Cryoreservation of alginate-fibrin beads involving bone marrow derived mesenchymal stromal cells by vitrification

Application of cell-–biomaterial systems in regenerative medicine can be facilitated by their successful low temperature preservation. Vitrification, which avoids ice crystal formation by amorphous solidification, is an emerging approach to cryopreservation. Developing vitrification strategy, effective cryopreservation of alginate–fibrin beads with porcine mesenchymal stromal cells has been achieved in this study. The cell–biomaterial constructs were pre-cultured for 20 days before cryopreservation, allowing for cell proliferation and construct stabilization. Ethylene glycol (EG) was employed as the basic cryoprotectant for two equilibration solutions. Successful cryopreservation of the constructs was achieved using vitrification solution composed of penetrating (EG MW 62 Da) and non-penetrating (sucrose MW 342 Da) cryoprotectants. Stepwise procedure of introduction to and removal of cryoprotectants was brief; direct plunging into liquid nitrogen was applied. Cell viability, evaluated by combining live/death staining and confocal laser microscopy, was similar for both control and vitrified cells in the beads. No detectable damage of microstructure of cryopreserved beads was found as shown by scanning electron microscopy. Both osteogenically induced control and vitrified cells in the constructs were equally capable of mineral production and deposition. There was no statistically significant difference in metabolic activity and proliferation between both groups during the entire culture period. Our study leads to the conclusion that the developed cryopreservation protocol allowed to maintain the integrity of the beads while preserving the ability of the pig bone marrow derived mesenchymal stromal cells to proliferate and subsequently differentiate; demonstrating that vitrification is a promising approach for cryopreser-vation of “ready-to-use” cell–biomaterial constructs.

Synthesis, Crystal Structure, and Magnetic Properties of a Ferromagnetically Coupled Angular Trinuclear Copper(11) Complex [Cu3(02CMe)4(bpy)s(HzO)](PF)62

The synthesis, X-ray crystal structure, and magnetic properties of an angular trinuclear copper(II) complex [Cu3(O2CMC)4(bpy)3(H2O)](PF6)2 (1), obtained from a reaction of Cu2(O2CMe)4(H2O)2 With 2,2'-bipyridine (bpy) and NH4PF6 in ethanol, are reported. Complex 1 crystallizes in triclinic space group P1BAR with a = 11.529(1) angstrom, b = 12.121(2) angstrom, c = 17.153(2) angstrom, alpha = 82.01(1)-degrees, beta = 79.42(1)-degrees, gamma = 89.62(1)-degrees, and Z = 2. A total of 6928 data with I > 2.5sigma(I) were refined to R = 0.0441 and R(w) = 0.0557. The structure consists of a trinuclear core bridged by four acetate ligands showing different bonding modes. The coordination geometry at each copper is distorted square-pyramidal with a CuN2O2...O chromophore. The Cu...Cu distances are 3.198(1) angstrom, 4.568(1) angstrom, and 6.277(1) angstrom. There are two monoatomic acetate bridges showing Cu-O-Cu angles of 93.1(1) and 97.5(1)-degrees. Magnetic studies in the temperature range 39-297 K show the presence of a strong ferromagnetically coupled dicopper(II) unit (2J = +158 cm-1) and an essentially isolated copper(II) center (2J' = -0.4 cm-1) in 1. The EPR spectra display an axial spectrum giving g(parallel-to) = 2.28 (A(parallel-to) = 160 X 10(-4) cm-1) and g(perpendicular-to) = 2.06 (A(perpendicular-to) = 12 X 10(-4) cm-1) for the normal copper and two intense isotropic signals with g values 2.70 and 1.74 for the strongly coupled copper pair. The structural features of 1 compare well with the first generation models for ascorbate oxidase.

Evaluación de diferentes tratamientos en el control de yemas axilares del tabaco Nicotiana tabacum L. variedad Burley KY-17

En el complejo tabaco, “Hermanos Mendoza” unida de producción estatal (U: P; E) “Juan José Morales” se estableció en la época lluviosas, periodo comprendido del 25 de Julio de 1985 y parte de la Época seca 19 de Enero 1986 un ensayo cuyos tratamientos estudiados fueron todas las combinaciones de tres deshija dores químicos, tres dosis de cada uno y dos diferentes momentos de aplicación, incluyendo un testigo absoluto (sin deshija), y un testigo relativo (deshije manual). Se planteó como objetivos determinar la efectividad de los diferentes tratamientos de las yemas axilares en la planta de tabaco; además identificar la mejor relación beneficio-costo de los tratamientos estudiados. El diseño empleado fue el de bloques completos al azar con cuatros repeticiones. Los productos conocidos como hidrácida maleica en dosis de 2.70 y kg. i.a/ha, y alcoholes grasos en dosis de 6.80Kg. I.a./ha., ejercieron un control excelente sobre las yemas auxiliares de tabaco, arrojado rendimientos similares al resto de los tratamientos, acepto el tratamiento donde se aplicó hidrácida maleica en dosis de 1.35kg.i.a./ha, a los tres dios después del desmote que fue al que arrojo menor rendimiento en peso seca de hojas. Los mayores rendimientos se obtuvieron con pendimatallina en dosis de 0.5kg.i.a./ha., a los cincos días después del desbotone, y con alcoholes grasos en dosis de 8.5 kg. I.a./a a los ocho días después del desbotone Los mayores resultados sobre el rendimiento se obtuvieron con el tratamiento de alcoholes grasos de dosis de 8.5, kg i.a./ha ., aplicado a los ocho días después del desbotone, determinando una producción de 2.1605 kg/P.U y un óptimo económico que corresponde a un retorno en ganancias neta de C$ 5.39,376.62 (quinientos treinta y nueve mil trescientos setenta y seis córdobas con 62/100 por hectárea, equivalente a C$ 1,7426 córdobas de ganancia neta por cada córdobas investido.