Impacts of monosaccharide composition on immunomodulation by cello-pentaose, manno-pentaose, and xylo-pentaose: Unraveling the underlying molecular mechanisms.

Different types of functional oligosaccharides exhibit varying degrees of immune-enhancing effects, which might be attributable to differences in their glycosyl structures. The differences in the immunomodulatory action of three functional oligosaccharides with distinct glycosyl compositions: cello-oligosaccharides (COS), manno-oligosaccharides (MOS), and xylo-oligosaccharides (XOS), were investigated in mouse-derived macrophage RAW264.7. Moreover, the immune enhancement mechanism of oligosaccharides with diverse glycosyl compositions was investigated from a molecular interaction perspective. The TLR4-dependent immunoregulatory effect of functional oligosaccharides was shown by measuring the levels of tumor necrosis factor (TNF)-α and interleukin (IL)-6 in RAW264.7 cells treated with different functional oligosaccharides, both with and without Resatorvid [TAK-242] (a Toll-like receptor 4 [TLR4] inhibitor). Western blot analysis showed that binding of the three oligosaccharides to TLR4 activated the downstream signaling pathway and consequently enhanced the immune response. The fluorescence spectra and molecular docking results revealed that the main mechanisms by which these oligosaccharides attach to the TLR4 active pocket are hydrogen bonds and van der Waals forces. Functional oligosaccharides were ranked according to their affinity for TLR4, as follows: MOS > COS > XOS, indicating that oligosaccharides or polysaccharides containing mannose units may confer significant advantages for immune enhancement.

The Increase of Incomplete Degradation Products of Galactomannan Production by Synergetic Hydrolysis of ß-Mannanase and α-Galactosidase.

An integrated process to increase the yield of incomplete degradation products of galactomannan (GalM) especially for galactomanno-oligosaccharides (GalMOS) was suggested. Trichoderma reesei employed Avicel or GalMOS as a carbon source to produce ß-mannanase or α-galactosidase independently, with a result of 3.78 ± 0.12 U/mL of ß-mannanase activity and 2.45 ± 0.06 U/mL of α-galactosidase activity which were obtained, respectively. GalM in Sesbania seed was hydrolyzed simultaneously by a mixture of crude enzyme with ß-mannanase and α-galactosidase at a dosage of 20 U/g GalM and 15 U/g GalM, respectively; the yields of incomplete degradation products of GalM (IDP-GalM) and GalMOS were 78.84% ± 3.14% and 30.94% ± 0.38%, respectively, which was beneficial to improve the biological activity of the incomplete degradation products. The role of α-galactosidase addition in mixture enzymes is to remove the galactose substituents from mannan backbone of GalM and alleviate the steric hindrance of ß-mannanase hydrolysis.

Synthesis, antiproliferative activities, and DNA binding of coumarin-3-formamido derivatives.

Ten coumarin-3-formamido derivatives, N-benzyl-coumarin-3-carboxamide (2), N-fluorobenzyl-coumarin-3-carboxamide (3-5), N-methoxybenzyl-coumarin-3-carboxamide (6-8), N-((1-methyl-1H-imidazol-5-yl)methyl)-coumarin-3-carboxamide (9), N-(thiophen-2-ylmethyl)-coumarin-3-carboxamide (10), and N-(furan-2-ylmethyl)-coumarin-3-carboxamide (11), were synthesized and characterized. Compound 5 crystallizes in a monoclinic system P21 /c space group with four chemical formulas in a unit cell; molecules of compound 5 are self-assembled into a two-dimensional supramolecular structure by intermolecular hydrogen bonds and C⋯C π stacking. The potential anticancer effects of these compounds on HeLa (cervical carcinoma), MCF-7 (breast), A549 (lung), HepG2 (liver), and human umbilical vein (HUVEC) cells were examined. Compared with compounds 1-8 and 10-11, compound 9 exhibits potent in vitro cytotoxicity against HeLa cells and lower cytotoxicity against normal cells. Therefore, further in-depth investigations of compound 9 were performed. Absorption titration experiments and fluorescence spectroscopy studies suggested that compound 9 binds to DNA through the intercalation mode.

Synthesis and high antiproliferative activity of dehydroabietylamine pyridine derivatives in vitro and in vivo.

