The Polyherbal Wattana Formula Displays Anti-Amyloidogenic Properties by Increasing α-Secretase Activities

PLoS One. 2017; 12(1): e0170360. Published online 2017 Jan 20. doi: 10.1371/journal.pone.0170360 PMCID: PMC5249131 Htut Htut Htoo,1 Suveerawan Limsuvan,2 Onusa Thamsermsang,2 Jean-François Hernandez,3 Frédéric Checler,4 Piyarat Govitrapong,1,5,6 Narawut Pakaprot,7 Pravit Akarasereenont,2,8 and Bruno Vincent1,6,9,* Madepalli K. Lakshmana, Editor 1Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom, Thailand 2Center of Applied Thai Traditional Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand 3Institut des Biomolécules Max Mousseron, UMR5247 CNRS/Université de Montpellier/ENSCM, Faculté de Pharmacie, Montpellier, France 4Université Côte d’Azur, INSERM, CNRS, IPMC, Laboratory of excellence DistALZ, Sophia-Antipolis, Valbonne, France 5Center for Neuroscience and Department of Pharmacology, Faculty of Science, Mahidol University, Bangkok, Thailand 6Research Center for Neuroscience, Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom, Thailand 7Department of Physiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand 8Department of Pharmacology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand 9Centre National de la Recherche Scientifique, Paris, France Torrey Pines Institute for Molecular Studies, UNITED STATES Competing Interests: The authors have declared that no competing interests exist. Conceptualization: BV. Data curation: HHH BV. Formal analysis: BV. Funding acquisition: BV PG. Investigation: HHH BV. Project administration: HHH BV. Resources: JFH FC SL OT PA. Supervision: PA BV. Validation: HHH BV. Visualization: HHH BV. Writing – original draft: BV. Writing – review & editing: HHH JFH FC NP PA BV. * E-mail: ht.ca.lodiham@niv.onurb Abstract Alzheimer’s disease is characterized by the deposition of insoluble amyloid-β peptides produced from the β-amyloid precursor protein (βAPP). Because α-secretase cleavage by ADAM10 and ADAM17 takes place in the middle of Aβ, its activation is considered as a promising anti-AD therapeutic track. Here we establish that the polyherbal Wattana formula (WNF) stimulates sAPPα production in cells of neuronal and non-neuronal origins through an increase of both ADAM10 and ADAM17 catalytic activities with no modification of BACE1 activity and expression. This effect is blocked by specific inhibition or genetic depletion of these disintegrins and we show that WNF up-regulates ADAM10 transcription and ADAM17 maturation. In addition, WNF reduces Aβ40 and Aβ42 generation in human cell lines. Altogether, WNF presents all the characteristics of a potent preventive anti-Alzheimer formula. Importantly, this natural recipe, currently prescribed to patients for the treatment of other symptoms without any secondary effect, can be tested immediately for further clinical studies. Go to: Introduction Alzheimer’s disease (AD) is a progressive and yet incurable neurodegenerative disorder affecting the elderly. This syndrome, at its early stage, is characterized by mild memory loss before evolving to a severe decline of cognitive functions and ultimately leading to dementia and death. At the molecular level, proteolysis of the β-amyloid precursor protein (βAPP) by enzymes called “secretase” is a central event since it determines both the production rate and the nature of the amyloid peptide (Aβ) [1], the main component of the extracellular senile plaques formed in the brain of affected individuals. On one hand, the so-called “amyloidogenic” pathway leading to Aβ production is a two-step mechanism along which the β-secretase BACE1 (Beta-site APP-Cleaving Enzyme 1) first generates its N-terminal extremity before Aβ generation is completed by the heterotetrameric γ-secretase complex that liberates the C-terminal end of the peptides. On the other hand, there exists an alternative “non amyloidogenic” route of βAPP processing that is mainly performed by ADAM10 and ADAM17, two members of the ADAM (A Disintegrin And Metalloprotease) family. Importantly, this cleavage of βAPP triggers two beneficial effects regarding AD since it is not only taking place in the middle of the Aβ sequence, thereby preventing its production, but also gives rise to a large sAPPα secreted fragment that displays neuroprotective, neurotrophic, memory-enhancing and neurogenesis-stimulating properties [2–6]. For these reasons, sAPPα is largely considered as a key positive factor in terms of AD. On a therapeutic point of view, besides the promising but yet unsuccessful vaccination approach, numerous studies aimed at inhibiting the amyloidogenic β- and γ-secretases or activating the non-amyloidogenic α-secretases were conducted during the past years [7, 8]. However, acute pharmacological modulation of α-secretases, that are implicated in a wide range of AD-unrelated and important physiological processes, is hardly conceivable as an anti-AD therapy in humans since it would likely trigger some serious side effects, thereby impairing the therapeutic success of such molecules [9]. It has been proposed that an alternative to pharmaceutical therapies could be to regulate these proteases through the regular consumption of natural compounds (nutrients, plant constituents or herbal extracts) that would operate in a mild but chronic manner throughout life (for review see [10]). The Thai Wattana formula (WNF), a polyherbal mixture, has been traditionally used for health prevention and nourishment from age-related problems like loss of appetite, weakness, digestion and gastrointestinal problems. This formula is composed of 15 medicinal plants: Aegle marmelos (L.) Corrêa (Bael fruit), Boesenbergia rotunda (L.) Mansf. (Krachai), Cinnamomum ilicioides A. Chev. (Ka Thon), Cladogynosorientalis Zipp. ex Span. (Chetta Phang Khi), Cryptolepis dubia (Burm.f.) M.R.Almeida (Soften tendons), Cyperusrotundus L. (Nutgrass), Derris scandens (Roxb.) Benth. (Jewel Vine), Putrangiva roxburghii Wall. (Ma Kham Kai), a gum of Ferula assafoetida L. (Asafetida), Ligusticum sinense Oliv. (Kot Hua Bua), Mallotusrepandus (Willd.) Mull.Arg. (Kho Khlan/Fishberry), Piper nigrum L. (White Pepper), Aucklandia lappa DC. (Costus), Terminalia chebula Retz. (Myrobalans) and Tinospora crispa (L.) Hook.f. (Bora Phet) [11]. It is generally prescribed as an appetite stimulant and for health improvement and promotion. Moreover, it has recently been tested for its ability to control the immunomodulatory activity of natural killer and monocyte-derived dendritic cells, osteoarthritis and gastric emptying rate (GER) [12–15]. Finally, it has been shown to convey protection against ultraviolet A-induced melanogenesis through an antioxidant/redox mechanism [11]. Because this formula was also proposed to have some anti-aging properties, we examined its possible beneficial effect in an AD-related context. Here we demonstrate that WNF promotes the non amyloidogenic α-secretase processing of βAPP in neuronal and non-neuronal cells through the activation of ADAM10 and ADAM17 catalytic activities without interfering with β-secretase (BACE1) activity and expression. We also show that WNF regulates these two disintegrins through distinct transcriptional and post-transcriptional mechanisms. Finally, we establish that WNF is able to reduce amyloid peptides production in cells in vitro. Go to: Materials and methods Materials DMEM, Opti-MEM, geneticin, fetal bovine serum (FBS) and lipofectamine 2000 were from Invitrogen (Carlsbad, CA, USA). Penicillin-streptomycin mix was from PAA. Tris and Glycine were from Vivantis (Selangor Darul Ehsan, Malaysia). PDBu, poly-D-lysine, GI254023X and dimethyl sulfoxide were from Sigma (St Louis, MO, USA). Skim milk powder was from HiMedia (Mumbai, India). Ammonium persulphate was from GE Health care (Pisataway, NJ, USA). The chemiluminescence HRP substrate was from Millipore (Bedford, MA, USA). SDS was from Amaresco (Solon, OH, USA). O-Phenanthroline and TAPI-0 were from Calbiochem (San Diego, CA, USA). Preparation and analysis of WNF The sources of all herbal components came from the wild stretching from the Central and the Northeastern parts of Thailand by contact suppliers who collected and sold the crude herbs to the Center of Applied Thai Traditional Medicine where the origin of each herb was recorded. All the production procedures were then supported by the Manufacturing Unit of Herbal Medicines and products, Center of Applied Thai Traditional Medicine (CATTM), Faculty of Medicine Siriraj Hospital (Bangkok) and were operated under Good Manufacturing Practice (GMP) certification. Briefly, individual herbs were first authenticated by experts, including certified pharmacognosists of the Center of applied Thai traditional medicine. All of raw materials were washed with de-ionized water (DI), dried by hot-air oven and then grinded, sieved and packed in laminated vacuum packaging bags. The polyherbal formula powder is then obtained by extraction of equal amount of each herb (weight/weight) with an 80% ethanol solution at a final concentration of 100mg/ml, filtered through cotton wool and subsequently centrifuged at 10,000xg for 10 min. The supernatant was evaporated and lyophilized to obtain freeze-dried powder and stored in amber bottle at 25°C in desiccators. The physical properties, the heavy metal and microbial contamination of formula were assessed before any experiment is performed. In addition, the chemical assessments of the formula were verified using Thin Layer Chromatography (TLC) and Ultra Performance Liquid Chromatography (UPLC) as previously described [11]. WNF was freshly prepared before experimental use on cell lines as a 10mg/ml stock solution in 50% DMSO. Cell lines, transfections and in vitro treatments Human HEK293 cells and mouse embryonic fibroblasts (MEFs) were cultured at 37°C, 5% CO2 in DMEM supplemented with 10% FBS, penicillin (100U/ml) and streptomycin (50mg/ml). HEK293 cells stably overexpressing human βAPPwt751, human ADAM10, mouse ADAM17 or 1D4-tagged human BACE1 were obtained and maintained as previously described [16–18]. Mouse neuroblastoma N2a cells stably overexpressing human wild-type neuronal βAPP695 [19] were cultured at 37°C, 5% CO2 in 50% DMEM, 50% Opti-MEM, supplemented with 5% FBS, antibiotics and geneticin (0.25g/l). MEFs derived from ADAM10-/- or ADAM17-/- mice (as well as wild-type controls) were previously described [20, 21]. Human SH-SY5Y neuroblastoma cells were grown at 37°C, 5% CO2 in 50%DMEM/50%F12 containing 10% FBS, glutaMAX (2mM), non-essential amino-acids (0.05%) and antibiotics. Transient overexpression of human wild-type βAPP751 in MEFs, HEK293 and SH-SY5Y cells was carried out with lipofectamine 2000. WNF was prepared as a 10mg/ml solution by adding 1 ml of 50% DMSO/50% H2O to 10mg of desiccated pellet and used for the 100μg/ml treatments. Serial dilutions were performed for lower concentrations. For all conditions (including non-treated controls), DMSO was adjusted to 0.5%. sAPPα secretion and western blot analysis Secretion and detection of sAPPα in HEK293, N2a, SH-SY5Y and MEFs with the human-specific monoclonal DE2B4 antibody (dilution 1/500, Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA) has been previously described [22]. Cells were collected with phosphate-buffered saline (PBS)-EDTA and resuspended in 80 to 150 μl of lysis buffer (10mM Tris/HCl, pH 7.5, 150mM NaCl, 0.5% triton X-100, 0.5% deoxycholate, 5mM EDTA). Protein concentrations were determined by the Bradford method [23] and proteins were separated by SDS-PAGE on 8% (βAPP, calnexin and ADAM17) or 10% Tris/glycine (ADAM10, BACE1, Golgi 58K protein and β-actin), or 16.5% Tris/tricine gels (C83). Proteins were transferred onto nitrocellulose membranes (45 to 120min according to protein size, 100V), blocked for 1h in 5% non-fat milk and incubated overnight at 4°C with primary antibodies directed against βAPP/C83 (dilution 1/4000, polyclonal A8717, Sigma), ADAM10 (dilution 1/500, polyclonal AB19026, Millipore, Bedford, MA, USA), ADAM17 (dilution 1/1000, polyclonal AB19027, Millipore), BACE1 (dilution 1/1000, monoclonal EPR3956, Abcam, Cambridge, UK), calnexin (dilution 1/1000, monoclonal 610523, BD Biosciences, Singapore), Golgi 58K protein (dilution 1/2000, monoclonal G2404, Sigma) or β-actin (dilution 1/5000, monoclonal 13E5, Cell Signaling, Beverly, MA, USA). Bound antibodies were detected using goat anti-mouse (dilution 1/3000, polyclonal 7076, Cell Signaling) or goat anti-rabbit peroxidase-conjugated antibody (dilution 1/3000, polyclonal 7074, Cell Signaling). Immunological complexes were revealed using ECL methods (Immobilon Western Chemiluminescence HRP substrate) and detected using an automatic medical X-ray processor (Kodak, Rochester, NY, USA). Bands densitometries were measured with the ImageJ software and normalized using β-actin as internal standard. α-secretase fluorimetric assay on intact cells Wild-type, ADAM10-/- or ADAM17-/- MEFs as well as HEK293 cells overexpressing ADAM10 or ADAM17 were cultured in 35mm-dishes coated with polylysine (10μg/ml) until cells reached 80% confluence. Cells were treated in duplicate without (control) or with WNF (100μg/ml) for 16 hours at 37°C in 1ml of DMEM containing 1% FBS. After this treatment period, the PKC activator phorbol 12, 13-dibutyrate (PDBu) (1μM, wild-type/ADAM17-/- MEFs and ADAM17-overexpressing HEK293 cells) was added for two hours. Duplicates were then incubated for 30 min at 37°C in the absence or in the presence of o-phenanthroline (general metalloprotease inhibitor, 100μM), GI254023X (ADAM10-specific inhibitor, 10μM), or TAPI-O (ADAM17-specific inhibitor, 10μM) in 1.5ml of PBS. Then, the α-secretase-specific JMV2770 substrate (10μM) [24] was directly added into the media and cells were maintained at 37°C. At each time point, 100μl of media were removed and the α-secretase-specific activity corresponding to the o-phenanthroline-, GI254023X- or TAPI-O-sensitive fluorescence was recorded in black 96-well plates at 320nm and 420nm excitation and emission wavelengths respectively. β-secretase fluorimetric assay on cell homogenates HEK293 cells stably overexpressing 1D4-BACE1 were cultured in 35mm-dishes until they reach 80% confluence, treated without (control) or with WNF (100μg/ml) for 16 hours at 37°C in DMEM containing 1% FBS and assayed for their β-secretase activity as previously described [16]. Briefly, cells were collected, lysed with Tris 10mM pH 7.5, homogenized and kept on ice. Samples were assayed for their protein contents with the Bradford method and all adjusted to a 3μg/μl concentration. Thirty μg of each samples (10μl) diluted in 10mM sodium acetate buffer pH 4.5 were incubated for 30 min at 37°C in black 96-well plates (in a final volume of 100μl) in the absence (triplicate) or in the presence (triplicate) of the β-secretase specific inhibitor JMV1197. Then, the β-secretase-specific JMV2236 substrate (10μM) was added to all samples and the β-secretase-specific activity corresponds to the JMV1197-sensitive fluorescence recorded at each time point at 320nm and 420nm excitation and emission wavelengths respectively. Real-time quantitative polymerase chain reaction (q-PCR) Following treatments (36 hours), total RNA was extracted and purified with the PureLink RNA mini kit (Ambion, Life Technologies, Austin, TX, USA). Real-time PCR was performed with 100ng of total RNA using the QuantiFast SYBR Green RT-PCR kit (Qiagen, Singapore) detector system (Eppendorf Mastercycler ep RealPlex) and the SYBR Green detection protocol. The 2x QuantiFast SYBR Green RT-PCR master mix, QuantiFast RT mix, QuantiTectPrimer Assay and template RNA were mixed and the reaction volume was adjusted to 25μl using RNase-free water. The specific primers were designed and purchased from Qiagen. Each primer is a 10x QuantiTect Primer Assay containing a mix of forward and reverse primers for specific targets: Hs_ADAM10_1_SG (QT00032641, human ADAM10), Hs_ADAM17_1_SG (QT00055580, human ADAM17) and Hs_GAPDH_1_SG (QT00079247, human GAPDH). Sucrose gradient sub cellular fractionation Wild-type HEK293 cells cultured in 100mm-dishes were incubated for 16 hours without (control) or with WNF (100μg/ml) in DMEM/1% FBS. Cells were then homogenized with a dounce homogenizer in 0.25M sucrose prepared in 10mM Tris-HCl (pH 7.4) and containing 1mM Mg(AcO)2. Equal amounts of protein were loaded at the top of a step gradient, centrifuged and fractions (1ml) were collected from top to bottom of each gradient. Proteins in fractions were precipitated overnight at 4°C with methanol (4 volumes) and calnexin, Golgi 58K protein, ADAM10 and ADAM17 immunoreactivities were analyzed by western blot as described above. Measurement of human Aβ production HEK293 and SH-SY5Y neuroblastoma cells were transiently transfected with the human wild-type βAPP751 cDNA for 36 hours and incubated without (control) or with various WNF concentrations during the last 16 hours (in 1ml of DMEM/FBS1%). Aβ40 and Aβ42 levels were then detected in the secretion media (50μl) using sandwich ELISA kits detecting human Aβ40 (khb3482) and human Aβ42 (khb3442) respectively (Invitrogen) following manufacturer’s recommendations. Aβ levels (pg/ml) were obtained by reading absorbance at 450nm with a spectrophotometer and values were then normalized with βAPP and β-actin. Statistical analysis Statistical analyses were performed with the Prism software (GraphPad, San Diego, USA) using the unpaired t test for pair wise comparisons. All results were expressed as means ± SEM and the p values equal to or less than 0.05 were considered significant. Go to: Results We first evaluated the effect of WNF on the non-amyloidogenic α-secretase processing of βAPP by cultured human HEK293 cells overexpressing βAPP751 and showed that concentrations from 1 up to 100μg/ml dose-dependently increase the secretion of sAPPα as well as the production of the α-secretase-derived C-terminal counterpart (C83 fragment) without modifying βAPP immunoreactivity (Fig 1A), thereby indicating that WNF most likely up-regulates the α-secretase processing of βAPP rather than βAPP expression. Considering the well-established roles of ADAM10 and ADAM17 in the constitutive and PKC-regulated α-secretase processing of βAPP respectively [25], we tested the effects of the ADAM10-specific inhibitor GI254023X as well as the ADAM17-specific inhibitor TAPI-O on the WNF-induced sAPPα production. The results showed that GI254023X and TAPI-O respectively prevent the constitutive (Fig 1B) and PDBu-stimulated (Fig 1C) WNF-dependent sAPPα secretion in βAPP-overexpressing HEK293 cells. Because these experiments were conducted in HEK293 cells artificially overexpressing high amounts of the βAPP protein, we then wanted to determine whether WNF was able to generate similar effects in the same cell line producing endogenous levels of βAPP. Indeed, WNF dose-dependently and significantly promotes sAPPα release without interfering with βAPP protein levels in wild-type HEK293 cells (Fig 1D). Fig 1 Fig 1 WNF stimulates sAPPα production in human HEK293 cells. We then validated these data in cells of neuronal origin and carried out similar experiments with cultured mouse N2a neuroblastoma cells stably overexpressing the neuron-specific βAPP695 isoform. As observed in HEK293 cells, WNF stimulates the secretion of sAPPα in a dose-dependent manner without altering βAPP levels (Fig 2A). We then wanted to ascertain that the formula conveys similar effects on the endogenous α-secretase processing of βAPP and established that WNF significantly increases endogenous sAPPα secretion in a dose-dependent manner in human SH-SY5Y neuroblastoma cells (Fig 2B). Fig 2 Fig 2 WNF promotes sAPPα production in mouse and human neuroblastoma cell lines. We then used a specific and reliable α-secretase-specific fluorimetric assay [24] to investigate the effect of WNF (100μg/ml) on ADAM10 and ADAM17 catalytic activities. We observed that WNF triggers a significant increase of both the constitutive (ADAM10-expressing HEK293 cells) (Fig 3A) and PKC-regulated (PDBu-treated ADAM17 expressing cells) (Fig 3B) o-phenanthroline-sensitive JMV2770-hydrolyzing activities. Similar experiments performed using GI254023X (ADAM10-specific) and TAPI-O (ADAM17-specific) inhibitors indicated that WNF significantly triggers both GI254023X- (Fig 3C) and TAPI-O-sensitive (Fig 3D) JMV2770 degradation and confirmed that WNF indeed targets ADAM10 and ADAM17. We also evaluated the impact of WNF treatment on the amyloidogenic BACE1 catalytic activity by means of a specific fluorimetric assay [16]. As shown in Fig 3E, WNF applied at a concentration of 100μg/ml did not affect the