Magselectofection: A novel integrated technology of magnetic separation and genetic modification of target cells

Research applications and cell therapies involving genetically modified cells require reliable, standardized and cost-effective methods for cell manipulation. The goal of this work is to provide a novel methodology that produces, in a single standardized techonology, genetic modification and cell isolation. We have named this novel procedure ―Magselectofection”. The approach is based on magnetic cell separation and magnetically-guided gene delivery (magnetofection). Optimized gene vectors associated with novel magnetic nanoparticles were formulated to transfect/transduce target cells while they are passaged and separated through a high gradient magnetic field cell separation column. Magnetofection of the Jurkat T cells using selected vector formulations resulted in a significant (up to 4.5-fold) enhancement in both luciferase reporter gene expression and the percentage of cells expressing eGFP, as compared to lipofection. A procedure for vector loading on LS Miltenyi columns was developed that enables up to 100% retention for both non-viral and viral magnetic complexes. We demonstrate, using a model cell mixture of K562 and Jurkat T cells, that the integrated method is highly efficient and specific for the target cell population. This was not only true for the model Jurkat/K562 mixture, but also for Sca-1+ mouse hematopoietic stem cells. With human umbilical cord mesenchymal stem cells (hUC-MSCs), we achieve up to 30% transfected cells with non-viral vector doses as low as 8 pg plasmid DNA per cell and up to 100% transduced cells with a multiplicity of infection of 0.5 TU/cell using lentivirus. Similarly, we obtain 22% eGFP-positive human cord blood hematopoietic stem cells (hCB-HSCs) upon lentiviral magselectofection compared to 0.15% eGFP-positive cells post-standard infection. We achieve up to 50% transduced Sca-1+ mouse stem cells at a lentiviral MOI of 1-3. Up to 5-15% and 20% genetic modified PBMC were found using non-viral and viral magselectofection, respectively. After genetic modification using magselectofection differentiation potential of hCB-HSCs and hUC-MSCs was maintained. Magselectofection requires a minimal number of manipulation steps and results in efficient and specific gene delivery to target cells. This minimizes the necessary vector material while maintaining the cellular differentiation potential of modified stem cells. Magselectofection may become a useful tool for nucleic acid therapy approaches involving ex-vivo genetically modified cells

Análisis de precios de productos básicos en tiendas de autoservicio de Pachuca y el Comercio de Zacualtipán, Hidalgo, 2007-2008

Esta investigación se realizó en tiendas de autoservicio de Pachuca y el comercio local de Zacualtipán, Estado de Hidalgo. Se registraron precios de 24 productos: arroz, tortilla, jitomate, limón, chile jalapeño, leche, huevo, jamón, salchicha, carne de res, carne de pollo, papel sanitario, jabón de polvo, jabón de baño, pasta de dientes, frijol, cebolla, papa, aguacate, naranja, plátano, aceite, sal y azúcar. Las fechas de consulta de precios fueron: 26 de julio de 2007, 25 de enero y 29 de mayo de 2008. La importancia del estudio es el análisis comparativo y evolución de precios. Las tiendas de autoservicio ofrecen 9.7% más barato en el 62.5% de los productos. El comercio local ofrece el 12.7% más barato que las tiendas de autoservicio en frijol, cebolla, papa, aguacate, plátano, aceite, sal y azúcar. Durante el 26 de julio de 2007 al 29 de mayo de 2008, los precios aumentaron 19.8% y 19.6% en el comercio local y tiendas de autoservicio, respectivamente. Palabras clave: Precios, productos básicos, pequeñas y medianas empresas, tiendas de autoservicio, análisis comparativo, comercio

ANÃLISIS DE LA RENTABILIDAD ECONÓMICA DE LA PRODUCCIÓN DE JITOMATE BAJO INVERNADERO EN ACAXOCHITLÃN, HIDALGO

In this study there was evaluated the economic profitability of tomato production under greenhouse conditions four production projects, in an area of 9 040 m2., in the communities of San Martin, San Pedro and Los Reyes Acaxochitlán's municipality, Hidalgo State, during the 2008-2009 cycle. How indicator was used cost/benefit ratio (C/B). Acaxochitlán's municipality produces 1,582.3 tons of tomato in a surface of 65 200 m2. In the four projects reviewed obtained a production of 218 tons, the average selling cosprice was $ 8.25 per kilogram, and sold in the cities of Tulancingo, Acaxochitlán and Pachuca. Tomato production proved a profitable activity in the study region since the projects 1, 2, 3 and 4 showed a ratio C/B of 2.30, 2.65, 3.09 and 1.57, respectively.Economic profitability, cost/benefit ratio, tomato production, marketing., Agribusiness,

