946 resultados para solution-based DNA extraction
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Sexually transmitted diseases (STDs) are among the largest public health problems, especially in developing countries. The acquisition of these infections during early sexual activity is common and many infections have a benign course. However, in some pathogens remain in the state of latency can be reactivated and cause productive infection that may progress to severe forms. In addition, some of them are transmitted vertically resulting in congenital infection, causing immediate damage or long-term child. The classic risk factors for sexually transmitted agents are: early onset of sexual and reproductive health, multiple sexual partners throughout life, use of oral contraceptives and co-infections with different pathogens. We present the results of a cross-sectional study aimed to estimate the prevalence of genital infection by human papillomavirus (HPV), Herpes simplex virus (HSV) and Chlamydia trachomatis (CT) in a segment of the female population of the metropolitan area Christmas, among those who enrolled voluntarily sought, Basic Health Units for the examination of cancer screening cervix in the period 2008 to 2010. All participants, a total of 261 women answered a standard questionnaire by which identified the socio-demographic characteristics, classical risk factors for STDs, reproductive and sexual activity and smoking. Of each patient were obtained two samples, one for the completion of the Pap test for detection of cellular changes and the other processed for DNA extraction and analyzed by PCR (polymerase chain reaction) to detect the three pathogens studied. The population of the study was composed of sexually active women aged between 13 and 79 years, mean 38.7 years, most of them being married, low education levels and low incomes. The majority (87%) had normal results on cytology and only 2.7% had low-grade cytological abnormalities. Prevalence rates were 37.9% for HPV, 4.6% for CT and 26% for HSV. HPV prevalence was higher in women under 25, unmarried and in those who had multiple sexual partners. Women with simultaneous infection by HSV-1 and 2 had higher prevalence of HPV infection. The prevalence of HSV infection showed no association whatsoever with the risk factors analyzed and HSV-1 was the predominant type among the cases of genital HSV infection. The overall prevalence of C. Trachomatis was relatively low, thus providing greater value in younger women aged less than or equal to 20 years
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Herpes simplex is a virus that can be transmitted sexually and is potentially associated with vertical transmission. This study evaluated the prevalence of genital infection by herpes simplex virus (HSV) types 1 and 2 in pregnant and nonpregnant care in the city of Natal / RN, including a total of 222 women, 92 pregnant and 130 nonpregnant. The participants answered a questionnaire to obtain data and socio-demographic characteristics, as well as potential risk factors for sexually transmitted diseases. After the interview, we collected two cervical specimens, one for the Pap test and the other for DNA extraction and analyzed by polymerase chain reaction (PCR) to detect both virus serotypes. Then the women underwent a clinical examination by colposcopy. For statistical analysis, we used the chi-square and logistic regression by SSPS 17.0 Statistic. Most women were up to 30 years of age, nonwhite ethnicity, married, elementary education, family income below the poverty level; initiated sexual activity with age up to 18 years; had more than one sexual partner lifelong and was not pregnant, but has had at least one child. The HSV-1 showed a prevalence of 26.1% among pregnant women and 30.0% in non-pregnant women. While HSV-2 prevalence was found with 10.9% and 19.2% in pregnant and nonpregnant women, respectively. The largest proportion of morphological changes of the uterine cervix was detected among nonpregnant women, both in cytology and in colposcopy. The women were nonwhite ethnicity, married, became pregnant aged less than or equal to 18 years and who had one to two pregnancies had a lower risk of acquiring genital HSV infection. There was a high prevalence of genital HSV infection, HSV-1 is more prevalent than HSV-2. No association was found between morphological changes of the uterine cervix and the presence of the virus in pregnant and nonpregnant women, nor between genital HSV infection and the classic risk factors for sexually transmitted diseases
