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1.
JBI Evid Synth ; 21(4): 753-761, 2023 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-36214503

RESUMO

OBJECTIVE: The objective of this review is to identify and characterize the use of the natural environment/outdoor space by occupational therapists working in mental health care. INTRODUCTION: Research has shown that the natural environment is beneficial for our health and can be used to help people who are experiencing mental health problems. Occupational therapists are well placed to assess and utilize the environment when treating people with mental health problems; however, the use of the natural environment/outdoor space by occupational therapists working in mental health is unclear. INCLUSION CRITERIA: This scoping review will include both primary research and gray literature relating to the use of the natural environment/outdoor space in mental health occupational therapy practice. The review will be limited to studies published in English. There will be no geographical or age restrictions. METHODS: Embase (Ovid), MEDLINE (Ovid), CINAHL (EBSCO), PsycINFO (ProQuest), AMED (EBSCO), Trip Database, Emcare (Ovid), and OTSeeker will be searched for studies. Unpublished studies and gray literature will be searched using GreyNet and National Grey Literature Collection, alongside professional magazines and websites. Titles and abstracts will be screened by 2 independent reviewers for assessment against the inclusion criteria, followed by a full-text review and data extraction. Any disagreements will be discussed with a third reviewer. Data will be extracted using a data extraction tool developed by the reviewers, and presented in tabular format, accompanied by a narrative summary describing how the results relate to the review objective and question.


Assuntos
Saúde Mental , Terapeutas Ocupacionais , Humanos , Meio Ambiente , Literatura de Revisão como Assunto
2.
J Med Internet Res ; 23(12): e26767, 2021 12 20.
Artigo em Inglês | MEDLINE | ID: mdl-34932010

RESUMO

BACKGROUND: The use of smart speakers to improve well-being had been trialed in social care by others; however, we were not aware of their implementation in most care homes across a region in the Southwest of the United Kingdom. For the widespread adoption of new technology, it must be locally demonstrable and become normalized. OBJECTIVE: The aim of this study was to install smart speakers in care homes in a rural and coastal region and to explore if and how the devices were being used, the barriers to their implementation, and their potential benefits. METHODS: Email, workshops, drop-in sessions, phone, and cold calling was used to contact all 230 care homes, offering a free smart speaker and some advisory support. Care homes accepting the devices were asked to complete a feedback diary. Nonresponse rate for diary completion was high and was thus supplemented with a telephone survey. RESULTS: Over the course of 7 months, we installed 156 devices in 92 care homes for older people, 50 devices for people with physical or mental health needs, and 8 for others. The devices were used mainly for music but also for poetry, recipes, light controls, jokes, and video calls. Care home managers reported the benefits for the residents, including enhanced engagement with home activities, enjoyment, calming effects, and the acquisition of new skills. Implementation problems included internet connectivity, staff capacity, and skills. CONCLUSIONS: Affordable consumer devices such as smart speakers should be installed in all care homes to benefit residents. Voice-activated technologies are easy to use and promote interaction. This study indicates that implementation in care homes was possible and that smart speakers had multifaceted benefits for residents and staff. Most care homes in this region now use smart speakers for their residents, thereby normalizing this practice.


Assuntos
Ansiedade , Tecnologia , Idoso , Humanos , Reino Unido
3.
PLoS One ; 11(10): e0165448, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27798702

RESUMO

Rising global temperatures may increase the rates of soil organic matter decomposition by heterotrophic microorganisms, potentially accelerating climate change further by releasing additional carbon dioxide (CO2) to the atmosphere. However, the possibility that microbial community responses to prolonged warming may modify the temperature sensitivity of soil respiration creates large uncertainty in the strength of this positive feedback. Both compensatory responses (decreasing temperature sensitivity of soil respiration in the long-term) and enhancing responses (increasing temperature sensitivity) have been reported, but the mechanisms underlying these responses are poorly understood. In this study, microbial biomass, community structure and the activities of dehydrogenase and ß-glucosidase enzymes were determined for 18 soils that had previously demonstrated either no response or varying magnitude of enhancing or compensatory responses of temperature sensitivity of heterotrophic microbial respiration to prolonged cooling. The soil cooling approach, in contrast to warming experiments, discriminates between microbial community responses and the consequences of substrate depletion, by minimising changes in substrate availability. The initial microbial community composition, determined by molecular analysis of soils showing contrasting respiration responses to cooling, provided evidence that the magnitude of enhancing responses was partly related to microbial community composition. There was also evidence that higher relative abundance of saprophytic Basidiomycota may explain the compensatory response observed in one soil, but neither microbial biomass nor enzymatic capacity were significantly affected by cooling. Our findings emphasise the key importance of soil microbial community responses for feedbacks to global change, but also highlight important areas where our understanding remains limited.


