Bioengineering of AAV2 Capsid at Specific Serine, Threonine, or Lysine Residues Improves Its Transduction Efficiency in Vitro and in Vivo

We hypothesized that the AAV2 vector is targeted for destruction in the cytoplasm by the host cellular kinase/ubiquitination/proteasomal machinery and that modification of their targets on AAV2 capsid may improve its transduction efficiency. In vitro analysis with pharmacological inhibitors of cellular serine/threonine kinases (protein kinase A, protein kinase C, casein kinase II) showed an increase (20-90%) on AAV2-mediated gene expression. The three-dimensional structure of AAV2 capsid was then analyzed to predict the sites of ubiquitination and phosphorylation. Three phosphodegrons, which are the phosphorylation sites recognized as degradation signals by ubiquitin ligases, were identified. Mutation targets comprising eight serine (S) or seven threonine (T) or nine lysine (K) residues were selected in and around phosphodegrons on the basis of their solvent accessibility, overlap with the receptor binding regions, overlap with interaction interfaces of capsid proteins, and their evolutionary conservation across AAV serotypes. AAV2-EGFP vectors with the wild-type (WT) capsid or mutant capsids (15 S/T -> alanine A] or 9 K -> arginine R] single mutant or 2 double K -> R mutants) were then evaluated in vitro. The transduction efficiencies of 11 S/T -> A and 7 K -> R vectors were significantly higher (similar to 63-90%) than the AAV2-WT vectors (similar to 30-40%). Further, hepatic gene transfer of these mutant vectors in vivo resulted in higher vector copy numbers (up to 4.9-fold) and transgene expression (up to 14-fold) than observed from the AAV2-WT vector. One of the mutant vectors, S489A, generated similar to 8-fold fewer antibodies that could be cross-neutralized by AAV2-WT. This study thus demonstrates the feasibility of the use of these novel AAV2 capsid mutant vectors in hepatic gene therapy.

An Eighteen Serum Cytokine Signature for Discriminating Glioma from Normal Healthy Individuals

Glioblastomas (GBM) are largely incurable as they diffusely infiltrate adjacent brain tissues and are difficult to diagnose at early stages. Biomarkers derived from serum, which can be obtained by minimally invasive procedures, may help in early diagnosis, prognosis and treatment monitoring. To develop a serum cytokine signature, we profiled 48 cytokines in sera derived from normal healthy individuals (n = 26) and different grades of glioma patients (n = 194). We divided the normal and grade IV glioma/GBM serum samples randomly into equal sized training and test sets. In the training set, the Prediction Analysis for Microarrays (PAM) identified a panel of 18 cytokines that could discriminate GBM sera fromnormal sera with maximum accuracy (95.40%) and minimum error (4.60%). The 18-cytokine signature obtained in the training set discriminated GBM sera from normal sera in the test set as well (accuracy 96.55%; error 3.45%). Interestingly, the 18-cytokine signature also differentiated grade II/Diffuse Astrocytoma (DA) and grade III/Anaplastic Astrocytoma (AA) sera from normal sera very efficiently (DA vs. normal-accuracy 96.00%, error 4.00%; AA vs. normal-accuracy 95.83%, error 4.17%). Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis using 18 cytokines resulted in the enrichment of two pathways, cytokine-cytokine receptor interaction and JAK-STAT pathways with high significance. Thus our study identified an 18-cytokine signature for distinguishing glioma sera fromnormal healthy individual sera and also demonstrated the importance of their differential abundance in glioma biology.

Designing Novel Sulphate-based Ceramic Materials as Insertion Host Compounds for Secondary Batteries

Rechargeable batteries have propelled the wireless revolution and automobiles market over the past 25 years. Developing better batteries with improved energy density demands unveiling of new cathode ceramic materials with suitable diffusion channels and open framework structure. In this pursuit of achieving higher energy density, one approach is to realize enhanced redox voltage of insertion of ceramic compounds. This can be accomplished by incorporating highly electronegative anions in the cathode ceramics. Building on this idea, recently various sulphate- based compounds have been reported as high voltage cathode materials. The current article highlights the use of sulphate (SO4) based cathodes to realize the highest ever Fe3+/Fe2+ redox potentials in Li-ion batteries (LiFeSO4F fluorosulphate: 3.9V vs Li/Li+) and Na-ion batteries (Na2Fe2(SO4)(3) polysulphate: 3.8V vs Na/Na+). These sulphate-based cathode ceramic compounds pave way for newer avenues to design better batteries for future applications.

