The orphan transporter hippocampus-abundant transcript 1 (Hiat1) was first identified in the mammalian brain. Its specific substrate specificity, however, has not been investigated to date. Here, we identified and analyzed Hiat1 in a crustacean, the green crab Carcinus maenas. Our phylogenetic analysis showed that Hiat1 protein is conserved at a considerable level between mammals and this invertebrate (ca. 78% identical and conserved amino acids). Functional expression of Carcinus maenas Hiat1 in Xenopus laevis oocytes demonstrated the capability to transport ammonia (likely NH4+) in a sodium-dependent manner. Furthermore, applying quantitative polymerase chain reaction, our results indicated a physiological role for Carcinus maenas Hiat1 in ammonia homeostasis, as mRNA abundance increased in posterior gills in response to elevated circulating hemolymph ammonia upon exposure to high environmental ammonia. Its ubiquitous mRNA expression pattern also suggests an essential role in general cellular detoxification of ammonia. Overall, our results introduce a new ubiquitously expressed ammonia transporter, consequently demanding revision of our understanding of ammonia handling in key model systems from mammalian kidneys to crustacean and fish gills.

ABSTRACT Ammonia excretion in fish excretory epithelia is a complex interplay of multiple membrane transport proteins and mechanisms. Using the model system of zebrafish (Danio rerio) larvae, here we identified three paralogues of a novel ammonia transporter, hippocampus-abundant transcript 1 (DrHiat1), also found in most vertebrates. When functionally expressed in Xenopus laevis oocytes, DrHiat1a and DrHiat1b promoted methylamine uptake in a competitive manner with ammonia. In situ hybridization experiments showed that both transporters were expressed as early as the 4-cell stage in zebrafish embryos and could be identified in most tissues 4 days post-fertilization. Larvae experiencing morpholino-mediated knockdown of DrHiat1b exhibited significantly lower whole-body ammonia excretion rates compared with control larvae. Markedly decreased site-specific total ammonia excretion of up to 85% was observed in both the pharyngeal region (site of developing gills) and the yolk sac (region shown to have the highest NH4+ flux). This study is the first to identify DrHiat1b/DrHIAT1 in particular as an important contributor to ammonia excretion in larval zebrafish. Being evolutionarily conserved, these proteins are likely involved in multiple other general ammonia-handling mechanisms, making them worthy candidates for future studies on nitrogen regulation in fishes and across the animal kingdom.

Avenanthramides are phenolic compounds unique to oats and may contribute to health-promoting properties associated with oat consumption. This study used Xenopus laevis oocytes expressing the glucose transporters, glucose transporter 2 (GLUT2) or sodium-glucose transport protein 1 (SGLT1) and human Caco-2 cells models to investigate the effect of oat avenanthramides on human intestinal glucose transporters. The presence of avenanthramide reduced the glucose uptake in a dose-dependent manner in Caco-2 cells. Glucose uptake in oocytes expressing either GLUT2 or SGLT1 was nullified by oat avenanthramide. There was no significant difference between the inhibition potencies of avenanthramides C and B. Thus, our results suggest that avenanthramides may contribute to the antidiabetic properties of oats. Practical applications The present research focus on the antidiabetic properties of avenanthramides, which are unique phenolic compounds found in oats. Inhibiting the activities of the glucose transport proteins expressed in the small intestine is a known strategy to improve the control of postprandial glucose level. We therefore examined the inhibitory effects of avenanthramides on two glucose transporters, glucose transporter 2 and sodium-glucose transport protein 1, predominantly found in the small intestine using the human small intestinal cell model Caco-2 cell line and by heterologously expressing these two transporters in the Xenopus laevis oocytes. Based on our results, we have confirmed for the first time that the glucose uptake is indeed inhibited by the presence of avenanthramides, suggesting the possibility of incorporating avenanthramides in foods to enhance postprandial glucose response, and ultimately improve the management of diabetes. Therefore, future research could consider utilizing this evidence in the development of diabetic-friendly functional foods or nutraceuticals containing avenanthramides.

