Modulation of the gut microbiota and short-chain fatty acid production by gac fruit juice and its fermentation in in vitro colonic fermentation.

This study aimed to investigate the effects of gac fruit juice and its probiotic fermentation (FGJ) utilizing Lactobacillus paracasei on the modulation of the gut microbiota and the production of short-chain fatty acids (SCFAs). We conducted a comparison between FGJ, non-fermented gac juice (GJ), and control samples through in vitro digestion and colonic fermentation using the human gut microbiota derived from fecal inoculum. Our findings revealed that both GJ and FGJ led to an increase in the viability of Lactobacilli, with FGJ exhibiting even higher levels compared to the control. The results from the 16S rDNA amplicon sequencing technique showed that both GJ and FGJ exerted positive impact on the gut microbiota by promoting beneficial bacteria, notably Lactobacillus mucosae and Bacteroides vulgatus. Additionally, both GJ and FGJ significantly elevated the levels of SCFAs, particularly acetic, propionic, and n-butyric acids, as well as lactic acid, in comparison to the control. Notably, FGJ exhibited a more pronounced effect on the gut microbiota compared to GJ. This was evident in its ability to enhance species richness, reduce the Firmicutes to Bacteroidetes (F/B) ratio, promote Akkermansia, and inhibit pathogenic Escherichia coli. Moreover, FGJ displayed enhanced production of SCFAs, especially acetic and lactic acids, in contrast to GJ. Our findings suggest that the probiotic fermentation of gac fruit enhances its functional attributes in promoting a balanced gut microbiota. This beverage demonstrates potential as a functional food with potential advantages for sustaining intestinal health.

Untargeted metabolomics reveal pathways associated with neuroprotective effect of oxyresveratrol in SH-SY5Y cells.

Oxyresveratrol has been documented benefits for neurodegenerative disease. However, the specific molecular mechanisms and pathways involved is currently limited. This study aimed to investigate the potential neuroprotective mechanisms of oxyresveratrol using rotenone-induced human neuroblastoma SH-SY5Y cytotoxicity. Cells were divided into the following groups: control, rotenone, and oxyresveratrol pre-treated before being exposed to rotenone. Cellular assays were performed to investigate neuroprotective effects of oxyresveratrol. The results showed that 20 µM oxyresveratrol was effective in preventing rotenone-induced cell death and decreasing ROS levels in the cells. The alteration of metabolites and pathways involved in the neuroprotective activities of oxyresveratrol were further investigated using LC-QTOF-MS/MS untargeted metabolomics approach. We hypothesized that oxyresveratrol's neuroprotective effects would be associated with neurodegenerative pathways. A total of 294 metabolites were identified. 7,8-dihydrobiopterin exhibited the highest VIP scores (VIP > 3.0; p < 0.05), thus considered a biomarker in this study. Our results demonstrated that pretreatment with oxyresveratrol upregulated the level of 7,8-dihydrobiopterin compared to the positive control. Pathway analysis verified that 7,8-dihydrobiopterin was primarily associated with phenylalanine, tyrosine, and tryptophan metabolism (impact = 1, p < 0.001), serving as essential cofactors for enzymatic function in the dopamine biosynthesis pathway. In conclusion, oxyresveratrol may be benefit for the prevention of neurodegenerative diseases by increasing 7,8-dihydrobiopterin concentration.

Duplex droplet digital PCR (ddPCR) for simultaneous quantification of bovine and porcine gelatin in capsules.

Detection of bovine and porcine in gelatin-based products is important as species fraud and product mislabeling may have a detrimental impact on customers who have health, ethical, and religious concerns about animal products. The duplex droplet digital PCR (ddPCR) assay using double-quenched probes has been developed for quantification and detection of porcine and bovine DNA in gelatin capsules. A DNA mixture derived from gelatin was found to have a limit of detection as low as 0.001 ng/µl for porcine samples and 0.01 ng/µl for bovine samples. DNA from 12 other distinct species was tested with the bovine and porcine probes, showing high specificity for this method. The test was validated using fifty-five commercial supplement and pharmaceutical capsules, of which 17 were positive for bovine and/or porcine DNA. This study shows that the duplex ddPCR is reliable for routine analysis in the identification of bovine and porcine origins for gelatin capsules. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-022-01204-x.

Effects of Maternal Exclusion Diet for Infants Suspected Food Allergy on Fatty Acid Composition in Breast Milk.

