Charge-transfer-driven inclusion of neutral TCNQ molecules in the galleries of a layered double hydroxide: an experimental and computational study

The layered double hydroxides (LDH) or anionic clays are an important class of ion-exchange materials. They consist of positively charged brucite-like inorganic sheets with charge-compensating exchangeable anions in the interlamellar space. Here we show how neutral TCNQ (7,7,8,8-tetracyanoquinodimethane) molecules can be included within the galleries of an LDH. To do so, we exploit the fact that TCNQ is a good electron acceptor that forms donor acceptor complexes with a variety of donors. The electron donor aniline was intercalated into a Mg-Al LDH as p-aminobenzoate (AB) ions by a conventional ion-exchange reaction. We show here that neutral TCNQ molecules may be driven into the galleries of the layered solid by charge-transfer complex formation with the intercalated p-aminobenzoate anions. We use diffraction and spectroscopic measurements in combination with molecular dynamics simulations and quantum chemical calculations to establish the nature of interactions and arrangement of the charge-transfer complex within the galleries of the layered double hydroxide. Electrostatic interactions between the TCNQ molecules and the anchored AB ions, subsequent to charge transfer, are the driving force for the inclusion of TCNQ molecules in the galleries of the LDH.

Cytotoxicity of aluminium oxide nanoparticles towards fresh water algal isolate at low exposure concentrations

The growing commercial applications had brought aluminium oxide nanoparticles under,toxicologists' purview. In the present study, the cytotoxicity of two different sized aluminium oxide nanoparticles (ANP(1), mean hydrodynamic diameter 82.6 +/- 22 nm and ANP(2), mean hydrodynamic diameter 246.9 +/- 39 nm) towards freshwater algal isolate Chlorella ellipsoids at low exposure levels (<= 1 mu g/mL) using sterile lake water as the test medium was assessed. The dissolution of alumina nanoparticles and consequent contribution towards toxicity remained largely unexplored owing to its presumed insoluble nature. Herein, the leached Al3+ ion mediated toxicity has been studied along with direct particulate toxicity to bring out the dynamics of toxicity through colloidal stability, biochemical, spectroscopic and microscopic analyses. The mean hydrodynamic diameter increased with time both for ANP(1) 82.6 +/- 22 nm (0 h) to 246.3 +/- 59 nm (24h), to 1204 +/- 140 nm (72 h)] and ANP(2) 246.9 +/- 39 nm (Oh) to 368.28 +/- 48 nm (24 h), to 1225.96 +/- 186 nm (72 h)] signifying decreased relative abundance of submicron sized particles (<1000 nm). The detailed cytotoxicity assays showed a significant reduction in the viability dependent on dose and exposure. A significant increase in ROS and LDH levels were noted for both ANPs at 1 mu g/mL concentration. The zeta potential and FT-IR analyses suggested surface chemical interaction between nanoparticles and algal cells. The substantial morphological changes and cell wall damage were confirmed through microscopic analyses (SEM, TEM, and CLSM). At 72 h, significant Al3+ ion release in the test medium 0.092 mu g/mL for ANP(1), and 0.19 mu g/mL for ANP(2)] was noted, and the resulting suspension containing leached ions caused significant cytotoxicity, revealing a substantial ionic contribution. This study indicates that both the nano-size and ionic dissolution play a significant role in the cytotoxicity of ANPs towards freshwater algae, and the exposure period largely determines the prevalent mode of nano-toxicity.

Synthesis and evaluation of 2,5-di(4-aryloylaryloxymethyl)-1,3,4-oxadiazoles as anti-cancer agents

A series of 2,5-di(4-aryloylaryloxymethyl)-1,3,4-oxadiazoles 9a-j were obtained via multistep synthesis from hydroxybenzophenones 4a-e. The cytotoxicity of compounds 9a-j was evaluated against human leukemia cell lilies (K562 and CEM). The compounds exhibited moderate to good anti-cancer activity with compounds 9b and 9i having a chloro group exhibiting the best activity (IC50 = 10 mu M). Compound 9i exhibited activity against both the cell lines and 9b only exhibited activity against CEM. Further, a lactate dehydrogenase (LDH) assay and DNA fragmentation studies of the compounds 9a-j were also performed. (C) 2013 Elsevier Masson SAS. All rights reserved.

