Identification of natural, bioactive compounds has been of interest to the biomedical field, in particular for the treatment of cancer. Flavonoids are a family of compounds, present across many plant species, known for their polyphenolic structure that results in cellular bioactivity. The flavonoid, sakuranetin, is produced due to external stressors, through the naringenin biosynthetic pathway. Binding affinity and docking simulations confirmed sakuranetin binding to the estrogen receptor alpha (ERα) pocket in a manner similar to, but with weaker affinity than 17β-estradiol (E2). Due to sakuranetin having chemical similarities to estrogen, this study evaluated the potential of sakuranetin to act as an endocrine modulator in estrogen receptor-positive (ERα+) breast cancer cell lines. In the ERα + cell lines, MCF-7 and T-47D, sakuranetin treatment induced dose-dependent estrogenic activity (≥ 10 μM). Sakuranetin significantly increased colony formation and cell proliferation in the MCF-7 cell line. Breast cancer cell lines with constitutively active ER through an inserted mutation in the ERα (Y537S) demonstrated no significant changes in estrogenic activity or cellular proliferation following sakuranetin treatment, except at an elevated dose (50 μM). Finaly, sakuranetin treatment enhanced genes associated with ER signaling and significantly increased ERα-mediated gene (PGR and CXCL12) expression. These results support the role of sakuranetin as a natural estrogenic compound.

Kudzu's invasive nature has contributed to its classification as a weed, as it frequently outcompetes native plant species, leading to extensive overgrowth. Efforts to control kudzu have proven challenging, with moderate success using physical or biological agents. In this study, we evaluated the effects of two such control agents, ultraviolet C radiation and Myrothecium verrucaria, to significantly increase the production of tuberosin, a phytoalexin isoflavone. Our findings demonstrate that estrogenic activity of tuberosin is cell-type-dependent, displaying antagonist or competitive inhibition when combined with 17-β-estradiol in the estrogen receptor (ER) positive cell lines MCF-7 and T-47D, while showing dose-dependent agonist activity in HEK293 cells transfected to express both ER receptors (α and β). Tuberosin was shown to modulate ER pathways, alter ER-mediated gene expression, and increase cell proliferation in a dose-dependent manner while maintaining expression of the ERα protein. Binding affinity and docking simulations confirmed tuberosin binding to the ERα pocket in a similar but weaker manner compared to synthetic estrogen. Tuberosin-treated endothelial cells suppressed vascular network assembly and maturation without affecting the cellular proliferative capacity. The presented studies leverage current kudzu management methods to naturally produce tuberosin, examine cell-type-specific effects, and support further investigation as an antiestrogen for breast cancer treatment.

Naringenin (Nar) is a citrus fruit-derived phytoestrogen, a group of dietary compounds produced by a wide variety of plants. Due to structural similarity to 17‐β‐estradiol (E2), phytoestrogens can bind to estrogen receptors (ERs) to exert context-dependent estrogenic and/or anti-estrogenic effects. As such, there are potential health benefits and risks associated with phytoestrogen exposure. To investigate the effects of Nar in breast cancer, based on its activity as a phytoestrogen, we treated MCF-7 ER-positive (ER+) breast cancer cells with Nar (10 µM) and examined transcriptomic changes in MCF-7 cells induced by Nar treatment. Pathway analysis included in our dataset shows upregulation of genes associated with estrogen signaling and epithelial to mesenchymal transition in breast cancer cells treated with naringenin.

The nutritional benefits of nut consumption are complicated by the presence of allergens. Food processing techniques can modify the food protein properties by altering their biophysical characteristics and allergen activity. This study examines the phenylglyoxal-based chemical modification of the cashew nut allergen Ana o 3. Immunoassays with multiple antibodies demonstrate reduced recognition of phenylglyoxal-modified Ana o 3. Mass spectrometry identified multiple Ana o 3 modification sites, including arginine 41, 54, 85, and 111. Circular dichroism, biochemical assays, and molecular simulation indicate that the modifications resulted in minimal protein structure alteration. The research presented here provides insight into Ana o 3 surface chemistry and structure, and it could be applied in the design of alternative forms of immunotherapy to treat cashew nuts and other food allergies.

