Dietary supplementation of black rice bran to colon carcinogen-induced mice: Examining the development of colorectal cancer by improving environmental colon conditions.

This research aims to identify the effects of the administration of a black rice bran diet on colorectal cancer in dextran sodium sulfate and azoxymethane-induced BALB/c mice. The research was conducted on three groups consisting of eight Balb/c mice: two groups were fed with carcinogens, and the third group, referred to as the normal group, was supplied with Isotonic NaCl 0.9% intraperitoneally. One group fed with carcinogens was supplied a standard AIN 1993 M diet modified with black rice bran as a substitute of fibre source, while the other two mice groups were fed the standard diet (AIN-93M) containing cellulose fibre. At the 17th week, all mice were euthanized; their colonic sections were taken for histopathological evaluation, and cecum for short-chain fatty acids concentration, total lactic acid bacteria, pH and ß-glucuronidase activity evaluations. The results show an increase in the total lactic acid bacteria and short-chain fatty acids in the mice group fed with rice bran. Consequently, pH value and ß-glucuronidase activity had decreased. Histopathological evaluation of mucosal tissue exhibited inhibition of the tumor growth rate in the mice groups fed rice bran compared to the group supplied with the standard diet. Furthermore, the proliferating cell nuclear antigen expression had decreased significantly, while expression of caspase-8 and caspase-3 had increased notably, in the group fed with a rice bran diet. These results suggest that black rice bran can effectively inhibit colon carcinogenesis. The potential of black rice bran as a source of fibre has not been studied in detail regarding the inhibition mechanism of colorectal cancer cells; further investigation in this field could provide valuable information about new strategies to prevent colorectal cancer. This strand of research is very important to developing preventive methods against cancer and promoting the concept of healthy products, including functional foods.

Continuous production of velvet bean-based bioactive peptides in membrane reactor with dual enzyme system.

One of the main challenges hindering the commercialization of bioactive peptides is the lack of scalable and consistent production methods. To overcome this obstacle, an automated enzyme membrane reactor was used to continuously produce bioactive peptides from velvet bean (Mucuna pruriens). The optimum operating conditions were [E]/[S] = 5%, pH = 7.5, and τ = 12 h. The long-term continuous operation of the EMR system demonstrated its ability to maintain steady-state conditions. To minimize membrane fouling, an industrially viable strategy was employed, which combines operation at threshold flux and performing regular membrane cleaning. Further fractionation of the hydrolysates with a 2-kDa PES membrane resulted in the highest bioactivity. The IC50 values for antioxidant and ACE inhibition were 17.85 and 4.58 µg protein/mL, respectively. To map the overall bioactivities of the hydrolysates, LC-MS analysis coupled with BIOPEP-UWM database was performed and obtained DPP-4 and ACE inhibitors as the predominant bioactive activities.

Tryptamine, a Microbial Metabolite in Fermented Rice Bran Suppressed Lipopolysaccharide-Induced Inflammation in a Murine Macrophage Model.

Fermentation is thought to alter the composition and bioavailability of bioactive compounds in rice bran. However, how this process affects the anti-inflammatory effects of rice bran and the bioactive compounds that might participate in this function is yet to be elucidated. This study aimed to isolate bioactive compounds in fermented rice bran that play a key role in its anti-inflammatory function. The fermented rice bran was fractionated using a succession of solvent and solid-phase extractions. The fermented rice bran fractions were then applied to lipopolysaccharide (LPS)-activated murine macrophages to evaluate their anti-inflammatory activity. The hot water fractions (FRBA), 50% ethanol fractions (FRBB), and n-hexane fractions (FRBC) were all shown to be able to suppress the pro-inflammatory cytokine expression from LPS-stimulated RAW 264.7 cells. Subsequent fractions from the hot water fraction (FRBF and FRBE) were also able to reduce the inflammatory response of these cells to LPS. Further investigation revealed that tryptamine, a bacterial metabolite of tryptophan, was abundantly present in these extracts. These results indicate that tryptamine may play an important role in the anti-inflammatory effects of fermented rice bran. Furthermore, the anti-inflammatory effects of FRBE and tryptamine may depend on the activity of the aryl hydrocarbon receptor.

Increased γ-Aminobutyric Acid Content of Germinated Brown Rice Produced in Membrane Reactor.

