Siriporn Tuntipopipat

Iron absorption by human subjects from different iron fortification compounds added to Thai fish sauce

Objectives:(a) To measure iron absorption by human subjects from citric acid stabilized fish sauce fortified with ferrous sulfate, ferric ammonium citrate or ferrous lactate and (b) to identify the effect of added citric acid (3 g/l) on iron absorption from ferrous sulfate fortified fish sauce.Design:Iron absorption from the intrinsically labeled compounds was determined via erythrocyte incorporation of isotopic labels (57Fe and 58Fe) using a randomized crossover design. In three separate absorption studies, 10 adult women each consumed a basic test meal of rice and vegetable soup seasoned with isotopically labeled, iron fortified fish sauce.Results:Iron absorption was significantly lower from ferrous lactate and from ferric ammonium citrate fortified fish sauce than from ferrous sulfate fortified fish sauce. Fractional iron absorption (geometric mean; −1s.d., +1s.d.) was 8.7(3.6; 21.4)% for ferrous lactate compared to 13.0(5.4; 31.4)% from ferrous sulfate, P=0.003 (study 1) and 6.0(2.5; 14.3)% from ferric ammonium citrate relative to 11.7(4.4; 30.7)% from ferrous sulfate, P<0.001, in study 2. Citric acid added at a molar ratio of ∼2.5 to iron had no effect on iron absorption from ferrous sulfate (study 3). Iron absorption in the presence of citric acid was 14.1(6.4; 30.8)% compared to 12.0(5.8; 24.7)% in its absence (P=0.26).Conclusions:Iron absorption was 50–100% higher from ferrous sulphate fortified fish sauce than from fish sauce fortified with ferric ammonium citrate or ferrous lactate. In the presence of citric acid as a chelator, ferrous sulfate would appear to be a useful fortificant for fish sauce.Sponsorship:International Atomic Energy Agency (IAEA), Vienna, Austria

Anti-Inflammatory Activities of Extracts of Thai Spices and Herbs with Lipopolysaccharide-Activated RAW 264.7 Murine Macrophages

Nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha) play important roles in inflammatory processes. This study examined whether 13 spices/herbs commonly used in Thai dishes modulate the production of NO and TNF-alpha by the RAW 264.7 mouse macrophage cell line pretreated with plant extracts (1-100 microg/mL) prior to activation by bacterial lipopolysaccharide (LPS). Tested plant tissues were extracted with ethanol with the exception of roselle, which was extracted with 70% acetone. Eight of the 13 plant extracts inhibited NO and TNF-alpha production in a dose-dependent manner without exerting cytotoxicity. Extract from Limnophila aromatica (Kyeng) was the most robust suppressor of NO production, followed by dill, kaffer lime, chili, Teaw, mint, sweet basil, and pea eggplant, respectively (range of 50% inhibitory concentration [IC(50)] = 11.4-74.6 microg/mL). Kyeng also exhibited the greatest inhibition of TNF-alpha production (IC(50) = 10.5 microg/mL). IC(50) values for NO and TNF-alpha production in LPS-activated RAW 264.7 cells for these extracts were highly correlated (r = 0.772, P = .025). These results suggest that extracts from some spices/herbs in the habitual Thai diet possess anti-inflammatory activity. Moreover, the results support the use of NO production in LPS-activated RAW 264.7 cells as a rapid and cost-effective tool for screening the anti-inflammatory activity of extracts of spices/herbs.

Inhibitory effects of spices and herbs on iron availability

Spices and herbs are extensively used in indigenous diets in tropical regions where prevalence of iron deficiency is still high. They are rich in polyphenolic compounds that are expected to inhibit iron absorption by forming iron complexes in the intestine, making dietary iron less available for absorption. The effects of six spices and herbs (chili pepper, garlic, ‘Pak kyheng’ (Thai leafy vegetable), shallot, tamarind, turmeric) and one mixture of spices (curry paste) on iron availability were determined by measuring the percentage dialyzable iron after addition of spices and herbs to a rice meal after simulated digestion. All tested spices and herbs contained from 0.5 to 33 mg polyphenol per meal and were potent inhibitors of iron availability (20–90%), reducing iron availability in a dose-dependent manner—with the exception of tamarind, which at 11 mg polyphenol per meal enhanced iron availability. Our findings demonstrate that culinary spices and herbs can play an important role in iron nutrition.

Eryngium foetidum suppresses inflammatory mediators produced by macrophages.