Assuntos
Microbiota , Solo/química , Temperatura , Biomassa , Respiração Celular , Fumigação , Modelos Lineares , Análise de Sequência de DNA , Fatores de Tempo
4.
Sci Rep ; 6: 19335, 2016 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-26757823

RESUMO

Tree disease epidemics are a global problem, impacting food security, biodiversity and national economies. The potential for conservation and breeding in trees is hampered by complex genomes and long lifecycles, with most species lacking genomic resources. The European Ash tree Fraxinus excelsior is being devastated by the fungal pathogen Hymenoscyphus fraxineus, which causes ash dieback disease. Taking this system as an example and utilizing Associative Transcriptomics for the first time in a plant pathology study, we discovered gene sequence and gene expression variants across a genetic diversity panel scored for disease symptoms and identified markers strongly associated with canopy damage in infected trees. Using these markers we predicted phenotypes in a test panel of unrelated trees, successfully identifying individuals with a low level of susceptibility to the disease. Co-expression analysis suggested that pre-priming of defence responses may underlie reduced susceptibility to ash dieback.


Assuntos
Adaptação Biológica/genética , Fraxinus/genética , Fraxinus/microbiologia , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Transcriptoma , Biomarcadores , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Polimorfismo de Nucleotídeo Único
5.
Data Brief ; 3: 51-5, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26217717

RESUMO

This data article includes SNP scoring across lines of the Brassica napus TNDH population based on Illumina sequencing of mRNA, expanded to 75 lines. The 21, 323 mapped markers defined 887 recombination bins, representing an updated genetic linkage map for the species. Based on this new map, 5 genome sequence scaffolds were split and the order and orientation of scaffolds updated to establish a new pseudomolecule specification. The order of unigenes and SNP array probes within these pseudomolecules was determined. Unigenes were assessed for sequence similarity to the A and C genomes. The 57, 246 that mapped to both enabled the collinearity of the A and C genomes to be illustrated graphically. Although the great majority was in collinear positions, some were not. Analyses of 60 such instances are presented, suggesting that the breakdown in collinearity was largely due to either the absence of the homoeologue on one genome (resulting in sequence match to a paralogue) or multiple similar sequences being present. The mRNAseq datasets for the TNDH lines are available from the SRA repository (ERA283648); the remaining datasets are supplied with this article.

6.
BMC Evol Biol ; 15: 22, 2015 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-25879701

RESUMO

BACKGROUND: The A Disintegrin-like and Metalloproteinase domain with Thrombospondin-1 motifs (ADAMTS) enzymes comprise 19 mammalian zinc-dependent metalloproteinases (metzincins) with homologues in a wide range of invertebrates. ADAMTS enzymes have a broad range of functions in development and diseases due to their extracellular matrix remodelling activity. Here, we report a detailed characterisation of their evolutionary conservation across vertebrates. RESULTS: Using bioinformatics complemented with de novo sequencing, gene sequences for ADAMTS enzymes were obtained from a variety of organisms. Detailed evolutionary analyses revealed a high level of conservation across vertebrates with evidence of ADAMTS gene expansion during two rounds of whole genome duplication (WGD) in vertebrates, while tandem duplication events and gene loss were also apparent. However, the additional round of teleost-specific WGD did not have a significant effect on ADAMTS gene family members suggesting their conserved roles have remained constant in teleost fish. Quantitative reverse-transcriptase polymerase chain reaction analysis revealed dynamic expression of adamts genes throughout zebrafish embryonic development reflecting the key conserved roles they play in vertebrate embryogenesis. Notably, several adamts mRNAs were maternally expressed with a dramatic increase in mRNA levels coinciding with zygotic expression and organogenesis. Broad adamts mRNA expression was also demonstrated in several adult organs indicating potential roles in adult homeostasis. CONCLUSIONS: Our data highlight the evolution of the ADAMTS gene family through duplication processes across metazoans supplemented by a burst of amplification through vertebrate WGD events. It also strongly posits the zebrafish as a potential model species to further elucidate the function of ADAMTS enzymes during vertebrate development.


