Flaxseed (Linum usitatissimum L.) and Its Total Non-digestible Fraction Influence the Expression of Genes Involved in Azoxymethane-induced Colon Cancer in Rats

The influence of flaxseed (Linum usitatissimum L.) and its total non-digestible fraction (TNDF) on the expression of genes involved in azoxymethane (AOM)-induced colon cancer in Sprague Dawley rats was analyzed. The dose used in the animal model was two tablespoons of flaxseed per day, which is the dose recommended for humans. Flaxseed significantly decreased the crypt multiplicity (10.50?±?3.5) compared with the AOM treatment (34.00?±?11.0), which suggests that flaxseed exhibits a preventive effect against colon cancer. Both treatments (flaxseed and TNDF) influence the overexpression of genes involved in cell cycle arrest and mitochondrial apoptosis: p53, p21, bcl-2, bax and caspase-3. Flaxseed induced the expression of p53 and p21, whereas TNDF triggered the p21-independent expression of p53. This finding suggests that both of these treatments induced cell cycle arrest. In addition, TNDF induced mitochondrial apoptosis because the TNDF + AOM group exhibited the expression of caspase-3, decreased bcl-2 expression and increased bax expression. These results suggest that the expression of the analyzed genes is associated with the presence of dietary antioxidants linked to the cell wall of flaxseed.     

Effect of natural fermentation on protein fractions and in vitro protein digestibility of rice

Seaweeds are rich sources of dietary fiber which possess diverse physiological functions in the colon. In the present study, the modulating effect of feeding seatangle (Laminaria japonica), a fiber-rich seaweed, on initiation and postinitiation of rat colon carcinogenesis was investigated. Four groups of animals were exposed to two weekly injections of a chemical carcinogen azoxymethane (AOM). Animals were fed a fiber-free semisynthetic diet or a diet containing 15% seatangle (4.7 g dietary fiber/100 g diet) during initiation, postinitiation or initiation postinitiation phase of carcinogenesis. Study results showed that animals fed the seatangle-containing diet had a significantly decreased number of both aberrant crypts and aberrant crypt foci in the colon. Also, labeling indices and proliferation zones were signi- ficantly reduced in the colon of the rats fed a seatangle diet. All of the seatangle feeding regimens used in this study were effective in reducing aberrant crypt formation indicating anticarcinogenic effects are mediated through both the blocking of initiation and the suppression of cell proliferation in initiated cells. These results suggest that a diet containing seatangle may decrease the risk of developing colon cancer although the precise cellular mechanisms involved in anticarcinogenic effects of seatangle should be further examined.     

Synthesis and Antiproliferative Activity of Thiazolyl-bis-pyrrolo[2,3-b]pyridines and Indolyl-thiazolyl-pyrrolo[2,3-c]pyridines,Nortopsentin Analogues

Two new series of nortopsentin analogues, in which the imidazole ring of the natural product was replaced by thiazole and indole units were both substituted by 7-azaindole moieties or one indole unit was replaced by a 6-azaindole portion, were efficiently synthesized. Compounds belonging to both series inhibited the growth of HCT-116 colorectal cancer cells at low micromolar concentrations, whereas they did not affect the viability of normal-like intestinal cells. A compound of the former series induced apoptosis, evident as externalization of plasma membrane phosphatidylserine (PS), and changes of mitochondrial trans-membrane potential, while blocking the cell cycle in G2/M phase. In contrast, a derivative of the latter series elicited distinct responses in accordance with the dose. Thus, low concentrations (GI₃₀) induced morphological changes characteristic of autophagic death with massive formation of cytoplasmic acid vacuoles without apparent loss of nuclear material, and with arrest of cell cycle at the G1 phase, whereas higher concentrations (GI₇₀) induced apoptosis with arrest of cell cycle at the G1 phase.     

