983 resultados para Clone


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A repetibilidade permite estimar o nmero de avaliaes ou ciclos produtivos para selecionar gentipos superiores com maior eficincia e menor custo operacional. Este trabalho estimou coeficientes de repetibilidade da produo, nmero e peso de fruto em 11 clones de cirigueleira, visando a avaliar a possibilidade de praticar seleo fenotpica individual. Foram analisados dados mdios de 5 safras, oriundos de 3 plantas por clone, propagadas por estaquia. Os coeficientes foram estimados pelos mtodos da anlise de varincia, componentes principais e anlise estrutural. O mtodo de componentes principais, baseados nas matrizes de varincias e covarincias, estima os maiores coeficientes em todas as caractersticas avaliadas. Com exceo do coeficiente estimado para nmero de frutos, pelo mtodo de componentes principais, todos os outros atingiram valores iguais ou menores que 0,38. De acordo com os resultados obtidos, possvel realizar seleo fenotpica individual entre os clones de cirigueleira avaliados, com base na produo e nmero de frutos por planta. As avaliaes de 6; 4 e 8 safras para produo, nmero e peso de frutos permitem selecionar clones promissores com cerca de 80 % de acurcia. O clone IPA-6 produziu 17,62 kg de frutos/planta, superando os demais materiais avaliados.

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Comparative analysis of gene fragments of six housekeeping loci, distributed around the two chromosomes of Vibrio cholerae, has been carried out for a collection of 29 V. cholerae O139 Bengal strains isolated from India during the first epidemic period (1992 to 1993). A toxigenic O1 ElTor strain from the seventh pandemic and an environmental non-O1/non-O139 strain were also included in this study. All loci studied were polymorphic, with a small number of polymorphic sites in the sequenced fragments. The genetic diversity determined for our O139 population is concordant with a previous multilocus enzyme electrophoresis study in which we analyzed the same V. cholerae O139 strains. In both studies we have found a higher genetic diversity than reported previously in other molecular studies. The results of the present work showed that O139 strains clustered in several lineages of the dendrogram generated from the matrix of allelic mismatches between the different genotypes, a finding which does not support the hypothesis previously reported that the O139 serogroup is a unique clone. The statistical analysis performed in the V. cholerae O139 isolates suggested a clonal population structure. Moreover, the application of the Sawyer's test and split decomposition to detect intragenic recombination in the sequenced gene fragments did not indicate the existence of recombination in our O139 population.

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RESUMO A podrido floral dos citros (PFC), causada por Colletotrichum acutatum Simmons e C. gloeosporioides, a doena fngica mais importante em limeira-cida ‘Tahiti’, pois, leva queda prematura de flores e frutos, acarretando a reduo da produo. Avaliou-se a suscetibilidade PFC dos clones de lima-cida ‘Tahiti’ “IAC 5”, “IAC 5-1”, “CNPMF/EECB”, “CNPMF 2000” e “CNPMF 2001”, em Bebedouro-SP. Todos os clones so suscetveis doena. A maior incidncia de PFC em plantas do clone “IAC 5-1” indica que este clone apresenta maior suscetibilidade. Mesmo com sintomas, plantas do clone “CNPMF/EECB” apresentam maior fixao de frutos.

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We investigated the decayed historical church window glasses of two Catalonian churches, both under Mediterranean climate. Glass surfaces were studied by scanning electron microscopy (SEM), energy dispersive spectrometry (EDS), and X-ray diffraction (XRD). Their chemical composition was determined by avelength-dispersive spectrometry (WDS) microprobe analysis. The biodiversity was investigated by molecular methods: DNA extraction from glass, amplification by PCR targeting the16S rRNA and ITS regions, and fingerprint analyses by denaturing gradient gel electrophoresis (DGGE). Clone libraries containing either PCR fragments of the bacterial 16S rDNA or the fungal ITS regions were screened by DGGE. Clone inserts were sequenced and compared with the EMBL database.

