Identification of MATE Family and Characterization of GmMATE13 and GmMATE75 in Soybean's Response to Aluminum Stress.

The multidrug and toxic compound extrusion (MATE) proteins are coding by a secondary transporter gene family, and have been identified to participate in the modulation of organic acid exudation for aluminum (Al) resistance. The soybean variety Glycine max "Tamba" (TBS) exhibits high Al tolerance. The expression patterns of MATE genes in response to Al stress in TBS and their specific functions in the context of Al stress remain elusive. In this study, 124 MATE genes were identified from the soybean genome. The RNA-Seq results revealed significant upregulation of GmMATE13 and GmMATE75 in TBS upon exposure to high-dose Al3+ treatment and both genes demonstrated sequence homology to citrate transporters of other plants. Subcellular localization showed that both proteins were located in the cell membrane. Transgenic complementation experiments of Arabidopsis mutants, atmate, with GmMATE13 or GmMATE75 genes enhanced the Al tolerance of the plant due to citrate secretion. Taken together, this study identified GmMATE13 and GmMATE75 as citrate transporter genes in TBS, which could improve citrate secretion and enhance Al tolerance. Our findings provide genetic resources for the development of plant varieties that are resistant to Al toxicity.

Fecal metabonomics combined with 16S rDNA sequencing to analyze the changes of gut microbiota in rats fed with different protein source diets.

PURPOSE: When blended, animal and plant proteins can complement each other in terms of amino acid composition and release time. In this study, we investigated whether the blended protein diet has a better feeding effect than the single protein diet, and to reveal the differences in growth and intestinal microbiota composition caused by the blended protein diet. METHODS: Forty Sprague Dawley (SD) rats received diets with different protein sources, including casein (C), whey protein (WP), black soybean protein (BSP), and black soybean-whey blended protein (BS-WP), for eight weeks. To investigate the effects of blended protein supplement on gut microbiota and metabolites, we performed a high throughput 16S rDNA sequencing and fecal metabolomics profiling. In addition, we determined growth and serum biochemical indices, and conducted intestinal morphology analyses. RESULTS: Compared to those in the BSP and WP groups, the daily body weight gain and feed conversion efficiency increased in the BS-WP group. Serum biochemical indices indicated that the protein utilization efficiency of the WP and BS-WP groups was relatively high, and the BS-WP blended protein diet improved the protein adoption rate. The BS-WP blended protein diet also improved intestinal tissue morphology and promoted intestinal villi development compared to the single protein diets. Furthermore, dietary protein altered the composition of gut microbiota, the gut microbial diversity of rats fed with the BS-WP diet was significantly (P < 0.05) higher than that of the other groups. The difference in dietary protein corresponded with an alteration of fecal amino acids and their metabolites, and tryptophan and tyrosine metabolism were the key mechanisms leading to the changes in fecal microbial composition. CONCLUSION: Dietary protein sources played an important role in the growth and development of rats by influencing intestinal metabolism and microbial composition. The BS-WP blended protein diet was more conducive to nutrient absorption than the single protein diet. Furthermore, blended protein increased the diversity of intestinal microbes and aided the establishment of intestinal barrier function.

Pancreatin-induced liberation of starch/cyanidin 3-O-glucoside complexes from rice cooked with black soybean that exhibit slow hydrolysis.

Cyanidin 3-O-glucoside (C3G), which has various health-promoting functions, is contained in black soybean (BSB). In Japan and Korea, BSB is cooked with rice and the cooked rice appears purplish in colour. In this study, BSB was cooked with glutinous rice, non-glutinous rice, and high-amylose rice. The amount of C3G detected in high-amylose rice was greater than that detected in glutinous rice, suggesting that C3G combined more efficiently with amylose than with amylopectin. Pancreatin induced the liberation of starch/C3G complexes from the purplish cooked rice, and rate of the liberation was in the following order; glutinous rice < non-glutinous rice < high-amylose rice. The amylose/C3G complexes liberated from high-amylose rice was hydrolysed slowly, while the amylopectin/C3G complexes liberated from glutinous rice were hydrolysed into smaller amylopectin/C3G complexes that were difficult to further hydrolysis. Thus, C3G may be useful for preparing foods whose starch hydrolysis is slow.