JMV1197-sensitive JMV2236 degradation in BACE1-overexpressing HEK2983 cells. Moreover, WNF treatments did not modify endogenous BACE1 immunoreactivity in wild-type HEK293 cells (Fig 3F). Fig 3 Fig 3 WNF increases α-secretase catalytic activities in HEK293 cells without interfering with BACE1 activity and expression. We next examined the ability of WNF to stimulate sAPPα production in the previously well-characterized MEFs derived from wild-type, ADAM10-/- and ADAM17-/- animals [20–22]. Because the anti-sAPPα antibody DE2B4 is human-specific, we transiently transfected human βAPP751. As shown in Fig 4A and 4B, the three cell lines efficiently overexpressed human βAPP and secreted detectable amounts of human sAPPα at 36 hours post-transfection. Importantly, WNF (100μg/ml) could promote the constitutive (Fig 4A, upper left panel) and PKC-regulated (Fig 4B, upper left panel) release of sAPPα in the wild-type MEFs, but not in ADAM10 and ADAM17 knockout cells respectively (Fig 4A and 4B, upper right panels). Moreover, the positive effects of WNF toward both the constitutive (Fig 4C, left panel) and PDBu-induced (Fig 4D, left panel) JMV2770-hydrolyzing activities observed in wild-type MEFs were fully prevented by ADAM10 (Fig 4C, right panel) and ADAM17 depletions (Fig 4D, right panel) respectively. Altogether, these data undoubtedly established ADAM10 and ADAM17 as genuine mediators of WNF-dependent α-secretase activation. Fig 4 Fig 4 ADAM10 and ADAM17 knockouts respectively abolish the constitutive and PDBu-stimulated WNF-induced sAPPα secretions and JMV2770 hydrolysis in mouse embryonic fibroblasts. To determine whether WNF up-regulates ADAM10 and ADAM17 expressions, we first measured the impact of WNF treatment on ADAM10 and ADAM17 protein levels in HEK293 cells by western blot. Surprisingly, WNF (100μg/ml) significantly increased ADAM10 (Fig 5A, left panels) but not ADAM17 endogenous immunoreactivities (Fig 5A, right panels). We next performed quantitative real-time PCR experiments and established that WNF treatment leads to a significant augmentation of ADAM10 but not ADAM17 mRNA levels (Fig 5B). Because WNF apparently triggers ADAM17 maturation as illustrated by an increase of active ADAM17 (lower band, Fig 5A upper right panel), velocity sedimentation of ADAM10 and ADAM17 in sucrose step gradients was performed. Partial characterization of the fractions, using anti-58K Golgi protein and anti-calnexin antibodies as markers of the Golgi apparatus and the endoplasmic reticulum (ER), indicated that these organelles reside in fractions 1–3 and fractions 11–12 respectively (Fig 5C panels e and f). The results indicated that whereas the distribution profiles of endogenous ADAM10 was similar in control and WNF-treated cells (Fig 5C panels a and b), the maturation of endogenous ADAM17 was strongly enhanced by WNF, thereby increasing ADAM17 maturation in the Golgi/trans-Golgi network (TGN) (compare mature ADAM17 immunoreactivity in fractions 1–3 for panels c and d in Fig 5C) as well as the level of active ADAM17 in the ER/plasma membrane fractions (Fig 5C, lanes 11 and 12 in panels c and d). Fig 5 Fig 5 WNF up-regulates ADAM10 and ADAM17 via two distinct mechanisms in human HEK293 cells. Finally, because α-secretase cleaves βAPP in the middle of the Aβ sequence, we hypothesized that WNF treatment could affect amyloid peptides generation in non-neuronal and neuronal human cells transiently overexpressing human βAPP. Firstly, thirty-six hours post-transfection, both HEK293 and SH-SY5Y cells express high βAPP levels when compared to pcDNA3-transfected cells (Fig 6A and 6B upper panels). Secondly and remarkably, WNF applied at a dose of 100μg/ml significantly reduced the secretion of Aβ40 as well as the toxic and aggregate-prone Aβ42 peptide in HEK293 cells (Fig 6A) as well as in the SH-SY5Y neuroblastoma cell line (Fig 6B). Fig 6 Fig 6 WNF decreases Aβ peptides generation in HEK293 and SH-SY5Y human cells. Go to: Discussion Together with Aβ-targeting vaccination, the pharmacological inhibition of the two β- and γ-secretases Aβ-forming enzymes stood during the past decades as the principal and most relevant therapeutic tracks aimed at preventing, slowing down or curing Alzheimer’s disease. However, because BACE1 and γ-secretase cleave, in addition to βAPP, a constantly growing number of other substrates with important physiological functions, this strategy may engender severe deleterious side effects. Considering this matter of fact, a more recently developed alternative consists in the activation of the α-secretases ADAM10 and ADAM17. The principal advantage of such an approach as an anti-AD therapy resides in the fact that stimulation of this cleavage is not only expected to preclude Aβ production but also to support neurotrophism, neuroprotection and neurogenesis through an increased secretion of the βAPP-derived sAPPα metabolite. Unfortunately, because ADAM10 and/or ADAM17 have more than 80 other substrates, the cleavages of which yielding to pathological situations such as cancer and chronic inflammation [9], one should stay very cautious regarding the use of acute pharmacological activation of α-secretases and to rather envision stimulating these proteases via the mild, safe and regular consumption of natural compounds that would reduce the amyloid load on a long term basis and could thereby represent a valuable therapeutic alternative for AD treatment [26]. Falling within such an approach, the present study proposed to investigate the effect of the polyherbal Wattana formula on the non-amyloidogenic processing of βAPP in vitro in various cell lines by means of complementary techniques aimed at measuring sAPPα secretion, Aβ production as well as α-secretase catalytic activities, expression and subcellular distribution. We first established that WNF stimulates the constitutive and the PKC-regulated α-secretase activities in a dose-dependent manner. Importantly, WNF not only stimulates sAPPα secretion in βAPP-overexpressing non-neuronal HEK293 and neuroblastoma N2a cells, thereby indicating its ubiquitous action, but also behaves as a potent enhancer of endogenous sAPPα production in the human SH-SY5Y neuroblastoma cell line. In addition, using pharmacological inhibition and genetic depletion approaches, we formally identified ADAM10 and ADAM17 as the targeted proteases, thereby establishing WNF as a potent α-secretase enhancer and a possible anti-AD agent. It is important to underline here that, as far as the amyloid cascade hypothesis is considered to be at the center of gravity of the pathology, α-secretase natural activators in general and WNF in particular are expected to bring preventive rather than curative beneficial effects since they theoretically impair all the following events such as Aβ oligomerization and fibrillogenesis as well as the subsequent cognitive impairments associated with the disease. Our observations that WNF does not interfere with the amyloidogenic β-secretase activity but significantly lowers both Aβ40 and Aβ42 production underline the fact that any therapeutic strategy leading to α-secretase activation with no modification of β-secretase activity is expected to be sufficient to impair Aβ generation and is in good agreement with the observation that the sole ADAM10 overexpression reduces both Aβ40 and Aβ42 levels in vivo in the brain of a transgenic mouse model of AD [27]. On a mechanistic point of view, it is of utmost interest to underline that ADAM10 and ADAM17, although presenting a similar general structure, are up-regulated by WNF via two distinct mechanisms. Thus, WNF induces an elevation of ADAM10 immunoreactivity and mRNA levels thereby demonstrating an effect at the transcriptional level. However, the same treatment has no impact on ADAM17 transcription but rather promotes its maturation/activation as shown by the marked increase of active ADAM17 and the concomitant decrease of the pro-enzyme (Fig 4C, bottom panels). Because WNF is a mixture of 15 medicinal plants, the most probable explanation would be that ADAM10 and ADAM17 are targeted by distinct WNF components, the identification of which remaining to be established. In this respect, some of the molecules identified as part of the WNF by ultra-performance liquid chromatography [14] were indeed recently shown to be beneficial regarding AD pathology. Firstly, the active alkaloid and acetylcholinesterase inhibitor piperine can protect against neurodegeneration and cognitive impairment in a rat model of AD [28]. Secondly, the antioxidant and anti-inflammatory polyphenol gallic acid is able to decrease Aβ toxicity [29], to reduce amyloid fibril formation [30] and to attenuate neuronal damage by preventing Aβ oligomerization [31]. Thirdly, the antioxidant and NFκB inhibitor p-coumaric acid as well as the phenolic compound caffeic acid protect against Aβ25–35-induced neurotoxicity respectively in vitro in PC12 cells [32] and in vivo in rats [33]. Fourthly, ferulic acid, a phenol that is closely related to curcumin and has antioxidant properties, induces a resistance to Aβ42 toxicity in adult mice [34] and reduces amyloid deposition in a mouse model of AD [35]. However, contrary to our findings, all these described effects most likely occur at a late post-Aβ production step rather than at earlier βAPP processing stages. Nevertheless, two recent publications have evidenced that some of these compounds can indeed target βAPP-cleaving secretases. Hence, ferulic acid can reverse the behavioral deficits of the PSAPP transgenic mouse model of AD through a slight reduction of the β-secretase BACE1 stability and activity [36] that we could not detect in the whole extract. Moreover, octyl gallate, the ester of 1-octanol and gallic acid, has been shown to inhibit Aβ generation and to increase sAPPα secretion in vitro and in vivo via an increase of estrogen receptor-mediated ADAM10 activity [37] and could therefore support the WNF-dependent increase of ADAM10 activity/expression observed in the present study. It is also worth mentioning that besides its herein described α-secretase-stimulating property, WNF may also provide an additional and complementary beneficial anti-AD effect since five of its components (Aucklandialappa DC. (Costus); Terminalia chebula Retz. (Myrobalans); Piper nigrum L. (White