Citrus polyphenols and risk of type 2 diabetes: Evidence from mechanistic studies.

Citrus fruits are a rich source of (poly)phenols, a group of dietary bioactive compounds that protect against developing type 2 diabetes. Our review critically evaluates how experimental in vitro and animal models have elucidated some of the underlying mechanisms on how citrus (poly)phenols affect the markers of type 2 diabetes. According to animal studies, the beneficial effects derived from consuming citrus compounds appear to be related to long-term effects, rather than acute. There are some notable effects from citrus (poly)phenol metabolites on post-absorptive processes, such as modulation of hepatic glucose metabolism and insulin sensitivity in target tissues, but with a more modest effect on digestion and sugar absorption within the gut. Experimental studies on cells and other systems in vitro have indicated some of the possible mechanisms involved, but ∼70% of the studies utilized unrealistically high concentrations and forms of the compounds, compromising physiological relevance. Future studies should discuss the relevance of concentration used in in vitro experiments, relative to the proposed site of action, and also examine the role of catabolites produced by the gut microbiota. Finally, it is important to examine the relationship between the gut microbiota and bioavailability on the action of citrus (poly)phenols.

Review of factors affecting citrus polyphenol bioavailability and their importance in designing in vitro, animal, and intervention studies.

Evidence from in vitro, animal, and human studies links citrus fruit consumption with several health-promoting effects. However, many in vitro studies disregard bioavailability data, a key factor determining responses in humans. Citrus (poly)phenol metabolism and bioavailability follow specific pathways that vary widely among individuals and are affected by several intrinsic (age, sex, gut microbiota, metabolic state, genetic polymorphisms) and extrinsic (food matrix, co-consumed food, (poly)phenol solubility, dose, food processing, lifestyle) factors. The gut microbiota is crucial to both absorption of citrus (poly)phenols and the production of catabolites, and absorption of both takes place mostly in the colon. Citrus (poly)phenol absorption can reach up to 100% in some individuals when the sum of the gut microbiota products are taken into account. This review emphasizes the importance of understanding citrus (poly)phenol absorption, metabolism, and bioavailability using evidence primarily derived from human studies in designing in vitro, animal, and further human clinical studies.

Measuring key human carbohydrate digestive enzyme activities using high-performance anion-exchange chromatography with pulsed amperometric detection.

Carbohydrate digestion in the mammalian gastrointestinal tract is catalyzed by α-amylases and α-glucosidases to produce monosaccharides for absorption. Inhibition of these enzymes is the major activity of the drugs acarbose and miglitol, which are used to manage diabetes. Furthermore, delaying carbohydrate digestion via inhibition of α-amylases and α-glucosidases is an effective strategy to blunt blood glucose spikes, a major risk factor for developing metabolic diseases. Here, we present an in vitro protocol developed to accurately and specifically assess the activity of α-amylases and α-glucosidases, including sucrase, maltase and isomaltase. The assay is especially suitable for measuring inhibition by compounds, drugs and extracts, with minimal interference from impurities or endogenous components, because the substrates and digestive products in the enzyme activity assays are quantified directly by high-performance anion-exchange chromatography with pulsed amperometric detection (HPAE-PAD). Multiple enzyme sources can be used, but here we present the protocol using commercially available human α-amylase to assess starch hydrolysis with maltoheptaose as the substrate, and with brush border sucrase-isomaltase (with maltase, sucrase and isomaltase activities) derived from differentiated human intestinal Caco-2(/TC7) cells to assess hydrolysis of disaccharides. The wet-lab assay takes ~2-5 h depending on the number of samples, and the HPAE-PAD analysis takes 35 min per sample. A full dataset therefore takes 1-3 d and allows detection of subtle changes in enzyme activity with high sensitivity and reliability.

Chemical composition and health benefits of coconut oil: an overview.

Coconut oil is an integral part of Sri Lankan and many South Asian diets. Initially, coconut oil was classified along with saturated fatty acid food items and criticized for its negative impact on health. However, research studies have shown that coconut oil is a rich source of medium-chain fatty acids. Thus, this has opened new prospects for its use in many fields. Beyond its usage in cooking, coconut oil has attracted attention due to its hypocholesterolemic, anticancer, antihepatosteatotic, antidiabetic, antioxidant, anti-inflammatory, antimicrobial and skin moisturizing properties. Despite all the health benefits, consumption of coconut oil is still underrated due to a lack of supportive scientific evidence. Even though studies done in Asian countries claim a favorable impact on cardiac health and serum lipid profile, the limitations in the number of studies conducted among Western countries impede the endorsement of the real value of coconut oil. Hence, long-term extensive studies with proper methodologies are suggested to clear all the controversies and misconceptions of coconut oil consumption. This review discusses the composition and functional properties of coconut oils extracted using various processing methods. © 2020 Society of Chemical Industry.

Maltoheptaoside hydrolysis with chromatographic detection and starch hydrolysis with reducing sugar analysis: Comparison of assays allows assessment of the roles of direct α-amylase inhibition and starch complexation.

