Diabetes mellitus: Lessons from COVID-19 for monkeypox infection.

BACKGROUND AND AIMS: Type 2 Diabetes Mellitus is known to be linked to malfunctioning antiviral defense; however, its association with the severity of monkeypox is poorly understood. In this review, we discuss key immunological mechanisms in the antiviral response affected by poor glucose control that could impact the susceptibility and severity of monkeypox infection, leading to a heightened emphasis on the use of the available antidiabetic drugs. METHODS: We searched PubMed and Google scholar for articles published from January 1985 to August 2022. No criteria for publication data were set, and all articles in English were included. RESULTS: Currently, there are no studies about the risk or consequences of monkeypox infection in the diabetic population. A high incidence of diabetes is reported in countries such as China, India, Pakistan, EUA, Indonesia, Brazil, Mexico, Bangladesh, Japan, and Egypt, where unfortunately imported cases of monkeypox have been reported and the infection continues to spread. CONCLUSIONS: High incidence of diabetes together with the cessation of smallpox vaccination has left large numbers of the human population unprotected against monkeypox. The best option for the population remains confined to the prevention of infection as well as the use of hypoglycemic agents that have also been shown to improve immune mechanisms associated with viral protection.

Encoding signs of orofacial neuropathic pain from facial expressions in mice.

OBJECTIVE: To evaluate the effects of mental nerve injury in the facial reactions elicited by mechanical stimulation of different intensities and detect and quantify spontaneous facial pain-like expressions during a period free of stimuli, as signs of evoked and spontaneous pain in a mouse model for neuropathic orofacial pain. DESIGN: We recorded mouse heads in a fixed position during a stimulus-free period and with mechanical stimulation with 3 different Von Frey filaments. We extracted the Histograms of Oriented Gradients of each frame of the video recordings to be compared with a prototypical pain-like facial expression. The similarity score was then used to register and quantify the percentage of spontaneous pain-like facial reactions and evaluate the increased similarity to the prototypical pain-like face evoked by mechanical stimuli. The assessments were made one day before and four days after a unilateral mental nerve compression. RESULTS: Our findings show that mental nerve injury promotes an increase in spontaneous facial pain-like expressions and reduced mechanical threshold, reflected in a higher similarity to our pain-like face prototype, regardless of the intensity of the stimuli applied. CONCLUSIONS: Machine vision encodes the facial expression associated with evoked and spontaneous pain after mental nerve injury for up to four days. Facial expression quantitatively reflects the increased mechanical sensitivity elicited by mental nerve injury. We also show that this technique can detect spontaneous pain-like responses from facial reactions. Artificial vision can be applied to evaluate signs of orofacial neuropathic pain to study the involved neural circuits.

Functionalised peptide hydrogel for the delivery of cardiac progenitor cells.

Heart failure (HF) remains one of the leading causes of death worldwide; most commonly developing after myocardial infarction (MI). Since adult cardiomyocytes characteristically do not proliferate, cells lost during MI are not replaced. As a result, the heart has a limited regenerative capacity. There is, therefore, a need to develop novel cell-based therapies to promote the regeneration of the heart after MI. The delivery and retention of cells at the injury site remains a significant challenge. In this context, we explored the potential of using an injectable, RGDSP-functionalised self-assembling peptide - FEFEFKFK - hydrogel as scaffold for the delivery and retention of rat cardiac progenitor cells (CPCs) into the heart. Our results show that culturing CPCs in vitro within the hydrogel for one-week promoted their spontaneous differentiation towards adult cardiac phenotypes. Injection of the hydrogel on its own, or loaded with CPCs, into the rat after injury resulted in a significant reduction in myocardial damage and left ventricular dilation.

Análisis del conocimiento y manejo de la fiebre por parte de pediatras y residentes en relación a un proceso asistencial establecido / Analysis of pediatricians' and residents' knowledge of fever management in relation to established guidelines

