Análisis funcional del promotor del gen pitx3 mediante la expresión de un gen reportero en neuronas dopaminérgicas mesencefálicas

El
 aumento
 del
 promedio
 de
 vida
 del
 hombre
 ha
 conducido
 a
 un
 incremento
 en
 el
 número
de
individuos
con
enfermedades
degenerativas,
siendo
el
sistema
nervioso
uno
de
los
 más
 susceptibles
 a
 dañarse
 conforme
 avanza
 la
 edad
 del
 individuo.
 La
 región
 del
 sistema nervioso
 más
 susceptible
 a
 sufrir
 muerte
 neuronal,
 es
 la
 región
 ventral
 del
 mesencéfalo
 o
 cerebro
medio.
Las
neuronas
dopaminérgicas
mesencefálicas
(NDM)
se
encuentran
agrupadas
 en
dos
núcleos
principalmente,
el
Área
Ventral
Tegmental
(VTA)
y
la
Sustancia Negra
(SN).
Las
 NDM
 controlan
 el
 movimiento
 motor
 voluntario,
 las
 actividades
 cognitivas
 y
 de
 conducta
 al
 producir
 el
 neurotransmisor
 dopamina
 y
 liberarlo
 al
 cuerpo
 estriado.
Al
morir
 las
NDM,
 ésta
 comunicación
 se
 pierde
 y
 se
 presentan
 los
 síntomas
 característicos
 de
 la
 Enfermedad
 de
 Parkinson
(EP)

Animal Model of Parkinson Disease: Neuroinflammation and Apoptosis in the 6-Hydroxydopamine-Induced Model

6-Hydroxydopamine (6-OHDA), a synthetic neurotoxin, has been used to generate animal models of Parkinson’s disease (PD). Even though 6-OHDA induced neurodegenerative model in rat, it does not reproduce all the symptoms of the disease, but it does replicate most of the cellular processes such as oxidative stress, neurodegeneration, neuroinflammation and apoptotic neuronal death. The knowledge of the mechanisms involved in neurodegeneration is relevant to define possible therapeutic targets for PD

Estrés oxidativo y capacidad antioxidante en deportistas con dieta rica en antioxidantes con zarzamora (Rubus sp.)