Pepper); Cyperusrotundus L. (Nutgrass) and Aegle marmelos (L.) Corrêa (Bael fruit)) display acetylcholinesterase inhibitory activity [38–40]. Supporting the promising anti-AD therapeutic use of medicinal plants, several other plant extracts have been recently reported to convey beneficial effects in vivo in animal models of AD ([41] for review). Firstly, extracts prepared from Centella asiatica (L.) Urb. (Umbelliferae) were shown to ameliorate cognitive performances and to decrease Aβ levels, oxidative stress and senile plaques formation [42, 43]. Secondly, extract from Bacopa monnieri (L.) Wettst. (Brahmi) displays neuroprotective effects, reduces Aβ production and can improve cognitive functions [44, 45]. Finally, extracts from Withania somnifera (L.) Dunal diminish behavioral deficits, Aβ production and plaque pathology [46]. Following the present demonstration that WNF is able to increase sAPPα secretion and to reduce Aβ production in vitro, evaluating whether individuals who regularly consume WNF are less prone to developing AD will be of particular interest. Go to: Conclusion This study clearly establishes WNF as a potent in vitro activator of the non-amyloidogenic processing of βAPP. Because this is accompanied by a reduction of amyloid peptides production, it is our assumption that WNF may be used as a mild natural anti-AD preventive treatment. The identification of the WNF-containing active molecule(s) as well as the demonstration that WNF can slow down or reverse the pathology in transgenic mouse models of AD should deserve a particular attention in a near future. However, it is important to underline here that, regardless of this considerations, the use of this polyherbal formula is already possible for further clinical studies in humans since it is currently prescribed to patients for the treatments of AD-unrelated symptoms and does not display any secondary effects. Go to: Acknowledgments We thank Amgen (Seattle, WA, USA) and P. Saftig (University of Kiel, Germany) for providing us with MEFs and H. Xu (Xiamen University, China and Sanford-Burnham Medical Research Institute, La Jolla, CA, USA) for the kind donation of N2aAPP695 cells. Go to: Funding Statement This work was supported by the National Science and Technology Development Agency (NSTDA) grant number P-10-11285 (PG, BV) and grant number P-11-00783 (BV); http://www.nstda.or.th/eng/index.php; the Thailand Research Fund (TRF) grant number IRG 5780009 (BV); http://www.trf.or.th/eng/index.php; and Mahidol University budget grant 2014 (BV); https://www.mahidol.ac.th/en/. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Go to: Data Availability All relevant data are within the paper. Go to: References 1. Zhang H, Ma Q, Zhang YW, Xu H. Proteolytic processing of Alzheimer’s β-amyloid precursor protein. J Neurochem. 2012;120(Suppl.1):9–21. [PMC free article] [PubMed] 2. Mattson MP, Cheng B, Culwell AR, Esch FS, Lieberburg I, Rydel RE. Evidence for excitoprotective and intraneuronal calcium-regulating roles for secreted forms of the β-amyloid precursor protein. Neuron 1993;10:243–254. [PubMed] 3. Furukawa K, Sopher BL, Rydel RE, Begley JG, Pham DG, Martin GM, et al. Increased activity-regulating and neuroprotective efficacy of α-secretase-derived secreted amyloid precursor protein conferred by a C-terminal heparin-binding domain. J Neurochem. 1996;67(5):1882–1896. [PubMed] 4. Claasen AM, Guévremont D, Mason-Parker SE, Bourne K, Tate WP, Abraham WC, et al. 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Familial Alzheimer’s disease-linked presenilin 1 variants elevate Aβ1-42/1-40 ratio in vitro and in vivo. Neuron 1996;17(5):1005–13. [PubMed] 20. Black RA, Rauch CT, Kozlosky CJ, Peschon JJ, Slack JL, Wolfson MF, et al. A metalloproteinase disintegrin that releases tumour-necrosis factor-α from cells. Nature 1997;385(6618):729–33. doi: 10.1038/385729a0 [PubMed] 21. Hartmann D, de Strooper B, Serneels L, Craessaerts K, Herreman A, Annaert W, et al. The disintegrin/metalloprotease ADAM10 is essential for notch signaling but not for α-secretase activity in fibroblasts. Hum Mol Genet. 2002;11(21):2615–24. [PubMed] 22. Shukla M, Htoo HH, Wintachai P, Hernandez JF, Dubois C, Postina R, et al. Melatonin stimulates the nonamyloidogenic processing of βAPP through the positive transcriptional activation of ADAM10 and ADAM17. J Pineal Res. 2015;58(2):151–65. doi: 10.1111/jpi.12200 [PubMed] 23. Bradford MM. A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 1976;72:248–54. [PubMed] 24. Alfa Cissé M, Gandreuil C, Hernandez JF, Martinez J, Checler F, Vincent B. Design and characterization of a novel cellular prion-derived quenched fluorimetric substrate of α-secretase. Biochem Biophys Res Commun. 2006;347(1):254–60. doi: 10.1016/j.bbrc.2006.06.065 [PubMed] 25. Vingtdeux V, Marambaud P. Identification and biology of α-secretase. J Neurochem. 2012;120(Suppl.1):34–45. [PubMed] 26. Essa MM, Vijayan RK, Castellano-Gonzalez G, Memon MA, Braidy N, Guillemin GJ. Neuroprotective effect of natural products against Alzheimer’s disease. Neurochem Res. 2012;37(9):1829–42. doi: 10.1007/s11064-012-0799-9 [PubMed] 27. Postina R, Schroeder A, Dewachter I, Bohl J, Schmitt U, Kojro E, et al. A disintegrin-metalloproteinase prevents amyloid plaque formation and hippocampal defects in an Alzheimer disease mouse model. J Clin Invest. 2004;113(10):1456–64. doi: 10.1172/JCI20864 [PMC free article] [PubMed] 28. Chonpathompikunlert P, Wattanathorn J, Muchimapura S. Piperine, the main alkaloid of Thai black pepper, protects against neurodegeneration and cognitive impairment in animal model of cognitive deficit like condition of Alzheimer’s disease. Food Chem Toxicol. 2010;48(3):798–802. doi: 10.1016/j.fct.2009.12.009 [PubMed] 29. Kim MJ, Seong AR, Yoo JY, Jin CH, Lee YH, Kim YJ, et al. Gallic acid, a histone acetyltransferase inhibitor, suppresses β-amyloid neurotoxicity by inhibiting microglial-mediated neuroinflammation. Mol Nutr Food Res. 2011;55(12):1798–1808. doi: 10.1002/mnfr.201100262 [PubMed] 30. Liu Y, Pukala TL, Musgrave IF, Williams DM, Dehle FC, Carver JA. Gallic acid is the major component of grape seed extract that inhibits amyloid fibril formation. Bioorg Med Chem Lett. 2013;23(23):6336–40. doi: 10.1016/j.bmcl.2013.09.071 [PubMed] 31. Hajipour S, Sarkaki A, Farbood Y, Eidi A, Mortazavi P, Valizadeh Z. Effect of gallic acid on dementia type of Alzheimer’s disease in rats: electrophysiological and histological studies. Basic Clin Neurosci. 2016;7(2):97–106. doi: 10.15412/J.BCN.03070203 [PMC free article] [PubMed] 32. Yoon JH, Youn K, Ho CT, Karwe MV, Jeong WS, Jun M. p-coumaric acid and ursolic acid from Corni fructus attenuated β-amyloid25-35-induced toxicity through regulation of the NFκB signaling pathway in PC12 cells. J Agric Food Chem. 2014;62(21):4911–16. doi: 10.1021/jf501314g [PubMed] 33. Kim JH, Wang Q, Choi JM, Lee S, Cho EJ. Protective role of caffeic acid in an Aβ25-35-induced Alzheimer’s disease model. Nutr Res Pract. 2015;9(5):480–8. doi: 10.4162/nrp.2015.9.5.480 [PMC free article] [PubMed] 34. Yan JJ, Cho JY, Kim HS, Kim KL, Jung JS, Huh SO, et al. Protection against β-amyloid peptide toxicity in vivo with long-term administration of ferulic acid. Brit J Pharmacol. 2001;133(1):89–96. [PMC free article] [PubMed] 35. Yan JJ, Jung JS, Kim TK, Hasan MA, Hong CW, Nam JS, et al. Protective effects of ferulic acid in amyloid precursor protein plus presenilin-1 transgenic mouse model of Alzheimer’s disease. Biol Pharm Bull. 2013;36(1):140–3. [PubMed] 36. Mori T, Koyama N, Guillot-Sestier MV, Tan J, Town T. Ferulic acid is a nutriceutical β-secretase modulator that improves behavioral impairment and Alzheimer-like pathology in transgenic mice. PLoS ONE 2013;8(2):e55774 doi: 10.1371/journal.pone.0055774 [PMC free article] [PubMed] 37. Zhang SQ, Sawmiller D, Li S, Rezai-Zadeh K, Hou H, Zhou S, et al. Octyl gallate markedly promotes anti-amyloidogenic processing of APP through estrogen receptor-mediated ADAM10 activation. PLoS ONE 2013;8(8):e71913 doi: 10.1371/journal.pone.0071913 [PMC free article] [PubMed] 38. Ingkaninan K, Temkitthawon P, Chuenchom K, Yuyaem T, Thongnoi W. Screening for acetylcholinesterase inhibitory activity in plants in Thai traditional rejuvenating and neurotonic remedies. J Ethanopharmacol. 2003;89(2–3):261–4. [PubMed] 39. Asaduzzaman M, Uddin MJ, Kader MA, Alam AHMK, Rahman AA, Rashid M, et al. In vitro acetylcholinesterase inhibitory activity and the antioxidant properties of Aegle marmelos leaf extract: implications for the treatment of Alzheimer’s disease. Psychogeriatrics 2014;14(1):1–10. doi: 10.1111/psyg.12031 [PubMed] 40. Mathew M, Subramanian S. In vitro screening for anti-cholinesterase and antioxidant activity of methanolic extracts of Ayurvedic medicinal plants used for cognitive disorders. PLoS ONE 2014;9(1):e86804 doi: 10.1371/journal.pone.0086804 [PMC free article] [PubMed] 41. Rao RV, Descamps O, John V, Bredesen DE. Ayurvedic medicinal plants for Alzheimer’s disease. Alzheimer’s Res Ther. 2012;4(3):22. [PMC free article] [PubMed] 42. Veerendra Kumar MH, Gupta YK. Effect of Centella asiatica on cognition and oxidative stress in an intracerebroventricular streptozotocin model of Alzheimer’s disease in rats. Clin Exp Pharmacol Physiol. 2003;30(5–6):336–42. [PubMed] 43. Dhanasekaran M, Holcomb LA, Hitt AR, Tharakan B, Porter JW, Young KA, et al. Centella asiatica extract selectively decreases amyloid β levels in hippocampus of Alzheimer’s disease animal model. Phytother Res. 2009;23(1):14–19. doi: 10.1002/ptr.2405 [PubMed] 44. Holcomb LA, Dhanasekaran M, Hitt AR, Young KA, Riggs M, Manyam BV. Bacopa monniera extract reduces amyloid levels in PSAPP mice. J Alzheimers Dis. 2006;9(3):243–51. [PubMed] 45. Uabundit N, Wattanathorn J, Mucimapura S, Ingkaninan K. Cognitive enhancement and neuroprotective effects of Bacopa monnieri in Alzheimer’s disease model. J Ethnopharmacol. 2010;127:26–31. doi: 10.1016/j.jep.2009.09.056 [PubMed] 46. Sehgal N, Gupta A, Valli RK, Joshi SD, Mills JT, Hamel E, et al. Withania somnifera reverses Alzheimer’s disease pathology by enhancing low-density lipoprotein receptor-related protein in liver. Proc Natl Acad Sci U.S.A 2012;109(9):3510–5. doi: 10.1073/pnas.1112209109 [PMC free article] [PubMed] Articles from PLoS ONE are provided here courtesy of Public Library of Science

