Enhancement of non-viral transfection efficiency with nuclear localization signal peptides

Dissertação de Mestrado, Ciências Biomédicas, Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve, 2016

Influencia de fontes e niveis de nitrogenio na cultura da cana-de-acucar irrigada.

Realizou-se um experimento em um Vertissolo do Submedio Sao Francisco, para avaliar a influencia de fontes e niveis de nitrogenio na produtiivdade da cana-de-acucar (Saccharum officinarum L.). O delineamento usado foi o de blocos ao acaso, com tres repeticoes e 14 tratamentos dispostos da seguinte maneira: 0, 40, 80, 120, 160 e 200 kg/ha de N sob a forma de ureia; 40, 80, 120, 160 e 200 kg/ha de N, sob a forma de sulfato de amonio; e 80, 120 e 160 kg/ha de N, sendo 50% sob a forma de ureia aplicada no plantio e 50% sob a forma de sulfato de amonio aplicado 120 dias apos o plantio. Os resultados mostraram que nao houve influencia das fontes de nitrogenio na produtividade da cana. Os niveis de nitrogenio exerceram uma influencia positiva e altamente significativa na produtividade. A dose economica de nitrogenio foi a de 213 kg/ha de N. Houve correlacoes lineares e positivas dos teores de nitrogenio na folha com os niveis de N aplicado e com a produtividade da cana. Nenhuma influencia foi verificada dos niveis de nitrogenio nos teores de acucar da cana.

Análise comparativa entre as propriedades do gesso obtido de rejeito da produção de sal e gesso comerciais

The environmental impacts, caused by the solid residues generation, are an often quoted concern nowadays. Some of these residues, which are originated from different human activities, can be fully reused, reducing the effects of the poor waste management on the environment. During the salt production process, the first formed crystals are discarded as industrial waste. This is mainly made of gypsum that is a calcium sulfate dihydrate (CaSO4.2H2O). The gypsum in question may go through a calcination process due to the plaster (CaSO4.0,5H2O) production and then the application on the cement industry. Considering the necessity of development and application for these industrial wastes, this paper aims to analyze the plaster, called Salgesso, from the gypsum that was generated during the salt production, and its use viability on the civil construction industry in order to create environmental and economical benefits. For characterization, the following experiments were performed: X-ray Fluorescence (XRF), X-ray Diffraction (XRD), thermal analysis (TG/DTG) and Scanning Electron Microscopy (SEM) with EDS. The following tests were also performed to obtain the mechanical characteristics: Thinness Modulus, Unit Mass, Setting Time and Compressive Resistance. Three commercial plasters used on civil construction were taken as references. All of these tests were performed according to the current standards. It was noticed that although there were some conflicting findings between the salt and commercial plasters in all of the studied properties, the Salgesso has its values within the standard limits. However, there is the possibility to improve them by doing a more effective calcination process. Three commercial plasters, used in construction, were used as reference material. All tests were performed according to standards in force. It was observed that although some tests present conflicting findings between the salt and gypsum plasters commercial properties in all of the studied Salgesso have values within the limits imposed by the standard, but can be improved simply by calcination process more effective

Desenvolvimento de argamassa para restauração utilizando resíduo do polimento do porcelanato

