Use of real-time PCR to evaluate two DNA extraction methods from food

The DNA extraction is a critical step in Genetically Modified Organisms analysis based on real-time PCR. In this study, the CTAB and DNeasy methods provided good quality and quantity of DNA from the texturized soy protein, infant formula, and soy milk samples. Concerning the Certified Reference Material consisting of 5% Roundup Ready® soybean, neither method yielded DNA of good quality. However, the dilution test applied in the CTAB extracts showed no interference of inhibitory substances. The PCR efficiencies of lectin target amplification were not statistically different, and the coefficients of correlation (R²) demonstrated high degree of correlation between the copy numbers and the threshold cycle (Ct) values. ANOVA showed suitable adjustment of the regression and absence of significant linear deviations. The efficiencies of the p35S amplification were not statistically different, and all R² values using DNeasy extracts were above 0.98 with no significant linear deviations. Two out of three R² values using CTAB extracts were lower than 0.98, corresponding to lower degree of correlation, and the lack-of-fit test showed significant linear deviation in one run. The comparative analysis of the Ct values for the p35S and lectin targets demonstrated no statistical significant differences between the analytical curves of each target.

Sensory acceptability and physical stability evaluation of a prebiotic soy-based dessert developed with passion fruit juice

The aim of this study was to use a factorial design approach for developing a palatable and stable soy-based dessert with the addition of Soy Protein (SP), oligofructose, and Passion Fruit Juice (PFJ). Panelists (n = 50) used a seven-point hedonic scale to assess the overall liking, degree of liking of creaminess, taste, and color of the desserts. In addition, the samples were submitted to a preference ranking test in order to evaluate the products' preference. Water Holding Capacity (WHC) and backscattering (BS) measures were also determined to assess the physical stability of the trials. Sample F3 (35% PFJ and 2% SP) was the only one that presented a WHC index of 94.8%; moreover, none of the developed samples had synerisis after 72 hours of storage indicating adequate physical stability of the emulsion process. Samples F2 (25% PFJ, and 3.0% SP), F4 (35% PFJ, and 3.0% SP), and F5 (30% PFJ, and 2.5% SP) presented mean hedonic scores above 'slightly liked' for all sensory attributes. The acceptance index of samples varied from 62.50 to 88% showing the great sensory potential of such products.

Soursop juice stabilized with soy fractions: a rheologial approach

The potential use of soybean soluble polysaccharide (SSPS) as a stabilizer in acidic beverages was evaluated using rheological and stability studies. For this purpose, soy-based beverages were formulated with soy protein isolate (SPI) and soursop juice due to the low stability of this kind of dispersion. The influences of the concentrations of soybean soluble polysaccharide, calcium chloride, and soy protein isolate on the stability and rheology of soursop juice were evaluated using a factorial experimental design. Interactions between the concentrations of soybean soluble polysaccharide and soy protein isolate exerted a positive effect on the maximum Newtonian viscosity. The stability was positively influenced by the soybean soluble polysaccharide and soy protein isolate concentrations, but the interactions between soy protein isolate and CaCl2 also affected the sedimentation index. These results suggest that soybean soluble polysaccharide is effective in stabilizing fibers and proteins in acidic suspensions due to the increase in viscosity and steric effect caused by the formation of complexes between the soybean soluble polysaccharide and soy protein isolate.

Heat-treatment reduces anti-nutritional phytochemicals and maintains protein quality in genetically improved hulled soybean flour

The soybean is a protein source of high biological value. However, the presence of anti-nutritional factors affects its protein quality and limits the bioavailability of other nutrients. The effect of heat-treatment, 150 ºC for 30 minutes, on hulled and hull-less soybean flour from the cultivar UFVTN 105AP on urease, trypsin inhibitor activity, protein solubility, amino acid profile, and in vivo protein quality was investigated. The treatment reduced the trypsin inhibitor activity and urease, but it did not affect protein solubility. Protein Efficiency Coefficient (PER) values of the flours were similar, and the PER of the hull-less soybean flour did not differ from casein. The Net Protein Ratio (NPR) did not differ between the experimental groups. The True Digestibility (TD) of the flours did not differ, but both were lower in casein and the Protein Digestibility Corrected Amino Acid Score (PDCCAS) was lower than the TD, due to limited valine determined by the chemical score. Therefore, the flours showed reduced anti-nutritional phytochemicals and similar protein quality, and therefore the whole flours can be used as a source of high quality protein.