Assuntos
Evolução Molecular , Metaloendopeptidases/química , Metaloendopeptidases/genética , Proteínas de Peixe-Zebra/química , Proteínas de Peixe-Zebra/genética , Peixe-Zebra/genética , Proteínas ADAM/química , Proteínas ADAM/genética , Proteínas ADAM/metabolismo , Animais , Duplicação Gênica , Regulação da Expressão Gênica no Desenvolvimento , Genoma , Metaloendopeptidases/metabolismo , Filogenia , Estrutura Terciária de Proteína , Vertebrados/genética , Peixe-Zebra/embriologia , Proteínas de Peixe-Zebra/metabolismo
7.
DNA Res ; 21(6): 613-25, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25030463

RESUMO

Breeding new varieties with low seed glucosinolate (GS) concentrations has long been a prime target in Brassica napus. In this study, a novel association mapping methodology termed 'associative transcriptomics' (AT) was applied to a panel of 101 B. napus lines to define genetic regions and also candidate genes controlling total seed GS contents. Over 100,000 informative single-nucleotide polymorphisms (SNPs) and gene expression markers (GEMs) were developed for AT analysis, which led to the identification of 10 SNP and 7 GEM association peaks. Within these peaks, 26 genes were inferred to be involved in GS biosynthesis. A weighted gene co-expression network analysis provided additional 40 candidate genes. The transcript abundance in leaves of two candidate genes, BnaA.GTR2a located on chromosome A2 and BnaC.HAG3b on C9, was correlated with seed GS content, explaining 18.8 and 16.8% of phenotypic variation, respectively. Resequencing of genomic regions revealed six new SNPs in BnaA.GTR2a and four insertions or deletions in BnaC.HAG3b. These deletion polymorphisms were then successfully converted into polymerase chain reaction-based diagnostic markers that can, due to high linkage disequilibrium observed in these regions of the genome, be used for marker-assisted breeding for low seed GS lines.


Assuntos
Brassica napus , Cromossomos de Plantas/fisiologia , Regulação da Expressão Gênica de Plantas/fisiologia , Glucosinolatos , Polimorfismo de Nucleotídeo Único , Sementes , Brassica napus/genética , Brassica napus/metabolismo , Mapeamento Cromossômico , Perfilação da Expressão Gênica , Glucosinolatos/biossíntese , Glucosinolatos/genética , Desequilíbrio de Ligação/fisiologia , Proteínas de Transporte de Monossacarídeos/biossíntese , Proteínas de Transporte de Monossacarídeos/genética , Sementes/genética , Sementes/metabolismo
8.
J Immunol ; 192(12): 5739-48, 2014 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-24835394

RESUMO

Cytokine-inducible SH2 domain-containing protein (CISH), a member of the suppressor of cytokine signaling family of negative feedback regulators, is induced by cytokines that activate STAT5 and can inhibit STAT5 signaling in vitro. However, demonstration of a definitive in vivo role for CISH during development has remained elusive. This study employed expression analysis and morpholino-mediated knockdown in zebrafish in concert with bioinformatics and biochemical approaches to investigate CISH function. Two zebrafish CISH paralogs were identified, cish.a and cish.b, with high overall conservation (43-46% identity) with their mammalian counterparts. The cish.a gene was maternally derived, with transcripts present throughout embryogenesis, and increasing at 4-5 d after fertilization, whereas cish.b expression commenced at 8 h after fertilization. Expression of cish.a was regulated by the JAK2/STAT5 pathway via conserved tetrameric STAT5 binding sites (TTCN3GAA) in its promoter. Injection of morpholinos targeting cish.a, but not cish.b or control morpholinos, resulted in enhanced embryonic erythropoiesis, myelopoiesis, and lymphopoiesis, including a 2- 3-fold increase in erythrocytic markers. This occurred concomitantly with increased activation of STAT5. This study indicates that CISH functions as a conserved in vivo target and regulator of STAT5 in the control of embryonic hematopoiesis.


