Targeted gene modification in Fragaria vesca mediated by CRISPR/Cas9 system

Genome editing is becoming an important biotechnological tool for gene function analysis and crop improvement, being the CRISPR-Cas9 (Clustered Regularly Interspaced Short Palindromic Repeat-CRISPR associated protein 9) system the most widely used. The natural CRISPR/Cas9 system has been reduced to two components: a single-guide RNA (sgRNA) for target recognition via RNA-DNA base pairing, which is commonly expressed using a promoter for small-RNAs (U6 promoter), and the Cas9 endonuclease for DNA cleavage (1). To validate the CRISPR/Cas9 system in strawberry plants, we designed two sgRNAs directed against the floral homeotic gene APETALA3 (sgRNA-AP3#1 and sgRNA-AP3#2). This gene was selected because ap3 mutations induce clear developmental phenotypes in which petals and stamens are missing or partially converted to sepals and carpels respectively (2). In this work, we used two different U6 promoters to drive the sgRNA-AP3s expression: AtU6-26 from Arabidopsis (4), and a U6 promoter from Fragaria vesca (FvU6) (this work). We also tested two different coding sequences of Cas9: a human- (hSpCas9) (3) and a plant-codon optimized (pSpCas9) (this work). Transient expression experiments using both CRISPR/Cas9 systems (AtU6-26:sgRNA-AP3#1_35S:hSpCas9_AtU6-26:sgRNA-AP3#2 and FvU6:sgRNA-AP3#1_35S:pSpCas9_FvU6:sgRNA-AP3#2) were performed infiltrating Agrobacterium tumefaciens into F. vesca fruits. PCR amplification and sequencing analyses across the target sites showed a deletion of 188-189 bp corresponding to the region comprised between the two cutting sites of Cas9, confirming that the CRISPR/Cas9 system is functional in F. vesca. Remarkably, the two systems showed different mutagenic efficiency that could be related to differences in expression of the U6 promoters as well as differences in the Cas9 transcripts stability and translation. Stable transformants for both F. vesca (2n) and Fragaria X anannassa (8n) are currently being established to test whether is possible to obtain heritable homozygous mutants derived from CRISPR/Cas9 strategies in strawberry. Thus, our work offers a promising tool for genome editing and gene functional analysis in strawberry. This tool might represent a more efficient alternative to the sometimes inefficient RNAi silencing methods commonly used in this species.

Estimação e compensação de não linearidades inerentes aos atuadores dos processos industriais

The oscillations presents in control loops can cause damages in petrochemical industry. Canceling, or even preventing such oscillations, would save up to large amount of dollars. Studies have identified that one of the causes of these oscillations are the nonlinearities present on industrial process actuators. This study has the objective to develop a methodology for removal of the harmful effects of nonlinearities. Will be proposed an parameter estimation method to Hammerstein model, whose nonlinearity is represented by dead-zone or backlash. The estimated parameters will be used to construct inverse models of compensation. A simulated level system was used as a test platform. The valve that controls inflow has a nonlinearity. Results and describing function analysis show an improvement on system response

Qualidade da carne de ovinos puros e cruzados criados no Semiárido Nordestino brasileiro.