The aim was to determine inhibition of human α-amylase activity by (poly)phenols using maltoheptaoside as substrate with direct chromatographic product quantification, compared to hydrolysis of amylose and amylopectin estimated using 3,5-dinitrosalicylic acid. Acarbose exhibited similar IC50 values (50% inhibition) with maltoheptaoside, amylopectin or amylose as substrates (2.37 ± 0.11, 3.71 ± 0.12 and 2.08 ± 0.01 µM respectively). Epigallocatechin gallate, quercetagetin and punicalagin were weaker inhibitors of hydrolysis of maltoheptaoside (<50% inhibition) than amylose (IC50: epigallocatechin gallate = 20.41 ± 0.25 µM, quercetagetin = 30.15 ± 2.05 µM) or amylopectin. Interference using 3,5-dinitrosalicylic acid was in the order punicalagin > epigallocatechin gallate > quercetagetin, with minimal interference using maltoheptaoside as substrate. The main inhibition mechanism of epigallocatechin gallate and punicalagin was through complexation with starch, especially amylose, whereas only quercetagetin additionally binds to the α-amylase active site. Interference is minimised using maltoheptaoside as substrate with product detection by chromatography, potentially allowing assessment of direct enzyme inhibition by almost any compound.

Critical review on conventional spectroscopic α-amylase activity detection methods: merits, demerits, and future prospects.

α-Amylase is an endoenzyme that catalyses the hydrolysis of internal α-l,4 glycosidic linkages in polysaccharides to produce maltose, maltotriose, and α-limit dextrins. It is widely used in the laboratorial and industrial workflow for several applications. There are several methods utilizing different techniques and substrates to assess α-amylase activity, among which the spectroscopic methods have found widespread applicability due to their ease of use and cost-effectiveness. Depending upon the reaction principle, these assays are classified into four groups: reducing sugar, enzymatic, chromogenic, and amyloclastic methods. Despite the presence of numerous methods, there is no general reliable method to assess α-amylase activity. Each method is shown to have its own merits and demerits. Many improvements have been made to make the available methods more accurate, reliable, and easy. This communication briefly discusses the basic reaction mechanisms and critically reviews the advantages and shortcomings associated with each method. Further recommendations are made for future development. © 2020 Society of Chemical Industry.

Applicability and reliability of the glucose oxidase method in assessing α-amylase activity.

Glucose oxidase (GOD) is an enzyme widely used in glucose monitoring systems owing to its high specificity towards glucose. However, in our previous work maltose was found to show significant interaction with GOD and based on this observation, a novel microplate-based method was developed to assess α-amylase inhibitory activity (GOD method). Concerns regarding the interaction of GOD with maltose has limited the widespread use of the GOD method in assessing α-amylase activity. The present paper provides answers to concerns regarding the interaction of GOD with maltose using HPLC studies and application of the GOD method in assessing α-amylase activity. According to the results, the newly developed GOD method can be considered as a well-suited method for the determination of α-amylase activity and as an easy method to do kinetic studies compared to other available methods.

Boiled, sprouted, and raw cowpea-incorporated diets modulate high-fat diet-induced hypercholesterolemia in rats.

This study was carried out to investigate the effect of processed (boiled and sprouted) cowpea-incorporated experimental diets on serum cholesterol and serum antioxidant capacity in high-fat diet (HFD)-fed Wistar rats. Seven weeks old male Wistar rats were fed 20% fat as a control (CD), for comparison with 20% fat-enriched diets containing 20% whole raw cowpea diets (Bombay Raw Diet; BRD and MI35 Raw Diet; MRD), boiled cowpea diets (Bombay Boiled Diet; BBD and MI35 Boiled Diet; MBD) and sprouted cowpea diet (Bombay Sprouted Diet; BSD) for 6 weeks. The increase in serum total cholesterol as a result of high-fat diet was significantly countered by boiled and raw cowpea-incorporated diet-fed rats. Increased serum non-HDL-C level caused by HFD was significantly (p < 0.05) countered by raw, boiled, and sprouted cowpeas, while HDL-C was increased by raw MI and boiled Bombay incorporated diets. Boiling has improved the hypocholesterolemic ability of Bombay cowpea and BBD has significantly (p < 0.05) modulated serum HDL-C level and liver weight in rats. These findings were supported significantly high soluble fiber content in processed cowpea powder than that in raw cowpea powder. The decrease in serum antioxidant activity as a result of HFD was significantly countered by BRD. Processing has reduced the antioxidant activity in cowpeas and serum antioxidant activity in rats. Cecal lactobacilli population was significantly high in all cowpea diet-fed groups compared to control. Modulated serum cholesterol level in cowpea diet-fed rats was accompanied by dietary fiber composition, antioxidant activity in cowpeas and fecal weight, cecal weight and cecal lactobacilli population in rats compared to control. Both processed and raw cowpea-incorporated diets have modulated HFD-induced hypercholesterolemia by modulating serum antioxidative capacity, cholesterol metabolism, and cecal fermentation.

Cowpea: an overview on its nutritional facts and health benefits.