高速滑坡中的热效应分析

从理论和数值分析两方面讨论滑坡滑带内热致孔隙压力及其对高速滑坡的影响.首先给出土体动力平衡方程、热和孔压的产生与耗散方程构成的热-水压-力学模型,然后在该模型基础上讨论滑带(局部化剪切带)产生的条件,对滑带宽度的演化及影响因素进行分析,最后探讨土体变形产生热而导致高孔隙压力及低摩擦系数的问题.结果表明,当热软化效应超过应变硬化效应后,失稳就将发生.失稳后,土体发生剪切局部化变形,在滑坡的滑面内产生剧烈变形.一般情况下,滑带宽度是变化的,其变化对滑带内应变、应变率有很大的影响,从而导致滑带内孔隙压力、温度等的明显变化.滑坡的加速运动是由于剪切带中材料的变形,热被集中于剪切带内,而导致其中孔隙压力的快速上升和土体强度的快速下降引起的.

Efecto de diferentes niveles de aplicación de fertilizante nitrogenado sobre la producción de semilla del Andropogon gayanus kunth var. CIAT 621 (Gamba), en la zona de Carazo

El estudio se desarrollo de junio a diciembre de 1994, en la finca "El Pastor" municipio de Santa Teresa, Departamento de Carazo. Ubicada a una elevación de 367 msnm. y entre las latitudes 1301' y 1302'. La zona registró tamperaturas promedios de 24°C y unos 1550 mm de precipitación media por año; por lo cual es posible considerar la zona agroecológica como trópico seco. Se evaluó el efecto de diferentes niveles de fertilizante nitrogenado sobre la producción de semilla de Andropogon gayanus Kunth, para ello se utilizó un diseño experimental de Bloques Completos al Azar (BCA) y se estudió el efecto de un solo factor (Niveles de fertilizante) con cuatro repeticiones formándose un total de veinte tratamientos 2 El ensayo se realizó en un área total de 875 m en la toma de muestras se empleó el método del metro cuadrado para la evaluación de- MS y forraje verde. En cambio, para evaluar la producción de semilla cruda se evaluó el área útil (3x4 m) de parcela. No se realizó poda de control, a causa del mal invierno. Se procedió a aplicar de una sola vez los diferentes niveles de fertilizante. El estudio estadístico contempló el uso del análisis de la varianza (ANDEVA) y se hizo una separación de medias por la prueba de Duncan. Se midió la influencia porcentual de cinco niveles de fertilizante (0, 25, 50, 75, 100 Kg de Urea, 46% N2/h3.), sobre la producción de semilla cruda de la parcela útil l (SCPU), también el Porcentaje de semilla pura ajustada (PSPA), kilogramos de forraje verde por m 2 (KGFV), porcentaje de materia seca (%MS) y• porcent.aje de viabilidad de la semilla (% VIAB>. Las variables% MS y Semilla cruda evaluadas resultaron con valores significativos para los bloques, a excepción del % SPA, % VIAB y Forraje verde. Para los tratamientos resultó significativ solamente la variable Forraje verde al (P<0.05>. Al evaluar los resultados obtenido sometiendo las variables en estudio al análisis estadístico y considerando el factor costo de producción- rentabilidad y rendimiento productivo en el campo; seleccionamos el tratamiento 50 Kg urea 146%/N,)/ha como el mas idóneo para la producción de semilla del Andropogon gayanus para la zona evaluada.

Evaluación de los anabólicos de crecimiento implix, ganamax, ralgro en novillos (finalización) en el trópico seco de Nicaragua

Con el propósito de evaluar el efecto de tres promotores de crecimiento Implix, Ralqro, Ganamax y un qrupo Testiqo (4 tratamientos), sobre variables de rendimiento en novillos de repasto Criollos x Brahaman con niveles de encaste desconocido, en el trópico seco de Nicaragua (Nandaime) 20 novillos fueron distribuidos al azar, 5 novillos por tratamiento. El estudio tuvo una duración de 3 meses, tiempo en el cual los animales fueron Implantados con los anabólicos antes mencionados. Las variables estudiadas fueron ganancia media diaria (GMD), peso de la canal (PCC), peso a los 90 días (P90d). Los datos obtenidos para determinar estas variables fueron peso inicial, peso a los 15 días, peso final. Los datos obtenidos para determinar estas variables se sometieron a un análisis de varianza y luego a una separación de medios a través de la prueba de DUNCAN. Los tratamientos no tuvieron efecto (P>0.05) sobre la características física de la canal; sin embargo se obtuvo mayor porcentaje de carne maqra y menos grasa en los animales implantados. Los resultados de las variables en estudio presentaron los siguientes valores: Ganamax (T1)tuvo una GMD de 380 grs, peso a los 90 días 392 kq y rendimiento en canal caliente de 194 kq, sequido de Implix (T2l con GMD 360 grs, peso 90 días de 380 kq y rendimiento en canal caliente 189 kg, el tratamiento Ralqro (T3) tuvo un peso a los 90 días de 375 kq obteniendo un peso final igual que el tratamiento Testigo, GMD 260 grs para Ralgro y 300 grs para el tratamiento Testigo, un rendimiento en canal caliente 186 kq para e1 tratamiento Ralqro Y 185 kq para el tratamiento Testigo.

