Emmanouil Apostolidis

Effect of Black Tea and Black Tea Pomace Polyphenols on α-Glucosidase and α-Amylase Inhibition, Relevant to Type 2 Diabetes Prevention

This study evaluates the potential mechanism of action and bioactivity of black tea and black tea pomace for type 2 diabetes prevention via inhibition of carbohydrate hydrolyzing enzymes. Black tea leaves were extracted in hot water and black tea pomace was extracted in 70% acetone. The phenolic content of the water extract (WBT) and pomace acetone extracts (AOBT) were 5.77 and 8.9 mg/mL, respectively, both based on the same concentration of solid tea in the extract. The water extract was subjected to C18 extraction and the resulting hydrophobic fraction (HBBT) was further subjected to LH-20 extraction to recover a low molecular weight phenolic enriched fraction (LMW) and a high molecular weight enriched fraction (HMW). The phenolic content of the LMW and HMW fraction were 1.42 and 2.66 mg/mL, respectively. Among water extracts the HMW fraction was most bioactive against α-glucosidase (IC50 = 8.97 μg/mL) followed by HBBT fraction (IC50 = 14.83 μg/mL). However, the HBBT fraction was the most bioactive fraction against α-amylase (IC50 = 0.049 mg/mL). The black tea pomace (AOBT) had significant α-glucosidase inhibitory activity (IC50 = 14.72 μg/mL) but lower α-amylase inhibitory activity (IC50 = 0.21 mg/mL). The phenolic profiles for LMW and HMW fractions were evaluated using HPLC and the differences between the two profiles were identified. Further research is underway to identify and evaluate the phenolic compounds that are present in the HMW fraction. Our findings suggest that black tea and black tea pomace has potential for carbohydrate hydrolyzing enzyme inhibition and this activity depends on high molecular weight phenolic compounds.

In Vitro and In Vivo Antihyperglycemic Effect of 2 Amadori Rearrangement Compounds, Arginyl-Fructose and Arginyl-Fructosyl-Glucose

During the heat processing of raw ginseng to produce red ginseng, amino acid derivatives such as arginyl-fructose (AF) and arginyl-fructosyl-glucose (AFG) are formed at high levels, through amadori rearrangement, the early step of Maillard reaction, from arginine and glucose or maltose, respectively. However, very limited information is available about the effect of the structural difference between AF and AFG on various biological activities. This is the first report of the mode of action and effect of AF and AFG on the type 2 diabetes management related inhibition of postprandial hyperglycemia in vitro and in animal model. In our previous study, standards AF and AFG were chemically synthesized and in this study their inhibitory activities against rat intestinal α-glucosidases and porcine pancreatic α-amylase were investigated in vitro. The IC(50) value of the in vitro inhibitory activity of AF and AFG on rat intestinal sucrase was high and in similar levels (6.40 and 6.20 mM, respectively). Additionally, a mild pancreatic α-amylase inhibitory activity was observed, with IC(50) values 36.30 and 37.60 mM for AF and AFG, respectively. The effect of AF and AFG on the postprandial blood glucose increase after meal was investigated in Sprague Dawley rats fed on starch or sucrose meals. Both amadori compounds significantly reduced the postprandial blood glucose levels after starch or sucrose loading. These results indicate that AF and AFG, Maillard reaction products, may have antidiabetic effect by suppressing carbohydrate absorption in the gastrointestinal level, and thereby reducing the postprandial increase of blood glucose.

The reduction effect of low molecular weight chitosan oligosaccharide (GO2KA1) on postprandial blood glucose levels in healthy individuals

The effects of chitosan-oligosaccharide (GO2KA1) on postprandial blood glucose levels in adults with normal blood glucose levels were investigated. Postprandial blood glucose levels were measured at 30, 60, 90, and 120 min after sucrose administration with and without 500 mg of GO2KA1. GO2KA1 administration reduced the area under the blood glucose-time curve (AUC) and the blood glucose peak (Cmax) values while the time of peak plasma concentration of blood glucose (Tmax) value was significantly (p<0.05) increased, compared to controls. GO2KA1 reduced postprandial blood glucose level increases via slower absorption of glucose in the small intestine based on carbohydrate hydrolyzing enzyme inhibition.

Seasonal influence on phenolic-mediated antihyperglycemic properties of Canadian sugar and red maple leaves using in vitro assay models

Red and sugar maple leaves collected in the summer and fall from Canada, were evaluated for phenolic content, antioxidant, α-glucosidase, and α-amylase inhibitory activities variation. The phenolic contents of summer red maple leaves (RML-S) and summer sugar maple leaves (SML-S) were higher than red and sugar maple leaves collected in fall (RML-F and SML-F, respectively). HPLC analyses showed differences in phenolic compounds present in the SML samples compared to the RML samples. The extracts were assayed for yeast and rat α-glucosidase inhibitory activities. Both results showed that SML-S extracts had the highest inhibitory activity which could possibly be attributed to the unique phenolics present therein. Milder effects were observed in terms of α-amylase inhibitory activity, with RML-F having the highest inhibitory activity. These results suggest that maple tree leaf extracts may have potential for phenolic-mediated α-glucosidase inhibition, relevant to type 2 diabetes management, with SML-S extract having the highest bioactivity.