Vertical electric field stimulated neural cell functionality on porous amorphous carbon electrodes

We demonstrate the efficacy of amorphous macroporous carbon substrates as electrodes to support neuronal cell proliferation and differentiation in electric field mediated culture conditions. The electric field was applied perpendicular to carbon substrate electrode, while growing mouse neuroblastoma (N2a) cells in vitro. The placement of the second electrode outside of the cell culture medium allows the investigation of cell response to electric field without the concurrent complexities of submerged electrodes such as potentially toxic electrode reactions, electro-kinetic flows and charge transfer (electrical current) in the cell medium. The macroporous carbon electrodes are uniquely characterized by a higher specific charge storage capacity (0.2 mC/cm(2)) and low impedance (3.3 k Omega at 1 kHz). The optimal window of electric field stimulation for better cell viability and neurite outgrowth is established. When a uniform or a gradient electric field was applied perpendicular to the amorphous carbon substrate, it was found that the N2a cell viability and neurite length were higher at low electric field strengths (<= 2.5 V/cm) compared to that measured without an applied field (0 V/cm). While the cell viability was assessed by two complementary biochemical assays (MTT and LDH), the differentiation was studied by indirect immunostaining. Overall, the results of the present study unambiguously establish the uniform/gradient vertical electric field based culture protocol to either enhance or to restrict neurite outgrowth respectively at lower or higher field strengths, when neuroblastoma cells are cultured on porous glassy carbon electrodes having a desired combination of electrochemical properties. (C) 2013 Elsevier Ltd. All rights reserved.

Patterned growth and differentiation of neural cells on polymer derived carbon substrates with micro/nano structures in vitro

The growth of neuroblastoma (N2a) and Schwann cells has been explored on polymer derived carbon substrates of varying micro and nanoscale geometries: resorcinol-formaldehyde (RE) gel derived carbon films and electrospun nanofibrous (similar to 200 nm diameter) mat and SU-8 (a negative photoresist) derived carbon micro-patterns. MTT assay and complementary lactate dehydrogenase (LDH) assay established cytocompatibility of RE derived carbon films and fibers over a period of 6 days in culture. The role of length scale of surface patterns in eliciting lineage-specific adaptive response along, across and on the interspacing between adjacent micropatterns (i.e., ``on'', ``across'' and ``off'') has been assayed. Textural features were found to affect 3',5'-cyclic AMP sodium salt-induced neurite outgrowth, over a wide range of length scales: from similar to 200 nm (carbon fibers) to similar to 60 mu m (carbon patterns). Despite their innate randomness, carbon nanofibers promoted preferential differentiation of N2a cells into neuronal lineage, similar to ordered micro-patterns. Our results, for the first time, conclusively demonstrate the potential of RE-gel and SU-8 derived carbon substrates as nerve tissue engineering platforms for guided proliferation and differentiation of neural cells in vitro. (C) 2013 Elsevier Ltd. All rights reserved.

Metallacyclocumulenes: A Theoretical Perspective on the Structure, Bonding, and Reactivity

