Transduction of an immortalized olfactory ensheathing glia cell line with the green fluorescent protein (GFP) gene: Evaluation of its neuroregenerative capacity as a proof of concept.

Olfactory ensheathing glia (OEG) cells are known to foster axonal regeneration of central nervous system (CNS) neurons. Several lines of reversibly immortalized human OEG (ihOEG) have been previously established that enabled to develop models for their validation in vitro and in vivo. In this work, a constitutively GFP-expressing ihOEG cell line was obtained, and named Ts14-GFP. Ts14-GFP neuroregenerative ability was similar to that found for the parental line Ts14 and it can be assayed using in vivo transplantation experimental paradigms, after spinal cord or optic nerve damage. Additionally, we have engineered a low-regenerative ihOEG line, hTL2, using lentiviral transduction of the large T antigen from SV40 virus, denominated from now on Ts12. Ts12 can be used as a low regeneration control in these experiments.pre-print281 K

Reflexión de futuros profesores de matemáticas sobre las dificultades del aprendizaje algebraico

En el contexto de la formación inicial de futuros profesores de matemáticas (FPM), se analiza la reflexión de dos futuros profesores sobre los errores y las dificultades del aprendizaje algebraico. Con este objetivo se diseñó e implementó un módulo formativo en el prácticum de un programa de licenciatura en matemáticas en Colombia. La investigación se configuró bajo la teoría del aprendizaje realista para promover la reflexión, en coherencia con este marco teórico se orientó la realización de un ciclo reflexivo bajo el modelo ALaCT. Siguiendo el enfoque de la investigación cualitativa, de carácter interpretativo y usando el análisis de contenido se han examinado las producciones de los participantes, los registros obtenidos en la intervención y del diario de campo del investigador. Los resultados muestran, que los futuros profesores llevaron a cabo cinco fases de reflexión, en las cuales el análisis de los errores y las dificultades del aprendizaje algebraico transita por mínimo cuatro momentos, que implican a los futuros profesores en decisiones y alternativas para abordar la instrucción. Se informa cómo evolucionó este análisis y el conocimiento profesional involucrado para fundamentarlo

Increased migration of olfactory ensheathing cells secreting the Nogo receptor ectodomain over inhibitory substrates and lesioned spinal cord

Abstract Olfactory ensheathing cell (OEC) transplantation emerged some years ago as a promising therapeutic strategy to repair injured spinal cord. However, inhibitory molecules are present for long periods of time in lesioned spinal cord, inhibiting both OEC migration and axonal regrowth. Two families of these molecules, chondroitin sulphate proteoglycans (CSPG) and myelin-derived inhibitors (MAIs), are able to trigger inhibitory responses in lesioned axons. Mounting evidence suggests that OEC migration is inhibited by myelin. Here we demonstrate that OEC migration is largely inhibited by CSPGs and that inhibition can be overcome by the bacterial enzyme Chondroitinase ABC. In parallel, we have generated a stable OEC cell line overexpressing the Nogo receptor (NgR) ectodomain to reduce MAI-associated inhibition in vitro and in vivo. Results indicate that engineered cells migrate longer distances than unmodified OECs over myelin or oligodendrocyte-myelin glycoprotein (OMgp)-coated substrates. In addition, they also show improved migration in lesioned spinal cord. Our results provide new insights toward the improvement of the mechanisms of action and optimization of OEC-based cell therapy for spinal cord lesion.Spanish Ministry of Science and Innovation (BFU2012-32617), the Generalitat de Catalunya (SGR2014-1218), La Caixa Obra Social Foundation, and the Basque Foundation of Health and Innovation Research (BIO12/AL/004) to JADR. RG was supported by Fondo de Investigaciones Sanitarias (PI11-00075) and work in FW’s lab was supported by grants from the Dirección General de Ciencia y Tecnologia-DGCYT-(SAF2012-39148-C03-01), and EU-FP7-2009-(CT222887), as well as an institutional grant from the ‘Fundación ArecesPeer Reviewe

Coculture of Axotomized Rat Retinal Ganglion Neurons with Olfactory Ensheathing Glia, as an In Vitro Model of Adult Axonal Regeneration.