Is North American Quinoa Finally Here?

http://www.nutritionaloutlook.com/food-beverage/north-american-quinoa-finally-here

Re: Formulation of Olive Oil, Olive Fruit, and Fig Improves Global Health and Well-being in Patients with Rheumatoid Arthritis; Effects on Remission Indicators Are Positive but Not Significant

PDF (Download) Olive (Olea europaea, Oleaceae) Oil Fig (Ficus carica, Moraceae) Rheumatoid Arthritis Date: 08-15-2017 HC# 021734-574 Bahadori S, Salamzadeh J, Kamalinejad M, Shams Ardekani MR, Keshavarz M, Ahmadzadeh A. Study of the effect of an oral formulation of fig and olive on rheumatoid arthritis (RA) remission indicators: a randomized clinical trial. Iran J Pharm Res. 2016;15(3):537-545. Rheumatoid arthritis (RA) is characterized by synovitis and joint bone damage, which cause swelling, pain, morning stiffness, fatigue, and disability, and currently no prevention or cure exists. Treatment to limit permanent joint damage includes the use of disease-modifying antirheumatic drugs (DMARDs) and biologic agents; however, these drugs are not always effective and can cause adverse effects. Olive (Olea europaea, Oleaceae) and fig (Ficus carica, Moraceae) have been found to have beneficial anti-inflammatory, immunomodulatory, antimicrobial, anticancer, analgesic, and antioxidant properties. The goal of this randomized, controlled, parallel-group, clinical trial was to examine the effects of a formulation of olive oil, olive fruit, and fig fruit on RA remission. Conducted from September 2014 to August 2015, the study included patients aged 18 years and older diagnosed with RA and attending the inpatient and outpatient rheumatology departments of Loghman-e Hakim University Hospital at Shahid Beheshti University of Medical Sciences in Tehran, Iran. The patients were randomly assigned to take routine DMARDs in the control group (n=36) or to take routine DMARDs plus a formulation of fig and olive in the intervention group (n=36). The 16-week study included a baseline visit and follow-up visits every 4 weeks, for a total of 5 study visits. The herbal formulation, a semisolid mixture, included olive oil, olive fruit, and fig fruit in a ratio of 2:5:1. Olives and olive oil from trees cultivated at Rudbar City in the Gilan province in north Iran were used for the study. The purchased, dried fig fruits were from trees in Estahban in Iran's Fars province. Patients were instructed to consume 15 grams (approximately 1 tablespoon) of the mixture 3 times daily with meals and to continue their usual diet. The primary outcome was the Disease Activity Score with 28-joint counts based on erythrocyte sedimentation rate (DAS28-ESR) recommended by the American College of Rheumatology. Four components of the score―tender joint count, swollen joint count, ESR, and patient global assessment of disease activity (PtGA)―were measured at each study visit. Also assessed were demographic characteristics, medical and medication history, and any adverse effects. At baseline, all patients had a DAS28-ESR greater than 2.6. Of the 72 patients who were assigned to a study group, 29 in the intervention group and 27 in the control group completed the study. In the intervention group, 7 were lost to follow-up for the following reasons: 3 discontinued treatment, 1 had severe hiccups as an adverse drug reaction, 2 were transient residents, and 1 complained of an unpleasant taste. From the control group, 6 discontinued the intervention, 2 traveled from the area, and 1 was a transient resident. Baseline demographic characteristics, as well as medication and medical histories, were similar between the 2 groups. The authors report a compliance rate of 93.88% ± 7.06% in the intervention group. Results reveal no significant between-group differences in the indicators of RA remission, although positive trends were seen in the intervention group. A greater reduction in PtGA in the intervention group compared with the control group beginning at week 12 indicated a nonlinear significant relationship between PtGA and time (P=0.03). This finding supports "the assumption that a longer course of supplement usage could lead to greater remission in RA, which can be reflected in the values of the disease activity measures," write the authors. No significant between-group differences were observed in the number of patients needing intra-articular glucocorticoids or nonsteroidal anti-inflammatory drugs or in total cumulative dose of steroids during the study. The only adverse event reported was the one patient who withdrew from the intervention group because of severe unclassified hiccups. The authors conclude that although changes in the DAS28-ESR were not significantly different between the 2 study groups, the positive trends in the intervention group and the significant improvements in PtGA scores in the intervention group compared with the control group "convince us to suggest further investigations on the supplementary olive and fig products, with … longer follow up periods." ―Shari Henson

Re: Systematic Review and Meta-analysis of Effects of Artichoke Extracts on Lipid Profile

Artichoke (Cynara cardunculus, Asteraceae) Cholesterol Systematic Review/Meta-analysis Date: 08-15-2017 HC# 071761-574 Sahebkar A, Pirro M, Banach M, Mikhailidis DP, Atkin SL, Cicero AFG. Lipid-lowering activity of artichoke extracts: a systematic review and meta-analysis. Crit Rev Food Sci Nutr. June 2017; [epub ahead of print]. doi: 10.1080/10408398.2017.1332572. Elevated cholesterol is a major risk factor for cardiovascular disease. To prevent and alleviate cardiovascular damage, cholesterol-lowering medication is often recommended. Although there are standard therapies for this purpose, the investigation of botanicals as adjuvant or alternative treatments might provide additional treatment options through which pharmaceutical therapies can be either delayed or avoided altogether. While there have been many studies conducted exploring the lipid-lowering effects of various botanicals, the majority of the studies are short term and small in nature. To that end, preclinical and clinical data collected indicate that artichoke (Cynara cardunculus, Asteraceae) leaf extract (ALE) has potent lipid-lowering and hepatoprotective properties. Scientists think that these properties are likely due to the plant's antioxidants, such as dicaffeoylquinic acid (cynarin and chlorogenic acid), caffeic acid, volatile sesquiterpenes, and flavonoids such as the glycosides luteolin-7-beta-rutinoside (scolymoside), luteolin-7-beta-D-glucoside, and luteolin-4-beta-D-glucoside. Finally, ALE may lower lipid levels by accelerating the excretion rate of fecal bile acids. In fact, ALE has been shown to attenuate hyperlipidemia; however, a previous meta-analysis of three randomized clinical trials revealed study limitations that may have precluded definitive conclusions about artichoke's potential efficacy. This systematic review and meta-analysis addressed whether low-density lipoprotein (LDL) cholesterol would be decreased in those taking ALE. The authors searched PubMed-Medline, Scopus, and ISI Web of Knowledge from inception to March 28, 2017. Search terms included "artichoke," "Cynara cardunculus," and "Cynara scolymus," among others. Inclusion criteria were that trials were randomized, controlled trials (parallel or crossover); investigated "artichoke products" compared with placebo and their impact on plasma or serum lipid concentrations; and measured lipid concentrations both at the start of the study and the end. Trials that were not randomized, did not include a placebo group, were observational, or did not have baseline or endpoint lipid concentrations were excluded. Data extracted from the trials were the first author's name; publication year; country in which the study was conducted; study design; total number of participants and their age, gender, and body mass index; artichoke treatment and dosage; duration; and lipid concentrations at baseline and endpoint. From a total of 66 trials, 18 were subject to inclusion and exclusion criteria, yielding nine that met the criteria for the meta-analysis. These studies contained 702 participants. Those consuming artichoke had a significant decrease in total cholesterol (weighted mean difference [WMD], −17.6 mg/dl; 95% confidence intervals [CI], −22.0, −13.3; P<0.001), LDL cholesterol (WMD, −14.9 mg/dl; 95% CI, −20.4, −9.5; P<0.011), and triglyceride concentrations (WMD, −9.2 mg/dl; 95% CI, −16.2, −2.1; P=0.011). High-density lipoprotein cholesterol was not affected by the treatment. Neither dose nor duration of artichoke supplementation was correlated with plasma LDL cholesterol concentrations. There was a significant correlation of LDL cholesterol in those consuming artichoke and concentrations at baseline (slope, −0.170; 95% CI, −0.288, 0.051; P=0.005). The Cochrane method of bias showed that all studies had low or unclear risk of bias for sequence generation, allocation concealment, selective outcome reporting, other sources of bias, blinding, and incomplete outcome data. Two statistical tests determined that there was little chance of publication bias regarding the LDL cholesterol. This meta-analysis suggests that artichoke consumption may lower total and LDL cholesterol and triglyceride concentrations. Researchers have proposed three potential mechanisms by which artichoke produces its lipid-lowering effects. For example, luteolin interferes with the activity of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, the primary enzyme responsible for controlling the rate by which the human body synthesizes its own cholesterol from its precursor mevalonate. Luteolin also interacts with liver sterol regulatory element-binding proteins (SREBPs) and acetyl-CoA C-acetyltransferase (ACAT). The authors further surmise that artichoke's lipid-lowering effects are independent of dosage and duration, and may be associated with baseline LDL concentrations. Discussed limitations include the small sample size of studies and variations in artichoke dosage and concentration. Also, both ALE and concentrated artichoke juice were used in studies, and baseline lipid concentrations were variable. This study suggests that artichoke may be worthy of further study regarding its lipid-lowering bioactivity. —Amy C. Keller, PhD

SCIENTIFIC NAME: Piper methysticum FAMILY NAME: Piperaceae COMMON NAME: kava

http://www.herbmed.org/Sponsored/kavasubcat.html

A juice company dumped orange peels in a national park. Here's what it looks like now

http://www.upworthy.com/a-juice-company-dumped-orange-peels-in-a-national-park-heres-what-it-looks-like-now?g=3&c=ufb1

Canada's 24,000 km Path Across The Country Is Now 100% Fully Connected

https://www.mtlblog.com/whats-happening/canadas-24000-km-path-across-the-country-is-now-100-fully-connected

