Induction of stilbenes in grapes by UV-C: Comparison of different subspecies of Vitis

Bioactive products enriched in stilbenes are considered of potential future interest, and the main sources of stilbenes in human diet are grapes. Postharvest ultraviolet C (UV-C) treatment was used to induce stilbene biosynthesis in grapes of three varieties of Vitis vinifera sylvestris, seven of Vitis vinifera sativa, and two Hybrid Direct Producers (HDPs). Stilbenes have been identified by UPLC-DAD-TQD and quantified by HPLC-DAD, and cluster analyses have been performed to classify subspecies by their stilbene profile. After UV-C treatment, the Syrah variety reached a maximum of 25 mg kg^? 1 f.w. of total stilbenes in the 2008 vintage, and in the statistical analysis, this variety stood out from the other Vitis varieties tested. In 2008, varieties belonging to the sylvestris group and Vitis vinifera sativa Merlot also presented high stilbene production; however, the expected concentration in the HDPs was not obtained.Industrial relevanceIn this study, postharvest UV-C treatment has been applied to obtain stilbene-enriched grapes from several subspecies of Vitis. The most suitable raw material for bioactive stilbene-enriched products, such as nutraceutics and wines, has been established. This is an essential prerequisite to scaling up the process. Such stilbene-enriched products are known to have added-value as a result of the confirmed positive bioactivity of these compounds, and are gaining market acceptance.     

New Pacidamycin Antibiotics Through Precursor‐Directed Biosynthesis

Pacidamycins, mureidomycins and napsamycins are structurally related uridyl peptide antibiotics that inhibit translocase I, an as yet clinically unexploited target. This potentially important bioactivity coupled to the biosynthetically intriguing structure of pacidamycin make this natural product a fascinating subject for study. A precursor‐directed biosynthesis approach was employed in order to access new pacidamycin derivatives. Strikingly, the biosynthetic machinery exhibited highly relaxed substrate specificity with the majority of the tryptophan analogues that were administered; this resulted in the production of new pacidamycin derivatives. Remarkably, 2‐methyl‐, 7‐methyl‐, 7‐chloro‐ and 7‐bromotryptophans produced their corresponding pacidamycin analogues in larger amounts than the natural pacidamycin. Low levels or no incorporation was observed for tryptophans substituted at positions 4, 5 and 6. The ability to generate bromo‐ and chloropacidamycins opens up the possibility of further functionalising these compounds through chemical cross‐coupling in order to access a much larger family of derivatives.     

Human dietary exposure to heavy metals via the consumption of greenshell mussels (Perna canaliculus Gmelin 1791) from the Bay of Islands, northern New Zealand

Cadmium (Cd), mercury (Hg), arsenic (As), lead (Pb) and tin (Sn) concentrations were determined using ICP-MS in soft tissues (wet wt.) from whole greenshell mussels (Perna canaliculus) collected from Urapukapuka-Rawhiti Island, Opua Marina, Waitangi Bridge and Opua Wharf from the Bay of Islands, northern New Zealand (NZ). All samples had metal concentrations well below the Food Standards Australia and New Zealand (FSANZ) maximum limits and were comparable to, or less than, concentrations observed in previous NZ studies. Based on the average values detected in the current study, the concentrations of heavy metals ingested in a ‘typical diet’ containing greenshell mussels are below the provisional tolerable weekly intake (PTWI). However, Māori (indigenous people of New Zealand), Pacific Islanders and Asians consume a far greater quantity of seafood (and therefore heavy metals) than the general public of New Zealand and could potentially consume enough shellfish to exceed the PTWI for Cd (but not for Hg, As, Pb or Sn). Although our results, based on the current PTWIs, indicate no significant health risk to greenshell mussel consumers in this region, PTWIs change over time; concentrations which were thought to be safe are later found to be harmful. Additionally, differences in individual human susceptibilities to various toxins could increase the risk of harm for consumers with low tolerance to heavy metals. We suggest that a survey of the frequency, amount and species consumed by groups whose diet may be largely shellfish-based is required to enable a more comprehensive risk assessment to be made.     

