Tissue engineering of stratified articular cartilage from chondrocyte subpopulations

Objective: To test if subpopulations of chondrocytes from different cartilage zones could be used to engineer cartilage constructs with features of normal stratification. Design: Chondrocytes from the superficial and middle zones of immature bovine cartilage were cultured in alginate, released, and seeded either separately or sequentially to form cartilage constructs. Constructs were cultured for 1 or 2 weeks and were assessed for growth, compressive properties, and deposition, and localization of matrix molecules and superficial zone protein (SZP). Results: The cartilaginous constructs formed from superficial zone chondrocytes exhibited less matrix growth and lower compressive properties than constructs from middle zone chondrocytes, with the stratified superficial-middle constructs exhibiting intermediate properties. Expression of SZP was highest at the construct surfaces, with the localization of SZP in superficial-middle constructs being concentrated at the superficial surface. Conclusions: Manipulation of subpopulations of chondrocytes can be useful in engineering cartilage tissue with a biomimetic approach, and in fabricating constructs that exhibit stratified features of normal articular cartilage. (C) 2003 OsteoArthritis Research Society International. Published by Elsevier Ltd. All rights reserved.

Simulation of RNA silencing pathway for time-dependent transgene transcription rate

The synthesis of dsRNA is analyzed using a pathway model with amplifications caused by the aberrant RNAs. The transgene influx rate is assumed time-decaying considering the fact that the number of transgenes can not be infinite. The dynamics of the transgene induced RNA silencing is investigated using a system of coupled nonautonomous ordinary nonlinear differential equations which describe the model phenomenologically. The silencing phenomena are detected after a period of transcription. Important contributions of certain parameters are discussed with several numerical examples.

辽东栎林演替的生理生态研究

通过野外测定和控制实验结果发现： 在成林内林窗能促进发芽率、幼苗存活率和光合作用等。高光下,会发生多次生长。幼苗干物质积累与光照明显正相关。生长速度与存活无相关，但对第二年存活有影响。基径可作为光反应的无损伤测定指标。直射光和散射光对幼苗生长影响不同。 适当厚度的枯落物覆盖促进种子发芽和补充更新,埋藏太深则对辽东栎幼苗的生长、生理和存活等有抑制。灌木和树木的冠层对辽东栎在灌丛中的建立。 模拟实验表明完全去叶可导致16.7%的幼苗死亡，完全去除子叶可导致50%幼苗死亡，都严重影响幼苗的生理过程和干物质积累。土壤养分和干旱对辽东栎幼苗生长和光合也有很大影响。 运用通径分析精确测定,结果发现辽东栎冠层不同位置的叶片气体交换的限制因子有很大的差别。上层叶主要是气孔导度影响,叶温是通过饱和蒸汽压（VPD）有间接负作用。冠层下方光照的不足的限制,叶温有促进光合作用。不同部位叶片光合作用对气孔导度的依赖程度不同。 在辽东栎林演替过程中，生理生态变化趋势明显。根据所测的20多个生理指标,可将主要物种划分成不同的功能群。早期种具有较高的光合速率,气孔导度, 蒸腾速率和抗旱性, 而辽东栎和棘皮桦单独为一类。

L'Organisation de la famille, selon le vrai modèle signalé par l'histoire de toutes les races et de tous les temps, par M. F. Le Play,... avec trois appendices, par MM. É. Cheysson, F. Le Play et C. Jannet. 2e édition...

Comprend : La Famille-souche du Lavedan

Libro-Objeto.

Se trata de una experiencia desarrollada por el CEP de Móstoles y aplicada en varios centros públicos de dicho municipio. El proyecto de Libro-Objeto es una propuesta para investigar y experimentar a través de las áreas de Expresión Artística y Lengua. Utiliza el libro como recurso didáctico considerado desde el punto de vista material (continente) e intelectual (contenido). Los objetivos, que atienden al área de expresión artística y área de lenguaje, son: utilizar los recursos básicos de los diferentes lenguajes (plástico, dramático y musical) que permitan al alumno expresarse y comunicarse creativamente con los demás; desarrollar armónicamente las capacidades y habilidades de expresión plástica, dramática y musical; utilizar la lengua escrita con soltura y corrección; y crear hábitos de lectura. La experiencia se desarrolla a través del diseño y confección de libros en relieve y con diferentes formas y materiales. Para el contenido de esos libros se crean historias, personajes, canciones y poesías. Todo este trabajo creativo se presenta al resto de la comunidad educativa con una exposición y con la celebración de un festival de obras dramáticas y musicales. La valoración del proyecto es positiva ya que permite la participación activa de los alumnos.