Objetivo: evaluar el consejo dado a padres sobre la fiebre y conocer la incidencia estimada de fiebre sin foco (FSF) en consulta, la accesibilidad a exámenes complementarios (EC) y la aplicación de un protocolo (PF). Métodos: cuestionario sobre un total de 151 pediatras de Atención Primaria (PAP), pediatras de hospital (PH) y residentes de Pediatría (R). Se utiliza el paquete estadístico SPSS® 15.0. Para variables cualitativas el test de la χ2, siendo el valor significativo p < 0,05. Resultados: se han evaluado 109 cuestionarios: mujeres 65,4% y hombres 34,6%. El 44,9% definió como fiebre 37,5 °C en axila y 38 °C en recto. El 78,9% aconseja termómetro electrónico; el 93,6%, medidas físicas; el 79,8%, paracetamol, y el 76,1% alterna antitérmicos en casos seleccionados. El 56,2% diagnostica un 10% de FSF a la semana, el 19,3% codifica siempre; el 31,2% algunas veces, y el 45,9% no lo hace. En menores de seis meses, el 91,7% solicita tira de orina, y el 41,3%, urocultivo; En pacientes de 6-12 meses, el 96,3% solicita tira de orina, y el 11,9%, urocultivo. Los PAP reciben resultados el mismo día: hemograma (3%) y radiografía (68,6%), en menos de 72 horas: urocultivo (38,7%). Los PH y R reciben el mismo día: hemograma (88,3%) y radografía (85,7%); en menos de 72 horas: urocultivo (85,7%). El 74,6% de los PAP deriva al hospital a los menores de tres meses con FSF, el 64,7% de los PH y el 83,3% de los R hacen EC. Conoce el PF el 78,9%, de los cuales, el 69,8% cree que es aplicable y, a su vez, lo aplica un 65,4%. Conclusiones: consejo mayoritario de termómetro electrónico, uso de medidas físicas y paracetamol. Alternancia seleccionada de antitérmicos. Bajo diagnóstico y codificación de FSF. Limitado acceso a exámenes complementarios para PAP. Alto conocimiento del PF pero baja aplicación (AU)

Objective: to evaluate the advice given to parents about fever, determine the incidence of Fever Without Source (FWS) in the pediatrician’s office, availability of complementary examinations (CE) and the implementation of fever guidelines (FG). Methods: questionnaires distributed to 151 Primary Attention Pediatricians (PAP), Hospital Pediatricians (HP) and Residents (R). Statistical analysis SPSS 15.0. For qualitative variables the 2 test was used. Significant value was p < 0.05. Results: 109 questionnaires analyzed: women 65.4% and men 34.6%. 44.9% defined fever 37.5 °C axilar temperature and 38 °C rectal temperature. 78.9% advise the use of electronic thermometer, 93.6% advice taking non-drug measures, 79.8% choose paracetamol and 76.1% alternate antipyretics in selected cases. 56.2% diagnosed 10% of FWS per week, 19.3% always encode it, 31.2% sometimes and 45.9% never. For infants < 6 m are required: 91.7% urine strips, 41.3% urine culture; from 6-12 months: 96.3% urine strips and 11.9% urine culture. PAP receive results the same day: CBC count 3%, radiology 68.6% and urine culture in less than 72h, 38.7%. HP and R the same day, CBC (AU)

Conocimiento y actuación de los padres sobre la fiebre / Knowledge and management of fever by parents

Objetivos: evaluar los conocimientos y actitudes que tienen los padres sobre la fiebre, así como la influencia de los aspectos familiares. Material y métodos: cuestionario distribuido a padres de dos áreas asistenciales con hijos de 1-5 años. Para variables cualitativas se aplicaron pruebas de asociación mediante el test X2; para las variables cuantitativas se aplicó la diferencia de medias mediante la t de Student o análisis de la varianza (ANOVA). Se consideró como valor estadísticamente significativo p < 0,05. Resultados: se analizaron 288 cuestionarios. El 50% de los encuestados tiene dos hijos. Trabaja el 64,5%. En el área urbana son de mayor edad y nivel de estudios (p < 0,001). Un 50,3% considera la fiebre mala, menos los de edad media superior (p < 0,05). El 67,7% utiliza termómetro electrónico. Consideran fiebre una temperatura de 37,7 ºC en axila. Ante la fiebre, el 58,3% utiliza en primer lugar un antitérmico. El 98,2% usa medidas físicas y el 49,3% de ellos piensa que mejoran la fiebre; las usan menos los que trabajan (p < 0,05). Los de estudios superiores quitan ropa y dan líquidos más que los de estudios primarios (p = 0,035). Los antitérmicos más empleados fueron paracetamol e ibuprofeno. Un 64,6% de los encuestados percibe diferencias en cuanto a eficacia. El 85,4% utiliza la dosis indicada por su pediatra y el 21,5%, la que indica la ficha técnica, sobre todo los de estudios superiores frente a los de estudios primarios (p < 0,05). El 67,4% alterna antitérmicos, siempre aconsejados por el pediatra. Conclusiones: globalmente, en la población estudiada existe un buen conocimiento y una actitud adecuada ante la fiebre (AU)