Data for figures in manuscript "Hiat1 – a new transporter involved in ammonia regulation"

Background: Reduced plasma vitamin C concentrations in chronic diseases may result from abnormal urinary excretion of vitamin C: a renal leak. We hypothesized that vitamin C renal leak may be associated with disease-mediated renal dysregulation, resulting in aberrant vitamin C renal reabsorption and increased urinary loss. Objectives: We investigated the prevalence, clinical characteristics, and genomic associations of vitamin C renal leak in Fabry disease, an X-linked lysosomal disease associated with renal tubular dysfunction and low plasma vitamin C concentrations.Methods: We conducted a non-randomized cross-sectional cohort study of men aged 24-42 y, with Fabry disease (n = 34) and controls without acute or chronic disease (n = 33). To match anticipated plasma vitamin C concentrations, controls were placed on a low-vitamin C diet 3 wk before inpatient admission. To determine the primary outcome of vitamin C renal leak prevalence, subjects were fasted overnight, and matched urine and fasting plasma vitamin C measurements were obtained the following morning. Vitamin C renal leak was defined as presence of urinary vitamin C at plasma concentrations below 38 & mu;M. Exploratory outcomes assessed the association between renal leak and clinical parameters, and genomic associations with renal leak using single nucleotide polymorphisms (SNPs) in the vitamin C transporter SLC23A1. Results: Compared with controls, the Fabry cohort had 16-fold higher odds of renal leak (6% vs. 52%; OR: 16; 95% CI: 3.30, 162; P < 0.001). Renal leak was associated with higher protein creatinine ratio (P < 0.01) and lower hemoglobin (P = 0.002), but not estimated glomerular filtration rate (P = 0.54). Renal leak, but not plasma vitamin C, was associated with a nonsynonymous single nucleotide polymorphism in vitamin C transporter SLC23A1 (OR: 15; 95% CI: 1.6, 777; P = 0.01).Conclusions: Increased prevalence of renal leak in adult men with Fabry disease may result from dysregulated vitamin C renal physiology and is associated with abnormal clinical outcomes and genomic variation.

Background: The consumption of 2 g/d plant sterols (PSs) reduces circulating LDL cholesterol by <= 10%. The degree of LDL cholesterol lowering was associated with specific apolipoprotein E [APOE, Reference SNP (rs)429358] and cholesterol 7 alpha-hydroxylase (CYP7A1. rs3808607) genosets in previous post hoc analyses of randomized controlled trials. However, because post hoc analyses do not conform to the randomization model, there is a greater potential that the findings could be due to type I error, thus warranting validation through an a priori-designed intervention trial. Objectives: The GenePredict Plant Sterol study (GPS) was designed to validate associations of LDL cholesterol lowering with specific APOE and CYP7A1 genosets through a priori recruitment of individuals carrying prespecified genosets. Methods: A 2-center, double-blind, placebo-controlled, randomized 2-period crossover dietary intervention with 2 g/d PS for 28 d with a minimum 28-d washout was undertaken from July 2017 to December 2019. A priori recruitment of individuals with slightly elevated LDL cholesterol was based on genosets of APOE isoforms and CYP7A1 rs3808607. Randomization was performed with stratification by sex and genoset. Results: The recruitment target of 64 participants with prespecified genosets could not be reached. despite the screening of 477 individuals; 42 participants completed the intervention trial. Reductions in LDL cholesterol were similar across all 3 genosets (-0.298 +/- 0.164, -0.357 +/- 0.115, -0.293 +/- 0.109 mmol/L: P = 0.0002 overall; P = 0.9126 for treatment x genoset), providing evidence that the shortfall in recruitment might not have stopped the trial from meeting the objective. Conclusions: APOE and CYP7A1 genotypes did not influence the efficacy of LDL, cholesterol reductions upon dietary intervention with PSs. Findings of previous post hoc analyses could not be validated in a trial using a priori genotype-based recruitment. Obtaining adequate numbers of participants is challenging in trials using genoset-based recruitment, even for common variants.