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Reflexión en el prácticum: Un experimento de enseñanza con estudiantes colombianos

Resumen:La formación inicial del profesorado de matemáticas afronta el reto de orientar a los futuros profesores para que vean con sentido el conocimiento teórico, al enfrentar problemas prácticos del contexto profesional. Una forma de favorecer esta articulación teoría-práctica es mediante el ejercicio de hábitos de reflexión, especialmente en el prácticum. En este contexto llevamos a cabo un experimento de enseñanza en el curso de práctica docente (o prácticum) con 12 estudiantes de último año de Licenciatura en Matemáticas de la Universidad de los Llanos (Colombia). Dicho experimento es el estudio empírico de una investigación doctoral, uno de sus objetivos es promover la reflexión de futuros profesores de matemáticas sobre aspectos de su práctica profesional. Para ello, se describe de manera sistemática el experimento de enseñanza, atendiendo a las etapas de preparación y de experimentación de la instrucción que siguen el paradigma de la investigación de diseño. La revisión y el análisis secuencial de cada módulo durante la experimentación han permitido identificar algunas cualidades de la instrucción, en la que se promueve la realización de ciclos reflexivos. De igual modo, el experimento facilitó a los futuros profesores conocer fundamentos, aplicar procedimientos y aprovechar recursos que, fundamentados en la didáctica de la matemática, les permitieron diseñar, implementar y evaluar actividades de enseñanza y aprendizaje para su clase. Abstract:The preservice mathematics teacher’s training faces the complexity for future teachers that they see the theoretical knowledge related to practical problems facing their professional context. One way to promote this theoretical and practical articulation is by exercising habits of reflection, especially in the practicum. In this context, we conducted a teaching experiment in the course of teaching practice (or practicum) with 12 students last year of Degree in Mathematics of the Llanos University (Colombia). This experiment is the empirical study of a research, which has one objective to promote reflection in future math teachers. Systematically describe the teaching experiment, taking the stages of preparation and experimentation of instruction according to the paradigm of design research. Review and sequential analysis to module during experimentation has identified some qualities of instruction regarding the conduct of reflective cycles. Similarly, the experiment provided to future teachers know fundaments, implement procedures and leverage resources, based on the teaching of mathematics, allowed

Spanish Nursing Students' Practical Experience of Computer Applications in Nursing

This study aimed to determine the level of computer practical experience in a sample of Spanish nursing students. Each student was given a Spanish language questionnaire, modified from an original used previously with medical students at the Medical School of North Carolina University (USA) and also at the Education Unit of Hospital General Universitario del Mar (Spain). The 10-item self-report questionnaire probed for information about practical experience with computers. A total of 126 students made up the sample. The majority were female (80.2%; n=101). The results showed that just over half (57.1%, n=72) of the students had used a computer game (three or more times before), and that only one third (37.3%, n=47) had the experience of using a word processing package. Moreover, other applications and IT-based facilities (e.g. statistical packages, e-mail, databases, CD-ROM searches, programming languages and computer-assisted learning) had never been used by the majority of students. The student nurses' practical experience was less than that reported for medical students in previous studies

Olfactory ensheathing cell-conditioned medium reverts 3 Ab25–35-induced oxidative damage in SH-SY5Y cells 4 by modulating the mitochondria-mediated apoptotic pathway.

Olfactory ensheathing cells (OECs) are a type of 10 glia from the mammalian olfactory system, with neuro- 11 protective and regenerative properties. b-Amyloid peptides 12 are a major component of the senile plaques characteristic 13 of the Alzheimer brain. The amyloid beta (Ab) precursor 14 protein is cleaved to amyloid peptides, and Ab25–35 is 15 regarded to be the functional domain of Ab, responsible for 16 its neurotoxic properties. It has been reported that Ab25–35 17 triggers reactive oxygen species (ROS)-mediated oxidative 18 damage, altering the structure and function of mitochon- 19 dria, leading to the activation of the mitochondrial intrinsic 20 apoptotic pathway. Our goal is to investigate the effects of 21 OECs on the toxicity of aggregated Ab25–35, in human 22 neuroblastoma SH-SY5Y cells. For such purpose, SH- 23 SY5Y cells were incubated with Ab25–35 and OEC-conditioned medium (OECCM). OECCM promoted the 24 cell viability and reduced the apoptosis, and decreased the 25 intracellular ROS and the lipid peroxidation. In the pres- 26 ence of OECCM, mRNA and protein levels of antioxidant 27 enzymes (SOD1 and SOD2) were upregulated. Concomi- 28 tantly, OECCM decreased mRNA and the protein expres- 29 sion levels of cytochrome c, caspase-9, caspase-3, and Bax 30 in SH-SY5Y cells, and increased mRNA and the protein 31 expression level of Bcl-2. However, OECCM did not alter 32 intracellular Ca 33 2? concentration in SH-SY5Y cells. Taken together, our data suggest that OECCM ameliorates 34 Ab25–35-induced oxidative damage in neuroblastoma SH- 35 SY5Y cells by inhibiting the mitochondrial intrinsic path- 36 way. These data provide new insights into the functional 37 actions of OECCM on oxidative stress-induced cell 38 damage.pre-print2138 K