Along with dairy allergic/intolerant and vegetarian/vegan individuals, flexitarian consumers contribute to the market for plant-based milk alternatives. Niche pigmented (purple and red) and/or aromatic (aromatic brown and white) rice varieties were used to formulate ricemilks, which were compared to a product made from commercial nonaromatic white rice. Descriptive sensory analysis (9 trained panelists) and affective sensory testing (161 consumers) were used to explore perceptions of ricemilks. Additional sample differences were characterized by instrumental color, viscosity, 2-acetyl-1-pyrroline (2-AP), starch quality, total phenolic content (TPC), oxygen radical absorbance capacity (ORAC), and proximate analyses. Compared to nonaromatic white ricemilk, increased bioactive and antioxidant potential was exhibited for pigmented varieties (TPC ranging 56.6-64.5 mu g/mL; ORAC ranging 628-753 mu M Trolox equivalents). However, the pigments also resulted in decreased acceptability of product color. For red ricemilk, this was overcome after tasting, resulting in the highest grouping of overall liking scores along with aromatic brown and aromatic white ricemilks (5.6-5.7 on a 9-point hedonic scale)-all associated with a desirable "popcorn" aromatic from 2-AP. Highest purchase intent was found for aromatic brown ricemilk (47% overall and 69% among regular dairy and nondairy milk consumers) which was associated with the "dairy" aromatic. Consumers also favored colors and mouthfeel more reminiscent of dairy milk. Ricemilks made from niche pigmented and/or aromatic rices provided unique flavors and outperformed nonaromatic white ricemilk in overall acceptability. Ricemilks made from aromatic whole grain varieties can offer a new health-promoting functional beverage option to consumers, and a new use of niche rice varieties.

Background and ObjectivesRice bran is a source of bioactive polyphenols. This study aimed to characterize the antidiabetic potential of different rice brans (one brown, two red, and two purple) by examining their ability to inhibit alpha-amylase, alpha-glucosidase, and pancreatic lipase, and to stimulate glucose uptake in adipocytes.FindingsAll pigmented bran extracts significantly inhibited alpha-glucosidase. Both red rice brans inhibited alpha-amylase close to 50% at the highest dose tested. All four pigmented rice brans inhibited lipase in vitro (IC50 values 4.38-10.09 mg/mL). The pigmented rice brans studied contained higher levels of total polyphenolic content (TPC), total flavonoid content (TFC), and antioxidant capacity when compared to brown rice bran. Purple bran contained anthocyanins and red brans contained proanthocyanidins. The pigmented brans consisted of higher levels of total benzoic acids, while the brown rice bran had higher levels of total cinnamic acids. Tricin was identified in all pigmented brans.ConclusionsOur study indicates that pigmented rice brans have the ability to manage obesity and diabetes due to their inhibitory effects on lipid and carbohydrate digestion. Several bioactive polyphenols were identified.Significance and NoveltyThe potential of pigmented rice bran to inhibit alpha-amylase, alpha-glucosidase, and lipase was confirmed.

Xenohormesis proposes that phytochemicals produced to combat stressors in the host plant exert biochemical effects in animal cells lacking cognate receptors. Xenohormetic phytochemicals such as flavonoids and phytoalexins modulate a range of human cell signaling mechanisms but functional correlations with human pathophysiology are lacking. Here, potent inhibitory effects of grapefruit-derived Naringenin (Nar) and soybean-derived Glyceollins (Gly) in human microphysiological models of bulk tissue vasculogenesis and tumor angiogenesis are reported. Despite this interference of vascular morphogenesis, Nar and Gly are not cytotoxic to endothelial cells and do not prevent cell cycle entry. The anti-vasculogenic effects of Glyceollin are significantly more potent in sex-matched female (XX) models. Nar and Gly do not decrease viability or expression of proangiogenic genes in triple negative breast cancer (TNBC) cell spheroids, suggesting that inhibition of sprouting angiogenesis by Nar and Gly in a MPS model of the (TNBC) microenvironment are mediated via direct effects in endothelial cells. The study supports further research of Naringenin and Glyceollin as health-promoting agents with special attention to mechanisms of action in vascular endothelial cells and the role of biological sex, which can improve the understanding of dietary nutrition and the pharmacology of phytochemical preparations.

