Khasianine Affects the Expression of Sugar-Sensitive Proteins in Pancreatic Cancer Cells, Which Are Altered in Data from the Rat Model and Patients.

Pancreatic ductal adenocarcinoma (PDAC) is a deadly malignancy with no effective treatment, particularly in the advanced stage. This study explored the antiproliferative activity of khasianine against pancreatic cancer cell lines of human (Suit2-007) and rat (ASML) origin. Khasianine was purified from Solanum incanum fruits by silica gel column chromatography and analyzed by LC-MS and NMR spectroscopy. Its effect in pancreatic cancer cells was evaluated by cell proliferation assay, chip array and mass spectrometry. Proteins showing sensitivity to sugars, i.e. sugar-sensitive lactosyl-Sepharose binding proteins (LSBPs), were isolated from Suit2-007 cells by competitive affinity chromatography. The eluted fractions included galactose-, glucose-, rhamnose- and lactose-sensitive LSBPs. The resulting data were analyzed by Chipster, Ingenuity Pathway Analysis (IPA) and GraphPad Prism. Khasianine inhibited proliferation of Suit2-007 and ASML cells with IC50 values of 50 and 54 µg/mL, respectively. By comparative analysis, khasianine downregulated lactose-sensitive LSBPs the most (126%) and glucose-sensitive LSBPs the least (85%). Rhamnose-sensitive LSBPs overlapped significantly with lactose-sensitive LSBPs and were the most upregulated in data from patients (23%) and a pancreatic cancer rat model (11.5%). From IPA, the Ras homolog family member A (RhoA) emerged as one of the most activated signaling pathways involving rhamnose-sensitive LSBPs. Khasianine altered the mRNA expression of sugar-sensitive LSBPs, some of which were modulated in data from patients and the rat model. The antiproliferative effect of khasianine in pancreatic cancer cells and the downregulation of rhamnose-sensitive proteins underscore the potential of khasianine in treating pancreatic cancer.

Ethanol extracts of mango seeds added to the diet of pigs increases antioxidant capacity of processed pork.

Synthetic antioxidants (e.g.butylhydroxytoluene, BHT) are routinely used for to restrict oxidative processes of meat products, but they are implicated as harmful to the health of humans. Therefore natural alternatives, such as plant antioxidants, have been sought as replacements. Plant antioxidants when added to the diet can be incorporated into meat and reduce the need for the addition of synthetic antioxidants during processing. The objective of this study was to evaluate the effects of ethanol extracts of mango seeds (EEMS) in the diet of pigs on qualitative parameters and total antioxidant capacity of mortadella produced from these animals. Thirty-two pigs with an average 60 days of age were distributed among four treatments: control=no antioxidant; BHT=200ppm BHT; EEMS200=200ppm of EEMS and EEMS400=400ppm of EEMS. At 145 days of age the animals were slaughtered and loin was removed for the preparation of mortadella, which was analyzed during 90 days of storage at 4°C. A higher content of polyphenolic compounds and, total antioxidant capacity in mortadellas processed with meat of animals which consumed the EEMS400 ration after 60 and 90 days of storage was observed. EEMS polyphenolic antioxidants incorporated into pork through the diet results in an increase of total antioxidant capacity in the processed product.

The concentration of polyphenolic compounds and trace elements in the Coffea arabica leaves: Potential chemometric pattern recognition of coffee leaf rust resistance.

Coffee (Coffea arabica L.) is an important commodity, involving about 500 million people from the cultivation of the coffee trees to final consumption of infusions of the ground roasted coffee beans. In contrast to a considerable amount of research performed on green coffee beans, there are relatively few studies regarding the chemical constituents of coffee leaves. Hemileia vastatrix is a parasite, specific to coffee plants and causes coffee leaf rust, which is a very destructive disease. Some coffee plants have natural resistance which is mainly linked to a gene and specific host resistance response. An increase in flavonoid production may be related to fungal disease resistance, with the levels and flavonoid types being an early physiological response to rust infection. Trace inorganic elements can be related to many roles in the defense response of higher plants and can be used as a biomarker for some diseases. To address this, coffee leaves from 16 different cultivars of Coffea arabica were harvested from Minas Gerais, Brazil (susceptible and resistant to rust) and their polyphenolic compounds were extracted using the QuEChERS technique and quantitated by HPLC-ESI-MS. The same leaves were decomposed using an acid mixture in a block digester and the content of Al, Cu, Mg, Mn, Ni, Sn and Zn was quantitated by ICP-OES. Principal component analysis (PCA) was applied and we could establish a relation between polyphenolic and trace element concentration in the leaves with resistance to rust infection. On this basis in this preliminary study we were able to separate the resistant from the susceptible cultivars. The main compounds responsible for this differentiation were the content of chlorogenic acid and magnesium in the leaves. The content of polyphenolic compounds was lower in susceptible cultivars and a diametric effect was observed between Mn and Mg concentrations. This study shows potential for the discrimination of resistant and susceptible coffee trees based on the analyses of both trace element and polyphenolic concentration.