Objective: To assess parental knowledge and attitudes about fever and the influence of social and family aspects. Methods: Questionnaires distributed to parents of children 1-5 years old in two health districts. For qualitative variables association tests with X2 test were applied, and mean differences by Student’s t-distribution or analysis of variance (ANOVA) were used for quantitative variables. It was considered statistically significant the value of p < 0.05. Results: There were 288 questionnaires analyzed. Fifty percent of respondents have 2 children, and 64.5% work. Older age and higher education levels were found in urban areas (p < 0.001). Fever was considered to be a bad thing by 50.3%, less so those with higher mean age (p < 0.05). Electronic thermometers was used in 67.7%, and 86.2% took armpit temperature considering 37.7 ºC as fever. When faced with fever, 58.3% of parents first use antipyretics. Physical measures are used in the first term by 98.2% and 49.3% think these measures lower the fever; they are used less by those parents who work (p < 0.05). Parents with higher education levels remove the clothing and give liquids more than those with primary education (p = 0.035). Most commonly used antipyretics are acetaminophen and ibuprofen; 64.6% perceived differences in efficiency; 65.4% think that ibuprofen is more effective than acetaminophen. Most parents use the dosage prescribed by the pediatrician (85.4%), and 21.5% use the dosage specified in the leaflet, especially those with higher education levels, compared to parents with primary education (p < 0.05). They sometimes alternate antipyretics (67.4%), always following the advice of their pediatrician. Conclusions: There is an overall good knowledge and attitudes about fever (AU)

Neurogénesis como diana terapéutica para la enfermedad de Alzheimer / Neurogenesis as a therapeutic target for alzheimer´s disease

Introducción. El cerebro adulto de mamíferos conserva la capacidad de generar nuevas neuronas a partir de células troncales/progenitoras neuronales. Las nuevas neuronas se integran a las redes preexistentes a través de un proceso denominado ‘neurogénesis en el cerebro adulto’, que está confinado a regiones del cerebro con un alto grado de plasticidad y asociadas a funciones que muestran deterioro en la enfermedad de Alzheimer. Desarrollo. A pesar de lo conocido, permanecen muchos interrogantes alrededor de estos fenómenos neurogénicos, su regulación y su potencial terapéutico real en neurodegeneraciones como la referida. Conclusiones. Hemos revisado el tema de la neurogénesis del cerebro adulto desde el punto de vista preclínico (modelado experimental) y terapéutico en el marco de la enfermedad de Alzheimer (AU)

Introduction. The adult brain of mammals preserves the capacity to generate new neurons from neural stem/ progenitor cells. The new neurons become part of the already-existing networks by means of a process called ‘neurogenesis in the adult brain’, which is restricted to regions of the brain with a high degree of plasticity and which are associated to functions that are impaired in Alzheimer’s disease. Development. Despite increasing knowledge, there are still many questions surrounding these neurogenic phenomena, their regulation and their real therapeutic potential in cases of neurodegeneration such as the one referred to here. Conclusions. We have reviewed the subject of neurogenesis of the adult brain from both the pre-clinical point of view (experimental modelling) and the therapeutic perspective within the framework of Alzheimer’s disease (AU)

[Neurogenesis as a therapeutic target for Alzheimer's disease]. / Neurogénesis como diana terapéutica para la enfermedad de Alzheimer.

INTRODUCTION: The adult brain of mammals preserves the capacity to generate new neurons from neural stem/progenitor cells. The new neurons become part of the already-existing networks by means of a process called 'neurogenesis in the adult brain', which is restricted to regions of the brain with a high degree of plasticity and which are associated to functions that are impaired in Alzheimer's disease. DEVELOPMENT: Despite increasing knowledge, there are still many questions surrounding these neurogenic phenomena, their regulation and their real therapeutic potential in cases of neurodegeneration such as the one referred to here. CONCLUSIONS: We have reviewed the subject of neurogenesis of the adult brain from both the pre-clinical point of view (experimental modelling) and the therapeutic perspective within the framework of Alzheimer's disease.

[Evaluation of the survival of bone marrow mononucleate cells transplanted in a rat model of striatal lesion with quinolinic acid]. / Evaluacion de la supervivencia de las células mononucleadas de la médula ósea trasplantadas en un modelo de ratas con lesión estriatal por ácido quinolínico.

INTRODUCTION: Transplant is one of the alternatives available for the treatment of neurodegenerative diseases aimed at replacing the cells lost during the course of the disease. One promising source of cells for the development of transplants could be the mononucleate cells from bone marrow. AIMS. The purpose of this study was to study the capacity of bone marrow mononucleate cells to survive the transplant process, and to search for a method that enables tracking of these cells in vivo once they have been implanted. MATERIALS AND METHODS: Bone marrow mononucleate cells were extracted from the femur of rats by means of a Ficoll-Hypaque gradient. The cells under study were modified genetically with an adenovirus that expresses the PFV or which are marked with Hoechst dye. The marked cells were implanted in the striatum of rats with lesions caused by quinolinic acid. RESULTS: The viability of the genetically modified cells was low, whereas that of the cells marked with Hoechst dye was above 90%. The implanted cells survived the transplant at least a month and dispersed away from the site of entry towards the corpus callosum and cortex. CONCLUSIONS: We consider that the use of Hoechst dye offers more advantages for tracking these cells in vivo. Mononucleate cells have a number of characteristics that allow them to be included as candidate sources of cells for the treatment of neurodegenerative diseases.