Resumo: A procura por carne de qualidade tem proporcionado ao setor de ovinocultura a adoção por técnicas de produção que possam contribuir para disponibilizar ao mercado produtos que atendam as necessidades dos consumidores, que se tornam cada vez mais exigentes quanto à aquisição de produtos que possam trazer benefícios a saúde. Assim, objetivou-se determinar a qualidade da carne de ovinos puros e cruzados criados no semiárido Nordestino. Utilizaram-se 30 cordeiros machos e fêmeas, oriundos dos cruzamentos Morada Nova x Morada Nova, Rabo Largo x Morada Nova e Santa Inês x Morada Nova, alojados em baias e alimentados com dieta a base de capim Canarana, milho farelo de soja e calcário. Os animais foram abatidos com peso médio de 24 kg de peso vivo e as carcaças mantidas em câmara frigorífica a 4?C, por 24 horas. Após esse período, foi seccionado o músculo Longissimus dorsi que foi embalado, identificado e armazenado a 20?C. Foram determinados os atributos físicos de capacidade de retenção de água, perda de peso por cocção e força de cisalhamento; os atributos químicos como umidade, proteína, cinza, lipídios e colesterol; avaliações subjetivas da carcaça como cor, marmoreio, textura da carne, distribuição da gordura, espessura da gordura e área de olho de lombo; os sensoriais de dureza, suculência, sabor, aroma e aceitação global e o perfil de ácidos graxos. O delineamento utilizado foi em esquema fatorial 3x2 (três grupos genéticos e dois sexos) e as médias comparadas pelo teste de Duncan a 5%. Não houve diferença (p>0,05) entre os genótipos para a composição química da carne, exceto para a variável lipídio (p<0,05). Para as características subjetivas da carcaça o genótipo influenciou (p<0,05) nas variáveis cor, marmoreio e textura da carne. O genótipo promoveu efeito (p<0,05) para as características físicas de capacidade de retenção de água e perda de peso por cocção. Os atributos sensoriais não sofreram efeitos (p>0,05) dos genótipos avaliados. Os genótipos influenciaram (p<0,05) as concentrações dos ácidos graxos saturados, monoinsaturados e poli-insaturados assim como, nas relações ?3:?6 e AGM:AGS. O fator sexo influenciou (p<0,05) nas variáveis umidade, força de cisalhamento, dureza, ácidos graxos desejáveis e relação ?3:?6. O cruzamento entre raças nativas mostrou-se com potencial para a produção de carne de qualidade, a raça Morada Nova melhorou os atributos físico-químicos da carne e em seu perfil lipídico e, o fator sexo melhorou o atributo maciez dando destaque a carne das fêmeas. Abstract: The demand for quality meat products has given the sheep industry sector productions techniques that can contribute providing the market with products that supply the needs of consumers require, who are becoming more demanding as the acquisition of differentiated and bring health benefits. The objective was to evaluate the quality of the pure sheep meat and cross bred in the semiarid Northeast of Brazil. It was used 30 male lambs and female, that came from crosses Morada Nova x Morada Nova, Rabo Largo x Morada Nova and Santa Ines x Morada Nova, housed in pens and fed a diet of grass Canarana, corn soybean meal and limestone. The animals were slaughtered at average weight of 24 kg, live weight, and carcasses kept in refrigeration chamber at 4 ° C for 24 hours. After this period the Longissimus dorsi muscle was selected, packaged, labeled and stored at 20˚C. The physical attributes were determined such as water holding capacity, cooking weight loss and shear force; the chemical attributes such as moisture, protein, ash, lipids and cholesterol; Subjective evaluations such as color, marbling, meat texture, fat distribution, fat thickness and loin eye area; the hardness sensory, juiciness, flavor, aroma and global acceptance and fatty acid profile. The design was a 3x2 factorial arrangement (three genetic groups and two genders) and the measures compared by Duncan test at 5%. There was no difference (p> 0.05) between genotypes for the chemical composition of meat, except for the lipid variable (p <0.05). To the subjective characteristics of the carcass, the genotype influenced (p <0.05) in the variables color, marbling and meat texture. The genotype promoted significant effect (p <0.05) for the physical characteristics, water retention capacity and weight loss on cooking. The sensory attributes did not suffer significant effects (p> 0.05) of the evaluated genotypes. The genotypes influenced (p <0.05) in the concentrations of saturated fatty acids, monounsaturated and polyunsaturated as well as in the relations ω3: ω6 and AGM:AGS. The gender factor influenced (P <0.05) in the variables moisture, shear strength, hardness, desirable fatty acids and relation ω3: ω6. The cross between native breeds showed up with potential for quality meat production, the Morada Nova improved the physical and chemical attributes of the flesh and in their lipid profiles and sex factor improved softness attribute highlighting the meat of females.