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The congenital facial clefts are characterized by incomplete formation of the structures that separate the oral and nasal cavity. It is known that several environmental and genetic factors are involved in its development, among these, polymorphisms associated with folic acid metabolism have been investigated. In this sense, the objective was to observe the frequency of polymorphisms C677T and A1298C methylenetetrahydrofolate reductase gene (MTHFR), methionine synthase A2756G of (MTR), A66G of methionine synthase reductase (MTRR) A80G and the reduced folate carrier (RFC1) in patients with non-syndromic oral clefts, trying to match them with their development. Methods: We studied 140 patients with non-syndromic oral clefts and their mothers and 175 control subjects with their mothers, who underwent a questionnaire to obtain family information. Were collecting blood for DNA extraction from patients and their mothers to identify the genotypes of both by PCRRFLP, in addition to carrying out the determination of glucose, AST, ALT and serum creatinine, folic acid and vitamin B12 Serum and plasma homocysteine, and the hemogram. Results: Most patients have cleft lip and palate (55.8%), followed by isolated cleft palate (24.2%) and cleft lip (20%). Regarding gender, 62% of patients were male and 48% female and, after subdivision of the type of screwdriver according to sex was found a prevalence of males in the cracks of the type lip and palate (69 %) and lip (69.2%) and in the case of cleft palate was a female predominance (59%). The average concentration of serum folate in the group of mothers of cleft patients was significantly lower (13.8 ± 2.4 ng / mL) compared with the group of mothers of control subjects (18.8 ± 3.4 ng / mL) This was also observed for the group of cleft children as compared to controls, the dosage of folic acid had a significant difference with values of 15.6 ± 0.6 (ng / mL) and 17.9 ± 0.6 (ng / mL), respectively. For the biochemical measurements of glucose, AST, ALT and creatinine were not statistically different, nor was observed for haematological parameters performed. In assessing the frequency of polymorphisms C677T and A1298C MTHFR, A2756G MTR, MTRR A66G and A80G of the RFC1 there was no statistically significant difference in genotype distribution between cases and controls both for mothers and in the cleft. Conclusion: Although not observed association of polymorphisms with the development of cracks, the decrease in serum folate in the group of cleft patients and their mothers may reflect a disturbance in the metabolism of this metabolite, necessitating further studies such as studies methylation and expression to further elucidate the involvement of folate in the development of oral clefts
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O flamboyanzinho (Caesalpinia pulcherrima, Fabaceae) é muito utilizado como cerca-viva e na arborização de ruas, parques e jardins. de florescimento exuberante, suas flores podem apresentar coloração rosa, laranja ou amarela, conforme a variedade. Como características florais são essenciais para a definição do valor comercial de espécies ornamentais, o objetivo principal do trabalho foi verificar a existência de polimorfismo entre plantas de C. pulcherrima, que produzem flores de diferentes colorações, por marcadores moleculares RAPD. Foram escolhidas aleatoriamente 30 plantas adultas, cultivadas no município de Jaboticabal, Estado de São Paulo, para a retirada de amostras foliares para a extração de DNA. Dentre essas matrizes, 20 possuem flores alaranjadas, oito, flores amarelas, e apenas duas produzem flores de coloração rosa. Dos 140 primers de RAPD avaliados, 94 foram capazes de ampliar fragmentos definidos, gerando 246 bandas, das quais se observou 100% de bandas monomórficas, indicando que nenhum dos primers utilizados detectou polimorfismo entre os tratamentos. Concluiu-se que a técnica para detecção de polimorfismo por marcadores RAPD não foi eficiente, e que existe a necessidade de se testarem marcadores moleculares mais específicos. Além disso, a característica morfológica cor de flor é, possivelmente, controlada por vários genes ou pela combinação deles.
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This study aimed to characterize molecular of 13 accessions of Psidium spp. (Myrtaceae) that was been identified for the reaction to rootknot guava nematode. The DNA extraction of the samples was carried according to the protocol of Shillito & Saul (1988). The molecular markers type fAFLP, were obtained from fAFLP Regular Plant Genomes Fingerprinting Kit' (Applied Biosystems from Brasil Ltda.) and were tested 24 selectives combinations of primers, of which 18 showed amplification that produced 272 polymorphic markers. To the analysis of the markers were employed the softwares GeneScan (ABI Prism versao 1.0) and Genotyper (ABI Prism version 1.03), and the data collected were transformed into a binary matrix that was analyzed in the software PAUP (Phylogenetic Analysis Using Parcimony - version 3.01). Were calculated genetic distance index intra and interespecific between the genotipes. It was found that the AFLP markers were efficient in the discrimination between accessions, as well as in showing genetic similarity among accessions identified as resistant to the nematode Meloidogyne enterolobii, which could be discussed in the future.
