Dual enzyme responsive and targeted nanocapsules for intracellular delivery of anticancer agents

We report the fabrication of dual enzyme responsive hollow nanocapsules which can be targeted to deliver anticancer agents specifically inside cancer cells. The enzyme responsive elements, integrated in the nanocapsule walls, undergo degradation in the presence of either trypsin or hyaluronidase leading to the release of encapsulated drug molecules. These nanocapsules, which were crosslinked and functionalised with folic acid, showed minimal drug leakage when kept in pH 7.4 PBS buffer, but released the drug molecules at a rapid rate in the presence of either one of the triggering enzymes. Studies on cellular interactions of these nanocapsules revealed that doxorubicin loaded nanocapsules were taken up by cervical cancer cells via folic acid receptor medicated endocytosis. Interestingly the nanocapsules were able to disintegrate inside the cancer cells and release doxorubicin which then migrated into the nucleus to induce cell death. This study indicates that these nanocapsules fabricated from biopolymers can serve as an excellent platform for targeted intracellular drug delivery to cancer cells.

Phenyl carbohydrazone conjugated 2-oxoindoline as a new scaffold that augments the DNA and BSA binding affinity and anti-proliferative activity of a 1,10-phenanthroline based copper(II) complex

A new type of copper(II) complex, CuL(phen)(2)](NO3) (CuIP), where L ((E)-N'-(2-oxoindolin-3-ylidene) benzohydrazide) is a N donor ligand and phen is the N, N-donor heterocyclic 1,10-phenanthroline, has been synthesized. The phenyl carbohydrazone conjugated isatin-based ligand L and CuIP were characterized by elemental analysis, infrared, UV-Vis, H-1 and C-13 NMR and ESI-mass spectral data, as well as single-crystal X-ray diffraction. The interaction of calf thymus DNA (CT DNA) with L and CuIP has been investigated by absorption, fluorescence and viscosity titration methods. The complex CuIP displays better binding affinity than the ligand L. The observed DNA binding constant (K-b = 4.15(+/- 0.18) x 10(5) M-1) and binding site size (s = 0.19), viscosity data together with molecular docking studies of CuIP suggest groove binding and/or a partial intercalative mode of binding to CT DNA. In addition, CuIP shows good binding propensity to the bovine serum albumin (BSA) protein, giving a K-BSA value of 1.25(+/- 0.24) x 10(6) M-1. In addition, the docking studies on DNA and human serum albumin (HSA) CuIP interactions are consistent with the consequence of binding experiments. The in vitro anti-proliferative study establishes the anticancer potency of the CuIP against the human cervical (HeLa) and breast (MCF7) cancer cells; noncancer breast epithelial (MCF10a) cells have also been investigated. CuIP shows better cytotoxicity and sensitivity towards cancer cells over noncancer ones than L under identical conditions, with the appearance of apoptotic bodies. (C) 2014 Elsevier B.V. All rights reserved.

An antioxidant nanozyme that uncovers the cytoprotective potential of vanadia nanowires

Nanomaterials with enzyme-like properties has attracted significant interest, although limited information is available on their biological activities in cells. Here we show that V2O5 nanowires (Vn) functionally mimic the antioxidant enzyme glutathione peroxidase by using cellular glutathione. Although bulk V2O5 is known to be toxic to the cells, the property is altered when converted into a nanomaterial form. The Vn nanozymes readily internalize into mammalian cells of multiple origin (kidney, neuronal, prostate, cervical) and exhibit robust enzyme-like activity by scavenging the reactive oxygen species when challenged against intrinsic and extrinsic oxidative stress. The Vn nanozymes fully restore the redox balance without perturbing the cellular antioxidant defense, thus providing an important cytoprotection for biomolecules against harmful oxidative damage. Based on our findings, we envision that biocompatible Vn nanowires can provide future therapeutic potential to prevent ageing, cardiac disorders and several neurological conditions, including Parkinson's and Alzheimer's disease.

