Abstract: The objective of this study was to investigate the uptake of deltamethrin, an insecticide, by Daphnia magna neonates by SIMS and to compare these findings with results based on established toxicity tests. Young daphnids (aged <24h) were exposed to 0, 50 and 200mugL(-1) (ppb) deltamethrin. Mobile, immobile and dead animals were enumerated after 24 and 48h following OECD 202 [OECD 202, 2004. Daphnia sp., acute immobilisation test, guideline for testing of chemicals] guidelines. The animals were embedded in epoxy resin, cut into semi-thin sections (500nm) and placed on silicon supporters. NanoSIMS 50 (Cameca) images were made from tissues of the intestine for carbon, nitrogen (measured as CN), phosphorus and bromine. To distinguish between relative concentrations of bromine in the guts from different exposure concentrations of deltamethrin, a carbon normalization method was carried out. Both deltamethrin concentrations and time showed a significant effect on immobilization and mortality of the daphnids (P<0.0001). Bromine from deltamethrin could be visualized by NanoSIMS in all exposed gut tissues (gut wall, microvilli layer, perithropic membrane). Highest deltamethrin concentrations following (12)C normalization were found in animals exposed to 200mugL(-1) deltamethrin, followed by 50mugL(-1) and the control. NanoSIMS 50 was successfully used as a supplemental technique for elucidating the relation between the uptake and localization of deltamethrin and its toxicity to D. magna. These results highlight the potential usefulness of NanoSIMS to detect marker elements of xenobiotic compounds within exposed organisms, to compare relative exposure concentrations, and to locate these compounds at their original tissue location.
Abstract: The relative contribution of the small intestine to absorption and microbial metabolism of ingested isoflavonoids (IFN) was investigated in swine with canulae in distal ilea to facilitate collection of chyme (canula open). Weaned swine were fed a single meal containing ground roasted soybean and corn with canulae open followed by a second test soy diet at 48 h with canulae closed to allow passage of chyme into the large intestine. All remaining feedings were soy-free (corn-casein diet). Ileal effluent and urine were collected for 16 and 48 h, respectively, and analyzed for IFN and microbial metabolites of IFN. IFN in ileal effluent were present entirely as aglycones. IFN equivalents excreted for 24 h after ingesting the soy diet were not significantly different when canulae were open or closed. Urinary IFN aglycone equivalents on day 2 were similar to those on day 1 when canulae remained closed, but less than 10% of that on day 1 when canulae were open for 16 h postfeeding. Urinary concentrations of dihydrodaidzein, dihydrogenistein, O-desmethylangolensin, and equol exceeded IFN aglycone equivalents. These findings suggest extensive preabsorptive conversion of IFN glucosides to aglycones in the small intestine and relatively efficient microbial metabolism of IFN in weaned swine.
Abstract: Decreased Mg intake and low Mg status have been associated with a number of major health concerns such as xD;diabetes mellitus type II, coronary heart disease, and osteoporosis. While information on Mg intake is available, relatively xD;little is known on dietary factors influencing Mg bioavailability. While it is established that Mg absorption is based on a xD;combination of a non-saturable and a saturable pathway, the nature of especially the latter mechanism is not well xD;understood. Recently, stable isotopes have improved techniques available for the determination of Mg absorption from xD;single test meals or supplements. Some inorganic Mg forms such as MgO seem of limited solubility in the intestine, xD;suggesting low bioavailability. Recent studies have further added evidence that some commonly consumed dietary xD;compounds, such as phytate and oxalate, can inhibit Mg absorption, presumably via complexation, preventing absorption xD;from the small intestine. Phytate for example has been shown to decrease Mg absorption by up to 60%, in a dose xD;dependent manner. On the other hand, fermentable dietary fibre, such as fructo-oligosaccharides, have been demonstrated xD;to increase Mg absorption in humans by 10-25%, even though the underlying mechanisms remain to be elucidated. Future xD;studies to investigate factors impacting Mg absorption are warranted.