Recovery of Bioactive Peptides and Omega-3 Fatty Acids-Containing Phospholipids from Squid Processing By-Product Hydrolysate

ABSTRACT The present study examined whether bioactive peptides and omega-3 fatty acids-containing phopholipids could be recovered from squid processing by-product (SPB) hydrolysate. The hydrolysate was produced at 55°C for varying times with endogenous proteases and centrifuged to yield peptides-containing supernatant and phospholipids-containing precipitate. The supernatant showed angiotensin I-converting enzyme (ACE) inhibitory activity with half maximal inhibitory concentration (IC50) values decreasing with hydrolysis time from 2.11 mg (0 h) to 1.71 mg (1 h), 1.38 mg (1.5 h), and 1.34 mg (2 h). Two-hour squid hydrolysate was further fractionated to isolate the most active fraction whose molecular weight was found to be below 10 kDa with IC50 of 0.32 mg. The phospholipids-containing precipitate (45.6 g/100 g oil) was analyzed for a fatty acid profile, with the levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) being 16.86 and 29.24 g/100 g oil, respectively. The ACE inhibition by hydrolysate peptides and omega-3 fatty acid recovery support the nutraceutical potential of the SPB hydrolysate.

Antidiabetic effect of chitosan oligosaccharide (GO2KA1) is mediated via inhibition of intestinal alpha‐glucosidase and glucose transporters and PPARγ expression

We have previously reported that administration of low molecular weight chitosan oligosaccharide (GO2KA1) significantly suppressed postprandial blood glucose rise with increased plasma adiponectin and HbA1c levels in animals and humans. However, the cellular mechanisms whereby GO2KA1 exerts antihyperglycemic effects still remain to be determined. Using intestinal Caco-2 cells and 3T3-L1 cells, here we show that GO2KA1 has dual modes of antidiabetic action by (1) inhibiting intestinal α-glucosidase as well as glucose transporters SGLT1 and GLUT2 that were distinct from the acarbose effect; (2) enhancing adipocyte differentiation, PPARγ expression and its target genes, such as FABP4, adiponectin, and GLUT4, whereas the effects were abolished by co-treatment with BADGE, a PPARγ antagonist. Moreover, GO2KA1 significantly increased glucose uptake, which was reduced in the presence of BADGE. Our data show that GO2KA1 may prevent hyperglycemia by inhibiting intestinal glucose digestion and transport and also enhance glucose uptake, at least in part, by upregulating adiponectin expression through PPARγ in adipocytes. These findings may provide potential molecular modes of action for the antidiabetic effects of chitosan oligosaccharide observed in clinical and animal studies.

Evaluation of Phenolic Phytochemical Enriched Commercial Plant Extracts on the In Vitro Inhibition of α-Glucosidase

Green tea (GT), cranberry (CR), and tart cherry extracts were evaluated for their ability to inhibit yeast α-glucosidase, relevant to glucose uptake. The total phenolic content (TPC), antioxidant activity, and in vitro inhibitory activity of yeast α-glucosidase were examined for the extracts in the present study. GT had higher TPC and antioxidant activity, but CR demonstrated a greater α-glucosidase inhibitory activity, on phenolic basis. CR was fractionated using LH-20 column chromatography into two fractions: 30% methanol (CME) and 70% acetone (CAE). TPC, antioxidant activity, and yeast α-glucosidase inhibitory activity were determined for the fractions. CAE had a greater TPC and antioxidant activity than CME, but the two fractions had a synergistic effect when inhibiting yeast α-glucosidase. Our findings suggest that CR has the greatest potential to possibly manage post-prandial blood glucose levels via the inhibition of α-glucosidase, and that the effect is through synergistic activity of the extracts phenolic compounds.

Effect of supplementation of low-molecular-weight chitosan oligosaccharide, GO2KA1, on postprandial blood glucose levels in healthy individuals following bread consumption

The effect of chitosan oligosaccharide (GO2KA1) administration on postprandial blood glucose levels of subjects with normal blood glucose levels was evaluated following bread consumption. Postprandial blood glucose levels were determined for 2 h after bread ingestion with or without 500 mg of GO2KA1. GO2KA1 significantly lowered the mean, maximum, and minimum levels of postprandial blood glucose at 30 min after the meal. Postprandial blood glucose levels were decreased by about 25% (from 155.11±13.06 to 138.50±13.59, p<0.01) at 30 min when compared to control. Furthermore, we observed that the area under the concentration-time curve (AUCt) was decreased by about 6% (from 255.46±15.43 to 240.15±14.22, p<0.05) and the peak concentration of blood glucose (Cmax) was decreased by about 11% (from 157.94±10.90 to 140.61±12.52, p<0.01) when compared to control. However, postprandial the time to reach Cmax (Tmax) levels were the same as those found in control. Our findings suggest that GO2KA1 limits the increase in postprandial blood glucose levels following bread consumption.

Comparison of the Antimicrobial and Antioxidant Activities of Selected Wheat Varieties

Antibacterial and antioxidant activities of wheat seed ethyl acetate extracts for Jokyoung (JK), Dark northern spring (DNS), Keumkang (KK), Woori (WR), and Winter wheat (WW) were investigated. Antibacterial activities were evaluated in vitro against the common food and cosmetic industry contaminants Escherichia coli, Salmonella typhimurium, and Staphylococcus aureus using well diffusion assays. WW had the highest inhibitory activity against all tested strains, with S. aureus being the most sensitive strain. The minimum inhibitory concentration (MIC) values of WW and WR against S. aureus were 0.50 and 1.25 mg/mL, respectively. The 2,6-dimethoxy-1,4- benzoquinone (DMBQ) content was measured using HPLC. The antibacterial activities of wheat seed extracts were correlated with the total phenolic contents (Pearson’s correlation coefficient=0.994), with the ABTS radical scavenging activity (0.978), and with the DMBQ content (0.968). WW and WR have potential for use as natural antimicrobials for prevention of food and cosmetics spoilage.