Cytotoxic Aggregation and Amyloid Formation by the Myostatin Precursor Protein

Myostatin, a negative regulator of muscle growth, has been implicated in sporadic inclusion body myositis (sIBM). sIBM is the most common age-related muscle-wastage disease with a pathogenesis similar to that of amyloid disorders such as Alzheimer's and Parkinson's diseases. Myostatin precursor protein (MstnPP) has been shown to associate with large molecular weight filamentous inclusions containing the Alzheimer's amyloid beta peptide in sIBM tissue, and MstnPP is upregulated following ER stress. The mechanism for how MstnPP contributes to disease pathogenesis is unknown. Here, we show for the first time that MstnPP is capable of forming amyloid fibrils in vitro. When MstnPP-containing Escherichia coli inclusion bodies are refolded and purified, a proportion of MstnPP spontaneously misfolds into amyloid-like aggregates as characterised by electron microscopy and binding of the amyloid-specific dye thioflavin T. When subjected to a slightly acidic pH and elevated temperature, the aggregates form straight and unbranched amyloid fibrils 15 nm in diameter and also exhibit higher order amyloid structures. Circular dichroism spectroscopy reveals that the amyloid fibrils are dominated by β-sheet and that their formation occurs via a conformational change that occurs at a physiologically relevant temperature. Importantly, MstnPP aggregates and protofibrils have a negative effect on the viability of myoblasts. These novel results show that the myostatin precursor protein is capable of forming amyloid structures in vitro with implications for a role in sIBM pathogenesis

Priority nutrients to address malnutrition and diet-related diseases in Australia and New Zealand

Background: The double burden of malnutrition and diet-related disease has been attributed to diets high in ultra-processed and discretionary foods, with increased sugars, saturated fats, and sodium, and insufficient dietary fibre. There is a limited understanding of the role of other macronutrients and micronutrients. Objective: Determine the highest priority nutrients to address both malnutrition and diet-related disease in Australia and New Zealand, for each demographic group and the total population. Methods: A novel four-step methodological approach was undertaken to identify: 1. Demographic (age-sex) groups; 2. Health priorities; 3. Potential nutrients based on inadequacy, increased requirements, and health priority association; and 4. Priority nutrients. Nutrient intake data was obtained from the most recent Australian and New Zealand nutrition surveys. Health priorities were based on national statistical data and expert consultation. High-level scientific literature (systematic reviews) was scoped for associations with health priorities and the suitability of recommended intakes. A quantitative scoring matrix was developed and used to determine the highest priority nutrients, with scoring over three domains: extent of inadequacy; consensus for increased requirements; and degree of association with health priorities. Results: Nutritional inadequacies were common, with 22 of 31 essential nutrients consumed below recommended levels. Nine priority nutrients were identified across the demographic groups, with each demographic group characterised by a specific subset of these. Six nutrients were highest priority within the total population: vitamin D, calcium, omega-3 fatty acids, magnesium, folate, dietary fibre. Conclusion: The extent of nutritional inadequacies in Australia and New Zealand is high, both within each demographic group and the entire population, relative to both recommended intakes and key health outcomes. The methodology can be applied to other countries and globally. Findings make a significant contribution to understanding the nutrients to prioritise in future-proofing the health of the Australian and New Zealand populations. Guidelines and policies can target priority nutrients to address the malnutrition and diet-related disease double burden.</p

Diet and Economic Modelling to Improve the Quality and Affordability of the Australian Diet for Low and Medium Socioeconomic Households

Food costs are a barrier to healthier diet selections, particularly for low socioeconomic households who regularly choose processed foods containing refined grains, added sugars, and added fats. In this study, the objectives were to: (i) identify the nutrient density-to-cost ratio of Australian foods; (ii) model the impact of substituting foods with lower nutrient density-to-cost ratio with those with the highest nutrient density-to-cost ratio for diet quality and affordability in low and medium socioeconomic households; and (iii) evaluate food processing levels. Foods were categorized, coded for processing level, analysed for nutrient density and cost, and ranked by nutrient density-to-cost ratio. The top quartile of nutrient dense, low-cost foods included 54% unprocessed (vegetables and reduced fat dairy), 33% ultra-processed (fortified wholegrain bread and breakfast cereals <20 g sugars/100 g), and 13% processed (fruit juice and canned legumes). Using substitution modelling, diet quality improved by 52% for adults and 71% for children across all households, while diet affordability improved by 25% and 27% for low and medium socioeconomic households, respectively. The results indicate that the quality and affordability of the Australian diet can be improved when nutritious, low-cost foods are selected. Processing levels in the healthier modelled diets suggest that some ultra-processed foods may provide a beneficial source of nutrition when consumed within national food group recommendations