Several bioactive dehydroabietylamine Schiff-bases (L1-L4), amides (L5-L11) and complex CuL3(NO3)2, Cu(L5)3, Co(L6)2Cl2 had been synthesized successfully for developing more efficient but lower toxic antiproliferative compounds. Their antiproliferative activities to Hela (cervix), HepG2 (liver), MCF-7 (breast), A549 (lung) and HUVEC (umbilical vein, normal cell) were investigated in vitro. The toxicity of all compounds was less than dehydroabietylamine (L0). For HepG2 cells, L1, L2 and L3 had higher anti-HepG2 activity, especially L1 (0.52 µM) had highest anti-HepG2 activity but low toxicity. For MCF-7 cells, L1, L2, L3 and L4 had higher anti-MCF-7 activity, especially L3(0.49 µM) had highest anti-MCF-7 activity but low toxicity. For A549 cells, L2 and L3 had higher anti-A549 activity. Furthermore, L1 and L3 may be the great promise antiproliferative drugs with nontoxic side effects, due to the high anti-HepG2 and anti-MCF-7 inhibition rate in vivo, 65% and 61%, respectively. L1, L2 and L3 could induce apoptosis through intercalating into DNA.

Synthesis of novel, DNA binding heterocyclic dehydroabietylamine derivatives as potential antiproliferative and apoptosis-inducing agents.

Several dehydroabietylamine derivatives containing heterocyclic moieties such as thiophene and pyrazine ring were successfully synthesized. The antiproliferative activities of these thiophene-based Schiff-bases, thiophene amides, and pyrazine amides were investigated in vitro against Hela (cervix), MCF-7 (breast), A549 (lung), HepG2 (liver), and HUVEC (umbilical vein) cells by MTT assay. The toxicity of L1-L10 (IC50 = 5.92- >100 µM) was lower than L0 (1.27 µM) and DOX (4.40 µM) in every case. Compound L1 had higher anti-HepG2 (0.66 µM), anti-MCF-7 (5.33 µM), and anti-A549 (2.11 µM) and compound L3 had higher anti-HepG2 (1.63 µM) and anti-MCF-7 (2.65 µM) activities. Both of these compounds were recognized with high efficiency in apoptosis induction in HepG2 cells and intercalated binding modes with DNA. Moreover, with average IC50 values of 0.66 and 5.98 µM, L1 was nine times more effective at suppressing cultured HepG2 cells viability than normal cells (SI = 9). The relative tumor proliferation rate (T/C) was 38.6%, the tumor inhibition rate was up to 61.2%, which indicated that L1 had no significant toxicity but high anti-HepG2 activity in vivo. Thus, it may be a potential antiproliferation drug with nontoxic side effects.

Protein content and amino acids profile in 10 cultivars of ginkgo (Ginkgo biloba L.) nut from China.

As a traditional food and medicine source, ginkgo (Ginkgo biloba L.) nut is popularly consumed in East Asia. The aim of this work is to characterize protein content and amino acids profile in 10 ginkgo nut cultivars, named successively as no. 1 to no. 10. There were observed differences among the cultivars with respect to the contents of protein and amino acids, except Cys. The no. 6 cultivar presented the highest protein content (22.1 g/100 g DW), while the no. 9 had the lowest (16.2 g/100 g DW). The contents of EAA and NEAA were revealed to vary in the range of 14.3-26.2 and 21.4-41.1 g/100 g protein, respectively. The most abundant EAA was Leu, and the first limiting amino acid was Lys. The level of Arg was attractive, especially in the no. 5 cultivar (1741 mg/100 g DW) where it is comparable to hazelnut and pistachio. As confirmed by AAS and EAAI, the no. 5 cultivar presented the best amino acids profile and protein quality among these cultivars. These results have relevance to the scientific development and application of ginkgo nuts in the food industry.

Isolation, characterization and in vitro anticancer activity of an aqueous galactomannan from the seed of Sesbania cannabina.

As a pioneer plant, Sesbania cannabina is cultivated on a large scale for saline-alkali land improvement in China. Here, a native galactomannan (GalM) and a series of its hydrolysates (GalM40, GalM50 and GalM65) were obtained from S. cannabina seed by water extraction, ß-mannanase hydrolysate and ethanol precipitation. As elucidated by HPAEC, NMR, FT-IR and HPGPC, GalM was characterized as a ß-1,4-linked d-mannose polymer with single α-1,6-linked d-galactose side chain in a 2.4:1M ratio, and had a molecular weight of 1.42×106Da. MTT assay showed that these four fractions had significant inhibitory effect on A549, Hela, HepG2 and MCF-7 in a dose-dependent manner, and that GalM40 with second-highest MW (1.47×104Da) possessed higher activity amongst those fractions. Such strong inhibition effect on the growth of human cancer cells might derive from its ability to increase caspase-12 expression, as revealed from immunohistochemical analysis. In sum, this paper characterizes the molecular structure of S. cannabina galactomannan with anticancer activity, and this new knowledge will provide a basis for its further structure-properties research and ultimately, develop new possibilities for its use in high-value applications, such as functional food.