Extract from Black Soybean Cultivar A63 Extract Ameliorates Atopic Dermatitis-like Skin Inflammation in an Oxazolone-Induced Murine Model.

Black soybean has been used in traditional medicine to treat inflammatory diseases, cancer, and diabetes and as a nutritional source since ancient times. We found that Korean black soybean cultivar A63 has more cyanidin-3-O-glucoside, (C3G), procyanidin B2 (PB2), and epicatechin (EPC) contents than other cultivars and has beneficial effects on cell viability and anti-oxidation. Given the higher concentration of anthocyanidins and their strong anti-oxidant activity, we predicted that A63 extract could relieve inflammatory disease symptoms, including those of atopic dermatitis (AD). Here, we evaluated the anti-AD activity of A63 extract in an oxazolone (OXA)-induced mouse model. A63 extract treatment significantly reduced epidermal thickness and inflammatory cell infiltration, downregulated the expression of AD gene markers, including Interleukin (IL)-4 and IL-5, and restored damaged skin barrier tissues. Furthermore, A63 extract influenced the activation of the signal transducer and activator of transcription (STAT) 3 and STAT6, extracellular regulatory kinase (ERK), and c-Jun N-terminal kinase (JNK) signaling pathways, which play a crucial role in the development of AD. Altogether, our results suggest that A63 can ameliorate AD-like skin inflammation by inhibiting inflammatory cytokine production and STAT3/6 and Mitogen-activated protein kinase (MAPK) signaling and restoring skin barrier function.

Experimental and theoretical research on the effect of coupling heat and pH on the structure and antioxidant activity of cyanidin-3-O-glucoside from black soybean coat.

BACKGROUND: Since anthocyanin has good coloration and antioxidant properties, many studies have focused on exploring the stability and antioxidant activity of anthocyanin. The objective of this work was to study effects of pH and temperature on the bioactivity of cyanidin-3-O-glucoside (C3G) and ultra-performance liquid chromatography-photodiode array-electrospray ionization-quadrupole-time-of-flight mass spectrometry (UPLC-PDA-ESI-Q-TOF-MS) and density functional theory (DFT) were used to explain the mechanism of structural transformation of C3G affecting their bioactivity at the molecular level. RESULTS: During the heating process at pH 2.2 to 7.0，the flavylium cation content of C3G decreased from 92.71% to 51.64% and the chalcone content increased from 7.29% to 30.61%. The quinoidal base and first discovery of the degradation product of the C3G, 1-(3,4-dihydroxy-phenyl)-2-(3, 4,5-trihydroxy-6-hydroxymethyl-tetrahydro-pyran-2-yloxy)-ethanone, were only detected in heated samples at pH 7.0. DFT revealed the antioxidant mechanism was mediated by sequential proton loss electron transfer and the antioxidant activity of C3G in pH 5.0 and 7.0 environments was higher than that in the pH 2.2 environment. CONCLUSIONS: The results revealed the thermal degradation products of C3G included catechin, 3,4-dihydroxybenzoic acid, 2,4,6-trihydroxy-benzaldehyde and 1-(3,4-Dihydroxy-phenyl)-2-(3,4,5-trihydroxy-6-hydroxymethyl-tetrahydro-pyran-2-yloxy)-ethanone. C3G had higher antioxidant activity in weakly acidic to near-neutral environments and the reactive sites were most likely at the 4'-OH and 5-OH sites. © 2021 Society of Chemical Industry.

Fermentation of black soybean with Bacillus spp. for the production of kinema: changes in antioxidant potential on fermentation and gastrointestinal digestion.

