Testa Extracts Research Articles

Enzyme‑assisted extraction of cashew nut (Anacardium occidentale L.) testa

Enzyme-assisted extraction (EAE) has been successfully applied to extract polyphenols from various food materials and by-products. Hence, the present research aimed to valorize the cashew nut (Anacardium occidentale L.) testa by EAE method using an enzyme cocktail of cellulase and pectinase. The effects of different processing parameters, including the pH (3.0 - 6.0), temperature (30 - 70°C), extraction time (60 - 150 min), solid -to-solvent ratio (1/5 - 1/85), and enzyme concentration (0.1 - 0.4% v/v) on the total polyphenol content (TPC) and antioxidant capacity of the testa extracts were investigated. The results showed that the TPC of the testa extracts positively correlated with the antioxidant capacity. The EAE performed at pH 4.0, a temperature of 50°C, an extraction time of 60 mins, a solid-to-solvent ratio of 1/55, and an enzyme concentration of 0.2% (v/ v) resulted in the highest TPC (168.84 g GAE/kg DW) with DPPH and ABTS antioxidant activity of 885.56 and 1513.60 (mmol TE/kg DW), respectively. The EAE technique holds significant promise as a sustainable approach to mitigate the underutilization and undervaluation of cashew nut testa waste, which could otherwise contribute to environmental pollution.

The Effects of C3G and D3G Anthocyanin-Rich Black Soybean on Energy Metabolism in Beige-like Adipocytes.

Various mechanisms of obesity prevention have been identified; however, the roles of brown or beige fat as regulators of the energy balance are unclear. The effects of anthocyanin-rich black soybean, Glycine max (L.) Merr., testa (ABS) extracts on the energy balance were investigated by comparing beige-like adipocytes (BLA) and white adipocytes (WAT). ABS extracts reduced peroxisome proliferator-activated receptor gamma protein expression and triglyceride accumulation in WAT and BLA without inducing nuclear damage. The biomarkers of fat degradation (phospho-AMPKα and ATGL) or glycerol secretion in the medium and β-oxidation of fatty acids (CPT2) in the ABS-treated BLA were increased compared to those in WAT. ABS extracts significantly increased the expression of thermogenesis markers (UCP1 and CIDEA) and biomarkers related to mitochondrial activation (cytochrome c and NRF1) in BLA. In the primary cell culture of brown adipocytes (BAT) from rats fed ABS, the expression levels of PGC1-α, cytochrome c, and UCP1 proteins were increased compared to those in BAT from nonfed rats. A reduction in the NAD/NADH ratio was consistently associated with an increase in the oxygen consumption rate and basal/maximal respiration rate in ABS-treated BLA. Anthocyanins promote beiging in the body, contribute to the prevention of obesity, and are potentially useful functional materials.

Peanut testa extracts enhance anticancer effect of cisplatin against human cholangiocarcinoma cells via modulation of histone deacetylase inhibitory activity

Objective: To investigate the effect of combination treatments of cisplatin and KK4 and ICG15042 peanut testa extracts against cholangiocarcinoma cells in vitro. Methods: The growth inhibition, cell cycle arrest and apoptosis of cholangiocarcinoma cells were determined by 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and flow cytometry analysis, respectively. The levels of proteins involved in apoptosis were assessed using Western blotting assays. The caspase activity was assessed using a colorimetric caspase activity assay. Results: Cisplatin and peanut (KK4 and ICG15042) testa extracts inhibited the growth of cholangiocarcinoma cell lines (KKU- M214 and KKU-100 cells) in a dose- and time-dependent manner. The combination treatments reduced cell viability and induced apoptosis of cholangiocarcinoma cells more efficiently than singledrug treatments. Cancer cell death synergistically mediated by cisplatin and peanut testa extracts was observed in KKU-M214 cells (combination index 1.0). The combination treatments also increased the sub- G1 population and caused KKU-M214 cell cycle arrest at S and G2/ M phases, which were the combined effects of cisplatin (S phase arrest) and peanut testa extracts (G2/M phase arrest). In addition, pERK1/2, Ac-H3, Bcl-2 and proteins related to apoptosis, including Bax and caspases 3, 8, 9, exhibited enhanced expression in KKU- M214 cells. The combination treatments caused down-regulation of p53, whereas the expression of p21 was fairly constant when compared with cisplatin single drug treatment. Conclusions: Peanut testa extracts in combination with cisplatin synergistically reduce cell viability and induce apoptosis through stimulation of caspases 3, 8 and 9 in KKU-M214 cells.