Mesoporous silica-chondroitin sulphate hybrid nanoparticles for targeted and bio-responsive drug delivery

This work proposes the fabrication of a novel targeted drug delivery system based on mesoporous silica-biopolymer hybrids that can release drugs in response to biological stimuli present in cancer cells. The proposed system utilizes mesoporous silica nanoparticles as a carrier to host the drug molecules. A bio-polymer cap is attached onto these particles which serves the multiple functions of drug retention, targeting and bio-responsive drug release. The biopolymer chondroitin sulphate used here is a glycosaminoglycan that can specifically bind to receptors over-expressed in cancer cells. This molecule also possesses the property of disintegrating upon exposure to enzymes over-expressed in cancer cells. When these particles interact with cancer cells, the chondroitin sulphate present on the surface recognizes and attaches onto the CD44 receptors facilitating the uptake of these particles. The phagocytised particles are then exposed to the degradative enzymes, such as hyaluronidase present inside the cancer cells, which degrade the cap resulting in drug release. By utilizing a cervical cancer cell line we have demonstrated the targetability and intracellular delivery of hydrophobic drugs encapsulated in these particles. It was observed that the system was capable of enhancing the anticancer activity of the hydrophobic drug curcumin. Overall, we believe that this system might prove to be a valuable candidate for targeted and bioresponsive drug delivery.

Photocytotoxic luminescent lanthanide complexes of DTPA-bisamide using quinoline as photosensitizer

Lanthanide complexes Ln(DTPAAQ)(DMF)] (1-3) (Ln - Pr (1), Eu (2), Tb (3), H(3)DTPAAQ - N, N `'-bis(3-amidoquinolyl) diethylenetriamine-N, N', N `'-triacetic acid, DMF - N, N-dimethylformamide) were studied for their structures, photophysical properties, DNA and protein binding, DNA photocleavage, photocytotoxicity and cellular internalization. The crystal structures of complexes Ln(DTPAAQ)(DMF)] (1-3) display a discrete mononuclear nine-coordinate {LnN(3)O(6)} tricapped-trigonal prism (TTP) coordination geometry. The europium and terbium complexes show strong luminescence properties in the visible region having a long luminescence lifetime (tau = 0.51-0.64 ms). The conjugated 3-aminoquinoline moieties act as efficient light harvesting antennae, which upon photoexcitation transfer their energy to Eu(III) or Tb(III) for their characteristic D-5(0) -> F-7(J) or D-5(4) -> F-7(J) f-f transitions respectively. The complexes display efficient binding affinity to DNA (K-b = 3.4 x 10(4) - 9.8 x 10(4) M-1) and BSA (KBSA = 3.03 x 10(4) - 6.57 x 10(4) M-1). Europium and terbium complexes give enhanced luminescence upon interacting with CT-DNA suggesting possible luminescence-based sensing applications for these complexes. Complexes 1-3 show moderate cleavage of supercoiled (SC) DNA to its nicked circular (NC) form on exposure to UV-A light of 312 nm involving formation of singlet oxygen (O-1(2)) and hydroxyl radicals (cOH) in type-II and photoredox pathways. Eu(III) and Tb(III) complexes exhibit remarkable photocytotoxicity with human cervical cancer cell line (HeLa) (IC50 = 20.7-28.5 mM) while remaining essentially noncytotoxic up to 150 mM in the dark. Complexes are nontoxic in nature thus suitable for designing cellular imaging agents. Fluorescence microscopy data reveal primarily cytosolic localization of the Eu(III) and Tb(III) complexes in HeLa cells.

Photocytotoxic ternary copper(II) complexes of histamine Schiff base and pyridyl ligands

Ternary copper(Il) complexes of salicylaldehyde-histamine Schiff base (HL) and pyridyl ligands, viz. Cu(bpy)(L)](ClO4) (1) and Cu(dppz)(L)](C104) (2), where bpy is 2,2'-bipyridine (in 1) and dppz is dipyrido3,2-a:2',3'-c]phenazine (in 2), were synthesized, characterized and their DNA binding, photo-activated DNA cleavage activity and photocytotoxicity studied. The 1:1 electrolytic one-electron paramagnetic complexes showed a d-d band near 670 nm in aqueous DMF (1:1 v/v). The crystal structure of complex 1 showed the metal in CuN4O distorted square-pyramidal geometry. Complex 2 intercalatively binds to calf-thymus (ct) DNA with a binding constant (K-b) of similar to 10(5) M-1. It exhibited moderate chemical nuclease activity but excellent DNA photocleavage activity in red light of 647 nm forming (OH)-O-center dot radicals. It showed remarkable photocytotoxicity in human cervical cancer cells (HeLa) giving IC50 of 1.6 mu M in visible light (400-700 nm) with low dark toxicity. The photo-induced cell death is via generation of oxidative stress by reactive oxygen species.

