EGCG Treatment on Ts65Dn Mice Suggests a Possible Correlation in Cognitive Development Deficit Reduction

poster abstractDown syndrome (DS) is caused by trisomy of human chromosome 21 (Ts21), affecting 1 in 700 live births. Ts21 results in about 80 phenotypes of which intellectual disability (ID) is one of the most debilitating. DYRK1A, found in 3 copies in individuals with Ts21 has been linked to alterations in morphology and function of the brain resulting in ID. Epigallocatechin-3-gallate (EGCG), a specific inhibitor of Dyrk1a activity has been hypothesized as a possible treatment for the overexpression of this gene, reducing the deficits caused by Dryk1a. Using the Ts65Dn mouse model, we examined the effects on hippocampal dependent learning and memory in the novel object recognition task (NOR) using mice of 3-6 weeks of age (adolescent mice). They were given free access to EGCG (0.124 mg/mL) in their drinking water for 21 days. They were then tested for cognitive improvement through NOR. Ts65Dn and control mice (treated and untreated) were subjected to 3 days of testing with 15 minute sessions per day consisting of habituation, exposure, and test day. All procedures were recorded and analyzed to determine time spent exploring novel object in relation to familiar. Our current results suggest that s65Dn mice do not spend as much time exploring the novel object as euploid mice and there exists a genotype effect, but treatment is not correcting the learning and memory deficit. We hypothesize that continuous EGCG treatment may be needed in order to see cognitive deficit reduction in adolescent mice

Effects of 50 mg/kg EGCG Treatment of Ts65Dn Down Syndrome Mice on Novel Object Recognition

poster abstractDown syndrome (DS) is caused by trisomy of chromosome 21, and affects 1/700 live births. DYRK1A, a gene found in three copies in humans with DS and Ts65Dn DS mice, has been linked to alterations in morphology and function of the brain resulting in cognitive impairment. Epigallocatechin-3-gallate (EGCG), an inhibitor of DYRK1A activity, has been proposed as a possible treatment. Using the Ts65Dn DS mouse model, we examined the effects of EGCG treatment on on hippocampal dependent learning and memory using a novel object recognition task (NOR). A previous study analyzing the effects of EGCG at a concentration 30mg/kg/day showed that there was no genotype or treatment effect in the NOR task when treatment is continuous through testing. In this study, the mice were given 50 mg/kg/day EGCG or water via their drinking water starting at 3 weeks of age. The mice were handled two days before testing and then underwent a series of behavioral tests including NOR. They underwent testing at 3 weeks and 7 weeks of treatment. Treatment was continuous throughout behavioral testing. NOR consists of a box with the objects placed diagonally from each other. The mice underwent 3 days of testing with 15 minute sessions per day consisting of habituation, exposure, and test day, all of which were recorded and analyzed to determine time spent exploring novel object in relation to familiar. The amount of time spent at each object was scored by three independent scorers, blind to genotype and treatment. We observed no genotype or treatment effect at either the 3 or 7 week test results, which is consistent with our past results. A higher dose, along with a more sensitive test of recognition memory, may be needed in order to show a treatment effect on the Ts65Dn mice

Effects of EGCG Treatment of Ts65Dn Down Syndrome Mice on a Balance Beam Task

poster abstractDown syndrome (DS) is caused by trisomy of chromosome 21, and affects 1/700 live births. DS results in about 80 clinical phenotypes, including cognitive impairment. DYRK1A, a chromosome 21 gene, has been linked to alterations in morphology and function of the brain resulting in cognitive impairment. Epigallocatechin-3-gallate (EGCG), an inhibitor of DYRK1A activity, has been proposed as a possible treatment for cognitive deficits seen in individuals with DS. Using the Ts65Dn DS mouse model, we examined the effects of EGCG treatment on cerebellum dependent tasks using a balance beam test. We hypothesized that treatment with EGCG would improve Ts65Dn performance on the balance beam. In a first experiment, mice were given a dose of ~30 mg/kg/day EGCG, which showed no significant improvement in the balance beam task. In a second experiment, mice were given a dose of 100 mg/kg/day EGCG or water (control) starting at 3 weeks of age. The mice were handled two days before testing and then underwent a series of behavioral tasks including the balance beam test. The mice traversed three beams of differing widths (12, 9 and 6 mm), and three consecutive trials for each were recorded for further analysis. The balance beam recordings were scored by three independent scorers, blind to genotype and treatment, and the number of hind paw slips for each trial were scored. Our preliminary results indicate that the Ts65Dn mice are impaired at this task and have more hind paw slips compared to euploid controls. A larger number of animals should help to distinguish any differences in Ts65Dn mice due to EGCG treatment