O óleo de patauá: seu estudo químico.

1951

Foreign manufacturing, regional agglomeration and technical efficiency in UK industries: a stochastic production frontier approach

The paper examines howfar foreign manufacturing investment in UK industries, together with the spatial agglomeration of those industries, affect technical efficiency. The paper links research on the estimation of technical efficiency,with those literatures demonstrating the economies associated with foreign direct investment and spatial agglomeration. The methodology involves estimation of a stochastic production frontier with random components associated with industry technical inefficiency, and a standard error. The paper also explores whether the degree of foreign involvement has a greater impact on technical efficiency where the domestic industry sector is characterized by comparatively high productivity and spatial agglomeration. The policy implications of the analysis are discussed.

MCMC Analysis for Optimization of Stochastic Models

In any decision making under uncertainties, the goal is mostly to minimize the expected cost. The minimization of cost under uncertainties is usually done by optimization. For simple models, the optimization can easily be done using deterministic methods.However, many models practically contain some complex and varying parameters that can not easily be taken into account using usual deterministic methods of optimization. Thus, it is very important to look for other methods that can be used to get insight into such models. MCMC method is one of the practical methods that can be used for optimization of stochastic models under uncertainty. This method is based on simulation that provides a general methodology which can be applied in nonlinear and non-Gaussian state models. MCMC method is very important for practical applications because it is a uni ed estimation procedure which simultaneously estimates both parameters and state variables. MCMC computes the distribution of the state variables and parameters of the given data measurements. MCMC method is faster in terms of computing time when compared to other optimization methods. This thesis discusses the use of Markov chain Monte Carlo (MCMC) methods for optimization of Stochastic models under uncertainties .The thesis begins with a short discussion about Bayesian Inference, MCMC and Stochastic optimization methods. Then an example is given of how MCMC can be applied for maximizing production at a minimum cost in a chemical reaction process. It is observed that this method performs better in optimizing the given cost function with a very high certainty.

Analysis of Some Stochastic Inventory Models with Pooling/ Retrial of Customers

In this thesis we have presented several inventory models of utility. Of these inventory with retrial of unsatisfied demands and inventory with postponed work are quite recently introduced concepts, the latt~~ being introduced for the first time. Inventory with service time is relatively new with a handful of research work reported. The di lficuity encoLlntered in inventory with service, unlike the queueing process, is that even the simplest case needs a 2-dimensional process for its description. Only in certain specific cases we can introduce generating function • to solve for the system state distribution. However numerical procedures can be developed for solving these problem.

Farm-level performance of genetically modified cotton - A frontier analysis of cotton production in Maharashtra

In this paper, the yield increases resulting from the cultivation of Bt cotton in Maharashtra, India, are analysed. The study relies on commercial farm, rather than trial, data and is among the first of its kind to be based on real farm and market conditions. Findings show that since its commercial release in 2002, Bt cotton has had a significant positive impact on yields and on the economic performance of cotton growers in Maharashtra. This difference remains even after controlling for different soil and insecticide inputs in the production of Bt cotton. There is also significant spatial and temporal variation in this 'benefit', and much depends upon where production is taking place and on the season.

Análise do desempenho muscular do quadríceps e dos isquiotibiais em função da série temporal e da amplitude de movimento de atletas amadoras de futsal feminino = : Analysis of muscle performance of the quadriceps and hamstrings as a function of the times series and range of motion of amateurfemale futsal athletes

Universidade Estadual de Campinas . Faculdade de Educação Física

Stochastic stability analysis for the constant-modulus algorithm

We derive an easy-to-compute approximate bound for the range of step-sizes for which the constant-modulus algorithm (CMA) will remain stable if initialized close to a minimum of the CM cost function. Our model highlights the influence, of the signal constellation used in the transmission system: for smaller variation in the modulus of the transmitted symbols, the algorithm will be more robust, and the steady-state misadjustment will be smaller. The theoretical results are validated through several simulations, for long and short filters and channels.

