação Gênica , Doenças das Plantas/imunologia , Locos de Características Quantitativas/genéticaRESUMO
Standard computer vision methods are usually based on powerful contact-less measurement approaches but applications, especially at the micro-scale, are restricted by finite depth-of-field and fixed working distance of imaging devices. Digital holography is a lensless, indirect imaging method recording the optical wave diffracted by the object onto the image sensor. The object is reconstructed numerically by propagating the recorded wavefront backward. The object distance becomes a computation parameter that can be chosen arbitrarily and adjusted to match the object position. No refractive lens is used and usual depth-of-field and working distance limitations are replaced by less restrictive ones tied to the laser-source coherence-length and to the size and resolution of the camera sensor. This paper applies digital holography to artificial visual in-plane position sensing with an extra-large range-to-resolution ratio. The object is made of a pseudoperiodic pattern allowing a subpixel resolution as well as a supra field-of-observation displacement range. We demonstrate an in-plane resolution of 50 nm and 0.002deg. in X, Y and θ respectively, over a working distance range of more than 15 cm. The allowed workspace extends over 12×10×150mm3. Digital holography extends the field of application of computer vision by allowing an extra-large range of working distances inaccessible to refractive imaging systems.
RESUMO
A decade of studies on middle cortex (MC) formation in the root endodermis of Arabidopsis (Arabidopsis thaliana) have revealed a complex regulatory network that is orchestrated by several GRAS family transcription factors, including SHORT-ROOT (SHR), SCARECROW (SCR), and SCARECROW-LIKE3 (SCL3). However, how their functions are regulated remains obscure. Here we show that mutations in the SEUSS (SEU) gene led to a higher frequency of MC formation. seu mutants had strongly reduced expression of SHR, SCR, and SCL3, suggesting that SEU positively regulates these genes. Our results further indicate that SEU physically associates with upstream regulatory sequences of SHR, SCR, and SCL3; and that SEU has distinct genetic interactions with these genes in the control of MC formation, with SCL3 being epistatic to SEU. Similar to SCL3, SEU was repressed by the phytohormone GA and induced by the GA biosynthesis inhibitor paclobutrazol, suggesting that SEU acts downstream of GA signaling to regulate MC formation. Consistently, we found that SEU mediates the regulation of SCL3 by GA signaling. Together, our study identifies SEU as a new critical player that integrates GA signaling with transcriptional inputs from the SHR-SCR-SCL3 module to regulate MC formation in the Arabidopsis root.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Proteínas Correpressoras/metabolismo , Giberelinas/metabolismo , Raízes de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Arabidopsis/citologia , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas Correpressoras/genética , Epistasia Genética , Regulação da Expressão Gênica de Plantas , Microscopia Confocal , Modelos Genéticos , Mutação , Raízes de Plantas/citologia , Raízes de Plantas/genética , Plantas Geneticamente Modificadas , Ligação Proteica , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais , Fatores de Transcrição/genética , Técnicas do Sistema de Duplo-HíbridoRESUMO
Plant meristems, like animal stem cell niches, maintain a pool of multipotent, undifferentiated cells that divide and differentiate to give rise to organs. In Arabidopsis (Arabidopsis thaliana), the carpel margin meristem is a vital meristematic structure that generates ovules from the medial domain of the gynoecium, the female floral reproductive structure. The molecular mechanisms that specify this meristematic region and regulate its organogenic potential are poorly understood. Here, we present a novel approach to analyze the transcriptional signature of the medial domain of the Arabidopsis gynoecium, highlighting the developmental stages that immediately proceed ovule initiation, the earliest stages of seed development. Using a floral synchronization system and a SHATTERPROOF2 (SHP2) domain-specific reporter, paired with FACS and RNA sequencing, we assayed the transcriptome of the gynoecial medial domain with temporal and spatial precision. This analysis reveals a set of genes that are differentially expressed within the SHP2 expression domain, including genes that have been shown previously to function during the development of medial domain-derived structures, including the ovules, thus validating our approach. Global analyses of the transcriptomic data set indicate a similarity of the pSHP2-expressing cell population to previously characterized meristematic domains, further supporting the meristematic nature of this gynoecial tissue. Our method identifies additional genes including novel isoforms, cis-natural antisense transcripts, and a previously unrecognized member of the REPRODUCTIVE MERISTEM family of transcriptional regulators that are potential novel regulators of medial domain development. This data set provides genome-wide transcriptional insight into the development of the carpel margin meristem in Arabidopsis.
Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Proteínas de Domínio MADS/genética , Meristema/genética , Transcriptoma , Arabidopsis/anatomia & histologia , Proteínas de Arabidopsis/isolamento & purificação , Sequência de Bases , Hidrato de Cloral , DNA Antissenso , Flores/genética , Genoma de Planta , Hibridização In Situ , Ácidos Indolacéticos/farmacologia , Proteínas de Domínio MADS/isolamento & purificação , Meristema/crescimento & desenvolvimento , Meristema/metabolismo , Microscopia Confocal , Óvulo Vegetal/citologia , Óvulo Vegetal/crescimento & desenvolvimento , Óvulo Vegetal/metabolismo , Isoformas de Proteínas , Protoplastos , RNA de Plantas/química , RNA de Plantas/isolamento & purificação , Sementes/crescimento & desenvolvimento , Alinhamento de Sequência , Fatores de Transcrição , Ativação TranscricionalRESUMO
Scaffold or matrix attachment regions (S/MARs) are found in all eukaryotes. The pattern of distribution and genomic context of S/MARs is thought to be important for processes such as chromatin organization and modulation of gene expression. Despite the importance of such processes, much is unknown about the large-scale distribution and sequence content of S/MARs in vivo. Here, we report the use of tiling microarrays to map 1358 S/MARs on Arabidopsis thaliana chromosome 4 (chr4). S/MARs occur throughout chr4, spaced much more closely than in the large plant and animal genomes that have been studied to date. Arabidopsis S/MARs can be divided into five clusters based on their association with other genomic features, suggesting a diversity of functions. While some Arabidopsis S/MARs may define structural domains, most occur near the transcription start sites of genes. Genes associated with these S/MARs have an increased probability of expression, which is particularly pronounced in the case of transcription factor genes. Analysis of sequence motifs and 6-mer enrichment patterns show that S/MARs are preferentially enriched in poly(dA:dT) tracts, sequences that resist nucleosome formation, and the majority of S/MARs contain at least one nucleosome-depleted region. This global view of S/MARs provides a framework to begin evaluating genome-scale models for S/MAR function.
Assuntos
Proteínas de Arabidopsis/fisiologia , Arabidopsis/genética , Regiões de Interação com a Matriz , Nucleossomos/metabolismo , Poli dA-dT/metabolismo , Fatores de Transcrição/fisiologia , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Cromatina/metabolismo , Regulação da Expressão Gênica de Plantas , Motivos de Nucleotídeos , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
BACKGROUND AND OBJECTIVES: Blood-based biomarkers have emerged as minimally invasive options for evaluating cognitive impairment. Most studies to date have assessed them in research cohorts, limiting their generalization to everyday clinical practice. We evaluated their diagnostic performance and clinical applicability in a prospective, real-world, memory clinic cohort. METHODS: All patients referred with suspected cognitive impairment between July 2019 and June 2021 were prospectively invited to participate. Five plasma biomarkers (tau phosphorylated at threonine 181 [p-tau181], glial fibrillary acidic protein [GFAP], neurofilament light chain [NfL], total tau [t-tau], and ubiquitin C-terminal hydrolase L1 [UCH-L1]) were determined with single-molecule array. Performance was assessed in comparison to clinical diagnosis (blinded to plasma results) and amyloid status (CSF/PET). A group of cognitively unimpaired (CU) controls was also included. RESULTS: Three hundred forty-nine participants (mean age 68, SD 8.3 years) and 36 CU controls (mean age 61.7, SD 8.2 years) were included. In the subcohort with available Alzheimer disease (AD) biomarkers (n = 268), plasma p-tau181 and GFAP had a high diagnostic accuracy to differentiate AD from non-neurodegenerative causes (area under the receiver operating characteristic curve 0.94 and 0.92, respectively), with p-tau181 systematically outperforming GFAP. Plasma p-tau181 levels predicted amyloid status (85% sensitivity and specificity) with accurate individual prediction in approximately 60% of the patients. Plasma NfL differentiated frontotemporal dementia (FTD) syndromes from CU (0.90) and non-neurodegenerative causes (0.93), whereas the discriminative capacity with AD and between all neurodegenerative and non-neurodegenerative causes was less accurate. A combination of p-tau181 and NfL identified FTD with 82% sensitivity and 85% specificity and had a negative predictive value for neurodegenerative diagnosis of 86%, ruling out half of the non-neurodegenerative diagnoses. In the subcohort without AD biomarkers, similar results were obtained. T-tau and UCH-L1 did not offer added diagnostic value. DISCUSSION: Plasma p-tau181 predicted amyloid status with high accuracy and could have potentially avoided CSF/amyloid PET testing in approximately 60% of subjects in a memory clinic setting. NfL was useful for identifying FTD from non-neurodegenerative causes but behaved worse than p-tau181 in all other comparisons. Combining p-tau181 and NfL improved diagnostic performance for FTD and non-neurodegenerative diagnoses. However, the 14% false-negative results suggest that further improvement is needed before implementation outside memory clinics. CLASSIFICATION OF EVIDENCE: This study provides Class I evidence that plasma p-tau181 correlates with the presence or absence of AD and a combination of plasma p-tau181 and NfL correlates moderately well with a diagnosis of FTD.