Evaluación de la supervivencia de las células mononucleadas de la médula ósea trasplantadas en un modelo de ratas de lesión estriatal con ácido quinolínico / Evaluation of the survival of bone marrow mononucleate cells transplanted in a rat model of striatal lesion with quinolinic acid

Introducción. El trasplante es una de las alternativas para el tratamiento de enfermedades neurodegenerativas, y está encaminado hacia el reemplazo de las células perdidas durante el desarrollo de la enfermedad. Una fuente celular prometedora para el desarrollo de los trasplantes podrían ser las células mononucleadas de la médula ósea. Objetivo. Estudiar la capacidad de las células mononucleadas de la médula ósea de sobrevivir al trasplante y buscar un método que permita el seguimiento de estas células in vivo una vez implantadas. Materiales y métodos. Las células mononucleadas fueron extraídas del fémur de ratas mediante un gradiente de Ficoll-Hypaque. Las células objeto de estudio fueron modificadas genéticamente con un adenovirus que expresa la PFV o marcadas con el reactivo de Hoechst. Las células marcadas se implantaron en el estriado de ratas lesionadas con ácido quinolínico. Resultados. La viabilidad de las células modificadas genéticamente fue baja, mientras que la de las células marcadas con el reactivo de Hoechst fue superior al 90 por ciento. Las células implantadas sobrevivieron al trasplante al menos un mes y se dispersaron desde el sitio de entrada hacia el cuerpo calloso y la corteza. Conclusiones. Consideramos más ventajoso el uso del reactivo de Hoechst para el seguimiento de estas células in vivo. Las células mononucleadas tienen características que les permiten formar parte de las fuentes celulares candidatas para el tratamiento de las enfermedades neurodegenerativas(AU)

Introduction: Transplant is one of the alternatives available for the treatment of neurodegenerative diseases aimed at replacing the cells lost during the course of the disease. One promising source of cells for the development of transplants could be the mononucleate cells from bone marrow. AIMS. The purpose of this study was to study the capacity of bone marrow mononucleate cells to survive the transplant process, and to search for a method that enables tracking of these cells in vivo once they have been implanted. MATERIALS AND METHODS: Bone marrow mononucleate cells were extracted from the femur of rats by means of a Ficoll-Hypaque gradient. The cells under study were modified genetically with an adenovirus that expresses the PFV or which are marked with Hoechst dye. The marked cells were implanted in the striatum of rats with lesions caused by quinolinic acid. RESULTS: The viability of the genetically modified cells was low, whereas that of the cells marked with Hoechst dye was above 90percent. The implanted cells survived the transplant at least a month and dispersed away from the site of entry towards the corpus callosum and cortex. CONCLUSIONS: We consider that the use of Hoechst dye offers more advantages for tracking these cells in vivo. Mononucleate cells have a number of characteristics that allow them to be included as candidate sources of cells for the treatment of neurodegenerative diseases

Evaluación de la supervivencia de las células mononucleadas de la médula ósea trasplantadas en un modelo de ratas con lesión estriatal por ácido quinolínico / Evaluation of the survival of bone marrow mononucleate cells transplanted in a rat model of striatal lesion with quinolinic acid

Introducción. El trasplante es una de las alternativas para el tratamiento de enfermedades neurodegenerativas, y está encaminado hacia el reemplazo de las células perdidas durante el desarrollo de la enfermedad. Una fuente celular prometedora para el desarrollo de los trasplantes podrían ser las células mononucleadas de la médula ósea. Objetivo. Estudiar la capacidad de las células mononucleadas de la médula ósea de sobrevivir al trasplante y buscar un método que permita el seguimiento de estas células in vivo una vez implantadas. Materiales y métodos. Las células mononucleadas fueron extraídas del fémur de ratas mediante un gradiente de Ficoll-Hypaque. Las células objeto de estudio fueron modificadas genéticamente con un adenovirus que expresa la PFV o marcadas con el reactivo de Hoechst. Las células marcadas se implantaron en el estriado de ratas lesionadas con ácido quinolínico. Resultados. La viabilidad de las células modificadas genéticamente fue baja, mientras que la de las células marcadas con el reactivo de Hoechst fue superior al 90%. Las células implantadas sobrevivieron al trasplante al menos un mes y se dispersaron desde el sitio de entrada hacia el cuerpo calloso y la corteza. Conclusiones. Consideramos más ventajoso el uso del reactivo de Hoechst para el seguimiento de estas células in vivo. Las células mononucleadas tienen características que les permiten formar parte de las fuentes celulares candidatas para el tratamiento de las enfermedades neurodegenerativas