High anticancer potency on tumor cells of dehydroabietylamine Schiff-base derivatives and a copper(II) complex.

Five bioactive dehydroabietylamine Schiff-base derivatives (L1-L5) had been synthesized from Dehydroabietylamine (L0), and the complex Cu(L1)2 had been obtained from the compound L1 and copper(II) acetate. Their activities against Hela (cervix), MCF-7 (breast), A549 (lung), HepG2 (liver) and HUVEC (umbilical vein, normal cell) in vitro were investigated. The toxicity of L1-L5 and Cu(L1)2 was all lower than L0. For MCF-7 cell, L1, L3, L4, L5 and Cu(L1)2 had higher antitumor activity than L0. The smallest IC50 value was 2.58 µM of L5. For A549 cell, the IC50 value of the compound L4 was smaller than L0, which indicated that the compound L4 had higher anti-A549 activity than L0. For HepG2 cell, the IC50 value of L4(0.24 µM) and L5 (0.14 µM) were much smaller than L0, which suggested L4 and L5 had higher anti-HepG2 activity. L5 was 180 times more effective at inhibiting cultured HepG2 cells survival than normal cells, with average IC50 values of 0.14 and 25.56 µM. Furthermore, L0, L4 and L5 contrasting with Doxorubicin had been measured with the ability to induce apoptosis. It turned out that L4 and L5 could induce more HepG2 cells apoptosis, which suggested they may be potential antitumor drugs.

Synthesis and potential antineoplastic activity of dehydroabietylamine imidazole derivatives.

To seek more efficient and lower toxicity anticancer compounds, several imidazole combining dehydroabietylamine derivatives including organic salts (L 1 -L 2 ) and amides (L 3 -L 5 ) were synthesized. Their antineoplastic activity against HeLa (cervix), MCF-7 (breast), A549 (lung) and HepG2 (liver) cells and HUVECs (umbilical vein, normal cells) in vitro were evaluated by MTT assay. The results unequivocally showed that nearly all compounds had better antineoplastic activity and lower toxicity than dehydroabietylamine (L 0 ). For MCF-7 cells, L 2 (0.75 µM) and L 5 (2.17 µM) had higher anti-MCF-7 activity than L 0 and DOX. For A549 cells, L 1 (1.85 µM) and L 2 (4.37 µM) had higher anti-A549 activity than L 0 ; in particular, the IC50 value of L 1 was much lower than that of DOX. Among these investigated compounds, L 2 and L 5 had lower IC50 values (0.75 µM and 2.17 µM) against MCF-7 cells and lower toxicity, which suggested that they may be potential future anticancer drugs. In addition, L 1 and L 2 could suppress cancer cell proliferation by inducing apoptosis. L 1 -L 5 could bind with DNA through intercalation.

Characterization of non-methane hydrocarbons and their sources in an industrialized coastal city, Yangtze River Delta, China.

Ningbo is a highly industrialized city in the coastal area of the Yangtze River Delta (YRD), China. Large emissions and transport of non-methane hydrocarbons (NMHCs) may contribute to regional ozone (O3) and particulate matter (PM) pollution; however, the concentrations and sources of ambient NMHCs have not yet been investigated in Ningbo. In this study, ambient NMHCs were measured at two residential (SZ and CX) and two industrial (ZH and BL) sites and one suburban (XS) site over ten consecutive days in each season (10-20 December 2012 in winter, 14-23 April 2013 in spring, 15-24 July 2013 in summer, 22-31 October 2013, in autumn). A positive matrix factorization (PMF) model using multiple site data was deployed to explore the source contributions and their spatial and seasonal characteristics. The measurement results showed obvious seasonal variations in ambient NMHC concentrations (ranging from 17.89-28.48ppbv); chemical compositions were similar among the five sampling sites. PMF analysis showed that the petrochemical industry was the largest contributor (an average of 35.64%) to ambient NMHCs, while contributions of smaller sources (i.e., chemical and paint industries [14.34%], fuel and tank evaporation [16.02%], and residential solvent usage [7.24%]) showed spatial variations. Liquefied petroleum gas and fuel and tank evaporation contributed more in summer and autumn, while the contribution of the chemical and paint industries was greater in spring and winter. An evaluation of the ozone formation potential and secondary organic aerosol potential suggested that petrochemical and solvent-related sources were key parameters in mitigation of secondary pollutant formation. Seasonal variations in source contributions should be considered when formulating an effective NMHC abatement strategy.