The coatings mortars are essential elements of building structures because they execute an important role in protecting walls and are particularly exposed to aggressive action responsible for its degradation over time. The importance of wall coverings has been the subject of discussion and analysis in the conservation and rehabilitation of old buildings. Are sometimes removed and replaced with inappropriate solutions of constructive point of view or architecture. The most commonly used coatings on walls of old buildings is based on traditional hydraulic lime mortars. The present study aims at the formulation of new lime- based mortars and aerial fine aggregate, in order to contribute to a better field of conservation and restoration mortar coating of old buildings. Residue was used for polishing porcelain as fine aggregate, replacing the aggregate (sand), in percentages 05-30% by mass. We conducted a thorough evaluation of the mortar properties in fresh and hardened state by comparing the performance of the same with a reference mortar. The residue used was characterized as the density, bulk density, and particle size laser, scanning electron microscopy, X-ray diffraction and X-ray fluorescence. Formulations were produced 7, 6 with residue and one commonly used formulation, which served as a reference. In the formulations of lime mortars air (hydrated lime powder CH-I) has been adopted a stroke volume (1:3) with constant binder, was varied and the water / binder and aggregate and waste. For evaluation of mortars fresh, proceeded to consistency analysis, specific gravity, water retention and air content embedded. In the hardened state assays were performed in specific gravity, water retention, modulus of elasticity, tensile strength in bending, compressive strength, water absorption by capillary action, adhesion, tensile strength, resistance to shrinkage and salts by of crystallization trials with resources chloride solution, nitrate and sulfate all sodium in prismatic at 90 days of age, in addition to the micro structural analysis of mortars. Based on the results we can see that the mortar formulated with 10% content of waste and the reference free retraction feature more stable closer to neutrality. The composition of 10% was obtained better performance against the action of the salt crystallization. The mortar with 15% residue obtained better density, lower air content embedded and high capacity for water retention developing good workability. The replacement of 20% of waste generates a satisfactory utilization of resistance to compression, flexion and traction grip the base. And, finally, it can be seen that the mortar with 10, 15 and 20% residual show, in principle, good suitability as coatings, thus enabling a final result consistent with durability, workability and aesthetics developing therefore a material with better performance to repair or replace existing mortars in old buildings

Teor de clorofila e produtividade do trigo em razão da adubação foliar com boro na presença de aminoácidos.

O trigo é uma cultura de ciclo anual cultivada no inverno. Nas culturas de arroz, trigo, aveia e milho, foram evidenciados boas correlações entre a leitura SPAD e a produtividade de grãos. O trabalho teve como objetivo estudar o efeito da adubação foliar com boro (B) na presença de aminoácidos (AAs) no teor de clorofila e produtividade de grãos do trigo, assim como na sua correlação. O experimento foi realizado na Embrapa Soja (Londrina - PR), no ano de 2014. O solo da área experimental é classificado como Latossolo Vermelho distroférrico. O delineamento experimental foi em blocos ao acaso, com quatro repetições e seis tratamentos com doses de ácido bórico na presença de aminoácidos, exceto a testemunha. As doses estudadas foram: T1- 0 kg ha-1 de H3BO3 sem aminoácidos (Testemunha); T2- 0 kg ha-1 de H3BO3; T3- 1 kg ha-1 de H3BO3; T4- 2 kg ha-1 de H3BO3; T5- 4 kg ha-1 de H3BO3; T6- 8 kg ha-1 de H3BO3. O plantio da cultivar BRS Pardela foi realizado com semeadora com sistema de plantio direto, com espaçamento entre linha de 50 cm e a densidade de semeadura utilizada foi ajustada para atingir 250.000 plantas ha-1. A adubação de semeadura foi realizada com 40 kg ha-1 de N (ureia), 60 kg ha-1 de P2O5 (superfosfato triplo) + 40 kg ha-1 K2O (KCl) + 2,0 kg ha-1 de Cu (sulfato de cobre 24,5%), 2,0 kg ha-1 de Mn (sulfato de manganês 30%) e 2,0 kg ha-1 de Zn (sulfato de zinco 21%). A adubação em cobertura foi realizada aos 40 dias após o plantio com 20 kg ha-1 K2O (KCl). A adubação foliar com boro e AAs (2 L ha-1) foi realizada na fase final de alongamento e início de pré-espigamento. Foi utilizado o ácido bórico (17%B) e os aminoácidos na concentração (1/200, v/v) com 6,8% de glicina, 4,4% de prolina, 3,3% de ácido glutâmico, 2,7% de alanina, 1,9% de arginina, 1,7% de ácido aspártico, 1,3% de lisina, 1,3% de histidina e 1,0% de leucina. As práticas de proteção de plantas via controle químico de pragas, doenças e plantas daninhas foram realizadas conforme as indicações técnicas para a cultura de trigo a fim de não interferirem nos resultados obtidos. Avaliou-se a produtividade de grãos de trigo e o teor de clorofila, determinado indiretamente no terço médio da folha bandeira de cinco plantas por parcela, no florescimento, por meio de leituras SPAD com auxílio de um clorofilômetro digital (Minolta 502). Os dados foram submetidos à análise de variância pelo teste F. A comparação de médias pelo teste de Tukey e as correlações foram feitas em nível de 5% de significância. Não houve efeito significativo dos tratamentos no teor de clorofila (SPAD) e na produtividade de grãos de trigo. A produtividade média de grãos foi de 4.266 kg ha-1 e 40 SPAD. Não houve correlação entre a leitura SPAD e a produtividade de grãos. Conclui-se que a adubação foliar com boro na presença de aminoácidos não modificou o teor de clorofila e a produtividade de grãos do trigo, bem como não houve correlação entre os fatores.