CONSPECTUS: Transition metals help to stabilize highly strained organic fragments. Metallacycles, especially unsaturated ones, provide much variety in this area. We had a sustained interest in understanding new C-C bond formation reactions affected by binuclear transition metal fragments Cp2M. One such study led to the exploration of the bimetallic C-C cleavage and coupled complexes, where the acetylide ligands bridge two metal atoms. The underlying M-C interaction in these complexes inspired the synthesis of a five-membered cyclocumulene complex, which opened a new phase in organometallic chemistry. The metallacyclocumulene produces a variety of C-C cleavage and coupled products including a radialene complex. Group 4 metallocenes have thus unlocked a fascinating chemistry by stabilizing strained unsaturated C4 organic fragments in the form of five-membered metallacyclocumulenes, metallacyclopentynes, and metallacycloallenes. Over the years, we have carried out a comprehensive theoretical study to understand the unusual stability and reactivity of these metallacycles. The unique (M-C-beta) interaction of the internal carbon atoms with the metal atom is the reason for unusual stability of the metallacycles. We have also shown that there is a definite dependence of the C-C coupling and cleavage reactions on the metal of metallacyclocumulenes. It demonstrates unexpected reaction pathways for these reactions. Based on this understanding, we have predicted and unraveled the stabilization factors of an unusual four-membered metallacycloallene complex. Indeed, our prediction about a four-membered heterometallacycle has led to an interesting bonding situation, which is experimentally realized. This type of M-C bonding is intriguing from a fundamental perspective and has great relevance in synthesizing unusual structures with interesting properties. In this Account, we first give a short prologue of what led to the present study and describe the salient features of the structure and bonding of the metallacyclocumulenes. The unusual reaction pathway of this metallacycle is explored next. Similar features of the metallacyclopentynes and metallacycloallenes are briefly mentioned. Then, we discuss the exploitation of the unique M-C bonding to design some exotic molecules such as a four-membered metallacycloallene complex. Our efforts to build a conceptual framework to understand these metallacycles and to exploit their chemistry continue.

Purification of a recombinant histidine-tagged lactate dehydrogenase from the malaria parasite, Plasmodium vivax, and characterization of its properties

Lactate dehydrogenase (LDH) of the malaria parasite, Plasmodium vivax (Pv), serves as a drug target and immunodiagnostic marker. The LDH cDNA generated from total RNA of a clinical isolate of the parasite was cloned into pRSETA plasmid. Recombinant his-tagged PvLDH was over-expressed in E. coli Rosetta2DE3pLysS and purified using Ni2+-NTA resin giving a yield of 25-30 mg/litre bacterial culture. The recombinant protein was enzymatically active and its catalytic efficiency for pyruvate was 5.4 x 10(8) min(-1) M-1, 14.5 fold higher than a low yield preparation reported earlier to obtain PvLDH crystal structure. The enzyme activity was inhibited by gossypol and sodium oxamate. The recombinant PvLDH was reactive in lateral flow immunochromatographic assays detecting pan- and vivax-specific LDH. The soluble recombinant PvLDH purified using heterologous expression system can facilitate the generation of vivax LDH-specific monoclonals and the screening of chemical compound libraries for PvLDH inhibitors.

Association Rule Sharing Model for Privacy Preservation and Collaborative Data Mining Efficiency

The disclosure of information and its misuse in Privacy Preserving Data Mining (PPDM) systems is a concern to the parties involved. In PPDM systems data is available amongst multiple parties collaborating to achieve cumulative mining accuracy. The vertically partitioned data available with the parties involved cannot provide accurate mining results when compared to the collaborative mining results. To overcome the privacy issue in data disclosure this paper describes a Key Distribution-Less Privacy Preserving Data Mining (KDLPPDM) system in which the publication of local association rules generated by the parties is published. The association rules are securely combined to form the combined rule set using the Commutative RSA algorithm. The combined rule sets established are used to classify or mine the data. The results discussed in this paper compare the accuracy of the rules generated using the C4. 5 based KDLPPDM system and the CS. 0 based KDLPPDM system using receiver operating characteristics curves (ROC).