Nutrient dense, low-cost foods can improve the affordability and quality of the new zealand diet—a substitution modeling study

The high prevalence of non-communicable disease in New Zealand (NZ) is driven in part by unhealthy diet selections, with food costs contributing to an increased risk for vulnerable population groups. This study aimed to: (i) identify the nutrient density-to-cost ratio of NZ foods; (ii) model the impact of substituting foods with a lower nutrient density-to-cost ratio with those with a higher nutrient density-to-cost ratio on diet quality and affordability in representative NZ population samples for low and medium socioeconomic status (SES) households by ethnicity; and (iii) evaluate food processing level. Foods were categorized, coded for processing level and discretionary status, analyzed for nutrient density and cost, and ranked by nutrient density-to-cost ratio. The top quartile of nutrient dense, low-cost foods were 56% unprocessed (vegetables, fruit, porridge, pasta, rice, nuts/seeds), 31% ultra-processed (vegetable dishes, fortified bread, breakfast cereals unfortified <15 g sugars/100 g and fortified 15–30 g sugars/100 g), 6% processed (fruit juice), and 6% culinary processed (oils). Using substitution modeling, diet quality improved by 59% and 71% for adults and children, respectively, and affordability increased by 20–24%, depending on ethnicity and SES. The NZ diet can be made healthier and more affordable when nutritious, low-cost foods are selected. Processing levels in the healthier, modeled diet suggest that some non-discretionary ultra-processed foods may provide a valuable source of low-cost nutrition for food insecure populations

The C313Y Piedmontese mutation decreases myostatin covalent dimerisation and stability

<p>Abstract</p> <p>Background</p> <p>Myostatin is a key negative regulator of muscle growth and development, whose activity has important implications for the treatment of muscle wastage disorders. Piedmontese cattle display a double-muscled phenotype associated with the expression of C313Y mutant myostatin. <it>In vivo</it>, C313Y myostatin is proteolytically processed, exported and circulated extracellularly but fails to correctly regulate muscle growth. The C313Y mutation removes the C313-containing disulphide bond, an integral part of the characteristic TGF-β cystine-knot structural motif.</p> <p>Results</p> <p>Here we present <it>in vitro </it>analysis of the structure and stability of the C313Y myostatin protein that reveals significantly decreased covalent dimerisation for C313Y myostatin accompanied by a loss of structural stability compared to wild type. The C313Y myostatin growth factor, processed from full length precursor protein, fails to inhibit C2C12 myoblast proliferation in contrast to wild type myostatin. Although structural modeling shows the substitution of tyrosine causes structural perturbation, biochemical analysis of additional disulphide mutants, C313A and C374A, indicates that an intact cystine-knot motif is a major determinant in myostatin growth factor stability and covalent dimerisation.</p> <p>Conclusions</p> <p>This research shows that the cystine-knot structure is important for myostatin dimerisation and stability, and that disruption of this structural motif perturbs myostatin signaling.</p

The human myostatin precursor protein : structure, function and amyloid formation : implications for the muscle wastage disease sporadic inclusion body myositis : a dissertation presented in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Biochemistry at Massey University, Palmerston North, New Zealand

