Effects of roasting and ultrasound-assisted enzymatic treatment of Nigella sativa L. seeds on thymoquinone in the oil and antioxidant activity of defatted seed meal.

BACKGROUND: Black cumin seeds (black seed; BS) contain various bioactive compounds, such as thymoquinone (TQ). Roasting and ultrasound-assisted enzymatic treatment (UAET) as pre-treatments can increase the phytochemical content in the BS oil. This study aimed to investigate the effects of pre-treatments on the TQ content and the yield of the BS oil and to profile the composition of defatted BS meal (DBSM), followed by evaluating antioxidant properties of the DBSM. RESULTS: The extraction yield of crude oil from BS was not affected by the roasting time. The highest extraction yield (47.8 ± 0.4%) was obtained with UAET cellulase-pH 5 (enzyme concentration of 100%). Roasting decreased the TQ content of the oil, while the UAET cellulase-pH 5 treatment with an enzyme concentration of 100% yielded the highest TQ (125.1 ± 2.7 µg mL-1 ). Additionally, the UAET cellulase-pH 5 treatment increased total phenolics and flavonoids of DBSM by approximately two-fold, compared to roasting or ultrasound treatment (UT) alone. Principal component analysis revealed that the UAET method might be more suitable for extracting BS oil with higher TQ content than roasting and UT. CONCLUSION: Compared to roasting or UT, using ultrasound along with cellulase could improve the oil yield and TQ in the oil from BS and obtain the DBSM with higher phenolics, flavonoids, and antioxidant activity. © 2023 Society of Chemical Industry.

Effects of black raspberry extract on gut microbiota, microbial metabolites, and expressions of the genes involved in cholesterol and bile acid metabolisms in rats fed excessive choline with a high-fat diet.

In our previous study, black raspberry (BR) reduced the serum levels of trimethylamine-N-oxide and cholesterol in rats fed excessive choline with a high-fat diet (HFC). We hypothesized that gut microbiota could play a crucial role in the production of trimethylamine and microbial metabolites, and BR could influence gut microbial composition. This study aimed to elucidate the role of BR on changes in gut microbiota and microbial metabolites in the rats. The phylogenetic diversity of gut microbiota was reduced in the rats fed HFC, while that in the BR-fed group was restored. The BR supplementation enriched Bifidobacterium and reduced Clostridium cluster XIVa. In the BR-fed group, most cecal bile acids and hippuric acid increased, while serum lithocholic acid was reduced. The BR supplementation upregulated Cyp7a1 and downregulated Srebf2. These results suggest that BR extract may change gut bacterial community, modulate bile acids, and regulate gene expression toward reducing cholesterol. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-023-01267-4.

Phenolics in buckwheat hull extracts and their antioxidant activities on bulk oil and emulsions.

Buckwheat hulls are discarded as waste, although they have more phenolic compounds than buckwheat groats. The antioxidant activities of buckwheat hull extracts prepared with water, 50% ethanol, and 100% ethanol were investigated in bulk oil, oil-in-water (O/W), and water-in-oil (W/O) emulsions. The relationship between the phenolic compositions of the extracts and their antioxidant activities in the three different lipid systems was also evaluated. Fifty percent ethanol extract had the highest total phenolic content (327 mg gallic acid equivalent [GAE]/g extract) followed by water and 100% ethanol extracts (211 and 163 mg GAE/g extract, respectively). The total oxidation rate (k) was not significantly different among the bulk oils added with the buckwheat hull extracts. However, in the O/W emulsion, the k was more reduced by the 50% and 100% ethanol extracts than by the water extract at the concentration of 100 µg GAE/g (2.9, 2.8, and 3.7 Totox/day, respectively). The k of the W/O emulsion was more reduced by the 100% ethanol extract than by the water and 50% ethanol extract at the concentration of 100 µg GAE/g (3.8, 4.7, and 4.5 Totox/day, respectively). Multivariate statistical analysis revealed that the contents of phenolic acids and their derivatives were the highest in the water extract among the extracts, while the contents of flavonoid glycosides and methylated polyphenols were the highest in the 50% and 100% ethanol extracts, respectively. The results suggest that flavonoid glycosides and methylated polyphenols could be potential candidates for retarding the oxidation of the emulsion system. PRACTICAL APPLICATION: Buckwheat hull extracts could retard lipid oxidation. Flavonoid glycosides and methylated polyphenols in buckwheat hull extracts may have an antioxidative effect on lipids. Thus, buckwheat hulls could be used as an antioxidant in lipid systems, as flavonoid glycosides and methylated polyphenols are properly extracted from buckwheat hulls.