Legumes are a rich source of polyphenolic compounds known for their ability to promote health. Under stress conditions, legumes have been shown to produce higher levels of secondary metabolites, as a defensive mechanism. Hence, the present study aimed to induce legume seeds (e.g., soybean, chickpea, green pea, and red kidney bean) by inoculating them with Aspergillus sojae (A. sojae) and to evaluate the extracts for phytoalexins, phenolics, and antioxidant, antiobesity, and antidiabetic potentials. The UPLC-DAD findings of A. sojae-induced legumes showed medicarpin and maackiain in chickpea, pisatin in green pea, glyceollin I-III in soybean, and kievitone and phaseollin in red kidney bean. All induced legumes exhibited a higher total polyphenol content compared to the non-induced ones. Among induced legumes, soybean exhibited a higher (4.85 mg GAE/g) polyphenol content. The UPLC-ESI-QTOF-MS/MS findings established that legumes contained substantial levels of protocatechuic acid, vanillic acid, ferulic acid, chlorogenic acid, coumaric acid, 4-hydroxybenzoic acid, and caffeic acid. The results of antioxidant assays revealed a significantly higher level of activity in induced red kidney bean and soybean, whereas the level of activity in non-induced legumes was markedly lower. Moreover, induced red kidney bean effectively inhibited α-glucosidase (87.2%) and α-amylase (63.90%) at 5 mg/mL. Additionally, the maximum lipase inhibitory effects were displayed by induced soybean (72.54%) at 20 mg/mL.

Legumes are a predominant source of isoflavones, termed phytoestrogens, that mimic 17 beta-estradiol (E2). Phytoalexins are inducible isoflavones produced in plants subjected to environmental stressors (e.g., UV, heat, or fungi). This study investigated estrogenic activity of snow and green peas elicited with Aspergillus sojae. Elicited extracts increased estrogenic activity and proliferation of breast cancer cells (MCF-7 or T47D) in a dose-dependent manner but exhibited antiestrogenic activity when combined with synthetic E2. HPLC analysis of elicited pea extracts identified (+)-pisatin as the primary phytoalexin, which was produced significantly (p < 0.0001) more in snow pea compared to green pea. RNA sequencing results suggested potential functional effects on endothelial cells and tissue vascularization. Indeed, (+)-pisatin enhanced metrics of network assembly and maturation in a microphysiological model of bulk tissue vasculogenesis. Thus, context-dependent functional effects of (+)-pisatin and pharmacologically similar phytoestrogens on the entire tissue microenvironment should be considered in preclinical investigation as potential therapeutic agents.

Glyceollins are soy-derived phytoalexins that have been proposed as candidate compounds for the prevention of prostate cancer. The present study tested the efficacies of glyceollins on prostate cancer prevention in-vitro and in-vivo. In-vitro, glyceollins significantly inhibited the androgen-responsive LNCaP cell growth consistent with inhibition of the androgen-mediated pathway. In-vivo, dietary glyceollins attenuated LNCaP tumor xenograft growth in a nude mouse model and correlated with the inhibition of tumor cells proliferative marker PCNA mRNA levels. However, unlike in-vitro, dietary glyceollins did not affect marker genes for the androgen- responsive pathway, cell cycle, and angiogenesis in the tumor xenograft. Dietary glyceollins also did not affect the marker genes for xenobiotic metabolism, cholesterol transport, or inflammatory pathways in liver. The low bioavailability of glyceollins (0.054 +/- 0.013 mu M in the plasma) might have led to the lack of effectiveness of glyceollins on the marker genes in-vivo. . Interestingly, dietary glyceollins significantly lower the abundance of cecal Bifidobacterium, , butyrate producing bacteria, compared to the control diet. Thus, glyceollins act differently in-vitro and in-vivo. . The protective effects of glyceollins in-vivo may be independent of the androgen responsive pathway but related to modulation of the butyrate, a putative prostate cancer promoting agent, production by the gut microbiome.

