Signalling pathways regulating muscle mass in ageing skeletal muscle: the role of the IGF1-Akt-mTOR-FoxO pathway.

During ageing skeletal muscles undergo a process of structural and functional remodelling that leads to sarcopenia, a syndrome characterized by loss of muscle mass and force and a major cause of physical frailty. To determine the causes of sarcopenia and identify potential targets for interventions aimed at mitigating ageing-dependent muscle wasting, we focussed on the main signalling pathway known to control protein turnover in skeletal muscle, consisting of the insulin-like growth factor 1 (IGF1), the kinase Akt and its downstream effectors, the mammalian target of rapamycin (mTOR) and the transcription factor FoxO. Expression analyses at the transcript and protein level, carried out on well-characterized cohorts of young, old sedentary and old active individuals and on mice aged 200, 500 and 800 days, revealed only modest age-related differences in this pathway. Our findings suggest that during ageing there is no downregulation of IGF1/Akt pathway and that sarcopenia is not due to FoxO activation and upregulation of the proteolytic systems. A potentially interesting result was the increased phosphorylation of the ribosomal protein S6, indicative of increased activation of mTOR complex1 (mTORC1), in aged mice. This result may provide the rationale why rapamycin treatment and caloric restriction promote longevity, since both interventions blunt activation of mTORC1; however, this change was not statistically significant in humans. Finally, genetic perturbation of these pathways in old mice aimed at promoting muscle hypertrophy via Akt overexpression or preventing muscle loss through inactivation of the ubiquitin ligase atrogin1 were found to paradoxically cause muscle pathology and reduce lifespan, suggesting that drastic activation of the IGF1-Akt pathway may be counterproductive, and that sarcopenia is accelerated, not delayed, when protein degradation pathways are impaired.

Food restriction reverses the hyper-muscular phenotype and force generation capacity deficit of the myostatin null mouse.

Food restriction has a great impact on skeletal muscle mass by inducing muscle protein breakdown to provide substrates for energy production through gluconeogenesis. Genetic models of hyper-muscularity interfere with the normal balance between protein synthesis and breakdown which eventually results in extreme muscle growth. Mutations or deletions in the myostatin gene result in extreme muscle mass. Here we evaluated the impact of food restriction for a period of 5 weeks on skeletal muscle size (i. e., fibre cross-sectional area), fibre type composition and contractile properties (i. e., tetanic and specific force) in myostatin null mice. We found that this hyper-muscular model was more susceptible to catabolic processes than wild type mice. The mechanism of skeletal muscle mass loss was examined and our data shows that the myostatin null mice placed on a low calorie diet maintained the activity of molecules involved in protein synthesis and did not up-regulate the expression of genes pivotal in ubiquitin-mediated protein degradation. However, we did find an increase in the expression of genes associated with autophagy. Surprisingly, the reduction on muscle size was followed by improved tetanic and specific force in the null mice compared to wild type mice. These data provide evidence that food restriction may revert the hyper-muscular phenotype of the myostatin null mouse restoring muscle function.

An additional performance of HTRS: the waste radiotoxicity minimisation.

The management of radioactive waste is a key issue for the present and future use of nuclear energy. In this frame, high temperature reactors (HTRs) have, among others, the capability to burn actinides. After a short introduction on HTRs, the performances of two MC-based burnup codes (Monte Carlo continuous energy burnup and MONTEBURNS) in assessing the ability of these reactors to burn actinides are compared. These codes are necessary for performing ultra-high burnup calculations on HTRs. The best one, in this specific case, results to be MONTEBURNS. It was analysed using HTRs loaded with the following: (1) 1st generation Pu, 600 equivalent full power days; (2) 2nd generation Pu, 645 equivalent full power days; and (iii) 33% 1st generation Pu and 67% Th, 705 equivalent full power days. Finally, it is possible to conclude that HTRs can reduce time when the waste is considered dangerous. Even if the amount of reduction does not solve the whole problem, it represents an important step in the management of radioactive waste.

Lifelong physical exercise delays age-associated skeletal muscle decline.

Aging is usually accompanied by a significant reduction in muscle mass and force. To determine the relative contribution of inactivity and aging per se to this decay, we compared muscle function and structure in (a) male participants belonging to a group of well-trained seniors (average of 70 years) who exercised regularly in their previous 30 years and (b) age-matched healthy sedentary seniors with (c) active young men (average of 27 years). The results collected show that relative to their sedentary cohorts, muscle from senior sportsmen have: (a) greater maximal isometric force and function, (b) better preserved fiber morphology and ultrastructure of intracellular organelles involved in Ca(2+) handling and ATP production, (c) preserved muscle fibers size resulting from fiber rescue by reinnervation, and (d) lowered expression of genes related to autophagy and reactive oxygen species detoxification. All together, our results indicate that: (a) skeletal muscle of senior sportsmen is actually more similar to that of adults than to that of age-matched sedentaries and (b) signaling pathways controlling muscle mass and metabolism are differently modulated in senior sportsmen to guarantee maintenance of skeletal muscle structure, function, bioenergetic characteristics, and phenotype. Thus, regular physical activity is a good strategy to attenuate age-related general decay of muscle structure and function (ClinicalTrials.gov: NCT01679977).

Investigating incorporation and distribution of radionuclides in trinitite.

Most of the surface explosions in nuclear tests have released radioactivity to the environment in the form of bulk glassy materials originating from the melting of sandy soil in the neighbourhood of ground zero. In view of clarifying issues concerning the mechanism of formation and the radiological impact of these materials, we investigated incorporation and volume distribution of radionuclides in a typical fragment of trinitite, the glassy substance generated following the first nuclear test (Trinity Site, New Mexico, 1945). Specific activities were determined by γ-spectrometry for the most significant fission and activation products. In particular, (152)Eu activity was used to estimate the original point of collection of the sample with respect to ground zero. After embedding in an epoxy resin, the sample was then sliced to perform cross-sectional ß- and α-autoradiograph. α-spectrometry was also carried out on a fine powder obtained by surface abrasion. In the ß-autoradiography, hot spots were distinguishable in the proximity of the blast side, over a 1000 times less intense background of sand activation products. Also α-contamination (from (239+240)Pu and (241)Am) was mostly concentrated within the superficial layer, in a fraction of only 20% of the overall volume of the sample, exhibiting a discontinuous, droplet-like distribution. This evidence would partially support a recent hypothesis on trinitite formation according to which most of the glass layer was formed not on the ground but by a rain of material injected into the fireball that melted, fell back, and collected on a bed of already fused sand.

Cloning, expression, purification, and characterization of Streptococcus pneumoniae IgA1 protease.

The IgA1 protease of Streptococcus pneumoniae is a Zn-metalloproteinase of 1964 amino acids that specifically cleaves the hinge region of IgA1, the predominant class of immunoglobulin present on mucosal membranes. This protease is associated to the bacterial cell surface via an N-terminal membrane anchor. Following proteolysis it is released in several forms of different molecular weight. Here, we describe the cloning, expression, and characterization of the enzymatic activity and immunogenicity of three fragments of IgA1 protease, including a large one lacking only the 103 N-terminal amino acids that constitute a typical prokaryotic signal sequence. Further, a proteolytically inactive mutant was generated by replacement of the glutamate residue with an alanine residue in the active site motif HExxH (1605-1609). This is the first report of recombinant active forms of S. pneumoniae IgA1 protease, which open the possibility of identifying specific inhibitors that could interfere with the mucosal colonization by pneumococcus. Moreover the inactive mutant could be considered as a candidate vaccine component.