RESEARCH BACKGROUND: Rice germination is a natural approach to enhance the physical and functional properties of brown rice. Thus, the aim of this study is to investigate the influence of different germination methods on functional properties of germinated brown rice and evaluate the process feasibility. EXPERIMENTAL APPROACH: Brown rice of IPB 3S variety was germinated with three different methods: (i) complete soaking without water replacement, (ii) complete soaking with water replacement every six hours, and (iii) complete soaking with continuous washing in the developed membrane-facilitated soaking reactor. RESULTS AND CONCLUSIONS: The application of the membrane reactor for producing germinated brown rice maintained the pH of the soaking solution relatively constant (i.e. 6.8-7.0). This indicated the circumvention of natural fermentation during brown rice germination. Moreover, the mass fraction of γ-aminobutyric acid in germinated brown rice produced in the membrane reactor was about 4.5-fold higher (169.2 mg/100 mg) than in ungerminated brown rice (36.82 mg/100 mg), and also higher than that of the other two soaking methods. The γ-oryzanol mass fractions and the antioxidant capacity expressed as ascorbic acid equivalents of germinated brown rice obtained with the three soaking methods varied from 32 to 38 mg/100 mg and 18 to 28 mg/100 g, respectively. Within this study, germination could also slightly reduce the transition temperatures of germinated brown rice starch gelatinization (t o=73-74 °C, t p=76-77 °C and t c=~80 °C, where t o, t p and t c are onset, peak and conclusion (final) temperatures). In conclusion, the production of germinated brown rice in the membrane reactor could enhance its γ-aminobutyric acid mass fraction and reduce wastewater production and is therefore considered more feasible. NOVELTY AND SCIENTIFIC CONTRIBUTION: This study demonstrates the feasibility of germinated brown rice production using a membrane-facilitated soaking reactor with enhancement of bioactive compound content, especially γ-aminobutyric acid, and minimised wastewater production.

Enzymatic Preparation of Bioactive Peptides Exhibiting ACE Inhibitory Activity from Soybean and Velvet Bean: A Systematic Review.

The Angiotensin-I-converting enzyme (ACE) is a peptidase with a significant role in the regulation of blood pressure. Within this work, a systematic review on the enzymatic preparation of Angiotensin-I-Converting Enzyme inhibitory (ACEi) peptides is presented. The systematic review is conducted by following PRISMA guidelines. Soybeans and velvet beans are known to have high protein contents that make them suitable as sources of parent proteins for the production of ACEi peptides. Endopeptidase is commonly used in the preparation of soybean-based ACEi peptides, whereas for velvet bean, a combination of both endo- and exopeptidase is frequently used. Soybean glycinin is the preferred substrate for the preparation of ACEi peptides. It contains proline as one of its major amino acids, which exhibits a potent significance in inhibiting ACE. The best enzymatic treatments for producing ACEi peptides from soybean are as follows: proteolytic activity by Protease P (Amano-P from Aspergillus sp.), a temperature of 37 °C, a reaction time of 18 h, pH 8.2, and an E/S ratio of 2%. On the other hand, the best enzymatic conditions for producing peptide hydrolysates with high ACEi activity are through sequential hydrolytic activity by the combination of pepsin-pancreatic, an E/S ratio for each enzyme is 10%, the temperature and reaction time for each proteolysis are 37 °C and 0.74 h, respectively, pH for pepsin is 2.0, whereas for pancreatin it is 7.0. As an underutilized pulse, the studies on the enzymatic hydrolysis of velvet bean proteins in producing ACEi peptides are limited. Conclusively, the activity of soybean-based ACEi peptides is found to depend on their molecular sizes, the amino acid residues, and positions. Hydrophobic amino acids with nonpolar side chains, positively charged, branched, and cyclic or aromatic residues are generally preferred for ACEi peptides.

Fermented Rice Bran Supplementation Prevents the Development of Intestinal Fibrosis Due to DSS-Induced Inflammation in Mice.

Fermented rice bran (FRB) is known to protect mice intestines against dextran sodium sulfate (DSS)-induced inflammation; however, the restoration of post-colitis intestinal homeostasis using FRB supplementation is currently undocumented. In this study, we observed the effects of dietary FRB supplementation on intestinal restoration and the development of fibrosis after DSS-induced colitis. DSS (1.5%) was introduced in the drinking water of mice for 5 days. Eight mice were sacrificed immediately after the DSS treatment ended. The remaining mice were divided into three groups, comprising the following diets: control, 10% rice bran (RB), and 10% FRB-supplemented. Diet treatment was continued for 2 weeks, after which half the population of mice from each group was sacrificed. The experiment was continued for another 3 weeks before the remaining mice were sacrificed. FRB supplementation could reduce the general observation of colitis and production of intestinal pro-inflammatory cytokines. FRB also increased intestinal mRNA levels of anti-inflammatory cytokine, tight junction, and anti-microbial proteins. Furthermore, FRB supplementation suppressed markers of intestinal fibrosis. This effect might have been achieved via the canonical Smad2/3 activation and the non-canonical pathway of Tgf-ß activity. These results suggest that FRB may be an alternative therapeutic agent against inflammation-induced intestinal fibrosis.