Introduction. Transplant is one of the alternatives available for the treatment of neurodegenerative diseases aimed at replacing the cells lost during the course of the disease. One promising source of cells for the development of transplants could be the mononucleate cells from bone marrow. Aims. The purpose of this study was to study the capacity of bone marrow mononucleate cells to survive the transplant process, and to search for a method that enables tracking of these cells in vivo once they have been implanted. Materials and methods. Bone marrow mononucleate cells were extracted from the femur of rats by means of a Ficoll-Hypaque gradient. The cells under study were modified genetically with an adenovirus that expresses the PFV or which are marked with Hoechst dye. The marked cells were implanted in the striatum of rats with lesions caused by quinolinic acid. Results. The viability of the genetically modified cells was low, whereas that of the cells marked with Hoechst dye was above 90%. The implanted cells survived the transplant at least a month and dispersed away from the site of entry towards the corpus callosum and cortex. Conclusions. We consider that the use of Hoechst dye offers more advantages for tracking these cells in vivo. Mononucleate cells have a number of characteristics that allow them to be included as candidate sources of cells for the treatment of neurodegenerative diseases

[Survival of dopaminergic neurons that were hibernated in vitro for seven days]. / Supervivencia in vitro de neuronas dopaminérgicas hibernadas durante siete días.

INTRODUCTION: The use of fresh foetal tissue in neurotransplants entails considerable problems of logistics that limit its clinical applicability, something that can be resolved by the development of optimal tissue storage procedures that do not affect in vivo viability and survival of dopamine. AIMS. To determine whether 7 days' hibernation affects the survival of mesencephalic tissue in vitro, and to compare it to fresh tissue. MATERIALS AND METHODS: The midbrains of rats were hibernated for 1, 3, 5 and 7 days at 4 degrees C. A cellular suspension was prepared for culture throughout a 7-day period. The number of TH+ cells present in the fresh and hibernated cultures was determined. RESULTS: The morphology of the hibernated and cultured dopaminergic neurons was very similar to that of the fresh cells. Comparing the viability of the hibernated and fresh cells did not reveal any significant differences. No significant differences between the numbers of TH+ neurons were observed at any of the hibernation times. The lowest rate of TH+ cell survival was reached at seven days' hibernation. Significant differences (p < 0.05) were found between the number of TH+ neurons for fresh and hibernated tissue. CONCLUSIONS: Hibernation at 4 degrees C for up to five days guarantees the survival of TH+ cells in vitro, but it is affected by longer times. This procedure could be considered useful for preserving human tissue in clinical transplant applications. These results refer to in vitro conditions; therefore, studies must be conducted to investigate the survival and functionality of hibernated and transplanted neurons in animal models to enable us to evaluate its applicability in neurorestorative therapy.

[Marrow stromal cells cultured in N2-supplemented medium: implications on the generation of neural cells]. / Cultivo de las células estromales de la médula ósea en un medio suplementado con los nutrientes N2: implicaciones sobre la generación de células neurales.

INTRODUCTION: Most of the culture system for in vitro maintenance and neural differentiation of marrow stromal cells (MSCs) use synthetic media supplemented with 10 or 20% fetal bovine serum (FBS). Serum, however, is comprised of unknown quantities of undefined substances which could interfere the effect of exogenous substances on neural differentiation of MSCs. AIM. Here we describe survival of MSCs cultured in culture conditions where serum was reduced at 0.5 and 1% using Bottenstein and Sato's N2 formula (1979) and poly-L-lysine (PLL)-coated substrate. MATERIALS AND METHODS: Stromal cells isolated from rat femurs were cultivated in Dulbecco's modified Eagle medium at 10, 1, 0.5% FBS or in serum free medium containing N2 formula. In serum free medium or at low serum concentration culture surface was coated with PLL. Cell survival was determined by MTT method or by counting viable cells. RESULTS: Survival of MSCs cultured in N2 supplement was reduced at about 40% of that observed in 10% FBS containing medium. Under these conditions cell morphology was also affected. When N2 containing medium was supplemented with FBS at 0.5 or 1% a significant increase of survival with respect to that observed in N2-supplemented cultures was observed. Cells seeded on PLL-coated surface increased their survival by contrast with their homologous cultures seeded on uncoated surface. CONCLUSIONS: The culture system which combines N2 formula with FBS 1% and PLL-coated surface is useful for the maintenance of MSCs. These conditions offer advantages for the study of differentiation of these cells because they reduce the confounding influence of serum. The possible implication of this culture system for the study of neural differentiation by these cells is discussed.