Assuntos
Doença de Alzheimer , Demência Frontotemporal , Memória Episódica , Doença de Pick , Humanos , Idoso , Pessoa de Meia-Idade , Proteínas tau , Demência Frontotemporal/diagnóstico , Peptídeos beta-Amiloides , Doença de Alzheimer/psicologia , BiomarcadoresRESUMO
Sporadic early-onset Alzheimer's disease (EOAD) and autosomal dominant Alzheimer's disease (ADAD) provide the opportunity to investigate the physiopathological mechanisms in the absence of aging, present in late-onset forms. Frontotemporal dementia (FTD) causes early-onset dementia associated to tau or TDP43 protein deposits. A 15% of FTD cases are caused by mutations in C9orf72, GRN, or MAPT genes. Lymphoblastoid cell lines (LCLs) have been proposed as an alternative to brain tissue for studying earlier phases of neurodegenerative diseases. The aim of this study is to investigate the expression profile in EOAD, ADAD, and sporadic and genetic FTD (sFTD and gFTD, respectively), using brain tissue and LCLs. Sixty subjects of the following groups were included: EOAD, ADAD, sFTD, gFTD, and controls. Gene expression was analyzed with Clariom D microarray (Affymetrix). Brain tissue pairwise comparisons revealed six common differentially expressed genes (DEG) for all the patients' groups compared with controls: RGS20, WIF1, HSPB1, EMP3, S100A11 and GFAP. Common up-regulated biological pathways were identified both in brain and LCLs (including inflammation and glial cell differentiation), while down-regulated pathways were detected mainly in brain tissue (including synaptic signaling, metabolism and mitochondrial dysfunction). CD163, ADAMTS9 and LIN7A gene expression disruption was validated by qPCR in brain tissue and NrCAM in LCLs in their respective group comparisons. In conclusion, our study highlights neuroinflammation, metabolism and synaptic signaling disturbances as common altered pathways in different AD and FTD forms. The use of LCLs might be appropriate for studying early immune system and inflammation, and some neural features in neurodegenerative dementias.
Assuntos
Doença de Alzheimer , Demência Frontotemporal , Doença de Alzheimer/genética , Doença de Alzheimer/patologia , Encéfalo/patologia , Proteína C9orf72/genética , Linhagem Celular , Demência Frontotemporal/genética , Demência Frontotemporal/patologia , Expressão Gênica , Humanos , Inflamação/patologia , Glicoproteínas de Membrana/genética , Proteínas de Membrana/genética , Mutação , Proteínas de Transporte Vesicular , Proteínas tau/genéticaRESUMO
BACKGROUND: Early onset Alzheimer's disease (EOAD) represents a diagnostic challenge and is associated with a high diagnostic delay and misdiagnosis. OBJECTIVE: To describe clinical and pathological data from a pathologically confirmed EOAD cohort and evaluate evolving trends in clinical-pathological correlation accuracy. METHODS: Retrospective review of clinical and neuropathological data of pathologically confirmed EOAD patients (age at onset [AAO]â<â60). Comparison between two periods: 1994- 2009 and 2010- 2018. RESULTS: Eighty brain donors were included. Mean AAO, age at death, and diagnostic delay was 55, 66, and 3 years, respectively. Twenty-nine percent had a nonamnestic presentation. Sixteen percent were given a non-AD initial clinical diagnosis (initial misdiagnosis) and 14% received a final misdiagnosis. Nonamnestic presentation patients received more misdiagnoses than amnestic presentation ones (39% versus 7% and 39% versus 3.5%, on initial and final misdiagnosis, respectively). When comparing both time periods, a trend towards a higher diagnostic accuracy in the 2010- 2018 period was observed, mainly on initial misdiagnosis in nonamnestic presentation patients (53% versus 13%, pâ=â0.069). Diagnostic delay was similar between both periods. Cerebral amyloid angiopathy (96%) and Lewy body co-pathology (55%) were very frequent, while limbic-predominant age-related TDP-43 encephalopathy pathologic changes were only present in 12.5%. CONCLUSION: In the last decade, there has been a trend towards improved diagnostic accuracy in EOAD, which might be explained by improved diagnostic criteria, increasing experience on EOAD and the beginning of the use of biomarkers, although diagnostic delay remains similar. Concomitant neuropathology was very frequent despite the relatively young age of brain donors.
