The Olive Biophenols Oleuropein and Hydroxytyrosol Selectively Reduce Proliferation, Influence the Cell Cycle, and Induce Apoptosis in Pancreatic Cancer Cells.

Current chemotherapy drugs for pancreatic cancer only offer an increase in survival of up to six months. Additionally, they are highly toxic to normal tissues, drastically affecting the quality of life of patients. Therefore, the search for novel agents, which induce apoptosis in cancer cells while displaying limited toxicity towards normal cells, is paramount. The olive biophenols, oleuropein, hydroxytyrosol and tyrosol, have displayed cytotoxicity towards cancer cells without affecting non-tumorigenic cells in cancers of the breast and prostate. However, their activity in pancreatic cancer has not been investigated. Therefore, the aim of this study was to determine the anti-pancreatic cancer potential of oleuropein, hydroxytyrosol and tyrosol. Pancreatic cancer cells (MIA PaCa-2, BxPC-3, and CFPAC-1) and non-tumorigenic pancreas cells (HPDE) were treated with oleuropein, hydroxytyrosol and tyrosol to determine their effect on cell viability. Oleuropein displayed selective toxicity towards MIA PaCa-2 cells and hydroxytyrosol towards MIA PaCa-2 and HPDE cells. Subsequent analysis of Bcl-2 family proteins and caspase 3/7 activation determined that oleuropein and hydroxytyrosol induced apoptosis in MIA PaCa-2 cells, while oleuropein displayed a protective effect on HPDE cells. Gene expression analysis revealed putative mechanisms of action, which suggested that c-Jun and c-Fos are involved in oleuropein and hydroxytyrosol induced apoptosis of MIA PaCa-2 cells.

Development of an objective measure of quality and commercial value of Japanese-styled green tea (Camellia L. sinensis): the Quality Index Tool.

A novel approach to evaluate the commercial value of green tea products is explored in this paper. The green tea Quality Index Tool (QI-Tool) is based on high performance liquid chromatography (HPLC), capable of identifying and understanding the constituents that are important to create superior consumer and commercially valuable green tea beverages in the Japanese-style. This tool will allow producers to better identify a product's potential value within the various levels of green tea retail quality structure. Via the quantification of theanine, caffeine and the catechins: epicatechin (EC), epicatechin gallate (ECG), epigallocatchin (EGC), epigallocatechin gallate (EGCG) and gallocatechin gallate (GCG) within a green tea beverage, the QI-Tool provides categorisation of a product against the green tea market retail competitive set. This allows a better understanding of the product's potential commercial value, as well as a comparison to other products within that market category. The QI-Tool is an alternative and promising method for objectively evaluating commercial value of green tea products using HPLC analysis.

Effects of four different drying methods on the carotenoid composition and antioxidant capacity of dried Gac peel.

BACKGROUND: Gac fruit (Momordica cochinchinensis Spreng.) is a rich source of carotenoids for the manufacture of powder, oil and capsules for food, cosmetic and pharmaceutical uses. Currently, only the aril of the Gac fruit is processed and the peel, similar to the other components, is discarded, although it contains high level of carotenoids, which could be extracted for commercial use. In the present study, four different drying methods (hot-air, vacuum, heat pump and freeze drying), different temperatures and drying times were investigated for producing dried Gac peel suitable for carotenoid extraction. RESULTS: The drying methods and drying temperatures significantly affected the drying time, carotenoid content and antioxidant capacity of the dried Gac peel. Among the investigated drying methods, hot-air drying at 80 o C and vacuum drying at 50 o C produced dried Gac peel that exhibited the highest retention of carotenoids and the strongest antioxidant capacity. CONCLUSION: Hot-air drying at 80 o C and vacuum drying at 50 o C are recommended for the drying of Gac peel. © 2016 Society of Chemical Industry.

Phytochemical properties and anti-proliferative activity of Olea europaea L. leaf extracts against pancreatic cancer cells.