Supervivencia in vitro de neuronas dopaminèrgicas hibernadas durante siete dias / s. t

El uso de tejido fetal fresco en el neurotrasplante presenta considerables dificultades logìsticas que limitan su aplicabilidad clìnica . Este aspecto podrìa solucionarse con el desarrollo de procedimientos optimos de almacenamiento del tejido que no afecten a la viabilidad y la supervivencia dopaminèrgica in vivo. Es objetivo del trabajao determinar si la hibernaciòn durante siete dìas influye sobre la supervivencia del tejido mesencefàlico in vitro y comparar el tejido hibernado con el fresco...(AU)

Supervivencia in vitro de neuronas dopaminérgicas hibernadas durante siete días / Survival of dopaminergic neurons that were hibernated in vitro for seven days

Introducción. El uso de tejido fetal fresco en el neurotrasplante presenta considerables dificultades logísticas que limitan su aplicabilidad clínica. Este aspecto podría solucionarse con el desarrollo de procedimientos óptimos de almacenamiento del tejido que no afecten a la viabilidad y la supervivencia dopaminérgica in vivo. Objetivo. Determinar si la hibernación durante siete días influye sobre la supervivencia del tejido mesencefálico in vitro, y comparar el tejido hibernado con el fresco. Material y métodos. El mesencéfalo de rata se hibernó 1, 3, 5 y 7 días a 4 °C. Se preparó la suspensión celular para cultivar durante siete días. Se determinó el número de células TH+ presentes en los cultivos frescos e hibernados. Resultados. La morfología de las neuronas dopaminérgicas hibernadas y cultivadas fue muy similar a la de las células frescas. La comparación de la viabilidad de las células hibernadas con la de las frescas mostró diferencias no significativas. No hay diferencias significativas entre el número de neuronas TH+ observadas en todos los tiempos de hibernación. La supervivencia de células TH+ más baja se alcanzó a los siete días de hibernación. Existen diferencias significativas (p < 0,05) entre el número de neuronas TH+ del tejido fresco y el hibernado. Conclusiones. La hibernación a 4 °C hasta cinco días garantiza la supervivencia in vitro de las células TH+; tiempos mayores, la afecta. Este procedimiento podría considerarse útil para la conservación del tejido humano aplicable en el trasplante clínico. Estos resultados se refieren a condiciones in vitro; por tanto, se requiere estudiar la sobrevivencia y funcionalidad de las neuronas hibernadas y trasplantadas en modelos animales para evaluar su aplicación en la terapia neurorrestaurativa

Introduction. The use of fresh foetal tissue in neurotransplants entails considerable problems of logistics that limit its clinical applicability, something that can be resolved by the development of optimal tissue storage procedures that do not affect in vivo viability and survival of dopamine. Aims. To determine whether 7 days’ hibernation affects the survival of mesencephalic tissue in vitro, and to compare it to fresh tissue. Materials and methods. The midbrains of rats were hibernated for 1, 3, 5 and 7 days at 4 °C. A cellular suspension was prepared for culture throughout a 7-day period. The number of TH+ cells present in the fresh and hibernated cultures was determined. Results. The morphology of the hibernated and cultured dopaminergic neurons was very similar to that of the fresh cells. Comparing the viability of the hibernated and fresh cells did not reveal any significant differences. No significant differences between the numbers of TH+ neurons were observed at any of the hibernation times. The lowest rate of TH+ cell survival was reached at seven days’ hibernation. Significant differences (p < 0.05) were found between the number of TH+ neurons for fresh and hibernated tissue. Conclusions. Hibernation at 4 °C for up to five days guarantees the survival of TH+ cells in vitro, but it is affected by longer times. This procedure could be considered useful for preserving human tissue in clinical transplant applications. These results refer to in vitro conditions; therefore, studies must be conducted to investigate the survival and functionality of hibernated and transplanted neurons in animal models to enable us to evaluate its applicability in neurorestorative therapy

Cultivo de las celulas estromales de la mèdula òsea en un medio suplementado con los nutrientes N2: implicaciones sobre la generaciòn de cèlulas neurales / s. t

Most of the culture system for in vitro maintenance and neural differentiation of marrow stromal cells (MSCs) use synthetic media supplemented with 10 or 20 percent fetal bovine serum (FBS). Serum, however, is comprised of unknown quantities of undefined substances which could interfere the effect of exogenous substances on neural differentiation of MSCs, AIM. Here we describe survival of MSCs cultured in culture conditions where serum was reduced at 0,5 and 1 percent using Bottenstein and Sato's N2 formula (1979) and poly-L-lysine (PLL)-coated substrate...(AU)

Cultivo de las células estromales de la médula ósea en un medio suplementado con los nutrientes N2: implicaciones sobre la generación de células neurales / Marrow stromal cells cultured in n2-supplemented medium: implications on the generation of neural cells