THE PLAN TO END SCIENCE’S SEXIST #MANEL PROBLEM

sARAH SCOLES SCIENCE 08.01.1707:00 AM https://www.wired.com/story/the-plan-to-end-sciences-sexist-manel-problem/?utm_content=buffer46bff&utm_medium=social&utm_source=twitter.com&utm_campaign=buffer IN OCTOBER 2016, the organizers behind a microbiome conference sent promo materials to some prominent scientists. Elisabeth Bik was one of them. With nearly 12,000 followers, her tweets could help publicize their upcoming event in San Diego. But when she scanned the lineup, she noticed that almost every speaker was a man. Add more women, she suggested—or the conference should expect backlash. She was right: Biologist Jonathan Eisen—“Guardian of Microbial Diversity,” his Twitter bio says—brought the biased list to the attention of his 46,000 followers with a blog post called "The White Men's Microbiome Congress." The organizers, Kisaco Research, added more female speakers before the conference convened and issued a penitent statement. Bik, who runs the widely read Microbiome Digest, didn’t raise the alarm at the time. “They looked like they were going to do better," she says, "so I didn't want to make a big stink." But last week she saw the latest speaker list for the Kisaco-organized European Microbiome Congress happening this November: same story. Eisen did, too. Another white men's microbiome meeting from Kisaco #YAMMM #manel #STEMDiversity http://phylogenomics.blogspot.com/2017/07/another-white-mens-microbiome-meeting.html?spref=tw … In the past few months, two other high profile science conferences—Starmus and the World Science Festival—have also ignited internet ire for their lack of representation. And websites exist specifically to point out the most egregious examples: There’s Bias Watch Neuro, an All Male Panels Tumblr, and the hashtag #manel. The Gender Avengers, a community dedicated to hearing women's voices in public conversation, asks professionals to pledge not to serve on such panels. Yes, it's 2017. Yes, this is still happening. “Women tell themselves, ‘Our generation is going to do better. When I'm in my 40s, I'll be the speaker,’” Bik says. “I thought that. It hasn't happened.” Women have experienced underrepresentation over decades and in different departments of study. And it has real-world repercussions: Who enters science and who rises to the top of a field both have a dramatic impact on the type of research that gets done. But people like Bik, and the online communities around them, are working to make it better next time. Really. Starmussed Gender representation has been pretty imbalanced for the VIP-laden Starmus conference's six-year history, in part because organizers pride themselves on inviting fancy people of a specific sort. Nobel laureates, astronauts, Stephen Hawkings—all designations sooted with historical and cultural biases of their own. Although Starmus still doesn’t have speaker stats to boast about—less than a quarter of main-stage speakers have been women—more female scientific stars appeared at the June meeting in Trondheim, Norway, than in the past. Still, “there got to be an undercurrent in the audience, when you see this stage full of men with a token woman or no one at all," says astronomer Jill Tarter, who has been the only woman on Starmus' board. "It just festered.” The festering reached a fever pitch during a panel with seven men and zero women, after economist Christopher Pissarides confessed that he had changed Siri to a male voice. You know, because he trusts it more. When question time came, Tarter commandeered the mic. “I’m wondering,” she said, “why after a beautiful, inspiring lecture by Jeffrey Sachs this morning about [how] we have to solve our problems globally—everybody needs to be in the game—why our very wise, knighted Nobel laureate found two opportunities on the stage of this conference to piss off half the world’s population?” After the session, young women mobbed Tarter with gratitude. It's one thing to be a well-known scientist like Tarter, demanding attention at a microphone. But participants have stepped up too, as one audience member did at the World Science Festival in New York in June. There, theoretical physicist Veronika Hubeny found herself surrounded by six men, not given much opportunity to speak for the first hour. "We haven't heard enough from you," the moderator said, and started to ask her a question. But he then repeatedly talked over her to explain string theory (her field) instead of allowing her to answer. After about three minutes of intermittent interruptions from the moderator, audience member Marilee Talkington shouted: “Let her speak, please!” The room erupted into clapping and cheers—support that continued after Talkington recalled the account on Facebook. Thanks directed to Tarter multiplied online as well. But so did the thousand discriminatory papercuts from speakers and organizers. Today, the public record of sexism, at Starmus and the World Science Festival and beyond, reaches past the physical conferences and their chronology—to the postdoc watching the livestream on lunch break, to the student who searches YouTube five years from now to learn about astronauts. Instead of inspiration, they can find a demonstration of just how steep the uphill battle is. What Now? The problems on display at science conferences aren't new. To some extent, they reflect the fundamental gender imbalance in science: The tenured scientific elite has higher male-to-female ratios than the ranks of postdocs and assistant professors. But speaker imbalance still often outstrips that within a field. Self-promotion may amplify the divide: Men on average are more likely to see and sell themselves as important figures (a tendency that shows up on paper, with men citing their own work 56 percent more than women). So how do you get those numbers to change? If you talk to conference organizers, especially ones with a surfeit of men, they’ll often exclaim (as both Starmus and Microbiome Congress organizers did) that they invited more women. Those women just declined the opportunity! Here's why: They're busy. Conference organizers often have, in their heads, a list of Rock Star Female Scientists to scan through when they need some women. But those rock stars are already attending 55,000 conferences. “You have to invite more women than men because they're being stretched thin,” Bik says. The good news is that pseudocelebrity scientists aren't the only ones who do robust research and speak comprehensibly. Finding other contributors isn't hard—it just requires looking to different sources. Bik, for example, maintains a list of women in microbiology who would be happy to give a great keynote speech at your conference. The American Astronomical Society has a similar database. Organizers can also check out this Diversity Distribution Calculator to see how their meetings measure up. The field of microbiology also offers some hope. In 2011, women made up just 27 percent of the speakers at the American Society of Microbiology general meeting. By 2015, the society had bumped that up to nearly 50 percent. How? Researchers from Johns Hopkins University showed organizers numbers from their own meetings: When the committee in charge of speaker selection included at least one woman, sessions had 72 percent more female speakers and were 70 percent less likely to be only male. In response to this and other past-meeting data—and then a call to be better about avoiding all-male panels—conference conveners brought more women into the decision-making, and soon the number of women speaking nearly matched the number of men speaking. Some conferences set out with the goal of gender parity, and then choose their speaker list accordingly. It requires planning, sure, but Twitter is here to keep scientists from stalling out in their search. The key? Just ask, like neurobiologist Leslie Voshall did a few days ago as she began to schedule talks for 2019. If an organizer doesn't have enough reach of their own, they can solicit suggestions using hashtags such as #WomenInSTEM or search for lists of science-internet influencers such as the WomenTweetScienceToo rolodex, which popped up after Science put only four female scientists on its top 50 tweeters list. It has 316 badass, smart women who can write 140 informative characters or, you know, wow a weary audience at the 8 am plenary session. Coordinators can also ping #BlackInSTEM and #QueerInSTEM for speaker suggestions. Because diversity isn't just about women. “Anybody who is a minority will feel the same,” Bik says. “They will look at the podium and wish there was someone there who looked like them.” Thanks to the internet, there's really no excuse for that wish to go unfulfilled.

Full Professorship Philosophy of the Medical Sciences (U. Bordeaux, France)

The University of Bordeaux (France) hires a Full Professor in Philosophy of the Medical Sciences. The University of Bordeaux offers a thriving environment for research. It is expected that the recruited Professor will work in Thomas Pradeu's group, specialized in Philosophy of Biology, Philosophy of Medicine, and Theoretical Biology and Medicine, within ImmunoConcept, a biomedical research unit. Details: https://www.galaxie.enseignementsup-recherche.gouv.fr/ensup/ListesPostesPublies/FIDIS/0333298F/FOPC_0333298F_277.pdf Contact: Professor Jean-Francois Moreau. Application deadline: September 21th, 2017. Start date: December 31st, 2017. We would be grateful if you could circulate this announcement as widely as possible. Sincerely, Thomas Pradeu Research Director in Philosophy of Science at CNRS PI ERC project IDEM Immunology Unit, CNRS & University of Bordeaux UMR5164 146 rue Léo Saignat 33076 Bordeaux, France https://www.immuconcept.org/conceptual-immunology/ Associate Editor, Biology & Philosophy -- For questions about HOPOS-g, email the list master David Stump: davidjamesstump@gmail.com

Brominated flame retardants (BFRs) in eggs from birds of prey from Southern Germany, 2014

Environ Pollut. 2017 Aug 24;231(Pt 1):569-577. doi: 10.1016/j.envpol.2017.08.047. [Epub ahead of print] Vetter W1, Gallistl C2, Schlienz A2, Preston T2, Müller J2, von der Trenck KT3. Author information 1 University of Hohenheim, Institute of Food Chemistry, Garbenstr. 28, D-70599 Stuttgart, Germany. Electronic address: walter.vetter@uni-hohenheim.de. 2 University of Hohenheim, Institute of Food Chemistry, Garbenstr. 28, D-70599 Stuttgart, Germany. 3 Institute for the Environment, Measurements, and Nature Protection of the German State of Baden-Württemberg (LUBW), Griesbachstr. 1, D-76185 Karlsruhe, Germany. Abstract In Southern Germany, peregrine falcons (Falco peregrinus), which almost exclusively prey on other birds, are top predators of the terrestrial food chain. These animals accumulate persistent organic pollutants (POPs) and halogenated flame retardants (HFRs) with mothers transferring these lipophilic contaminants to their eggs. Here we analyzed unhatched eggs of eleven peregrine falcons and six of other species, and report concentrations of polybrominated diphenyl ethers (PBDEs), hexabromocyclododecane (HBCD), hexabromobenzene (HBB), 2,3-dibromopropyl-2,4,6-tribromophenyl ether (DPTE) and its metabolites, pentabromoethylbenzene (PBEB), pentabromotoluene (PBT), and tribromophenol (TBP). The extract of one purified peregrine falcon egg sample was comprehensively analyzed in a non-target (NT) approach by gas chromatography with mass spectrometry in the electron capture negative ion mode. A total of ∼400 polyhalogenated compounds were detected, among them dechloranes and possibly transformation products, two tetrabrominated metabolites of PBT and several compounds unknown to us which could not be identified. Copyright © 2017 Elsevier Ltd. All rights reserved. KEYWORDS: Bird eggs; Metabolites; Non-target analysis; Peregrine falcons; Polybrominated flame retardants PMID: 28843896 DOI: 10.1016/j.envpol.2017.08.047

The superior effect of nature based solutions in land management for enhancing ecosystem services

Sci Total Environ. 2017 Aug 20;610-611:997-1009. doi: 10.1016/j.scitotenv.2017.08.077. [Epub ahead of print] Keesstra S1, Nunes J2, Novara A3, Finger D4, Avelar D5, Kalantari Z6, Cerdà A7. Author information 1 Soil Physics and Land Management Group, Wageningen University, Droevendaalsesteeg 4, 6708PB Wageningen, The Netherlands; Civil, Surveying and Environmental Engineering, The University of Newcastle, Callaghan 2308, Australia. Electronic address: saskia.keesstra@wur.nl. 2 Soil Physics and Land Management Group, Wageningen University, Droevendaalsesteeg 4, 6708PB Wageningen, The Netherlands; CE3C - Centre for Ecology, Evolution and Environmental Changes, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal. Electronic address: jpcnunes@fc.ul.pt. 3 Dipartimento dei Sistemi Agro-ambientali, University of Palermo, viale delle scienze, Italy. Electronic address: agatanovara@unipa.it. 4 School of Science and Engineering. Reykjavik University, Iceland. Electronic address: davidf@ru.is. 5 CE3C - Centre for Ecology, Evolution and Environmental Changes, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal. Electronic address: dnavelar@fc.ul.pt. 6 Stockholm University, Department of Physical Geography and Bolin Centre for Climate Research, SE-106 91 Stockholm, Sweden. Electronic address: Zahra.kalantari@natgeo.su.se. 7 Soil Erosion and Degradation Research Group, Department of Geography, University of Valencia, Valencia, Spain. Electronic address: artemio.cerda@uv.es. Abstract The rehabilitation and restoration of land is a key strategy to recover services -goods and resources- ecosystems offer to the humankind. This paper reviews key examples to understand the superior effect of nature based solutions to enhance the sustainability of catchment systems by promoting desirable soil and landscape functions. The use of concepts such as connectivity and the theory of system thinking framework allowed to review coastal and river management as a guide to evaluate other strategies to achieve sustainability. In land management NBSs are not mainstream management. Through a set of case studies: organic farming in Spain; rewilding in Slovenia; land restoration in Iceland, sediment trapping in Ethiopia and wetland construction in Sweden, we show the potential of Nature based solutions (NBSs) as a cost-effective long term solution for hydrological risks and land degradation. NBSs can be divided into two main groups of strategies: soil solutions and landscape solutions. Soil solutions aim to enhance the soil health and soil functions through which local eco-system services will be maintained or restored. Landscape solutions mainly focus on the concept of connectivity. Making the landscape less connected, facilitating less rainfall to be transformed into runoff and therefore reducing flood risk, increasing soil moisture and reducing droughts and soil erosion we can achieve the sustainability. The enhanced eco-system services directly feed into the realization of the Sustainable Development Goals of the United Nations. Copyright © 2017 Elsevier B.V. All rights reserved. KEYWORDS: Ecosystem services; Nature based solutions; SDGs; System dynamics PMID: 28838037 DOI: 10.1016/j.scitotenv.2017.08.077

