Protein hydrolysates from boarfish (Capros aper) and Atlantic salmon (Salmo salar) skin gelatin improve metabolic control in genetically obese diabetic (ob/ob) mice

There is increasing interest in dietary protein for management of Type 2 diabetes mellitus (T2DM) and obesity. The effects of twice-daily oral administration of a salmon skin gelatin hydrolysate (SSGH, 50 mg/kg), boarfish protein hydrolysate (BPH, (50 mg/kg), metformin (200 mg/kg), or saline control, were investigated in ob/ob mice. Non-fasting blood glucose was significantly reduced with SSGH (p < 0.01), BPH (p < 0.001) and metformin (p < 0.001), which were reflected in reductions in glycated haemoglobin (HbA1c) (p < 0.001, p < 0.01 and p < 0.01, respectively). Responses to oral and intraperitoneal glucose tolerance were improved (p < 0.05–0.01), as well as circulating plasma lipid profiles (p < 0.05–0.001). Chronic BPH treatment increased circulating plasma insulin (p < 0.01), whereas SSGH improved insulin sensitivity (p < 0.05), versus respective controls. All treatments significantly reduced energy intake (p < 0.05–0.001) versus (ob/ob) controls, without affecting overall bodyweight. These findings suggest that fish hydrolysates mediate potent anti-diabetic actions similar to metformin and might be suitable for the management and prevention of T2DM

A narrative review of the anti-hyperglycemic and satiating effects of fish protein hydrolysates and their bioactive peptides

Prevalence of type 2 diabetes and overweight/obesity are increasing globally. Food supplementation as a preventative option has become an attractive option in comparison to increased pharmacotherapy dependency. Hydrolysates of fish processing waste and by-products have become particularly interesting in a climate of increased food wastage awareness and are rapidly gaining traction in food research. This review summarizes the available research so far on the potential effect of these hydrolysates on diabetes and appetite suppression. Scopus and Web of Science are searched using eight keywords (fish, hydrolysate, peptides, satiating, insulinotropic, incretin, anti-obesity, DPP-4 [dipeptidylpeptidase-4/IV]) returning a total of 2549 results. Following exclusion criteria (repeated appearances, non-fish marine sources [e.g., macroalgae], and irrelevant bioactivities [e.g., immunomodulatory, anti-thrombotic]), 44 relevant publications are included in this review. Stimulation of hormone secretion, regulation of glucose uptake, anorexigenic potential, identified mechanisms of action, and research conducted on the most potent bioactive peptides identified within these hydrolysates are all specifically addressed. Results of this review conclude that despite wide methodological variation between studies, there is significant potential for the application of fish protein hydrolysates in the management of bodyweight and hyperglycemia

Physicochemical, nutritional and in vitro antidiabetic characterisation of blue whiting (micromesistius poutassou) protein hydrolysates

Protein hydrolysates from low-value underutilised fish species are potential sources of high quality dietary protein and health enhancing peptides. Six blue whiting soluble protein hydrolysates (BW-SPH-A_F), generated at industrial scale using different hydrolysis conditions, were assessed in terms of their protein equivalent content, amino acid profile and score and physicochemical properties in addition to their ability to inhibit dipeptidyl peptidase IV (DPP-IV) and stimulate the secretion of insulin from BRIN-BD11 cells. Furthermore, the effect of simulated gastrointestinal digestion (SGID) on the stability of the BW-SPHs and their associated in vitro antidiabetic activity was investigated. The BW-SPHs contained between 70–74% (w/w) protein and all essential and non essential amino acids. All BW-SPHs mediated DPP-IV inhibitory (IC50: 2.12–2.90 mg protein/mL) and insulin secretory activity (2.5 mg/mL; 4.7 to 6.4-fold increase compared to the basal control (5.6 mM glucose alone)). All BW-SPHs were further hydrolysed during SGID. While the in vitro DPP-IV inhibitory and insulin secretory activity mediated by some BW-SPHs was reduced following SGID, the activity remained high. In general, the insulin secretory activity of the BW-SPHs were 4.5–5.4-fold higher than the basal control following SGID. The BW-SPHs generated herein provide potential for anti-diabetic related functional ingredients, whilst also enhancing environmental and commercial sustainability

Twice daily oral administration of Palmaria palmata protein hydrolysate reduces food intake in streptozotocin induced diabetic mice, improving glycaemic control and lipid profiles