Expression Patterns of Xenobiotic-Metabolizing Enzymes in Tumor and Adjacent Normal Mucosa Tissues among Patients with Colorectal Cancer: The ColoCare Study.

BACKGROUND: Xenobiotic-metabolizing enzymes (XME) play a critical role in the activation and detoxification of several carcinogens. However, the role of XMEs in colorectal carcinogenesis is unclear. METHODS: We investigated the expression of XMEs in human colorectal tissues among patients with stage I-IV colorectal cancer (n = 71) from the ColoCare Study. Transcriptomic profiling using paired colorectal tumor and adjacent normal mucosa tissues of XMEs (GSTM1, GSTA1, UGT1A8, UGT1A10, CYP3A4, CYP2C9, GSTP1, and CYP2W1) by RNA microarray was compared using Wilcoxon rank-sum tests. We assessed associations between clinicopathologic, dietary, and lifestyle factors and XME expression with linear regression models. RESULTS: GSTM1, GSTA1, UGT1A8, UGT1A10, and CYP3A4 were all statistically significantly downregulated in colorectal tumor relative to normal mucosa tissues (all P ≤ 0.03). Women had significantly higher expression of GSTM1 in normal tissues compared with men (ß = 0.37, P = 0.02). By tumor site, CYP2C9 expression was lower in normal mucosa among patients with rectal cancer versus colon cancer cases (ß = -0.21, P = 0.0005). Smokers demonstrated higher CYP2C9 expression levels in normal mucosa (ß = 0.17, P = 0.02) when compared with nonsmokers. Individuals who used NSAIDs had higher GSTP1 tumor expression compared with non-NSAID users (ß = 0.17, P = 0.03). Higher consumption of cooked vegetables (>1×/week) was associated with higher CYP3A4 expression in colorectal tumor tissues (ß = 0.14, P = 0.007). CONCLUSIONS: XMEs have lower expression in colorectal tumor relative to normal mucosa tissues and may modify colorectal carcinogenesis via associations with clinicopathologic, lifestyle, and dietary factors. IMPACT: Better understanding into the role of drug-metabolizing enzymes in colorectal cancer may reveal biological differences that contribute to cancer development, as well as treatment response, leading to clinical implications in colorectal cancer prevention and management.

Study of Antihypertensive Activity of Anvillea radiata in L-Name-Induced Hypertensive Rats and HPLC-ESI-MS Analysis.

OBJECTIVE: This study aimed to evaluate the effect of the aqueous extract of Anvillea radiate (A. radiata) aerial parts (AEAR) on arterial blood pressure in normotensive and hypertensive rats. METHODS: The effect of the acute and sub-chronic administration of AEAR on the following blood pressure parameters: systolic blood pressure (SBP), mean blood pressure (MBP), diastolic blood pressure (DBP), and heart rate (HR) was evaluated in normotensive and L-NAME induced hypertensive rats. In the second experiment, the vasorelaxant effect of AEAR was assessed in isolated aortic rings from rats with functional endothelium pre-contracted with epinephrine (EP) or KCl, and six antagonists/ inhibitors were used to explore the mechanisms of action involved in the vasorelaxant effect. In order to determine the phytochemical contents of Anvillea radiata, HPLC-ESI-MS analysis was conducted. RESULTS: Daily oral administration of AEAR (100 mg/kg) provoked a significant decrease in SBP, MBP, and DBP without affecting HR in hypertensive rats. In addition, AEAR (0.08-0.64 mg/ml) revealed a vasorelaxant effect in thoracic aortic rings pre-contracted by EP (10 µM) or KCl (80 mM). This effect was reduced in the presence of Nifedipine, L-Name or Methylene blue. The polyphenolic compounds of AEAR were determined. CONCLUSION: This study revealed that AEAR possesses a potent antihypertensive activity and its vasorelaxant activity seems to be mediated through Ca2+ channels, direct nitric oxide (NO), and NO/cGMP pathways. Chlorogenic acid and caffeic acid identified in A. radiata could be at least partially responsible for the antihypertensive activity of this extract.

