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The phytochemical study on the dichloromethane extract of Neolamarckia cadamba (Roxb.) Bosser has afforded two indole alkaloids, (+)-neocadambine A (1) and (-)-nauclederine (2). Their structures were confirmed by extensive spectroscopic analysis and by comparing with the reported data. (+)-Neocadambine A (1) and (-)-nauclederine (2) exhibited potent inhibition activity of advanced glycation end products (AGEs) with IC50 values of 1.2 and 0.95 mM, respectively, while the latter was almost two times more potent than the standard, aminoguanidine (1.8 mM). This is the first report on the compounds isolated from this plant with AGEs inhibition activity. In addition, (-)-nauclederine (2) was isolated for the first time in the genus of Neolamarckia. Complete 1H-NMR and 13C-NMR of compound 2 were also reported.

Thirty-three natural products were isolated from the aerial parts of Antidesma bunius, Euphorbiaceae, a plant used in Vietnamese traditional medicine against rheumatoid arthritis. All compounds were reported the first time for this species, and nine constituents resembled undescribed natural products, noticeably three coumarinolignans with 2,2-dimethyl-1,3-dioxolane moiety, two cyclopeptides, and two furofuran-type lignans connected with a phenylpropanoid moiety. The individual structures were elucidated by combining NMR and MS data, and their configuration was established by NOESY and ECD experiments and NMR calculations. Compounds with sufficient amount were analyzed for their inhibition of advanced glycation endproducts (AGEs) formation, metabolites involved in many diseases like Alzheimer, joint diseases or diabetes. With IC

This study aimed to compare the protective effect of two geographically distinct propolis against the formation of advanced glycation end products (AGEs). The polyphenol content of propolis extracts (EEPs) was analysed using HPLC‐DAD‐MS profiling and evaluated for their protective effects against pentosidine‐like AGEs. Dicarbonyl trapping capacities of major EEP compounds were determined using a methylglyoxal scavenging in vitro assay. Both propolis samples were characterised with high levels of pinobanksin derivatives exhibiting strong anti‐AGE properties. EEPs reduced AGE formation more effectively than well‐known natural AGE inhibitors such as quercetin or chlorogenic acid. Individual evaluations of the five major compounds in EEPs showed that flavonoids strongly inhibit the formation of AGEs by trapping dicarbonyl intermediates, whereas compounds such as caffeic acid derivatives only act as antioxidant agents. Propolis is thus a promising candidate for the prevention and control of AGE‐related chronic diseases.

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Background: Allergic bronchopulmonary mycosis (ABPM) is an underestimated allergic disease due to fungi. Most reported cases are caused by Aspergillus fumigatus (Af) and are referred to as allergic bronchopulmonary aspergillosis (ABPA). The main risk factor of ABPA is a history of lung disease, such as cystic fibrosis, asthma, or chronic obstructive pulmonary disease. The main diagnostic criteria for ABPA rely on the evaluation of humoral IgE and IgG responses to Af extracts, although these cannot discriminate Af sensitization and ABPA. Moreover, fungi other than Af have been incriminated. Flow cytometric evaluation of functional responses of basophils and lymphocytes in the context of allergic diseases is gaining momentum. Objectives: We hypothesized that the detection of functional responses through basophil and lymphocyte activation tests might be useful for ABPM diagnosis. We present here the results of a pilot study comparing the performance of these cellular assays vs. usual diagnostic criteria in a cystic fibrosis (CF) cohort. Methods: Ex vivo basophil activation test (BAT) is a diagnostic tool highlighting an immediate hypersensitivity mechanism against an allergen, e.g., through CD63 upregulation as an indirect measure of degranulation. Lymphocyte stimulation test (LST) relies on the upregulation of activation markers, such as CD69, after incubation with allergen(s), to explain delayed hypersensitivity. These assays were performed with Af, Penicillium, and Alternaria extracts in 29 adult CF patients. Results: BAT responses of ABPA patients were higher than those of sensitized or control CF patients. The highest LST result was for a woman who developed ABPA 3 months after the tests, despite the absence of specific IgG and IgE to Af at the time of the initial investigation. Conclusion: We conclude that basophil and lymphocyte activation tests could enhance the diagnosis of allergic mycosis, compared to usual humoral markers. Further studies with larger cohorts and addressing both mold extracts and mold relevant molecules are needed in order to confirm and extend the application of this personalized medicine approach.