Detection of genetically modified maize events in Brazilian maize-derived food products

The Brazilian government has approved many transgenic maize lines for commercialization and has established a threshold of 1% for food labeling, which underscores need for monitoring programs. Thirty four samples including flours and different types of nacho chips were analyzed by conventional and real-time PCR in 2011 and 2012. The events MON810, Bt11, and TC1507 were detected in most of the samples, and NK603 was present only in the samples analyzed in 2012. The authorized lines GA21, T25, and the unauthorized Bt176 were not detected. All positive samples in the qualitative tests collected in 2011 showed a transgenic content higher than 1%, and none of them was correctly labeled. Regarding the samples collected in 2012, all positive samples were quantified higher than the threshold, and 47.0% were not correctly labeled. The overall results indicated that the major genetically modified organisms detected were MON810, TC1507, Bt11, and NK603 events. Some industries that had failed to label their products in 2011 started labeling them in 2012, demonstrating compliance with the current legislation observing the consumer rights. Although these results are encouraging, it has been clearly demonstrated the need for continuous monitoring programs to ensure consumers that food products are labeled properly.

Modeling and kinetic study of bio-ethanol production from soy protein concentrate by-product

Abstract The aim of this work was to evaluate a non-agitated process of bioethanol production from soybean molasses and the kinetic parameters of fermentation using a strain of Saccharomyces cerevisiae (ATCC® 2345). Kinetic experiment was conducted in medium with 30% (w v-1) of soluble solids without supplementation or pH adjustment. The maximum ethanol concentration was in 44 hours, the ethanol productivity was 0.946 g L-1 h-1, the yield over total initial sugars (Y1) was 47.87%, over consumed sugars (Y2) was 88.08% and specific cells production rate was 0.006 h-1. The mathematical polynomial was adjusted to the experimental data and provided very similar parameters of yield and productivity. Based in this study, for one ton of soybean molasses can be produced 103 kg of anhydrous bioethanol.

Detection of adventitious presence of genetically modified seeds in lots of non transgenic soybean

The difficulty on identifying, lack of segregation systems and absence of suitable standards for coexistence of non trangenic and transgenic soybean are contributing for contaminations that occur during productive system. The objective of this study was to evaluate the efficiency of two methods for detecting mixtures of seeds genetically modified (GM) into samples of non-GM soybean, in a way that seed lots can be assessed within the standards established by seed legislation. Two sizes of soybean samples (200 and 400 seeds), cv. BRSMG 810C (non-GM) and BRSMG 850GRR (GM), were assessed with four contamination levels (addition of GM seeds, for obtaining 0.0%, 0.5%, 1.0%, and 1.5% contamination), and two detection methods: immunoassay of lateral flux (ILF) and bioassay (pre-imbibition into 0.6% herbicide solution; 25 ºC; 16 h). The bioassay is efficient in detecting presence of GM seeds in seed samples of non-GM soybean, even for contamination lower than 1.0%, provided that seeds have high physiological quality. The ILF was positive, detecting the presence of target protein in contaminated samples, indicating test effectiveness. There was significant correlation between the two detection methods (r = 0.82; p < 0.0001). Sample size did not influence efficiency of the two methods in detecting presence of GM seeds.