New Capoamycin-Type Antibiotics and Polyene Acids from Marine Streptomyces fradiae PTZ0025

Capoamycin-type antibiotics (2–5) and polyene acids (6, 7) were isolated from marine Streptomyces fradiae strain PTZ0025. Their structures were established by extensive nuclear magnetic resonance (NMR) and high resolution electron spray ionization mass spectroscopy (HRESIMS) analyses and chemical degradation. Compounds 3, 4, 6, 7 were found to be new and named as fradimycins A (3) and B (4), and fradic acids A (6) and B (7). Compounds 3–5 showed in vitro antimicrobial activity against Staphylococcus aureus with a minimal inhibitory concentration (MIC) of 2.0 to 6.0 μg/mL. Interestingly, Compounds 3–5 also significantly inhibited cell growth of colon cancer and glioma with IC₅₀ values ranging from 0.13 to 6.46 μM. Fradimycin B (4), the most active compound, was further determined to arrest cell cycle and induce apoptosis in tumor cells. The results indicated that fradimycin B (4) arrested the cell cycle at the G₀/G₁ phase and induced apoptosis and necrosis in colon cancer and glioma cells. Taken together, the results demonstrated that the marine natural products 3–5, particularly fradimycin B (4), possessed potent antimicrobial and antitumor activities.     

Lipid-Soluble Ginseng Extract Induces Apoptosis and G0/G1 Cell Cycle Arrest in NCI-H460 Human Lung Cancer Cells

This study was performed to elucidate the anticancer mechanism of a lipid-soluble ginseng extract (LSGE) by analyzing induction of apoptosis and arrest of cell cycle progression using the NCI-H460 human lung cancer cell line. Proliferation of NCI-H460 cells was potently inhibited by LSGE in a dose-dependent manner. The cell cycle arrest at the G0/G1 phase in NCI-H460 cells was induced by LSGE. The percentage of G0/G1 phase cells significantly increased, while that of S phase cells decreased after treatment with LSGE. The expression levels of cyclin-dependent kinase2 (CDK2), CDK4, CDK6, cyclin D3 and cyclin E related to G0/G1 cells progression were also altered by LSGE. In addition, LSGE-induced cell death occurred through apoptosis, which was accompanied by increasing the activity of caspases including caspase-8, caspase-9 and caspase-3. Consistent with enhancement of caspase activity, LSGE increased protein levels of cleaved caspase-3, caspase-8, caspase-9, and poly-ADP-ribose polymerase (PARP). These apoptotic effects of LSGE were inhibited by the pan-caspase inhibitor Z-VAD-fmk. These findings indicate that LSGE inhibits NCI-H460 human lung cancer cell growth by cell cycle arrest at the G0/G1 phase and induction of caspase-mediated apoptosis.     

Opuntia humifusa Partitioned Extracts Inhibit the Growth of U87MG Human Glioblastoma Cells

Opuntia humifusa, a member of the Cactaceae family widely distributed in the southern regions of the Korean peninsula, has potential bioactive functions and medicinal benefits. In the present study, we investigated the effect of hexane, ethyl acetate extracts and water partitioned fraction of O. humifusa on proliferation, G1 arrest and apoptosis in U87MG human glioblastoma cells. Glioblastoma cellular proliferation was evaluated using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, and the effects of O. humifusa partitioned extracts on cell cycle and apoptosis were analyzed by flow cytometry. Our results revealed that when U87MG cells were treated with hexane extracts and water partitioned fraction of O. humifusa, the number of viable cells decreased in a concentration-dependent manner. In addition, water partitioned fractions of O. humifusa induced G1 arrest and non-apoptotic cell death as well as significant increases in ROS production in U87MG cells. In conclusion, water partitioned fractions of O. humifusa induce G1 arrest and inhibit U87MG human glioblastoma cell proliferation.     

Effect of Cooking Procedures and Storage on Starch Bioavailability in Common Beans (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.)

Common commercial beans were cooked using two procedures: under pressure (autoclaving) and traditional cooking. Total starch extraction was higher in beans cooked with the traditional procedure (41.69–42.81%) than in the autoclaved samples (37.04–38.16%) and did not change during storage at 4 °C. However, available and total resistant starch levels in vitro were not influenced by the cooking procedure or storage. Retrograded resistant starch content was higher in beans cooked with the traditional process (2.65–2.79%) than in autoclaved beans (1.62–1.94%). The initial in vitro -amylolysis rate in freshly cooked beans was higher in the autoclaved preparation than in the beans cooked by the traditional process, but final hydrolysis indices (90 min) were similar for both samples. None of the bean samples showed statistical differences in -amylolysis behavior (=0.05) after storage at 4 °C for 96 hour.     