Salecan Enhances the Activities of ß-1,3-Glucanase and Decreases the Biomass of Soil-Borne Fungi.

Salecan, a linear extracellular polysaccharide consisting of ß-1,3-D-glucan, has potential applications in the food, pharmaceutical and cosmetic industries. The objective of this study was to evaluate the effects of salecan on soil microbial communities in a vegetable patch. Compositional shifts in the genetic structure of indigenous soil bacterial and fungal communities were monitored using culture-dependent dilution plating, culture-independent PCR-denaturing gradient gel electrophoresis (DGGE) and quantitative PCR. After 60 days, soil microorganism counts showed no significant variation in bacterial density and a marked decrease in the numbers of fungi. The DGGE profiles revealed that salecan changed the composition of the microbial community in soil by increasing the amount of Bacillus strains and decreasing the amount of Fusarium strains. Quantitative PCR confirmed that the populations of the soil-borne fungi Fusarium oxysporum and Trichoderma spp. were decreased approximately 6- and 2-fold, respectively, in soil containing salecan. This decrease in the amount of fungi can be explained by salecan inducing an increase in the activities of ß-1,3-glucanase in the soil. These results suggest the promising application of salecan for biological control of pathogens of soil-borne fungi.

Process-specific emission characteristics of volatile organic compounds (VOCs) from petrochemical facilities in the Yangtze River Delta, China.

Process-specific emission characteristics of volatile organic compounds (VOCs) from petrochemical facilities were investigated in the Yangtze River Delta, China. Source samples were collected from various process units in the petrochemical, basic chemical, and chlorinated chemical plants, and were measured using gas chromatography-mass spectrometry/flame ionization detection. The results showed that propane (19.9%), propene (11.7%), ethane (9.5%) and i-butane (9.2%) were the most abundant species in the petrochemical plant, with propene at much higher levels than in petrochemical profiles measured in other regions. Styrene (15.3%), toluene (10.3%) and 1,3-butadiene (7.5%) were the major species in the basic chemical industry, while halocarbons, especially dichloromethane (15.2%) and chloromethane (7.5%), were substantial in the chlorinated chemical plant. Composite profiles were calculated using a weight-average approach based on the VOC emission strength of various process units. Emission profiles for an entire petrochemical-related industry were found to be process-oriented and should be established considering the differences in VOC emissions from various manufacturing facilities. The VOC source reactivity and carcinogenic risk potential of each process unit were also calculated in this study, suggesting that process operations mainly producing alkenes should be targeted for possible controls with respect to reducing the ozone formation potential, while process units emitting 1,3-butadiene should be under priority control in terms of toxicity. This provides a basis for further measurements of process-specific VOC emissions from the entire petrochemical industry. Meanwhile, more representative samples should be collected to reduce the large uncertainties.

Daily variation in global and local DNA methylation in mouse livers.

DNA methylation is one of the best-characterized epigenetic modifications and has an important biological relevance. Here we showed that global DNA methylation level in mouse livers displayed a daily variation where the peak phases occurred during the end of the day and the lowest level at the beginning of the day in the light-dark or dark-dark cycles. Typical repeat sequence long interspersed nucleotide element-1 (LINE-1) had a similar methylation rhythm to global DNA. DNA methyltransferase 3A (DNMT3A) and ratio of S-adenosylmethionine (SAM) to S-adenosylhomocysteine (SAH) brought a relative forward daily variation to global DNA methylation, and the temporary change in ratio of SAM to SAH had no influence on the DNA methylation level. The rhythm of global DNA methylation was lost and DNA methylation level was increased in Per1-/-Per2-/- double knockout mice, which were in accordance with changes of Dnmt3a mRNA levels and its rhythm. Our results suggest that the daily variation in global DNA methylation was associated with the change of Dnmt3a expression rather than ratio of SAM to SAH.