In vitro anti-Helicobacter pylori activity of aqueous extract from Persian Oak testa

ObjectiveGrowing problem of antibiotic resistance in Helicobacter pylori, as a common cause of chronic gastritis and even stomach cancer, demands searching for novel candidates of herbal sources. This study is aimed at assessing the antimicrobial activity of aqueous extract obtained from Quercus brantii var. persica seed coat (Testa) on H. pylori isolated from gastric biopsy specimens. MethodsSuch specimens were collected from 100 patients presenting with endoscopic gastroduodenal findings. Testa extracts were prepared from Persian Oak forests in the province of Kohgiluyeh and Boyer-Ahmad, IRAN. H. pylori isolates were obtained by a series of standard bacteriology tests and cell culture, then were confirmed by PCR. The activity of testa extracts towards 25 H. pylori isolates was assessed by well diffusion method, microdilution assay, and a disk diffusion assay in vitro. Results were analyzed statistically by one-way ANOVA analysis. ResultsAqueous extract of testa demonstrated an antimicrobial activity with zone diameters of inhibition ranged from 0 mm to 40 mm. Its inhibitory activity increased simultaneously with increasing extract concentration. The lowest MIC and MBC were both recorded as 2 µg/mL. Anti-H. pylori activity of testa extract was approximately close to tetracycline and metronidazole and less than amoxicillin. A potent extract of testa possessed significant inhibitory activity (P < 0.05). ConclusionTesta extract is suggested as a natural therapeutic source against the gastric H. pylori infection. However, evaluating the in vivo activity of this extract is necessary too.

Peanut testa extracts possessing histone deacetylase inhibitory activity induce apoptosis in cholangiocarcinoma cells

Previous studies demonstrated that peanut testa extracts (KK4 and ICG15042) containing natural histone deacetylase (HDAC) inhibitors inhibited the growth of several human cancer cell lines via apoptosis induction. The aims of this study were to investigate the anti-proliferative effects and the mechanism(s) responsible for apoptosis induction mediated by these peanut testa extracts in human cholangiocarcinoma cell lines (KKU-M214 and KKU-100). The anti-proliferative effects were assessed by MTT assay. Apoptotic cell death and cell cycle arrest were analyzed by flow cytometry. The caspase activities were studied using colorimetric caspase activity assay and western blot analysis. Our results revealed that KK4 and ICG15042 extracts inhibited cell proliferation of both KKU-M214 and KKU-100 cells in a dose- and time-dependent manner, with IC50 values of 38.28 ± 0.29 (KK4), 43.91 ± 1.94 (ICG15042) μg/mL for KKU-M214 and 78.40 ± 1.74 (KK4), 82.77 ± 0.94 (ICG15042) μg/mL for KKU-100 at 72 h. Apoptosis induction by these peanut testa extracts were observed in both KKU-M214 and KKU-100 cells in a concentration-dependent manner. Moreover, the percentage of cells in the sub-G1 phase was significantly increased in both KKU-M214 and KKU-100 cells. Cell cycle arrest was not observed in other cell cycle phases. Activation of caspases 8 and 3 were apparent integral parts of apoptosis induction in both cells. Both peanut testa extracts also caused down-regulation of p53, p21, Bcl-2 and pERK1/2 protein expression in these cells. These results suggest that peanut testa extracts may be potential anti-cancer agents for cholangiocarcinoma chemoprevention or chemotherapy.

Anthocyanin Rich-Black Soybean Testa Improved Visceral Fat and Plasma Lipid Profiles in Overweight/Obese Korean Adults: A Randomized Controlled Trial.

Anthocyanin-rich black soybeans have been used in traditional East Asian medicine to cure diseases related to oxidative stress and carcinogens, but not obesity. Our objective was to investigate the effects of anthocyanin-rich black soybean testa extracts (BBT), Glycine max (Chongja No. 3), on obesity. In total, 63 participants defined as overweight or obese by their body mass index (BMI >23) or waist circumference (WC >90 cm for males, >85 cm for females) were sorted into two groups: 32 receiving the trial medication (BBT, 2.5 g/d) and 31 receiving the placebo (starch, 2.5 g/d). Participants completed an 8-week, randomized, double-blinded, and placebo-controlled clinical trial. There were no significant differences between the two groups at the beginning of the trial, and both required the same safety assessments. Significant decreases in abdominal fat, described according to WC and hip circumference, and lipid profiles such as triacylglycerols (TG), low density lipoprotein cholesterol (LDLc), and non-high density lipoprotein cholesterol (non-HDLc) were observed in the BBT group at the conclusion of the clinical trial. The indicators for arteriosclerosis such as total cholesterol (TC)/HDLc and LDLc/HDLc were significantly decreased in the BBT group, but had not changed in the placebo group. With no difference between the two groups in energy-adjusted dietary intakes and physical activity, BBT was shown to strongly improve plasma lipid profiles, related to the reduction of WC (an indicator of abdominal fat) as long as high dietary fiber and low cholesterol diets were maintained. In conclusion, BBT can potentially be developed as a functional food for preventing abdominal obesity with high fiber and low cholesterol diets.