Olea europaea L. leaves are an agricultural waste product with a high concentration of phenolic compounds; especially oleuropein. Oleuropein has been shown to exhibit anti-proliferative activity against a number of cancer types. However, they have not been tested against pancreatic cancer, the fifth leading cause of cancer related death in Western countries. Therefore, water, 50% ethanol and 50% methanol extracts of Corregiola and Frantoio variety Olea europaea L. leaves were investigated for their total phenolic compounds, total flavonoids and oleuropein content, antioxidant capacity and anti-proliferative activity against MiaPaCa-2 pancreatic cancer cells. The extracts only had slight differences in their phytochemical properties, and at 100 and 200 µg/mL, all decreased the viability of the pancreatic cancer cells relative to controls. At 50 µg/mL, the water extract from the Corregiola leaves exhibited the highest anti-proliferative activity with the effect possibly due to early eluting HPLC peaks. For this reason, olive leaf extracts warrant further investigation into their potential anti-pancreatic cancer benefits.

Greenhouse-grown bitter melon: production and quality characteristics.

BACKGROUND: Bitter melon (Momordica charantia L.) is a medicinal fruit reported to have antidiabetic properties. To grow this tropical fruit year-round in temperate climates, greenhouse production is necessary, sometimes without insect pollinators. Suitable high-yielding varieties with good bioactivity need to be identified. This experiment evaluated the yield of six varieties of bitter melon under greenhouse conditions and their bioactivity in terms of total phenolic and saponin compounds and total antioxidant activity determined using four assays. RESULTS: The larger varieties (Big Top Medium, Hanuman, Jade and White) were more productive than the small varieties (Indra and Niddhi) in terms of total fruit weight and yield per flower pollinated. The bioactivity (total phenolic and saponin compounds and antioxidant activity) of the two small varieties and Big Top Medium was significantly higher than that of the other three large varieties. Two antioxidant assays, 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric-reducing antioxidant power (FRAP), were shown to provide the strongest correlations with phenolic and saponin compounds of bitter melon. CONCLUSION: Preliminary research has identified Big Top Medium as the most suitable variety for greenhouse production. The rich source of phenolic and saponin compounds and their associated antioxidant activity highlight bitter melon as a valuable food.

Turmeric (Curcuma longa L.) drying: an optimization approach using microwave-vacuum drying.

This study investigated the effect of microwave power (2,400-4,000 W) and drying times (10-30 min) on the quality of dried turmeric in terms of colour (L, a*, b*), moisture content, water activity (aw), ash, antioxidant activity (2,2-dipheneyl-1-picrylhydrazyl; DPPH), total phenolic and curcuminoids content. In addition, Response Surface Methodology (RSM) was implemented to optimize the drying conditions for turmeric. A range of quality parameters, microwave power (W) and time (min) were fitted to quadratic models using a central composite design. The Analysis of Variance (ANOVA) statistic results suggested that the optimal qualities (i.e., aw, value, moisture content, phenolic content, curcuminoid content and EC50 and L value) of dried turmeric were achieved at high vacuum-microwave power (3,500-4,000 W) and long duration (27-30 min). The improvement in the quality of dried turmeric microwave-vacuum drying in these conditions was illustrated through the enzymatic browning reaction via inhibition of polyphenol oxidase which suppressed the formation of the brown pigments and increased the phenol substrates.

Optimum conditions for the water extraction of L-theanine from green tea.

Theanine is a unique non-protein amino acid found in tea (Camellia sinensis). It contributes to the favourable umami taste of tea and is linked to various beneficial effects in humans. There is an increasing interest in theanine as an important component of tea, as an ingredient for novel functional foods and as a dietary supplement. Therefore, optimal conditions for extracting theanine from tea are required for the accurate quantification of theanine in tea and as an efficient first step for its purification. This study examined the effects of four different extraction conditions on the yield of theanine from green tea using water and applied response surface methodology to further optimise the extraction conditions. The results showed that temperature, extraction time, ratio of water-to-tea and tea particle sizes had significant impacts on the extraction yield of theanine. The optimal conditions for extracting theanine from green tea using water were found to be extraction at 80 °C for 30 min with a water-to-tea ratio of 20:1 mL/g and a tea particle size of 0.5-1 mm.

Optimizing conditions for the extraction of catechins from green tea using hot water.

