Histological Features of the Olive Seed and Presence of 7S-Type Seed Storage Proteins as Hallmarks of the Olive Fruit Development.

The production of olive oil is an important economic engine in the Mediterranean area. Nowadays, olive oil is obtained mainly by mechanical processes, by using the whole fruit as the primary raw material. Although the mesocarp is the main source of lipids contributing to olive oil formation, the seed also contributes to the olive oil composition and attributes. The olive seed is also becoming an interesting emerging material itself when obtained after alternative processing of the olive fruit. Such seed is used for the production of differential oil and a unique flour among other bioactive products, with increasing uses and applications in cosmetics, nutrition, and health. However, olive seed histology has been poorly studied to date. A complete description of its anatomy is described for the first time in the present study by using the 'Picual' cultivar as a model to study the development of the different tissues of the olive seed from 60 to 210 days after anthesis. A deep analysis of the seed coats, endosperm storage tissue and the embryo during their development has been performed. Moreover, a panel of other olive cultivars has been used to compare the weight contribution of the different tissues to the seed, seed weight variability and the number of seeds per fruit. In addition to the histological features, accumulation of seed storage proteins of the 7S-type (ß-conglutins) in the seed tissues has been assessed by both biochemical and immunocytochemical methods. These hallmarks will help to settle the basis for future studies related to the location of different metabolites along the olive seed and mesocarp development, and therefore helping to assess the appropriate ripening stage for different commercial and industrial purposes.

Melatonin rescues zebrafish embryos from the parkinsonian phenotype restoring the parkin/PINK1/DJ-1/MUL1 network.

Multiple studies reporting mitochondrial impairment in Parkinson's disease (PD) involve knockout or knockdown models to inhibit the expression of mitochondrial-related genes, including parkin, PINK1, and DJ-1 ones. Melatonin has significant neuroprotective properties, which have been related to its ability to boost mitochondrial bioenergetics. The meaning and molecular targets of melatonin in PD are yet unclear. Zebrafish are an outstanding model of PD because they are vertebrates, their dopaminergic system is comparable to the nigrostriatal system of humans, and their brains express the same genes as mammals. The exposure of 24 hpf zebrafish embryos to MPTP leads to a significant inhibition of the mitochondrial complex I and the induction of sncga gene, responsible for enhancing γ-synuclein accumulation, which is related to mitochondrial dysfunction. Moreover, MPTP inhibited the parkin/PINK1/DJ-1 expression, impeding the normal function of the parkin/PINK1/DJ-1/MUL1 network to remove the damaged mitochondria. This situation remains over time, and removing MPTP from the treatment did not stop the neurodegenerative process. On the contrary, mitochondria become worse during the next 2 days without MPTP, and the embryos developed a severe motor impairment that cannot be rescued because the mitochondrial-related gene expression remained inhibited. Melatonin, added together with MPTP or added once MPTP was removed, prevented and recovered, respectively, the parkinsonian phenotype once it was established, restoring gene expression and normal function of the parkin/PINK1/DJ-1/MUL1 loop and also the normal motor activity of the embryos. The results show, for the first time, that melatonin restores brain function in zebrafish suffering with Parkinson-like disease.

The role of nitric oxide during healing of trauma to the skeletal muscle.

INTRODUCTION: The role of NO in muscle injury is not clear. METHODS: We examined the involvement of the NO system in the development of muscle damage in an experimental model of crush injury. The animals were divided into four groups: (1) control (CO), (2) sham trauma, (3) trauma, (4) trauma + L -NAME, in two experimental phases, 24 h and 7 days after injury. RESULTS: Twenty-four hours post-trauma, the crushed muscle was characterized by an intense inflammatory reaction. These changes were accompanied by increased oxidative damage, increased cytokine mRNA transcription, NF-κB binding ability and TGF-ß growth factor expression in the gastrocnemius muscle. Treatment with L: -NAME markedly decreased these histological and molecular abnormalities at 24 h. However, at 7 days post-trauma, increased collagen formation was observed in the L: -NAME group. DISCUSSION: These findings indicate that NO is involved in the balance between fibrosis and healing with regeneration.