Abstract: AIMS: NanoSIMS (secondary ion mass spectrometry) is a powerful technique for mapping the elemental composition of a variety of small-scale samples (e.g. in Material Research, Cosmochemistry and Geology). However, its analytical features are making it also valuable to address biological questions. We demonstrate the ability of the NanoSIMS 50 to map elements at subcellular lateral resolution (approx. 50 nm) within cyanobacteria (Anabaena sp. and Cylindrospermum alatosporum) and its feasibility to investigate the uptake of bromine-containing substances (NaBr and deltamethrin). METHODS AND RESULTS: Elemental maps of O, N, P and S were obtained from semi-thin sections of different cell types (chemically fixed and resin-embedded heterocysts, akinetes and vegetative cells). NanoSIMS enabled the detection of various characteristic cell sub-structures and inclusions. A homogenous bromine distribution was detected following NaBr and deltamethrin exposure, at Br-concentrations of 0.05, 0.5 (NaBr) and 0.0025 mmol l(-1) (deltamethrin). CONCLUSIONS: NanoSIMS allowed study of the mapping of common elements in cyanobacterial cells and the uptake of NaBr and deltamethrin. SIGNIFICANCE AND IMPACT OF THE STUDY: These results highlight the potential usefulness of NanoSIMS analysis for tracking elements within cell structures at the nanoscale and the ability to detect marker elements of xenobiotic compounds within exposed organisms.
Abstract: The role of carotenoids in human nutrition has gained increased interest, especially due to their associated xD;health-beneficial effects for a number of chronic diseases, including certain types of cancer and cardiovascular disease. xD;Whereas data is available on the intake and presence of carotenoids in foods, limited information exists on factors xD;influencing their bioavailability, especially for the non-provitamin A carotenoids. However, carotenoid absorption xD;strongly depends on a number of factors which are not entirely understood. These include mainly the release of xD;carotenoids from the food matrix, their incorporation into mixed bile micelles, the transfer of carotenoids from micelles to xD;the mucosa for passive or facilitated absorption (via SR-BI proteins), and the sequestration into chylomicrons. Thus, xD;dietary compounds influencing carotenoid micelle incorporation, e.g. the amount fat present in an ingested meal or xD;components competing for uptake, such as phytosterols and other carotenoids, can have a considerable impact on xD;carotenoid bioavailability. However, the effect of many dietary factors, including dietary fiber, type of fat, or minerals on xD;carotenoid absorption is not well understood. In addition, bioavailability also depends on the carotenoid structure; in xD;general, polar carotenoids are preferably incorporated into mixed micelles and tend to be of higher bioavailability, as may xD;be the case for free vs. esterified xanthophylls, and cis-isomers vs. their trans-form, due to apparent shorter chain-lengths. xD;Whereas their importance as part of a healthy diet warrants an improved understanding of carotenoids, their dietary fate xD;following ingestion, including their stability, efficiency of micelle-incorporation, and pathway of absorption are still xD;marginally understood.