Six different factors involved in the extraction of catechins from green tea using water were examined for their impact on the yield of catechins and on the efficiency of water use. The best temperature and time combination for catechin extraction was at 80°C for 30 min. The yield of catechins was also optimal with a tea particle size of 1 mm, a brewing solution pH <6 and a tea-to-water ratio at 50:1 (mL/g). In terms of efficient use of water in a single extraction, a water-to-tea ratio of 20:1 (mL/g) gave the best results; 2.5 times less water was used per gram of green tea. At the water-to-tea ratio of 20:1 mL/g, the highest yield of catechins per gram of green tea was achieved by extracting the same sample of green tea twice. However, for the most efficient use of water, the best extraction was found to be once at a water-to-tea ratio of 12:1 (mL/g) and once at a water-to-tea ratio of 8:1 (mL/g). Therefore, all six of the factors investigated had an impact on the yield of catechins extracted from green tea using water and two had an impact on the efficiency of water use.

L-Theanine: properties, synthesis and isolation from tea.

Theanine is a non-protein amino acid that occurs naturally in the tea plant (Camellia sinensis) and contributes to the favourable taste of tea. It is also associated with effects such as the enhancement of relaxation and the improvement of concentration and learning ability. It is also linked with health benefits including the prevention of certain cancers and cardiovascular disease, the promotion of weight loss and enhanced performance of the immune system. Thus, there has been a significant rise in the demand for theanine. While theanine has been chemically and biologically synthesised, techniques to isolate theanine from natural sources remain an important area of research. In this review article, the properties and health benefits of theanine are summarised and the synthesis and isolation of theanine are reviewed and discussed. Future perspectives for the isolation of theanine from natural sources are also outlined.

Extraction and isolation of catechins from tea.

Tea is a major source of catechins, which have become well known for their antioxidant potential. Numerous human, animal, and in vitro studies have linked tea catechins with prevention of certain types of cancers, reduction of the risks for obesity, diabetes, and cardiovascular disease, and improvement of the immune system. Tea catechins are widely used in various neutraceuticals, pharmaceuticals, and cosmetics for either enhancing product shelf-life or for enhancing human health. Thus, the demand for catechins has increased considerably. Catechins have been extracted and isolated from tea leaves by numerous methods through several steps including: treatment of the tea leaves, extraction of catechins from teas into solvents, isolation of catechins from other extracted components, and drying the preparations to obtain catechin extracts in a powder form. This paper outlines the physical and chemical properties of the tea catechins and reviews the extraction steps of the various extraction methods, as a basis to improve and further develop the extraction and isolation of the tea catechins.

Relationship Between B-Vitamin Biomarkers and Dietary Intake with Apolipoprotein E є4 in Alzheimer's Disease.

The potential for B-vitamins to reduce plasma homocysteine (Hcy) and reduce the risk of Alzheimer's disease (AD) has been described previously. However, the role of Apolipoprotein E є4 (APOE4) in this relationship has not been adequately addressed. This case-control study explored APOE4 genotype in an Australian sample of 63 healthy individuals (female = 38; age = 76.9 ± 4.7 y) and 63 individuals with AD (female = 35, age = 77.1 ± 5.3 y). Findings revealed 55 of 126 participants expressed the APOE4 genotype with 37 of 126 having both AD and the APOE4 genotype. Analysis revealed an increased likelihood of AD when Hcy levels are >11.0 µmol/L (p = 0.012), cysteine levels were <255 µmol/L (p = 0.033) and serum folate was <22.0 nmol/L (p = 0.003; in males only). In females, dietary intake of total folate <336 µg/day (p=0.001), natural folate <270 µg/day (p = 0.011), and vitamin B2 < 1.12 mg/day (p = 0.028) was associated with an increased AD risk. These results support Hcy, Cys, and SF as useful biomarkers for AD, irrespective of APOE4 genotype and as such should be considered as part of screening and managing risk of AD.

Optimisation of the Microwave-Assisted Ethanol Extraction of Saponins from Gac (Momordica cochinchinensis Spreng.) Seeds.