Micropollutants and chemical residues in organic and conventional meat

Food Chem. 2017 Oct 1;232:218-228. doi: 10.1016/j.foodchem.2017.04.013. Epub 2017 Apr 5. . Dervilly-Pinel G1, Guérin T2, Minvielle B3, Travel A4, Normand J5, Bourin M4, Royer E3, Dubreil E6, Mompelat S6, Hommet F2, Nicolas M2, Hort V2, Inthavong C2, Saint-Hilaire M2, Chafey C2, Parinet J2, Cariou R7, Marchand P7, Le Bizec B7, Verdon E6, Engel E8. Author information 1 LUNAM Université, ONIRIS, Laboratoire d'Etude des Résidus et Contaminants dans les Aliments (LABERCA), Nantes F-44307, France. Electronic address: laberca@oniris-nantes.fr. 2 Université de Paris-Est, Anses, Laboratory for Food Safety, F94700 Maisons-Alfort, France. 3 IFIP, La Motte au Vicomte, 35650 Le Rheu, France. 4 ITAVI, INRA Centre de Tours, 37380 Nouzilly, France. 5 Institut de l'Elevage, Meat Quality Department, Agrapole, 23 rue Jean Baldassini, 69364 Lyon, France. 6 ANSES - Laboratoire de Fougères, France. 7 LUNAM Université, ONIRIS, Laboratoire d'Etude des Résidus et Contaminants dans les Aliments (LABERCA), Nantes F-44307, France. 8 INRA, UR370 QuaPA, Microcontaminants, Aroma & Separation Science Group (MASS), 63123 Saint-Genes-Champanelle, France. Abstract The chemical contamination levels of both conventional and organic meats were assessed. The objective was to provide occurrence data in a context of chronic exposure. Environmental contaminants (17 polychlorinated dibenzodioxins/dibenzofurans, 18 polychlorinated biphenyls (PCBs), 3 hexabromocyclododecane (HBCD) isomers, 6 mycotoxins, 6 inorganic compounds) together with chemical residues arising from production inputs (75 antimicrobials, 10 coccidiostats and 121 pesticides) have been selected as relevant compounds. A dedicated sampling strategy, representative of the French production allowed quantification of a large sample set (n=266) including both conventional (n=139) and organic (n=127) raw meat from three animal species (bovine, porcine, poultry). While contamination levels below regulatory limits were measured in all the samples, significant differences were observed between both species and types of farming. Several environmental contaminants (Dioxins, PCBs, HBCD, Zn, Cu, Cd, Pb, As) were measured at significantly higher levels in organic samples. Copyright © 2017 Elsevier Ltd. All rights reserved. KEYWORDS: Anticoccidials; Antimicrobials; Chemical food safety; Inorganic contaminants; Mycotoxins; Organic meat; POPs; Pesticides PMID: 28490068 DOI: 10.1016/j.foodchem.2017.04.013

Reproducibility of the effects of homeopathically potentised Argentum nitricum on the growth of Lemna gibba L. in a randomised and blinded bioassay

Homeopathy. 2017 Aug;106(3):145-154. doi: 10.1016/j.homp.2017.04.001. Epub 2017 May 9. . Majewsky V1, Scherr C2, Schneider C3, Arlt SP4, Baumgartner S5. Author information 1 Institute of Complementary Medicine, University of Bern, Fabrikstrasse 8, 3012 Bern, Switzerland; Clinic for Animal Reproduction, Faculty of Veterinary Medicine, Freie Universität Berlin, Königsweg 65 (Haus 27), 14163 Berlin, Germany. Electronic address: veramajewsky@hotmail.com. 2 Institute of Complementary Medicine, University of Bern, Fabrikstrasse 8, 3012 Bern, Switzerland; Hiscia Institute, Society for Cancer Research, Kirschweg 9, 4144 Arlesheim, Switzerland. Electronic address: c.scherr@vfk.ch. 3 Research Institute of Organic Agriculture FiBL, Ackerstrasse, Postfach, 3070 Frick, Switzerland. Electronic address: claudia.schneider@fibl.org. 4 Clinic for Animal Reproduction, Faculty of Veterinary Medicine, Freie Universität Berlin, Königsweg 65 (Haus 27), 14163 Berlin, Germany. Electronic address: sebastian.arlt@fu-berlin.de. 5 Institute of Complementary Medicine, University of Bern, Fabrikstrasse 8, 3012 Bern, Switzerland; Hiscia Institute, Society for Cancer Research, Kirschweg 9, 4144 Arlesheim, Switzerland; Institute of Integrative Medicine, University of Witten-Herdecke, Gerhard-Kienle-Weg 4, 58313 Herdecke, Germany. Electronic address: stephan.baumgartner@uni-wh.de. Abstract BACKGROUND: A previous study reported a significant statistical interaction between experiment date and treatment effect of Argentum nitricum 14x-30x on the growth rate of duckweed (Lemna gibba L.). The aim of the present study was to investigate the stability of the test system and intra-laboratory reproducibility of the effects found. METHODS: Duckweed was treated with A. nitricum potencies (14x-30x) as well as succussed and unsuccussed water controls. The outcome parameter area-related growth rate for day 0-7 was determined by a computerised image analysis system in two series of independent randomised and blinded experiments. Systematic negative control (SNC) experiments were carried out to investigate test system stability. Statistical analysis was performed with full two-way analysis of variance (ANOVA) and protected Fisher's Least Significant Difference (LSD) test. RESULTS: In the first repetition series we found a significant treatment effect (p = 0.016), while in the second series no effect was observed. The negative control experiments showed that the experimental system was stable. An a posteriori subgroup analysis concerning gibbosity revealed the importance of this growth state of L. gibba for successful reproduction of the statistically significant interaction in the original study; flat: no interaction (p = 0.762); slight gibbosity: no interaction (p = 0.356); medium gibbosity: significant interaction (p = 0.031), high gibbosity: highly significant interaction (p = 0.005). CONCLUSIONS: With the original study design (disregarding gibbosity status of L. gibba) results of the original study could not be reproduced sensu stricto. We conclude that the growth state gibbosity is crucial for successful reproduction of the original study. Different physiological states of the test organisms used for bioassays for homeopathic basic research must carefully be considered. Copyright © 2017 The Faculty of Homeopathy. Published by Elsevier Ltd. All rights reserved. KEYWORDS: Duckweed; Homeopathy; Plants; Reproducibility; Silver nitrate PMID: 28844287 DOI: 10.1016/j.homp.2017.04.001

Think for yourself

Some Thoughts and Advice for Our Students and All Students August 28, 2017 https://jmp.princeton.edu/announcements/some-thoughts-and-advice-our-students-and-all-students We are scholars and teachers at Princeton, Harvard, and Yale who have some thoughts to share and advice to offer students who are headed off to colleges around the country. Our advice can be distilled to three words: Think for yourself. Now, that might sound easy. But you will find—as you may have discovered already in high school—that thinking for yourself can be a challenge. It always demands self-discipline and these days can require courage. In today’s climate, it’s all-too-easy to allow your views and outlook to be shaped by dominant opinion on your campus or in the broader academic culture. The danger any student—or faculty member—faces today is falling into the vice of conformism, yielding to groupthink. At many colleges and universities what John Stuart Mill called “the tyranny of public opinion” does more than merely discourage students from dissenting from prevailing views on moral, political, and other types of questions. It leads them to suppose that dominant views are so obviously correct that only a bigot or a crank could question them. Since no one wants to be, or be thought of, as a bigot or a crank, the easy, lazy way to proceed is simply by falling into line with campus orthodoxies. Don’t do that. Think for yourself. Thinking for yourself means questioning dominant ideas even when others insist on their being treated as unquestionable. It means deciding what one believes not by conforming to fashionable opinions, but by taking the trouble to learn and honestly consider the strongest arguments to be advanced on both or all sides of questions—including arguments for positions that others revile and want to stigmatize and against positions others seek to immunize from critical scrutiny. The love of truth and the desire to attain it should motivate you to think for yourself. The central point of a college education is to seek truth and to learn the skills and acquire the virtues necessary to be a lifelong truth-seeker. Open-mindedness, critical thinking, and debate are essential to discovering the truth. Moreover, they are our best antidotes to bigotry. Merriam-Webster’s first definition of the word “bigot” is a person “who is obstinately or intolerantly devoted to his or her own opinions and prejudices.” The only people who need fear open-minded inquiry and robust debate are the actual bigots, including those on campuses or in the broader society who seek to protect the hegemony of their opinions by claiming that to question those opinions is itself bigotry. So don’t be tyrannized by public opinion. Don’t get trapped in an echo chamber. Whether you in the end reject or embrace a view, make sure you decide where you stand by critically assessing the arguments for the competing positions. Think for yourself. Good luck to you in college! Paul Bloom Brooks and Suzanne Ragen Professor of Psychology Yale University Nicholas Christakis Sol Goldman Family Professor of Social and Natural Science Yale University Carlos Eire T. Lawrason Riggs Professor of History and Religious Studies Yale University Maria E. Garlock Professor of Civil and Environmental Engineering and Co-Director of the Program in Architecture and Engineering Princeton University Robert P. George McCormick Professor of Jurisprudence and Director of the James Madison Program in American Ideals and Institutions Princeton University Mary Ann Glendon Learned Hand Professor of Law Harvard University Joshua Katz Cotsen Professor in the Humanities and Professor of Classics Princeton University Thomas P. Kelly Professor of Philosophy Princeton University Jon Levenson Albert A. List Professor of Jewish Studies Harvard University John B. Londregan Professor of Politics and International Affairs Princeton University Michael A. Reynolds Associate Professor of Near Eastern Studies Princeton University Jacqueline C. Rivers Lecturer in Sociology and African and African-American Studies Harvard University Noël Valis Professor of Spanish Yale University Tyler VanderWeele Professor of Epidemiology and Biostatistics and Director of the Program on Integrative Knowledge and Human Flourishing Harvard University Adrian Vermeule Ralph S. Tyler, Jr. Professor of Constitutional Law Harvard University