Changes in dietary fiber content and its composition as affected by processing of black beans (Phaseolus vulgaris,Tamazulapa variety)

This paper presents the effect that the traditional cooking process of black beans (Phaseolus vulgaris, Tamazulapa variety) has on the quantity and composition of soluble (SDF) and insoluble (IDF) dietary fiber of beans, as well as on its protein digestibility and protein quality. There was an increase of IDF from 18.1% in cooked beans to 22.4% in fried beans, and a decrease in SDF from 8.4% to 6.6%, respectively. Starch content decreased from 34.5% to 31.3%. No change was found in lignin. The xylose content was higher in IDF than in SDF and decreased to some extent from cooked to fried beans. Arabinose content was similar in IDF and SDF with no change caused by processing. The fraction containing glucose, mannose and galactose in IDF was higher than in SDF, the content increasing in IDF and decreasing in SDF, with processing. Protein content in IDF was higher than in SDF, with no major change when processing. About 29.5% of the total protein of beans was bound in DF. Protein digestibility and protein quality decreased from cooked to fried beans and was positively related to IDF.     

Antitumor Effects of a Sesquiterpene Derivative from Marine Sponge in Human Breast Cancer Cells

In this study, the anti-proliferative effect of ilimaquinone, a sesquiterpene derivative from the marine sponge, in breast cancer cells was investigated. Ilimaquinone inhibited the proliferation of MCF-7 and MDA-MB-231 breast cancer cells with IC₅₀ values of 10.6 μM and 13.5 μM, respectively. Non-tumorigenic human breast epithelial cells were less sensitive to ilimaquinone than breast cancer cells. Flow cytometric and Western blot analysis showed that ilimaquinone induced S-phase arrest by modulating the expression of p-CDC-2 and p21. Ilimaquinone induces apoptosis, which is accompanied by multiple biological biomarkers, including the downregulation of Akt, ERK, and Bax, upregulation of p38, loss of mitochondrial membrane potential, increased reactive oxygen species generation, and induced autophagy. Collectively, these findings suggest that ilimaquinone causes cell cycle arrest as well as induces apoptosis and autophagy in breast cancer cells.     

Quality attributes of three new improved lines of Nigerian lima beans (Phaseolus lunatus L. Walp.)

Three promising new improved lines of lima beans (Tpl 1^B, Tpl 7^A and Tpl 175^A) were evaluated for physicochemical properties and cooking quality. The beans varied in seed dimensions and weights with Tpl 1^B and Tpl 7^A having smaller seed volume than Tpl 175^A. Seed coat percentages, leached solids and swelling capacities were within a range of 10.2–19.6% (w/w), 0.44–0.92 g/100 g and 94.0–121.0 g/100 g dry bean, respectively. Cooking times varied between 62 and 81 min without soaking and were reduced by about 34%following a presoaking treatment in water for 12 h at room temperature (28 ± 1 °C). Small seeds absorbed higher amounts of water during soaking and required more cooking time than larger seeds. No significant (p>0.05) difference in cooked texture was found between unsoaked beans cooked for 50 min and soaked beans cooked for 30 min,suggesting that cooking times and cooked texture for all lines were improved through soaking.     

Evaluation of selected food characteristics of three advanced lines of Nigerian soybean (Glycine max (L.) Merr.)

The seeds of three promising advanced lines of soybeans (TGx 923-2^EN, TGx 1019-2^EN and TGx 1497-1^D) which were part of a larger collection evaluated in agronomic field trials in Nigeria were selected for characterization of physicochemical properties, chemical composition, water absorption, cooking time and cooked texture as a function of soaking and cooking. Seed density, leached solids, swelling capacity and seed coat percentage were within a range of 1.15 to 1.26 g per ml, 1.00 to 1.26 g per 100 g, 80.25 to 84.35 g per 100 g and 6.6 to 10.1% w/w of dry beans, respectively. The total polyphenol content of the cream colored beans was similar (0.75 to 0.76 mg/g) but higher than the amount (0.60 mg/g) found in the white beans. Cooking times varied between 71 and 96 min without soaking and were reduced by about 32.0% following a presoaking treatment in water for 12 hours at room temperature (28±1°C). Small seeds absorbed higher amounts of water during soaking and required less cooking time than larger seeds. Unsoaked beans required 40 min of cooking to achieve the same degree of cooked texture as the soaked beans cooked for 20 min, suggesting that cooking times and cooked texture for all lines were improved through soaking.     