Assuntos
Embrião não Mamífero/imunologia , Hematopoese/imunologia , Fator de Transcrição STAT5/imunologia , Proteínas Supressoras da Sinalização de Citocina/imunologia , Proteínas de Peixe-Zebra/imunologia , Peixe-Zebra/imunologia , Animais , Sequência de Bases , Hematopoese/genética , Janus Quinase 2/genética , Janus Quinase 2/imunologia , Dados de Sequência Molecular , Fator de Transcrição STAT5/genética , Proteínas Supressoras da Sinalização de Citocina/genética , Peixe-Zebra/genética , Proteínas de Peixe-Zebra/genética
9.
Clin Teach ; 10(6): 384-8, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24219523

RESUMO

BACKGROUND: Rural and island health care staff in Scotland are required to manage patients experiencing mental health crises. To ensure practitioners in remote and rural areas have the necessary skills, the Remote and Rural Healthcare Educational Alliance (RRHEAL) were asked to develop a pre-hospital mental health care course. METHODS: Several mental health care experts were asked to express an opinion on the essential content of such a course. Stakeholder review informed the development of a survey to identify the priority areas for training. The first round of the survey process involved an expert group of 16; the second round used a survey of over 300 remote and rural practitioners involved with the British Association for Immediate Care, Scotland (BASICS). RESULTS: The stakeholder review identified key content, summarised under the following topics: risk assessment; patient assessment; crisis management; handling difficult situations; engagement skills; mental health law; management of retrieval; pharmacology; theory and classification of mental illness; and understanding your network. DISCUSSION: This article shares how the needs within a national pre-hospital mental health care programme were identified, and demonstrates how consensus over the content of a national course was achieved through the use of a modified Delphi approach. The Clinical Skills Managed Educational Network (CSMEN), Self-Harm Mitigation Training (STORM®) and BASICS, alongside RRHEAL, contributed to this development. Using blended learning techniques the resulting course encourages a contextual approach to user needs, specifically teaching generic mental health care skills to staff who are often working single-handed, and are required to respond appropriately, locally and safely as they work towards achieving National Mental Health crisis standards.


Assuntos
Saúde Mental/educação , Saúde da População Rural/educação , Currículo , Técnica Delphi , Humanos , Transtornos Mentais/diagnóstico , Transtornos Mentais/terapia , Avaliação das Necessidades , Medição de Risco , Escócia
10.
J Biol Chem ; 288(52): 37267-76, 2013 Dec 27.
Artigo em Inglês | MEDLINE | ID: mdl-24220035

RESUMO

The proteoglycanase clade of the ADAMTS superfamily shows preferred proteolytic activity toward the hyalectan/lectican proteoglycans as follows: aggrecan, brevican, neurocan, and versican. ADAMTS15, a member of this clade, was recently identified as a putative tumor suppressor gene in colorectal and breast cancer. However, its biosynthesis, substrate specificity, and tissue expression are poorly described. Therefore, we undertook a detailed study of this proteinase and its expression. We report propeptide processing of the ADAMTS15 zymogen by furin activity, identifying RAKR(212)↓ as a major furin cleavage site within the prodomain. ADAMTS15 was localized on the cell surface, activated extracellularly, and required propeptide processing before cleaving V1 versican at position (441)E↓A(442). In the mouse embryo, Adamts15 was expressed in the developing heart at E10.5 and E11.5 days post-coitum and in the musculoskeletal system from E13.5 to E15.5 days post-coitum, where it was co-localized with hyaluronan. Adamts15 was also highly expressed in several structures within the adult mouse colon. Our findings show overlapping sites of Adamts15 expression with other members of ADAMTS proteoglycanases during embryonic development, suggesting possible cooperative roles during embryogenesis, consistent with other ADAMTS proteoglycanase combinatorial knock-out mouse models. Collectively, these data suggest a role for ADAMTS15 in a wide range of biological processes that are potentially mediated through the processing of versican.


Assuntos
Proteínas ADAM/biossíntese , Embrião de Mamíferos/metabolismo , Desenvolvimento Embrionário/fisiologia , Precursores Enzimáticos/biossíntese , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Regulação Enzimológica da Expressão Gênica/fisiologia , Versicanas/metabolismo , Proteínas ADAM/genética , Animais , Células COS , Chlorocebus aethiops , Precursores Enzimáticos/genética , Furina/genética , Furina/metabolismo , Células HEK293 , Humanos , Camundongos Knockout , Especificidade de Órgãos/fisiologia , Coelhos , Versicanas/genética
11.
BMC Plant Biol ; 13: 111, 2013 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-23915099