Esterase activities and biochemical properties of lactic acid bacteria isolated from goat´s milk cheese in Argentina

Twenty-two lactic acid bacteria (LAB) strains isolated from Argentinean goat dairy products were evaluated for its biochemical properties and esterase activities relevant to flavor development. Streptococcus thermophiles (UNSE314), Lactobacillus (L.) delbrueckii subsp. bulgaricus (UNSE309), L. rhamnosus (UNSE308), L. plantarum (UNSE287, UNSE316, UNSE317) and Pediococcus pentosaceus (UNSE315) strains presented high acidifying activity. All strains tested metabolized citrate and produced diacetyl-acetoin in goat milk. Based on these results, ten strains with the best performance in diverse technological properties were selected to determine esterolytic activity. In all evaluated strains, esterase specific activity (ESA) was detected on a-naphthyl (a-NA) acetate and B-naphthyl (B-NA) acetate, propionate, caprylate and a-NA butyrate. No activity was detected on B-NA laurate. The highest values were detected when using a-NA instead of B-NA derivatives as substrate. In Pediococcus strains, wide variability in ESA were observed, which were species- and strain-specific. These results allow us to select strains with biochemical properties and esterase activities to design starter and adjunct cultures that contribute to flavor development during cheese ripening, thus preserving the typical organoleptic characteristics of Argentinean goat cheeses. Fil: Taboada, Natalia Verónica. Universidad Nacional de Santiago del Estero. Facultad de Agronomia y Agroindustrias. Instituto de Cs.y Tecnologías Alimentarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Lopez Alzogaray, Maria Soledad. Universidad Nacional de Santiago del Estero. Facultad de Agronomia y Agroindustrias. Instituto de Cs.y Tecnologías Alimentarias; ArgentinaFil: Abeijon Mukdsi, Maria Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentina. Universidad del Norte Santo Tomás de Aquino; ArgentinaFil: Medina, Roxana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentina. Universidad Nacional de Tucumán. Facultad de Agronomía y Zootecnia; Argentin

Fast assembly of non-thiolated DNA on gold surface at lower pH

The final publication is available at Elsevier via http://dx.doi.org/10.1016/j.jcis.2013.08.043 © 2013. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/In a typical protocol for attaching DNA to a gold electrode, thiolated DNA is incubated with the electrode at neutral pH overnight. Here we report fast adsorption of non-thiolated DNA oligomers on gold electrodes at acidic pH (i.e., pH ∼3.0). The peak-to-peak potential difference and the redox peak currents in typical cyclic voltammetry of [Fe(CN)6]3− are investigated to monitor the attachment. Compared with incubation at neutral pH, the lower pH can significantly promote the adsorption processes, enabling efficient adsorption even in 30 min. The adsorption rate is DNA concentration-dependent, while the ionic strength shows no influence. Moreover, the adsorption is base-discriminative, with a preferred order of A > C ≫ G, T, which is attributed to the protonation of A and C at low pH and their higher binding affinity to gold surface. The immobilized DNA is functional and can hybridize with its complementary DNA but not a random DNA. This work is promising to provide a useful time-saving strategy for DNA assembly on gold electrodes, allowing fast fabrication of DNA-based biosensors and devices.National Natural Science Foundation of China || 20905012 University of Waterloo || Canadian Foundation for Innovation || Ontario Ministry of Research & Innovation || Natural Sciences and Engineering Research Council Americas Program |

