Nutrients and pathways that regulate health span and life span

Both life span and health span are influenced by genetic, environmental and lifestyle factors. With the genetic influence on human life span estimated to be about 20\u201325%, epigenetic changes play an important role in modulating individual health status and aging. Thus, a main part of life expectance and healthy aging is determined by dietary habits and nutritional factors. Excessive or restricted food consumption have direct effects on health status. Moreover, some dietary interventions including a reduced intake of dietary calories without malnutrition, or a restriction of specific dietary component may promote health benefits and decrease the incidence of aging-related comorbidities, thus representing intriguing potential approaches to improve healthy aging. However, the relationship between nutrition, health and aging is still not fully understood as well as the mechanisms by which nutrients and nutritional status may affect health span and longevity in model organisms. The broad effect of different nutritional conditions on health span and longevity occurs through multiple mechanisms that involve evolutionary conserved nutrient-sensing pathways in tissues and organs. These pathways interacting each other include the evolutionary conserved key regulators mammalian target of rapamycin, AMP-activated protein kinase, insulin/insulin-like growth factor 1 pathway and sirtuins. In this review we provide a summary of the main molecular mechanisms by which different nutritional conditions, i.e., specific nutrient abundance or restriction, may affect health span and life span

Induced pluripotent stem cell-based models: Are we ready for that heart in a dish?

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Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. For example, a key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process versus those that measure fl ux through the autophagy pathway (i.e., the complete process including the amount and rate of cargo sequestered and degraded). In particular, a block in macroautophagy that results in autophagosome accumulation must be differentiated from stimuli that increase autophagic activity, defi ned as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (inmost higher eukaryotes and some protists such as Dictyostelium ) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the fi eld understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. It is worth emphasizing here that lysosomal digestion is a stage of autophagy and evaluating its competence is a crucial part of the evaluation of autophagic flux, or complete autophagy. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. Along these lines, because of the potential for pleiotropic effects due to blocking autophagy through genetic manipulation it is imperative to delete or knock down more than one autophagy-related gene. In addition, some individual Atg proteins, or groups of proteins, are involved in other cellular pathways so not all Atg proteins can be used as a specific marker for an autophagic process. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