Analysis of glycosylation heterogeneity in antibody production by Pichia pastoris

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do grau de Mestre em Biotecnologia

Using time and stochastic models for software reliability forecasting and analysis

Tämä työ luo katsauksen ajallisiin ja stokastisiin ohjelmien luotettavuus malleihin sekä tutkii muutamia malleja käytännössä. Työn teoriaosuus sisältää ohjelmien luotettavuuden kuvauksessa ja arvioinnissa käytetyt keskeiset määritelmät ja metriikan sekä varsinaiset mallien kuvaukset. Työssä esitellään kaksi ohjelmien luotettavuusryhmää. Ensimmäinen ryhmä ovat riskiin perustuvat mallit. Toinen ryhmä käsittää virheiden ”kylvöön” ja merkitsevyyteen perustuvat mallit. Työn empiirinen osa sisältää kokeiden kuvaukset ja tulokset. Kokeet suoritettiin käyttämällä kolmea ensimmäiseen ryhmään kuuluvaa mallia: Jelinski-Moranda mallia, ensimmäistä geometrista mallia sekä yksinkertaista eksponenttimallia. Kokeiden tarkoituksena oli tutkia, kuinka syötetyn datan distribuutio vaikuttaa mallien toimivuuteen sekä kuinka herkkiä mallit ovat syötetyn datan määrän muutoksille. Jelinski-Moranda malli osoittautui herkimmäksi distribuutiolle konvergaatio-ongelmien vuoksi, ensimmäinen geometrinen malli herkimmäksi datan määrän muutoksille.

INCLUDING RISK IN ECONOMIC FEASIBILITY ANALYSIS:A STOCHASTIC SIMULATION MODEL FOR BLUEBERRY INVESTMENT DECISIONS IN CHILE

ABSTRACT The traditional method of net present value (NPV) to analyze the economic profitability of an investment (based on a deterministic approach) does not adequately represent the implicit risk associated with different but correlated input variables. Using a stochastic simulation approach for evaluating the profitability of blueberry (Vaccinium corymbosum L.) production in Chile, the objective of this study is to illustrate the complexity of including risk in economic feasibility analysis when the project is subject to several but correlated risks. The results of the simulation analysis suggest that the non-inclusion of the intratemporal correlation between input variables underestimate the risk associated with investment decisions. The methodological contribution of this study illustrates the complexity of the interrelationships between uncertain variables and their impact on the convenience of carrying out this type of business in Chile. The steps for the analysis of economic viability were: First, adjusted probability distributions for stochastic input variables (SIV) were simulated and validated. Second, the random values of SIV were used to calculate random values of variables such as production, revenues, costs, depreciation, taxes and net cash flows. Third, the complete stochastic model was simulated with 10,000 iterations using random values for SIV. This result gave information to estimate the probability distributions of the stochastic output variables (SOV) such as the net present value, internal rate of return, value at risk, average cost of production, contribution margin and return on capital. Fourth, the complete stochastic model simulation results were used to analyze alternative scenarios and provide the results to decision makers in the form of probabilities, probability distributions, and for the SOV probabilistic forecasts. The main conclusion shown that this project is a profitable alternative investment in fruit trees in Chile.

Asymptotic analysis of stock price densities and implied volatilities in mixed stochastic models

In this paper, we obtain sharp asymptotic formulas with error estimates for the Mellin con- volution of functions de ned on (0;1), and use these formulas to characterize the asymptotic behavior of marginal distribution densities of stock price processes in mixed stochastic models. Special examples of mixed models are jump-di usion models and stochastic volatility models with jumps. We apply our general results to the Heston model with double exponential jumps, and make a detailed analysis of the asymptotic behavior of the stock price density, the call option pricing function, and the implied volatility in this model. We also obtain similar results for the Heston model with jumps distributed according to the NIG law.