As breast cancer cells transition from letrozole-sensitive to letrozole-resistant, they over-express epidermal growth factor receptor (EGFR), mitogen-activated protein kinase (MAPK), and human epidermal growth factor receptor 2 (HER2) while acquiring enhanced motility and epithelial-to-mesenchymal transition (EMT)-like characteristics that are attenuated and reversed by glyceollin treatment, respectively. Interestingly, glyceollin inhibits the proliferation and tumor progression of triple-negative breast cancer (TNBC) and estrogen-independent breast cancer cells; however, it is unlikely that a single phytochemical would effectively target aromatase-inhibitor (AI)-resistant metastatic breast cancer in the clinical setting. Since our previous report indicated that the combination of lapatinib and glyceollin induced apoptosis in hormone-dependent AI-resistant breast cancer cells, we hypothesized that combination therapy would also be beneficial for hormone independent letrozole-resistant breast cancer cells (LTLT-Ca) compared to AI-sensitive breast cancer cells (AC-1) by decreasing the expression of proteins associated with proliferation and cell cycle progression. While glyceollin + lapatinib treatment caused comparable inhibitory effects on the proliferation and migration in both cell lines, combination treatment selectively induced S and G2/M phase cell cycle arrest of the LTLT-Ca cells, which was mediated by decreased cyclin B1. This phenomenon may represent a unique opportunity to design novel combinatorial therapeutic approaches to target hormone-refractory breast tumors.

Sumac sorghum (Sorghum bicolor L. Moench) bran contains polyphenolic compounds with health-promoting properties. Functional food product development can add economic value to sorghum bran, but acceptability may be limited by bitter taste. An acidified cold-brewed sorghum bran beverage was developed, and simplified sweetened (SB) and unsweetened (UB) versions were analyzed for antioxidant profiles (total phenolic content, phenolic profiling via HPLC, condensed tannins, oxygen radical absorbance capacity, and 2,2-diphenyl-1-picrylhydrazyl radical scavenging) and sensory properties. Beverages contained condensed tannins (average of 0.33 mg catechin equivalents per milliliter) along with other flavonoids and demonstrated in vitro antioxidant capacity (3.32-3.96 mmol Trolox equivalents per milliliter). One hundred fourteen consumers, grouped by 6-n-propylthiouracil (bitterness) taster status, rated acceptability (9-point hedonic scales), bitterness intensity (5-point scale), and purchase intent (PI; yes/no) of model beverages. Although the concept was novel, SB was clearly more acceptable to consumers regardless of preference for sweet versus unsweet tea (as a reference product concept). Mean mouthfeel and flavor liking scores were significantly higher for SB (6.2 and 5.1, respectively) than UB (4.8 and 3.0, respectively). Bitterness was suppressed by the sweetness in SB (mean bitterness intensity of 1.8 vs. 2.6 for UB), although 21% of consumers still found it "too bitter." However, consumers' taster status did not affect beverage acceptability nor bitterness perceptions at present phenolic levels (0.53 gallic acid equivalents per milligram). After a sorghum/antioxidant information message, positive PI increased to 56% for SB, impacting more females than males. Enhanced familiarity and effective product messaging can improve acceptability of acidified cold-brewed sorghum bran beverages as a novel functional food product concept. Practical ApplicationNew food applications of cereal bran can provide value in terms of byproduct utilization and delivery of health-promoting ingredients. Sorghum bran beverages demonstrated antioxidant activity, and the sweetened formulation showed decreased bitterness and increased acceptability. Consumers were positively influenced by antioxidant information, although most had not previously consumed sorghum.

Soybean products have nutrients, dietary fiber, and phytoalexins beneficial for cardiovascular and overall health. Despite their high consumption in Asian populations, their safety in Western diets is debated. We conducted a dose-escalating clinical trial of the safety and tolerability of soybean products in eight older adults (70–85 years) with obesity. Whole green soybean pods grown under controlled conditions were processed to flour (WGS) at the United States Department of Agriculture using common cooking techniques such as slicing and heat treatment. WGS incorporated into food products was consumed at 10 g, 20 g, and 30 g/day for one week at each dose. The gastrointestinal outcomes, clinical biomarkers, and adverse events were evaluated. We explored the stimulation of phytoalexin (glyceollin) production in live viable soybean seeds (LSS-G). We compared the compositions of WGS and LSS-G with commercial soybean flour and its fermented and enzymatically hydrolyzed forms. We found that although 30 g WSG was well-tolerated, and it made participants feel full. Our processing produced glyceollins (267 µg/g) in LSS-G. Processing soybean flour decreased the iron content, but reduced the oligosaccharides, which could attenuate flatulence. Providing soybean flour at <30 g/day may be prudent for overall health and to prevent the exclusion of other food groups and nutrients in older adults with obesity.