Metabolic Flexibility for Metabolic Health: Role of Nutrition and Nutraceuticals

Both lifespan and health span are influenced by nutrition, with nutritional interventions proving to be robust across a wide range of species. However, mechanisms by which nutrients and nutritional status may affect health span are not fully understood. Both health span and life span are closely linked to metabolic health and this condition mainly depends on metabolic flexibility. Metabolic flexibility is a term coined by Kelley et al. [1], and is conceptually defined as the ability to efficiently adapt metabolism by substrate sensing, trafficking, storage, and utilization, in response to availability and requirement of nutrients as well as by physical activity. In health conditions, metabolic flexibility, i.e. metabolic plasticity, is essential to maintain energy homeostasis. As example, skeletal muscle of lean or physically active individuals showed a remarkable ability to adapt fuel preference to nutrient supply and was therefore designated as metabolically flexible. Upon consumption of a high-fat diet, lean subjects were able to increase Fatty Acid Oxidation (FAO) at the expense of glucose, whereas obese individuals were not [2]. Lean or physically active individuals also showed an increased expression of genes involved in fatty acid transport and oxidation compared with little or no change in their obese counterparts [3]. The ability to shift fuel source oxidation from carbohydrates to fats is generally related to metabolic health. Studies on mice showed that metabolic flexibility correlates with a healthy Respiratory Exchange Ratio (RER) resulting from the circadian shift between carbohydrate (value of 1.0) to lipid metabolism (value of 0.7) [4]. Mitochondrial dysfunction lead to a cellular shift toward a glycolytic phenotype, which is intimately linked, to a sedentary lifestyle and senescence. Metabolic flexibility is negatively correlated with aging and is disrupted in some pathological conditions, as in ectopic lipid accumulation which is causally linked to insulin resistance, in the context of obesity and metabolic syndrome [5]. Insulin-resistant obese patients manifest a lesser reliance on fatty acid oxidation compared with lean individuals and do not show increased fatty acid oxidation after fasting or reduced fatty acid oxidation after insulin infusion. Because of their inadequate responses to metabolic challenges, these patients are named \u201cmetabolically inflexible\u201d [6]. Metabolic inflexibility is a hallmark of many age-related metabolic diseases but also plays a central role in, for instance, cancer and immune metabolism diseases. Conversely, metabolic flexibility is enhanced by lifestyle interventions including exercise training and controlled Calorie Restriction (CR), which are able to reduce obesity, visceral fat deposit and ectopic lipid accumulation [7]. Exercise, in particular, is a principal preventive strategy to improve metabolic flexibility at all ages and prolong healthy aging [8]. These interventions are able to favor mitochondrial function and improve substrate switching and metabolic health. Molecular and signaling pathways drive metabolic flexibility and often serve as metabolic sensors able to respond to different nutritional conditions or exercise. Pathways involved in metabolic flexibility are those mediated by Mammalian Target of Rapamycin (mTOR) and insulin/insulin-like Growth Factor-1 (IGF-1) which are generally stimulated in fed conditions, and by pathways activated by fasting, involving AMP-Activated Protein Kinase (AMPK), NAD+- dependent sirtuin (SIRT) deacetylases and PPAR\u3b3 coactivator 1-a PGC1a) (see ref 11 for a review). Main sensor of low energy levels is AMPK, which mediates one of the \u201cchief\u201d step of metabolic flexibility represented by \u201cglucose-fatty acid cycle\u201d [9]. This step states that high glucose availability suppresses oxidation of fatty acids and vice versa [5]. In particular, during CR, the rise in AMP/ATP activates AMPK, which inhibits Acetyl-Coenzyme a Carboxylase (ACC), thus stimulating fatty acid uptake by the mitochondria via Carnitine- Palmitoyl Transferase 1 (CPT-1) and increasing FAO. In parallel, during CR, the increase of NAD+ concentrations stimulates nuclear/ cytoplasmic-localized SIRT1 and mitochondrial SIRT3 activity and leads to protein deacetylation and improved mitochondrial function. There is a reciprocal interplay between AMPK and SIRT, which contributes to metabolic adaptations during fasting conditions [10] as well as during aerobic exercise. This interplay leads to increased transcription, translation, and activity of the transcriptional coactivator PGC1a which is a main mediator of mitochondrial biogenesis and regulator of exercise-induced adaptations in the capacity of oxidative phosphorylation in skeletal muscle [11,12]. In the last years many food-derived natural compounds, also named nutraceuticals have been investigated in relation to their effects on most of the nutrient sensing pathways activated by CR and physical exercise, and potentially related to longevity and health span [13- 15]. However, in our opinion there are only few controlled studies on the effects of single dietary components on biochemical pathways and enzymes able to improve metabolic flexibility. As example, one relevant aspect might be related to prevention of ectopic intracellular lipid accumulation. At this purpose biochemical studies on nutritional modulation of enzymes involved in substrate switching such as Pyruvate Dehydrogenase (PDH) and PDH kinase 4 (PDK4) as well as in pathways leading to cytosolic acetyl-coenzyme A (acetyl-CoA) accumulation could be of great interest. Nucleo-cytosolic acetyl-CoA has emerged as a central signaling node used to coordinate metabolic flexibility in response to a changing of nutritional status. In fact, cells utilize acetyl-CoA levels to integrate nutrient status with energy levels to ensure the proper funneling of substrate toward energy production or storage. In cytosol acetyl-CoA is generated by the enzyme ATP Citrate Lyase (ACLY) which catalyzes the cleavage of citrate to oxaloacetate and acetyl-CoA, a critical reaction linking cellular glucose metabolism and lipogenesis. Accumulation of acetyl-CoA in cytosol also favors protein acetylation and inhibits autophagy with a negative impact for metabolic health. Recently the involvement of ACLY in the progression and development of various chronic diseases has been comprehensively described [16]. Preclinical studies and clinical randomized trials showed the importance of ACLY activity in metabolism, supporting its inhibition as a potential therapeutic approach to treat atherosclerotic cardiovascular disease, nonalcoholic fatty liver disease and other metabolic disorders [17]. Among nutraceuticals Garcinia cambogia, which contains Hydroxycitric Acid (HCA) has been reported to play a role in inhibiting the enzyme ACLY [14]. However, the safety of this plant extract has been highly questioned [18]. In our opinion further studies are needed that address more exhaustive role of nutrition and nutraceuticals on pathways involved in regulation of metabolic flexibility as well as on any other process affecting cellular metabolic health

Molecular mechanisms linking nutrition to metabolic homeostasis: An overview picture of current understanding

Increasing evidence supports the notion that in humans many pathological conditions including obesity, metabolic syndrome, and type 2 diabetes are closely related to the amount and quality of each nutritional component and to an impairment of the metabolic homeostatic mechanisms of their utilization. Cell signaling pathways that sense the availability of nutrients and the energy status of the cells communicate with signaling pahways triggered by hormones and growth factors to coordinately regulate whole-body metabolic homeostasis. The aim of this review is to provide an overview picture of current knowledge about the main molecular mechanisms that connect nutritional status, hormones, and nutrient levels with gene expression, metabolic homeostasis, and nutrient sensing. We recapitulate molecular mechanisms governing fuel selection between glucose and fatty acids in different nutritional conditions, highlighting metabolic flexibility as mechanism to ensure metabolic health. Disrupted metabolic flexibility, or metabolic inflexibility, is associated with many pathological conditions including metabolic syndrome, type 2 diabetes mellitus, and cancer. We also describe how macronutrients that can be used as energy sources may reciprocally modulate their own metabolism as well as directly interact with transcriptional factors, nutrient sensors and nutrient sensing pathways in order to achieve metabolic homeostasis