Salecan diet increases short chain fatty acids and enriches beneficial microbiota in the mouse cecum.

Salecan, a linear extracellular polysaccharide consisting of ß-(1,3)-D-glucan, has potential applications in the food industry due to its excellent toxicological profile and rheological properties. The aim of the present study was to evaluate the effects of dietary supplementation with 8% Salecan on the gastrointestinal microbiota in mice. In the Salecan group, the following significant differences (p<0.05) from the cellulose group were found: increased body weight gain, greater mass of cecum and cecal contents, and higher butyrate concentrations in the cecal and colonic contents at wk 4. Moreover, populations of Lactobacillus and Bifidobacterium increased 3- and 6-fold, respectively, in the cecal contents of mice consuming Salecan. These results suggest that the dietary incorporation of Salecan, by providing SCFAs and increasing beneficial microbiota, may be beneficial in improving gastrointestinal health, and have relevance to the use of Salecan as a dietary supplement for human consumption.

Supplementation of the diet with Salecan attenuates the symptoms of colitis induced by dextran sulphate sodium in mice.

As a water-soluble extracellular ß-glucan produced by Agrobacterium sp. ZX09, Salecan has an excellent toxicological profile and exerts multiple physiological effects. The aims of the present study were to investigate the protective effects of a Salecan diet in the well-defined dextran sulphate sodium (DSS) model of experimental murine colitis and to elucidate the mechanism involved in its effects with special attention being paid to its effect on the production of TNF-α, a primary mediator involved in the inflammatory response. Male C57BL/6J mice were fed a diet supplemented with either 4 or 8 % Salecan for 26 d and DSS was administered to induce acute colitis during the last 5 d of the experimental period. Several clinical and inflammatory parameters as well as mRNA expression of TNF-α and Dectin-1 were evaluated. The results indicated that the dietary incorporation of Salecan attenuated the severity of DSS colitis as evidenced by the decreased disease activity index, reduced severity of anaemia, attenuated changes in colon architecture and reduced colonic myeloperoxidase activity. This protection was associated with the down-regulation of TNF-α mRNA levels, which might derive from its ability to increase Dectin-1 mRNA levels. In conclusion, the present study suggests that Salecan contributes to the reduction of colonic damage and inflammation in mice with DSS-induced colitis and holds promise as a new, effective nutritional supplement in the management of inflammatory bowel disease.

Endogenous A1 adenosine receptor protects mice from acute ethanol-induced hepatotoxicity.

Previous studies have indicated a critical role of adenosine and its receptors in the pathogenesis of liver diseases. The aim of this study was to determine the contribution of A1 adenosine receptor (A1AR) to acute ethanol-induced hepatotoxicity. Wild-type (WT) and A1AR(-/-) mice were intragastrically administered with ethanol (5 g/kg), and hepatic injury was evaluated 6h thereafter. Mice lacking A1AR were more susceptible to ethanol-induced liver damage than WT mice, as evidenced by higher serum transaminase levels and increased extent of histopathological changes. Ethanol induced triglycerides accumulation in the serum and liver, and this accumulation was augmented in A1AR(-/-) mice. Analysis of gene expression in the liver revealed up-regulated mRNA levels of genes related to lipogenesis (including: FAS, SCD1, ACC1, DGAT2, and PPARγ) in A1AR(-/-) mice after ethanol treatment. In addition, lack of A1AR aggravated lipid peroxidation and superoxide dismutase depletion caused by acute ethanol exposure. A subsequent study revealed that, pretreatment with A1AR antagonist DPCPX increases the sensitivity of mice to ethanol-induced liver injury. In conclusion, these results indicated that endogenous A1AR activation protects mice against acute ethanol-induced liver injury by reducing oxidative stress and decreasing lipid accumulation.

Laxative effects of Salecan on normal and two models of experimental constipated mice.