Antibacterial Photodynamic Inactivation of Fagopyrin F from Tartary Buckwheat (Fagopyrum tataricum) Flower against Streptococcus mutans and Its Biofilm.

The objective of this study was to determine reactive oxygen species (ROS) produced by fagopyrin F-rich fraction (FFF) separated from Tartary buckwheat flower extract exposed to lights and to investigate its antibacterial photodynamic inactivation (PDI) against Streptococcus mutans and its biofilm. ROS producing mechanisms involving FFF with light exposure were determined using a spectrophotometer and a fluorometer. S. mutans and its biofilm inactivation after PDI treatment of FFF using blue light (BL; 450 nm) were determined by plate count method and crystal violet assay, respectively. The biofilm destruction by ROS produced from FFF after exposure to BL was visualized using confocal laser scanning microscopy (CLSM) and field emission scanning electron microscope (FE-SEM). BL among 3 light sources produced type 1 ROS the most when applying FFF as a photosensitizer. FFF exposed to BL (5 and 10 J/cm2) significantly more inhibited S. mutans viability and biofilm formation than FFF without the light exposure (p < 0.05). In the PDI of FFF exposed to BL (10 J/cm2), an apparent destruction of S. mutans and its biofilm were observed by the CLSM and FE-SEM. Antibacterial PDI effect of FFF was determined for the first time in this study.

Protective Effects of Black Raspberry (Rubus occidentalis) Extract against Hypercholesterolemia and Hepatic Inflammation in Rats Fed High-Fat and High-Choline Diets.

Choline is converted to trimethylamine by gut microbiota and further oxidized to trimethylamine-N-oxide (TMAO) by hepatic flavin monooxygenases. Positive correlation between TMAO and chronic diseases has been reported. Polyphenols in black raspberry (BR), especially anthocyanins, possess various biological activities. The objective of this study was to determine the effects of BR extract on the level of choline-derived metabolites, serum lipid profile, and inflammation markers in rats fed high-fat and high-choline diets. Forty female Sprague-Dawley (SD) rats were randomly divided into four groups and fed for 8 weeks as follows: CON (AIN-93G diet), HF (high-fat diet), HFC (HF + 1.5% choline water), and HFCB (HFC + 0.6% BR extract). Serum levels of TMAO, total cholesterol, and low-density lipoprotein (LDL)-cholesterol and cecal trimethylamine (TMA) level were significantly higher in the HFC than in the HFCB. BR extract decreased mRNA expression of pro-inflammatory genes including nuclear factor-κB (NF-κB), interleukin (IL)-1ß, IL-6, and cyclooxygenase-2 (COX-2), and protein expression of NF-κB and COX-2 in liver tissue. These results suggest that consistent intake of BR extract might alleviate hypercholesterolemia and hepatic inflammation induced by excessive choline with a high-fat diet via lowering elevated levels of cecal TMA and serum TMAO in rats.

Anti-Obesity Effect of Standardized Extract of Microalga Phaeodactylum tricornutum Containing Fucoxanthin.