Introducción. La mayoría de los sistemas de cultivo para el mantenimiento in vitro y la diferenciación neural de las células estromales de la médula ósea (CEM) utilizan medios sintéticos suplementados con suero fetal bovino (SFB) al 10 o al 20%. Sin embargo, el suero se compone de cantidades desconocidas de sustancias no definidas que podrían interferir el efecto de las sustancias exógenas sobre la diferenciación neural de estas células. Objetivo. En este trabajo describimos la supervivencia de las CEM en condiciones de cultivo donde se redujo la concentración del SFB al 0,5 y al 1% y se utilizó la fórmula N2 de Bottenstein y Sato (1979) y un sustrato tratado con poli-L-lisina (PLL). Materiales y métodos. Se cultivaron células estromales aisladas de los fémures de rata en el medio de Eagle modificado por Dulbecco, con concentraciones de SFB del 10, el 1 y el 0,5% o en medio libre de suero, que contenían la fórmula N2. En los cultivos crecidos en medios libres de suero o con baja concentración de éste, la superficie de cultivo se trató con PLL. La supervivencia celular se midió por el método del MTT o por recuento de las células vivas. Resultados. La supervivencia de las CEM cultivadas en la fórmula N2 disminuyó hasta aproximadamente el 40% de la observada en el medio con SFB al 10%, y se afectó la morfología celular. Un aumento significativo de la supervivencia con respecto al cultivo en N2 se produjo cuando, además de este nutriente, se añadió SFB al 0,5 y al 1%. En los cultivos sembrados sobre superficies tratadas con PLL la supervivencia celular aumentó en comparación con los sembrados sobre superficies no tratadas. Conclusiones. Este sistema de cultivo que combina la fórmula N2 con SFB al 1% y emplea un sustrato tratado con PLL, es adecuado para el mantenimiento de las CEM. Estas condiciones son ventajosas para estudiar la diferenciación neural de estas células, ya que reducen la interferencia del suero. Se discute la posible implicación de este sistema de cultivo para los estudios de diferenciación neural en estas células

Introduction. Most of the culture system for in vitro maintenance and neural differentiation of marrow stromal cells (MSCs) use synthetic media supplemented with 10 or 20% fetal bovine serum (FBS). Serum, however, is comprised of unknown quantities of undefined substances which could interfere the effect of exogenous substances on neural differentiation of MSCs. Aim. Here we describe survival of MSCs cultured in culture conditions where serum was reduced at 0.5 and 1% using Bottenstein and Sato’s N2 formula (1979) and poly-L-lysine (PLL)-coated substrate. Materials and methods. Stromal cells isolated from rat femurs were cultivated in Dulbecco’s modified Eagle medium at 10, 1, 0.5% FBS or in serum free medium containing N2 formula. In serum free medium or at low serum concentration culture surface was coated with PLL. Cell survival was determined by MTT method or by counting viable cells. Results. Survival of MSCs cultured in N2 supplement was reduced at about 40% of that observed in 10% FBS containing medium. Under these conditions cell morphology was also affected. When N2 containing medium was supplemented with FBS at 0.5 or 1% a significant increase of survival with respect to that observed in N2-supplemented cultures was observed. Cells seeded on PLL-coated surface increased their survival by contrast with their homologous cultures seeded on uncoated surface. Conclusions. The culture system which combines N2 formula with FBS 1% and PLL-coated surface is useful for the maintenance of MSCs. These conditions offer advantages for the study of differentiation of these cells because they reduce the confounding influence of serum. The possible implication of this culture system for the study of neural differentiation by these cells is discussed

[Stromal cell transplant in the 6-OHDA lesion model]. / Trasplante de células estromales en el modelo de lesión por 6-OHDA.

INTRODUCTION: A good deal of evidence currently exists to show that transplanting foetal mesencephalic tissue can produce symptomatic benefits both in patients and in disease models. Nevertheless, the technical and ethical difficulties involved in obtaining enough suitable foetal cerebral tissue have been a serious obstacle to its application. Stromal cells derived from bone marrow, due to their potential capacity to generate different types of cells, could be an ideal source of material for cell restoration in neurodegenerative diseases. AIMS: Our aim was to evaluate the effect of transplanting stromal cells derived from bone marrow on the behaviour of 6-OHDA rats, when they are inserted into the striatum. MATERIAL AND METHODS: In this study we used rats with a lesion in the substantia nigra induced by 6-hydroxydopamine, divided into several experimental groups. Rotary activity induced by D-amphetamine (5 mg/kg, intraperitoneally) was evaluated before and throughout the three months following the transplant in all the experimental groups, except in the group of healthy controls. Hemiparkinsonian rats received a total of 350 000 foetal ventral mesencephalic cells and 8 x 10(4) stromal cells/microL, which were implanted in the striatum. RESULTS AND CONCLUSIONS: Animals with stromal cells transplanted in the body of the striatum significantly reduced the number of turns induced by amphetamine (p < 0.05); yet this reduction was not greater than that induced by foetal mesencephalic cell transplants. We were also unable to demonstrate any significant improvement in the motor skills of the forelimbs.