Carbono da biomassa microbiana em solo com aplicação de dejetos de suínos em sistema de plantio direto e convencional.

A aplicação de dejetos de suínos em solos cultivados pode alterar suas características químicas, físicas e biológicas. Estes estudos são importantes para compreensão e recomendação de práticas conservacionistas de uso do solo, que visam minimizar os efeitos da produção agrícola nos recursos naturais. O objetivo deste trabalho foi avaliar o carbono da biomassa microbiana (CBM) do solo, em três profundidades, após a aplicação de cinco doses de dejetos líquidos de suínos (DLS) em sistema de plantio direto (SPD) e convencional (SPC). O experimento foi instalado em 1997, na Estação Experimental do Instituto Agronômico do Paraná, em Palotina, Paraná, em Latossolo Vermelho distroférrico de textura argilosa. Os tratamentos foram a aplicação de 0, 30, 60, 90 e 120 m3 ha-1 ano-1 de dejetos de suínos no solo cultivado em SPD ou SPC, em delineamento experimental de blocos casualizados, com três repetições. Em outubro de 2015, após o cultivo de aveia (Avena strigosa), foram coletadas amostras de solo, nas profundidades de 0-10 cm, 10-20 cm e 20-40 cm, para determinação dos atributos químicos e do CBM. O DLS apresentou em g L-1 58,20 de massa seca; 5,52 N; 3,97 P; 1,4 K; 3,32 Ca; 2,16 Mg; 0,01 Cu; 0,23 Zn; 0,06 Mn; densidade de 1,01 g cm-3 e condutividade de 10,72 mS cm-1. O CBM foi avaliado pelo método de fumigação-extração. O teor de carbono nos extratos foi quantificado por titulação com sulfato ferroso amoniacal, após a oxidação do dicromato de potássio por digestão sulfúrica. O CBM foi calculado utilizando um fator de correção Kc de 0,33. A análise de variância foi realizada a 5% de probabilidade e, quando significativo pelo teste F, as médias foram comparadas pelo teste de Tukey. O CBM, com a aplicação de DLS no SPD, apresentou valores médios de 457, 355 e 284 micrograma C-CBM g solo nas profundidades de 0-10, 10-20 e 20-40 cm, respectivamente. No SPC, o CBM, com a aplicação de DLS, apresentou valores médios de 400, 284 e 253 micrograma C-CBM g solo, respectivamente, nas mesmas profundidades. Não houve efeito significativo das doses de DLS sobre o CBM nas profundidades 0-10, 10-20 e 20-40 cm, tanto sob SPD quanto SPC. Os valores de CBM foram maiores no SPD quando comparados com SPC na profundidade de 0-10 cm e 10-20 cm, não diferindo entre si na profundidade de 20-40 cm. Houve interação significativa entre as doses de DLS e SPD sobre o CBM, somente na adição das doses de 0 e 120 m3 ha-1 ano-1 DLS na profundidade de 10-20 cm.