Objetivo. Identificar el comportamiento del Estrés Oxidativo (EO) en atletas con y sin la ingesta de dieta rica en antioxidantes durante el periodo competitivo. Métodos. Se evaluaron a 14 atletas de alto rendimiento del equipo de balonmano, quienes fueron distribuidos 7 en un grupo experimental y 7 en un grupo control (con y sin ingesta de zarzamora, respectivamente). El consumo de la bebida de zarzamora o placebo para ambos grupos, fue de una dosis diaria durante 15 días (7 días en la etapa de pre competencia, 7 días durante el periodo de competencia y una dosis 24h después de finalizar la competencia). Se cuantificó el EO (i.e. prueba d-ROMs, unidades Cornelli, U. Cor.) y la capacidad total antioxidante (CTA) en plasma (i.e. prueba PAT, unidades Carratelli, U. Carr.), en 4 momentos: (1) reposo (1 semana previa a competencia, antes del suministro de bebida); (2) pre competencia (una semana antes de la competencia); (3) al final de competencia y (4) a las 24 h después de la competencia. Resultados. En el grupo experimental, el EO disminuyó de manera significativa (p = .018) al comparar la toma en reposo con la toma previa a la competencia después de 7 días de la ingesta de la dieta rica en antioxidantes. El grupo control presentó aumentos significativos de la CTA en la toma previa a competencia (p = .028) así como al final de la misma (p = .046), con respecto a la toma en reposo. Conclusión. El EO se incrementa después de la competencia y estimula la CTA. La ingesta de la dieta rica en antioxidantes es favorable en el entrenamiento previo a la competencia ya que promueve la regulación del EO, disminuyendo los valores del mismo.Objective. Identify Oxidative Stress (OS) behavior in athletes with and without the intake of an antioxidant-rich diet during a competitive period. Methods.14 high-performance athletes of handball team were evaluated. Two groups were established: 7 in an experimental group (blackberry intake) and 7 in a control group (with and without the intake of blackberry, respectively). The intake of blackberry beverage or placebo for both groups was a daily dose for 15 days (7 days in the pre-competition stage, 7 days during the competition period and a dose 24h after the end of the competition). OS (i.e. d-ROMs test, Cornelli units, U.Cor.) and the Total Antioxidant Capacity (TAC) (i.e. PAT test, Carratelli units, U. Carr.) on plasma, were quantified at 4 moments: (1) resting (1 week before the competition, before the beverage intake); (2) pre competition (one week before the competition); (3) at the end of the competition; and (4) 24 hours after the competition. Results. On the experimental group, OS was significantly reduced (p =.018) comparing resting takes with the pre-competition, after 7 days of the antioxidant-rich diet. The control group had a significant rise on TAC presented in precompetition (p =.028) as well as at end of competition (p =.046) compared to the resting take. Conclusion. The OS rises after competition and stimulates the TAC. The intake of an antioxidant-rich diet is helpful on pre competition training since it promotes the regulation of OS, diminishing its levels.Objetivo. Identificar o comportamento do estresse oxidativo (EO) em atletas com e sem a ingestão de dieta rica em antioxidantes durante o período competitivo. Métodos. Foram avaliadas 14 atletas de alto rendimento da equipe de handebol, que foram distribuídos em 7 num grupo experimental e 7 num grupo de controlo (com e sem ingestão de amora, respectivamente). O consumo da amora bebida ou placebo em ambos os grupos era uma dose diária durante 15 dias (7 dias no pré competição, 7 dias durante a competição e uma dose 24 horas depois de terminar a competição). O EO foi quantificado (i.e. prova d-ROMs, unidades Cornelli, U. Cor.) e a capacidade antioxidante total (CAT) no plasma (isto é, teste de PAT, unidades Carratelli, U. Carr.) em 4 fases: (1) repouso (uma semana antes da competição antes do fornecimento de bebida); (2) pré-competição (uma semana antes da competição); (3) no fim da competição e (4) às 24 h após a competição. Resultados. No grupo experimental, o EO diminuiu significativamente (p = 0,018) comparando a toma em repouso com a toma anterior à competição após 7 dias de ingestão da dieta rica em antioxidantes. O grupo de controlo teve aumentos significativos no CAT na toma previa à competição (p = .028) e no final da mesma (p = .046), com respeito à toma em repouso. onclusão. O EO aumenta após da competição e estimula o CAT. A ingestão de dieta rica em antioxidantes é favorável no treinamento pré-competição, uma vez que promove a regulação da EO, diminuindo os valores do mesmo

Cytological effects of pulsed electromagnetic fields and static magnetic fields induced by a therapeutic device on in vivo exposed rats

Background: There is a trend towards the use of magnetic fields in medicine. Pulsed electromagnetic fields (PEMFs) technology was based upon 20 years of fundamental studies on the electromechanical properties of bone and other connective tissues. More recently, these magnetic fields have been used to treat several health conditions. There remains continuing concern that exposure to electromagnetic devices may cause adverse effects. The aim of the present study was to investigate the cytological effects induced in rats exposed in a patented medical device that uses PEMFs combined with static magnetic fields (SMFs).Material and Methods: Thirty sexually mature 14-week-old male and female Sprague Dawley rats were distributed into three groups: (a) 5 males and 5 females (independently) exposed to PEMFs combined with SMFs, (b) animals treated with SMFs only, and (c) non-exposed animals. Acridine orange fluorescent-staining micronucleus test and male germ cells analysis were performed according to standardized techniques.Results: A lack of evidence for alterations on micronucleus frequency, on polychromatic erythrocytes percentage, and on sperm counts and morphological characteristics of male germ cells were found in mature rats exposed to PEMFs medical device compared to non-exposed animals.Conclusions: This study suggests that the applied magnetic field generated in a therapeutic device did not have any detectable cytotoxic or genotoxic effect in exposed rats. In view of these findings and the contradictory reports in the literature, it is necessary to carry out more research to help clarify the controversy concerning cytogenotoxic risk associated with therapeutic magnetic fields exposures.Keywords: Cytotoxicity, pulsed electromagnetic fields, static magnetic fields, micronuclei, sperm abnormalitie