Effect of hydrogen dilution on the deposition of carbon-rich a-SiC:H films by the electron cyclotron resonance method

The deposition of hydrogenated amorphous silicon carbide (a-SiC:H) films from a mixture of silane, acetylene and hydrogen gas using the electron cyclotron resonance chemical vapour deposition (ECR-CVD) process is reported. The variation in the deposition and film characteristics such as the deposition rate, optical band gap and IR absorption as a function of the hydrogen dilution is investigated. The deposition rate increases to a maximum value of about 250 Å min-1 at a hydrogen dilution ratio of about 20 (hydrogen flow (sccm)/acetylene + silane flow (sccm)) and decreases in response to a further increase in the hydrogen dilution. There is no strong dependence of the optical band gap on the hydrogen dilution within the dilution range investigated (10-60) and the optical band gap calculated from the E04 method varied marginally from about 2.85 to 3.17 eV. The room temperature photoluminescence (PL) peak energy and intensity showed a prominent shift to a maximum value of about 2.17 eV corresponding to maximum PL intensity at a moderate hydrogen dilution of about 30. The PL intensity showed a strong dependence on the hydrogen dilution variation.

Influence of Obstacle-Produced Turbulence on Development of Premixed Flames

An investigation into influence of obstructions on premixed flame propagation has been carried out in a semi-open tube. It is found that there exists flame acceleration and rising overpressure along the path of flame due to obstacles. According to the magnitude of flame speeds, the propagation of flame in the tube can be classified into three regimes: the quenching, the choking and the detonation regimes. In premixed flames near the flammability limits, the flame is observed first to accelerate and then to quench itself after propagating past a certain number of obstacles. In the choking regime, the maximum flame speeds are somewhat below the combustion product sound speeds, and insensitive to the blockage ratio. In the more sensitive mixtures, the transition to detonation (DDT) occurs when the equivalence ratio increases. The transition is not observed for the less sensitive mixtures. The dependence of overpressure on blockage ratio is not monotonous. Furthermore, a numerical study of flame acceleration and overpressure with the unsteady compressible flow model is performed, and the agreement between the simulation and measurements is good.

First human hNT neurons patterned on parylene-C/silicon dioxide substrates: Combining an accessible cell line and robust patterning technology for the study of the pathological adult human brain

In this communication, we describe a new method which has enabled the first patterning of human neurons (derived from the human teratocarcinoma cell line (hNT)) on parylene-C/silicon dioxide substrates. We reveal the details of the nanofabrication processes, cell differentiation and culturing protocols necessary to successfully pattern hNT neurons which are each key aspects of this new method. The benefits in patterning human neurons on silicon chip using an accessible cell line and robust patterning technology are of widespread value. Thus, using a combined technology such as this will facilitate the detailed study of the pathological human brain at both the single cell and network level. © 2010 Elsevier B.V.

Micro-systems for Optical Protein-Chip

Protein-Chip as micro-assays for the determination of protein interaction, the analysis, the identification and the purification of proteins has large potential applications. The Optical Protein-Chip is able to detect the multi-interaction of proteins and multi-bio-activities of molecules directly and simultaneously with no labeling. The chip is a small matrix on solid substrate containing multi-micro-area prepared by microfabrication with photolithography or soft lithography for surface patterning, and processed with surface modification which includes the physical, chemical, and bio-chemical modifications, etc. The ligand immobilization, such as protein immobilization, especially the oriented immobilization with low steric hindrance and high bio-specific binding activity between ligand and receptor is used to form a sensing surface. Each area of the pattern is corresponding to only one bioactivity. The interval between the areas is non-bioactive and optically extinctive. The affinity between proteins is used to realize non-labeling microassays for the determination of protein identification and protein interaction. The sampling of the chip is non-disturbing, performed with imaging ellipsometry and image processing on a database of proteins.