Biological and chemical strategies for the recovery of precious and rare metals as nanoparticles

Tese de Doutoramento, Ciências do Mar da Terra e do Ambiente, Ramo: Ciências e Tecnologias do Ambiente, Especialidade em Biotecnologia, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2016

Estudo do resíduo fosfogesso na formulação de uma pasta para revestimento interno de edifícios

Universidade Federal do Rio Grande do Norte

Tecnologia de semeadura cruzada e reduzida na cultura da soja em função de doses de boro e zinco.

O objetivo do trabalho foi o de comparar a resposta de dois sistemas de semeadura nas características agronômicas da cultura da soja. O experimento foi conduzido na safra 2013/2014, no município de Sinop-MT, na Embrapa Agrossilvipastoril. Utilizou-se a cultivar BRS 8381, convencional. Realizou-se semeadura Cruzada a 0,45 m e semeadura Reduzida a 0,23 m entre linhas, mantendo-se a mesma população de plantas por hectare (POP). Os demais fatores experimentais foram constituídos por 3 doses de B (0, 1 e 2 kg ha-1) e 3 doses de Zn (0, 2 e 4 kg ha-1), utilizando-se como fontes o ácido bórico e o sulfato de zinco. Os dados foram submetidos à análise de variância e as médias comparadas pelo teste de Tukey, a 5% de probabilidade. Avaliou-se os seguintes parâmetros: POP, massa de 100 grãos (M100), número de vagens por planta (VAG), altura de plantas (ALT) e a produtividade de grãos (PROD). Para a POP, observou-se diferenças significativas entre os arranjos de planta (arranjos) e também na interação entre B e Zn. Entre os arranjos, o Cruzado foi estatisticamente superior ao Reduzido, com 410.000 pl ha-1 e 315.000 pl ha-1, respectivamente. Entre as doses de B dentro de Zn, observou-se diferenças somente para o tratamento testemunha de Zn, sendo que a menor dose de B proporcionou maior POP (396.000 pl ha-1) que a maior dose (343.000 pl ha-1), porém, semelhantes a dose intermediária (359.000 pl ha-1). Quanto a M100, observou-se diferenças somente entre os arranjos. O arranjo Reduzido (12,8g) proporcionou maior M100 em relação ao arranjo Cruzado (12,3 g). Para a ALT, observouse diferenças somente entre os arranjos avaliados. O Reduzido apresentou maior ALT, com 80 cm, em relação ao Cruzado (74 cm). Esse fato se deve ao próprio arranjo, onde as plantas competem mais por luz entre si, na fileira, em relação ao arranjo cruzado, causando aumento em altura. Em relação ao VAG, observou-se diferenças entre os arranjos e entre as doses de Zn. Entre os arranjos, o sistema Reduzido (45 vag pl-1) foi superior ao Cruzado (38 vag pl-1). Observou-se também redução do VAG com o aumento da POP, o que pode ser explicado pela plasticidade da cultura, que compensou o VAG pela diferença na POP. Em relação as doses de Zn, observou-se que a maior dose proporcionou maior VAG (44 vag pl-1) em relação a testemunha (39 vag pl-1), porém, semelhantes a dose intermediária (41 vag pl-1). Em relação à PROD, somente os rranjos se diferenciaram entre si. A maior PROD foi obtida no arranjo Reduzido (2738 kg ha-1) em relação ao Cruzado (2253 kg ha-1). Conclui-se que nas condições em que se desenvolveu o trabalho, o arranjo Reduzido foi superior. A adubação com B, se mal manejada, pode reduzir a POP e a aplicação adequada de Zn pode proporcionar aumento do VAG.