LE || TIERS LIVRE || Des Faicts et Dicts || Heroïques du bon Pantagruel : || Composé par M. Fran. || Rabelais docteur || en Medi- || cine. || Reueu, & corrigé par l'Autheur, sus || la censure antique. || L'Autheur susdict || supplie les Lecteurs beneuoles, soy || reseruer a rire au soixante || & dixhuytiesme || Liure. || A Paris, || De l'imprimerie de Michel Fezandat, au mont || S. Hilaire, a l'hostel d'Albret. || 1552. Il Auec priuilege du Roy. In-8 de 160 f. inexactement chiffr. (le dernier coté 170), 3 f. non chiffr. et 1 f. blanc. — LE || QVART LIVRE || des faicts et || dicts Heroiques du bon || Pantagruel. || Composé par M. François Rabelais || docteur en Medicine. || A Paris, || De l'imprimerie de Michel Fezandat, au mont || S. Hilaire, a l'hostel d'Albret. || 1552. || Auec priuilege du Roy. — [A la fin de la Table :] Acheué d'imprimer le XXVIII. de || Ianuier M.D.LII [1552]. In-8 de 19 f. non chiffr., 144 f. chiffr. et 14 f. non chiffr. — Ensemble 2 vol. in-8, mar. citr., fil., dos ornés, tr. dor. (Anc. rel.). Rothschild 1514 (2)

Rabelais (François). Tiers Livre (1552)

Primary characterization of genes involved in the colony development of the insect pathogenic fungus Metarhizium anisopliae /

Strain improvement of the insect pathogenic fungus Metarhizium anisopUae is necessary to increase its virulence towards agricultural pests and thus improve its commercial efficacy. Nevertheless, the release of genetically modified conidia in crop fields may negatively affect the ecosystem. Controlling conidiation is a potential means of limiting the release of engineered strains since conidia are the infective propagules and the means of dispersal. The purpose of this study was to research the colony development of M. anisopUae to identify potential targets for genetic manipulation to control conidiation. Following Agrobacterium tumefaciem insertional mutagenesis, phenotypic mutants were characterized using Y-shaped adaptor dependent extension PCR. Four of 1 8 colony development recombinants had T-DNA flanking sequences with high homology to genes encoding known signaling pathway proteins that regulate pathogenesis and/or asexual development in filamentous fungi. Conidial density counts and insect bioassays suggested that a Serine/Threonine protein kinase COTl homolog is not essential for conidiation or virulence. Furthermore, a choline kinase homolog is important for conidiation, but not virulence. Finally, the regulator of G protein signaling CAG8 and a NADPH oxidase NoxA homolog are necessary for conidiation and virulence. These genes are candidates for further investigation into the regulatory pathways controlling conidiation to yield insight into promising gene targets for biocontrol strain improvement.

UNDERSTANDING THE RELATIONSHIP BETWEEN ONCOLYTIC AD5 DELETED E1b55KDA LYTIC INFECTION AND P53 IN MAMMALIAN CELLS

Adenoviruses are the most commonly used in the development of oncolytic therapy. Oncolytic adenoviruses are genetically modified to selectivity replicate in and kill tumor cells. The p53 molecule is a tumor suppressor protein that responds to viral infection through the activation of apoptosis, which is inhibited by adenovirus E1B55kDa protein leading to progressive viral lytic cycle. The non-specificity of replication has limited the use of wild type adenovirus in cancer therapy. This issue was resolved by using an E1b deleted Ad that can only replicate in cells with a deficiency in the p53 protein, a common feature of most cancer cells. Although demonstrating a moderate success rate, E1b55kDa deleted Ad has not been approved as a standard therapy for all cancer types. Several studies have revealed that E1b deleted Ad replication was independent of p53 status in the cell, as the virus replicated better in some p53 deficient cancers more than others. However, this mechanism has not been investigated deeply. Therefore, the objective of this study is to understand the relationship between p53 status, levels and functional activity, and oncolytic Ad5dlE1b55kDa replication efficiency. Firstly, five transient p53 expression vectors that contain different regulatory elements were engineered and then evaluated in H1299, HEK293 and HeLa cell lines. Data indicated that vector that contains the MARs and HPRE regulatory elements achieved the highest stability of p53 expression. Secondly, we used these vectors to examine the effect of various p53 expression levels on the replication efficiency of oncolytic Ad5dlE1b55kDa. We found that the level of p53 in the cell had an insignificant effect on the oncolytic viruses’ replication. However, the functional activity of p53 had a significant effect on its replication, as Ad5dlE1b55kDa was shown to have selective activity in H1299 cells (p53-null). In contrast, a decrease in viral replication was found in HeLa cells (p53-positive). Finally, the effect of p53’s functional activity on the replication efficiency of oncolytic Ad5dlE1b55kDa was examined. Viral growth was evaluated in H1299 cells expressing number of p53 mutants. P53-R175H mutant successfully rescued viral growth by allowing the virus to exert its mechanism of selectivity. The mechanism entailed deregulating the expression of specific genes, cell cycle and apoptosis, in the p53 pathway to promote its production leading to efficient oncolytic effect. These results confirmed that oncolytic Ad5dlE1b55kDa sensitivity is mutation-type specific. Therefore, before it is applied clinically as cancer therapy for p53 deficient tumors, the type of p53 mutation must be determined for efficient antitumor effect.

