Role of Grb10 in mTORC1-dependent regulation of insulin signaling and action in human skeletal muscle cells.

Growth factor receptor-bound protein 10 (Grb10) is an adaptor protein that binds to the insulin receptor, upon which insulin signaling and action are thought to be inhibited. Grb10 is also a substrate for the mechanistic target of rapamycin complex 1 (mTORC1) that mediates its feedback inhibition on phosphatidylinositide 3-kinase (PI3K)/Akt signaling. To characterize the function of Grb10 and its regulation by mTORC1 in human muscle, primary skeletal muscle cells were isolated from healthy lean young men and then induced to differentiate into myotubes. Knockdown of Grb10 enhanced insulin-induced PI3K/Akt signaling and glucose uptake in myotubes, reinforcing the notion underlying its function as a negative regulator of insulin action in human muscle. The increased insulin responsiveness in Grb10-silenced myotubes was associated with a higher abundance of the insulin receptor. Furthermore, insulin and amino acids independently and additively stimulated phosphorylation of Grb10 at Ser476. However, acute inhibition of mTORC1 with rapamycin blocked Grb10 Ser476 phosphorylation and repressed a negative-feedback loop on PI3K/Akt signaling that increased myotube responsiveness to insulin. Chronic rapamycin treatment reduced Grb10 protein abundance in conjunction with increased insulin receptor protein levels. Based on these findings, we propose that mTORC1 controls PI3K/Akt signaling through modulation of insulin receptor abundance by Grb10. These findings have potential implications for obesity-linked insulin resistance, as well as clinical use of mTORC1 inhibitors.

Changes detected in the genome sequences of Escherichia coli, Listeria monocytogenes, Vibrio parahaemolyticus, and Salmonella enterica after serial subculturing.

Whole genome sequencing (WGS) is rapidly replacing other molecular techniques for identifying and subtyping bacterial isolates. The resolution or discrimination offered by WGS is significantly higher than that offered by other molecular techniques, and WGS readily allows infrequent differences that occur between 2 closely related strains to be found. In this investigation, WGS was used to identify the changes that occurred in the genomes of 13 strains of bacterial foodborne pathogens after 100 serial subcultures. Pure cultures of Shiga-toxin-producing Escherichia coli, Salmonella enterica, Listeria monocytogenes, and Vibrio parahaemolyticus were subcultured daily for 100 successive days. The 1st and 100th subcultures were whole-genome sequenced using short-read sequencing. Single nucleotide polymorphisms (SNPs) were identified between the 1st and final culture using 2 different approaches, and multilocus sequence typing of the whole genome was also performed to detect any changes at the allelic level. The number of observed genomic changes varied by strain, species, and the SNP caller used. This study provides insight into the genomic variation that can be detected using next-generation sequencing and analysis methods after repeated subculturing of 4 important bacterial pathogens.

Partial substitutions of animal with plant protein foods in Canadian diets have synergies and trade-offs among nutrition, health and climate outcomes.

Dietary guidelines emphasize the consumption of plant protein foods, but the implications of replacing animal with plant sources on a combination of diet sustainability dimensions are unknown. Using a combination of data from a national nutrition survey, greenhouse gas emissions from dataFIELD and relative risks from the Global Burden of Disease Study 2017, we assess the impact of partially substituting red and processed meat or dairy with plant protein foods in Canadian self-selected diets on nutrition, health and climate outcomes. The substitutions induced minor changes to the percentage of the population below requirements for nutrients of concern, but increased calcium inadequacy by up to 14% when dairy was replaced. Replacing red and processed meat or dairy increased life expectancy by up to 8.7 months or 7.6 months, respectively. Diet-related greenhouse gas emissions decreased by up to 25% for red and processed meat and by up to 5% for dairy replacements. Co-benefits of partially substituting red and processed meat with plant protein foods among nutrition, health and climate outcomes are relevant for reshaping consumer food choices in addressing human and planetary health.

Protein consumption in Canadian habitual diets: usual intake, inadequacy, and the contribution of animal- and plant-based foods to nutrient intakes.

The 2019 Canada's Food Guide (CFG) emphasizes consumption of plant protein with implications for protein adequacy and nutrient intakes, yet a baseline with which to compare future dietary trends that may result from its adoption is not available. The objectives were to assess usual protein intake, inadequacy, and the contribution of animal- and plant-based foods to intake of protein, nutrients, and energy in Canada. Twenty-four-hour dietary recalls from the 2015 Canadian Community Health Survey - Nutrition were used to assess dietary intake among adults (n = 13 616). The National Cancer Institute method was used to estimate usual protein intake and inadequacy. Population ratios were used to determine the contribution of animal- and plant-based foods to intake of protein, nutrients, and energy. Usual protein intake averaged 79.47 ± 0.70 g/d; inadequacy was highest for females ≥71 y (9.76 ± 2.04%). Top protein contributors were red and processed meat (21.6 ± 0.55%), poultry and eggs (20.1 ± 0.81%), cereals, grains, and breads (19.5 ± 0.31%), and dairy (16.7 ± 0.38%). Dairy contributed most to calcium (53.4 ± 0.61%), vitamin D (38.7 ± 1.01%), but also saturated fat (40.6 ± 0.69%), whereas cereals, grains, and breads contributed most to iron (46.5 ± 0.57%) and vegetables and fruit to potassium (32 ± 0.45%). Given that animal sources contributed overwhelmingly to protein intake in 2015, dietary shifts towards plant protein needed to meet the 2019 CFG recommendations may pose a challenge, particularly for populations most at risk of inadequacy. Novelty: Older adults and females are most at risk of not meeting protein recommendations. Animal sources contribute two-thirds of the protein consumed by Canadian adults.

Consumption of Milk and Alternatives and Their Contribution to Nutrient Intakes among Canadian Adults: Evidence from the 2015 Canadian Community Health Survey-Nutrition.

As a staple food and dense source of nutrients, milk and alternatives play an important role in nutrient adequacy. The aims of this study were to quantify the consumption of milk and alternatives within Canadian self-selected diets and determine their contribution to intakes of nutrients and energy. First, 24-h dietary recalls from the 2015 Canadian Community Health Survey-Nutrition were used to assess 1-d food and nutrient intakes among Canadian adults ≥19 y (n = 13,616). Foods were classified as milk and alternatives according to the 2007 Canada's Food Guide. Descriptive statistics were used to calculate daily servings of milk and alternatives by different age groups and demographic characteristics. Population ratios were used to discern their contribution to total intakes of nutrients and energy. Mean daily servings (±SE) were highest for milk (0.60 ± 0.02) and cheese (0.42 ± 0.01), intermediate for frozen dairy (0.16 ± 0.01) and yoghurt (0.14 ± 0.01), and lowest for soy and other dairy (<0.03). Intakes were lowest among Canadians 51+ y (1.3 ± 0.03), females (1.25 ± 0.03), non-Caucasians (1.06 ± 0.05), those with less than a secondary education (1.19 ± 0.05), and British Columbians (1.17 ± 0.05). Milk and alternatives contributed >20% to total intakes of calcium (52.62 ± 0.46%), vitamin D (38.53 ± 0.78%), saturated fat (28.84 ± 0.51%), vitamin B12 (27.73 ± 0.57%), vitamin A (26.16 ± 0.58%), phosphorus (24.76 ± 0.35%), and riboflavin (24.43 ± 0.37%), of which milk was the top source. Milk and alternatives contribute substantially to nutrient intakes and thus warrant further attention in terms of mitigating nutrient inadequacy among the Canadian population.