Black soybean was fermented with four different potential Bacillus spp., including Bacillus licheniformis K1G, Bacillus subtilis K2B, Bacillus amyloliquefaciens K2G and Bacillus subtilis K2M, isolated from kinema, a traditionally fermented soybean product of Sikkim. Enhancement of antioxidant activity was observed with DPPH radical scavenging activity, reducing power potential and total antioxidant activity in methanolic as well as water extracts. Overall antioxidant activities were found to be higher in fermented black soybean in comparison to yellow soybean, showing its potential for production of kinema. Further, black soybean fermented using different starter was subjected to gastrointestinal digestion using pepsin and pancreatin. Upon gastrointestinal digestion of fermented black soybean changes in antioxidant activity was observed that was found to be reliant on the species and strains applied for fermentation as starter culture. Among different starters used for fermentation, black soybean fermented using B. subtilis K2M had higher DPPH radical scavenging and reducing power activity on gastrointestinal digestion. This study concludes that B. subtilis K2M can be applied for fermentation of black soybean for production of kinema as well as bioactive protein hydrolysates. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13197-021-05144-y.

Black soybean improves the vascular function through an increase in nitric oxide and a decrease in oxidative stress in healthy women.

Vascular dysfunction and injurious stimuli such as oxidative stress is closely related to the risk of cardiovascular diseases (CVD). Dietary polyphenols is reported to exert the beneficial effects on reducing the risk of CVD. Black soybean is rich in polyphenols, including isoflavones, anthocyanidins and flavan-3-ols, and its prevention effects on CVD risk were reported in the animal experiments. In this study, we investigated the effect of black soybean consumption on the vascular function and oxidative stress associating with the polyphenol concentrations in healthy women. Lowered vascular age was observed in 33 out of 44 volunteers who completed the 8-week trial. It was observed that improvement of the vascular stiffness, increasing in the urinary NO2 and NO3 level, and decreasing in the oxidative stress markers, 8-hydroxy-2'-deoxyguanosine, hexanoyl-lysine and myeloperoxidase. In addition, concentration of 12 polyphenols in black soybean increased in the plasma and urine. Increased concentration of polyphenols would be involved in the decreased oxidative stress. Thus, black soybean consumption improved the vascular function through an increase in nitric oxide and a decrease in oxidative stress accompanied by increasing the polyphenol concentrations in healthy women.

Effects of Modified Processing Methods on Structural Changes of Black Soybean Protein Isolate.

To explore better methods of natural protein modification for black soybean, comparisons among the effects of different modified methods on structural changes of the modified products of black soybean protein isolate (BSPI) were carried out in this study. The modified products used in this study included enzymatic crossing-link black soybean protein isolate (ECBSPI), wet heating treatment glycosylation black soybean protein isolate (WHTGBSPI) and especially enzymatic glycosylation black soybean protein isolate catalyzed by transglutaminase (EGBSPI). The effects of the modification methods on structural changes were analyzed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), amino acid content and circular dichroism (CD) analysis. Moreover, the processing properties changes caused by structural changes of BSPI were detected by thermogravimetric analysis, particle size analysis, zeta-potential, surface hydrophobicity, solubility, emulsification, gelation, and rheological properties. The results show that the modified BSPI products were protein polymers, and among them, EGBSP and WHTGBSPI are covalently bonded glycation products. Products modified by Maillard reactions and transglutaminase (TG) display partly destroyed α-helix and ß-sheet structures that form more open secondary BSPI structures. For ECBSPI, the proportion of irregular crimp structure reduces to form a high order secondary structure. All the modified products form fine aggregations in dispersion, except WHTGBSPI has most negative zeta-potential and least molecular stability due to the hydrophobic amino acids embedded in the protein molecules. The zeta-potentials of BSPI, ECBSPI, WHTGBSPI and EGBSPI are respectively -21.5, -23.8, -18.1 and -20.2 mV. The surface hydrophobicity of EGBSPI (5.07 ± 0.07) and WHTGBSPI (7.02 ± 0.05) decrease, while the surface hydrophobicity of ECBSPI (19.5 ± 0.06) increases. The solubility and rheological properties of EGBSPI, ECBSPI and WHTGBSPI after modification are all better than those of BSPI, especially EGBSPI. Emulsification of EGBSPI and WHTGBSPI increase (by 24.5% and 12.2%, respectively) while ECBSPI decrease (by 17.0), and there is similar emulsion stability trend. Moreover, the properties of ECBSPI increase except cohesiveness compared to BSPI. In conclusion, as a safe and efficient method for natural protein modification, enzymatic glycosylation catalyzed by TG has great potential in improving food processing characteristics.