Over the last 30 years, thousands of articles have appeared examining the effects of soaking and germinating brown rice (BR). Variable germination conditions and methods have been employed to measure different health-beneficial parameters in a diverse germplasm of BR. Research results may therefore appear inconsistent with occasional anomalies, and it may be difficult to reach consensus concerning expected trends. Herein, we amassed a comprehensive review on germinated brown rice (GBR), attempting to codify 133 peer-reviewed articles regarding the effects on 164 chemical parameters related to health and nutrition in BR and in value-added food products. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA-2020) approach was used to direct the flow of the literature search. A pair-wise comparison t-test was performed to deliver an overall approach indicating when a given compound has been found to significantly increase or decrease through germination, which was grouped into GABA and polyamines, gamma-Oryzanol and phytosterols, phenolic compounds, vitamins, proteins and amino acids, starchy carbohydrates, free sugars, lipids, minerals and phytic acid. This resource will stimulate interest in germinating rice and optimistically help increase both production and consumption of highly nutritious, health-beneficial rice with pigmented bran.

Background and objectives Resistant starch (RS) has health-promoting properties, but can be altered during frying. The aim of this study was to examine the textural and sensory characteristics of fried rice batters with varying amounts of RS. Findings Adding resistant starch (RS2 and RS3) to rice batters or using a high RS rice flour significantly increased resistant starch contents after frying. The instrumental textural analysis of the fried batters indicated all of the fried rice flour batters used had higher hardness values and quantity of fractures when compared to wheat batter. Sensory evaluation using battered onion strips indicated that adding RS to rice flour batters did not significantly alter the attributes for hardness, fracturability, crispness, and toothpacking. However, the sensory evaluation for battered onion strips indicated the low RS rice flour displayed significantly higher values for hardness, fracturability, and crispness when compared to the high RS rice flour. Conclusions The addition of RS or the use of high RS rice flour in fried battered onion strips did not significantly alter the textural and sensory attributes that consumers enjoy. Significance and novelty Rice is a source of RS and can be included in fried food batters to promote better health.

Aromatase inhibitors (AIs) are standard treatment for estrogen-dependent postmenopausal breast tumors; however, resistance develops leading to tumor relapse and metastasis. We previously demonstrated that glyceollin inhibits proliferation, survival, and migration of hormone-independent letrozole-resistant breast cancer. Since many AI-resistant tumors remain hormone-dependent, identifying distinctions between estrogen-receptor-positive (ER+) and ER-negative (ER-) AI-resistant tumor response to therapy is critical. We hypothesize that treating ER+ letrozole-resistant T47D breast cancer cells (T47DaromLR) with a combination of 10 μM glyceollin and 0.5 μM lapatinib (a dual EGFR/HER2 inhibitor) will decrease cell proliferation through induction of apoptosis. The T47DaromLR cells were found to overexpress HER2 and MAPK while maintaining aromatase and ER levels compared to their letrozole-sensitive (T47Darom) counterparts. In the absence of estrogen stimulation, glyceollin ± lapatinib had no effect on the proliferation of the T47Darom cells, while glyceollin treatment caused 46% reduction in the proliferation of T47DaromLR cells, which was further diminished when combined with lapatinib. While neither agent influenced cell migration, glyceollin and lapatinib reduced S and G2/M phase cell entry and exclusively induced apoptosis by 1.29-fold in the T47DaromLR cells. Taken together, these results suggest that glyceollins and lapatinib may have potential as a novel combination therapeutic approach for hormone-dependent, letrozole-resistant tumors.