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
A nested-PCR assay for detection of Xylella fastidiosa in citrus plants and sharpshooter leafhoppers
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Aims: Detection of Xylella fastidiosa in citrus plants and insect vectors.Methods and Results: Chelex 100 resin matrix was successfully standardized allowing a fast DNA extraction of X. fastidiosa. An amplicon of 500 bp was observed in samples of citrus leaf and citrus xylem extract, with and without symptoms of citrus variegated chlorosis, using PCR with a specific primer set indicating the presence of X. fastidiosa. The addition of insoluble acid-washed polyvinylpyrrolidone (PVPP) prior to DNA extraction of insect samples using Chelex 100 resin together with nested-PCR permitted the detection of X. fastidiosa within sharpshooter heads with great sensitivity. It was possible to detect up to two bacteria per reaction. From 250 sharpshooter samples comprising four species (Dilobopterus costalimai, Oncometopia facialis, Bucephalogonia xanthopis and Acrogonia sp.), 87 individuals showed positive results for X. fastidiosa in a nested-PCR assay.Conclusions: the use of Chelex 100 resin allowed a fast and efficient DNA extraction to be used in the detection of X. fastidiosa in citrus plants and insect vectors by PCR and nested-PCR assays, respectively.Significance and Impact of the study: the employment of efficient and sensitive methods to detect X. fastidiosa in citrus plants and insect vectors will greatly assist epidemiological studies.
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The goal of this work is to propose a SLAM (Simultaneous Localization and Mapping) solution based on Extended Kalman Filter (EKF) in order to make possible a robot navigates along the environment using information from odometry and pre-existing lines on the floor. Initially, a segmentation step is necessary to classify parts of the image in floor or non floor . Then the image processing identifies floor lines and the parameters of these lines are mapped to world using a homography matrix. Finally, the identified lines are used in SLAM as landmarks in order to build a feature map. In parallel, using the corrected robot pose, the uncertainty about the pose and also the part non floor of the image, it is possible to build an occupancy grid map and generate a metric map with the obstacle s description. A greater autonomy for the robot is attained by using the two types of obtained map (the metric map and the features map). Thus, it is possible to run path planning tasks in parallel with localization and mapping. Practical results are presented to validate the proposal
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This work intends to show a new and few explored SLAM approach inside the simultaneous localization and mapping problem (SLAM). The purpose is to put a mobile robot to work in an indoor environment. The robot should map the environment and localize itself in the map. The robot used in the tests has an upward camera and encoders on the wheels. The landmarks in this built map are light splotches on the images of the camera caused by luminaries on the ceil. This work develops a solution based on Extended Kalman Filter to the SLAM problem using a developed observation model. Several developed tests and softwares to accomplish the SLAM experiments are shown in details
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Vegetables drying plays an important role in the field of food dehydration, being a very old practice that was originated from sun drying items of food in order to preserve them to be consumed during the periods of scarcity. One of these vegetables is the tomato, that was originally grown in South America. Tomatoes are easily perishable after being picked up from the tree and this makes the process of tomato dehydration a challenge due to the high amount of water (95%) contained in them. The present research work was mainly intended to develop alternative processes for tomato conservation, by drying slices of skinned and unskinned tomatoes in the in natura form or in the osmotically pre-dehydrated form. Firstly, the best conditions of the osmotic pre-dehydration process were defined including temperature, immersion time and concentration of the osmotic solution, based on the results of water loss, solids gain and weigh reduction of the pre-dehydration tomatoes at different processing conditions. The osmotic solution used was made up of NaCl (5 and 10%) and sucrose (25 and 35%) at different combinations. For a fixed conditions of osmotic pre-dehydration, the drying tests of the pre-processed and in natura tomatoes were carried out in a stove with air circulation and a convective dryer with trays, at two levels of temperature. The sensorial analysis of the osmotically pre-treated and unskinned dehydrated tomatoes was carried out as well as a study on the their shelf-live. The results obtained showed that the drying of the tomatoes took place as a result of the internal control of the water transport, and did not show a constant rate, while two distinct periods of