Background: Levels of fatty acid (FA) in breast milk (BM) may vary depending on the maternal diet. This study aimed to explore FA composition in BM of lactating women following dietary restrictions due to infant allergic conditions. Materials and Methods: Thai lactating mothers of term infants who were on exclusion diets were recruited. Mature BM was collected before and after a period (at least 2 weeks) of dietary restriction. FA in BM was analyzed by gas chromatography-mass spectrometry. Results: Fifty lactating women 33.7 ± 3.6 years of age were enrolled. Thirty-three percent of the lactating mothers restricted more than eight food items. Most common dietary restriction were cow's milk (88%) and eggs (74%). After the period of dietary exclusion, total polyunsaturated FA showed no significant change, while saturated FA (SFA) declined, and monounsaturated FA (MUFA) increased. A decrease in fat intake was associated with an increase in arachidonic acid (ARA) and docosahexaenoic acid (DHA) content in BM (r = -0.37, r = -0.36; p < 0.05). However, a rise in ARA, eicosapentaenoic acid (EPA), and DHA intake was associated with an increase in linoleic acid and EPA in BM, respectively (r = 0.38, r = 0.55 and r = 0.41; p < 0.05). Infant weight-for-age z-score did not significantly change after the period of maternal dietary exclusion. Conclusion: Maternal exclusion diet resulted in lower SFA and higher MUFA composition in BM. Further study should explore the long-term outcomes of maternal dietary restriction on infant and child health.

Anthocyanin-rich fraction from Thai berries interferes with the key steps of lipid digestion and cholesterol absorption.

Several studies have documented the hypolipidemic effect of anthocyanin-rich plants in vitro and in vivo. The objective of this study was to elucidate the inhibitory activity of anthocyanin-rich fraction from Thai berries against fat digestive enzymes. The ability of Thai berries to bind bile acid, disrupt cholesterol micellization and the cholesterol uptake into Caco-2 cells was also determined. The content of total phenolics, flavonoid and anthocyanin in Prunus domestica L. (TPE), Antidesma bunius (L.) Spreng, Syzygium cumini (L.) Skeels, and Syzygium nervosum A. Cunn. Ex DC was 222.7-283.5 mg gallic acid equivalents, 91.2-184.3 mg catechin equivalents, and 37.9-49.5 mg cyanidin-3-glucoside equivalents/g extract, respectively. The anthocyanin-rich fraction of all extracts inhibited pancreatic lipase and cholesterol esterase with the IC50 values of 90.6-181.7 µg/mL and 288.7-455.0 µg/mL, respectively. Additionally, all extracts could bind primary and secondary bile acids (16.4-36.6%) and reduce the solubility of cholesterol in artificial micelles (53.0-67.6%). Interestingly, TPE was the most potent extract on interfering the key steps of lipid digestion among the tested extracts. In addition, TPE (0.10-0.50 mg/mL) significantly reduced the cholesterol uptake into Caco-2 cells in a concentration-dependent manner. These results demonstrate a new insight into the role of anthocyanin-rich Thai berry extract on interfering the key steps of lipid digestion and absorption.

Inhibitory Effect of Antidesma bunius Fruit Extract on Carbohydrate Digestive Enzymes Activity and Protein Glycation In Vitro.

Antidesma bunius (L.) spreng (Mamao) is widely distributed in Northeastern Thailand. Antidesma bunius has been reported to contain anthocyanins, which possess antioxidant and antihypertensive actions. However, the antidiabetic and antiglycation activity of Antidesma bunius fruit extract has not yet been reported. In this study, we investigated the inhibitory activity of anthocyanin-enriched fraction of Antidesma bunius fruit extract (ABE) against pancreatic α-amylase, intestinal α-glucosidase (maltase and sucrase), protein glycation, as well as antioxidant activity. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) chromatogram revealed that ABE contained phytochemical compounds such as cyanidin-3-glucoside, delphinidin-3-glucoside, ellagic acid, and myricetin-3-galactoside. ABE inhibited intestinal maltase and sucrase activity with the IC50 values of 0.76 ± 0.02 mg/mL and 1.33 ± 0.03 mg/mL, respectively. Furthermore, ABE (0.25 mg/mL) reduced the formation of fluorescent AGEs and the level of Nε-carboxymethyllysine (Nε-CML) in fructose and glucose-induced protein glycation during four weeks of incubation. During the glycation process, the protein carbonyl and ß-amyloid cross structure were decreased by ABE (0.25 mg/mL). In addition, ABE exhibited antioxidant activity through DPPH radical scavenging activity and Trolox equivalent antioxidant capacity (TEAC) with the IC50 values 15.84 ± 0.06 µg/mL and 166.1 ± 2.40 µg/mL, respectively. Meanwhile, ferric reducing antioxidant power (FRAP) showed an EC50 value of 182.22 ± 0.64 µg/mL. The findings suggest that ABE may be a promising agent for inhibiting carbohydrate digestive enzyme activity, reducing monosaccharide-induced protein glycation, and antioxidant activity.

Postmarket Laboratory Surveillance for Forbidden Substances in Halal-Certified Foods in Thailand.