Myostatin is a major player in the regulation of mammalian muscle growth and development, maintaining the balance between proliferation and differentiation prenatally and the quiescence of satellite cells in adults. An absence or overexpression of myostatin results in double-muscling and cachexia respectively, placing myostatin as a promising target in the treatment of muscle wastage diseases. As a transforming growth factor-β superfamily member, myostatin is produced as a precursor protein, consisting of a propeptide region N-terminal to the growth factor domain. Cleavage of the precursor between the domains forms the myostatin latent complex, an inhibitory structure which is exported from the cell where a second cleavage event releases the active myostatin growth factor. The precursor protein, propeptide, and latent complex play important roles in the regulation of myostatin. However, their structure and function are poorly understood, and a possible role for the myostatin precursor protein in the muscle wastage disease sporadic inclusion body myositis, suggests that pre-growth factor forms of myostatin may be additional important therapeutic targets. This thesis presents an investigation into the structure and function of the myostatin precursor protein, the latent complex, and the propeptide region within these, with comparisons to a mutant form of myostatin responsible for the naturally-occurring double-muscled phenotype of the Piedmontese cattle breed. Results suggest that the diverse functions of the propeptide region are facilitated by regions of intrinsic disorder within a primarily structured domain, and that conformational alterations accompany the precursor to latent complex transition, resulting in a tighter inhibitory structure. Comparative analyses between the wild-type and mutant proteins suggest that the Piedmontese phenotype is due to a reduced capacity for covalent dimerisation and significant structural alterations within the type I receptor-binding domain. Investigation into misfolded myostatin precursor protein found that the precursor is able to form cytotoxic amyloid aggregates and mature fibrils under partially denaturing conditions, suggesting a possible mechanism for the role of the myostatin precursor in sporadic inclusion body myositis. Together, these novel results contribute important information towards an understanding of myostatin structure, function and regulation in both normal and disease scenarios

Should We ‘Eat a Rainbow’? An Umbrella Review of the Health Effects of Colorful Bioactive Pigments in Fruits and Vegetables

Health promotion campaigns have advocated for individuals to ‘eat a rainbow’ of fruits and vegetables (FV). However, the literature has only focused on individual color pigments or individual health outcomes. This umbrella review synthesized the evidence on the health effects of a variety of color-associated bioactive pigments found in FV (carotenoids, flavonoids, betalains and chlorophylls), compared to placebo or low intakes. A systematic search of PubMed, EMBASE, CINAHL and CENTRAL was conducted on 20 October 2021, without date limits. Meta-analyzed outcomes were evaluated for certainty via the GRADE system. Risk of bias was assessed using the Centre for Evidence-Based Medicine critical appraisal tools. A total of 86 studies were included, 449 meta-analyzed health outcomes, and data from over 37 million participants were identified. A total of 42% of health outcomes were improved by color-associated pigments (91% GRADE rating very low to low). Unique health effects were identified: n = 6 red, n = 10 orange, n = 3 yellow, n = 6 pale yellow, n = 3 white, n = 8 purple/blue and n = 1 green. Health outcomes associated with multiple color pigments were body weight, lipid profile, inflammation, cardiovascular disease, mortality, type 2 diabetes and cancer. Findings show that color-associated FV variety may confer additional benefits to population health beyond total FV intake

Data_Sheet_1_Mushrooms: a food-based solution to vitamin D deficiency to include in dietary guidelines.pdf

Vitamin D deficiency and insufficiency is a public health issue, with low dietary vitamin D intakes a contributing factor. Rates of vitamin D deficiency are 31% in Australia, and up to 72% in some regions globally. While supplementation is often prescribed as an alternative to additional sun exposure, complementary approaches including food-based solutions are needed. Yet, food-centric dietary guidelines are not always adequate for meeting vitamin D needs. Edible mushrooms such as Agaricus bisporus can produce over 100% of vitamin D recommendations (10 μg/day, Institute of Medicine) per 75 g serve (18 μg) on exposure to UV-light, with the vitamin D2 produced showing good stability during cooking and processing. However, mushrooms are overlooked as a vitamin D source in dietary guidelines. Our dietary modelling shows that four serves/week of UV-exposed button mushrooms can support most Australian adults in meeting vitamin D recommendations, and UV-exposed mushrooms have been found to increase vitamin D status in deficient individuals. While recent evidence suggests some differences between vitamin D2 and vitamin D3 in physiological activities, vitamin D2 from mushrooms can be part of a larger solution to increasing dietary vitamin D intakes, as well as an important focus for public health policy. Mushrooms exposed to UV represent an important tool in the strategic toolkit for addressing vitamin D deficiency in Australia and globally. Health authorities lead the recognition and promotion of mushrooms as a natural, vegan, safe, and sustainable vitamin D food source.</p