Production of biologically active human lymphotactin (XCL1) by Lactococcus lactis

Lymphotactin-XCL1 is a chemokine produced mainly by activated CD8? T-cells and directs migration of CD4? and CD8? lymphocytes and natural killer (NK) cells. We expressed human lymphotactin (LTN) by the lactic-acid bacterium Lactococcus lactis. Biological activity of LTN was confirmed by chemo-attraction of human T-cells by chemotaxis demonstrating, for the first time, how this chemokine secreted by a food-grade prokaryote retains biological activity and chemoattracts T lymphocytes. This strain thus represents a feasible well-tolerated vector to deliver active LTN at a mucosal level

Antioxidant gene therapy against neuronal cell death

Oxidative stress is a common hallmark of neuronal cell death associated with neurodegenerative disorders such as Alzheimer\u27s disease, Parkinson\u27s disease, as well as brain stroke/ischemia and traumatic brain injury. Increased accumulation of reactive species of both oxygen (ROS) and nitrogen (RNS) has been implicated inmitochondrial dysfunction, energy impairment, alterations in metal homeostasis and accumulation of aggregated proteins observed in neurodegenerative disorders, which lead to the activation/modulation of cell death mechanisms that include apoptotic, necrotic and autophagic pathways. Thus, the design of novel antioxidant strategies to selectively target oxidative stress and redox imbalance might represent important therapeutic approaches against neurological disorders. This work reviews the evidence demonstrating the ability of genetically encoded antioxidant systems to selectively counteract neuronal cell loss in neurodegenerative diseases and ischemic brain damage. Because gene therapy approaches to treat inherited and acquired disorders offer many unique advantages over conventional therapeutic approaches, we discussed basic research/clinical evidence and the potential of virus-mediated gene delivery techniques for antioxidant gene therapy

The Causative and Curative Roles of Brain-Derived Neurotrophic Factor in Parkinson’s Disease

Parkinson’s disease (PD) is characterized by the activation of degenerative and inflammatory processes in brain circuits that control movement and, according to the degree of progression of the damage, can cause neuropsychological disorders such as cognitive dysfunction. Changes in gene expression profile or post-translational modifications in secretory proteins such as neurotrophic factors could define the disease progression. Brain-derived neurotrophic factor (BDNF) is relevant, because it not only participates in neuronal survival, neurotransmission, dendritic growth and cellular communication but also in disease progression. In this chapter, considering both experimental evidences and clinical reports, the authors will analyze the contribution of BDNF as one of the causes of neurodegeneration and neuroinflammation; discuss the participation of this neurotrophic factor in the development of cognitive dysfunction, and finally the scope of novel BDNF-based therapies for PD

Efficient secretion of a modified E7 protein from human papilloma virus type-16 by Lactococcus lactis

Aims: To create and provide a strain of the food-grade bacterium Lactococcus lactis able to efficiently secrete a modified form of the E7 protein from the human papilloma virus (HPV) type-16. Methods and Results: We cloned the coding sequence of a modified E7 (E7m)from the HPV-16 in a plasmid regulated by the strong expression promoter p59. Secretion of the E7m was made by the signal peptide of the usp45 gene.The E7m was detected by Western blot in the cell-free-medium fraction, showing no degradation or aberrant forms. Conclusions: We constructed a strain of L. lactis able to secrete efficiently aHPV-16 E7 modified protein with diminished transforming activity. Significance and Impact of the Study: Human papilloma virus infection is associated with more than 99% of cervical cancers. Immunotherapy targeting E7 to treat HPV-associated cervical malignancies has been demonstrated to be highly efficient. However, native E7 maintains transforming activity. We present this new strain of a food-grade bacterium able to efficiently secrete a HPV-16 E7-modified protein with diminished transforming activity. This new strain could be used as a live vaccine to deliver E7 at a mucosal level and generate antitumour immune responses against HPV-associated tumours