Chromium (VI) remediation in aqueous solution by waste products (peel and seed) of mango (Mangifera indica L.) cultivars.

The surface group characteristics of mango cultivar peels and seeds were evaluated by infrared spectra, PZC, and functional group composition. The adsorption/reduction of chromium (VI) in aqueous solutions was investigated by varying pH, contact time, initial Cr(VI) concentration, and adsorbent amount. The results show that both peel and seed powders of the mango cultivars showed significant adsorption/reduction capacity for Cr(VI) and that the desorption process obeys pseudo-second-order kinetics. Optimal adsorption occurred at pH 1.0, using a Cr(VI) concentration of 100 mg/L. On average, at pH 1.0, and a concentration of 3 g/L, the maximum adsorption/reduction capacity of Cr(VI) was 83% (peels 76%, seeds 90%). Of the mango powders tested, the most efficient were Tommy seed (100%) and Coite peel (98%) followed by Coite seed (96%) and Tommy peel powders (95%). The adsorption/reduction of Cr(VI) was complete (100%) by the mango seed, in comparison to the peel powders (97%) after 180 min. The data indicates that mango waste products, such as seed and peel powders, are both excellent candidates for the remediation of Cr(VI) from aqueous systems and due to the higher concentration of gallates and galloyl glucosides, the mango seed powders should be the powders of choice for future remediation projects.

Nutraceutical compounds: Echinoids, flavonoids, xanthones and caffeine identified and quantitated in the leaves of Coffea arabica trees from three regions of Brazil.

There are relatively few studies concerning the use of coffee leaves for medicinal purposes and the composition of secondary plant substances. Therefore, we identified and quantitated polyphenolic compounds along with caffeine present in methanol extracts of Coffea arabica leaves from three different regions of Brazil (Ceará, Minas Gerais and São Paulo) by HPLC-ESI-MS. In addition, correlations between polyphenolic content of the coffee leaves and antioxidant assays DPPH, FRAP and ORAC were evaluated. Fifteen compounds belonging to three classes of polyphenols (xanthones, chlorogenic acids and flavonoids) along with the alkaloid caffeine were detected. The mean concentration of total polyphenolic compounds in the leaves of C. arabica, harvested from three different regions of Brazil was quite variable. The highest values were detected in the coffee leaves harvested in Minas Gerais (n = 4) at 40.80(13.00) g/kg (SD), followed by coffee leaves harvested in São Paulo (n = 20) at 24.79(20.19) g/kg, and the lowest in coffee leaves harvested in Ceará (n = 11) in the Northeast of Brazil at 10.30(5.61) g/kg. The three classes of polyphenols, all showed excellent correlations in the antioxidant assays. Coffee leaf tea, appears to be an excellent functional beverage, with its high content of polyphenolic compounds, which may render positive biologic effects, when inbibed as part of the normal human diet.

Evaluation of Glucose and Lipid Lowering Activity of Arganimide A in Normal and Streptozotocin-Induced Diabetic Rats.

AIMS: Arganimide A (4,4-dihydroxy-3,3-imino-di-benzoic acid) is a compound belonging to a family of aminophenolics found in fruit of Argania spinosa. The purpose of this study was to investigate the glucose and lipid lowering activity of Arganimide A (ARG A). METHODS: The effect of a single dose and daily oral administration of Arganimide A (ARG A) on blood glucose levels and plasma lipid profile was tested in normal and streptozotocin (STZ) diabetic rats at a dose of 2 mg/kg body weight. RESULTS: Single oral administration of ARG A reduced blood glucose levels from 26.50±0.61 mmol/L to 14.27±0.73 mmol/L (p<0.0001) six hours after administration in STZ diabetic rats. Furthermore, blood glucose levels were decreased from 5.35±0.30 mmol/L to 3.57±0.17 mmol/L (p<0.0001) and from 26.50±0.61 mmol/L to 3.67±0.29 mmol/L (p<0.0001) in normal and STZ diabetic rats, respectively, after seven days of treatment. Moreover, no significant changes in body weight in normal and STZ rats were shown. According to the lipid profile, the plasma triglycerides levels were decreased significantly in diabetic rats after seven days of ARG treatment (p<0.05). Moreover, seven days of ARG A treatment decreased significantly the plasma cholesterol concentrations (p<0.001). CONCLUSION: ARG A possesses glucose and lipid-lowering activity in diabetic rats and this natural compound may be beneficial in the treatment of diabetes.