Fucoxanthin (FX), a marine carotenoid found in macroalgae and microalgae, exhibits several beneficial effects to health. The anti-obesity activity of FX is well documented, but FX has not been mass-produced or applied extensively or commercially because of limited availability of raw materials and complex extraction techniques. In this study, we investigated the anti-obesity effect of standardized FX powder (Phaeodactylum extract (PE)) developed from microalga Phaeodactylum tricornutum as a commercial functional food. The effects of PE on adipogenesis inhibition in 3T3-L1 adipocytes and anti-obesity in high-fat diet (HFD)-fed C57BL/6J mice were evaluated. PE and FX dose-dependently decreased intracellular lipid contents in adipocytes without cytotoxicity. In HFD-fed obese mice, PE supplementation for six weeks decreased body weight, organ weight, and adipocyte size. In the serum parameter analysis, the PE-treated groups showed attenuation of lipid metabolism dysfunction and liver damage induced by HFD. In the liver, uncoupling protein-1 (UCP1) upregulation and peroxisome proliferator activated receptor γ (PPARγ) downregulation were detected in the PE-treated groups. Additionally, micro computed tomography revealed lower fat accumulation in PE-treated groups compared to that in the HFD group. These results indicate that PE exerts anti-obesity effects by inhibiting adipocytic lipogenesis, inducing fat mass reduction and decreasing intracellular lipid content, adipocyte size, and adipose weight.

Black Raspberry Seed Oil Improves Lipid Metabolism by Inhibiting Lipogenesis and Promoting Fatty-Acid Oxidation in High-Fat Diet-Induced Obese Mice and db/db Mice.

The objective of this study was to evaluate the beneficial effect of α-linolenic acid-rich black raspberry seed (BRS) oil on lipid metabolism in high-fat diet (HFD)-induced obese and db/db mice. Five-week-old C57BL/6 mice were fed diets consisting of 50% calories from lard, 5% from soybean, and 5% from corn oil (HFD), or 50% calories from lard and 10% from BRS oil (HFD + BRS oil diet) for 12 weeks. Six-week-old C57BL/KsJ-db/db mice were fed diets consisting of 16% calories from soybean oil (standard diet), 8% from soybean, and 8% from BRS oil, or 16% from BRS oil for 10 weeks. The BRS oil diets lowered the levels of triacylglycerol, nonesterified fatty acids, and total cholesterol in serum and liver of both of the obese and db/db mice as compared with the HFD and standard diet, respectively. mRNA levels of lipogenesis markers including cluster of differentiation 36, fatty-acid-binding protein 1, sterol regulatory element binding protein 1c, fatty-acid synthase, and solute carrier family 25 member 1 in the liver of the BRS oil groups were lower than those in the liver of the HFD and standard groups in the obese and db/db mice, respectively. On the other hand, fatty-acid oxidation markers including carnitine palmitoyltransferase 1A, acyl-CoA dehydrogenase, hydroxylacyl-CoA dehydrogenase α, and acyl-CoA oxidase in the liver of the BRS oil groups were higher than those in the liver of the HFD and standard groups in the obese and db/db mice, respectively. Peroxisome proliferator-activated receptor α mRNA and protein levels increased in the liver and epididymal adipose tissue of the obese and db/db mice fed BRS oil compared with HFD and standard diet, respectively. BRS oil might improve lipid metabolism by inhibiting lipogenesis and promoting fatty-acid oxidation in HFD-induced obese and db/db mice.

Adaptive neuro-fuzzy inference system-applied QSAR with bond dissociation energy for antioxidant activities of phenolic compounds.

The aim of this study was to develop quantitative structure-activity relationship (QSAR) models for predicting antioxidant activities of phenolic compounds. The bond dissociation energy of O-H bond (BDE) was calculated by semi-empirical quantum chemical methods. As a new parameter for QSAR models, sum of reciprocals of BDE of enol and phenol groups (X BDE ) was calculated. Significant correlations were observed between X BDE and antioxidant activities, and X BDE was introduced as a parameter for developing QSAR models. Linear regression-applied QSAR models and adaptive neuro-fuzzy inference system (ANFIS)-applied QSAR models were developed. QSAR models by both of linear regression and ANFIS achieved high prediction accuracies. Among the developed models, ANFIS-applied models achieved better prediction accuracies than linear regression-applied models. From these results, the proposed parameter of X BDE was confirmed as an appropriate variable for predicting and analysing antioxidant activities of phenolic compounds. Also, the ANFIS could be applied on QSAR models to improve prediction accuracy.