BACKGROUND: Gac (Momordica cochinchinensis Spreng.) seeds contain saponins that are reportedly medicinal. It was hypothesised that the extraction of saponins from powdered Gac seed kernels could be optimised using microwave-assisted extraction (MAE) with ethanol as the extraction solvent. The aim was to determine an appropriate ethanol concentration, ratio of solvent to seed powder and microwave power and time for extraction. Whether or not defatting the Gac seed powder had an impact on the extraction of saponins, was also determined. RESULTS: A four-fold higher total saponin content (TSC) was obtained in extracts from full-fat Gac seed kernel powder than from defatted powder (100 vs. 26 mg aescin equivalents (AE) per gram of Gac seeds). The optimal parameters for the extraction of saponins were a ratio of 30 mL of 100% absolute ethanol per g of full-fat Gac seed kernel powder with the microwave set at 360 W for three irradiation cycles of 10 s power ON and 15 s power OFF per cycle. CONCLUSIONS: Gac seed saponins could be efficiently extracted using MAE. Full-fat powder of the seed kernels is recommended to be used for a better yield of saponins. The optimised MAE conditions are recommended for the extraction of enriched saponins from Gac seeds for potential application in the nutraceutical and pharmaceutical industries.

Bioactive Composition, Antioxidant Activity, and Anticancer Potential of Freeze-Dried Extracts from Defatted Gac (Momordica cochinchinensis Spreng) Seeds.

Background: Gac (Momordica cochinchinensis Spreng) seeds have long been used in traditional medicine as a remedy for numerous conditions due to a range of bioactive compounds. This study investigated the solvent extraction of compounds that could be responsible for antioxidant activity and anticancer potential. Methods: Defatted Gac seed kernel powder was extracted with different solvents: 100% water, 50% methanol:water, 70% ethanol:water, water saturated butanol, 100% methanol, and 100% ethanol. Trypsin inhibitors, saponins, phenolics, and antioxidant activity using the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and the ferric reducing antioxidant power (FRAP) assays; and anticancer potential against two melanoma cancer cell lines (MM418C1 and D24) were analysed to determine the best extraction solvents. Results: Water was best for extracting trypsin inhibitors (581.4 ± 18.5 mg trypsin/mg) and reducing the viability of MM418C1 and D24 melanoma cells (75.5 ± 1.3 and 66.9 ± 2.2%, respectively); the anticancer potential against the MM418C1 cells was highly correlated with trypsin inhibitors (r = 0.92, p < 0.05), but there was no correlation between anticancer potential and antioxidant activity. The water saturated butanol had the highest saponins (71.8 ± 4.31 mg aescin equivalents/g), phenolic compounds (20.4 ± 0.86 mg gallic acid equivalents/g), and antioxidant activity, but these measures were not related to anticancer potential. Conclusions: Water yielded a Gac seed extract, rich in trypsin inhibitors, which had high anticancer potential against two melanoma cell lines.

Microwave-assisted extraction and ultrasound-assisted extraction for recovering carotenoids from Gac peel and their effects on antioxidant capacity of the extracts.

The peel of Gac fruit (Momordica cochinchinensis Spreng.) contains high levels of bioactive compounds, especially carotenoids which possess significant antioxidant capacities. However, the peel of Gac is regarded as a waste from the production of carotenoid-rich oil from Gac fruit. In this study, carotenoids of Gac peel were extracted by microwave-assisted extraction (MAE) and ultrasound-assisted extraction (UAE) using ethyl acetate as extraction solvent. The effect of extraction time and different levels of microwave and ultrasonic powers on the yield of total carotenoid and antioxidant capacity of the extracts were investigated. The results showed that an extraction at 120 W for 25 min and an extraction at 200 W for 80 min were the most effective for MAE and UAE of the Gac peel samples, respectively. The maximum carotenoid and antioxidant capacity yields of UAE were significantly higher than those of the MAE. The antioxidant capacity of extract obtained by the UAE was also significantly higher that of the conventional extraction using the same ratio of solvent to material. The results showed that both MAE and UAE could be used to reduce the extraction time significantly in comparison with conventional extraction of Gac peel while still obtained good extraction efficiencies. Thus, MAE and UAE are recommended for the improvement of carotenoid and antioxidant capacity extraction from Gac peel.

A green tea extract lowers plasma cholesterol by inhibiting cholesterol synthesis and upregulating the LDL receptor in the cholesterol-fed rabbit.