Nitric oxide regulates the repair of injured skeletal muscle.

Skeletal muscle repair can be understood as a balance between fibrosis and regeneration, the result of which may lead to complete recovery or loss of muscle function. To study the involvement of nitric oxide in post-trauma muscle repair, we used an experimental murine model of crush injury muscle. The animals were divided into four groups, (i) control (CO), (ii) sham trauma, (iii) trauma and (iv) trauma+l-NAME. The animals received a single dose of 100mg/kg of the l-NAME, an inhibitor of nitric oxide synthase, 2h after lesion, and the muscle tissue was analyzed in two time-points: 24h and 7 days. Twenty-four hours after injury, the crushed muscle was characterized by an intense inflammatory cell infiltrate and edema demonstrated by histological analysis. These changes were accompanied by increased iNOS, MMP-2 and HGF mRNA transcription and protein expression of the iNOS and MMP-2 in the gastrocnemius muscle. Crushing injury also promoted cell proliferation and increase number satellite cell, responsible for the regeneration of the muscle fiber. Treatment with l-NAME blocking local NO production, greatly attenuated these histological and molecular findings at 24h. On the 7th day the molecular findings of both groups were comparable to the control (sham trauma) group. However, the l-NAME group showed increase deposition of collagen and decrease of SC expression. These findings demonstrate that activation of NO during muscle crush is critical in the early phases of the skeletal muscle repair process and indicate its possible role as a regulator of the balance between fibrosis and muscle regeneration.

Eccentric training impairs NF-kappaB activation and over-expression of inflammation-related genes induced by acute eccentric exercise in the elderly.

The present study was aimed to investigate in elderly humans changes in NF-kappaB activation and in the expression of the inflammation-related genes inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and interleukin-6 (IL-6) induced in peripheral blood mononuclear cells (PBMC) by acute eccentric exercise and by submaximal eccentric training. Eleven subjects, aged 66-75 years, carried out 2 bouts of eccentric exercise separated by 8 weeks of training. Following the first bout, NF-kappaB activation, and protein level of p50/p65 subunits, phospho-IkappaBalpha and phospho-IKKalpha increased, while IkappaBalpha protein level was significantly reduced. This was accompanied by a significant increase in iNOS, COX-2 and IL-6 mRNA protein level and protein content. Changes were significantly attenuated following the second exercise bout. In conclusion, acute eccentric exercise increases NF-kappaB activation and the expression of several inflammation-related genes in PBMC from elderly individuals. Regular eccentric training might be an effective method of preventing undesirable inflammatory responses induced by eccentric exercise.

Effects of eccentric exercise on NF-kappaB activation in blood mononuclear cells.

PURPOSE: Changes in nuclear factor kappaB (NF-kappaB) activation induced in peripheral blood mononuclear cells (PBMC) by acute eccentric exercise and by submaximal eccentric training were investigated. METHODS: Eleven subjects carried out two bouts of eccentric exercise separated by 6 wk of training. RESULTS: Soreness, vertical jump height, and plasma creatine kinase were significantly modified after the first bout. NF-kappaB activation, p50 and p65, phospho-IkappaBalpha and phospho-IKK protein level, and Mn-SOD expression increased in PBMC, whereas IkappaBalpha protein level was significantly reduced. Changes were significantly attenuated after the second exercise bout. An additional group of nine subjects carried out the two bouts of exercise without training. Effects on NF-kappaB activation were similar after the second bout compared with the first, despite a reduction in markers of muscle injury (repeated bout effect). CONCLUSION: Training significantly attenuates the NF-kappaB-dependent pathway changes induced in PBMC by eccentric exercise, with no contribution from the repeated bout effect.