Synthesis and Evaluation of 1‐(1‐(Benzo[b]thiophen‐2‐yl)cyclohexyl)piperidine (BTCP) Analogues as Inhibitors of Trypanothione Reductase

Thirty two analogues of phencyclidine were synthesised and tested as inhibitors of trypanothione reductase (TryR), a potential drug target in trypanosome and leishmania parasites. The lead compound BTCP ( 1 , 1‐(1‐benzo[b]thiophen‐2‐yl‐cyclohexyl) piperidine) was found to be a competitive inhibitor of the enzyme (K_i=1 μM ) and biologically active against bloodstream T. brucei (EC₅₀=10 μM ), but with poor selectivity against mammalian MRC5 cells (EC₅₀=29 μM ). Analogues with improved enzymatic and biological activity were obtained. The structure–activity relationships of this novel series are discussed.     

Highly Selective and Potent Human β-Secretase 2 (BACE2) Inhibitors against Type 2 Diabetes: Design,Synthesis, X-ray Structure and Structure–Activity Relationship Studies

Herein we present the design, synthesis, and biological evaluation of potent and highly selective β-secretase 2 (memapsin 1, beta-site amyloid precursor protein cleaving enzyme 2, or BACE 2) inhibitors. BACE2 has been recognized as an exciting new target for type 2 diabetes. The X-ray structure of BACE1 bound to inhibitor 2 a {N³-[(1S,2R)-1-benzyl-2-hydroxy-3-[[(1S,2S)-2-hydroxy-1-(isobutylcarbamoyl)propyl]amino]propyl]-5-[methyl(methylsulfonyl)amino]-N¹-[(1R)-1-phenylpropyl]benzene-1,3-dicarboxamide} containing a hydroxyethylamine isostere was determined. Based on this structure, a computational docking study was performed which led to inhibitor 2 a -bound BACE2 models. These were used to optimize the potency and selectivity of inhibitors. A systematic structure–activity relationship study led to the identification of determinants of the inhibitors’ potency and selectivity toward the BACE2 enzyme. Inhibitors 2 d [N³-[(1S,2R)-1-benzyl-2-hydroxy-3-[[(1S,2S)-2-hydroxy-1-(isobutylcarbamoyl)pentyl]amino]propyl]-N¹-methyl-N¹-[(1R)-1-phenylpropyl]benzene-1,3-dicarboxamide; K_i=0.031 nm , selectivity over BACE1: ≈174 000-fold] and 3 l [N¹-((2S,3R)-3-hydroxy-1-phenyl-4-((3-(trifluoromethyl)benzyl)amino)butan-2-yl)-N³,5-dimethyl-N³-((R)-1-phenylethyl)isophthalamide; K_i=1.6 nm , selectivity over BACE1: >500-fold] displayed outstanding potency and selectivity. Inhibitor 3 l is nonpeptide in nature and may pave the way to the development of a new class of potent and selective BACE2 inhibitors with clinical potential.     

Identification of miR-20a-5p as Robust Normalizer for Urine microRNA Studies in Renal Cell Carcinoma and a Profile of Dysregulated microRNAs

Renal cell carcinoma (RCC) is the third most frequent urinary malignancy and one of the most lethal. Current diagnostic and follow-up techniques are harmful and unspecific in low-grade tumors. Novel minimally invasive markers such as urine microRNAs (miRNAs) are under study. However, discrepancies arise among studies in part due to lack of consent regarding normalization. We aimed to identify the best miRNA normalizer for RCC studies performed in urine samples together with a miRNA profile with diagnostic value and another for follow-up. We evaluated the performance of 120 candidate miRNAs in the urine of 16 RCC patients and 16 healthy controls by RT-qPCR followed by a stability analysis with RefFinder. In this screening stage, miR-20a-5p arose as the most stably expressed miRNA in RCC and controls, with a good expression level. Its stability was validated in an independent cohort of 51 RCC patients and 32 controls. Using miR-20a-5p as normalizer, we adjusted and validated a diagnostic model for RCC with three miRNAs (miR-200a-3p, miR-34a-5p and miR-365a-3p) (AUC = 0.65; Confidence Interval 95% [0.51, 0.79], p = 0.043). let-7d-5p and miR-205-5p were also upregulated in patients compared to controls. Comparing RCC samples before surgery and fourteen weeks after, we identified let-7d-5p, miR-152-3p, miR-30c-5p, miR-362-3p and miR-30e-3p as potential follow-up profile for RCC. We identified validated targets of most miRNAs in the renal cell carcinoma pathway. This is the first study that identifies a robust normalizer for urine RCC miRNA studies, miR-20a-5p, which may allow the comparison of future studies among laboratories. Once confirmed in a larger independent cohort, the miRNAs profiles identified may improve the non-invasive diagnosis and follow-up of RCC.     