Abstract: Angus-cross steers (n = 165; 295 +/- 16 kg of BW) were used evaluate the effect of low vitamin A diets with high-moisture corn (HMC) or dry corn (DC) on marbling and fatty acid composition. Steers were allotted to 24 pens (7 steers/pen), such that each pen had the same average initial BW. Treatments were randomly allotted to the pens. The experiment had a completely randomized design, with a 2 x 2 factorial arrangement of treatments: low vitamin A (Lo, no supplemental vitamin A) and HMC (LoHMC); LoDC; high vitamin A (Hi, supplemented with 2,200 IU of vitamin A/kg of DM) and HMC (HiHMC); and HiDC. Diets contained 76% corn, 10% corn silage, 11% protein supplement, and 3% soybean oil (DM basis). Samples of feed ingredients were collected for carotenoid analysis. Blood samples were collected for serum retinol determination. Steers were slaughtered after 145 d on feed. Carcass characteristics and LM composition were determined. Samples from the s.c. fat depot were analyzed for fatty acid composition. High-moisture corn had a greater vitamin A content, based on its carotenoid content, than DC (614 vs. 366 IU/kg of DM, P < 0.01). No vitamin A x corn type interactions were detected for feedlot performance, carcass characteristics, or serum, s.c. fat, or liver retinol concentration. Average daily gain, DMI, and G:F were not affected by vitamin A (P > 0.05). Marbling score and USDA quality grade were greater (P < 0.05) in Lo vs. Hi steers. Hot carcass weight, backfat, and yield grade were not affected by the treatments (P > 0.05). Vitamin A and corn type did not affect LM composition (DM, ash, CP, or ether-extractable fat, P > 0.05). Vitamin A supplementation increased (P < 0.06) serum retinol on d 112 and 145 and increased (P < 0.01) liver retinol at slaughter (Lo = 38.7 vs. Hi = 102.9 mug/g). The s.c. fat retinol concentrations were less (P < 0.01) for Lo (0.8 mug/g) than for Hi (1.4 mug/g) at slaughter. Cell diameter of adipocytes in the i.m. depot was not affected by dietary vitamin A (P > 0.05). A vitamin A x corn type interaction was observed (P < 0.05) for the s.c. fat cellularity. Feeding HMC increased the number of cells per square millimeter when Lo diets were fed (LoHMC = 128 vs. LoDC = 100 cells/mm(2), P < 0.05), but not when Hi diets were fed (HiHMC = 109 vs. HiDC = 111 cells/mm(2), P > 0.05). The CLA content of adipose tissue was not affected by the treatments. Regardless of the corn type used, feeding low vitamin A diets for 145 d to Angus-cross steers increased marbling and quality grade without affecting yield grade, animal health, or performance.
Abstract: Tomato sauces were produced from unique tomato varieties to study carotenoid absorption in humans. Tangerine tomatoes, high in cis-lycopene, especially prolycopene (7Z,9Z,7'Z,9'Z), and high-beta-carotene tomatoes as an alternative dietary source of beta-carotene were grown and processed. Sauces were served after 2 week washout periods and overnight fasting for breakfast to healthy subjects (n = 12, 6M/6F) in a randomized crossover design. The serving size was 150 g (containing 15 g of corn oil), tangerine sauce containing 13 mg of lycopene (97.0% as cis-isomers) and high-beta-carotene sauce containing 17 mg of total beta-carotene (1.6% as the 9-cis-isomer) and 4 mg of lycopene. Blood samples were collected 0, 2, 3, 4, 5, 6, 8, and 9.5 h following test meal consumption and carotenoids determined in the plasma triacylglycerol-rich lipoprotein fraction by HPLC-electrochemical detection. Baseline-corrected areas under the concentration vs time curves (AUC) were used as a measure of absorption. AUC0-9.5h values for total lycopene in the tangerine sauce group were 870 +/- 187 (nmol.h)/L (mean +/- SEM) with >99% as cis-isomers (59% as the tetra-cis-isomer). The AUC0-9.5h values for total beta-carotene and lycopene after consumption of the high-beta-carotene sauce were 304 +/- 54 (4% as 9-cis-carotene) and 118 +/- 24 (nmol.h)/L, respectively. Lycopene dose-adjusted triacylglycerol-rich lipoprotein AUC responses in the tangerine sauce group were relatively high when compared to those in the literature and the high-beta-carotene group. The results support the hypothesis that lycopene cis-isomers are highly bioavailable and suggest that special tomato varieties can be utilized to increase both the intake and bioavailability of health-beneficial carotenoids.