Glutathione-s-transferase modified electrodes for detecting anticancer drugs

The final publication is available at Elsevier via http://dx.doi.org/10.1016/j.bios.2014.02.070. © 2014. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/With the fast growth of cancer research, new analytical methods are needed to measure anticancer drugs. This is usually accomplished by using sophisticated analytical instruments. Biosensors are attractive candidates for measuring anticancer drugs, but currently few biosensors can achieve this goal. In particular, it is challenging to have a general method to monitor various types of anticancer drugs with different structures. In this work, a biosensor was developed to detect anticancer drugs by modifying carbon paste electrodes with glutathione-s-transferase (GST) enzymes. GST is widely studied in the metabolism of xenobiotics and is a major contributing factor in resistance to anticancer drugs. The measurement of anticancer drugs is based on competition between 1-chloro-2,4-dinitrobenzene (CDNB) and the drugs for the GST enzyme in the electrochemical potential at 0.1 V vs. Ag/AgCl by square wave voltammetry (SWV) or using a colorimetric method. The sensor shows a detection limit of 8.8 μM cisplatin and exhibits relatively long life time in daily measurements.University of Waterloo || Natural Sciences and Engineering Research Council || Emerging Leaders in the Americas Program |

Avaliação das variáveis físico-químicas de um reservatório subtropical no noroeste da Argentina