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In order to investigate a possible association between soybean malate synthase (MS; L-malate glyoxylate-lyase, CoA-acetylating, EC 4.1.3.2) and glyoxysomal malate dehydrogenase (gMDH; (S)-malate: NAD(+) oxidoreductase, EC 1.1.1.37), two consecutive enzymes in the glyoxylate cycle, their elution profiles were analyzed on Superdex 200 HR fast protein liquid chromatography columns equilibrated in low- and high-ionic-strength buffers. Starting with soluble proteins extracted from the cotyledons of 5-d-old soybean seedlings and a 45% ammonium sulfate precipitation, MS and gMDH coeluted on Superdex 200 HR (low-ionic-strength buffer) as a complex with an approximate relative molecular mass (M(r)) of 670000. Dissociation was achieved in the presence of 50 mM KCl and 5 mM MgCl2, with the elution of MS as an octamer of M, 510 000 and of gMDH as a dimer of M, 73 000. Polyclonal antibodies raised to the native copurified enzymes recognized both denatured MS and gMDH on immunoblots, and their native forms after gel filtration. When these antibodies were used to screen a lambda ZAP II expression library containing cDNA from 3-d-old soybean cotyledons, they identified seven clones encoding gMDH, whereas ten clones encoding MS were identified using an antibody to SDS-PAGE-purified MS. Of these cDNA clones a 1.8 kb clone for MS and a 1.3-kb clone for gMDH were fully sequenced. While 88% identity was found between mature soybean gMDH and watermelon gMDH, the N-terminal transit peptides showed only 37% identity. Despite this low identity, the soybean gMDH transit peptide conserves the consensus R(X(6))HL motif also found in plant and mammalian thiolases.

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Successful generation of high producing cell lines requires the generation of cell clones expressing the recombinant protein at high levels and the characterization of the clones' ability to maintain stable expression levels. The use of cis-acting epigenetic regulatory elements that improve this otherwise long and uncertain process has revolutionized recombinant protein production. Here we review and discuss new insights into the molecular mode of action of the matrix attachment regions (MARs) and ubiquitously-acting chromatin opening elements (UCOEs), i.e. cis-acting elements, and how these elements are being used to improve recombinant protein production. These elements can help maintain the chromatin environment of the transgene genomic integration locus in a transcriptionally favorable state, which increases the numbers of positive clones and the transgene expression levels. Moreover, the high producing clones tend to be more stable in long-term cultures even in the absence of selection pressure. Therefore, by increasing the probability of isolating a high producing clone, as well as by increasing transcription efficiency and stability, these elements can significantly reduce the time and cost required for producing large quantities of recombinant proteins.

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Infection with hepatitis E virus genotype 3 may result in chronic hepatitis in immunocompromised patients. Reduction of immunosuppression or treatment with ribavirin or pegylated interferon-α can result in viral clearance. However, safer and more effective treatment options are needed. Here, we show that sofosbuvir inhibits the replication of hepatitis E virus genotype 3 both in subgenomic replicon systems as well as a full-length infectious clone. Moreover, the combination of sofosbuvir and ribavirin results in an additive antiviral effect. Sofosbuvir may be considered as an add-on therapy to ribavirin for the treatment of chronic hepatitis E in immunocompromised patients.

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Nitric oxide (NO) has been shown to exert cytotoxic effects on tumor cells. We have reported that EC219 cells, a rat-brain-microvessel-derived endothelial cell line, produced NO through cytokine-inducible NO synthase (iNOS), the induction of which was significantly decreased by (a) soluble factor(s) secreted by DHD/PROb, an invasive sub-clone of a rat colon-carcinoma cell line. In this study, the DHD/PROb cell-derived NO-inhibitory factor was characterized. Northern-blot analysis demonstrated that the induction of iNOS mRNA in cytokine-activated EC219 cells was decreased by PROb-cell-conditioned medium. When DHD/PROb cell supernatant was fractionated by affinity chromatography using Con A-Sepharose or heparin-Sepharose, the NO-inhibitory activity was found only in Con A-unbound or heparin-unbound fractions, respectively, indicating that the PROb-derived inhibitory factor was likely to be a non-glycosylated and non-heparin-binding molecule. Pre-incubation of DHD/PROb-cell supernatant with anti-TGF-beta neutralizing antibody completely blocked the DHD/PROb-derived inhibition of NO production by EC219 cells. Addition of exogenous TGF-beta 1 dose-dependently inhibited NO release by EC219 cells. The presence of active TGF-beta in the DHD/PROb cell supernatant was demonstrated using a growth-inhibition assay. Moreover, heat treatment of medium conditioned by the less invasive DHD/REGb cells, which constitutively secreted very low levels of active TGF-beta, increased both TGF-beta activity and the ability to inhibit NO production in EC219 cells. Thus, DHD/PROb colon-carcinoma cells inhibited NO production in EC219 cells by secreting a factor identical or very similar to TGF-beta.