OBJECTIVE This study assessed anti-inflammatory and antioxidant activities of E. foetidum leaf extract on LPS-activated murine macrophages. METHODS RAW264.7 cells were pretreated with or without E. foetidum extract for 1 h prior to incubation with LPS for 24 h. Anti-inflammatory activity was evaluated with reference to iNOS, COX-2, TNF-α and IL-6 gene expression. In addition, NO and intracellular ROS generation were determined by Griess method and fluorescence intensity and activation of MAPKs and IκB by Western blotting. RESULTS Prior treatment with E. foetidum leaf extract inhibited elevation of IL-6, TNF-α, iNOS and COX-2, together with their cognate mRNAs in a dose-dependent manner. NO and intracellular ROS contents were similarly reduced. These effects were due to inhibition of LPS-induced phosphorylation of JNK and p38 as well as IκB. E. foetidum ethanol extract was shown to contain lutein, β-carotenes, chlorogenic acid, kaempferol and caffeic acid, compounds known to exert these bioactive properties. CONCLUSIONS E. foetidum leaf extract possesses suppressive effects against pro-inflammatory mediators. Thus, E. foetidum has a high potential to be used as a food supplement to reduce risk of cancer associated with inflammation.

Calcium Absorption from Commonly Consumed Vegetables in Healthy Thai Women

The absorbability of calcium from ivy gourd, a green leafy vegetable (Coccinia grandix Voigt.) and winged bean young pods (Psophocarpus tetragonolobus [L] DC) were measured in 19 healthy adult women aged 20 to 45 y, in a 3-way, randomized-order, crossover design with an average calcium load of 100 mg and milk as the referent. The test meals were extrinsically labeled with 44Ca and given with rice as breakfast after an overnight fast. Absorption of calcium was determined on a blood sample drawn 5 h after ingestion of the test meal. Fractional calcium absorption (X+/- SD) was 0.391 +/- 0.128 from winged beans, 0.476 +/- 0.109 from ivy gourd, and 0.552 +/- 0.119 from milk. The difference in fractional calcium absorption for these 2 vegetables was significant (P < 0.05) and the fractional calcium absorption from these 2 vegetables were both significantly lower than from milk. The difference was partly accounted for by the phytate, oxalate, and dietary fiber content of the vegetables. However, calcium bioavailability of these 2 vegetables, commonly consumed among Thais, was relatively good compared to milk (71% to 86% of milk) and could be generally recommended to the public as calcium sources other than milk and Brassica vegetables.

Anti-inflammatory activities of red curry paste extract on lipopolysaccharide-activated murine macrophage cell line

OBJECTIVE This study investigated anti-inflammatory and antioxidant activities of an ethanol extract from Thai red curry paste. METHODS The RAW264.7 murine macrophage cell line was incubated with the extract (65-260 μg/mL) with or without lipopolysaccharide. The anti-inflammatory activities of the extract were examined by measuring inducible nitric oxide synthase, cyclo-oxygenase-2, tumor necrosis factor-α, and interleukin-6 mRNA and protein level by reverse transcription-polymerase chain reaction, western blotting, and enzyme-linked immunosorbent assay, respectively. Nitric oxide production and intracellular reactive oxygen species generation were determined by the Griess method and fluorescence intensity. The activation of mitogen-activated protein kinases and inhibitor κB were determined by western blot. RESULTS Exposure of cells with the extract significantly suppressed lipopolysaccharide-induced nitric oxide production and inducible nitric oxide synthase, cyclo-oxygenase-2, tumor necrosis factor-α, and interleukin-6 expressions (P < 0.05) by dose-dependently without cytotoxic effect. Intracellular reactive oxygen species significantly decreased (P < 0.05) in lipopolysaccharide-induced RAW264.7 cells. The inhibitory effect was mediated partly by inhibiting activation of inhibitor κB-α and mitogen-activated protein kinases. CONCLUSION These results suggest that the anti-inflammatory and antioxidant properties of Thai red curry paste stem from bioactive compounds present in the spice and herb constituents. The health benefits of Thai red curry paste warrant further investigations in vivo.

A simplified method for the fractionation of Gnathostoma -specific antigens for serodiagnosis of human gnathostomosis

Specific immunoreactive components present in crude somatic extract and in excretory-secretory (ES) products of Gnathostoma spinigerum advanced third-stage larvae (L3) were identified by Western blotting and their diagnostic potential evaluated by indirect ELISA. Although both crude antigen preparations were highly complex, the ES antigen gave a more satisfactory diagnostic result. Most G. spinigerum specific components present in the somatic and ES preparations had molecular weights below 29 kD and were not glycosylated, judging from the concanavalin A staining pattern. Specific diagnostic antigens were prepared by subjecting the crude preparations to SDS-PAGE. Low molecular weight components were identified and electroeluted from the gel. Excess SDS was removed by the use of an ion retardation resin. The antigens obtained by this relatively simple procedure were found to be highly specific for G. spinigerum. Sensitivity, specificity, and positive and negative predictive values for the assay using the fractionated somatic antigen were 100%.