BACKGROUND:Blood lipid concentrations display high interindividual variability in response to dietary interventions, partly due to genetic factors. Existing studies have focused on single nucleotide polymorphisms (SNPs) analyzed individually, which only explain a limited fraction of the variability of these complex phenotypes.OBJECTIVE:We aimed to identify combinations of SNPs associated with the variability in LDL cholesterol and triglyceride (TG) concentration changes following 5 dietary interventions.DESIGN:In a multicenter randomized crossover trial, 92 participants with elevated waist circumference and low HDL cholesterol concentrations consumed 5 isoenergetic diets for 4 wk: a diet rich in saturated fatty acids (SFAs) from cheese, SFA from butter, monounsaturated fatty acids (MUFAs), n-6 polyunsaturated fatty acids (PUFAs), and a diet higher in carbohydrates (CHO). The association between 22 candidate SNPs in genes involved in lipid and bile acid metabolism and transport and changes in LDL cholesterol and TG concentrations was assessed with univariate statistics followed by partial least squares regression.RESULTS:Endpoint LDL cholesterol concentrations were significantly different (cheese: 3.18 ± 0.04, butter: 3.31 ± 0.04, MUFA: 3.00 ± 0.04, PUFA: 2.81 ± 0.04, CHO: 3.11 ± 0.04 mmol/L; P < 0.001) while endpoint TG concentrations were not (P = 0.117). Both displayed consistently elevated interindividual variability following the dietary interventions (CVs of 34.5 ± 2.2% and 55.8 ± 1.8%, respectively). Among the 22 candidate SNPs, only ABCA1-rs2066714 and apolipoprotein E (APOE) isoforms exhibited consistent significant effects, namely on LDL cholesterol concentrations. However, several SNPs were significantly associated with changes in LDL cholesterol and TG concentrations in a diet-specific fashion. Generated multivariate models explained from 16.0 to 33.6% of the interindividual variability in LDL cholesterol concentration changes and from 17.5 to 32.0% of that in TG concentration changes.CONCLUSIONS:We report combinations of SNPs associated with a significant part of the variability in LDL cholesterol and TG concentrations following dietary interventions differing in their fatty acid profiles.

Abstract Diets varying in SFA and MUFA content can impact glycaemic control; however, whether underlying differences in genetic make-up can influence blood glucose responses to these dietary fatty acids is unknown. We examined the impact of dietary oils varying in SFA/MUFA content on changes in blood glucose levels (primary outcome) and whether these changes were modified by variants in the stearoyl-CoA desaturase (SCD) gene (secondary outcome). Obese men and women participating in the randomised, crossover, isoenergetic, controlled-feeding Canola Oil Multicenter Intervention Trial II consumed three dietary oils for 6 weeks, with washout periods of ˜6 weeks between each treatment. Diets studied included a high SFA/low MUFA Control oil (36·6 % SFA/28·2 % MUFA), a conventional canola oil (6·2 % SFA/63·1 % MUFA) and a high-oleic acid canola oil (5·8 % SFA/74·7 % MUFA). No differences in fasting blood glucose were observed following the consumption of the dietary oils. However, when stratified by SCD genotypes, significant SNP-by-treatment interactions on blood glucose response were found with additive models for rs1502593 (P = 0·01), rs3071 (P = 0·02) and rs522951 (P = 0·03). The interaction for rs3071 remained significant (P = 0·005) when analysed with a recessive model, where individuals carrying the CC genotype showed an increase (0·14 (sem 0·09) mmol/l) in blood glucose levels with the Control oil diet, but reductions in blood glucose with both MUFA oil diets. Individuals carrying the AA and AC genotypes experienced reductions in blood glucose in response to all three oils. These findings identify a potential new target for personalised nutrition approaches aimed at improving glycaemic control.