Esta mujer que está aquí no soy yo: el mundo de una neurosis de histeria en el retorno de lo reprimido manifestado en una conversión.

Tesis (Maestría en Psicología Clínica con orientación Psicoanalítica) UANL, 2014.

Principles for Incorporating Farmers in the Ethical Assessment of Genetically Modified Crops

A current advance within the agricultural industry is the use of genetic engineering to produce novel crops for food production. This technology raises questions about how societies should position themselves with respect to genetically modified (GM) crop development and implementation; namely, how should the potentials and risks of this technology be evaluated? We argue that current methods to evaluate the risks and benefits of GM crops are inadequate and not conducive to the strategic development of this technology, where a way to ameliorate technology assessments for GM crops is to include farmers in the research process of evaluating these crops prior to their commercialization. However, particularities concerning the ethical status of such research require special consideration and vigilance. For example, in such technology assessment initiatives, farmers would occupy both the roles of research participant and research investigator. Other particularities surface due to factors related to the nature of GM crops. These particularities are examined with reference to concepts drawn from the field of research ethics, namely informed consent, compensatory decisions, and issues of participant inclusion/exclusion.

Characterization of a Canadian mink H3N2 influenza A virus isolate genetically related to triple reassortant swine influenza virus.

In 2007, an H3N2 influenza A virus was isolated from Canadian mink. This virus was found to be phylogenetically related to a triple reassortant influenza virus which emerged in Canadian swine in 2005, but it is antigenically distinct. The transmission of the virus from swine to mink seems to have occurred following the feeding of animals with a ration composed of uncooked meat by-products of swine obtained from slaughterhouse facilities. Serological analyses suggest that the mink influenza virus does not circulate in the swine population. Presently, the prevalence of influenza virus in Canadian farmed and wild mink populations is unknown. The natural occurrence of influenza virus infection in mink with the presence of clinical signs is a rare event that deserves to be reported.

Sí, señor, soy colombiana de pura cepa, et toi? : las formas de tratamiento del español colombiano en Montreal

Le présent travail décrit –pour la première fois– l’état actuel de la langue espagnole parlée par la communauté colombienne à Montréal sur les formes de s’adresser aux autres en langage pronominal ou nominal et la courtoisie verbale. Dans le but de réaliser cette étude, un travail de terrain a été effectué à l’aide d’un questionnaire et des entrevues orales semi-dirigées adressés à 30 informateurs. L’analyse des données, nous a permis d’établir quelques premières comparaisons entre la façon de parler des habitants de la Colombie et des Colombiens résidant à Montréal et d’identifier quelques-uns des changements linguistiques principaux dans cette communauté parlante, notamment, les variations reliées aux formes de s’adresser aux autres et aux actes de courtoisie affectés par l’influence du français et de l’anglais. L’analyse effectuée tient compte autant les aspects linguistiques, pragmatiques et sociaux que les attitudes linguistiques des interviewés. De cette façon, les résultats mettent en lumière une nouvelle description sur la dynamique de l’usage des formes de s’adresser aux autres de locuteurs originaires de trois zones dialectales représentatives de la Colombie : la région andine orientale, la région andine occidentale et la zone côtière du Caraïbe. Ensemble avec d’autres études précédentes sur la formes de s’adresser aux autres, ce travail constitue une meilleure compréhension de la réalité linguistique de l’espagnol des Colombiens unilingues, bilingues et trilingues.