Abstract: Phytosterols have been shown to reduce cholesterol absorption in humans. Supplementing phytosterols in fat-free formulations, however, has yielded controversial results. In the present study, we investigated the effect of supplementing test meals with different fat-free phytosterol products on cholesterol incorporation into mixed micelles during simulated digestion and accumulation of micellar cholesterol by Caco-2 cells: control orange juice (OJ), orange juice supplemented with either multivitamin/multimineral tablets (MVT), multivitamin/multimineral tablets containing phytosterols (MVT+P), and phytosterol powder (PP). These combinations were added to Ensure-based test meals and spiked with cholesterol of natural isotopic composition or 13C2-cholesterol to differentiate external from endogenous cholesterol. After simulated gastric/small intestinal digestion, micelle fractions were analyzed for cholesterol enzymatically (n = 6-20/product) and by high-performance liquid chromatography-tandem mass spectrometry (n = 12/product) and added to Caco-2 cells to determine the accumulation of 13C2-cholesterol (n = 10-24/product). As compared to OJ, PP and MVT+P significantly decreased cholesterol micellarization (determined enzymatically) by 70 +/- 39 (mean +/- SD) and 70 +/- 39%, respectively (P < 0.001, Bonferroni). The stable isotope experiments revealed that both PP and MVT+P reduced cholesterol micellarization [by 25 +/- 12 (P = 0.055) and 21 +/- 8% (P = 0.020), respectively, Fisher's protected LSD test] and Caco-2 cell accumulation (by 28 +/- 8 and 10 +/- 8%, respectively; P < 0.010, Bonferroni). OJ+P did not inhibit micellarization or accumulation of cholesterol by Caco-2 cells. This study shows that fat-free phytosterol-containing products can significantly inhibit cholesterol micellarization and Caco-2 cell bioaccessibility, albeit to different extents depending on individual formulations. This is most likely explained by inhibition of cholesterol micellarization.
Abstract: BACKGROUND: Although soy isoflavonoids have a number of health-promoting benefits, information concerning the sites of their absorption and metabolism in humans remains limited. Isoflavonoid absorption from the gut requires deconjugation of glucosides to aglycones. OBJECTIVE: The objective was to investigate the role of the small intestine in isoflavonoid absorption and metabolism in humans. DESIGN: Human subjects with fully functional gastrointestinal tracts (n = 6) and ileostomy subjects (n = 6) were fed a single soy meal containing 64.8 mg isoflavonoid aglycone equivalents (95% as glucosides). Metabolism of isoflavonoids in the upper gastrointestinal tract was examined by analyzing ileal effluent from ileostomy subjects, and absorption was assessed indirectly by quantifying isoflavonoids and several metabolites in 24-h urine pools. RESULTS: Chyme contained 36.7% of ingested isoflavonoid aglycone equivalents, primarily (95.8%) as aglycones. Qualitative profiles (x +/- SEM) of isoflavonoid excretion in urine (daidzein > glycitein > genistein) and the quantity of isoflavonoid equivalents were not significantly different between the control (18.4 +/- 2.2 mg) and ileostomy (13.5 +/- 3.2 mg) subjects. Dihydrodaidzein was present in the urine of all subjects, although the amount excreted by ileostomy subjects was less than that excreted by the control subjects. The percentage of producers and mean quantities of dihydrogenistein, equol, and O-desmethylangolensin in the urine of ileostomy subjects also were lower than those of control subjects. CONCLUSIONS: Ileostomy subjects efficiently deglycosylate isoflavonoid glucosides in the small intestine and appear to absorb aglycones with an efficiency comparable with that of control subjects. However, the production of microbial metabolites of isoflavonoids is limited in ileostomy subjects.
Abstract: A simple, sensitive and selective liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometric method (LC/APCI-MS/MS) was developed and applied to quantitative determination of uptake of cholesterol by Caco-2 human intestine cells. Caco-2 cells were cultured in medium containing cholesterol-3,4-13C2 and phytosterols from nutritional supplements after in vitro digestion. Cellular cholesterol (cholesterol-3,4-13C2) and endogenous cholesterol were extracted using methanol/chloroform (1:2, v/v) and directly analyzed using LC/APCI-MS/MS with selected reaction monitoring (SRM), using cholesterol-2,2,3,4,4,6-d6 as an internal standard. Detection and quantification limits were 2.2 and 7.2 pmol, respectively. This method provides an effective tool for rapid determination of cholesterol uptake by cells with increased selectivity and sensitivity in comparison to previously reported LC/APCI-MS analysis using selected ion monitoring (SIM).