DNA intercalative 4-butylaminopyrimido4',5':4,5]thieno(2,3-b)quinoline induces cell cycle arrest and apoptosis in leukemia cells

DNA intercalators are one of the interesting groups in cancer chemotherapy. The development of novel anticancer small molecule has gained remarkable interest over the last decade. In this study, we synthesized and investigated the ability of a tetracyclic-condensed quinoline compound, 4-butylaminopyrimido4',5':4,5]thieno(2,3-b)quinoline (BPTQ), to interact with double-stranded DNA and inhibit cancer cell proliferation. Circular dichroism, topological studies, molecular docking, absorbance, and fluorescence spectral titrations were employed to study the interaction of BPTQ with DNA. Cytotoxicity was studied by performing 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assay. Further, cell cycle analysis by flow cytometry, annexin V staining, mitochondrial membrane potential assay, DNA fragmentation, and western blot analysis were used to elucidate the mechanism of action of BPTQ at the cellular level. Spectral, topological, and docking studies confirmed that BPTQ is a typical intercalator of DNA. BPTQ induces dose-dependent inhibitory effect on the proliferation of cancer cells by arresting cells at S and G2/M phase. Further, BPTQ activates the mitochondria-mediated apoptosis pathway, as explicated by a decrease in mitochondrial membrane potential, increase in the Bax:Bcl-2 ratio, and activation of caspases. These results confirmed that BPTQ is a DNA intercalative anticancer molecule, which could aid in the development of future cancer therapeutic agents.

Synthesis and evaluation of 2,5-di(4-aryloylaryloxymethyl)-1,3,4-oxadiazoles as anti-cancer agents

A series of 2,5-di(4-aryloylaryloxymethyl)-1,3,4-oxadiazoles 9a-j were obtained via multistep synthesis from hydroxybenzophenones 4a-e. The cytotoxicity of compounds 9a-j was evaluated against human leukemia cell lilies (K562 and CEM). The compounds exhibited moderate to good anti-cancer activity with compounds 9b and 9i having a chloro group exhibiting the best activity (IC50 = 10 mu M). Compound 9i exhibited activity against both the cell lines and 9b only exhibited activity against CEM. Further, a lactate dehydrogenase (LDH) assay and DNA fragmentation studies of the compounds 9a-j were also performed. (C) 2013 Elsevier Masson SAS. All rights reserved.

In Vitro Production of Echioidinin, 7-O-Methywogonin from Callus Cultures of Andrographis lineata and Their Cytotoxicity on Cancer Cells

Andrographis lineata is an herbal medicinal plant used in traditional medicine as a substitute for Andrographis paniculata. Here, using mature leaf explants of A. lineata we demonstrate for the first time the callus induction established on MS medium containing 1.0 mg l(-1) IAA. Dried callus was subjected to solvent extraction with acetone. Further the acetone residue was separated by silica gel column chromatography, crystallized and characterized on the basis of nuclear magnetic resonance (proton and c13) and liquid chromatographic mass spectroscopy. This analysis revealed the occurrence of two known flavones namely, 7-O-methylwogonin (MW) and Echioidinin (ED). Furthermore, these compounds were tested for their cytotoxicity against leukemic cell line, CEM. We identify that ED and MW induced cytotoxicity in a time-and concentration-dependent manner. Further increase in the LDH release upon treatment with ED and MW further confirmed our cytotoxicity results against leukemic cell line. Strikingly, MW was more potent than ED when compared by trypan blue and MTT assays. Our results recapitulate the utility of callus cultures for the production of plant specific bioactive secondary metabolites instead of using wild plants. Together, our in vitro studies provide new insights of A. lineata callus cultures serving as a source for cancer chemotherapeutic agents.