Cell therapy for spinal cord injury with Olfactory Ensheathing Glia Cells (OECs)

This is the peer reviewed version of the following article: Gómez RM, Sánchez MY, Portela-Lomba M, et al. Cell therapy for spinal cord injury with olfactoryensheathing glia cells (OECs). Glia. 2018;00:1–35 which has been published in final form at GLIA 13 January (2018) http://doi.org/10.1002/glia.23282. © 2018 Wiley Periodicals This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.The prospects of achieving regeneration in the central nervous system (CNS) have changed, asmost recent findings indicate that several species, including humans, can produce neurons in adult-hood. Studies targeting this property may be considered as potential therapeutic strategies torespond to injury or the effects of demyelinating diseases in the CNS. While CNS trauma mayinterrupt the axonal tracts that connect neurons with their targets, some neurons remain alive, asseen in optic nerve and spinal cord (SC) injuries (SCIs). The devastating consequences of SCIs aredue to the immediate and significant disruption of the ascending and descending spinal pathways,which result in varying degrees of motor and sensory impairment. Recent therapeutic studies forSCI have focused on cell transplantation in animal models, using cells capable of inducing axonregeneration like Schwann cells (SchCs), astrocytes, genetically modified fibroblasts and olfactoryensheathing glia cells (OECs). Nevertheless, and despite the improvements in such cell-based ther-apeutic strategies, there is still little information regarding the mechanisms underlying the successof transplantation and regarding any secondary effects. Therefore, further studies are needed to clarify these issues. In this review, we highlight the properties of OECs that make them suitable toachieve neuroplasticity/neuroregeneration in SCI. OECs can interact with the glial scar, stimulateangiogenesis, axon outgrowth and remyelination, improving functional outcomes following lesion.Furthermore, we present evidence of the utility of cell therapy with OECs to treat SCI, both fromanimal models and clinical studies performed on SCI patients, providing promising results for future treatments

Human placenta-derived mesenchymal stem cells stimulate neuronal regeneration by promoting axon growth and restoring neuronal activity

In the last decades, mesenchymal stem cells (MSCs) have become the cornerstone of cellular therapy due to their unique characteristics. Specifically human placenta-derived mesenchymal stem cells (hPMSCs) are highlighted for their unique features, including ease to isolate, non-invasive techniques for large scale cell production, significant immunomodulatory capacity, and a high ability to migrate to injuries. Researchers are exploring innovative techniques to overcome the low regenerative capacity of Central Nervous System (CNS) neurons, with one promising avenue being the development of tailored mesenchymal stem cell therapies capable of promoting neural repair and recovery. In this context, we have evaluated hPMSCs as candidates for CNS lesion regeneration using a skillful co-culture model system. Indeed, we have demonstrated the hPMSCs ability to stimulate damaged rat-retina neurons regeneration by promoting axon growth and restoring neuronal activity both under normoxia and hypoxia conditions. With our model we have obtained neuronal regeneration values of 10%–14% and axonal length per neuron rates of 19-26, μm/neuron. To assess whether the regenerative capabilities of hPMSCs are contact-dependent effects or it is mediated through paracrine mechanisms, we carried out transwell co-culture and conditioned medium experiments confirming the role of secreted factors in axonal regeneration. It was found that hPMSCs produce brain derived, neurotrophic factor (BDNF), nerve-growth factor (NGF) and Neurotrophin-3 (NT-3), involved in the process of neuronal regeneration and restoration of the physiological activity of neurons. In effect, we confirmed the success of our treatment using the patch clamp technique to study ionic currents in individual isolated living cells demonstrating that in our model the regenerated neurons are electrophysiologically active, firing action potentials. The outcomes of our neuronal regeneration studies, combined with the axon-regenerating capabilities exhibited by mesenchymal stem cells derived from the placenta, present a hopeful outlook for the potential therapeutic application of hPMSCs in the treatment of neurological disorders.post-print2885 K

Small molecules fail to induce direct reprogramming of adult rat olfactory ensheathing glia to mature neurons

An approach to generate new neurons after central nervous system injury or disease is direct reprogramming of the individual's own somatic cells into differentiated neurons. This can be achieved either by transduction of viral vectors that express neurogenic transcription factors and/or through induction with small molecules, avoiding introducing foreign genetic material in target cells. In this work, we propose olfactory ensheathing glia (OEG) as a candidate for direct reprogramming to neurons with small molecules due to its well-characterized neuro-regenerative capacity. After screening different combinations of small molecules in different culture conditions, only partial reprogramming was achieved: induced cells expressed neuronal markers but lacked the ability of firing action potentials. Our work demonstrates that direct conversion of adult olfactory ensheathing glia to mature, functional neurons cannot be induced only with pharmacological tools