Abstract: Dietary lipids are hypothesized to bean important factor for carotenoid bioavailability. However, most carotenoid-rich fruits and vegetables are low in lipids. The objective of this study was to assess whether the addition of avocado fruit as a lipid source enhances carotenoid absorption in humans. Healthy subjects (n = 11/study) were recruited for 2 crossover, postprandial studies. The effect of avocado addition (150 g) to salsa on lycopene and beta-carotene absorption was examined in Study 1, and the absorption of lutein, alpha-carotene, and beta-carotene from salad in Study 2. Furthermore, the effects of avocado dose (75 vs. 150 g containing 12 vs. 24 g lipid, respectively) and of lipid source (avocado fruit vs. avocado oil) on carotenoid absorption were examined in Study 2. Intact carotenoids were quantified in the plasma triacylglycerol-rich lipoprotein (TRL) fraction during the 9.5 h after consumption of the test meal and expressed as baseline-corrected area under the concentration-vs.-time curve (AUC). The addition of avocado to salsa enhanced lycopene and beta-carotene absorption (P < 0.003), resulting in 4.4 and 2.6 times the mean AUC after intake of avocado-free salsa, respectively. In Study 2, supplementing 150 g avocado or 24 g avocado oil to salad similarly enhanced alpha-carotene, beta-carotene, and lutein absorption (P < 0.01), resulting in 7.2, 15.3, and 5.1 times the mean AUC after intake of avocado-free salad, respectively (150 g avocado). Neither the avocado dose nor the lipid source affected carotenoid absorption. In conclusion, adding avocado fruit can significantly enhance carotenoid absorption from salad and salsa, which is attributed primarily to the lipids present in avocado. xD;
Abstract: Magnesium is bound as the central atom of the porphyrin ring of the green plant pigments chlorophyll a and b. It has been suggested that chlorophyll-bound magnesium may play an important role in magnesium nutrition because when iron is similarly bound in the porphyrin ring of heme, it is absorbed to a greater extent than non-heme iron. We have analyzed 22 frequently consumed fruits and vegetables for the chlorophyll content by high-pressure liquid chromatography and for magnesium with atomic absorption spectroscopy. Chlorophyll concentrations ranged from 6 mug/g (grape) to 790 mug/g (spinach) (median 63 mug/g). Magnesium concentrations ranged from 48 mug/g (grape) to 849 mug/g (spinach) (median 122 mug/g). In the green leafy vegetables, such as spinach and lettuce, chlorophyll-bound magnesium represented 2.5% to 10.5% of total magnesium whereas other common green vegetables, pulses and fruits contained <1% chlorophyll-bound magnesium. The chlorophyll content of spinach was further decreased by about 35% on thawing frozen spinach or on chopping fresh spinach, and this degradation increased to about 50% after boiling and steaming. Based on the present results and published food consumption data, we estimate that chlorophyll-bound magnesium represents a very low fraction of total magnesium intake in industrialized countries, less than 1% in the case for data obtained from Switzerland. Thus, chlorophyll-bound magnesium is of little relevance to magnesium nutrition.
Abstract: Background: Phytic acid has been reported to impair the absorption of minerals and trace elements such as calcium, zinc, and iron in humans. However, limited information is available on the effect of phytic acid on magnesium absorption. Objective: The objective was to evaluate the effect of phytic acid on fractional apparent magnesium absorption in humans. Design: Two stable-isotope studies were performed with 8-9 healthy adults per study. Test meals were based on 200 g phytic acid-free wheat bread; test meals with and without added phytic acid were served on days I and 3 according to a crossover design. Phytic acid was added in amounts similar to those naturally present in whole-meal (1.49 mmol) and in brown bread (0.75 mmol). Each test meal was labeled with 0.7 mmol Mg-25 or 1.1 mmol Mg-26. The total magnesium content was standardized to 3.6 mmol in all test meals. Apparent magnesium absorption was based on fecal monitoring. Results: The addition of phytic acid lowered fractional apparent magnesium absorption from 32.5 +/- 6.9% (no added phytic acid) to 13.0 +/- 6.9% (1.49 mmol added phytic acid; P < 0.0005) and from 32.2 +/- 12.0% (no added phytic acid) to 24.0 +/- 12.9% (0.75 mmol added phytic acid; P < 0.01). The inhibiting effect of phytic acid was dose dependent (P < 0.005). Conclusion: The results show that fractional magnesium absorption from white-wheat bread is significantly impaired by the addition of phytic acid, in a dose-dependent manner, at amounts similar to those naturally present in whole-meal and brown bread.