Volatile Compounds, Sensory Profile and Phenolic Compounds in Fermented Rice Bran.

Rice bran (RB), a by-product of the rice milling process, is a rich source of bioactive compounds. Current studies have suggested that fermentation can enhance the bioactivities of RB. This study is aimed to analyse the volatile compounds and sensory profile of fermented RB from two cultivars (Inpari 30 and Cempo Ireng) that are well-known in Indonesia, as well as to measure total phenolic content (TPC) and antioxidant activity. Volatile compounds of fermented RB were analyzed using gas chromatography-mass spectrometry combined with headspace-solid phase microextraction. The optimum TPC and antioxidant activity were observed after 72 h fermentation of RB. The 55 volatile compounds were identified in fermented and non-fermented RB. They were classified into alcohols, aldehydes, acids, ketones, phenols, esters, benzene, terpenes, furans, lactone, pyridines, pyrazines, and thiazoles. Volatile compounds were significantly different among the varieties. The sensory analysis showed that the panelists could differentiate sensory profiles (color, taste, flavor, and texture) between the samples. Fermentation can enhance the acceptance of RB. These studies may provide opportunities to promote the production of fermented RB as a functional ingredient with enhanced bioactivity for health promotion.

The utilization of bilimbi (Averrhoa bilimbi) and lime (Citrus aurantifolia) juices as natural acid coagulants for tofu production.

The influences of coagulation conditions on the characteristics of tofu have been investigated by many studies, with limited perspectives on the utilization of organic acid coagulants. Hence, this research aimed to study the psychochemical and functional properties of tofu coagulated by bilimbi (Averrhoa bilimbi) and lime (Citrus aurantifolia) juices. The highest tofu protein content was quantified for tofu prepared with 20% bilimbi and 5% lime juices, as much as 52.11 and 52.03% (DW), respectively. The corresponding yield was 73.46 and 69.17%. The hardness (155.98 g), gumminess (116.45 g), and chewiness (112.69 g) of treated tofu were found significantly higher than commercial tofu (p < 0.05). Total phenolic content of commercial tofu was about 370.09 µg GAE/g sample (DW). This value was significantly higher than that of treated tofu (p < 0.05). However, the antioxidant activity of the commercial tofu was lower than tofu coagulated with fruit juices. Based on SDS-PAGE analysis, protein band for 11S fraction of tofu coagulated by bilimbi and lime juices were thicker than that of commercial tofu. These small molecular weight peptides might contribute for higher antioxidant activities of tofu coagulated by bilimbi and lime juices. Conclusively, bilimbi and lime juices are potent natural acid coagulants for enhancing the physicochemical and functional properties of tofu.

Supplementation with Fermented Rice Bran Attenuates Muscle Atrophy in a Diabetic Rat Model.

Fermented rice bran (FRB), a prospective supplement, has been proven to ameliorate certain medical conditions. However, its nutraceutical effect on muscle atrophy has never been investigated. The present study aimed to evaluate the effect of FRB on muscle atrophy in a streptozotocin (STZ)-induced diabetic rat model. Three groups of Sprague-Dawley rats, namely the control, STZ, and FRB groups, were treated as follows. The diabetic groups (STZ and FRB) were injected intraperitoneally with STZ (40 mg/kg BW), whereas the control group was injected with the vehicle. The STZ and control groups were fed the AIN93M diet, and the FRB group was fed 10% of FRB based on the AIN93M diet. The diabetic groups had reduced muscle size compared to the control group; however, these changes were alleviated in the FRB group. Moreover, the FRB group had a significantly lower expression of FBXO32/Atrogin-1 and TRIM63/MuRF1 (p < 0.05) due to blocked NF-κB activation. In conclusion, the anti-inflammatory effect of FRB may be beneficial for ameliorating muscle atrophy in diabetic conditions.

Membrane-based preparative methods and bioactivities mapping of tempe-based peptides.

This study was aimed to produce bioactive peptides from optimally fermented tempe, and map their overall bioactivities. There were three preparative methods utilized for producing tempe-based peptides, such as water-facilitated extraction, alcalase, and papain hydrolysis, and in combination with membrane filtration. Fermenting soybean at 144 h was selected as the optimum time based on protein content and degree of hydrolysis. Through SDS-PAGE analysis, an increased degree of hydrolysis with longer fermentation time was confirmed. The best preparative method for producing bioactive peptides was through papain hydrolysis and followed by 5 kDa membrane filtration. By this, the enhancement was distinct for antioxidant activity, ACE-, α-glucosidase-, and Kunitz trypsin-inhibitory activity. The annotated peptide sequences resulting from Nano LC Ultimate 3000 Series System tandem Q Exactive™ Hybrid Quadrupole-Orbitrap™ Mass Spectrometer were matched with the BIOPEP database. The major bioactivities of tempe peptides obtained were as an ACE inhibitor, antioxidant, and antithrombotic.