RESUMO

BACKGROUND: The detection and exploitation of genetic variation underpins crop improvement. However, the polyploid nature of the genomes of many of our most important crops represents a barrier, particularly for the analysis of variation within genes. To overcome this, we aimed to develop methodologies based on amplicon sequencing that involve the incorporation of barcoded amplification tags (BATs) into PCR products. RESULTS: A protocol was developed to tag PCR products with 5' 6-base oligonucleotide barcode extensions before pooling for sequencing library production using standard Illumina adapters. A computational method was developed for the de-convolution of products and the robust detection and scoring of sequence variants. Using this methodology, amplicons targeted to gene sequences were screened across a B. napus mapping population and the resulting allele scoring strings for 24 markers linkage mapped to the expected regions of the genome. Furthermore, using one-dimensional 8-fold pooling, 4608 lines of a B. napus mutation population were screened for induced mutations in a locus-specific amplicon (an orthologue of GL2.b) and mixed product of three co-amplified loci (orthologues of FAD2), identifying 10 and 41 mutants respectively. CONCLUSIONS: The utilisation of barcode tags to de-convolute pooled PCR products in multiplexed, variation screening via Illumina sequencing provides a cost effective method for SNP genotyping and mutation detection and, potentially, markers for causative changes, even in polyploid species. Combining this approach with existing Illumina multiplexing workflows allows the analysis of thousands of lines cheaply and efficiently in a single sequencing run with minimal library production costs.


Assuntos
Brassica napus/genética , Variação Genética , Poliploidia , Análise de Sequência de DNA/métodos , Produtos Agrícolas/genética , Ligação Genética , Genoma de Planta , Genótipo , Mutação , Proteínas de Plantas/genética , Polimorfismo de Nucleotídeo Único
12.
Mol Microbiol ; 89(6): 1167-86, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23869529

RESUMO

Plasmodium parasites remodel their vertebrate host cells by translocating hundreds of proteins across an encasing membrane into the host cell cytosol via a putative export machinery termed PTEX. Previously PTEX150, HSP101 and EXP2 have been shown to be bona fide members of PTEX. Here we validate that PTEX88 and TRX2 are also genuine members of PTEX and provide evidence that expression of PTEX components are also expressed in early gametocytes, mosquito and liver stages, consistent with observations that protein export is not restricted to asexual stages. Although amenable to genetic tagging, HSP101, PTEX150, EXP2 and PTEX88 could not be genetically deleted in Plasmodium berghei, in keeping with the obligatory role this complex is postulated to have in maintaining normal blood-stage growth. In contrast, the putative thioredoxin-like protein TRX2 could be deleted, with knockout parasites displaying reduced grow-rates, both in vivo and in vitro, and reduced capacity to cause severe disease in a cerebral malaria model. Thus, while not essential for parasite survival, TRX2 may help to optimize PTEX activity. Importantly, the generation of TRX2 knockout parasites that display altered phenotypes provides a much-needed tool to dissect PTEX function.


Assuntos
Parasitemia/parasitologia , Plasmodium berghei/enzimologia , Plasmodium berghei/patogenicidade , Tiorredoxinas/metabolismo , Fatores de Virulência/metabolismo , Animais , Modelos Animais de Doenças , Deleção de Genes , Malária Cerebral/parasitologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Plasmodium berghei/genética , Plasmodium berghei/crescimento & desenvolvimento , Análise de Sobrevida , Tiorredoxinas/genética , Virulência , Fatores de Virulência/genética
13.
Dev Biol ; 377(1): 46-54, 2013 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-23499657

RESUMO

Members of the Ikaros family of zinc-finger transcription factors have been shown to be critical for immune and blood cell development. However, the role of the most divergent family member, Pegasus, has remained elusive, although it shows conservation to invertebrate Hunchback proteins that influence embryonic patterning through regulation of homeodomain genes. Zebrafish was employed as a relevant model to investigate the function of Pegasus since it possesses a single pegasus orthologue with high homology to its mammalian counterparts. During zebrafish embryogenesis pegasus transcripts were initially maternally-derived and later replaced by zygotic expression in the diencephalon, tectum, hindbrain, thymus, eye, and ultimately the exocrine pancreas and intestine. Morpholino-mediated knockdown of the zebrafish pegasus gene resulted in disrupted left-right asymmetry of the gut and pancreas. Molecular analysis indicated that zebrafish Pegasus localised to the nucleus in discrete non-nucleolar structures and bound the 'atypical' DNA sequence GN3GN2G, confirming its presumed role as a transcriptional regulator. In vivo transcriptome analysis identified candidate target genes, several of which encoded homeodomain transcription factors. One of these, pitx2, implicated in left-right asymmetry, possessed appropriate 'atypical' Pegasus binding sites in its promoter. Knockdown of Pegasus affected both the level and asymmetry of pitx2 expression, as well as disrupting the asymmetry of the lefty2 and spaw genes, explaining the perturbed left-right patterning in pegasus morphants. Collectively these results provide the first definitive insights into the in vivo role of Pegasus, supporting the notion that it acts as a broader regulator of development, with potential parallels to the related invertebrate Hunchback proteins.