Abstract: The aim of the present study was to evaluate Mg absorption from a test meal served with an oxalate-rich vegetable, spinach, as compared with a test meal served with a vegetable with a low oxalate content, kale. Mg absorption was measured by a stable-isotope technique based on extrinsic labelling of the test meals and faecal monitoring of the excreted isotope labels. Nine healthy adults participated in the study. The test meals were based on 100 g phytate-free white bread, served with 300 g spinach (6.6 mmol oxalate; 0.7 mmol Mg-25 label added, 5.0 mmol total Mg) or 300 g kale (0.1 mmol oxalate; 1.2 mmol Mg-26 label added, 4.8 mmol total Mg). The test meals were served on days 1 and 3, at breakfast and lunch, using a cross-over design. The results from the present study demonstrated that apparent Mg absorption was significantly lower from the meal served with spinach (26.7 (SD 10.4) %) than the meal served with kale (36.5 (SD 11.8) %) (P=0.01). However, the lower fractional apparent Mg absorption from the test meal served with spinach can be assumed to be, at least partly, counterbalanced by the higher native Mg content of spinach as compared with kale. Although based on indirect evidence, i.e. not based on an evaluation of added (or removed) oxalic acid, the difference in Mg absorption observed in the present study is attributed to the difference in oxalic acid content between the two vegetables.
Abstract: Chlorophyll analysis at high precision and accuracy is limited by the lack of suitable, commercially available internal standards for HPLC analysis. Here, the commercially available dye zinc-phthalocyanine is presented as a new internal standard to quantify chlorophylls in vegetable foods and to detect chlorophyll degradation products. The technique was applied to chlorophyll analysis of a selection of vegetable foods. Pigments were extracted with N,N-dimethylformamide from the vegetables and purified by solid phase extraction. Chlorophyll a, a', b, b', corresponding pheophytins, and zinc-phthalocyanine were separated by HPLC using a C-18 reverse-phase column and fluorescence detection. (C) 2003 Elsevier B.V. All rights reserved.
Abstract: We have evaluated urinary monitoring and erythrocyte analysis to determine Mg absorption in human subjects as alternatives to the conventional technique of faecal monitoring by stable-isotope techniques. Ten healthy adults received 2.2 mmol Mg-25 in water, together with wheat bread, followed 15 min later by intravenous injection of 0.6 mmol Mg-26 (day 1). Brilliant blue and Yb (given on day 0 and day 1 respectively) served as qualitative and quantitative faecal markers. Urine was collected for 6 d after test meal intake. Complete collections of faeces were made until excretion of the second brilliant blue marker (given on day 7). Mg isotope ratios were determined by thermal ionisation-MS in urine and faeces and by inductively coupled plasma-MS in erythrocytes. Absorption was determined based on: (1) 6 d urine pools; (2) 24 h urine pools (collected 22-46 h after test meal intake); (3) erythrocytes from a blood sample drawn on day 14; (4) complete 6 d faecal pools; (5) faecal pools based on the first three consecutive stools after excretion of the first brilliant blue marker. Differences in mean Mg absorption (42 44 %) were statistically insignificant between techniques, except when based on 6 d urine pools for which the value was significantly lower (33 (SD 7) %, P=0.0003, ANOVA). The results indicate that Mg absorption can be determined from 24 h urine pools or erythrocytes obtained 14 d after test meal intake, an alternative method to the more time-consuming and labour-intense faecal monitoring. The choice of technique depends on practical and financial considerations.