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Aim The reported prevalence of MET overexpression varies from 25-55% in non-small cell lung cancer (NSCLC) and clinical correlations are emerging slowly. In a well-defined NSCLC cohort of the Lungscape program, we explore the epidemiology, the natural history of IHC MET positivity and its association to OS, RFS and TTR. Methods Resected stage I-III NSCLC identified based on the quality of clinical data and FFPE tissue availability were assessed for MET expression using immunohistochemistry (IHC) on TMAs (CONFIRM anti total c-MET assay, clone SP44, Ventana BenchMark platform). All cases were analysed at participating pathology laboratories using the same protocol, after passing an external quality assurance program. MET positive status is defined as &#8805; 50% of tumor cells staining with 2+ or 3+ intensity. Results A total of 2709 cases are included in the iBiobank and will be analysed. IHC MET expression is currently available for 1552 patients, with positive MET IHC staining in 380 cases [24.5%; IHC 3+ in 157 cases (41.3%) and 2+ in 223 cases (58.7%)]. The cohort of 1552 patients includes 48.2%, 44.7% and 4.4% cases of adenocarcinoma, squamous and large cell histologies, respectively. IHC MET status was independent of stage, age and smoking history. Significant differences in MET positivity were associated with gender (32% vs. 21% for female vs. male, p < 0.001), with performance status (25% vs. 18% for 0 vs. 1-3, p = 0.006), and histology (34%, 14% and 24% for adenocarcinoma, squamous and large cell carcinoma, p < 0.001). IHC MET positivity was independent of the IHC ALK status (p = 0.08). At last FU, 52% of patients were still alive, with a median FU of 4.8 yrs. No association of IHC MET was found with OS, RFS or TTR. Conclusions The preliminary results for this large multicentre European cohort describe a prevalence of MET overexpression that seems lower than previous observations in NSCLC, such as reported for the OAM4971g trial, suggesting potential biological differences between surgically resected and metastatic disease. Analysis for the full cohort is ongoing and results will be presented. Disclosure L. Bubendorf: Disclosures: Stock ownership: Roche Advisory boards: Roche, Pfizer Research support: Roche; K. Schulze: Full time employee of Roche; A. Das-Gupta: I am a full time employee of Roche. All other authors have declared no conflicts of interest.

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AbstractObjective:In the present study, the authors investigated the in vitrobehavior of radio-resistant breast adenocarcinoma (MDA-MB-231) cells line and radiosensitive peripheral blood mononuclear cells (PBMC), as a function of different radiation doses, dose rates and postirradiation time kinetics, with a view to the interest of clinical radiotherapy.Materials and Methods:The cells were irradiated with Co-60, at 2 and 10 Gy and two different exposure rates, 339.56 cGy.min&#8211;1 and the other corresponding to one fourth of the standard dose rates, present over a 10-year period of cobalt therapy. Post-irradiation sampling was performed at pre-established kinetics of 24, 48 and 72 hours. The optical density response in viability assay was evaluated and a morphological analysis was performed.Results:Radiosensitive PBMC showed decrease in viability at 2 Gy, and a more significant decrease at 10 Gy for both dose rates. MDAMB- 231 cells presented viability decrease only at higher dose and dose rate. The results showed MDA-MB-231 clone expansion at low dose rate after 48&#8211;72 hours post-radiation.Conclusion:Low dose rate shows a possible potential clinical impact involving decrease in management of radio-resistant and radiosensitive tumor cell lines in cobalt therapy for breast cancer.