AIM: The Escaba dam is located in the south of the Tucumán province, Argentina, at 650 m above sea level. It has an extension of 541 ha. and a depth of 65 m and its tributaries are the Chavarría, Las Moras, El Chorro and Singuil rivers. The climate is mild with dry winters and rainy summers. The objective of this study was to characterize physicochemical parameters in the limnetic zone of the dam and the mouths of the tributaries to determine the water quality. METHODS: Seasonal sampling was carried out between August 2010 and May 2012. Temperature, transparency, pH and electrical conductivity were field measured, whereas dissolved oxygen, biochemical oxygen demand (BOD5), major ion constituents and nitrogen and phosphate compounds were analyzed at the laboratory. RESULTS: The water was classified as sodium-calcium-bicarbonate with neutral to alkaline pH, and thermal stratification during spring and summer. The water assayed was well oxygenated except for the bottom of the limnetic zone during the summer months. Lowest transparency was measured in the El Chorro River in November 2011 (0.12 m) and highest degree of transparency in the Singuil River during the winter of 2010 (4.1 m). The waters assayed showed weak mineralization with conductivities between 83 and 218 µS.cm-1. Maximum BOD5 value (183 mg.L-1) was measured in the Singuil River in spring 2010. Highest values for the different nitrogen compounds were as follows: 7 mg NO3-.L-1 at the bottom of the limnetic zone in August 2010, 0.07 mg NO2-.L-1 in the Las Moras River in May 2011 and 1.8 mg NH4+.L-1 in the Chavarría River in March 2011. During the summer of 2012 orthophosphate reached a value of 0.22 mg.L-1 at the bottom of the limnetic zone. The TN/TP ratio revealed that phosphate was generally the limiting factor and rarely nitrogen. CONCLUSIONS: Considering the TN, TP and transparency parameters the ecosystem was classified as hypertrophic. PCA allowed a seasonal differentiation of the sites, and components 1 and 2 classified the samples according to nutrient gradient, dissolved oxygen, BOD5 and temperature.OBJETIVO: A represa Escaba está localizada ao sul da província de Tucumán, a 650 metros de altitude, sua extensão é de 541 ha, tem 65 m de profundidade e seus afluentes são os Rios Chavarria, Las Moras, El Chorro e Singuil. O clima é temperado, com invernos secos e verões chuvosos. O objetivo do estudo foi caracterizar as propriedades físicas e químicas das águas limnéticas da represa e da foz dos seus afluentes, para interpretar a qualidade de suas águas. MÉTODOS: Realizaram-se amostragens estacionais de agosto de 2010 até maio de 2012. As variáveis medidas in situ foram: temperatura, transparência, pH, condutividade elétrica, e no laboratório foram analisados: oxigênio dissolvido, demanda bioquímica de oxigênio (DBO5), íons principais, compostos nitrogenados e fosfatados. RESULTADOS: O tipo de água é bicarbonatada-cálcica-sódica, de neutra a alcalina e detectou-se estratificação térmica na primavera e verão. Com exceção da zona limnética: fundo, nos meses mais quentes, as águas estiveram bem oxigenadas. Em novembro de 2011 no rio El Chorro, obteve-se o valor mais baixo de transparência: 0,12 m e o valor mais elevado foi de 4,1 m no rio Singuil no inverno de 2010. Observou-se uma mineralização fraca com condutividades que variaram entre 83 e 218 µS.cm-1. A DBO5 atingiu o registro máximo de 183 mg L-1 na primavera de 2010 no rio Singuil. Os valores mais altos dos compostos nitrogenados detectados foram: 7 mg NO3-.L-1 (zona limnética: fundo, agosto/2010), 0,07 mg NO2-.L-1 (rio Las Moras, maio/2011) e 1,8 mg NH4+.L-1 (rio Chavarría, março/2011). No verão de 2012, o ortofosfato atingiu 0,22 mg.L-1 no fundo da zona limnética. A proporção NT/PT mostrou o fósforo como limitante, e o nitrogênio em raras ocasiões. CONCLUSÕES: O ecossistema analisado foi classificado como hipertrófico considerando NT, PT e transparência. O ACP permitiu uma diferenciação estacional dos sítios, os eixos 1 e 2 separaram as amostras de acordo aos gradientes dos nutrientes, oxigênio dissolvido, DBO5 e temperatura.Fil: Tracanna, Beatriz Concepcion. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo. Instituto de Limnología del Noroeste Argentino; Argentina. Fundacion Miguel Lillo. Direccion de Botanica. Instituto de Ficologia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán; ArgentinaFil: Martínez de Marco, Silvia Nelly. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo. Instituto de Limnología del Noroeste Argentino; Argentina. Fundacion Miguel Lillo. Direccion de Botanica. Instituto de Ficologia; ArgentinaFil: Taboada, María de Los Ángeles. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo. Instituto de Limnología del Noroeste Argentino; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán; ArgentinaFil: Mirande, Virginia. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo. Instituto de Limnología del Noroeste Argentino; Argentina. Fundacion Miguel Lillo. Direccion de Botanica. Instituto de Ficologia; ArgentinaFil: Gultemirian, Maria de Lourdes. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán; ArgentinaFil: Isasmendi, Sara Celina. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo. Instituto de Limnología del Noroeste Argentino; Argentin

Modelling the Contribution of Land Use to Nitrate Yield from a Rural Catchment

The nutrient flow dynamics in rural landscapes are among the basic characteristics of landscape functioning. In this study, the ecohydrological model SWAT (Soil and Water Assessment Tool) was applied in a small rural catchment in northwest (NW) Spain to evaluate the contribution of land use on nitrate losses and to assess the relative importance of different pathways by which nitrate is delivered to the drainage network. The model was first calibrated and validated at a monthly time step. The SWAT model performance was satisfactory (R2 > 0.5; Nash-Sutcliffe efficiency (NSE) > 0.5 and percent bias (PBIAS) < 10%) during both the calibration and validation periods, indicating that SWAT predicted the nitrate discharge accurately. Using the calibrated SWAT model, this study showed that agricultural lands, even though they represent only 30% of the catchment, were main contributor to the nitrate losses accounting for about 77% of the total nitrate yield. The model results also indicated that, irrespective of the land use, groundwater flow is the main pathway for nitrate losses (63%); therefore, appropriate management practices aimed at decreasing nitrate leaching will be key factors in reducing nitrate yield in the study catchment