Improved Methods for the Rapid Formation and Prevention of Advanced Glycation End Products (AGEs) In Vitro by Coupling to the Hypoxanthine/Xanthine Oxidase Assay System.

Advanced glycation end products (AGEs) represent a set of molecules that contribute directly to the initiation and aggravation of diseases associated with ageing. AGEs are produced by the reaction between reducing sugars (or α-dicarbonyl compounds), proteins, and amino acid residues. Previous in vitro methods using non-enzymatic procedures described in the literature require an incubation period of 1⁻3 weeks to generate AGEs. In this study, the reaction time for the formation of AGEs (48 and 3 h) was significantly reduced by adaptation of methods previously described in the literature and coupling them to the free radical generation system termed hypoxanthine/xanthine oxidase assay. The incorporation of this assay into the experimental system accelerated the production of AGEs as a result of the formation of reactive oxygen species (ROS), as shown by increased fluorescence. The capacity of different classes of chemical compounds (aminoguanidine, chlorogenic acid, rutin, and methanol extracts of Hancornia speciosa Gomes) to inhibit protein glycation by acting as scavenging agents of α-dicarbonyl species was evaluated. Aminoguanidine and, especially, rutin identified in the leaf extracts of H. speciosa Gomes showed a high capacity to act as scavengers of reactive carbonyl species RCS-trapping, resulting in the inhibition of AGEs formation.

Dose-Response Relationship between Serum Retinol Levels and Survival in Patients with Colorectal Cancer: Results from the DACHS Study.

Current knowledge on the role of retinol in the prognosis of patients with colorectal cancer (CRC) is very limited. We investigated the association of serum retinol levels with survival outcomes in a large cohort of 2908 CRC patients from Germany. Retinol concentrations were determined in serum collected shortly after diagnosis by mass spectrometry. Associations between serum retinol levels and survival outcomes were assessed using multivariable Cox regression and dose-response analyses. The joint association of serum retinol and serum 25-hydroxyvitamin D3 (25(OH)D3) with survival outcomes was also examined. During a median follow-up of 4.8 years, 787 deaths occurred, 573 of which were due to CRC. Dose-response curves showed an inverse relationship between serum retinol levels and survival endpoints in the range of <2.4 µmol/L, but no associations at higher levels. Low (<1.2 µmol/L) versus high (≥2.4 µmol/L) serum retinol levels were associated with poorer overall survival (Hazard ratio (HR) = 1.46, 95% confidence interval (CI) = 1.19⁻1.78, P-trend = 0.0003) and CRC-specific survival (HR = 1.69, 95% CI = 1.33⁻2.15, P-trend < 0.0001). Joint presence of low serum retinol (<1.2 µmol/L) and low 25(OH)D3 (<30 nmol/L) was associated with a particularly strong decrease in overall and CRC-specific survival. Low serum retinol levels were identified as a predictor of poor survival in CRC patients, in particular when co-occurring with low serum concentrations of 25(OH)D3. The clinical implications of these findings require further investigation.

Relationship of very low serum 25-hydroxyvitamin D3 levels with long-term survival in a large cohort of colorectal cancer patients from Germany.