Thermal processing not only disrupts cell membranes and cell walls, but also cleaves covalent bonds releasing low molecular phenolic. This study examined the impact of various heat treatments (100, 140, and 160°C) on the composition of phenolic acids and antioxidant activities in extracts obtained from defatted brewers spent grain (BSG) meal. Heating BSG at 160°C resulted in a 2-fold increase in total phenolic content [TPC, 172.98 ± 7.3 mg Gallic acid equivalent (GAE)/100 g defatted meal] and total flavonoid content [TFC, 16.15 ± 2.22 catechin equivalents (CE)/100 g defatted meal] compared to the untreated BSG extracts. The antioxidant activities of treated BSG extracts, determined by radical scavenging and ferric reducing antioxidant power (FRAP) were significantly (p < 0.5) higher than the corresponding untreated BSG extracts. Eleven phenolic acids were identified and quantified in BSG extracts by Ultra Performance Liquid Chromatography with Photodiode Array (UPLC-PDA). The amounts varied significantly (p < 0.05) depending on the degree of toasting BSG was subjected to. Chlorogenic acid, an ester of caffeic and quinic acid was the predominant phenolic acid present in all fractions. Significant (p < 0.05) increases in TPC, TFC, individual phenolic acids and antioxidant activity were observed in BSG extracts exposed to increasing oven temperatures. These results confirm the ability of heat processing to release bioactive phenolic from their bound forms thereby enhancing the phenolic acids and the digestibility of BSG meal in the intestinal tract.

Phenolic compounds in beer are derived from grains and hops during processing and production. Thus, the total phenolic content (TPC) of beer extracts, from selected Canadian and foreign commercial, and laboratory produced beers were examined and ranged from 3.72 ± 0.23 to 13.73 ± 0.49 mg GAE/100 mL beer. The total flavonoid content (TFC) of the same beer extracts varied from 0.82 ± 0.02 to 5.28 ± 0.04 mg CE/100 mL beer. Of particular interest were the hydroxycinnamic acids (HCAs) and derivatives in beer extracts because of their antioxidant properties. Main HCAs and a key derivative of ferulic acid, 4-vinylguaiacol (4-VG), were identified in beer extracts by high performance liquid chromatography (HPLC). Significant variations in TPC, TFC, HCAs and 4-VG content were observed among the beer extracts. Antioxidant activities of beer extracts, determined by radical scavenging and ferric-reducing assays, were positively correlated (p < 0.05) with TPC, TFC, and 4-VG but observed negative correlation with individual HCAs (p > 0.05). The positive correlation of 4-VG with ABTS radical cation scavenging and reducing power assays is reported first time. These findings suggest that phenolic compounds and a key derivative of the hydroxycinnamic acid, 4-VG, may have the protective role in the beer as an antioxidant.

The majority of the bioactive compounds present in canola seeds are phenolic compounds. They are present as aglycones, glycosides, monomers, polymerized structures, or as free or matrix-bound compounds. Conventional methods of extraction utilize an excessive amount of organic solvents and require high energy input. Current environmental issues associated with organic solvent disposal demand alternative methods for the extraction of phenolic compounds that are environmentally friendly and energy-efficient. Green technology has identified a number of alternative methods suitable for the extraction of phenolic compounds. This chapter will review those alternative green methods suitable for the extraction of phenolic compounds from canola and rapeseed. Such methods include ultrasonic extraction, microwave extraction, accelerated solvent extraction, subcritical water extraction, and supercritical fluid extraction.