Assuntos
Padronização Corporal/genética , Regulação da Expressão Gênica no Desenvolvimento , Fator de Transcrição Ikaros/metabolismo , Fatores de Transcrição/genética , Proteínas de Peixe-Zebra/metabolismo , Peixe-Zebra/embriologia , Peixe-Zebra/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Padronização Corporal/efeitos dos fármacos , Embrião não Mamífero/efeitos dos fármacos , Embrião não Mamífero/metabolismo , Desenvolvimento Embrionário/efeitos dos fármacos , Desenvolvimento Embrionário/genética , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Técnicas de Silenciamento de Genes , Marcação de Genes , Células HEK293 , Humanos , Fator de Transcrição Ikaros/química , Fator de Transcrição Ikaros/genética , Dados de Sequência Molecular , Morfolinos/farmacologia , Fatores de Transcrição/metabolismo , Proteínas de Peixe-Zebra/química , Proteínas de Peixe-Zebra/genética
14.
J Biol Chem ; 288(3): 1907-17, 2013 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-23233679

RESUMO

Skeletal muscle development and regeneration requires the fusion of myoblasts into multinucleated myotubes. Because the enzymatic proteolysis of a hyaluronan and versican-rich matrix by ADAMTS versicanases is required for developmental morphogenesis, we hypothesized that the clearance of versican may facilitate the fusion of myoblasts during myogenesis. Here, we used transgenic mice and an in vitro model of myoblast fusion, C2C12 cells, to determine a potential role for ADAMTS versicanases. Versican processing was observed during in vivo myogenesis at the time when myoblasts were fusing to form multinucleated myotubes. Relevant ADAMTS genes, chief among them Adamts5 and Adamts15, were expressed both in developing embryonic muscle and differentiating C2C12 cells. Reducing the levels of Adamts5 mRNA in vitro impaired myoblast fusion, which could be rescued with catalytically active but not the inactive forms of ADAMTS5 or ADAMTS15. The addition of inactive ADAMTS5, ADAMTS15, or full-length V1 versican effectively impaired myoblast fusion. Finally, the expansion of a hyaluronan and versican-rich matrix was observed upon reducing the levels of Adamts5 mRNA in myoblasts. These data indicate that these ADAMTS proteinases contribute to the formation of multinucleated myotubes such as is necessary for both skeletal muscle development and during regeneration, by remodeling a versican-rich pericellular matrix of myoblasts. Our study identifies a possible pathway to target for the improvement of myogenesis in a plethora of diseases including cancer cachexia, sarcopenia, and muscular dystrophy.


Assuntos
Proteínas ADAM/metabolismo , Fibras Musculares Esqueléticas/metabolismo , Mioblastos/metabolismo , Regeneração , Versicanas/metabolismo , Proteínas ADAM/genética , Proteínas ADAMTS , Proteína ADAMTS5 , Animais , Comunicação Celular , Diferenciação Celular , Fusão Celular , Células Cultivadas , Embrião de Mamíferos , Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Células HEK293 , Humanos , Camundongos , Camundongos Transgênicos , Desenvolvimento Muscular , Fibras Musculares Esqueléticas/citologia , Fibras Musculares Esqueléticas/ultraestrutura , Mioblastos/citologia , Mioblastos/ultraestrutura , RNA Mensageiro/biossíntese , Trombospondinas/química
16.
BMC Genomics ; 13: 247, 2012 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-22703051