Limited information exists regarding adulteration of Halal-certified food by substances forbidden under Islamic law (Haram substances). This study was conducted using forensic laboratory testing to investigate the prevalence of this type of adulteration. In this large-scale survey of Halal-certified food products randomly collected from markets in Thailand, 4,829 food samples from 10 food groups were tested in the laboratory for four potentially Haram substances: porcine DNA, porcine fatty acids, ethanol, and hydroxyproline (gelatin). No samples were adulterated with porcine DNA or fatty acids. However, 62 samples (approximately 1.3%) were positive for ethanol (>0.5% for non-naturally fermented products and >1% for naturally fermented products). The hydroxyproline concentration in the samples was compared with that of a negative control. Gelatin, as indicated by the presence of hydroxyproline, was the major suspicious substance found in these products. Further investigations are required to determine whether the gelatin is of Halal origin. These results from this first large-scale postmarket surveillance of Halal-certified food products for forbidden substances reveals the important role of forensic laboratory testing for supporting Halal supervision and certification. These findings provide useful information for government agencies seeking to encourage Halal compliance by food enterprises and for Muslim consumers and Halal food importers and exporters.

Moringa Oleifera leaf extract increases plasma antioxidant status associated with reduced plasma malondialdehyde concentration without hypoglycemia in fasting healthy volunteers.

OBJECTIVE: To investigate the effect of Moringa Oleifera leaf extract (MOLE) on plasma glucose concentration and antioxidant status in healthy volunteers. METHODS: A randomized crossover design was used in this study. Healthy volunteers were randomly assigned to receive either 200 mL of warm water (10 cases) or 200 mL of MOLE (500 mg dried extract, 10 cases). Blood samples were drawn at 0, 30, 60, 90, and 120 min for measuring fasting plasma glucose (FPG), ferric reducing ability of plasma (FRAP), Trolox equivalent antioxidant capacity (TEAC) and malondialdehyde (MDA). RESULTS: FPG concentration was not signifificantly different between warm water and MOLE. The consumption of MOLE acutely improved both FRAP and TEAC, with increases after 30 min of 30 µmol/L FeSO4 equivalents and 0.18 µmol/L Trolox equivalents, respectively. The change in MDA level from baseline was signifificantly lowered after the ingestion of MOLE at 30, 60, and 90 min. In addition, FRAP level was negatively correlated with plasma MDA level after an intake of MOLE. CONCLUSION: MOLE increased plasma antioxidant capacity without hypoglycemia in human. The consumption of MOLE may reduce the risk factors associated with chronic degenerative diseases.

Moringa oleifera aqueous leaf extract inhibits reducing monosaccharide-induced protein glycation and oxidation of bovine serum albumin.

Advanced glycation end products (AGEs) play an important factor for pathophysiology of diabetes and its complications. Moringa oleifera is one of the medicinal plants that have anti-hyperglycemic activity. However, anti-glycation property of Moringa oleifera leaf extract on the different types of reducing monosaccharides-induced protein glycation has not been investigated. Therefore, the aim of this study was to examine the protective effect of Moringa oleifera aqueous leaf extract (MOE) on reducing sugars-induced protein glycation and protein oxidation. Total phenolic content of MOE was measured using the Folin-Ciocalteu method. Bovine serum albumin was incubated with 0.5 M of reducing sugars (glucose or fructose) with or without MOE (0.5-2.0 mg/mL) for 1, 2, 3 and 4 weeks. The results found that total phenolic content was 38.56 ± 1.50 mg gallic acid equivalents/g dry extract. The formation of fluorescent and non-fluorescent AGEs [N (ε)-(carboxymethyl) lysine (CML)] and the level of fructosamine were determined to indicate protein glycation, whereas the level of protein carbonyl content and thiol group were examined for protein oxidation. MOE (0.5-2.0 mg/mL) significantly inhibited the formation of fluorescent, N (ε)-CML and markedly decreased fructosamine level (P < 0.05). Moreover, MOE significantly prevented protein oxidation manifested by reducing protein carbonyl and the depletion of protein thiol in a dose-dependent manner (P < 0.05). Thus, the findings indicated that polyphenols containing in MOE have high potential for decreasing protein glycation and protein oxidation that may delay or prevent AGE-related diabetic complications.

The Alterations of Erythrocyte Phospholipids in Type 2 Diabetes Observed after Oral High-Fat Meal Loading: The FTIR Spectroscopic and Mass Spectrometric Studies.