Avaliação de produtos naturais no controle de biofilmes formados em sistema dinâmico

The biofilms microbial forms of association are responsible for generating, accelerating and / or induce the process of corrosion. The damage generated in the petroleum industry for this type of corrosion is significatives, representing major investment for your control. The aim of this study was to evaluate such tests antibiograms the effects of extracts of Jatropha curcas and essential oil of Lippia gracilis Schauer on microrganisms isolated from water samples and, thereafter, select the most effective natural product for further evaluation of biofilms formed in dynamic system. Extracts of J. curcas were not efficient on the complete inhibition of microbial growth in tests type antibiogram, and essential oil of L. gracilis Schauer most effective and determined for the other tests. A standard concentration of essential oil of 20 μL was chosen and established for the evaluation of the biofilms and the rate of corrosion. The biocide effect was determined by microbial counts of five types of microorganisms: aerobic bacteria, precipitating iron, total anaerobic, sulphate reducers (BRS) and fungi. The rate of corrosion was measured by loss of mass. Molecular identification and scanning electron microscopy (SEM) were performed. The data showed reduction to zero of the most probable number (MPN) of bacteria precipitating iron and BRS from 115 and 113 minutes of contact, respectively. There was also inhibited in fungi, reducing to zero the rate of colony-forming units (CFU) from 74 minutes of exposure. However, for aerobic and anaerobic bacteria there was no significant difference in the time of exposure to the essential oil, remaining constant. The rate of corrosion was also influenced by the presence of oil. The essential oil of L. gracilis was shown to be potentially effective

Avaliação da biodessulfurização de 4-metildibenzotiofeno por Rhodococus rhodochrous (NRRL B-2149)

Crude oil has between 0.04 up to 5% of sulphur, the higher the oil the higher the sulphur levels. Sulphur usually gives problems such as corrosion in refinery, and once burnt produces SO2 that goes to atmosphere. This work aim to investigate the capacity of Rhodococcus rhodochrous (NRRL B-2149) to metabolize the model compound 4-methyldibenzotiophene (4-MDBT), to remove the sulphur and transform it in 2-hydroxybiphenyl (2-HBF) and sulphite using the 4S pathway. Kynetic runs were carried out in shaker at 120 rpm and 32°C. Samples were taken every 12h to assay substrate consume as well as cells production using HPLC. Results showed that R. rhodochrous NRRL B-2149 can use the 4S pathway in order to remove sulphur without change the carbon chain of the molecule as well as that cells and 4-MDBT affects the product formation. The production of 2-hydroxybiphenyl has interest for industry once it is a potent biocide. However, evaluation is necessary in order to obtain better results compatible with industry needs

Calda bordalesa: utilidades e preparo.

2016

Extração de metais (K, Mg, Ba, Ca, Cr, Mn, Li, Fe) de água produzida utilizando sistema microemulsionado

The produced water is a byproduct formed due to production of petroleum and carries with it a high amount of contaminants such as oil particles in suspension, organic compounds and metals. Thus, these latter pollutants are very difficult to treat because of its high solubility in water. The objective of this work is to use and evaluate a microemulsioned system to remove metals ( K , Mg , Ba , Ca , Cr , Mn , Li , Fe ) of synthetic produced water. For the extraction of metals, it was used a pseudoternary diagram containing the following phases: synthetic produced water as the aqueous phase (AP), hexane as organic phase (OP), and a cosurfactant/surfactant ratio equal to four (C/S = 4) as the third phase, where the OCS (saponified coconut oil) was used as surfactant and n-butanol as cosurfactant. The synthetic produced water was prepared in a bench scale and the region of interest in the diagram for the removal of metals was determined by experimental design called. Ten points located in the phase Winsor II were selected in an area with a large amount of water and small amounts of reagents. The samples were analyzed in atomic absorption spectrometer, and the results were evaluated through a statistical assesment, allowing the efficiency analysis of the effects and their interactions. The results showed percentages of extraction above 90% for the metals manganese, iron, chromium, calcium, barium and magnesium, and around 45% for metals lithium and potassium. The optimal point for the simultaneous removal of metals was calculated using statistical artifact multiple response function (MR). This calculation showed that the point of greatest extraction of metals occurs was the J point, with the composition [72% AP, 9% OP, 19% C/S], obtaining a global extraction percentage about 80%. Considering the aspects analyzed, the microemulsioned system has shown itself to be an effective alternative in the extraction of metals on synthetic produced water remediation