PHYTOCHEMICALS AND ANTIOXIDANT ACTIVITY OF TESTA EXTRACTS OF COMMERCIAL WET AND DRY COCONUTS AND CAKES

Coconut testa is the brown colored outer thin layer covering the coconut kernel. It is a by-product obtained in the coconut processing industries through paring of wet coconuts to make products like virgin coconut oil, snow ball coconut, desiccated coconut and coconut milk. A study was carried out for the phytochemical screening and antioxidant activity of testa extracts. The wet coconut testa (WCT), copra testa (CT), copra cake (CC), wet coconut whole (WCW) and copra whole (CW) were investigated for the presence of phytochemicals and antioxidants. The extracts were found to contain carbohydrates, amino acids, glycosides, triterpenes, tannins, flavonoids, phenolics, and saponins. The IC50values (0.06-0.5 mg/ml) and reducing power were (0.5-9 g equivalent of TBHQ/100 g) of extracts indicate its antioxidant potential. The extracts contained phenolic acids, flavonoids, and total tocopherols. These studies indicated that the extracts from wet coconut testa had high antioxidant activity and contained many phytochemicals when compared to other coconut testa extracts.

Histone Deacetylase Inhibitory Activity of Peanut Testa Extracts against Human Cancer Cell Lines

Inhibition of histone deacetylase (HDAC) is a promising strategy for cancer treatment. In this study, HDAC inhibitory activity of 15 Valencia-type peanut testa extracts was assessed in mammalian cell model. Nine of 15 testa extracts exhibited HDAC inhibitory activity. Two peanut testa extracts (genotypes ICG15042 and KK4) possessing the greatest HDAC inhibitory activity could inhibit the growth of all five human cancer cell lines tested. At 72-h treatment, both ICG15042 and KK4 testa extracts showed the most effective growth inhibition on Jurkat T-leukemia cells with IC50 values of 28.81 and 30.00 μg/mL, respectively. Both extracts were more toxic against the cancerous cell lines, but safer toward a noncancer cell line. Growth inhibitory effects of both extracts appeared to be mediated by induction of apoptosis in a dose-dependent manner. Moreover, some phenolic acids including protocatechuic, syringic, p-coumaric, ferulic and sinapinic acids, which may underpin their anticancer activity, were also identified and analyzed quantitatively. Practical Applications HDAC inhibitory potentials of phenolic-rich testa extracts of 15 Valencia-type peanut genotypes in mammalian cell model were evaluated. The results suggest that peanut skins (testae) of nine peanut genotypes possessed HDAC inhibitory activity and two of them (ICG15042 and KK4), with the highest HDAC inhibitory activity, could inhibit cancer cell growth via apoptosis induction. Accordingly, peanut skins could be useful for application in areas such as alternative medicine for cancer treatment and functional food for dietary prevention of cancer.

Effects of the C3G/D3G anthocyanins-rich black soybean testa extracts on improvement of lipid profiles in STZ-induced diabetic rats

Purpose: The purpose of this study was to examine the effect of black soybean (CJ-3) testa extracts on lipid profiles in streptozotocin (STZ)-induced diabetic rats. Methods: One control group and four STZ-induced diabetic groups with different doses of black soybean (CJ-3) testa extracts treatment [0 mg/kg (diabetic control, EX), 250 mg/kg (EX-250), 500 mg/kg (EX-500), 1,000 mg/kg (EX-1000)] were orally administered for 4 weeks. Results: All CJ-3 treatment groups had remarkably lower serum triglyceride (TG) levels than that of EX group (p < 0.05) whereas hepatic TG contents did not show any differences. Results from serum total cholesterol (TC) concentrations of EX-250 and EX-1000 groups were decreased compared to EX group (p < 0.05). Furthermore, protein levels of 3-hydroxy-3-methyl-glutaryl-Coenzyme A (HMG-CoA) reductase from the liver decreased in all treatment groups (p < 0.05). However, significant differences were not observed in serum glucose and insulin, and glucose transporter 4 (GLUT-4) protein expression in skeletal muscle tissue. Conclusion: These results suggest that black soybean testa extracts could be useful for improvement of hyperlipidemia and hypercholesteremia in diabetes.