Oral health care in patient with Williams Syndrome

William Syndrome is characterized by multiple anomalies including peculiar facial and oral features. A precocious maxillofacial and dental examination is mandatory in order to follow facial development and tooth eruption. The clinical examination often reveals dental anomalies in the form of amelogenesis imperfecta, morphology (microdontia) and number anomalies (dental agenesis). The risk of Early Childhood Caries is increased by gastroesophageous reflux, a frequent finding in the first year of life of this children. Orthopaedic-orthodontic treatment is often required to address bone growth and to correct the position of the teeth both for function and for aesthetic reasons. The cooperation level and the presence of systemic diseases have to be considered before the treatment planning. A correct psychological approach can ameliorate the compliance even in the patient with mental delay, but a certain degree of collaboration is required for positioning orthodontic devices. A careful analysis of the general health of the patient is necessary before dental therapy: antimicrobial prophylaxis, to prevent infective endocarditis is needed in patient with congenital heart diseases (75%); renal function and blood pressure have to be monitorized before administrating drugs (antibiotics, antiflogistics and sedative). A strict collaboration with the paediatrist and other specialists is needed to plan a safe and successful therapy. Parents must be involved in the planning of dental treatment and correctly informed and motivated about oral hygiene technique, as their collaboration is necessary to maintain oral health of their child after the treatment. In this presentation, the authors report their experience in dental treatment in children with Williams Syndrome describing their clinical approach from a complete anamnesis to the treatment plan, in order to propose guidelines for oral health care in this patients. References: Axelsson S., Bjornland T., Kjaer I., Heidberg A., Storhaug K., Dental characteristics in Williams syndrome: a clinical and radiographic evaluation, Acta Odontol Scand. 2003 Jun;61(3):129-36 Hertzberg J, Nakisbendi L, Needleman HL, Pober B Williams syndrome-oral presentation of 45 cases Pediatr Dent. 1994 Jul-Aug;16(4):262-7 Mass E, Belostoky L., Craniofacial morphology of children with Williams syndrome, Cleft Palate Craniofac J, 1993 May;30(3):343-9. Oncag V., Gumbay S., Parlar A., Williams Syndrome, J Clin Ped Dent, 1995; 19(4): 301-

Pedagogia delle immagini e odontoiatria nei bambini con Disturbi Pervasivi dello Sviluppo

Scopo dello studio: è stato quello di verificare l’utilità del metodo educativo “per immagini” TEACH (Treatment and Education of Autistic and related Communication handicapped Children) in soggetti con disturbi pervasivi dello sviluppo (DPS), per facilitare il loro primo avvicinamento allo studio odontoiatrico. Materiali e metodi: sono stati scelti 2 gruppi: test e controllo. Entrambi i gruppi comprendono 11 pazienti con DPS tra i 5 ed i 14 anni con scarsa collaborazione alla prima visita. Il gruppo test è stato sottoposto al metodo TEACH. E’ stato quindi preparato per il gruppo test uno speciale album fotografico contenente le fotografie degli operatori e dell’ambulatorio odontoiatrico e una semplice descrizione delle stesse immagini. Una copia di tale album è stata inoltre consegnata ai genitori e/o ai riabilitatori almeno 15 giorni prima dell’appuntamento stabilito. Il gruppo controllo è stato sottoposto invece ad un normale approccio odontoiatrico. Risultati: Il metodo educativo TEACH ci ha permesso di poter far affrontare con serenità ad 8 dei nostri pazienti appartenenti al gruppo test la visita odontoiatrica e l’esecuzione di metodiche atte alla prevenzione. In particolare il livello di collaborazione è stato ottimo per 3 degli 8 pazienti e buono per gli altri 5. Per 3 pazienti la collaborazione si è dimostrata, nonostante il tipo di approccio, scarsa e comunque insufficiente per le cure odontoiatriche ambulatoriali. Nel gruppo di controllo la collaborazione si è dimostrata scarsa ed insufficiente per le cure ambulatoriali in 7 casi su 11.Conclusioni: L’approccio alle cure odontoiatriche con il metodo TEACH ha permesso l’accesso all’ambulatorio odontoiatrico per la visita e le cure preventive al 72% di un gruppo di pazienti con DPS, che erano risultati in precedenza non collaboranti. Invece, nel gruppo di controllo i pazienti selezionati per le cure ambulatoriali sono risultati essere solo il 36%. A nostro parere, il metodo di approccio proposto risulta efficace nel ridurre l’”ansia da esposizione e da rottura degli schemi” in pazienti con DP

La riabilitazione protesica di un caso di Displasia Ectodermica Anidrotica (HED)

The absolute intensities of cosmic ray muons in the vertical direction and at several large zenith angles in the geomagnetic west have been measured for momenta in the range 3-100 GeV/c by use of a solid-iron magnetic momentum spectrometer-telescope. Details of the experimental apparatus are presented, and the methods of measurement and analysis of the basic data from photographic re-projection of film records of events in wide-gap spark chambers are discussed. The intensities of the present experiment are systematically ~15-40% higher than previously reported intensities at comparable momenta and zenith angles. Reasons for believing that the present work is of improved accuracy in comparison with previous experimental work and actually gives absolute intensities are presented. The impact of these results on several areas of high-energy nuclear and cosmic ray physics is discussed