RESUMO

BACKGROUND: Polyploidy often results in considerable changes in gene expression, both immediately and over evolutionary time. New phenotypes often arise with polyploid formation and may contribute to the fitness of polyploids in nature or their selection for use in agriculture. Oilseed rape (Brassica napus) is widely used to study the process of polyploidy both in artificially resynthesised and natural forms. mRNA-Seq, a recently developed approach to transcriptome profiling using deep-sequencing technologies is an alternative to microarrays for the study of gene expression in a polyploid. RESULTS: Illumina mRNA-Seq is comparable to microarray analysis for transcript quantification but has increased sensitivity and, very importantly, the potential to distinguish between homoeologous genes in polyploids. Using a novel curing process, we adapted a reference sequence that was a consensus derived from ESTs from both Brassica A and C genomes to one containing separate A and C genome versions for each of the 94,558 original unigenes. We aligned reads from B. napus to this cured reference, finding 38% more reads mapping from resynthesised lines and 28% more reads mapping from natural lines. Where the A and C versions differed at single nucleotide positions, termed inter-homoeologue polymorphisms (IHPs), we were able to apportion expression in the polyploid between the A and C genome homoeologues. 43,761 unigenes contained at least one IHP, with a mean frequency of 10.5 per kb unigene sequence. 6,350 of the unigenes with IHPs were differentially expressed between homoeologous gene pairs in resynthesised B. napus. 3,212 unigenes showed a similar pattern of differential expression across a range of natural B. napus crop varieties and, of these, 995 were in common with resynthesised B. napus. Functional classification showed over-representation in gene ontology categories not associated with dosage-sensitivity. CONCLUSION: mRNA-Seq is the method of choice for measuring transcript abundance in polyploids due to its ability to measure the contributions of homoeologues to gene expression. The identification of large numbers of differentially expressed genes in both a newly resynthesised polyploid and natural B. napus confirms that there are both immediate and long-term alterations in the expression of homoeologous gene pairs following polyploidy.


Assuntos
Brassica napus/genética , Produtos Agrícolas/genética , Genoma de Planta/genética , Poliploidia , Análise de Sequência de RNA/métodos , Transcriptoma/genética , Sequência de Bases , Regulação da Expressão Gênica de Plantas , Genes de Plantas/genética , Dados de Sequência Molecular , Análise de Sequência com Séries de Oligonucleotídeos , Polimorfismo Genético , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Padrões de Referência , Homologia de Sequência do Ácido Nucleico , Especificidade da Espécie
17.
Nat Genet ; 43(10): 1035-9, 2011 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-21873998

RESUMO

We report the annotation and analysis of the draft genome sequence of Brassica rapa accession Chiifu-401-42, a Chinese cabbage. We modeled 41,174 protein coding genes in the B. rapa genome, which has undergone genome triplication. We used Arabidopsis thaliana as an outgroup for investigating the consequences of genome triplication, such as structural and functional evolution. The extent of gene loss (fractionation) among triplicated genome segments varies, with one of the three copies consistently retaining a disproportionately large fraction of the genes expected to have been present in its ancestor. Variation in the number of members of gene families present in the genome may contribute to the remarkable morphological plasticity of Brassica species. The B. rapa genome sequence provides an important resource for studying the evolution of polyploid genomes and underpins the genetic improvement of Brassica oil and vegetable crops.


Assuntos
Brassica rapa/genética , Genoma de Planta , Poliploidia , Arabidopsis/genética , Cromossomos Artificiais Bacterianos/genética , Cromossomos de Plantas/genética , Mapeamento de Sequências Contíguas , Evolução Molecular , Duplicação Gênica , Genes de Plantas , Anotação de Sequência Molecular , Dados de Sequência Molecular , Filogenia , Análise de Sequência de DNA
18.
Nat Biotechnol ; 29(8): 762-6, 2011 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-21804563

RESUMO

Polyploidy complicates genomics-based breeding of many crops, including wheat, potato, cotton, oat and sugarcane. To address this challenge, we sequenced leaf transcriptomes across a mapping population of the polyploid crop oilseed rape (Brassica napus) and representative ancestors of the parents of the population. Analysis of sequence variation and transcript abundance enabled us to construct twin single nucleotide polymorphism linkage maps of B. napus, comprising 23,037 markers. We used these to align the B. napus genome with that of a related species, Arabidopsis thaliana, and to genome sequence assemblies of its progenitor species, Brassica rapa and Brassica oleracea. We also developed methods to detect genome rearrangements and track inheritance of genomic segments, including the outcome of an interspecific cross. By revealing the genetic consequences of breeding, cost-effective, high-resolution dissection of crop genomes by transcriptome sequencing will increase the efficiency of predictive breeding even in the absence of a complete genome sequence.