Environmentally sound plant treatments that can impose mild physiological stress and elicit bioaccumulation of useful phytochemicals such as kaempferols are limited. We tested ABA foliar application, 100 or 200 µM, and two types of leaf wounding, piercing or hole punching in young greenhouse-grown soy plants. Leaf gas exchange and A/Ci response, \(\phi\)PSII, pigments and antiradical activity were measured using the same leaf and kaempferols were measured in the leaf above. ABA 200 µM-treated plants had \(\geq\)20% less gas exchange and 17% less ETR, but greater Vcmax and Jmax compared with control. They had 55% and 100% more stomatal limitation to Pnet and \(\phi\)PSII, respectively, than the control. Leaf-wounded plants showed the lowest stomatal limitation to either Pnet or \(\phi\)PSII. Leaf piercing increased chlorophylls 39% and carotenoids 38% compared with the control. Six kaempferols quantified were found to be mono-, di- and triglycosides. Each leaf treatment increased total kaempferol yield ranging from 42% in ABA 100 µM to 68% in ABA 200 µM treatment compared with control. In general, kaempferol yields were positively correlated to Pnet in ABA 100 µM-treated plants and to gs in ABA 200 µM-treated plants but negatively correlated to Pnet in leaf-pierced plants. Results of this study show that appreciable increase in kaempferol yield and thus phytochemical or nutritional quality of soy leaves is achievable with environmentally friendly foliar treatments. ABA application and wounding affected the association between photosynthetic primary metabolism and kaempferol accumulation differently. Both ABA application and wounding are promising leaf treatments for eliciting kaempferol accumulation in young soy leaves.

Sweet sorghum syrup is non-GMO, gluten-free, and can be produced free of additives and preservatives. Knowledge of the nutritional and dietary content of sweet sorghum syrups compared to other common food-grade syrups will expand their utilization as a natural and nutritional food ingredient. Proximate food attributes and critical minerals were measured in commercial high fructose corn syrup (HFCS), corn, honey, maple, sugarcane, agave, rice, and white grain (non-tannin) sorghum syrups and compared to ten commercial sweet sorghum syrups. Agave syrup was the highest (P < 0.05) source of fat (mean 2.1%) although fat varied (P < 0.05) with the agave brand. Protein content of sweet sorghum syrups ranged from 0.9 to 4.0%, with the mean (1.80%) nearly two-folds higher than the other syrups (0.96%). Sweet sorghum syrups contained markedly higher (P < 0.05) mean amounts of magnesium (120 mg) than the other syrups (5 mg). Potassium contents of all the sweet sorghum syrups were also very high (up to 1710 mg) and markedly higher (P < 0.05) than all the other syrups. Iron was negligible in servings of the syrups except for sweet sorghum which contained over half (52.7%) of the daily recommended value (13 mg). In this limited study, sweet sorghum syrups were rich dietary sources of magnesium, potassium, calcium, iron, and contained negligible sodium.

The aim of this study was to evaluate the effects of resistant starch (RS) and fat levels on the gut microbiome in C57BL/6 mice. Three levels of RS from three varieties of rice were the major source of carbohydrates and fat levels were low (10%) and high (39%). We confirmed that RS decreased the Firmicutes to Bacteroidetes ratio, increased SCFA production by higher Bacteroidaceae and S24-7 abundance, and enriched predicted gene families of glycosidases and functional pathways associated with carbohydrate and glycan metabolism. We also found correlations between microbial taxa and tissue gene expression related to carbohydrate and lipid metabolism. Moreover, increasing RS levels resulted in a molecular ecological network with enhanced modularity and interspecific synergy, which is less sensitive to high fat intervention. Overall, RS as low as 0.44% from cooked rice can modulate gut microbiome in mice, which correlated to a protective effect against deleterious effects of an obesogenic diet.

Plant-based beverage consumption is increasing markedly. Value-added dehulled rice (Oryza sativa) germination was investigated to improve beverage qualities. Germinating brown rice has been shown to increase health-promoting compounds. Utilizing green processing, wholesome constituents, including bran, vitamins, minerals, oils, fiber and proteins should should convey forward into germinated brown rice beverages. Rapid visco-analyzer (RVA) data and trends established that brown rice, preheated brown rice and germinated brown rice had higher pasting temperatures than white rice. As pasting temperature in similar samples may be related to gelatinization, RVA helped guide the free-flowing processing protocol using temperatures slightly above those previously reported for Rondo gelatinization. Particle size analysis and viscometric evaluations indicate that the developed sprouted brown rice beverage is on track to have properties close to commercial samples, even though the sprouted brown rice beverage developed has no additives, fortifications, added oils or salts. Phenolics and γ-aminobutyric acid increased slightly in germinated brown rice, however, increases were not maintained throughout most stages of processing. Significantly lower inorganic arsenic levels (113 ng/g) were found in germinated (sprouted) brown rice, compared to Rondo white and brown rice, which is far below the USA threshold level of 200 ng/g.