To investigate the association of serum 25-hydroxyvitamin D (25(OH)D3) with survival in a large prospective cohort study of colorectal cancer (CRC) patients. The study population consisted of 2,910 patients diagnosed with CRC between 2003 and 2010 who participated in the DACHS study, a multicenter study from Germany with comprehensive long-term follow-up. 25(OH)D3 was determined in serum samples collected shortly after cancer diagnosis by High Performance Liquid Chromatography-Electro Spray Ionization-Mass Spectrometry. Analyses of survival outcomes were performed using Cox regression with comprehensive adjustment for relevant confounders. The majority (59%) of CRC patients were vitamin D deficient (serum 25(OH)D3 levels <30 nmol/L). During a median follow-up of 4.8 years, 787 deaths occurred, 573 of which were due to CRC. Compared to patients in the highest 25(OH)D3 quintile (>45.20 nmol/L), those in the lowest 25(OH)D3 quintile (<11.83 nmol/L) had a strongly increased mortality. Adjusted hazard ratios (95% Confidence Interval) were 1.78 (1.39-2.27), 1.65 (1.24-2.21), 1.32 (1.03-1.71) and 1.48 (1.18-1.85) for all-cause mortality, CRC-specific mortality, recurrence-free and disease-free survival, respectively. Subgroup analyses did not show any significant effect modification across strata defined by sex, age, stage, body mass index, or the late entry. Dose-response analyses showed a strong inverse relationship between serum 25(OH)D3 levels and survival endpoints at 25(OH)D3 levels <30 nmol/L, and no association with mortality at higher 25(OH)D3 levels. Vitamin D deficiency is highly prevalent in CRC patients and a strong independent predictor of poor prognosis. The possibility of enhancing CRC prognosis by vitamin D supplementation, ideally combined with outdoor physical activity, should be evaluated by randomized controlled trials focusing on patients with vitamin D deficiency.

Bioassay-Guided Isolation and Evaluation of Antiproliferative Effects of (Z)-Ethylidene-4, 6-Dimethoxycoumaran-3-One from Pogostemon Quadrifolius (Benth.)

The aim of the study was to isolate and identify the major cytotoxic principle from plant leaves of Pogostemon quadrifolius (Benth.) and evaluate its antiproliferative potential against human cancer cells. Plant leaves were extracted sequentially with a soxhlet apparatus, using petroleum ether, chloroform and methanol solvents. Petroleum ether and chloroform extracts exhibited antiproliferative properties against Caco-2, HeLa, THP-1, MCF-7 and Jurkat E6-1cancer cell lines tested, but methanol extracts failed to exhibit such activity. The major antiproliferative principle from petroleum ether and chloroform extracts was isolated with the help of bioassay guided column chromatography. This cytotoxic compound was further analysed by UV, TLC, HPLC, LC-MS, GC-MS and NMR analyses and was identified to be novel: (Z)-ethylidene-4,6-dimethoxycoumaran-3-one (Compound 1). The half-maximal inhibitory concentrations for proliferation (IC50) exhibited by compound 1 were 19.4, 23.1, 22.1, 35.9 and 8.32 µM against Caco-2, HeLa, THP-1, MCF-7 and Jurkat E6-1 cancer cell lines, respectively. Further experiments revealed that compound 1 triggered the apoptosis mode of cell death in cancer cell lines. Thus, the present study allowed isolation and identification of a novel cytotoxic natural compound, (Z)-ethylidene-4,6-dimethoxycoumaran-3-one, from plant leaves of P. quadrifolius (Benth.). Our pre-clinical study also indicated that compound 1 is particularly active in the acute T cell leukemia cell line (Jurkat E6-1) with potential for application as a chemotherapeutic agent in the future.

Plasma 25-Hydroxyvitamin D3 Levels in Colorectal Cancer Patients and Associations with Physical Activity.

Physical activity (PA) and vitamin D are thought to affect colorectal cancer prognosis. The present study investigates associations between 25(OH)D3 and PA in prospectively followed colorectal cancer patients in the ColoCare study. At 6, 12, and 24 mo after surgery, patients donated a blood sample, wore an accelerometer for 10 consecutive days, and completed a PA questionnaire. Plasma 25-hydroxyvitamin D3 (25(OH)D3) levels were measured by high-performance liquid chromatography. We tested associations using partial correlations and multivariate linear regression analysis, adjusted for season, age, and body mass index. A total of 137 assessments of 25(OH)D3 levels and PA were conducted (58 at 6 mo, 51 at 12 mo, and 28 at 24 mo). More than 60% of the patients were vitamin D-deficient (25(OH)D3 ≤20 ng/ml), independent of study time point. At 6-mo follow-up, accelerometry-based vigorous and moderate-to-vigorous PAs were positively associated with 25(OH)D3 levels (P = 0.04; P = 0.006,). PA together with season was a significant predictor of elevated 25(OH)D3 levels. Our results suggest that the majority of colorectal cancer patients may suffer from vitamin D deficiency. Engaging in PA may be an effective approach to increase their 25(OH)D3 levels.

Comparison of the major polyphenols in mature Argan fruits from two regions of Morocco.