Assuntos
Doença de Alzheimer , Idade de Início , Doença de Alzheimer/diagnóstico , Doença de Alzheimer/patologia , Encéfalo/diagnóstico por imagem , Encéfalo/patologia , Diagnóstico Tardio , Humanos , Corpos de Lewy/patologiaRESUMO
The double fertilization of the female gametophyte initiates embryogenesis and endosperm development in seeds via the activation of genes involved in cell differentiation, organ patterning, and growth. A subset of genes expressed in endosperm exhibit imprinted expression, and the correct balance of gene expression between parental alleles is critical for proper endosperm and seed development. We use a transcriptional time series analysis to identify genes that are associated with key shifts in seed development, including genes associated with secondary cell wall synthesis, mitotic cell cycle, chromatin organization, auxin synthesis, fatty acid metabolism, and seed maturation. We relate these genes to morphological changes in Mimulus seeds. We also identify four endosperm-expressed transcripts that display imprinted (paternal) expression bias. The imprinted status of these four genes is conserved in other flowering plants, suggesting that they are functionally important in endosperm development. Our study explores gene regulatory dynamics in a species with ab initio cellular endosperm development, broadening the taxonomic focus of the literature on gene expression in seeds. Moreover, it is the first to validate genes with imprinted endosperm expression in Mimulus guttatus, and will inform future studies on the genetic causes of seed failure in this model system.
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Estas reflexões fazem parte de uma pesquisa realizada sobre questões patrimoniais e históricas de uma cidade emblemática gaúcha que procura mostrar no debate sobre a sua colonização algum aspecto das imagens de negação étnica ocorrida no processo de assimilação da população riograndense. Nesse sentido, as vivências cotidianas acontecidas nas cidades pioneiras servem para repensar o sentido figurativo da dimensão heróico-mítica dos pampas-farrapos como mostra nosso estudo de caso da vida social na cidade de Piratini - RS no tempo da colonização até os dias de hoje
Assuntos
Etnicidade , AculturaçãoRESUMO
Existen evidencias que relacionan hiperinsulinemia con aterosclerosis, pero no hay consenso sobre la fisiopatogenia de esta asociación. Está reconocido que el aumento del espesor intima media arterial ocurre en las etapas iniciales de la aterosclerosis. Objetivo: 1) Relacionar el nivel de insulina en sangre con el espesor intima media (EIM) de la pared arterial en dos grupos de sujetos jóvenes no diabéticos, Hiperinsulinémicos y Normoinsulinémicos. 2) Analizar la asociación entre el Homeostasis Model Assessmen (HOMA) y el EIM de la pared arterial en ambos grupos. 3) Comparar el colesterol total (col-total), colesterol-HDL (col-HDL), colesterol-LDL (col-LDL), triglicéridos (TG), índice cintura cadera (ICC), presión arterial (PA), índice de masa corporal (IMC) y ácido úrico, entre ambos grupos investigados y su asociación con el EIM de la pared arterial. Se estudiaron 98 sujetos de ambos sexos con edades entre 18 y 35 años, 59 Hiperinsulinémicos y 39 Normoinsulinémicos, grupo control (GC). Se determinó en ayunas: Insulinemia, glucemia, col-total, col-HDL, col-LDL, TG, ácido úrico y HOMA. Luego del suministro por vía oral de 75 grs. de glucosa, se controló glucemia e insulinemia a los 30, 60, y 120 minutos. Se valoró la PA, el ICC y el IMC. Se midió el EIM en el segmento de la arteria carótida común (ACC), sobre la pared ecográficamente lejana, utilizando ultrasonido de superficie con un transductor de 7.5 MHZ El grupo hiperinsulinémico presentó diferencias significativas con respecto al normoinsulinémico (GC) en todas las variables muestreadas.