Antioxidative potential of cashew phenolics in food and biological model systems as affected by roasting

The effect of different roasting conditions on antioxidant capacity of phenolics of cashew nuts and their testa was evaluated using several food and biological model systems. Total phenolic content (TPC) of cashew extracts was determined and accelerated oxidative stability of stripped corn oil in the presence of cashew extracts evaluated. In addition, the antioxidant activity of the extracts was assessed in a β-carotene-linoleate and a cooked comminuted pork model system. Inhibition of oxidation of human low density lipoprotein (LDL) cholesterol and stand breaking of supercoiled deoxyribonucleic acid (DNA) was also investigated. The TPC ranged from 5 to 791mg gallic acid equivalents/g crude extract. In general, whole cashew nuts and testa extracts demonstrated stronger antioxidant activity than that of the cashew kernel. The inhibition percentage of LDL cholesterol oxidation, as evaluated by conjugated dines formation, of cashew kernels was higher than that of testa and was 69% at the end of 24h incubation. Extracts of roasted cashew nut showed considerable antioxidative efficiency in model systems employed in this study, however, the effect was not significantly (P⩽0.05) different from that of their raw counterparts, except for the accelerated oxidative stability assay. The results suggest that whole cashew nut and testa extracts could be used as a potential source of natural antioxidants in certain food applications and for disease risk reduction.

The effect of sesbanimide and Sesbania seed extracts on germination and seedling growth of a number of plant species

Sesbanimide and extracts made from ground powders of whole Sesbania punicea and S. bispinosa seeds and isolated embryos and testae of S. punicea inhibited the germination of vegetable, grass and weed seeds. This inhibition was more noticeable on seedling growth than on germination. The inhibitory effect was not transitory but was manifested even after transplanting the seedlings. In all cases, the roots were more sensitive than the shoots and their growth was severely retarded. The inhibitory effect was also detected if the test seeds were soaked in extracts prior to sowing. Embryo and testa extracts of S. punicea gave an inhibitory response. The former was the most inhibitory. This inhibition may partly explain why S. punicea forms dense, uniform stands when invading natural vegetation. There appears to be a strong allelopathic effect expressed by the seed extracts. This warrants further investigation.

Alfalfa mosaic virus in lucerne seed during seed maturation and storage, and in seedlings

The incidence of alfalfa mosaic virus (AMV) in lucerne seed and pods during maturation, when monitored by sap transmission to Phaseolus (infective virus) and ELISA (AMV antigen), showed that infective virus incidence decreased rapidly with maturation, whereas antigen incidence declined slowly and was always higher than infective virus. Infective virus and antigen incidence were higher in mature seed of cv. Maris Kabul than cv. Europe because virus inactivation/degradation were more rapid in cv. Europe. Seed infection with virus originating from pollen, ovules or both was found in pods and seeds 12–15 days after pollination between healthy or AMV‐infected plants; this was before maturation‐associated virus inactivation. Ovule transmission was more frequent than pollen transmission. AMV antigen was present in embryos and testas of mature seed; infective virus only in embryos. Non‐infective but ELISA‐positive antigen in testa extracts accounted for the higher incidence of ‘seed‐borne AMV’ compared with embryo‐associated seed transmission to seedlings. Tests with dry mature seed either underestimated (infectivity tests) or overestimated (ELISA) eventual seedling infection. Infectivity and ELISA tests gave identical incidence values for 17 to 29‐day‐old seedlings.

The phytate content and iron‐binding capacity of various field bean (Vicia faba) preparations and extracts

AbstractSeed coats, cotyledons and whole grains from a number of field bean (Vicia faba) varieties were analysed for phytate content. In the case of the white flowered varieties, phytate content and its associated iron‐binding capacity appeared to be restricted to the cotyledons while seed coat extracts prepared from coloured flowered varieties had almost twice the iron‐binding capacity of similarly prepared cotyledon extracts. Further studies of these testa extracts revealed that this iron‐binding capacity was eliminated in the presence of either excess protein or polyvinyl pyrrolidone and it was concluded that the active constituents in the seed coats of coloured flowered varieties were the condensed tannins. The solubility and precipitation characteristics at different pH values of field bean phytate were also determined and although the points of minimum solubility broadly coincided with that of field bean protein, phytate solubility increased more rapidly thereafter with increasing pH. These differences in solubility facilitated the preparation of a range of protein isolates with phytate contents ranging from 1.08% at pH 6 to 6.13% at pH 3.