Remarkable photo-induced anticancer activity of vitamin-S6 Schiff base copper(II) complexes of pyridyl bases

Vitamin-B6 (VB6) Schiff base (H2L) copper(II) complexes of pyridyl bases, viz. Cu(bpy)(L)] (1), Cu(phen)(L)] (2) and Cu(dppz)(L)] (3), where bpy is 2,2'-bipyridine, phen is 1,10-phenanthroline and dppz is dipyrido3,2-a:2',3'c]phenazine are synthesized, characterized and their phto-induced anticancer activity studied. The non-electrolytic one electron paramagnetic complexes exhibit a d-d band near 700 nm in DMF. The dppz complex intercalatively binds to calf-thymus DNA with binding constant (K-b) values of similar to 10(6) M-1. This complex exhibits low chemical nuclease activity but excellent DNA photocleavage activity when irradiated with red light of 705 nm forming (OH)-O-center dot radical. It displays remarkable photocytotoxicity in human cervical cancer cells (HeLa) giving IC50 value of 0.9 mu M in visible light (400-700 nm) while being less toxic in darkness (IC50 : 23 mu M). The cellular uptake of the complexes seems to be via VB6 transporting membrane carrier mediated diffusion pathway. Photo-induced cell death follows apoptotic pathway involving photo-generated intracellular reactive oxygen species.

Inventario agroecológico de las especies de Gallina ciega (Phyllophaga spp) en la Región I de Nicaragua y validación de trampas artesanales de luz para el control de adultos en dos localidades del municipio de Estelí, Nicaragua

De mayo a octubre de 1996 se hizo un inventario agroecológico de las especies de gallina ciega (Phyllophaga spp.) en la región T de Nicaragua con el objetivo de determinar la relación entre la diversidad de especies de gallina ciega con precipitación, tamaño y años de uso de la parcela y historial de daño(%) y validación de trampas luz artesanal para control de adultos de gallina ciega en Miraflor y Almaciguera, con el fin de confirmar su efectividad. Se muestrearon 61 localidades en 18 municipios, agrupados por rangos de precipitación en cuatro estratos. La unidad muestra! fué el número de insectos adultos de gallina ciega recolectados por los mismos agricultores en cada finca. Para la identificación se utilizaron las claves desarrolladas por King (1984); Morón (198S); Woodruffy Beck (1989) y Coto (1990). Se realizó un diagoóstico sobre conocimiento de la plaga y el manejo al cultivo por medio de encuestas. Se hizo análisis de correlación simple. La mayor parte de la encuesta fue analizada en términos de porcentajes. Se utilizó la metodología de captura y recaptura de insectos marcados con esmalte para uñas y escarchas de colores diferentes. Los insectos fueron hllerados desde S hasta 21m de distancia de la trampa marcándose grupos de cinco hasta SO insectos. Se consideró un factor correctivo de comparación entre distancias y analisis de frecuencia de recaptura con observaciones cada 15 minutos. Se realizó muestreo de larvas posterior a los ensayos de liberación. La población de gallina ciega en la región T es diviersa. Se reportan 17 especies de los cuales sólo 9 se identificaron: Phyllophaga (Chlaenobia) aequata (Bates), 1888; P. (Phytalus) cometes Bates, 1888; P. (Rorulenta) elenon.' Saylor, 1938; P. (P) lenis Horn, 1887; P.(P) menetriesi Blanchard, 1850; P. (P) obsoleta (Biancbard), 1850; P. (Chlaenobia) tumulosa (Bates), 1888; P. (Rorulenta) vicina Moser, 1918 y P. yucateca (Bates), 1889. Las especies no identificadas fueron distinguidas por números. Las especies 1, 3, 4, 5, 6, 7, 8 y 9 no se identificaron. P. Lenis, P. yucateca, P, vicina, P.elenon.f, P. menetriesi y P. obsoleta son las especies que, en este orden, sobresalen numéricamente. Las cantidades de especímenes por especies de gallina ciega no presentan relación con rangos de precipitación, historial de daño (%), tamaño y años de uso de la aparcela, ni entre ellas mismas. El 60"/o de los agricultores no conoce al adulto de gallina ciega y el 80% no logra diferenciar claramente los sintomas en una planta dañada. No se registraron recapturas de insectos adultos de gallina ciega en las trampas. El análisis cuantitativo no se realizó. La mayor parte de los insectos liberados establecieron vuelo. Muestreos de larvas de gallina ciega indican que sólo en Miraflor habían cantidades de adultos justificables a los ensayos. El promedio de plantas dañadas por gallina ciega según los agricultores es de 15%. Se recomienda impulsar estudios básicos sobre la diversidad de especies de gallina ciega en el país y difundir el conocimiento sobre la biología y manejo de gallina ciega entre los pequeños agricultores. Futuros ensayos con trampas luz deben considerar el aumento de la calidad y la consistencia de la fuente luminosa y establecerlas cerca de las plantas hospederas de adultos.