Rice Bran Reduces Weight Gain and Modulates Lipid Metabolism in Rats with High-Energy-Diet-Induced Obesity.

Obesity has become an epidemic worldwide. It is a complex metabolic disorder associated with many serious complications and high morbidity. Rice bran is a nutrient-dense by product of the rice milling process. Asia has the world's highest rice production (90% of the world's rice production); therefore, rice bran is inexpensive in Asian countries. Moreover, the high nutritional value of the rice bran suggests its potential as a food supplement promoting health improvements, such as enhancing brain function, lowering blood pressure, and regulating pancreatic secretion. The present study evaluated the anti-obesity effect of rice bran in rats with high-energy diet (HED)-induced obesity. Male Sprague-Dawley rats were randomly divided into one of five diet groups (n = 10 per group) and fed the following for eight weeks: Normal diet with vehicle treatment, HED with vehicle, rice bran-0.5X (RB-0.5X) (2% wt/wt rice bran), RB-1.0X (4% wt/wt rice bran), and RB-2.0X (8% wt/wt rice bran). Rice bran (RB-1.0X and RB-2.0X groups) markedly reduced obesity, including body weight and adipocyte size. In addition, treating rats with HED-induced obesity using rice bran significantly reduced the serum uric acid and glucose as well as the liver triglyceride (TG) and total cholesterol (TC). Furthermore, administration of an HED to obese rats significantly affected hepatic lipid homeostasis by increasing phosphotidylcholine (PC; 18:2/22:6), diacylglycerol (DG; 18:2/16:0), DG (18:2/18:1), DG (18:1/16:0), cholesteryl ester (CE; 20:5), CE (28:2), TG (18:0/16:0/18:3), and glycerol-1-2-hexadecanoate 3-octadecanoate. However, the rice bran treatment demonstrated an anti-adiposity effect by partially reducing the HED-induced DG (18:2/18:1) and TG (18:0/16:0/18:3) increases in obese rats. In conclusion, rice bran could act as an anti-obesity supplement in rats, as demonstrated by partially reducing the HED-induced DG and TG increases in obese rats, and thus limit the metabolic diseases associated with obesity and the accumulation of body fat and hepatic lipids in rats.

Dietary Supplementation of Fermented Rice Bran Effectively Alleviates Dextran Sodium Sulfate-Induced Colitis in Mice.

Rice bran (RB) is a major by-product of rice polishing and a rich source of bioactive compounds. Here, we investigated the anti-colitis effect of diet supplementation with fermented rice bran (FRB) in a murine model of ulcerative colitis. FRB was prepared by dual fermentation of RB using fungi and lactic acid bacteria. Colitis was induced in C57Bl/6N male mice (n = 8/group) by dextran sodium sulfate (DSS). Body weight change, disease activity index (DAI), histopathology score, tissue myeloperoxidase (MPO) activity, cytokine and chemokine transcript levels, and the production of short-chain fatty acids (SCFAs) and mucin in the colonic tissue were monitored. Based on histopathology scores, DSS induced severe mucosal inflammation, with an increased loss of crypts, and inflammatory cell infiltration in the control and RB groups, but not in the FRB group. MPO activity, thiobarbituric acid-reactive substance levels, and pro-inflammatory cytokine transcript (Tnf-α, Il-1ß, Il-6, and Il-17) levels were significantly higher in the control and RB groups than in the FRB group. Thus, dietary FRB attenuated intestinal inflammation owing to elevated SCFAs and tryptamine production, which might regulate tight junction barrier integrity and intestinal homeostasis. These results suggest that FRB could comprise an effective potential preventive agent for ulcerative colitis.

Plant Sterol Esters in Extruded Food Model Inhibits Colon Carcinogenesis by Suppressing Inflammation and Stimulating Apoptosis.