Microencapsulation of anthocyanin-rich black soybean coat extract by spray drying using maltodextrin, gum Arabic and skimmed milk powder.

Black soybean coat is insufficiently valorised food production waste rich in anthocyanins. The goal of the study was to examine physicochemical properties of spray dried extract of black soybean coat in regard to carrier materials: maltodextrin, gum Arabic, and skimmed milk powder. Maltodextrin and gum Arabic-based microparticles were spherical and non-porous while skimmed milk powder-based were irregularly shaped. Low water activity of microparticles (0.31-0.33), good powders characteristics, high solubility (80.3-94.3%) and encapsulation yields (63.7-77.0%) were determined. All microparticles exhibited significant antioxidant capacity (243-386 µmolTE/g), good colour stability after three months of storage and antimicrobial activity. High content of total anthocyanins, with cyanidin-3-glucoside as predominant, were achieved. In vitro release of anthocyanins from microparticles was sustained, particularly from gum Arabic-based. These findings suggest that proposed simple eco-friendly extraction and microencapsulation procedures could serve as valuable tools for valorisation and conversion of black soybean coat into highly functional and stable food colourant.

Black soybean extract reduces fatty acid contents in subcutaneous, but not in visceral adipose triglyceride in high-fat fed rats.

It is known that black soybean (BS) extract, rich in polyphenols, has beneficial effects against obesity, inflammation and insulin resistance. However, detailed effects of BS on lipid metabolism have not been documented well. In the present study, we compared fatty acid composition in visceral and subcutaneous adipose tissues of high-fat fed (HFF) rats and BS administered HFF rats. Black soybean administration for 6 weeks influenced neither body nor adipose tissue weights, blood glucose, plasma insulin levels, or insulin sensitivity. However, BS reduced several saturated (C14:0 and C16:0), monounsaturated (C14:1n-5 and C18:1n-9) and n-6 polyunsaturated (C18:2n-6, C20:3n-6, C20:4n-6 and C22:4n-6) fatty acid contents in subcutaneous fat without any change in n-3 polyunsaturated fatty acid contents. No such effect was observed in fatty acid composition in visceral fat. Long-chain fatty acids are involved in regulation of inflammation. Therefore, those reduced fatty acids may be linked to the effects on suppressing inflammation.

Wide genetic variation in phenolic compound content of seed coats among black soybean cultivars.

Black soybeans have been used as a food source and also in traditional medicine because their seed coats contain natural phenolic compounds such as proanthocyanidin and anthocyanin. The objective of this research is to reveal the genetic variation in the phenolic compound contents (PCCs) of seed coats in 227 black soybean cultivars, most of which were Japanese landraces and cultivars. Total phenolics were extracted from seed coats using an acidic acetone reagent and the proanthocyanidin content, monomeric anthocyanin content, total flavonoids content, total phenolics content, and radical scavenging activity were measured. The cultivars showed wide genetic variation in PCCs. Each of the contents was highly correlated with one another, and was closely associated with radical scavenging activity. PCCs were also moderately associated by flowering date but not associated by seed weight. Cultivars with purple flowers had a tendency to produce higher PCCs compared with cultivars with white flowers, suggesting that the W1 locus for flower color can affect phenolic compound composition and content. Our results suggest that developing black soybean cultivars with high functional phenolic compounds activity is feasible.

Effect of thermal treatments on anti-nutritional factors and antioxidant capabilities in yellow soybeans and green-cotyledon small black soybeans.