Little is known about the postprandial remodelling of erythrocytes phospholipids (PLs) in type 2 diabetics (T2DM). Therefore, this study aims to compare the alterations of erythrocyte PLs in T2DM to those of healthy subjects after ingestion of a high-fat meal. Eleven T2DM and ten healthy subjects underwent a high-fat meal loading. Erythrocytes were isolated from blood obtained after fasting and 4 h after the meal. Fourier Transform Infrared (FTIR) spectroscopy was initially used to screen erythrocyte PLs by monitoring C-H stretching vibrations. Phosphatidylcholine (PC) molecular species were further investigated by Liquid Chromatography-Electrospray Ionisation-Mass Spectrometry (LC-ESI-MS). For the control group, FTIR revealed postprandial changes in C-H stretching vibrations, particularly of the olefinic band. These findings were supported by LC-ESI-MS data, showing marked changes in PC molecular species, especially of the PC34:1 (where 34 and 1 mean the summed number of carbons and double bonds, respectively). However, similar changes of those were not apparent in the T2DM group. Our results reveal marked postprandial alterations of erythrocyte PC species in healthy subjects whereas only mild alterations are observed in T2DM. The discrepant effects of high-fat meal loading suggest abnormal PC remodelling in the diabetic erythrocyte that may affect its membrane fluidity and integrity.

Androgen disruption and toxicity tests of Butea superba Roxb., a traditional herb used for treatment of erectile dysfunction, in male rats.

OBJECTIVE: To evaluate the effects of the tuberous powder of Butea superba Roxb. (Leguminosae) on blood testosterone and luteinizing hormone (LH), and toxicity in male rats. METHODS: Adult male Wistar rats were orally treated with 0, 10, 100, 150 or 200 mg/kg BW/day of B. superba powder suspension in 0.7 ml distilled water for 90 consecutive days. Blood samples were collected every 30 days and submitted to testosterone and LH analysis. On the 90th day of treatment, blood and the main organs were collected for haematological and histopathological analysis, respectively. RESULTS: The adverse effects found included an increase in spleen relative weight, and increased serum level of alkaline phosphatase (ALP) and aspartate aminotransferase (AST) in rats treated with 150 mg/kg BW/day B. superba powder. At 200 mg/kg BW/day treatment, rats showed significant decreased and increased blood levels of neutrophil and eosinophil, respectively, and a decrease in serum creatinine levels. Serum hormonal analysis revealed a dose-dependent decrease in testosterone, but not LH, in rats treated with 150 and 200 mg/kg BW/day B. superba powder. CONCLUSION: Subchronic treatment of B. superba tuberous powder suspension at high doses in male rats exhibited adverse effects to blood chemistry, haematology, and blood testosterone level. The results of the study should initiate awareness of the possible adverse risk of over-dose consumption of B. superba products for treatment of erectile dysfunction (ED) in mature males.

Alterations of atherogenic low-density lipoproteins and serum fatty acids after 12 week moderate exercise training in sedentary Thai women.

The potential benefit of aerobic exercise upon cardiovascular disease (CVD) through an increasing high-density lipoproteins (HDLs) is acknowledged. However, its effects on low-density lipoproteins (LDLs) and their subpopulations, are unknown in Thailand. Twenty sedentary Thai women undertook a 12-week exercise training program (60% heart rate reserve) comprising 25-minute cycling followed by 10-minute warm-up/cool-down 3 times a week with a group of 20 matched sedentary subjects as control. Triacylglycerols (TGs) and cholesterol (C) of plasma lipoproteins including triacylglycerol-rich lipoproteins (TRLs), large, buoyant LDL (lb-LDL), small, dense LDL (sd-LDL) and HDLs were analyzed while serum fatty acid profiles were also assessed. It was found that plasma TGs, TRL-TGs, sd-LDL-C and sd-LDL-C/lb-LDL-C (S/L) ratio decreased significantly after 12-weeks of exercise to -9%, -8%, -17% and -19% respectively from baseline (p < 0.05). Serum fatty acid profiles remained unchanged. No alteration of any parameters was found in the control group without exercise. These findings suggest that moderate exercise training, even without a change of HDLs, impedes the shift of lb-LDL to more atherogenic sd-LDL, thus possibly preventing cardiovascular disease in healthy, sedentary Thai women.

Major isoflavonoid contents of the phytoestrogen rich-herb Pueraria mirifica in comparison with Pueraria lobata.

Pueraria mirifica tubers collected from 28 out of 76 provinces of Thailand and Pueraria lobata tubers collected from Guangzhou province, China were submitted to HPLC analysis with the established gradient system comprising 1.5% acetic acid and acetonitrile. Five major isoflavonoids, including puerarin, daidzin, genistin, daidzein and genistein, were adopted as authentic standards. P. mirifica tubers showed intra- as well as inter-provincial differences in isoflavonoid and total isoflavonoid contents. The difference in both cases should be mostly influenced by genetic and environmental factors. In comparison with P. lobata, P. mirifica population exhibited differences only with a lower amount of daidzein.