This study investigated the antihyperglycaemic effectiveness of an oral Palmaria palmata protein hydrolysate (PPPH), versus metformin, upon metabolic control in streptozotocin (STZ)-induced diabetic mice. Mice were administered PPPH (50 mg/kg bodyweight) or metformin (200 mg/kg bodyweight) by oral gavage twice-daily for 18 days. Blood glucose and plasma insulin were measured every third day. PPPH caused a significant reduction in blood glucose (p < 0.001) and a significant increase in plasma insulin (p < 0.001) versus STZtreated saline controls. PPPH treatment reduced energy intake (p < 0.05), bodyweight (p < 0.01) and total plasma glucagon-like peptide-1 (p < 0.01) after 18 days. Terminal oral glucose tolerance (Day 18, p < 0.05), fasting blood glucose (p < 0.001), HbA1C (p < 0.01), plasma cholesterol (p < 0.01) and plasma triglycerides (p < 0.05) were significantly improved versus STZ-treated saline controls. All groups showed significant increases in pancreatic islet area, β-cell area, and β:α cell ratio. PPPH demonstrated potent antidiabetic potential in vivo through reduced food intake and improved beta-cell function

Macroalgal protein hydrolysates from palmaria palmata influence the ‘incretin effect’ in vitro via DPP‑4 inhibition and upregulation of insulin, GLP‑1 and GIP secretion

Purpose This study investigated metabolic benefits of protein hydrolysates from the macroalgae Palmaria palmata, previously shown to inhibit dipeptidylpeptidase-4 (DPP-4) activity in vitro. Methods Previously, Alcalase/Flavourzyme-produced P. palmata protein hydrolysate (PPPH) improved glycaemia and insulin production in streptozotocin-induced diabetic mice. Here the PPPH, was compared to alternative Alcalase, bromelain and Promod-derived hydrolysates and an unhydrolysed control. All PPPH’s underwent simulated gastrointestinal digestion (SGID) to establish oral bioavailability. PPPH’s and their SGID counterparts were tested in pancreatic, clonal BRIN-BD11 cells to assess their insulinotropic efect and associated intracellular mechanisms. PPPH actions on the incretin effect were assessed via measurement of DPP-4 activity, coupled with GLP-1 and GIP release from GLUTag and STC-1 cells, respec tively. Acute in vivo effects of Alcalase/Flavourzyme PPPH administration on glucose tolerance and satiety were assessed in overnight-fasted mice. Results PPPH’s (0.02–2.5 mg/ml) elicited varying insulinotropic efects (p<0.05–0.001). SGID of the unhydrolysed protein control, bromelain and Promod PPPH’s retained, or improved, bioactivity regarding insulin secretion, DPP-4 inhibition and GIP release. Insulinotropic effects were retained for all SGID-hydrolysates at higher PPPH concentrations. DPP-4 inhibitory effects were confrmed for all PPPH’s and SGID counterparts (p<0.05–0.001). PPPH’s were shown to directly influence the incretin effect via upregulated GLP-1 and GIP (p<0.01–0.001) secretion in vitro, largely retained after SGID. Alcalase/Flavourzyme PPPH produced the greatest elevation in cAMP (p<0.001, 1.7-fold), which was fully retained post-SGID. This hydrolysate elicited elevations in intracellular calcium (p<0.01) and membrane potential (p<0.001). In acute in vivo settings, Alcalase/Flavourzyme PPPH improved glucose tolerance (p<0.01–0.001) and satiety (p<0.05–0.001). Conclusion Bioavailable PPPH peptides may be useful for the management of T2DM and obesity

Identification and characterisation of peptides from a boarfish (Capros aper) protein hydrolysate displaying in vitro dipeptidyl peptidase-IV (DPP-IV) inhibitory and insulinotropic activity

Twenty-two novel dipeptidyl peptidase-IV (DPP-IV) inhibitory peptides (with IC50 values <200 µM) and fifteen novel insulinotropic peptides were identified in a boarfish protein hydrolysate generated at semi-pilot scale using Alcalase 2.4L and Flavourzyme 500L. This was achieved by bioassay-driven semi preparative reverse phase-high performance liquid chromatography fractionation, liquid chromatography-mass spectrometry and confirmatory studies with synthetic peptides. The most potent DPP-IV inhibitory peptide (IPVDM) had a DPP-IV half maximal inhibitory concentration (IC50) values of 21.72 ± 1.08 µM in a conventional in vitro and 44.26 ± 0.65 µM in an in situ cell-based (Caco 2) DPP-IV inhibition assay. Furthermore, this peptide stimulated potent insulin secretory activity (1.6 fold increase compared to control) from pancreatic BRIN-BD11 cells grown in culture. The tripeptide IPV exhibited potent DPP-IV inhibitory activity (IC50: 5.61 ± 0.20 µM) comparable to that reported for the known DPP-IV inhibitor IPI (IC50: 5.61 µM). Boarfish proteins contain peptide sequences with potential to play a role in glycaemic management in vivo

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

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