Patologías reproductivas de la categoría vaca parida del ganado bovino en la comunidad Wasayamba, municipio de Paiwas, Región Autónoma del Atlántico Sur

El presente estudio se realizó con el objetivo de determinar las principales patologías reproductivas en la hembra bovina de la comunidad de Wasayamba, perteneciente al municipio de Paiwas, departamento de Matagalpa, Región Autónoma del Atlántico Sur (RAAS). Los datos generados se analizaron a través de estadística descriptiva, mediante distribución de frecuencias e histogramas. El tamaño requerido de la muestra para este trabajo fue de 1458 hembras en edad reproductiva pertenecientes a 3 7 fincas de la comunidad de Wasayamba. El encaste de .la vacas que más predominó en la zona fue el cruzamiento de 75% Cebú y 25% Pardo Suizo, con un 78% hembras. El encastes de los sementales que brindaron servicio a 297 hembras fue de 75% Cebú y 25% Pardo Suizo, en segundo lugar está el encaste de 75% Pardo Suizo y 25% Cebú, con el l4% hembras servidas y en tercer lugar los sementales con un encaste de 75% Holstein y 25% Cebú, con el 6% hembras servidas. El estado reproductivo que predominó en este estudio fue de hembras vacías, con el 62% de especímenes, en segundo lugar el 12% y 13% hembras con tres meses de gestación, en tercer lugar el 14% de hembras con dos meses de preñez y después los meses de gestación oscilaron entre cuatro hasta los ocho meses. La relación entre la condición corporal y el estado reproductivo de las hembras evaluadas en este estudio se observó que las vacas que poseían una condición corporal entre 2.0-2.5 se encontraron vacías 62% hembras de las 1458 hembras que fueron evaluadas. La patología reproductiva que más afectó a las hembras bovinas de la comunidad de Wasayamba, al 1ro, 2do y 3er parto respectivamente, fue Anestro Postparto con el 44% de hembras afectadas. En segundo lugar la hipofunción Ovárica con el 16% hembras afectadas y en tercer lugar la Atrofia Ovárica con el 13% de las hembras afectadas. Las hembras con encaste 75% Cebú y 25% Pardo Suizo se vieron afectadas con el Anestro Postparto, en segundo lugar las hembras que poseían encaste entre las razas Cebú y Pardo Suizo con Atrofia Ovárica y en tercer lugar las hembras con encaste 75% Cebú y 25% Pardo Suizo, que se les diagnosticó desviación cervical Los productores plantearon que desparasitaban a sus vacas de manera semestral con un 88% y un 6% de ellos lo hacían de forma trimestral y anual respectivamente. Respecto al estado de vacunación de su ganado bovino el 61% de los productores no vacunan, y el 39% de ellos si realizan esta actividad como parte del plan zoosanitario existente en las fincas.

Diário da Constituinte [gravação de vídeo] : [programa n. 011]

O presidente da Assembleia Nacional Constituinte (ANC) Ulysses Guimarães (PSDB-SP) preside a cerimônia em homenagem ao segundo aniversário de morte de Tancredo Neves. Imagem de Risoleta Neves, esposa de Tancredo Neves, em plenário. O Deputado Pimenta da Veiga (PMDB-MG) discursa em nome do PMDB. O Deputado Aécio Neves (PMDB-MG) agradece a homenagem póstuma ao avô. As três Subcomissões da Educação, Cultura e Esportes; da Ciência e da Comunicação; da Família, do Menor e do Idoso que fazem parte da Comissão da Família, Educação e Cultura, Esporte, Ciência e Tecnologia e da Comunicação reuniram-se para tratar de questões como o livro didático, a destinação de verbas governamentais para o ensino público e privado e da educação pré-escolar. O Senador João Calmon (PMDB-ES) acredita numa projeção da educação. A Subcomissão da Família, do Menor e do Idoso recebeu Comba Marques Porto, representante do Conselho dos Direitos da Mulher, que relatou sobre a natureza da sociedade conjugal. A Deputada Rita Camata (PMDB-ES), em entrevista, aborda também a questão da sociedade conjugal. O Ministro Chefe da Casa Civil, Marco Maciel, esteve presente na Subcomissão do Poder Executivo.