DNA Damage Induces p53-dependent Down-regulation of hCHK1

Abstract The levels of the human checkpoint gene hCHK1 were measured in human cancer cells growing in vitro after treatment with the DNA damaging agent cis-dichlorodiammine platinum(II) (DDP). Treatment of human cancer cell lines with DDP induced a decrease in the hCHK1 protein levels starting 6 h after treatment, with a further decline at 24 and 48 h. A similar decrease in the levels of hCHK1 was found at the mRNA level by using Northern blot analysis. By using isogenic cell systems in which p53 was disrupted either by transfection with HPV-E6 or by targeted homologous recombination, we found that the DNA damage-induced down-regulation of hCHK1 was only observable in wild type p53-expressing cells, with only a minor decline in the hCHK1 levels observable 48 h after treatment in cells with disrupted p53. Similarly, treatment of mutant p53-expressing human cancer cell lines with DDP did not result in changes in the levels of hCHK1. The p53-dependent down-regulation of hCHK1 is likely to be at transcriptional levels, as suggested by the lack of down-regulation of the hCHK1 when transfected under the control of a heterologous viral promoter. In addition, p53 is able to down-regulate the luciferase activity under the control of the 5′ flanking region of the hCHK1 gene. The data suggest a strict link between p53 and hCHK1 governing the activation and repression of the G2 checkpoint in which both proteins participate

Glycogen Synthase Kinase 3 (GSK-3) involvement in regulation of mouse embryonic stem cell fate

EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Functionalization of Silica SBA-15 with 3-(2-Aminoethylamino)Propyl Trimethoxysilane in Supercritical CO2 Modified with Methanol or Ethanol for Carbon Capture

The CO2 adsorption process using amine-grafted silica is a promising technology for reducing the CO2 emissions from the power and industry sectors. In this work, silica SBA-15 was functionalized using 3-(2-aminoethylamino)propyl trimethoxysilane (AEAPTS) in supercritical CO2 (scCO2) modified with 10% mol methanol or ethanol. The functionalization experiments were carried out at 323 K and 12.5 MPa, and with reaction times of 2 and 3 h. The molar fraction of AEAPTS in scCO2 plus 10% mol alcohol ranged from 0.5 × 10−3 to 1.8 × 10−3. It was found that as the molar fraction of AEAPTS increased, the amino-grafting density steadily rose, and the pore volume, surface area and pore size of the functionalized silica SBA-15 also decreased gradually. The scCO2 functionalization method was compared to the traditional toluene method. The diamine-SBA-15 prepared in the scCO2 process shows a slightly lower amine-grafting density but a higher surface area and pore volume than the ones obtained using the traditional method. Finally, the excess CO2 adsorption capacity of the materials at different temperatures and low pressure was measured. The diamine-silica SBA-15 displayed moderate excess CO2 adsorption capacities, 0.7–0.9 mmol∙g−1, but higher amine efficiency, ca. 0.4, at 298 K, due to the chemisorption of CO2. These findings show that diamine-grafted silica for post-combustion capture or direct air capture can be obtained using a media more sustainable than organic solvents

Managing with (fluorescent)-biomimetic dendrimers

Dendrimers that are modified through their peripheral groups have been employed for many biomedical purposes. Diagnosis via specific interactions with target proteins is one of the objectives that has been given more attention. In this way, we have been developing different methodologies for the detection of immunoglobulins, as an in vitro method for the diagnosis of allergic reactions to drugs [1], [2]. Specific cell interactions of surfaces grafted dendrimers have been applied to the study of cell adhesion. We have studied also the application of dendrimer-based uneven nanopatterns to evaluate the local RGD surface density effects on cell adhesion, as bioactive substrates to evaluate the impact of the RGD local surface density on the chondrogenic induction of adult human mesenchymal stem cells [3], [4], [5]. Although the use of PAMAM dendrimer has allowed us to develop useful methods for this purpose, for certain objectives we have required the development of new dendrimeric models. We have developed a new approach for the production of allaliphatic polyamide dendrimers (BAPAD) by iterative 3,3′-diaminopivalic acid connections as building blocks for dendrimer construction. These dendrimers were studied in explicit solvent by atomistic forcefield-based molecular dynamics to characterize structural properties such as shape, radius and monomer distribution [6]. Fluorescent labeling of these biomimetic dendrimers has been used as a tool to study their interaction with cells.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

ddRAD-seq para el análisis de asociación genómico de los principales caracteres agronómicos y calidad organoléptica en melocotonero