the decreasing phase were observed. The osmotic pre treatment substancially reduced the initial amount of humidity in the tomatoes, thus reducing the necessary time for the product to attain levels of intermediate humidity. The impermeability of the tomato skin was identified as well as the unfavorable influence of the pre-treatment on the unskinned tomatoes, whose solid gain brought about a decrease in the water activity with subsequent reduction of the drying rate. Despite the various simplifications carried out during the development of this study, the proposed diffusive model adjusted to the experimental data satisfactorily, thus making it possible to determine the effective coefficients of diffusion, whose results were consistent and compatible with those found in the current literature. Concerning the higher rates of evaporation and the lowest processing time, the best results were obtained in the drying of the unskinned, in natura tomatoes and of the skinned, pre-dehydrated tomatoes, at 60ºC, both processed in the convective drier. The results of the sensorial analysis of the unskinned and pre-treated product did not prove to be satisfactory. Regarding the shelf-live of the tomatoes, for a period of 45 days, no physicochemical or microbiological alteration of the product was noted
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In the area of food dehydration, drying of vegetables has a very representative position, it has the objective to preserve the surplus of crops and began with sun drying. Among the vegetable is the carrot, which had its origin in Southeast Asia and in Brazil is a vegetable cultivated enough. The principal objective of this works is to find alternative ways for the conservation of carrot slices by osmotic dehydration with additional drying in heart. Were initially defined the best conditions of pre-osmotic dehydration (temperature, immersion time, type of osmotic solution) based on the results of humidity loss, solid gain, weight reduction and efficiency ratio of predehydrated carrots slices. The osmotic solutions used were composed by NaCl (10%) and sucrose (50 ° Brix) named DO1 and sucrose (50 ° Brix) called DO2. Was made experiment of pre-osmotic dehydration of carrot slices in two temperature levels, with complementary drying in heart with air circulation at 70 º C. Sensory analysis was performed and the study of slices dehydration osmotically and the slices without osmotic treatment. The best results were obtained with the solution DO1 60°C with immersion time of 60 min. The drying of carrot slices presented period of constant rate and decreasing rate. The osmotic pre-treatment reduced the initial humidity of carrot slices, reducing the time to the product to reach the same humidity content. Fick's model, considering the shrinkage, and the Page s model, adapt satisfactorily to experimental datas, allowing the determination of effective diffusion coefficients, consistent with the references. The results of sensory analysis of dry product, showed greater acceptance of sliced carrots with osmotic treatment
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The diesel combustion form sulfur oxides that can be discharged into the atmosphere as particulates and primary pollutants, SO2and SO3, causing great damage to the environment and to human health. These products can be transformed into acids in the combustion chamber, causing damage to the engines. The worldwide concern with a clean and healthy environment has led to more restrictive laws and regulations regulating the emission levels of pollutants in the air, establishing sulfur levels increasingly low on fuels. The conventional methods for sulfur removal from diesel are expensive and do not produce a zero-level sulfur fuel. This work aims to develop new methods of removing sulfur from commercial diesel using surfactants and microemulsion systems. Its main purpose is to create new technologies and add economic viability to the process. First, a preliminary study using as extracting agent a Winsor I microemulsion system with dodecyl ammonium chloride (DDACl) and nonyl phenol ethoxylated (RNX95) as surfactant was performed to choose the surfactant. The RNX95 was chosen to be used as surfactant in microemulsioned systems for adsorbent surface modification and as an extracting agent in liquid-liquid extraction. Vermiculite was evaluated as adsorbent. The microemulsion systems applied for vermiculite surface modification were composed by RNX95 (surfactant), n-butanol (cosurfactant), n-hexane (oil phase), and different aqueous phases, including: distilled water (aqueous phase),20ppm CaCl2solution, and 1500ppm CaCl2solution. Batch and column adsorption tests were carried out to estimate the ability of vermiculite to adsorb sulfur from diesel. It was used in the experiments a commercial diesel fuel with 1,233ppm initial sulfur concentration. The batch experiments were performed according to a factorial design (23). Two experimental sets were accomplished: the first one applying 1:2 vermiculite to diesel ratio and the second one using 1:5 vermiculite to diesel ratio. It was evaluated