Differential scanning calorimetry thermal properties and oxidative stability indices of microwave heated extra virgin olive oils

BACKGROUND: The use of differential scanning calorimetry (DSC) for assessing the deterioration effect of microwave heating on vegetable oils, and on olive oils in particular, has been partially explored in literature. The aim of this work was to evaluate the potential of DSC to discriminate among microwaved extra virgin olive oils (EvOo from different olive cultivar and origin), according to changes on thermal properties (upon cooling and heating) and traditional oxidative stability indices (peroxide, p‐anisidine and TOTOX values). RESULTS: An elevated value of lipid oxidation was reached by the most unsaturated EvOo sample (9.5% of linoleic acid) at 6 min of microwave treatment. Free acidity significantly increased (0.42%) only for the oil sample with the highest water content (874 mg kg^?1 oil) at the longest time of treatment. Crystallisation enthalpies significantly decreased and the major exothermic peak shifted towards lower temperature, leading to enlargement of the transition range in all samples due to the formation of weak and mixed crystals among triacylglycerols and lipid degradation products. On the contrary, thermal properties upon heating appeared to similarly vary among samples. CONCLUSIONS: The analysis of DSC thermal properties upon cooling seemed to clearly discriminate among different EvOo samples after microwaving. The relation between changes of thermal properties and oxidation parameters should be further studied using additional oxidative stability indices on a larger set of oil samples, due to the complexity of EvOo composition. Copyright © 2010 Society of Chemical Industry     

Cross-sector collaboration within Dutch flood risk governance: historical analysis of external triggers

ABSTRACT

This article synthesizes the literature on Dutch flood risk governance to analyze how external conditions shaped past and present dynamics of cross-sector collaboration for integrated flood risk management in the Netherlands. It traces the extent to which policy and legal frameworks, socio-economic circumstances, political realities, power relations and conflict situations have influenced attempts at collaboration between flood safety, spatial planning, environmental protection and other sectors. Despite the growing interdependences, existing power relations between the sectors are characterized by the dominance of the water sector. Hence, cross-sector collaboration can develop as long as it does not compromise flood safety.     

Incorporation of calcium salts into xanthan gum matrices: hydration,erosion and drug release characteristics

Xanthan gum (XG), a hydrophilic biopolymer with modified release properties, was used to produce directly compressed matrix tablets containing a model drug, sodium p-aminosalicylate. Three formulations were prepared, each containing a different calcium dihydrate salt: calcium chloride, calcium sulfate or dibasic calcium phosphate. The aim of the investigation was to relate the calcium ion content and solubility of the calcium salt to the in vitro drug release profile of the xanthan matrices. Tablet hydration, erosion and drug release were determined in distilled water using the British Pharmacopoeia (BP) paddle method. The data showed that the overall drug release was the greatest with addition of calcium sulfate, followed by calcium chloride and dibasic calcium phosphate. The chloride salt formulation displayed the greatest percentage erosion due to rapid mass loss during the initial phase, followed by those with sulfate or phosphate salts. As xanthan gel viscosity increased and drug release was also found to be lower, it can be concluded that drug release is influenced by the solubility of the salt present in the formulation, since these parameters determine the viscosity and structure of the gel layer.