Dietary Black Raspberry Seed Oil Ameliorates Inflammatory Activities in db/db Mice.

The objective of this study was to evaluate the status of the markers related to inflammation in db/db mice fed black raspberry seed (BRS) oil, which is rich in α-linolenic acid. Mice were divided into four groups: (1) C57BL/6 mice fed 16 % calories from soybean oil (normal CON); (2) C57BL/KsJ-db/db mice fed 16 % calories from soybean oil (CON); (3) C57BL/KsJ-db/db mice fed 8 % calories from soybean and 8 % calories from BRS oil (BRS 50 %); and (4) C57BL/KsJ-db/db mice fed 16 % calories from BRS oil (BRS 100 %). After 10 weeks, n-6/n-3 fatty acid ratios were significantly (P < 0.05) lower in the livers and epididymal adipose tissues of the BRS 50 % and BRS 100 % mice than in the CON. Serum TNFα and IL-6 were significantly (P < 0.05) lower in the BRS 50 % and BRS 100 % than in the CON. Serum IL-10 was significantly (P < 0.05) higher in the BRS 100 % than the CON. In the liver and epididymal adipose tissue, mRNA levels of pro-inflammatory markers in the BRS 50 % and BRS 100 % were lower than in the CON. Anti-inflammatory markers were higher in the epididymal adipose tissues of the BRS 50 % and BRS 100 % than in the CON. In the epididymal adipose tissue, macrophage infiltration markers (F4/80 and CD68) and leptin mRNA were significantly (P < 0.05) lower in the BRS 50 % and BRS 100 % than in the CON. Results of this study suggest that BRS oil may have anti-inflammatory effects in obese diabetic mice by ameliorating inflammatory responses.

Chemopreventive activity of ellagitannins and their derivatives from black raspberry seeds on HT-29 colon cancer cells.

Black raspberry (BRB) seeds are a major waste product after fruit processing. The seeds are abundant in ellagitannins (ET), a class of hydrolysable tannins, which are hydrolyzed to ellagic acid (EA) and further metabolized to urolithin A (UA) and urolithin B (UB), known to be bioavailable in the colon and the prostate. In this study, the anti-cancer activities of these compounds were evaluated on HT-29 colon cancer cells. ET, EA, UA and UB inhibited the proliferation of the cancer cells. EA caused a slight, but significant cell cycle arrest at the G1 phase, and urolithins caused cell cycle arrest at the G2/M phase and upregulated p21 expression. Apoptotic cells were detected by Annexin V-FITC/PI assay when treated with the compounds. Disruption in mitochondrial membrane potential and activation of caspases 8 and 9 suggest that both extrinsic and intrinsic apoptotic pathways may be involved. Activation of caspase 3 and cleavage of PARP further confirmed the induction of the apoptosis. ET, EA, UA and UB showed anti-cancer activity by arresting the cell cycle and inducing apoptosis on HT-29 human colon cancer cells. This study suggests that the BRB seeds could be a potential source of anti-cancer ET.

The anti-obesity effects of the dietary combination of fermented red ginseng with levan in high fat diet mouse model.