Efecto de seis tratamientos nitrogenados bajo riego localizado de 2.81 de agua/metro lineal/día en la producción de chilote en el cultivo del maíz (Zea Mays L.), variedad NB-S, a una densidad poblacional de 125,000 ptas ha-1

El presente trabajo de investigación se llevó a cabo en la época seca que va del 28 de Febrero al 9 de Mayo del año 2012, en los terrenos de la Universidad Nacional Agraria, ubicada en el municipio de Managua kilómetro 12 ½ carretera norte del departamento de Managua, teniendo su ubicación geográfica correspondiente 12º08 ́56.52 ́ ́ latitud norte y 86º09 ́36.02 ́ ́ longitud oeste y a una altura de 56 metros sobre el nivel del mar (m.s.n.m.), presentando temperaturas promedios de 28 ºC, precipitacion es promedios de 950 mm anuales y una humedad relativa promedio de 71 %. El propósito del experimento fue evaluar el efecto de seis tratamientos nitrogenados (A:50 kg ha-1 de Nitrógeno, aplicados a los 21 ddg; B:50 kg ha-1 de Nitrógeno, aplicado el 50 % de la dosis a los 21 ddg y 50 % aplicado a los 41 ddg; C: 50 kg ha-1 de Nitrógeno, aplicado el 100 % de la dosis a los 41 ddg; D: 100 kg ha-1 de Nitrógeno, aplicados a los 21 ddg; E: 100 kg ha-1 de Nitrógeno,aplicados 50 % de la dosis a los 21 ddg y 50 % de la dosis a los 41 ddg; F: 100 kg ha-1 de nitrógeno, aplicado 100 % de la dosis a los 41 ddg) bajo riego localizado (utilizado como herramienta para la mejor disolución de los nutrientes aplicados) de 2.8 litros de agua por metro lineal por día en la producción de chilote en el cultivo del maíz (Zea mays L.), variedad NB-S, a una densidad poblacional de 125 000 ptas ha-1. Aplicando una fertilización base con completo 10-30-10 (194 kg ha-1). El ensayo se estableció con un diseño experimental de bloques completos al azar (BCA), unifactorial, con 6 tratamientos y 4 repeticiones. Las variables evaluadas durante el crecimiento fueron: Altura de planta (cm), diámetro del tallo (cm), número de hojas por plantas y al momento de la cosecha del chilote fueron: Altura de la primera y segunda inserción del chilote, peso del chilote con y sin brácteas (kg), longitud del chilote con y sin brácteas (cm), diámetro del chilote con y sin bráctea en cm, y rendimiento de chilote con bráctea (Kg ha-1).A cada una de las variables evaluadas se sometió a un análisis de varianza (ANDEVA) y separación de medias por la prueba de rangos múltiples de Duncan al 5 % de confiabilidad, el análisis se ejecutó utilizando el software estadístico Infostat. De los 6 tratamientos evaluados, el tratamiento E indujo al mayor rendimiento de chilote, con una producción de 4,029.29 kg ha-1 con un beneficio neto de 16,395.89 C$ ha-1 y una tasa de retorno marginal del 317.26 por ciento.

Thermocapillary Flows In Liquid Bridges Of Molten Tin With Small Aspect Ratios

Numerical simulations were conducted to study thermocapillary flows in short half-zone liquid bridges of molten tin with Prandtl number Pr = 0.009, under ramped temperature difference. The spatio-temporal structures in the thermocapillary flows in short half-zone liquid bridges with aspect ratios As = 0.6, 0.8, and 1.0 were investigated. The first critical Marangoni numbers were compared with those predicted by linear stability analyses (LSA). The second critical Marangoni numbers for As = 0.6 and 0.8 were found to be larger than that for As = 1.0. The time evolutions of the thermocapillary flows exhibited unusual features such as a change in the azimuthal wave number during the three-dimensional stationary (non-oscillating) flow regime, a change in the oscillation mode during the three-dimensional oscillatory flow regime, and the decreasing and then increasing of amplitudes in a single oscillation mode. The effects of the ramping rate of the temperature difference on the flow modes and critical conditions were studied as well. In this paper, the experimental observability of the critical conditions was also discussed. (C) 2008 Elsevier Inc. All rights reserved.