the effects of temperature (25°C and 60°C), concentration of CaCl2in the aqueous phase (20ppm and 1500ppm), and vermiculite granule size (65 and 100 mesh). The experimental response was the ability of vermiculite to adsorb sulfur. The best results for both 1:5 and 1:2 ratios were obtained using 60°C, 1500ppm CaCl2solution, and 65 mesh. The best adsorption capacities for 1:5 ratio and for 1:2 ratio were 4.24 mg sulfur/g adsorbent and 2.87 mg sulfur/g adsorbent, respectively. It was verified that the most significant factor was the concentration of the CaCl2 solution. Liquid-liquid extraction experiments were performed in two and six steps using the same surfactant to diesel ratio. It was obtained 46.8% sulfur removal in two-step experiment and 73.15% in six-step one. An alternative study, for comparison purposes, was made using bentonite and diatomite asadsorbents. The batch experiments were done using microemulsion systems with the same aqueous phases evaluated in vermiculite study and also 20ppm and 1500 ppm BaCl2 solutions. For bentonite, the best adsorption capacity was 7.53mg sulfur/g adsorbent with distilled water as aqueous phase of the microemulsion system and for diatomite the best result was 17.04 mg sulfur/g adsorbent using a 20ppm CaCl2solution. The accomplishment of this study allowed us to conclude that, among the alternatives tested, the adsorption process using adsorbents modified by microemulsion systems was considered the best process for sulfur removal from diesel fuel. The optimization and scale upof the process constitutes a viable alternative to achieve the needs of the market
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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
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Most methods employed to estimate the height of waves generated by the wind require the surface length over which the wind blows. The choice of method depends on the nature of the water body, being applicable to ocean areas or interior water bodies, such as bays, lakes and reservoirs. One of the usual solutions has been the direct usage of the method validated in ocean waters for interior waters, therefore, not taking into consideration the effect of the restriction imposed by the shores. Nevertheless, the excessive quantity of operations of the method applied to interior water bodies (where the shore is a restraint condition) may not assure a satisfactory precision degree, unless an accurate enough graphic base of the shore with the addition of the operator's subjectivity is used. Thus, this scientific community brings this discussion to light, proposing a classic solution based on the application of the adequate method to interior waters (Saville et al., 1954) via automatized processing. Therefore, a program in AutoLISP, a computational language, has been developed. The application of the program has determined the maximum wind fetches in Ilha Solteira, state of São Paulo, reservoir as being between 9.5 and 12.5 km, in contrast with a previous study which has predicted far longer fetches (factor of three).
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This study aimed to analyze the effect of a saline solution on growth and chemical composition of Atriplex nummularia, shrubby plant, absorbing salts used in the diet of animals and the management of water and saline soils. These plant seedlings were planted and grown in a reserved area at the Federal University of Rio Grande do Norte. The plantation was divided into two blocks, in which one of them was irrigated with saline solution with a concentration of 2840 mgL-1 of NaCl and the second group was irrigated with drinking water. After six months, the plants were collected, harvested and divided into three parts: leaf, thin and thick stem. Monthly, dimension measurements were carried out for cataloging the growth of Atriplex. Ion Chromatography (IC) and Optical Emission Spectroscopy Inductively Coupled Plasma (ICP-OES) were used to analyze the chemical composition of the partition plant parts. The results of these analyses revealed that an absorption process of anions and cations by Atriplex nummularia plant during its growth was achieved, in particular by a higher concentration of sodium and chloride ions. Scanning electron microscopy images showed and confirmed the presence of small crystals on the leaf surface. Electrical conductivity and pH measurements of the aerial parts of the plant were carried out and these results showed that the leaf is the plant part where there is a largest concentration of ions. In addition, measurements of specific surface were obtained from irrigated plants with saline solution, achieving higher surface area, in all cases. Plant dimensions obtained monthly showed that the plants irrigated with water grew 5% more than those plants irrigated with saline solution. Based on results obtained, Atriplex plant showed a higher potential to survive and adapt to environments (aquatic or geological) with high levels of salinity and this property can be used as a tool for removing salts/metals from industrial contaminated soils and effluents.