Background: Different fatty acids (FAs) can vary in their obesogenic effect, and genetic makeup can contribute to fat deposition in response to dietary FA composition. However, the antiobesogenic effects of the interactions between dietary MUFAs and genetics have scarcely been tested in intervention studies. Objective: We evaluated the overall (primary outcome) and genetically modulated (secondary outcome) response in body weight and fat mass to different levels of MUFA consumption. Methods: In the Canola Oil Multicenter Intervention Trial II, a randomized, crossover, isocaloric, controlled-feeding multicenter trial, 44 men and 71 women with a mean age of 44 y and an increased waist circumference (men similar to 108 cm and women similar to 102 cm) consumed each of 3 oils for 6 wk, separated by four 12-wk washout periods. Oils included 2 high-MUFA oils-conventional canola and high-oleic canola (< 7% SFAs, > 65% MUFAs)-and 1 low-MUFA/high-SFA oil blend (40.2% SFAs, 22.0% MUFAs). Body fatwas measured using DXA. Five candidate single-nucleotide polymorphisms (SNPs) were genotyped using qualitative PCR. Data were analyzed using a repeated measures mixed model. Results: No significant differences were observed in adiposity measures following the consumption of either highMUFA diet compared with the low-MUFA/high-SFA treatment. However, when stratified by genotype, 3 SNPs within lipoprotein lipase (LPL), adiponectin, and apoE genes influenced, separately, fat mass changes in response to treatment (n = 101). Mainly, the LPL rs13702-CC genotype was associated with lower visceral fat (high-MUFA: -216.2 +/- 58.6 g; low-MUFA: 17.2 +/- 81.1 g; P = 0.017) and android fat mass (high-MUFA: -267.3 +/- 76.4 g; low-MUFA: -21.7 +/- 102.2 g; P = 0.037) following average consumption of the 2 high-MUFA diets. Conclusions: Common variants in LPL, adiponectin, and apoE genes modulated body fat mass response to dietary MUFAs in an isocaloric diet in adults with abdominal obesity. These findings might eventually help in developing personalized dietary recommendations for weight control.

IntroductionLifestyle factors, such as diet, physical activity and sleep, are associated with the development of many chronic diseases. The objective of The Manitoba Personalized Lifestyle Research study is to understand how these lifestyle factors interact with each other and with other factors, such as an individual’s genetics and gut microbiome, to influence health.MethodsAn observational study of adults, with extensive phenotyping by objective health and lifestyle assessments, and retrospective assessment of early life experiences, with retrospective and prospective utilisation of secondary data from administrative health records.Study populationA planned non-random convenience sample of 840 Manitobans aged 30–46 recruited from the general population, stratified by sex (equal men and women), body mass index (BMI; 60% of participants with a BMI>25 kg/m2) and geography (25% from rural areas). These stratifications were selected based on Manitoba demographics.MeasurementsLifestyle factors assessed will include dietary pattern, physical activity, cardiovascular fitness, and sleep. Factors such as medical history, socioeconomic status, alcohol and tobacco consumption, cognition, stress, anxiety, and early life experiences will also be documented. A maternal survey will be performed. Body composition and bone density will be measured by dual energy X-ray absorptiometry. Blood pressure, pulse wave velocity, and augmentation index will be measured on two consecutive days. Chronic disease risk biomarkers will be measured in blood and urine samples. DNA will be extracted for genetic analysis. A faecal sample will be collected for microbiome analysis. Participants may provide their Manitoba personal health information number to link their study data with administrative health records.Ethics and disseminationEthics approval has been obtained from the University of Manitoba Health Research Ethics Board (protocol # HS18951; 05/01/2016). Data analysis, release of results and publication of manuscripts are scheduled to start in early 2019. Additional information atwww.TMPLR.ca.Trial registration numberNCT03674957; Pre-results.