Trasplante de células estromales en el modelo de lesión por 6-OHDA / Stromal cell transplant in the 6-OHDA lesion model

A good deal of evidence currently exists to show that transplanting foetal mesencephalic tissue can produce symptomatic benefits both in patients and in disease models. Nevertheless, the technical and ethical difficulties involved in obtaining enough suitable foetal cerebral tissue have been a serious obstacle to its application. Stromal cells derived from bone marrow, due to their potential capacity to generate different types of cells, could be an ideal source of material for cell restoration in neurodegenerative diseases. AIMS: Our aim was to evaluate the effect of transplanting stromal cells derived from bone marrow on the behaviour of 6-OHDA rats, when they are inserted into the striatum(AU)

Trasplante de células estromales en el modelo de lesión por 6-OHDA / Stromal cell transplant in the 6-OHDA lesion model

Introducción. En la actualidad, existe un cúmulo de evidencias de que el trasplante de tejido mesencefálico fetal puede producir un beneficio sintomático tanto en los pacientes con enfermedad de Parkinson como en los modelos de la enfermedad. Sin embargo, las dificultades técnicas y éticas en la obtención de tejido cerebral fetal apropiado y en cantidad suficiente ha dificultado su aplicación. Las células estromales derivadas de médula ósea, debido a su potencialidad para generar diferentes tipos de células, podrían ser una fuente ideal para la restauración celular en las enfermedades neurodegenerativas. Objetivo. Evaluar el efecto del trasplante de células estromales derivadas de médula ósea sobre la conducta de ratas con lesión por 6-OHDA, cuando se realiza en el estriado. Materiales y métodos. Se utilizaron ratas con lesión de la sustancia negra inducida por la 6-OHDA, divididas en varios grupos experimentales. La actividad rotatoria inducida por D-anfetamina (5 mg/kg intraperitonialmente) se evaluó antes y en los tres meses posteriores al trasplante en todos los grupos experimentales, excepto en el grupo de controles sanas. Las ratas hemiparkinsonianas recibieron un total de 350.000 células de mesencéfalo ventral fetal y 8 × 104 células estromales/µL, las cuales se implantaron en el estriado. Resultados y conclusiones. Los animales con trasplante de células estromales en el cuerpo estriado redujeron significativamente el número de vueltas inducidas por anfetamina (p < 0,05); sin embargo, esta reducción no fue mayor que la inducida por los trasplantes de células mesencefálicas fetales. Por otro lado, no fue posible demostrar una mejoría significativa de las habilidades motoras de las extremidades anteriores (AU)

Introduction. A good deal of evidence currently exists to show that transplanting foetal mesencephalic tissue can produce symptomatic benefits both in patients and in disease models. Nevertheless, the technical and ethical difficulties involved in obtaining enough suitable foetal cerebral tissue have been a serious obstacle to its application. Stromal cells derived from bone marrow, due to their potential capacity to generate different types of cells, could be an ideal source of material for cell restoration in neurodegenerative diseases. Aims. Our aim was to evaluate the effect of transplanting stromal cells derived from bone marrow on the behaviour of 6-OHDA rats, when they are inserted into the striatum. Materials and methods. In this study we used rats with a lesion in the substantia nigra induced by 6-hydroxydopamine, divided into several experimental groups. Rotary activity induced by D-amphetamine (5 mg/kg, intraperitoneally) was evaluated before and throughout the three months following the transplant in all the experimental groups, except in the group of healthy controls. Hemiparkinsonian rats received a total of 350,000 foetal ventral mesencephalic cells and 8 × 104 stromal cells/µL, which were implanted in the striatum. Results and conclusions. Animals with stromal cells transplanted in the body of the striatum significantly reduced the number of turns induced by amphetamine (p < 0.05); yet this reduction was not greater than that induced by foetal mesencephalic cell transplants. We were also unable to demonstrate any significant improvement in the motor skills of the forelimbs (AU)

[In vitro viability and glutathione levels in mesencephalic neurons after seven days hibernation]. / Viabilidad in vitro y niveles de glutati n en neuronas mesencef licas hibernadas durante siete d as.

In embryonic mesencephalic transplant in patients with Parkinson s disease dopaminergic survival is low (5 10%), and for this reason the use of multiple donors has been considered. The difficulty of obtaining more tissue determines the need for a procedure that enables human nigral tissue to be stored for a time without affecting its physiological state in any significant way. This study was designed to determine whether hibernation of tissue fragments has any influence on viability, how the viability of the mesencephalic cells behaves after 7 days hibernation and the glutathione levels in the hibernated tissue (HT). The viability of the HT in pieces (82.37 2.12) was found to be higher than the value for the whole mesencephalon (70.29 3.43). Viability of the HT, seven days at 4 C, at different post dissociation times, did not differ significantly. Despite the significant differences found between hibernated and fresh tissue at t= 0, this procedure does not seem to affect the mesencephalic tissue in any significant way, as it conserved a 94% viability after hibernation. No evidence was found of increased glutathione content as an antioxidizing response to the damage that might be caused by hibernation. These results suggest that since hibernation does not have any significant effect on the state of the cells it could be considered a useful procedure for conserving tissue to be used in clinical transplants. Moreover, further research is needed on survival and functionality of hibernated cells after being transplanted into animal models in order to evaluate their potential for use in cell therapy.