New Osteoporosis Treatment Uses Traditional Chinese Herb To Prevent Bone Loss

MEDICINE & HEALTH By Scienmag Last updated Aug 29, 2017 An herb widely used in traditional Chinese medicine might hold the key to a new osteoporosis therapy that could prevent bone loss without causing side effects. Using a compound derived from red sage, UBC researchers have found a way to selectively block an enzyme called Cathepsin K (CatK), which plays a major role in the breakdown of collagen in bones during osteoporosis. The findings were published today in the Journal of Bone and Mineral Research. “The development of osteoporosis drugs by pharmaceutical companies has focused heavily on blocking CatK in recent years,” said Dieter Brömme, a professor in the faculty of dentistry and a Canada Research Chair in Proteases and Disease. “All clinical trials to date have failed due to side effects ranging from stroke, skin fibrosis and cardiovascular issues. We’ve found a way to block CatK only in bone tissue that we think will prevent these other negative effects.” Aug 29, 2017 The researchers tested a compound derived from red sage in human and mouse bone cells and a mouse model. They found that it prevented bone loss and increased the bone mineral density of the mice treated with the compound by 35 per cent, when compared with the control group. The study builds on previous research by Brömme and his team that looked at the effectiveness of red sage, known as Danshen in Chinese and used to treat bone ailments, in stopping the activity of CatK in limited ways. Enzyme blockers work like keys in locks. Most drugs in development have been so called active site-directed inhibitors, which act like master keys and lock the entire enzyme, blocking both its disease-relevant functions such as collagen degradation and its other normal functions. “CatK is a multifunctional enzyme with important roles in other parts of the body and we think completely blocking it is what causes unexpected side effects in other drugs,” said Preety Panwar, a research associate in the Brömme lab. “Our compound only locks the collagen -degrading CatK activity, preventing the unregulated breakdown of collagen in bones without any other negative impacts.” The treatment could also potentially be used to treat a variety of other bone and cartilage diseases such as arthritis and certain bone cancers. Osteoporosis is a global health problem that will affect one out of three women and one of out five men worldwide, with a multi-billion-dollar pharmaceutical industry dedicated to finding treatments to stop its progression. ### Media Contact Sachi Wickramasinghe sachi.wickramasinghe@ubc.ca 604-822-4636 @UBCnews http://www.ubc.ca http://dx.doi.org/10.1002/jbmr.3227

Dark Chocolate With Olive Oil Associated With Improved Cardiovascular Risk Profile

MEDICINE & HEALTH By Scienmag Last updated Aug 29, 2017 Dark chocolate enriched with extra virgin olive oil is associated with an improved cardiovascular risk profile, according to research presented today at ESC Congress.1 “A healthy diet is known to reduce the risk of cardiovascular disease,” said lead author Dr Rossella Di Stefano, a cardiologist at the University of Pisa, Italy. “Fruits and vegetables exert their protective effects through plant polyphenols, which are found in cocoa, olive oil, and apples. Research has found that the Italian Panaia red apple has very high levels of polyphenols and antioxidants.” This study tested the association between consumption of dark chocolate enriched with extra virgin olive oil or Panaia red apple (table 1) with atherosclerosis progression in healthy individuals with cardiovascular risk factors. Read More New antibodies target protein structures common to several… Aug 29, 2017 Japanese study questions benefit of treat-to-target statin therapy… Aug 29, 2017 International study shows moderate consumption of fats and… Aug 29, 2017 The randomised crossover study included 26 volunteers (14 men, 12 women) with at least three cardiovascular risk factors (smoking, dyslipidaemia, hypertension, or family history of cardiovascular disease) who received 40 grams of dark chocolate daily for 28 days. For 14 consecutive days it contained 10% extra virgin olive oil and for 14 consecutive days it contained 2.5% Panaia red apple. The two types of chocolate were given in random order. Progression of atherosclerosis was assessed by metabolic changes (levels of carnitine and hippurate), lipid profile, blood pressure and levels of circulating endothelial progenitor cells (EPCs). EPCs are critical for vascular repair and maintenance of endothelial function. Urine and blood samples were collected at baseline and after the intervention. Urine samples were analysed by proton nuclear magnetic resonance spectroscopy for endogenous metabolites. Circulating EPC levels were assessed with flow cytometry. Smoking status, body mass index, blood pressure, glycaemia and lipid profile were also monitored. After 28 days, the researchers found that the chocolate enriched with olive oil was associated with significantly increased EPC levels and decreased carnitine and hippurate levels compared to both baseline and after consumption of apple-enriched chocolate. Olive oil-enriched chocolate was associated with significantly increased high-density lipoprotein (“good”) cholesterol and decreased blood pressure compared to baseline. There was a non-significant decrease in triglyceride levels with apple-enriched chocolate. Dr Di Stefano said: “We found that small daily portions of dark chocolate with added natural polyphenols from extra virgin olive oil was associated with an improved cardiovascular risk profile. Our study suggests that extra virgin olive oil might be a good food additive to help preserve our ‘repairing cells’, the EPC.” ### Table 1: Nutrient composition of enriched dark chocolate (100 g) (70% cocoa) Media Contact ESC Press Office press@escardio.org @escardio http://www.escardio.org https://www.escardio.org/The-ESC/Press-Office/Press-releases/esc-congress-2017-hot-line-tips?hit=wireek

Exploring Alternative Use of Medicinal Plants for Sustainable Weed Management

Exploring Alternative Use of Medicinal Plants for Sustainable Weed ... www.mdpi.com/2071-1050/9/8/1468/pdf Aug 18, 2017 - Exploring Alternative Use of Medicinal Plants for. Sustainable Weed Management. Kwame Sarpong Appiah 1,2, Hossein Korrani Mardani 1,

Natural Product Research Assistant Job at Royal Botanic Gardens, Kew in London, England, United Kingdom:

https://www.researchgate.net/job/895319_Natural_Product_Research_Assistant?utm_source=twitter&utm_medium=rgShare&utm_campaign=shareJobPost via @researchgate

Interactions of Desmethoxyyangonin, a Secondary Metabolite from Renealmia alpinia , with Human Monoamine Oxidase-A and Oxidase-B

Article in Evidence-based Complementary and Alternative Medicine 2017(2):1-10 · August 2017 DOI: 10.1155/2017/4018724 · License: CC BY 4.0 1st Narayan D Chaurasiya 29.91 · University of Mississippi 2nd Francisco León 36.12 · University of Florida + 4 3rd Yuanqing Ding Last Babu Tekwani 44.32 · University of Mississippi Show more authors Abstract Renealmia alpinia (Zingiberaceae), a medicinal plant of tropical rainforests, is used to treat snakebites and other injuries and also as a febrifuge, analgesic, antiemetic, antiulcer, and anticonvulsant. The dichloromethane extract of R. alpinia leaves showed potent inhibition of human monoamine oxidases- (MAOs-) A and B. Phytochemical studies yielded six known compounds, including pinostrobin 1 , 4′-methyl ether sakuranetin 2 , sakuranetin 3 , pinostrobin chalcone 4 , yashabushidiol A 5 , and desmethoxyyangonin 6 . Compound 6 displayed about 30-fold higher affinity for MAO-B than MAO-A, with Ki values of 31 and 922 nM, respectively. Kinetic analysis of inhibition and equilibrium-dialysis dissociation assay of the enzyme-inhibitor complex showed reversible binding of desmethoxyyangonin 6 with MAO-A and MAO-B. The binding interactions of compound 6 in the active site of the MAO-A and MAO-B isoenzymes, investigated through molecular modeling algorithms, confirmed preferential binding of desmethoxyyangonin 6 with MAO-B compared to MAO-A. Selective reversible inhibitors of MAO-B, like desmethoxyyangonin 6, may have important therapeutic significance for the treatment of neurodegenerative disorders, such as Parkinson’s disease and Alzheimer’s disease Interactions of Desmethoxyyangonin, a Secondary Metabolite from Renealmia alpinia , with Human Monoamine Oxidase-A and Oxidase-B. Available from: https://www.researchgate.net/publication/319279512_Interactions_of_Desmethoxyyangonin_a_Secondary_Metabolite_from_Renealmia_alpinia_with_Human_Monoamine_Oxidase-A_and_Oxidase-B [accessed Aug 29, 2017].

Free Maillard Reaction Products in Milk Reflect Nutritional Intake of Glycated Proteins and Can Be Used to Distinguish "Organic" and "Conventionally" Produced Milk

J Agric Food Chem. 2016 Jun 22;64(24):5071-8. doi: 10.1021/acs.jafc.6b01375. Epub 2016 Jun 8. Schwarzenbolz U1, Hofmann T1, Sparmann N1, Henle T1. Author information 1 Institute of Food Chemistry, Technische Universität Dresden , D-01062 Dresden, Germany. Abstract Using LC-MS/MS and isotopically labeled standard substances, quantitation of free Maillard reaction products (MRPs), namely, N(ε)-(carboxymethyl)lysine (CML), 5-(hydroxymethyl)-1H-pyrrole-2-carbaldehyde (pyrraline, PYR), N(δ)-(5-hydro-5-methyl-4-imidazolon-2-yl)-ornithine (MG-H), and N(ε)-fructosyllysine (FL), in bovine milk was achieved. Considerable variations in the amounts of the individual MRPs were found, most likely as a consequence of the nutritional uptake of glycated proteins. When comparing commercial milk samples labeled as originating from "organic" or "conventional" farming, respectively, significant differences in the content of free PYR (organic milk, 20-300 pmol/mL; conventional milk, 400-1000 pmol/mL) were observed. An analysis of feed samples indicated that rapeseed and sugar beet are the main sources for MRPs in conventional farming. Furthermore, milk of different dairy animals (cow, buffalo, donkey, goat, ewe, mare, camel) as well as for the first time human milk was analyzed for free MRPs. The distribution of their concentrations, with FL and PYR as the most abundant in human milk and with a high individual variability, also points to a nutritional influence. As the components of concentrated feed do not belong to the natural food sources of ruminants and equidae, free MRPs in milk might serve as indicators for an adequate animal feeding in near-natural farming and can be suitable parameters to distinguish between an "organic" and "conventional" production method of milk. KEYWORDS: Maillard reaction; animal feeding; milk; organic and conventional farming; pyrraline PMID: 27213835 DOI: 10.1021/acs.jafc.6b01375 [Indexed for MEDLINE]