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UNLABELLED: Whole-genome sequencing (WGS) of 228 isolates was used to elucidate the origin and dynamics of a long-term outbreak of methicillin-resistant Staphylococcus aureus (MRSA) sequence type 228 (ST228) SCCmec I that involved 1,600 patients in a tertiary care hospital between 2008 and 2012. Combining of the sequence data with detailed metadata on patient admission and movement confirmed that the outbreak was due to the transmission of a single clonal variant of ST228, rather than repeated introductions of this clone into the hospital. We note that this clone is significantly more frequently recovered from groin and rectal swabs than other clones (P &lt; 0.0001) and is also significantly more transmissible between roommates (P &lt; 0.01). Unrecognized MRSA carriers, together with movements of patients within the hospital, also seem to have played a major role. These atypical colonization and transmission dynamics can help explain how the outbreak was maintained over the long term. This "stealthy" asymptomatic colonization of the gut, combined with heightened transmissibility (potentially reflecting a role for environmental reservoirs), means the dynamics of this outbreak share some properties with enteric pathogens such as vancomycin-resistant enterococci or Clostridium difficile. IMPORTANCE: Using whole-genome sequencing, we showed that a large and prolonged outbreak of methicillin-resistant Staphylococcus aureus was due to the clonal spread of a specific strain with genetic elements adapted to the hospital environment. Unrecognized MRSA carriers, the movement of patients within the hospital, and the low detection with clinical specimens were also factors that played a role in this occurrence. The atypical colonization of the gut means the dynamics of this outbreak may share some properties with enteric pathogens.

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En aquest projecte crearem un sistema per automatitzar els diferents dispositius que podem trobar en una casa. En primer lloc dissenyarem el hardware que ser el sistema nervis des del que controlarem els dispositius a travs del port USB dun ordinador. Aquest sistema nervis ser el punt dinterconnexi entre els dispositius de la casa i lordinador central que els controlar. A nivell de hardware, a ms a ms del mdul dentrades i sortides dinterconnexi amb els dispositius que hem esmentat, ens trobem amb la necessitat dinstallar un ordinador central i diferents aparells repartits per la casa per poder realitzar les nostres necessitats (accions dels diferents dispositius) des de qualsevol punt de la casa. Amb aquests requeriments haurem destudiar les diferents possibilitats per fer el nostre sistema el mxim defica possible. Finalitzat lestudi del hardware necessari pel nostre projecte, el segent pas s dissenyar el software. Aquest software ser laplicaci encarregada de controlar tot el maquinari que hem dissenyat anteriorment i rebr el nom de DOMO HOGAR. Aquest estar format per dos programes diferents, DOMO HOGAR SERVER i DOMO HOGAR TERMINAL, cadascun dells amb unes funcions especfiques. DOMO HOGAR SERVER ser laplicaci que residir a lordinador central i que permetr a ladministrador gestionar totes les parts de les que forma part el nostre sistema: dispositius, tasques, pre-condicions, etc... Tamb des daquesta aplicaci editarem el panell tctil que mostrarem des dels diferents terminals de lhabitatge. Per ltim, aquesta aplicaci tamb sencarregar de resoldre les peticions que farem, tant de lordinador central com dels terminals, i gestionar les diferents sortides en funci de lacci a realitzar. Parallelament ens trobarem laplicaci DOMO HOGAR TERMINAL que residir en cada un dels terminals que hi hagi a la casa. Aquesta aplicaci sinicialitzar llegint la configuraci del panell tctil de la base de dades de laplicaci servidor resident a lordinador central i reconstruint una rplica daquest panell tctil. Finalment des daquesta aplicaci terminal podrem donar ordres que seran emmagatzemades a la llista de tasques pendents de lordinador central perqu les resolgui des de laplicaci del servidor. DOMO HOGAR ha estat creat per facilitar i confortar la vida quotidiana de les persones agilitzant el nostre dia a dia i permetent-nos invertir el nostre temps en les coses realment importants.