Background: By-products of canola oil production are currently discarded or low value commodities.This study investigates if these by-products are a significant source of extractable dietary phenols which could exhibit cellular antioxidants activity.Methods: Endogenous phenolic compounds obtained from canola oil deodistillates and canola meal using different extraction techniques were identified and examined for their in vitro antioxidant activities in "test tube" and cellular assays.Results: Sinapine was the predominant phenolic in the canola meal crude extract, while canolol was the only significant phenol in the accelerated solvent extract of canola meal.The deodistillate did not have canolol or sinapine present, but contained high molecular weight phenols of unknown identity.The "test tube" antioxidant assays indicated that canola meal crude extract and sinapic acid both exhibited stronger antioxidant potentials compared to the other extracts.A dose dependent cyto-protection effect was observed under oxidative challenge by H 2 O 2 , when cells were incubated with the canola meal accelerated solvent extract, deodistillate extract and sinapic acid.Conclusions: This study demonstrates that canola by-products can be the sources of health promoting phenols, for possible formulation into value added nutraceuticals.

Associations of individual single nucleotide polymorphisms (SNPs) in the human genes encoding for the ascorbic acid transporters SLC23A1 and SLC23A2 were reviewed. SNPs in both genes modify circulating ascorbic concentrations, the risks for various cancers, cardiovascular outcomes, inflammatory bowel disease, and ocular diseases. Disease associations are inconsistent and impact sizes are generally moderate. The impact sizes of SNPs on circulating ascorbic acid concentrations are moderate and reported changes did not alter the populations' vitamin C status. Associations for the SLC23A1 and SLC23A2 genes are currently poorly validated, limiting their utility as diagnostic or predictive biomarkers.

Existing evidence on the influence of genetic architecture on serum cholesterol responsiveness to dietary interventions focuses on individual single nucleotide polymorphisms and single nutrients. We associated the combination of ABCG5 rs6720173-C, CYP7A1 rs3808607-TT, and DHCR7 rs760241-GG genotypes with lower low-density lipoprotein cholesterol concentrations relative to the combination of rs6720173-GG, rs3808607-G, and rs760241-A genotypes (–0.37 ± 0.12 (n = 9) vs. +0.38 ± 0.14 mmol/L (n = 7), p = 0.0016) following a blended dairy (3 servings/day for 4 weeks) intervention.

Whole grain consumption is associated with reduced risk of type 2 diabetes, and the underlying mechanism might be related to the actions of polyphenols. Dietary polyphenols contribute to low glycemic indices through inhibition of intestinal glucose transport proteins. This study has two objectives: (1) to evaluate how the contents of phenolic acids in wheat vary by genetic background and growth condition and (2) to evaluate how these changes translate into physiologic relevance by investigating cellular glucose transporter inhibitions. Phenolic acids were extracted from wheat varieties grown at different locations over two crop years. The degree of inhibition of glucose uptake into human Caco-2E cells was determined. Free and bound phenolic acid extracts of all wheat genotypes inhibited glucose uptake. Degree of glucose uptake inhibitions positively correlated with the contents of free and bound phenolic acids, and the correlation coefficients were R2=0.91 and R2=0.89, respectively. Genotype and environment influenced the content of free and bound phenolic acids which linearly translated to the degree of glucose uptake inhibition in a model of intestinal absorption (P < 0.05). Results of this work mechanistically support the hypothesis that dietary phenols positively influence the glycemic index and therefore the health properties of whole grain consumption.