Cowpea (Vigna unguiculata) is a legume consumed as a high-quality plant protein source in many parts of the world. High protein and carbohydrate contents with a relatively low fat content and a complementary amino acid pattern to that of cereal grains make cowpea an important nutritional food in the human diet. Cowpea has gained more attention recently from consumers and researchers worldwide as a result of its exerted health beneficial properties, including anti-diabetic, anti-cancer, anti-hyperlipidemic, anti-inflammatory and anti-hypertensive properties. Among the mechanisms that have been proposed in the prevention of chronic diseases, the most proven are attributed to the presence of compounds such as soluble and insoluble dietary fiber, phytochemicals, and proteins and peptides in cowpea. However, studies on the anti-cancer and anti-inflammatory properties of cowpea have produced conflicting results. Some studies support a protective effect of cowpea on the progression of cancer and inflammation, whereas others did not reveal any. Because there are only a few studies addressing health-related effects of cowpea consumption, further studies in this area are suggested. In addition, despite the reported favorable effects of cowpea on diabetes, hyperlipidemia and hypertension, a long-term epidemiological study investigating the association between cowpea consumption and diabetes, cardiovascular disease and cancer is also recommended. © 2018 Society of Chemical Industry.

Comparative Analysis of Nutritional and Bioactive Properties of Aerial Parts of Snake Gourd (Trichosanthes cucumerina Linn.).

The present investigation was carried out to determine the nutritional and functional properties of T. cucumerina. Water extracts of freeze dried flowers, fruits, and leaves of T. cucumerina were evaluated for their total phenolic content (TPC), total flavonoid content (TFC), antioxidant activity, α-amylase inhibitory activity, and fiber and mineral contents. Antioxidant activity, TPC, and TFC were significantly higher (P ≤ 0.05) in leaves than in flowers and fruits. A significant linear correlation was observed between the TPC, TFC, and antioxidant activities of plant extracts. Although, leaves and flower samples showed a significantly higher (P ≤ 0.05) amylase inhibitory activity than the fruit samples, the overall amylase inhibition was low in all three parts of T. cucumerina. Soluble and insoluble dietary fiber contents were significantly higher (P ≤ 0.05) in fruits than in flowers and leaves. Ca and K contents were significantly higher (P ≤ 0.05) in leaf followed by fruit and flower and Mg, Fe, and Zn contents were significantly higher (P ≤ 0.05) in leaves followed by flowers and fruits. In conclusion, T. cucumerina can be considered as a nourishing food commodity which possesses high nutritional and functional benefits for human health.

A simple microplate-based method for the determination of α-amylase activity using the glucose assay kit (GOD method).

For the first time, a reliable, simple, rapid and high-throughput analytical method for the detection and quantification of α-amylase inhibitory activity using the glucose assay kit was developed. The new method facilitates rapid screening of a large number of samples, reduces labor, time and reagents and is also suitable for kinetic studies. This method is based on the reaction of maltose with glucose oxidase (GOD) and the development of a red quinone. The test is done in microtitre plates with a total volume of 260µL and an assay time of 40min including the pre-incubation steps. The new method is tested for linearity, sensitivity, precision, reproducibility and applicability. The new method is also compared with the most commonly used 3,5-dinitrosalicylic acid (DNSA) method for determining α-amylase activity.

Health-beneficial properties of potato and compounds of interest.

Potatoes have shown promising health-promoting properties in human cell culture, experimental animal and human clinical studies, including antioxidant, hypocholesterolemic, anti-inflammatory, antiobesity, anticancer and antidiabetic effects. Compounds present such as phenolics, fiber, starch and proteins as well as compounds considered antinutritional such as glycoalkaloids, lectins and proteinase inhibitors are believed to contribute to the health benefits of potatoes. However, epidemiological studies exploring the role of potatoes in human health have been inconclusive. Some studies support a protective effect of potato consumption in weight management and diabetes, while other studies demonstrate no effect and a few suggest a negative effect. As there are many biological activities attributed to the compounds present in potato, some of which could be beneficial or detrimental depending on specific circumstances, a long-term study investigating the association between potato consumption and diabetes, obesity, cardiovascular disease and cancer while controlling for fat intake is needed. © 2016 Society of Chemical Industry.

Banana Blossom (Musa acuminate Colla) Incorporated Experimental Diets Modulate Serum Cholesterol and Serum Glucose Level in Wistar Rats Fed with Cholesterol.

Hypocholesterolaemic and hypoglycaemic effect of banana blossom were studied in high-cholesterol fed rats. Experimental groups were fed for 4 weeks, with casein as the basal diet (CN), in comparison with two diets containing 0.5% cholesterol (CD) and 0.5% cholesterol + 21% banana blossom powder (CDB). Serum total cholesterol, non-HDL-cholesterol level, and serum glucose concentrations were lower in CDB fed group compared with CD fed group. Lower serum cholesterol and glucose level (P < 0.05) in CDB fed group were followed by higher faecal weight, caecal weight, caecal Lactobacilli, and Bifidobacteria population in CDB fed group compared to CD diet fed group. Lower serum AST level in banana blossom fed rats showed the reduction in oxidative stress induced by high cholesterol diet. Based on these data, it could be speculated that banana blossom incorporated experimental diets may modulate the hypocholesterolaemic and hypoglycaemic responses in Wistar rats.