Plant sterols in their free forms are known to inhibit colon cancer. Whether these activities persist when compounds are incorporated into processed food is not reported yet. This study aimed to test the ability of plant sterol esters (PSE) incorporated into a nonpuffed extruded food (NPE) model to inhibit colon carcinogenesis. PSE was added into NPE at four concentrations (0.0%, 0.7%, 1.4%, and 2.1%). PSE-NPE activity was tested in azoxymethane/dextran sodium sulfate-induced Balb/c mice. The groups given PSE-NPE did not show any colon tumor formation. Immunohistochemistry results revealed that the group fed with 1.4% PSE had the lowest histoscore for cyclooxygenase-2 expression and the highest histoscore for cleaved caspase-3, cleaved caspase-8, and cleaved caspase-9expressions. The results of this study indicated that even after incorporation into a food system, which is processed using high pressure and temperature, PSE retained its chemopreventive activity. The proposed mechanisms are by suppressing inflammation and inducing apoptosis.

Fermented rice bran supplementation mitigates metabolic syndrome in stroke-prone spontaneously hypertensive rats.

BACKGROUND: Previous study shown that enzyme treated-rice bran effectively improved hypertension and glucose intolerance in stroke-prone spontaneously hypertensive rat (SHRSP). However, dual fermentation of rice bran's efficacy against metabolic syndrome in SHRSP is still unknown. METHODS: Fermented rice bran (FRB) was prepared by dual fermentation of rice bran using fungi and lactic acid bacteria. The effect of FRB on metabolic syndrome in stroke-prone spontaneously hypertensive rats (SHRSP) was investigated by single and chronic supplementation. RESULTS: Dual fermentation of rice bran enriches the functional value of rice bran. Single-dose oral administration of FRB (2 g/kg body weight) reduced systolic blood pressure; however, chronic supplementation with 5 % FRB (4 weeks) significantly reduced both systolic and diastolic blood pressure. FRB supplementation improved leptin impairment and increased serum adiponectin levels and angiotensin-converting enzyme inhibitory activity. Furthermore, FRB supplementation improved glucose tolerance and insulin sensitivity as well as serum insulin levels. Lipid profiles were also improved by the regulation of 5' adenosine monophosphate-activated protein kinase activation. Moreover, supplementation with FRB reduced the expressions of hepatic transcription factors such as liver X receptor alpha, sterol regulatory element-binding protein 1c, and carbohydrate-responsive element-binding protein alpha, as well as their target genes. In conclusion, dietary supplementation with FRB may lower hypertension and alleviate metabolic syndrome. CONCLUSION: Metabolic syndrome was better alleviated with FRB supplementation. We therefore suggest FRB as an alternative medicine to reduce the risks of lifestyle-related diseases.

NMR metabolomics for identification of adenosine A1 receptor binding compounds from Boesenbergia rotunda rhizomes extract.

ETHNOPHARMACOLOGICAL RELEVANCE: Boesenbergia rotunda Linn. (Zingiberaceae) is traditionally used in many Asian countries as medicine for stomach pain and discomfort, viral and bacterial infection, inflammation, and as diuretic agent. AIM OF THE STUDY: The study aimed to identify adenosine A1 receptor binding compounds from Boesenbergia rotunda rhizome extract by using comprehensive extraction coupled to the NMR metabolomics method. MATERIALS AND METHODS: Dried and powdered Boesenbergia rotunda rhizomes were extracted with the comprehensive extraction method to obtain several fractions with different polarity. Each fraction was divided into two: for NMR analysis and for adenosine A1 receptor binding test. Orthogonal projection to the least square analysis (OPLS) was used to study the correlation between metabolites profile and adenosine A1 receptor binding activity of the plant extracts. Based on Y-related coefficient and variable of important (VIP) value, signals in active area of OPLS loading plot were studied and the respective compounds were then elucidated RESULTS AND DISCUSSIONS: Based on OPLS Y-related coefficient plot and variable of importance value plot, several characteristic signals were found to positively correlate to the binding activity. By using 1D and 2D NMR spectra of one of the most active fraction, pinocembrine and hydroxy-panduratin were identified as the possible active compounds. Two signals from ring C of pinocembrine flavanone skeleton with negative coefficient correlations possibly overlapped with those of non-active methoxylated flavanones which were also presence in the extract. NMR based metabolomics applied in this study was able to quickly identify bioactive compounds from plant extract without necessity to purify them. Further confirmation by isolating pinocembrine and hydroxy-panduratin and testing their adenosine A1 receptor binding activity to chemically validate the method are required. CONCLUSION: Two flavonoid derivatives, pinocembrine and hydroxy-panduratin, have been elucidated as possible active compounds bind to adenosine A1 receptor. Flavonoid was reported to be one of natural antagonist ligand for adenosine A1 receptor while antagonistic activity to the receptor is known to associate with diuretic activity. Thus, the result of this research supports the traditional use of Boesenbergia rotunda rhizome extract as diuretic agent.