The effects of ultrasonic bleaching on chlorophyll reduction and oxidative stability of cold‐pressed Cannabis sativa (hemp) seed oil is investigated using three different clays, Sepiolite, activated Bentonite, and an industrial clay. The chlorophyll content is significantly reduced (p < 0.05) from 56.3 μg g−1 in the untreated oil to 14.8 μg kg−1, 9.9 μg kg−1, and 7.8 μg kg−1 in oils treated with Sepiolite, activated Bentonite, and industrial clay, respectively. Oxidative stability is enhanced in all ultrasonically‐treated oils by a significant (p < 0.05) reduction in Peroxide Value (PV) with the highest reduction (97%) observed in the presence of industrial clay (PV = 0.11 mEq kg−1). A corresponding reduction in Conjugated Dienes (CD) is also found in the treated samples ranging from 0.073–0.095% compared to 0.102% for the control. During accelerated storage at 60 °C, increase in PVs and CDs are significantly (p < 0.05) slower for the ultrasonically treated oils in the presence of clays compared to the control. Accelerated storage at 40 °C shows that the PV is greater in hempseed oil bleached with Sepiolite clay compared to a combination of Sepiolite and ultrasonic bleaching. Based on these results it is evident that the rapid removal of chlorophyll from cold pressed hempseed oil is enhanced by a combination of bleaching clay and ultrasonic treatment.Practical Applications: Ultrasonic treatment of cold pressed hempseed oil combined with bleaching clays proves very effective in reducing chlorophyll content. The method is not only rapid and clean but requires significantly less bleaching clay. Hempseed oil treated in this way exhibits greater oxidative stability making it more attractive for industrial and consumer use. The results of ultrasonic bleaching suggest its potential for prolonging the shelf‐life. Utilizing the ultrasonic bleaching technique as an alternative to conventional bleaching would be beneficial to the edible oil industry.The results of ultrasonic bleaching prove effectiveness in reducing chlorophyll content in cold‐pressed hempseed oil and suggest its potential for prolonging the shelf‐life.

The effect of different solvents (aqueous methanol [70%, v/v], aqueous acetone [80%, v/v], and a solvent mixture [MA] of aqueous methanol [70%, v/v] and aqueous acetone [70%, v/v] in a ratio of 1:1 [v/v]), preheating temperatures (140, 160, and 180 degrees C), and times of exposure (5, 15, and 30 min) on the ultrasonic extraction of the main phenolic compounds from hempseed cake (Cannabis sativa) was investigated. A simplified new high-performance liquid chromatography (HPLC) method was developed to identify and quantify the main phenolics (namely, N-transcaffeoyltyramine and cannabisin B) in the extracts. Two other main compounds, numbered 3 and 4, were also detected. The results showed that the nature of the extracting solvent had a significant (P < 0.05) impact on the ultrasonic extraction of phenolic compounds. The acetone extracts exhibited the highest total phenolic content (TPC), followed by MA and methanol. The preheating temperature and time of exposure enhanced the TPC for all solvents examined. The main phenolics, N-trans-caffeoyltyramine, cannabisin B, and compound 3, were positively affected by the temperature and time of exposure, irrespective of the solvents used. In sharp contrast, compound 4 appeared to be thermally sensitive: increasing preheating time and temperature decreased the yields of this compound. This study demonstrated that acetone was the most effective extracting solvent and that preheating enhanced the yield of the main phenolics.

Background: Crohn´s disease (CD) and ulcerative colitis (UC) are two distinct manifestations of inflammatory bowel disease (IBD).Polymorphisms in the SLC22A4 and SLC22A5 genes were associated within the IBD5 locus, but their contribution to the pathology remains unclear.Objective: This study investigated the association to IBD of common and rare variations within the SLC22A4 and SLC22A5 genes in the Manitoban IBD cohort.Design: DNA samples from 160 CD patients, 149 UC patients and 142 age and gender matched healthy controls were genotyped for selected single nucleotide polymorphisms (SNPs) tagging both genes. Results:The SLC22A5 genotypes rs11739135-CC and rs17622208-AA associated with increased susceptibility for CD (OR=7.84,95% CI 2.84-21.6,p=0.000;OR=2.26, 95% CI 1.14-4.44,p=0.019, respectively).Moreover, rs11739135-CC homozygosity was associated with UC (OR=4.18,95% CI 1. 48-11.78,p=0.007).None of the common polymorphisms tested in SLC22A4 were associated with either CD or UC.Two rarer genotypes in SLC22A4, rs11568500-A and rs11568510-G, were not detected. Conclusion:Variations in the proximal part of the SLC22A5 gene associated with IBD distinct from other variations in the IBD5 locus, including those of SLC22A4.Therefore, disturbed carnitine transport might be involved in IBD etiology in a small percentage of individuals.