Purified Brominated Indole Derivatives from Dicathais orbita Induce Apoptosis and Cell Cycle Arrest in Colorectal Cancer Cell Lines

Dicathais orbita is a large Australian marine gastropod known to produce bioactive compounds with anticancer properties. In this research, we used bioassay guided fractionation from the egg mass extract of D. orbita using flash column chromatography and identified fractions containing tyrindoleninone and 6-bromoisatin as the most active against colon cancer cells HT29 and Caco-2. Liquid chromatography coupled with mass spectrometry (LCMS) and ¹H NMR were used to characterize the purity and chemical composition of the isolated compounds. An MTT assay was used to determine effects on cell viability. Necrosis and apoptosis induction using caspase/LDH assay and flow cytometry (PI/Annexin-V) and cell cycle analysis were also investigated. Our results show that semi-purified 6-bromoisatin had the highest anti-cancer activity by inhibiting cell viability (IC₅₀ = ~100 µM) and increasing caspase 3/7 activity in both of the cell lines at low concentration. The fraction containing 6-bromoisatin induced 77.6% apoptosis and arrested 25.7% of the cells in G2/M phase of cell cycle in HT29 cells. Tyrindoleninone was less potent but significantly decreased the viability of HT29 cells at IC₅₀ = 390 µM and induced apoptosis at 195 µM by increasing caspase 3/7 activity in these cells. This research will facilitate the development of these molluscan natural products as novel complementary medicines for colorectal cancer.     

Effects of soaking,cooking and fermentation on composition,in-vitro starch digestibility and nutritive value of common beans

A common bean variety, grown in Burundi, was either fermented, soaked and/or cooked, and then assessed for nutrient composition, in-vitro starch digestibility and protein nutritive value. A decrease in ash, most minerals, vitamins, and some essential amino acids was noted for soaked, cooked and soaked-cooked beans. Compared to untreated beans, soaking decreased soluble sugar (9.8 percent) but increased starch (7.3 percent) and soluble fiber (16.9 percent). In cooked beans, an increase in soluble sugar (1.5 percent), and a decrease in thiamine (81.7 percent), starch (24.6 percent) and soluble fiber (16.6 percent) and nitrogen (2.9 percent) contents were observed. Crude fiber (6.9 percent) and starch (10.0 percent) increased while fat (17.6 percent), fatty acids (linoleic: 10.7 percent; linolenic: 14.3 percent) and soluble sugars (25.4 percent) and nitrogen (14.4 percent) decreased in soaked-cooked beans. Fermentation increased potassium (11.6 percent), soluble fiber (18.9 percent), and some amino acids but decreased fatty acids (linoleic: 13.5 percent; linolenic: 19.9 percent), soluble sugar (75.2 percent) and vitamin (riboflavin: 41.0 percent; niacin: 24.5 percent) contents in common beans. However, the in-vitro starch digestibility was greatly improved (12.3 percent) by cooking while it decreased in soaked beans (29.2 percent). Soaking-cooking and fermentation did not have any significant effect on the digestibility of common bean starch. Finally, among the five treatments applied to common beans, only fermentation showed a significant improvement (8.3 percent) on the protein nutritive value of this legume.     