Green tea extracts enriched in catechins decrease plasma cholesterol in hamsters, mice and rats. The aims of this study were to determine whether a catechin-enriched extract of green tea could lower plasma cholesterol in the cholesterol-fed rabbit and to determine the mechanism of action. Four groups of six New Zealand White rabbits were initially made hypercholesterolaemic by feeding a 0.25% (w/w) cholesterol diet for 2 weeks before the diet was supplemented with a catechin extract from green tea at 0, 0.5, 1 or 2% (w/w) for 4 weeks. Administration of the crude catechin extract from green tea significantly (p<0.05) lowered cholesterol in plasma (-60%), VLDL+IDL (-70%), LDL (-80%), liver (total by -25% and unesterified by -15%) and aorta (-25%) compared to control. There was a significant reduction in the cholesterol synthesis index (-60%) and a significant increase in hepatic LDL receptor activity (+80%) and protein (+70%) but there was no change in the intrinsic capacity to absorb cholesterol from the intestines. These results suggest that green tea catechins lowered plasma, liver and aortic cholesterol in the cholesterol-fed rabbit by lowering cholesterol synthesis and upregulating the hepatic LDL receptor.

A green tea catechin extract upregulates the hepatic low-density lipoprotein receptor in rats.

Green tea extracts have hypocholesterolaemic properties in epidemiological and animal intervention studies. Upregulation of the low-density lipoprotein (LDL) receptor may be one mechanism to explain this as it is the main way cholesterol is removed from the circulation. This study aimed to determine if a green tea extract could upregulate the hepatic LDL receptor in vivo in the rat. A green tea extract (GTE) enriched in its anti-oxidant constituents, the catechins, was fed to rats (n = 6) at concentrations of either 0, 0.5, 1.0 or 2.0% (w/w) mixed in with their normal chow along with 0.25% (w/w) cholesterol for 12 days. Administration of the GTE had no effect on plasma total or LDL cholesterol concentrations but high-density lipoprotein significantly increased (41%; p < 0.05). Interestingly, there was a significant increase in LDL receptor binding activity (2.7-fold) and LDL receptor protein (3.4-fold) in the 2% (w/w) treatment group compared to controls. There were also significant reductions in liver total and unesterified cholesterol (40%). Administration of the GTE significantly reduced cholesterol absorption (24%) but did not affect cholesterol synthesis. These results show that, despite no effect on plasma cholesterol, the GTE upregulated the LDL receptor in vivo. This appears to be via a reduction in liver cholesterol concentration and suggests that the green tea extract was able to increase the efflux of cholesterol from liver cells.

G80A reduced folate carrier SNP influences the absorption and cellular translocation of dietary folate and its association with blood pressure in an elderly population.

The functional consequences of the G80A RFC SNP on the expressed reduced folate carrier protein were evaluated by looking at the relationship between intake of folate, plasma folate and cellular stores of the vitamin. The effect on homocysteine was also examined. Homocysteine is a thiol that is known to be inversely associated with folate, and which is considered to be both thrombo- and athrogenic. At high levels, homocysteine may also interfere with nitric oxide mediated vasodilation, cause oxidative injury to, and proliferation of the vascular endothelium, and alter the elastic properties of the vascular wall, contributing to increased blood pressure. Participants (119; 52 male, 67 female) from a NSW retirement village were assessed. Independent of gender, the assimilation of folate from dietary sources into red cells showed a significant association for GG (r=0.399; p=0.022) and GA (r=0.564; p<0.0001) subjects, but not homozygous recessive (AA) individuals (r=0.223; p=0.236). The same genotype based pattern of significance was shown for the association between dietary folate and plasma folate (GG: r=0.524; p=0.002, GA: r=0.408; p=0.002). No genotype-related pattern of significance was shown for the association between dietary folate and homocysteine. When examined by gender, some differences were apparent; one-way ANOVA showed that genotype influenced diastolic blood pressure in males (p=0.019), while only females showed a significant correlation between dietary folate and blood pressure within specific genotypes (Systolic pressure GA: r=-0.372; p=0.025, carriage of A: r=0.-0.357; p=0.011. Diastolic pressure GA: r=-0.355; p=0.034, carriage of A: r=0.-0.310; p=0.029). The G80A RFC SNP had an impact on the absorption and cellular translocation of dietary folate and its association with blood pressure in an elderly population.