Development of successive cambia and structure of wood in Gallesia integrifolia (Spreng.) Harms (Phytolaccaceae)

Stem diameter in Gallesia integrifolia (Spreng.) Harms (Phytolaccaceae) increases by forming concentric rings of xylem alternating with phloem, which show frequent anastomoses. After a period of primary growth and the formation of first (normal) ring of vascular cambium, further successive rings are initiated outside this cambium. The second ring of cambium originates from the pericycle parenchyma located between the proto-phloem, and the pericycle fibres. Each cambium produces centripetally secondary xylem and centrifugally secondary phloem. Differentiation of xylem precedes that of phloem and the first elements formed are always xylem fibres. Structurally, the vascular cylinder is composed by successive rings of secondary xylem and phloem. These rings are separated by wide bands of conjunctive parenchyma tissue. Presence of collateral vascular bundles with irregular orientation is observed in the region of anastomoses of two or more bands of conjunctive tissue. These bundles are surrounded by isodiametric, lignified and thick-walled cells. In some of the cambial rings, occurrence of polycentric rays was also noticed; these rays are tall, and characterized by the presence of meristematic regions that differentiated into thick-walled elements of secondary xylem. Origin and development of the successive cambia and the structure of xylem are discussed.

Blood parameters of chicks from eggs injected with ascorbic acid and subjected to thermal stress

This study was aimed to verify if chicks from eggs injected with ascorbic acid and subjected to thermal stress would have higher immunity than chicks from incubation at thermoneutrality without injection of ascorbic acid. The parameters evaluated were temperature on oxygen saturation in hemoglobin, glucose, number of erythrocytes, hematocrit rate and number of hemoglobins of newly hatched male chicks, hatched from eggs injected with ascorbic acid (AA) and subjected to thermal stress during incubation. The experimental design was completely randomized in factorial scheme 5 (application levels of ascorbic acid) x 2 (incubation temperatures). The data were subjected to analysis of variance using the General Linear Model procedure (GLM) of SAS ®. For the parameters (number of erythrocytes, rate of hematrocit and values of hemoglobin), there was significant interaction (p <0.05) between treatments in egg and incubation temperatures. Analyzing the interactions for these parameters, it was observed that the application of 0% ascorbic acid in egg minimized the effect of heat stress when compared with treatment without injection. The application of ascorbic acid levels in eggs incubated under heat stress failed to maximize the immunity of newly hatched chicks. It is assumed that the increased liquid in the amniotic fluid, in those embryos injected with water, favored the lower heat conductance for these embryos, thus helping in their development in relation to immunity. Considering that hemoglobin is related to the transport of gases, these data suggest that increasing the concentration of AA solution inoculated may influence the respiratory rates of eggs.

Cryopreservation of umbilical cord mesenchymal cells in xenofree conditions

Background aims. Mesenchymal stromal cells (MSC) are being used to treat and prevent a variety of clinical conditions. To be readily available, MSC must be cryopreserved until infusion. However, the optimal cryopreservation methods, cryoprotector solutions and MSC sensitivity to dimethyl sulfoxide (DMSO) exposure are unknown. This study investigated these issues. Methods. MSC samples were obtained from human umbilical cord (n = 15), expanded with Minimal Essential Medium-alpha (alpha-MEM) 10% human serum (HS), resuspended in 25 mL solution (HS, 10% DMSO, 20% hydroxyethyl starch) and cryopreserved using the BioArchive (R) system. After a mean of 18 +/- 7 days, cell suspensions were thawed and diluted until a DMSO concentration of 2.5% was reached. Samples were tested for cell quantification and viability, immunophenotype and functional assays. Results. Post-thaw cell recovery: 114 +/- 2.90% (mean +/- SEM). Recovery of viable cells: 93.46 +/- 4.41%, 90.17 +/- 4.55% and 81.03 +/- 4.30% at 30 min, 120 min and 24 h post-thaw, respectively. Cell viability: 89.26 +/- 1.56%, 72.71 +/- 2.12%, 70.20 +/- 2.39% and 63.02 +/- 2.33% (P<0.0001) pre-cryopreservation and 30 min, 120 min and 24 h post-thaw, respectively. All post-thaw samples had cells that adhered to culture bottles. Post-thaw cell expansion was 4.18 +/- 0.17 X, with a doubling time of 38 +/- 1.69 h, and their capacity to inhibit peripheral blood mononuclear cells (PBMC) proliferation was similar to that observed before cryopreservation. Differentiation capacity, cell-surface marker profile and cytogenetics were not changed by the cryopreservation procedure. Conclusions. A method for cryopreservation of MSC in bags, in xenofree conditions, is described that facilitates their clinical use. The MSC functional and cytogenetic status and morphologic characteristics were not changed by cryopreservation. It was also demonstrated that MSC are relatively resistant to exposure to DMSO, but we recommend cell infusion as soon as possible.