Denis De Keukeleire

Withaferin a strongly elicits IkappaB kinase beta hyperphosphorylation concomitant with potent inhibition of its kinase activity

The transcription factor NFκB plays a critical role in normal and pathophysiological immune responses. Therefore, NFκB and the signaling pathways that regulate its activation have become a major focus of drug development programs. Withania somnifera (WS) is a medicinal plant that is widely used in Palestine for the treatment of various inflammatory disorders. In this study we show that the leave extract of WS, as well as its major constituent withaferin A (WA), potently inhibits NFκB activation by preventing the tumor necrosis factor-induced activation of IκB kinase β via a thioalkylation-sensitive redox mechanism, whereas other WS-derived steroidal lactones, such as withanolide A and 12-deoxywithastramonolide, are far less effective. To our knowledge, this is the first communication of IκB kinase β inhibition by a plant-derived inhibitor, coinciding with MEK1/ERK-dependent Ser-181 hyperphosphorylation. This prevents IκB phosphorylation and degradation, which subsequently blocks NFκB translocation, NFκB/DNA binding, and gene transcription. Taken together, our results indicate that pure WA or WA-enriched WS extracts can be considered as a novel class of NFκB inhibitors, which hold promise as novel anti-inflammatory agents for treatment of various inflammatory disorders and/or cancer.

Hop (Humulus lupulus)-derived bitter acids as multipotent bioactive compounds.

Hop acids, a family of bitter compounds derived from the hop plant (Humulus lupulus), have been reported to exert a wide range of effects, both in vitro and in vivo. They exhibit potential anticancer activity by inhibiting cell proliferation and angiogenesis, by inducing apoptosis, and by increasing the expression of cytochrome P450 detoxification enzymes. Furthermore, hop bitter acids are effective against inflammatory and metabolic disorders, which makes them challenging candidates for the treatment of diabetes mellitus, cardiovascular diseases, and metabolic syndrome. This review summarizes the current knowledge on hop bitter acids, including both phytochemical aspects, as well as the biological and pharmacological properties of these compounds.

Fundamentals of beer and hop chemistry

Beer brewing is an intricate process encompassing mixing and further elaboration of four essential raw materials, including barley malt, brewing water, hops and yeast. Particularly hops determine to a great extent typical beer qualities such as bitter taste, hoppy flavour, and foam stability. Conversely, hop-derived bitter acids account for an offending lightstruck flavour, which is formed on exposure of beer to light. These various processes are presented in detail, while due emphasis is placed on state-of-the-art hop technology, which provides brewers with efficient means to control bitterness, foam, and light-stability thereby allowing for the production of beers with consistent quality.

Quantitation of soy-derived phytoestrogens in human breast tissue and biological fluids by high-performance liquid chromatography.

A new and reliable HPLC method for the quantitation of daidzein, equol, and genistein in human breast tissue has been developed. The method was applied to biopsies from women undergoing breast reductions, who, prior to surgery, had ingested either a soy isoflavone preparation or a placebo tablet. The results were compared with data collected for urine and serum of the same subjects using standard methods. The limits of detection in the breast tissue homogenate were 24.7 nmol/l for daidzein, 148.0 nmol/l for equol, and 28.4 nmol/l for genistein (S/N of 3). The chromatographic limits of quantitation were 62.5 nmol/l for daidzein and genistein, and 125.0 nmol/l for equol, for which the accuracies were 86.0%, 83.6%, and 81.8%, respectively. The coefficients of variation of these measurements were all below 20% (11.1% for daidzein, 16.4% for genistein, and 13.2% for equol). The sample preparation comprised a concentration step and the absolute limits of quantitation were, therefore, 4.7 nmol/l, 18.8 nmol/l, and 0.94 nmol/l for daidzein and genistein, and 9.4 nmol/l, 37.5 nmol/l, and 1.9 nmol/l for equol in urine, serum, and breast tissue homogenate, respectively. Recoveries were between 70% (+/-5.6%) in breast tissue homogenate and 100% (+/-14.1%) in urine and serum for all three compounds. Equol (less than 1 micromol/l homogenate) was found to be the predominant phytoestrogen in breast tissue and its concentrations exceeded those in serum. The concentrations of phytoestrogens were at least 100-fold higher in urine than in serum and breast tissue.

Antiinvasive effect of xanthohumol, a prenylated chalcone present in hops (Humulus lupulus L.) and beer

The female inflorescences of the hop plant (Humulus lupulus L.) are essential during brewing to add taste and flavor to beer and to stabilize beer foam. Xanthohumol, the main prenylated chalcone in hops, was investigated for its antiinvasive activity on human breast cancer cell lines (MCF‐7 and T47‐D) in vitro. Xanthohumol was able to inhibit the invasion of MCF‐7/6 cells at 5 μM in the chick heart invasion assay and of T47‐D cells in the collagen invasion assay. Xanthohumol inhibited growth of MCF‐7/6 and T47‐D cells, but not of chick heart cells. Moreover, it induced apoptosis of these tumor cells as demonstrated by the cleavage of nuclear PARP after 48 hr treatment. To probe the mechanism of the antiinvasive effect of xanthohumol, involvement of the E‐cadherin/catenin invasion‐suppressor complex was investigated. An aggregation assay demonstrated stimulation of aggregation of MCF‐7/6 cells in the presence of 5 μM xanthohumol and this could be completely inhibited by an antibody against E‐cadherin. Xanthohumol upregulates the function of the E‐cadherin/catenin complex and inhibits invasion in vitro, indicating a possible role as an antiinvasive agent in vivo as well.