Purificação e caracterização parcial de uma B-D glicosidase de Artemia franciscana com ação sobre celobiose e lactose

-D-glucosidase (EC 3.2.1.21) is one of the most interesting glycosidases, especially for hydrolysis cellobiose releasing glucose, is last step degradation of cellulose. This function makes the -D-glucosidase is of great interest as a versatile industrial biocatalyst, being critical to various bio-treatment / biorefinery processes, such as bioethanol production. Hen in the report, a -D-glucosidase was extracts from protein extracted of the invertebrate marine Artemia franciscana was purified and characterized with a combination of precipitation with ammonium sulfate (0 - 30%, 30 to 50%, 50 to 80%), the fraction saturated in the range of 30 to 50% (called F-II) was applied in a molecular exclusion chromatography, in Sephacryl S-200, the fractions corresponding to the first peak of activity of -D-glucosidase were gathered and applied in a chromatography of ion exchange in Mono Q; the third peak this protein obtained chromatography, which coincides with the peak of activity of -D-glucosidase was held and applied in a gel filtration chromatography Superose 12 where the first peak protein, which has activity of -D-glucosidase was rechromatography on Superose 12. This enzyme is probably multimerica, consisting of three subunit molecular mass of 52.7 kDa (determined by SDS-PAGE) with native molecular mass of 157 kDa (determined by gel filtration chromatography on Superose 12 under the system FPLC). The enzyme was purified 44.09 times with a recovery of 1.01%. Using up p-nitrophenyl-β-D-glucopiranoside as substrate obtained a Km apparent of 0.229 mM and a Vmax of 1.109 mM.60min-1.mL-1mM. The optimum pH and optimum temperature of catalysis of the synthetic substrate were 5.0 and 45 °C, respectively. The activity of the -D-glucosidase was strongly, inhibited by silver nitrate and N- etylmaleimide, this inhibition indicates the involvement of radical sulfidrila the hydrolysis of synthetic substrate. The -D-glucosidase of Artemia franciscana presented degradativa action on celobiose, lactose and on the synthetic substrate -nitrophenyl-β-D-glucopiranoside indicating potential use of this enzyme in the industry mainly for the production of bioethanol (production of alcohol from the participating cellulose), and production hydrolysate milk (devoid of milk lactose)

Potencial terapêutico de um heparinóide isolado do invertebrado marinho Litopenaeus vannamei

The occurrence of bioactive compounds in marine organisms comes awaking the interest of the pharmaceutical industry. Heparin, a sulfated polysaccharide which presence was already identified in several marine invertebrates, is very attractive due its remarkable functional versatility. Besides to intervene in blood coagulation, this molecule has a great anti-inflammatory potential. However, its strong anticoagulant activity difficult the clinical exploitation of its anti-inflammatory properties. Thus, the aims of this work were to evaluate the effect of a heparin-like compound (heparinoid), isolated from the cephalotorax of the Litopenaeus vannamei shrimp, on the inflammatory response, hemostasia and synthesis of antithrombotic heparan sulfate by endothelial cells, besides studying some aspects concerning its structure. The purified heparinoid was structurally characterized following an analytical boarding, involving electrophoresis and chromatography. The structural analysis have shown that this compound possess a high content of glucuronic acid residues and disulfated disaccharide units. In contrast to mammalian heparin, the heparinoid was incapable to stimulate the synthesis of heparan sulfate by endothelial cells in the tested concentrations, beyond to show reduced anticoagulant activity and hemorrhagic effect. In a model of acute inflammation, the compound isolated from the shrimp reduced more than 50% of the cellular infiltration. Besides reduce the activity of MMP-9 and proMMP-2 of the peritoneal lavage of inflamed animals, the heparinoid also reduced the activity of MMP-9 secreted by activated human leukocytes. These results demonstrate the potential of heparinoid from L. vannamei to intervene in the inflammatory response. For possessing reduced anticoagulant activity and hemorrhagic effect, this compound can serve as a structural model to direct the development of more specific therapeutical agents to the treatment of inflammatory diseases