PósterDouble-digest RAD sequencing (ddRAD-Seq) is a powerful method for SNP discovery at a genome-wide scale. It relies on breaking the genome into a certain size of DNA fragments using two restriction enzymes: i) one common cutter with a short recognition site and ii) a low frequency cutter having a large recognition motif . Amplification step in library preparation can introduce PCR artefacts. Those are expected to skew allele frequency by increasing homozygosity leading to false genotype calls. In this context, we carried out a comparison study between DNA-variants generated with duplicates and those generated after removing with either "'SAMtools:rmdup" or "Stacks clone filter". On the other hand, the accuracy of genetic variants identification is a crucial step towards understanding phenotypical traits and monitoring breeding programs. Thereby, a good combination of computational tools for alignment and variant calling is crucial to tackle the possible artifacts. In response to this challenge, three variant callers (BCFtools, Freebayes and GATK-HaplotypeCaller) were combined on top of the BWA-mem read mapper. SNPs derived from the intersection of these callers were used for a genome wide association study to genetic variants associated with agronomic and organoleptic traits (In this poster, we illustrate only one study case trait). A diverse set of 90 peach accessions were sampled from the Experimental Station of Aula Dei (CSIC) located at Zaragoza (northern of Spain). This germplasm collection includes landraces and modem breeding lines from different origins.EEA San PedroFil: Ksouri, Najla. Consejo Superior de Investigaciones Científicas (CSIC). Estación Experimental Aula Dei; EspañaFil: Sánchez, Gerardo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria San Pedro; ArgentinaFil: Contreras-Moreira, Bruno. Consejo Superior de Investigaciones Científicas (CSIC). Estación Experimental Aula Dei; EspañaFil: Gogorcena Aoiz, Yolanda. Consejo Superior de Investigaciones Científicas (CSIC). Estación Experimental Aula Dei; Españ

Análisis crítico léxico semántico y estadístico de tiempo de silencio de Luis Martín Santos

Los puntos mas relevantes que se que se desarrollaron en este trabajo investigativo fueron los siguientes: los aspectos generales de la investigación que sirvieron para exponer los pasos que debe tener presente el investigador o la investigadora para elegir un tema. En el marco teórico se enfocó los problemas sociales de la época a través de la novela. Se analizó la obra desde el punto de vista léxico-semántico y estadístico, presentado de acuerdo con el orden en que aparecen. De igual modo, a través del marco teórico se expuso los métodos y las técnicas de enseñanza utilizados por los docentes de Licenciatura en Español y los distintos procedimientos de análisis empleados para la correcta interpretación de la investigación. En cuanto al análisis e interpretación de los resultados, se aplicó la encuesta y entrevista a los estudiantes y docentes utilizando diversos instrumentos para comprobar los hechos. También se presentó la propuesta educativa para el desarrollo de reforzamiento y orientación académica. Igualmente se estudió, criticó y valoró la obra, terminando con las conclusiones, recomendaciones y material bibliográfico del trabajo

Implementación de un dashboard basado en indicadores productivos y de comercio exterior para el análisis económico del subsector agrícola en la dirección de estudios económicos, del Ministerio de Desarrollo Agrario y Riego

El presente trabajo se desarrolló basado en los requerimientos de los especialistas de la dirección de Estudios Económicos, la cual requería de información estadística agraria mensual para cumplir con sus objetivos, traduciéndose en productos como informes técnicos, estudios, boletines y notas técnicas, mediante ellos se generaban las notas de prensa correspondientes con el visto bueno de la dirección, como consecuencia de estos trabajos, era sentida la necesidad de contar con una base de datos que sea, recopilada, procesada y puesta al servicio de los analistas, para el estudio e investigación programados en el POI. De allí que el presente trabajo pone a consideración el desarrollo de un dashboard para el análisis económico sectorial en power bi, en el cual se procesó los cuadros estadísticos en formato Excel de las cuatro variables productivas (cosecha, producción, rendimiento y precio en chacra) para luego trabajar la elaboración del diseño de dashboard, El trabajo se realizó en cinco capítulos. El capítulo I introducción, detalla la información de la empresa, reseña histórica, visión, misión, dirección, objetivos y organigrama. Capítulo II marco teórico, capitulo III descripción de la experiencia. capítulo IV resultados y Capítulo V conclusiones y recomendaciones