Previous studies have shown that Argan fruits contain a large variety of polyphenolic compounds. Recently, another class of polyphenolic compounds, namely amino phenols have been detected and identified in immature Argan fruits. The objective of this study, was to establish whether or not, these novel compounds are also present in mature Argan fruits. To this end, a comparison was made between mature fruits from two regions of Morocco. Nineteen major compounds were identified and quantitated, including amino phenols, flavonoids, and phenolic acids by chromatographic methods in mature Argan fruits from the two regions of Morocco (Essaouira and Agadir). The phenolic acids were identified as gallic acid and 3,4-dihydroxybenzoic acid; the amino phenols as Arganimide A, and argaminolics A-C, and the flavonoids as rutin pentoside, quercetin-3-O-arabinoside, quercetin glycogallate, quercetin-3-O-rhamnogalactoside, rutin, quercetin-3-O-galactoside (hyperoside), quercetin-3-O-glucoside (quercitrin), quercetin-3-O-arabinoside, quercetin glycohydroxybenzoate, quercetin glycosinapate, quercetin glycoferulate, quercetin glycocoumarate and quercetin. n=145.

Characterization and quantitation of the polyphenolic compounds detected in methanol extracts of Pistacia atlantica Desf. fruits from the Guelmim region of Morocco.

High performance liquid chromatography coupled with electrospray ionization mass spectrometry (HPLC-ESI-MS) was used for the identification of the major phenolic compounds in mature P. atlantica fruits from the Guelmim region (southeast of Morocco). In this study twenty seven polyphenolic compounds are identified and quantitated. To date, this is the most comprehensive report on the polyphenolic content of Pistacia fruits. The profiles comprise, three major polyphenolic classes, namely gallates (18.76g/kg; 63.92%), flavonoids (10.12g/kg; 34.48%) and ellagic acid derivatives (0.47g/kg; 1.60%) with a total of 29.35g/kg detected. The major gallate was pentagalloyl glucoside (5.0g/kg; 17.04% of total polyphenolics), the major flavonoid luteolin (3.18g/kg; 10.83% of total polyphenolics) and the major ellagic acid derivative ellagic acid (0.25g/kg; 0.85% of total polyphenolics). Identification of galloyl quinate, digalloyl quinates (x 2), galloyl glucoside, digalloyl glucosides (x 2), trigalloyl glucoside, tetragalloyl glucosides (x 2), pentagalloyl glucoside, 2″-O-galloyl-quercetin-3-O-galactoside, quercetin-3-O-rhamnogalactoside, quercetin-3-O-galactoside, ellagic acid diglucoside, luteolin-4'-O-glucoside, 2″-O-galloyl-luteolin-4'-O-glucoside, quercetin-3-O-glucuronide, kaempferol-3-O-glucoside, eriodictyol, apigenin, ellagic acid diglucoside, ellagic acid glucoside, methyl ellagic acid glucoside, and ellagic acid are described as phytochemical components of Pistacia fruits for the first time.

A click chemistry approach identifies target proteins of xanthohumol.

SCOPE: Many phytochemicals with beneficial pharmacological properties contain electrophilic sites, e.g. α,ß-unsaturated carbonyl (enone) groups. There is increasing evidence that many biological effects of electrophilic compounds depend on covalent conjugation to reactive protein thiols. For example, the reaction of electrophiles with cysteinyl residues of the sensor protein Keap1 activates the cell-protective Nrf2 response. Thus it is of interest to identify more generally the proteins to which small molecule electrophiles bind covalently. METHODS AND RESULTS: Here we use a Click chemistry approach to identify target proteins of the chemopreventive phytochemical xanthohumol (XN), an enone-containing chalcone from hops (Humulus lupulus L.). Using an alkynylated analog of XN (XN-alkyne), we purified covalent protein-electrophile conjugates from cell lysates. We confirm the previously described conjugation of XN to Keap1. One of the newly identified candidate target proteins is glucose-6-phosphate dehydrogenase (G6PDH). We confirm that XN attenuates intracellular G6PDH activity at low micromolar concentrations. CONCLUSION: We find support for the notion that XN modulates multiple pathways and processes by covalent modification of proteins with reactive cysteines.

Identification of polyphenolic compounds in the flesh of Argan (Morocco) fruits.