Assuntos
Brassica rapa/genética , Genoma de Planta , Genômica/métodos , Análise de Sequência de DNA/métodos , Transcriptoma , Arabidopsis/genética , Brassica napus/genética , Mapeamento Cromossômico , Genes de Plantas , Polimorfismo Genético , Alinhamento de Sequência
19.
Genome Biol ; 11(9): R94, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20875114

RESUMO

BACKGROUND: The species Brassica rapa includes important vegetable and oil crops. It also serves as an excellent model system to study polyploidy-related genome evolution because of its paleohexaploid ancestry and its close evolutionary relationships with Arabidopsis thaliana and other Brassica species with larger genomes. Therefore, its genome sequence will be used to accelerate both basic research on genome evolution and applied research across the cultivated Brassica species. RESULTS: We have determined and analyzed the sequence of B. rapa chromosome A3. We obtained 31.9 Mb of sequences, organized into nine contigs, which incorporated 348 overlapping BAC clones. Annotation revealed 7,058 protein-coding genes, with an average gene density of 4.6 kb per gene. Analysis of chromosome collinearity with the A. thaliana genome identified conserved synteny blocks encompassing the whole of the B. rapa chromosome A3 and sections of four A. thaliana chromosomes. The frequency of tandem duplication of genes differed between the conserved genome segments in B. rapa and A. thaliana, indicating differential rates of occurrence/retention of such duplicate copies of genes. Analysis of 'ancestral karyotype' genome building blocks enabled the development of a hypothetical model for the derivation of the B. rapa chromosome A3. CONCLUSIONS: We report the near-complete chromosome sequence from a dicotyledonous crop species. This provides an example of the complexity of genome evolution following polyploidy. The high degree of contiguity afforded by the clone-by-clone approach provides a benchmark for the performance of whole genome shotgun approaches presently being applied in B. rapa and other species with complex genomes.


Assuntos
Brassica rapa/genética , Cromossomos de Plantas , Sequência Conservada , Análise de Sequência de DNA , Sintenia , Arabidopsis/genética , Sequência de Bases , Mapeamento Cromossômico , Estruturas Cromossômicas , Cromossomos Artificiais Bacterianos , Mapeamento de Sequências Contíguas , DNA de Plantas/genética , Evolução Molecular , Duplicação Gênica , Rearranjo Gênico , Genoma de Planta , Cariotipagem , Anotação de Sequência Molecular , Poliploidia
20.
Plant J ; 61(4): 591-9, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19929877

RESUMO

We conducted a sequence-level comparative analyses, at the scale of complete bacterial artificial chromosome (BAC) clones, between the genome of the most economically important Brassica species, Brassica napus (oilseed rape), and those of Brassica rapa, the genome of which is currently being sequenced, and Arabidopsis thaliana. We constructed a new B. napus BAC library and identified and sequenced clones that contain homoeologous regions of the genome including stearoyl-ACP desaturase-encoding genes. We sequenced the orthologous region of the genome of B. rapa and conducted comparative analyses between the Brassica sequences and those of the orthologous region of the genome of A. thaliana. The proportion of genes conserved (approximately 56%) is lower than has been reported previously between A. thaliana and Brassica (approximately 66%). The gene models for sets of conserved genes were used to determine the extent of nucleotide conservation of coding regions. This was found to be 84.2 +/- 3.9% and 85.8 +/- 3.7% between the B. napus A and C genomes, respectively, and that of A. thaliana, which is consistent with previous results for other Brassica species, and 97.5 +/- 3.1% between the B. napus A genome and B. rapa, and 93.1 +/- 4.9% between the B. napus C genome and B. rapa. The divergence of the B. napus genes from the A genome and the B. rapa genes was greater than anticipated and indicates that the A genome ancestor of the B. napus cultivar studied was relatively distantly related to the cultivar of B. rapa selected for genome sequencing.


Assuntos
Brassica napus/genética , Brassica rapa/genética , Hibridização Genômica Comparativa , Ácidos Graxos Dessaturases/genética , Genoma de Planta , Arabidopsis/enzimologia , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Brassica napus/enzimologia , Brassica rapa/enzimologia , Mapeamento Cromossômico , Cromossomos Artificiais Bacterianos , DNA de Plantas/genética , Biblioteca Gênica , Genes de Plantas , Modelos Genéticos , Filogenia , Análise de Sequência de DNA
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