Modulation of cholesterol metabolism by the green tea polyphenol (-)-epigallocatechin gallate in cultured human liver (HepG2) cells.

Epidemiological and animal studies have found that green tea is associated with lower plasma cholesterol. This study aimed to further elucidate how green tea modulates cholesterol metabolism. When HepG2 cells were incubated with the main green tea constituents, the catechins, epigallocatechin gallate (EGCG) was the only catechin to increase LDL receptor binding activity (3-fold) and protein (2.5-fold) above controls. EGCG increased the conversion of sterol regulatory element binding protein-1 (SREBP-1) to its active form (+56%) and lowered the cellular cholesterol concentration (-28%). At 50 microM, EGCG significantly lowered cellular cholesterol synthesis, explaining the reduction in cellular cholesterol. At 200 microM EGCG, cholesterol synthesis was significantly increased even though cellular cholesterol was lower, but there was a significant increase seen in medium cholesterol. This indicates that, at 200 microM, EGCG increases cellular cholesterol efflux. This study provides mechanisms by which green tea modulates cholesterol metabolism and indicates that EGCG might be its active constituent.

The Effects of Resveratrol Supplementation in Overweight and Obese Humans: A Systematic Review of Randomized Trials.

BACKGROUND: Obesity and metabolic syndrome are significant global health issues, with current public health messages predominately focused on altering dietary and physical activity behaviors. Resveratrol is a polyphenol (stilbenoid) commonly found in grapes, and human trials to date have shown conflicting and limited beneficial effects with respect to health. The aim of this study was to determine the effect of resveratrol supplementation on reducing body weight and modifying associated inflammatory markers. METHODS: A systematic review was undertaken following the PRISMA guidelines and using five indexed databases (OVID MEDLINE, Cochrane Library, Web of Science, SCOPUS, and CINAHL). A search strategy was formulated to select randomized, double-blind, placebo-controlled human trials investigating the effects of resveratrol supplementation on obesity or overweight, including body weight, metabolic and inflammatory markers. RESULTS: Five thousand five hundred sixty-nine studies published from 1990 to November 2015 were identified, with only nine papers meeting the inclusion criteria. The studies involved 208 participants (aged 49.2 ± 8.3 years) and utilized a substantial range of resveratrol doses (75-3000 mg/day). Study durations were a minimum of 2 weeks (14-90 days). Seven studies indicated no significant change in body mass index or body weight (P > 0.05), and three studies showed no improvements in fat mass, fat volume, or abdominal fat distribution (P > 0.05). Four studies included measurements of inflammatory markers, with three of these finding resveratrol supplementation to have a significant positive effect (P > 0.05). CONCLUSION: Based on the included studies, there is currently insufficient evidence to support the recommendation of resveratrol supplements in management of obesity. However, there were significant but not entirely consistent anti-inflammatory effects after resveratrol supplementation in overweight and obese individuals.

Effects of maturity on physicochemical properties of Gac fruit (Momordica cochinchinensis Spreng.).

The aril around the seeds of Gac fruit is rich in fatty acids and carotenoids (lycopene and ß-carotene). Understanding how these qualities are affected by fruit maturity at harvest may identify indices for quality assessment. Some physical and chemical properties of Gac fruit were determined for fruit harvested between 8 and 16 weeks after pollination (WAP). Fruit respiration rates and ethylene production rates were assessed after harvest and up to 20 days in storage at 20°C. Fruit harvested at 14 WAP had the highest oil (0.27 ± 0.02 g/g DW), lycopene content (0.45 ± 0.09 mg/g FW), and ß-carotene content (0.33 ± 0.05 mg/g FW) which declined by 16 WAP. External skin color and aril TSS were indicative of oil and carotenoid contents in aril. Skin color, TSS and potentially firmness were good indices of fruit quality. Harvesting less mature fruit at 12 WAP would be practical as the fruit were firmer and more capable of transport; however, quality during postharvest ripening may be limited. Fruits continued to ripen after they were harvested and an ethylene peak in the least mature fruit may reflect a climacteric behavior but this needs further investigation.