Prevalencia de Cysticercus bovis en el matadero de Amerrisque

En el presente trabajo se analizó la situación existente en relación a la Prevalencia de Cysticercosis bovina en la Empresa Agrícola Centro Americana, S.A. Matadero de Amerrisque, ubicado en el km 128 carretera al Rama, Juigalpa, Chontales. Esta empresa se fundó en el año 1977 y del total de su producción el 98% va destinado a la exportación y el 2% para consumo local. Los análisis se realizaron basándose en datos de la Inspección Post-morten de los años 1982, 1984, 1990, 1991, 1992, para loe cuales se determinó el Año de mayor prevalencia, Categoría más afectada, Departamento de mayor prevalencia y Pérdidas económicas. Para la valoración estadística de los datos registrados fue utilizado el Método de Mínimos Cuadrados Generalizado, usando el procedimiento CATHOD del Paquete Statical Analisis System. Loa resultados obtenidos en el Análisis de Varianza, demuestran que existen diferencias significativas entre años, con un nivel de significancia de (P < 0.05), obteniéndose 0.93% de infestación promedio en los cinco años evaluados. A su vez también se encontraron diferencias significativas para las categorías (P < 0.05) encontrándose la categoría Buey (C4) la más infestada. Resultando Chontales el departamento con mayor número de animales parasitados, seguido de Boaco, Zelaya y Matagalpa. Las pérdidas económicas para estos años resultaron ser de US$ 71,905.85 para el productor, sumándose a ello, las divisas no captadas con un monto aproximado de US$ 535,511.63 al prescindir de su exportación.

Evaluación de la eficiencia de implantación de especies forrajeras megatérmicas en bajos alcalino-sódicos en la Cuenca del Salado

Resumen: La región denominada Cuenca del Salado es considerada la más importante en la cría vacuna del país, aunque su productividad es muy baja. La mayor existencia de animales, sumado al aumento de la superficie dedicada a la agricultura, ha provocado un aumento de carga animal sobre los pastizales naturales, los cuales constituyen la base forrajera de la actividad ganadera. Surge así, la necesidad de encontrar alternativas tecnológicas que permitan mejorar los niveles productivos y sostener el aumento de carga animal en zonas donde no es viable la agricultura. Frente a esta problemática, se evaluó la posibilidad de mejorar estos ambientes mediante la implantación de especies forrajeras megatérmicas o carbono 4 (C4), las cuales podrían aumentar en forma significativa la receptividad y generar un gran impacto en los balances forrajeros de los sistemas ganaderos de cría. Las especies evaluadas fueron Grama Rhodes Fine Cut, Grama Rhodes Top Cut y Panicum Coloratum Kleinpanic. Dichas gramíneas tienen características muy deseables para las zonas donde se realizaron los ensayos, como ser la tolerancia a la salinidad y al encharcamiento temporal, buena calidad forrajera durante su ciclo de producción, alta producción de materia seca y posibilidad de aprovechamiento a partir del primer año. El ensayo se realizó con el objetivo de determinar la eficiencia de implantación de especies forrajeras megatérmicas o C4, cuantificando las plantas logradas por metro cuadrado. Las evaluaciones se realizaron en cinco establecimientos de la Cuenca del Salado, abarcando diferentes situaciones. Los resultados obtenidos arrojaron diferencias significativas a favor de Grama Rhodes Fine Cut (21 plantas/m2) por sobre la especie Panicum Coloratum Kleinpanic (10 plantas/m2), no pudiéndose determinar diferencias significativas entre Grama Rhodes Top Cut (17,4plantas/m2) y las variedades anteriormente mencionadas.

Evaluación de la producción de biomasa de dos cultivares de Panicum virgatum (L.) durante el período de implantación y el primer año de producción