BACKGROUND: Constipation is one of the most common gastrointestinal complaints with a highly prevalent and often chronic functional gastrointestinal disorder affecting health-related quality of life. The aim of the present study was to evaluate the effects of Salecan on fecal output and small intestinal transit in normal and two models of drug-induced constipation mice. METHODS: ICR mice were administrated intragastrically (i.g.) by gavage with 100, 200 and 300 mg/kg body weight (BW) of Salecan while the control mice were received saline. The constipated mice were induced by two types of drugs, loperamide (5 mg/kg BW, i.g.) and clonidine (200 µg/kg BW, i.g.), after Salecan treatment while the control mice were received saline. Number, weight and water content of feces were subsequently measured. Small intestinal transit was monitored by phenol red marker meal. RESULTS: Salecan (300 mg/kg BW) significantly increased the number and weight of feces in normal mice. In two models of drug-induced constipation, Salecan dose-dependently restored the fecal number and fecal weight. The water content of feces was markedly affected by loperamide, but not by clonidine. Treatment with Salecan significantly raised the fecal water content in loperamide-induced constipation mice. Moreover, Salecan markedly stimulated the small intestinal transit in both loperamide- and clonidine-induced constipation model mice. CONCLUSIONS: These results suggest that Salecan has a potential to be used as a hydrophilic laxative for constipation.

Loss of A(1) adenosine receptor attenuates alpha-naphthylisothiocyanate-induced cholestatic liver injury in mice.

Cholestasis has limited therapeutic options and is associated with high morbidity and mortality. The A(1) adenosine receptor (A(1)AR) was postulated to participate in the pathogenesis of hepatic fibrosis induced by experimental extrahepatic cholestasis; however, the contribution of A(1)AR to intrahepatic cholestatic liver injury remains unknown. Here, we found that mice lacking A(1)AR were resistant to alpha-naphthyl isothiocyanate (ANIT)-induced liver injury, as evidenced by lower serum liver enzyme levels and reduced extent of histological necrosis. Bile acid accumulation in liver and serum was markedly diminished in A(1)AR(-/-) mice compared with wild-type (WT) mice. However, biliary and urinary outputs of bile acids were significantly enhanced in A(1)AR(-/-) mice. In the liver, mRNA expression of genes related to bile acid transport (Bsep and Mdr2) and hydroxylation (Cyp3a11) was increased in A(1)AR(-/-) mice. In the kidney, A(1)AR deficiency prevented the decrease of glomerular filtration rate caused by ANIT. Treatment of WT mice with A(1)AR antagonist DPCPX also protected against ANIT hepatotoxicity. Our results indicated that lack of A(1)AR gene protects mice from ANIT-induced cholestasis by enhancing toxic biliary constituents efflux through biliary excretory route and renal elimination system and suggested a potential role of A(1)AR as therapeutic target for the treatment of intrahepatic cholestasis.

Loss of mPer2 increases plasma insulin levels by enhanced glucose-stimulated insulin secretion and impaired insulin clearance in mice.

The existence of peripheral oscillators has been shown, and they are critically important for organizing the metabolism of the whole body. Here we show that mice deficient in mPer2 markedly increase circulatory levels of insulin compared with wild type mice. Insulin secretion was more effectively stimulated by glucose, and alloxan, a glucose analogue, induced more severe hyperglycemia in mPer2-deficient mice. Hepatic insulin degrading enzyme (Ide) displayed an obvious day and night rhythm, which was impaired in mPer2-deficient mice, leading to a decrease in insulin clearance. Deficiency in mPer2 caused increased Clock expression and decreased expression of Mkp1 and Ide1, possibly underlying the observed phenotypes and suggesting that mPer2 plays a role in regulation of circulating insulin levels.

Results of a 90-day safety assessment study in mice fed a glucan produced by Agrobacterium sp. ZX09.

Salecan is a novel water-soluble glucan produced by Agrobacterium sp. ZX09. It has potential application as a food additive with a unique chemical composition and excellent physicochemical properties. The objective of this study was to investigate the acute and subchronic toxicity of Salecan. The oral LD50 of Salecan in ICR mice was greater than 3000 mg/kg body weight. In the subchronic study, ICR mice (10/sex/group) were fed diets containing 0%, 1.0%, 2.5% and 5.0% of Salecan (weight/weight) for 13 weeks. Based on the results from the subchronic study, the overall health, body weight gain, food consumption and clinical pathology parameters were comparable between the groups feed Salecan and the control. No dose-related effects were observed in the treated animals. The only exception was the observation that blood glucose in female mice fed Salecan was lower than in the control group. In addition, the fecal matter from Salecan fed mice exhibited increased water content versus the control animals. The no observed adverse effect level (NOAEL) of 14478 mg/kg body weight/day was determined. The results from this study support the conclusion that Salecan is non-toxic at the levels tested and does not pose a risk to human health when used in food.