Anti-Inflammatory Activity of Bioaccessible Fraction from Eryngium foetidum Leaves

Eryngium foetidum (EF) has long been used as a medicinal plant and culinary spice in tropical regions. Phytochemicals in its leaves have been proposed to be responsible for the anti-inflammatory and antioxidant activities. The present study used in vitro digestion coupled with Caco-2 cells to assess such activities. Caco-2 cells were incubated with aqueous fraction from simulated digestion (bioaccessible fraction) of EF leaves with/without bile extract prior to stimulation with interleukin-1 beta (IL-1β). Monocyte chemoattractant protein-1 (MCP-1) and IL-8 in culture media and the intracellular reactive oxygen species (ROS) were measured. Approximately 24% β-carotene and 35% lutein of leaves were present in the aqueous fraction. The transfer of caffeic and chlorogenic acids to the aqueous fraction was 76%–81%, while that of kaempferol was 48%. Prior incubation of Caco-2 cells with the bioaccessible fraction suppressed IL-1β activated IL-8 and MCP-1 by 33%, but the fraction lacking mixed micelles decreased IL-8 and MCP-1 levels only by 11%. The pretreatment of Caco-2 cells with the bioaccessible fraction of EF reduced ROS by 34%; the fraction lacking mixed micelles decreased ROS by 28%. These data suggest that bioactive compounds partitioning in mixed micelles play a significant role to suppress the proinflammatory insult but with a modest antioxidant effect.

Anticlastogenic effect of Eryngium foetidum L. assessed by erythrocyte micronucleus assay.

The aim of this study was to investigate the anticlastogenicity as well as the clastogenicity of Eryngium foetidum leaf (EF) using the in vivo mouse peripheral blood erythrocyte micronucleus assay. Eighty ICR male mice were fed AIN-76 diet supplemented with ground freeze-dried EF at 0.0%, 0.8%, 1.6% and 3.2% for 2 weeks prior to the administration of both direct-acting, mitomycin C (MMC), and indirect-acting, 7, 12-dimethylbenz(a) anthracene (DMBA) clastogens. Peripheral blood samples were collected from mice just before administration of clastogen and at 24 and 48 h thereafter for MMC. Blood samples were collected at the same times and after 72 h for DMBA. Then, reticulocytes in blood samples were counted using fluorescent microscopy. The results indicated that EF had no clastogenic effect in mice. All doses of diets supplemented with EF decreased the number of micronucleated peripheral reticulocytes in all the MMC-treated groups in a dose dependent manner, but significant reduction was found only at 1.6% and 3.2% EF in the DMBA-treated groups. It can be concluded that EF has no clastogenicity, but possesses anticlastogenic potential against both direct- and indirect-acting types of clastogen in mice.

Ferulic Acid Supplementation Improves Lipid Profiles, Oxidative Stress, and Inflammatory Status in Hyperlipidemic Subjects: A Randomized, Double-Blind, Placebo-Controlled Clinical Trial

Ferulic acid is the most abundant phenolic compound found in vegetables and cereal grains. In vitro and animal studies have shown ferulic acid has anti-hyperlipidemic, anti-oxidative, and anti-inflammatory effects. The objective of this study is to investigate the effects of ferulic acid supplementation on lipid profiles, oxidative stress, and inflammatory status in hyperlipidemia. The study design is a randomized, double-blind, placebo-controlled trial. Subjects with hyperlipidemia were randomly divided into two groups. The treatment group (n = 24) was given ferulic acid (1000 mg daily) and the control group (n = 24) was provided with a placebo for six weeks. Lipid profiles, biomarkers of oxidative stress and inflammation were assessed before and after the intervention. Ferulic acid supplementation demonstrated a statistically significant decrease in total cholesterol (8.1%; p = 0.001), LDL-C (9.3%; p < 0.001), triglyceride (12.1%; p = 0.049), and increased HDL-C (4.3%; p = 0.045) compared with the placebo. Ferulic acid also significantly decreased the oxidative stress biomarker, MDA (24.5%; p < 0.001). Moreover, oxidized LDL-C was significantly decreased in the ferulic acid group (7.1%; p = 0.002) compared with the placebo group. In addition, ferulic acid supplementation demonstrated a statistically significant reduction in the inflammatory markers hs-CRP (32.66%; p < 0.001) and TNF-α (13.06%; p < 0.001). These data indicate ferulic acid supplementation can improve lipid profiles and oxidative stress, oxidized LDL-C, and inflammation in hyperlipidemic subjects. Therefore, ferulic acid has the potential to reduce cardiovascular disease risk factors.