BACKGROUND: The effects of different thermal processes on the anti-nutritional factors (tannins, phytic acids and trypsin inhibitors) and antioxidant capacity of yellow soybeans (YS) and green-cotyledon small black soybeans (GCSBS) were investigated. The soybean samples were subjected to roasting, microwaving and boiling as the thermal treatments. RESULTS: Sixty-minute boiling was most effective for eliminating the tannins in both YS and GCSBS, and also resulted in the maximal reduction of phytic acids. Trypsin inhibitors were most effectively reduced (76.92-97.91%) by microwaving, and boiling was least favourable for retaining total phenolic content. The total flavonoid content was significantly (P < 0.05) decreased by all thermal processes, and particularly by employing 60-min boiling. The total anthocyanin content of GCSBS was higher than that of YS, but significantly declined upon boiling (P < 0.05). Roasting did not significantly affect the DPPH free radical scavenging activity of GCSBS (P > 0.05). Microwaving increased the FRAP (ferric reducing antioxidant power) of GCSBS up to 6.07%. CONCLUSION: Our raw GCSBS possesses much better antioxidant capacity than raw YS. Microwaving resulted in a substantial reduction of the trypsin inhibitors of our soybean samples. Dry thermal treatments caused slight increase of FRAP in GCSBS.

Identification of anthocyanins in black soybean (Glycine max (L.) Merr.) varieties.

Anthocyanin pigments in black soybean (Glycine max (L.) Merr.) varieties as Tawonkong (TW) and Geomjeongkong-2 (G2) were identified to evaluate their potentials as nutritional function, natural colorant or functional foods. Anthocyanin extraction was conducted with acidified methanol with 0.1 M HCl (85:15, v/v). Identification of anthocyanin was conducted by comparison with purified standards by HPLC and mass spectrometry (MS). G2 showed six different types of pigments by HPLC, whereas TW showed seven pigments. Three major anthocyanins (peaks 1, 3, 4) were detected in both varieties and peak 1 was characterized as delphinidin-3-O-ß-D-glucoside, peak 3 as cyanidin-3-O-ß-D-glucoside (C3G), and peak 4 as petunidin-3-O-ß-D-glucoside by comparison of chromatographic properties with authentic standards and MS. Minor peaks 5, 6 and 7 in TW were tentatively identified as pelargonidin-3-O-glucoside, pelargonidin-3-O-(6″-malonylglucoside) and cyanidin on the basis of MS. MS with major ions at 287 and 449 of peak 2 were exactly same as those of peak 3 meaning that peak 2 has cyanidin and other hexose different from glucose. After acid hydrolysis of fractioned peak 2, HPLC showed the hexose as galactose, and peak 2 was identified as C3Glactose. The most abundant anthocyanin in black soybean (Glycine max (L.) Merr.) was C3G and G2 showed the higher amount of total anthocyanins than TW (p < 0.001).

Comparison of the anti-diabetic effects of various grain and legume extracts in high-fat diet and streptozotocin-nicotinamide-induced diabetic rats.

The anti-diabetic properties of whole groats and dietary fibers from various grains and legumes are well known. Nevertheless, studies on the anti-diabetic effects of their extracts are limited, and it is difficult to compare their efficacy. This study investigated the anti-diabetic potential of ethanol extracts from oats (OE), sorghum (SE), foxtail millet (FE), proso millet (PE), adzuki bean (AE), and black soybean (BE) in a high-fat diet and streptozotocin-nicotinamide-induced diabetic rat models. The extracts, obtained using 99.9 % ethanol, were orally administered to diabetic rats for four weeks. Various parameters were evaluated, including fasting blood glucose levels, glucose tolerance, insulin sensitivity, serum insulin levels, and pancreas histological analysis. OE and SE effectively reduced fasting blood glucose levels and the area under the curve (AUC) in the oral glucose tolerance test. Only OE significantly decreased the AUC in the insulin tolerance test and increased insulin concentration and homeostatic model assessment of the ß-cell function index, indicating improved insulin sensitivity and ß-cell function. Histological and immunohistochemical analysis of the pancreas supported these findings, demonstrating that OE protected against pancreatic cell damage. In contrast, FE, PE, AE, and BE did not have a significant effect on diabetes-related parameters. These findings identify OE as the most promising natural intervention for diabetes management.

Black Soybean Seed Coat Extract Improves Endothelial Function and Upregulates Oxidative Stress Marker Expression in Healthy Volunteers by Stimulating Nitric Oxide Production in Endothelial Cells.