La Argentina vive, desde hace unos años, un claro desplazamiento de la ganadería hacia zonas marginales, en las cuales imperan condiciones edáficas y climáticas que son generadoras de estrés para una amplia cantidad de forrajeras utilizadas. El objetivo del presente estudio fue evaluar el rendimiento de Panicum virgatum (L.). Esta especie forrajera de metabolismo C4 fue descrita como tolerante a condiciones marginales de crecimiento. Fueron evaluados los cvs. Shawnee y Trailblazer. El experimento se condujo en dos ambientes: I) en el predio perteneciente a la Facultad de Ciencias Agrarias de la Universidad Católica Argentina (Lat.34°34´S, 58°26´O) en el cual se evaluó el rendimiento de ambas variedades sembradas a una distancia de 17,5 cm entre hileras y II) en Luján (Buenos Aires, Lat.34°32´S, 59°16´O) en un suelo Argiudol típico, evaluando solamente al cv. Shawnee a dos distintos distanciamientos entre hileras (17,5 y 35 cm). Ambos sitios experimentales se prepararon según un sistema de labranza convencional. Las siembras se efectuaron los días 3 y 5 de octubre de 2008 (ambiente 1 y 2 respectivamente) en forma manual y con un distanciamiento dentro de la hilera de 5cm y una profundidad de 1cm. El arreglo experimental consistió en un DCA con tres repeticiones. Los datos obtenidos se analizaron mediante un Análisis de la Varianza. Se tomó como criterio de defoliación una altura del follaje de 60cm y dejando una altura remanente de 15cm luego del corte En el ambiente 1, el rendimiento acumulado no difirió estadísticamente entre los genotipos estudiados en las dos campañas evaluadas. Durante el primer año, la producción (expresada en kg MS ha-1 ± EEM) del cv. Shawnee fue de 6.293,33 ± 673,42 mientras que el cv. Trailblazer produjo 5.072,38 ± 1.011,85. En cambio, durante el segundo año se observó un marcado incremento en la producción acumulada registrándose valores de 10.590,48 ± 638,31 y 8.857,14 ± 2187,41 para los cv Trailblazer y Shawnee respectivamente. En el ambiente 2, no se observaron diferencias en el rendimiento del cultivo sembrado a 17,5cm o 35 cm de distancia entre hileras. En efecto, con una separación de 17.5 cm se registró una producción de 8.302,85 ± 342,92 y 26.095,24 ± 3780,99 (año 1 y 2 respectivamente) mientras que a 35 cm se observaron 6.054,28± 1.216,01 y 29.428,57± 5.735,67 (año 1 y 2 respectivamente).Se observó la formación de estructuras reproductivas en ambos genotipos demostrando la adaptabilidad de esta especie a nuestra latitud. Los resultados presentados señalan la utilidad de continuar la evaluación de esta especie incluyendo su evaluación en ambientes donde la misma sea capaz de expresar la tolerancia a los factores abióticos generadores de estrés y su respuesta a otras variables de manejo agronómico.

Estudio de los recursos forrajeros, bajo el dosel de árboles en potreros, estudio de caso, finca Las Mercedes, Managua, Nicaragua

Se condujo un experimento de campo en la finca Las Mercedes, Universidad Nacional Agraria. El objetivo del estudio fue determinar el número de componentes forrajeros herbáceos y su tipo en un sistema de producción pecuario bajo condiciones con y sin árboles. Muestreos iniciales indicaron que en el área experimental existían siete tipos de asociaciones. En base a dichas asociaciones, se dividió el área en siete parcelas. Se determino la composición botánica de la base de los árboles a 10, 20 y más de 20 m., con el objeto de identificar las especies gramíneas y leguminosas presentes en cada distancia. Las variables evaluadas fueron composición botánica del estrato herbáceo, producción de biomasa fresca y seca y altura y cobertura del estrato herbáceo. Se cálculo el área de la copa de los árboles y la altura del fuste de los mismos, con el fin de determinar la relación existente entre éstos con la composición del estrato herbáceo. Se logro determinar que la composición botánica de las plantas herbáceas no varia de un sistema de producción de pastos con y sin árboles. En el ámbito de las distancias, a mayor distancia de la base del árbol el porcentaje de plantas C4 (gramineas forrajeras) aumentan y el porcentaje de plantas C3 (hojas anchas) disminuye. Las plantas C4 (gramineas forrajeras) aumentan su producción de biomasa fresca y seca a medida que se ubican a mayor · distancia de la base de los árboles. Se determino que la altura del fuste y área de la copa de los árboles afectan negativamente la producción de biomasa fresca y seca de la pastura a base de gramineas (C4), así como la morfo-estructura de ésta. En este tipo de sistemas pastoriles se recomienda ubicar las plantas C4 a mayor distancia de la base de los árboles, y las plantas C3, las cuales pueden ser leguminosas forrajeras, a menor distancia.