Qualitative and quantitative high-performance liquid chromatographic analysis of aldehydes in Brazilian sugar cane spirits and other distilled alcoholic beverages

A study is presented on the high-performance liquid chromatographic analysis of eighteen aldehydes in Brazilian sugar cane spirits and other international brandies. The aldehydes were separated by reversed-phase high-performance liquid chromatography as 2,4-dinitrophenylhydrazones (DNPHs). A very good chromatographic separation was achieved for eighteen different aldehyde-DNPHs. The proposed methodology is quite simple and not very time-consuming. Ten aldehydes were identified in 75 beverages and quantified using the external standard method with UV detection at 365 nm. A detailed knowledge of the aldehyde content should significantly contribute to improving the quality control of distilled spirits.

Green coconut shells applied as adsorbent for removal of toxic metal ions using fixed-bed column technology

This study applies green coconut shells as adsorbent for the removal of toxic metal ions from aqueous effluents using column adsorption. The results show that a flow rate of 2 mL/min and a bed height of 10 cm are most feasible. Furthermore, larger amounts of effluent can be treated for removal of single ions. The breakthrough curves for multiple elements gave the order of adsorption capacity: Cu(+2) > Pb(+2) > Cd(+2) > Zn(+2) > Ni(+2). Real samples arising from the electroplating industry can be efficiently handled.

Chemiluminescence-based detection: principles and analytical applications in flowing streams and in immunoassays

The present paper provides the principles of chemiluminescence (CL) and its powerful applications in analytical chemistry, mainly in the area of flow injection analysis, column liquid chromatographic and capillary electrophoretic separating systems, and its potential in immunoassays. CL is light produced by a chemical reaction. The most common advantages of chemiluminescent reactions are the relatively simple instrumentation required, the very low detection limits and wide dynamic ranges, which have contributed to the interest of CL detection in flow injection analysis, high performance liquid chromatography, including miniaturized systems, and, most recently, the exploding area of capillary electrophoresis. The latter powerful microanalytical separation technique offers high numbers of theoretical plates and relatively short analysis times requiring only small sample volumes, the migrating system comprising aqueous buffer solutions. In non-isotopic immunoassays, covering a great variety of applications in human and veterinary medicine, forensic medicine, agriculture and food industry, the radioisotope is replaced by a fluorescence or chemiluminescent label. The use of CL as a detection principle permits quantitative determination of various compounds at low concentrations. Disadvantages of the CL-based technique may include lack of sufficient selectivity and sensitivity to various physicochemical factors.

Disposition of soy isoflavones in normal human breast tissue

BACKGROUND Despite decades of research on the relation between soy and breast cancer, questions regarding the absorption, metabolism, and distribution of isoflavones in breast tissue largely remain unanswered. OBJECTIVE We evaluated the potential health effects of isoflavone consumption on normal breast tissue; isoflavone concentrations, metabolites, and biodistribution were investigated and compared with 17beta-estradiol exposure. DESIGN In this dietary intervention study, healthy women were randomly allocated to a soy milk (n = 11; 16.98-mg genistein and 5.40-mg daidzein aglycone equivalents per dose), soy supplement (n = 10; 5.27-mg genistein and 17.56-mg daidzein aglycone equivalents per dose), or control (n = 10) group. After a run-in period > or = 4 d, 3 doses of soy milk or soy supplements were taken daily for 5 d before an esthetic breast reduction. Blood and breast biopsies were collected during surgery and analyzed with liquid chromatography-tandem mass spectrometry. RESULTS After soy administration, genistein and total daidzein concentrations, which were expressed as aglycone equivalents, ranged from 135.1 to 2831 nmol/L and 105.1 to 1397 nmol/L, respectively, in hydrolyzed serum and from 92.33 to 493.8 pmol/g and 22.15 to 770.8 pmol/g, respectively, in hydrolyzed breast tissue. The major metabolites identified in nonhydrolyzed samples were genistein-7-O-glucuronide and daidzein-7-O-glucuronide, with an overall glucuronidation of 98%. Total isoflavones showed a breast adipose/glandular tissue distribution of 40:60, and their mean (+/-SEM) derived 17beta-estradiol equivalents toward estrogen receptor beta were 21 +/- 4-fold and 40 +/- 10-fold higher than the 17beta-estradiol concentrations in adipose (0.283 +/- 0.089 pmol/g, P < 0.001) and glandular (0.246 +/- 0.091 pmol/g, P = 0.001) fractions, respectively. CONCLUSION After intake of soy milk and soy supplements, isoflavones reach exposure levels in breast tissue at which potential health effects may occur.

Structure-activity relationship of short-chain sphingoid bases as inhibitors of sphingosine kinase.

Short-chain sphinganine analogues 8, 9, 18, and 19, as well as 3-fluoro-sphingosine analogues 25 and 26 were synthesized. Their potential as sphingosine kinase inhibitors was investigated, in combination with previously synthesized sphingosine and fluorinated sphinganine analogues.