Black soybean seed coat extract (BE) contains multiple bioactive polyphenols, including flavan-3-ols and anthocyanins. BE improves endothelial function; however, it is unclear whether BE protects endothelial cells from senescence. In this study, we examined the effects of BE on endothelial cell senescence and vascular function in healthy individuals. High concentrations of glucose were used to induce senescence in bovine aortic endothelial cells incubated with BE. Senescence, vascular function, and oxidative stress markers were measured. Incubation with BE remarkably inhibited senescence-associated ß-galactosidase and lactate dehydrogenase activities and dose dependently reduced intracellular reactive oxygen species levels in bovine aortic endothelial cells. BE treatment increased the levels of endothelial nitric oxide synthase (eNOS) mRNA and endothelial nitric oxide (NO) metabolites and increased the mRNA expression of klotho, a gene associated with an antiaging phenotype. To examine the effects of BE in humans, we conducted a clinical study using the second derivative of the fingertip photoplethysmogram to investigate vascular function and aging in 24 healthy volunteers. The participants consumed BE supplements (100 mg/day) or a placebo for 2 weeks. When compared with the placebo group, the BE group showed considerably improved vascular function, NO metabolite levels, and oxidative stress. These results suggest that BE supplementation improves endothelial function, possibly through antioxidant activity and NO production, and may consequently reduce the cardiovascular risk associated with aging. BE supplementation may be an effective and safe approach to reduce the risk of atherosclerosis and cardiovascular disease; however, additional studies investigating chronic vascular inflammation are needed.

Enrichment of Flavonoids in Short-Germinated Black Soybeans (Glycine max L.) Induced by Slight Acid Treatment.

Exogenous abiotic stimulant treatments are a straightforward and effective method for enhancing secondary metabolites in plants. In this study, the response surface optimization method was used to optimize the conditions for enriching flavonoids in short-germinated black soybeans under a slight acid treatment, and the mechanism of flavonoid accumulation during black soybean germination was explored. The results show that the use of a 126.2 mM citric acid-sodium citrate buffer (pH 5.10) as a slight acid treatment resulted in the highest flavonoid content when the black soybeans were germinated for 24 h. Under these conditions, the isoflavonoid (glycitin, daidzein, and genistein) increased significantly, and the flavonoid content reached 2.32 mg/g FW. The microacidified germination treatment significantly increased the activities and relative gene expression levels of key enzymes involved in flavonoid metabolism (4-coumarate-CoA ligase and cinnamic acid 4-hydroxylase, etc.). However, the slight acid treatment inhibited the growth of the black soybeans and caused damage to their cells. This was evidenced by significantly higher levels of malondialdehyde, superoxide anion, and hydrogen peroxide compared to the control group. Furthermore, the antioxidant system in the short-germinated soybeans was activated by the slight acid treatment, leading to a significant increase in the activities and relative gene expression levels of catalase and peroxidase. The results above show that a slight acid treatment was beneficial in inducing the accumulation of flavonoids during the growth of black soybean sprouts. This lays a technical foundation for producing black soybean products that are rich in flavonoids.

Effects of preheat treatment and syringic acid modification on the structure, functional properties, and stability of black soybean protein isolate.

This study investigated preheated (25-100°C) black soybean protein isolate (BSPI) conjugated with syringic acid (SA) (25 and 50 µmol/g protein) under alkaline conditions, focusing on the structure, functional properties, and storage stability. The results revealed that the SA binding equivalent and binding rate on BSPI increased continuously as the preheat temperature increased. Additionally, preheating positively impacted the surface hydrophobicity (H0) of BSPI, with further enhancement observed upon SA binding. Preheating and SA binding altered the secondary and tertiary structure of BSPI, resulting in protein unfolding and increased molecular flexibility. The improvement in BSPI functional properties was closely associated with both preheating temperature and SA binding. Specifically, preheating decreased the solubility of BSPI but enhanced the emulsifying activity index (EAI) and foaming capacity (FC) of BSPI. Conversely, SA binding increased the solubility of BSPI with an accompanying increase in EAI, FC, foaming stability, and antioxidant activity. Notably, the BSPI100-SA50 exhibited the most significant improvement in functional properties, particularly in solubility, emulsifying, and foaming attributes. Moreover, the BSPI-SA conjugates demonstrated good stability of SA during storage, which positively correlated with the preheating temperature. This study proposes a novel BSPI-SA conjugate with enhanced essential functional properties, underscoring the potential of preheated BSPI-SA conjugates to improve SA storage stability. PRACTICAL APPLICATION: Preheated BSPI-SA conjugates can be used as functional ingredients in food or health products. In addition, preheated BSPI shows potential as a candidate for encapsulating and delivering hydrophobic bioactive compounds.