High performance liquid chromatography coupled with negative electrospray ionization (HPLC-ESI) along with fragmentation patterns generated by nano-electrospray ionization (nano-ESI-MS-MS) and NMR techniques were utilized for the identification of phenolic compounds in Argan fruits. A total of 15.4 g/kg was determined represented by catechins (39%), flavonoids (28%), procyanidins (26%), free phenolic acids (6%) and phenolic acid glycosides (1%). Twenty-one phenolic compounds were identified for the first time in Argan fruits namely III. epicatechin-(4ß→8)-catechin dimer (procyanidin B1), IV. p-coumaric acid glycoside, VI. epicatechin-(4ß→8)-epicatechin dimer (procyanidin B2), VIII. caffeic acid glycoside, XIX. epicatechin-(4ß→8)-epicatechin-(4ß→8)-epicatechin trimer (procyanidin C1), X. p-hydroxybenzaldehyde XI. ferulic acid glycoside, XII. vanillic acid, XIII. sinapic acid glycoside, XVI. p-coumaric acid, XVII. ferulic acid, XVIII. sinapic acid, XIX. rutin pentoside, XX. quercetin glycopentoside, XXI. 4,4'-dihydroxy-3,3'-imino-di-benzoic acid, XXV. quercetin-3-O-rhamnogalactoside, XXVII. quercetin glycohydroxybenzoate, XXVIII. quercetin glycocaffeate, XXIX. quercetin glycosinapate, XXX. quercetin glycoferulate and XXXI. quercetin glycocoumarate.

Changes in urinary metabolic profiles of colorectal cancer patients enrolled in a prospective cohort study (ColoCare).

Introduction: Metabolomics is a valuable tool for biomarker screening of colorectal cancer (CRC). In this study, we profiled the urinary metabolomes of patients enrolled in a prospective patient cohort (ColoCare). We aimed to describe changes in the metabolome in the longer clinical follow-up and describe initial predictors as candidate markers with possibly prognostic significance. Methods: In total, 199 urine samples from CRC patients pre-surgery (n=97), 1-8 days post-surgery (n=12) and then after 6 and 12 months (n=52 and 38, respectively) were analyzed using both GC-MS and 1H-NMR. Both datasets were analyzed separately with built in uni- and multivariate analyses of Metaboanalyst 2.0. Furthermore, adjusted linear mixed effects regression models were constructed. Results: Many concentrations of the metabolites derived from the gut microbiome were affected by CRC surgery, presumably indicating a tumor-induced shift in bacterial species. Associations of the microbial metabolites with disease stage indicate an important role of the gut microbiome in CRC.We were able to differentiate the metabolite profiles of CRC patients prior to surgery from those at any post-surgery timepoint using a multivariate model containing 20 marker metabolites (AUCROC=0.89; 95% CI:0.84-0.95). Conclusion: To the best of our knowledge, this is one of the first metabolomic studies to follow CRC patients in a prospective setting with repeated urine sampling over time. We were able to confirm markers initially identified in case-control studies and pin point metabolites which may serve as candidates for prognostic biomarkers of CRC.

Pilot walnut intervention study of urolithin bioavailability in human volunteers.

A pilot intervention study was conducted in human volunteers (n = 4) to establish the bioavailability of urolithins, which are the terminal end-products of ellagitannin metabolism by the gastrointestinal microflora. Biospecimens (blood, feces, and urine) along with urolithins purified therefrom were analyzed for their antioxidant capacity in a range of in vitro assays. Urolithin metabolites were identified and quantitated in the biospecimens by negative ion mode HPLC-ESI-MS analysis. The data in this pilot study show that the metabolism of ellagitannins in the four volunteers gave rise to a diverse profile and a highly variable concentration of urolithins in urine. The concentration of glucuronidated urolithins in blood and urine did not correlate with antioxidant capacity. However, the antioxidant capacity of urine, but not plasma biospecimens, was highly correlated with uric acid concentration. The antioxidant capacity of fecal extracts correlated positively with the concentration of urolithin D in both the DPPH and FRAP assays, but not in the ORAC assay, which was entirely consistent with the in vitro assays for pure urolithin D.

Amino phenolics from the fruit of the argan tree Argania spinosa (Skeels L.).

A new phenolic-type compound containing a nitrogenous, heterocyclic-fused ring from the fruit of the argan tree, Argania spinosa (Skeels L.), is described. This and another already known compound also isolated in the course of the work belong to an obscure and rare class of natural products, the amino phenolics.