Fibre digestibility and its relationships with chemical and morphological traits in thirty‐two sugarcane varieties

The objectives of this study were to determine the range of neutral detergent fibre (NDF) digestibility across sugarcane varieties and predict dry‐matter (DM) and NDF digestibility based on morphological and chemical composition. Whole plants of thirty‐two commercial varieties were collected after 12 months regrowth (second cut) and evaluated for morphological traits, chemical composition, in situ digestibility of NDF (NDFD) and of DM (DMD), in vitro NDFD (IVNDFD) and gas production dynamics from isolated NDF. The content of NDF ranged from 404 to 547 g kg^?1 DM, DMD was between 593 and 739 g kg^?1 and NDFD varied from 270 to 363 g kg^?1. Cluster and discriminant analyses for NDFD agglomerated sugarcane genotypes into three groups (high, medium and low NDFD). The contents of NDF and lignin were the best univariate predictors of sugarcane DMD and NDFD respectively. Therefore, associating NDF and lignin improved the accuracy of DMD prediction. Associating lignin concentration with the number of dry leaves and stalk length significantly improved the goodness of fit of NDFD prediction. Contrary to expectations, silicon concentration was not related to NDFD. The principal component analysis patterned the content of neutral detergent solubles, NDFD and variables related to DM yield in different components. Hence, these characteristics would coexist in the same variety. Based on Lucas test and kinetics of gas production from NDF, sugarcane potentially digestible NDF (pdNDF) can be considered almost uniform (pdNDF digestibility = 677 g kg^?1). In conclusion, associating chemical composition and morphological traits can successfully improve the screening of sugarcane genotypes for animal nutrition.     

Induction of Apoptosis, G0/G1 Phase Arrest and Microtubule Disassembly in K562 Leukemia Cells by Mere15, a Novel Polypeptide from Meretrix meretrix Linnaeus

Mere15 is a novel polypeptide from Meretrix meretrix Linnaeus with cytotoxicity in solid cancer cells. In this study, we investigated its activity on human K562 chronic myelogenous leukemia cells. Mere15 inhibited the growth of K562 cells with IC₅₀ values of 38.2 μg/mL. Mere15 also caused concentration dependent induction of apoptosis, with overproduction of reactive oxygen species and loss of mitochondrial membrane potential. Moreover, Mere15 arrested cell cycle progression at G₀/G₁ phase of K562 cells in a concentration dependent manner. In addition, Mere15 caused the disassembly of the microtubule cytoskeleton in K562 cells and inhibited the polymerization of tubulin in a cell free system via interaction with tubulin. We concluded that Mere15 was cytotoxic to K562 leukemia cells and the cytotoxicity was related to the apoptosis induction, cell cycle arrest and microtubule disassembly. These results implied that Merer15 was a broad spectrum anticancer polypeptide, not only cytotoxic to various solid cancer cells but also to the chronic myelogenous leukemia cells. Mere15 may have therapeutic potential for the treatment of leukemia.     

The effects of forage preservation method and proportion of concentrate on digestion of cell wall carbohydrates and rumen digesta pool size in cattle

Six bulls with ruminal and duodenal cannulae were used in a 6 × 6 Latin square design with a 2 × 3 factorial arrangement of treatments. Barn-dried grass (G) and direct-cut silage (S) from the same sward were fed together with 250 (L), 500 (M) and 750 (H) g kg^-1 total dry matter (DM) of a barley-based concentrate (barley 875 and rape-seed meal 125 g kg^-1) at the level of 80 g DM kg^-1 live weight^0·75. Rumen and total digestibility of cell wall constituents were measured by a double marker and total collection method. Rumen pool sizes of dietary constituents were estimated by emptying the rumen. Particle-associated enzyme activities were measured from rumen particulate material and feed particles incubated in nylon bags in the rumen. Neutral detergent fibre (NDF) and hemicellulose digestibility were higher (P<0·05) for G diets than for S diets while no differences were observed between the forages in acid detergent fibre (ADF) and cellulose digestibility. Both rumen and total digestibility of cell wall constituents decreased with increasing level of concentrate. The proportion of total cell wall digestion in the rumen was unaffected by the forage preservation method and the proportion of concentrate. NDF, and especially cellulose digestibility, declined quadratically with increasing level of concentrate. Dietary effects on particle-associated carboxymethylcellulase and xylanase activity were consistent with those observed in cell wall digestion. There were no differences between the forages in rumen pool size of total ingesta or any dietary constituent. Rumen pool size of total ingesta decreased with increasing level of concentrate. On the other hand, DM content of ingesta increased with the level of concentrate, while the pool size of DM, NDF and ADF declined quadratically reaching a minimum on M level of concentrate. The differences in rumen NDF pool size were mainly in the digestible fraction. Calculation of digestion kinetic parameters showed that both the rate of passage and especially the rate of digestion were markedly depressed in animals fed on the highest level of concentrate. The results suggest that a reduced fate of digestion of NDF with high concentrate diets can be partially compensated for by an increased NDF retention time in the rumen of animals fed at a restricted level of feeding.     