A Standardized Extract of Green-Cotyledon Small Black Soybean (EYESOY®) Ameliorates Dry Eye Syndrome in an Animal Model.

PURPOSE: Dry eye syndrome is a common ocular disease that causes morbidity, high healthcare burden, and decreased quality of life. In this study, we evaluated the beneficial effects of a standardized extract of small black soybean (EYESOY®) in a benzalkonium chloride (BAC)-induced murine model of dry eye. METHODS: Experimental dry eye was induced by instillation of 0.02% BAC on the right eye of the Sprague-Dawley rats. Saline solution or EYESOY were administered orally every day for 8 weeks. RESULTS: EYESOY significantly improved tear volume in the cornea compared with that in the BAC group. Moreover, EYESOY inhibited damage to the corneal epithelial cells and lacrimal glands by suppressing the oxidative and inflammatory responses in a mouse dry eye model. It also increased the goblet cell density and mucin integrity in the conjunctiva. CONCLUSIONS: Our results suggest that EYESOY has the potential to alleviate dry eye syndrome.

Anti-Inflammatory Effects of Anthocyanin-Enriched Black Soybean Seed Coat (BSSC) Crude Extract on LPS-Induced Acute Liver Injury in Mice.

Soybeans rank among the top five globally produced crops. Black soybeans contain anthocyanins in their seed coat, offering strong antioxidant and anti-inflammatory benefits. This study explores the protective effects of black soybean seed coat (BSSC) against acute liver injury (ALI) in mice. Mice pretreated with BSSC crude extract showed reduced liver damage, inflammation, and apoptosis. High doses (300 mg/kg) of the extract decreased levels of proinflammatory cytokines (IL-6, IFN-γ) and increased levels of anti-inflammatory ones (IL-4, IL-10), alongside mitigating liver pathological damage. Additionally, it influenced the Nrf2/HO-1 pathway and reduced levels of apoptosis-related proteins. In vitro, the compounds delphinidin-3-O-glucoside (D3G) and cyanidin-3-O-glucoside (C3G) in BSSC were found to modulate cytokine levels, suggesting their role in ALI protection. The study concludes that BSSC extract, particularly due to D3G and C3G, effectively protects against LPS-induced ALI in mice by inhibiting inflammation, oxidative stress, and apoptosis.

Influence of preheating on antioxidant activity of the water extract from black soybean and color and sensory properties of black soybean decoction.

BACKGROUND: Our previous study indicated that black soybean (BS) tea (BST) made from preheated BS showed a significant protective effect against CCl4 -induced liver damage in Sprague Dawley rats due to BS's antioxidant activity. In this study, we investigated three different preheating methods on the antioxidant activity of water extract from BS (WBS) (Glycine max (L.) Merrill), as well as on the color and sensory properties of BST. RESULTS: WBS baked at 130 °C for 5 min showed high total phenol, flavonoid and anthocyanin contents among three preheating methods, as well as high DPPH radical-scavenging activity, ferrous ion chelating ability and reducing power. BST baked at 130 °C for 5 min had higher polyphenol oxidase activity and L*, a* and b* values but lower peroxidase activity then BST made from unheated BS. Sensory evaluation indicated that BST made from baked BS produced better aroma and flavor than the tea made from unheated BS or a commercial preheated BS. CONCLUSION: An appropriate preheating process improved the amount of extractable antioxidant and the antioxidant activity in WBS. BST baked at 130 °C for 5 min had good flavor properties, as well as high amount of antioxidant.