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Seedlings of 41 different citrus species and varieties were massively colonized with the citrus brown aphid Toxoptera citricidus, obtained from Pra sweet orange (Citrus sinensis) trees, presenting symptoms of the "Capo Bonito" complex of the Citrus tristeza virus (CTV). The objective was to evaluate resistance or tolerance of the varieties to that virus complex, but even after eight months of inoculation no stem pitting was observed in the plants. Otherwise, the presence of galls similar to those induced by the vein enation-woody gall disease was observed in 73% of the plants of Volkamer Palermo (Citrus volkameriana), 60% of the Volkamer Catania 2, 2% of the Rangpur Lime D.22.30 (Citrus limonia), 13% of the Volkamer Australian Red, the Rangpur Lime hybrid, the Orlando tangelo (Citrus reticulata x Citrus paradisi) and the Florida Rough lemon (C. jambhiri), and 7% of the Carrizo citrange (Poncirus trifoliata x Citrus sinensis). The highest incidence and the largest gall size were observed in the Palermo Volkamer showing that this clone would be the most suitable to be used as an indicator plant in biological indexing tests for the disease. There are no previous reports in the literature about the occurrence of woody galls in Orlando tangelo and Carrizo citrange.

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Foi estudado o processo de infeco de Colletotrichum guaranicola em clones de guaranazeiro (Paullinia cupana var. sorbilis) resistente e suscetvel, por meio de microscopia de luz. Avaliaram-se quantitativamente os eventos de pr-penetrao em folhas novas e velhas dos clones. Observou-se diferena quantitativa quanto formao de apressrio em folhas novas e velhas dos clones, sendo maior em folhas novas do clone suscetvel. Estruturas de pr-penetrao e infeco foram semelhantes nos dois clones. O processo de infeco teve inicio aps a germinao do fungo e formao de apressrio na superfcie do hospedeiro, seguido de penetrao direta da cutcula e da parede celular das clulas epidrmicas. Dentro da clula, a hifa de infeco deu origem a uma vescula, que em seguida formou a hifa primria que se ramificou, originando a hifa secundria, que colonizou intra e intercelularmente a epiderme e o parnquima, causando desorganizao dos tecidos e necrose. Observou-se uma diferena temporal na colonizao dos tecidos dos clones. No clone suscetvel, com 48 h aps a inoculao, as clulas da epiderme e do parnquima estavam colonizadas por hifas intra e intercelulares. No quinto dia aps a inoculao, observou-se o surgimento dos sintomas. No clone resistente, a colonizao s foi observada no quarto dia aps a inoculao. Somente no stimo dia aps a inoculao, foram observados os primeiros sintomas tpicos da doena.

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The purpose of this research was to evaluate the differences in the colonization and production of structures in the leaves of 'Pra' sweet orange (Citrus sinensis) clones and related varieties by Guignardia citricarpa. The natural colonization and the production of reproductive structures in the leaves and in vitro of ten 'Pra' sweet orange was quantified in the following clones: Bianchi, Dibbern C.V., EEL, IAC 2000, Olmpia 15161, Premunizada 1212, Premunizada 1743/82, R. Gullo 1569/244, R. Gullo 1570/246 and Vimusa; and in five related varieties: Redonda C.N, Ovale 968, Ovale San Lio 969, Lamb Summer and Corsa Tardia. The quantification of the colonization density of G. citricarpa in the leaves was obtained through isolation. Incidence and colonization density (cm) were calculated for each clone. The production of reproductive structures was accomplished through the moistening and drying process of the leaves. The incidence (percentage of affected leaves) and the leaf surface percentage occupied by the reproductive fungus structures were quantified. The in vitro production of reproductive structures was accomplished in water-agar medium. The number of immature and total reproductive fungus structures (cm), and the percentage of picnidia with liberation of spores were quantified. Significant differences were not observed among clones related to the colonization of the leaves. But there were differences in the induction experiments, i.e., in the leaf surface percentage occupied by the reproductive fungus structures and the in vitro production of reprodutive fungus structures.