Effects of altitude above sea level on the cooking time and nutritional value of common beans

The present study was conducted with the objective to determine the effects of altitude above sea level, on the cooking time and nutritional value of common black beans (Phaseolus vulgaris). Three 100 g samples of theOstua variety were cooked at 8 individual locations, ranging in altitude from 0 to 2256 meters, in Guatemala, to establish water uptake and cooking time. The cooked samples were separated into cooked beans and cooking broth for chemical analysis. This included moisture, protein, lysine, tannins, total and enzyme susceptible starch, and fiber fractionation. The cooking liquor was analyzed for total solids, moisture, protein, ash and K. A 1200 g sample was cooked for the cooking time established previously, for biological testing of nutritional value,which included Net Protein Ratio (NPR), Protein Effciency Ratio (PER), and protein digestibility. Altitude influenced cooking time which increased from 78 min at 0 m, to 264 min at 2256 m. Final moisture content in the cooked bean was similar at all altitudes and there was a tendency to yield smaller amounts of solids in the cooking broth at higher altitudes. The increase in cooking time was significant. Bean water uptake at all times was significantly slower and smaller at ambient T, as compared to water uptake at boiling T, at all altitudes. Protein and lysine content were not affected by altitude, however, tannin and catechin were lower in cooked samples, as compared to the raw material. Altitude did not affect the content of these substances. Total starch and total sugars were higher in the raw sample, as compared to the cooked samples, but there was no effect of altitude. Enzyme susceptible starch (ESS) was lower in the raw sample as compared to the cooked samples, which contained similar amounts with respect to altitude. No change was observed in fiber fractions of the cooked beans. Likewise, the composition of the cooking broth was very similar between cooking locations. There was a small tendency to a lower protein quality, with respect to altitude, the effect of which was more obvious in the apparent protein digestibility values. Undercooking or overcooking at one location influenced protein quality values. The extended cooking time of beans at high altitudes, has important economic and environmental implications, since significant amounts of wood have to be used.     

The Marine Fungal Metabolite,Dicitrinone B,Induces A375 Cell Apoptosis through the ROS-Related Caspase Pathway

Dicitrinone B, a rare carbon-bridged citrinin dimer, was isolated from the marine-derived fungus, Penicillium citrinum. It was reported to have antitumor effects on tumor cells previously; however, the details of the mechanism remain unclear. In this study, we found that dicitrinone B inhibited the proliferation of multiple tumor types. Among them, the human malignant melanoma cell, A375, was confirmed to be the most sensitive. Morphologic evaluation, cell cycle arrest and apoptosis rate analysis results showed that dicitrinone B significantly induced A375 cell apoptosis. Subsequent observation of reactive oxygen species (ROS) accumulation and mitochondrial membrane potential (MMP) reduction revealed that the apoptosis induced by dicitrinone B may be triggered by over-producing ROS. Further studies indicated that the apoptosis was associated with both intrinsic and extrinsic apoptosis pathways under the regulation of Bcl-2 family proteins. Caspase-9, caspase-8 and caspase-3 were activated during the process, leading to PARP cleavage. The pan-caspase inhibitor, Z-VAD-FMK, could reverse dicitrinone B-induced apoptosis, suggesting that it is a caspase-dependent pathway. Our data for the first time showed that dicitrinone B inhibits the proliferation of tumor cells by inducing cell apoptosis. Moreover, compared with the first-line chemotherapy drug, 5-fluorouracil (5-Fu), dicitrinone B showed much more potent anticancer efficacy, suggesting that it might serve as a potential antitumor agent.