In this study, to evaluate the anti-obesity effects of fermented red ginseng (FG), levan (L), and their combination (FGL), we investigated their effects on the weights of body, liver and white adipose tissue, lipid profiles, and biomarkers for insulin resistance in high fat diet (HFD)-induced obese C57BL/6J male mice. Furthermore, the levels of leptin in the serum were measured. FG (150 mg/kg/d), L (100 mg/kg/d), and FGL (150 mg/kg/d of FG plus 100 mg/kg/d of L) were administered orally to mice daily for 11 weeks. After 11 weeks feeding, FGL showed significantly lower body weight and fat mass with decreasing food efficiency ratio than the HFD control mice. In addition, the FGL group was significantly lower in the levels of total cholesterol and fasting blood glucose and score of the homeostatic model assessment of insulin resistance. Furthermore, FGL decreased serum leptin levels compared to the HFD control group. Taken together, FGL showed a significant anti-obesity effect in HFD-induced obese mice and prevent insulin and leptin resistance. FGL may be potentially useful for the prevention of obesity.

Unrefined and refined black raspberry seed oils significantly lower triglycerides and moderately affect cholesterol metabolism in male Syrian hamsters.

Unrefined and refined black raspberry seed oils (RSOs) were examined for their lipid-modulating effects in male Syrian hamsters fed high-cholesterol (0.12% g/g), high-fat (9% g/g) diets. Hamsters fed the refined and the unrefined RSO diets had equivalently lower plasma total cholesterol and high-density lipoprotein (HDL) cholesterol in comparison with the atherogenic coconut oil diet. The unrefined RSO treatment group did not differ in liver total and esterified cholesterol from the coconut oil-fed control animals, but the refined RSO resulted in significantly elevated liver total and esterified cholesterol concentrations. The unrefined RSO diets significantly lowered plasma triglycerides (46%; P=.0126) in comparison with the coconut oil diet, whereas the refined RSO only tended to lower plasma triglyceride (29%; P=.1630). Liver triglyceride concentrations were lower in the unrefined (46%; P=.0002) and refined (36%; P=.0005) RSO-fed animals than the coconut oil group, with the unrefined RSO diet eliciting a lower concentration than the soybean oil diet. Both RSOs demonstrated a null or moderate effect on cholesterol metabolism despite enrichment in linoleic acid, significantly lowering HDL cholesterol but not non-HDL cholesterol. Dramatically, both RSOs significantly reduced hypertriglyceridemia, most likely due to enrichment in α-linolenic acid. As a terrestrial source of α-linolenic acid, black RSOs, both refined and unrefined, provide a promising alternative to fish oil supplementation in management of hypertriglyceridemia, as demonstrated in hamsters fed high levels of dietary triglyceride and cholesterol.

Antiproliferation properties of grain sorghum dry distiller's grain lipids in Caco-2 cells.

Antiproliferative properties of lipids extracted from grain sorghum (GS) dry distiller's grain (DDG) were analyzed to determine the feasibility of developing GS coproducts as a source for human health dietary ingredients. The lipid extract of GS-DDG was delivered to human colon carcinoma (Caco-2) cells by solubilizing 0-1000 microg/mL of GS-DDG lipids in 100 microg/mL increments with micelles. A significant reduction in cell viability (25-50%) resulted at treatment levels of 400-1000 microg/mL GS-DDG lipids (p < 0.05). Alternatively, total protein levels of cells treated with 400, 500, and 600 microg/mL of GS-DDG lipid were not significantly different from the control, indicating cell growth during the treatment period. Total cell counts for the control were not significantly different from the GS-DDG lipid treated cells, but dead cell counts increased by approximately 10% for the latter sample with a concomitant increase of the intercellular protein lactate dehydrogenase leakage (30-40%) in the medium. Preliminary analysis by the fluorescence-activated cell method (FACs) demonstrated that nonviable cells were in either the early apoptotic, late apoptotic, or necrotic stage post-treatment with 400, 500, and 600 microg/mL GS-DDG lipids. Physiochemical characterization of the GS-DDG lipids used for the antiproliferation study showed the presence of vitamin E (predominantly gamma-tocopherol), triacylglycerides (predominantly linoleic acid), policosanols, aldehydes, and sterols (predominantly campesterol and stigmasterol), each of which or as synergistic/additive group of constituents may be responsible for the antiproliferative effect.