Introduction La naissance prematuree est la principale cause de morbi-mortalite neonatale. Un substrat genetique, infectieux et immunologique est suspecte. Il n’existe pas actuellement de methode d’exploration non invasive pour caracteriser le statut immunitaire du nouveau-ne. Dans le cadre d’un protocole de recherche clinique sur le developpement du microbiote intestinal et le statut immunitaire dans une cohorte d’enfants prematures, cette etude ancillaire vise a proposer un dosage non invasif de mediateurs et cytokines impliques dans la physiopathologie des maladies allergiques. Methodes Apres mise au point et optimisation methodologique, le protocole experimental associe extraction proteique, concentration des selles et filtration, suivies des dosages suivants : IgE totales, tryptase (ImmunoCAP Thermo Fisher Scientific), calprotectine (BioFlash Werfen), IL-6, IL-10, IL-1 beta, TGF-beta, TNF-alpha, IgA, IgG (R&DSystems, Abcam) dans le meconium de 50 nouveau-nes prematures. Resultats Tous les mediateurs etaient detectables, a l’exception du TNF-alpha. La prevalence maximale de 100 % etait observee pour les IgA, IgG et calprotectine, suivie des IgE totales (92 %), l’IL-6 (76 %) et le TGF-beta (60 %). A l’inverse, la tryptase etait detectee dans 20 % des echantillons, l’IL-1 beta dans 30 % et l’IL-10 dans 6 % d’entre eux. La mediane des valeurs mesurees etait respectivement de 9 ng/mL (IgA), 33 ng/mL (IgG), 5 kUI/L (IgE), 514 ng/mL (calprotectine), 12 pg/mL (Il-6), 37 pg/mL (TGF-beta), 8 pg/mL (IL-1 beta), 49 pg/mL (IL-10), avec des correlations significatives tryptase-IgE totales (r = 0,99, p = 6×10–9), TGF-beta -IL-6 (r = 0,73, p = 0,0005) et TGF-beta-calprotectine (r = 0,73, p = 0,025). Discussion Nous rapportons une methodologie d’exploration immunitaire non invasive du nouveau-ne premature par analyse des mediateurs fecaux, avec des resultats dont la fourchette haute est comparable avec les valeurs disponibles dans la litterature pour des maladies intestinales allergiques et inflammatoires chroniques. Conclusion Nous apportons la preuve de concept de la faisabilite et de l’interet du dosage de mediateurs fecaux pour l’exploration non invasive de l’immunite neonatale. La perspective d’un multiplexage est discutee.

Secondary metabolites from lichens are known for exhibiting various biological effects such as anti-inflammatory, antioxidant and antibacterial activities. Despite this wide range of reported biological effects, their impact on the formation of advanced glycation end products (AGEs) remains vastly unexplored. The latter are known contributors to lifestyle and age-related diseases such as Alzheimer and Parkinson. Moreover, the development of atherosclerosis and arterial stiffness is causally linked to the formation of AGEs. With this in mind, the present work evaluated the inhibitory effects of secondary lichen metabolites on the formation of pentosidine-like AGEs' by using an in vitro, Maillard reaction based, fluorescence assay. Overall, thirty-seven natural and five synthetically modified compounds were tested, eighteen of which exhibiting IC50 values in the range of 0.05 to 0.70 mM. This corresponds to 2 to 32 fold of the inhibitory activity of aminoguanidine. Targeting one major inhibiting mechanism of AGEs formation, all compounds were additionally evaluated on their radical scavenging capacities in an DPPH assay. Furthermore, as both AGEs' formation and hypertension are major risk factors for atherosclerosis, compounds that were available in sufficient amounts were also tested for their vasodilative effects. Overall, and though some of the active compounds were previously reported cytotoxic, present results highlight the interesting potential of secondary lichen metabolites as anti-AGEs and vasodilative agents.

International audience; Advanced glycation end-products (AGEs) are associated with many pathogenic disorders such as pathogenesis of diabetes or endothelial dysfunction leading to cardiovascular events. Therefore, the identification of new anti-AGE molecules or extracts aims at preventing such pathologies. Many Clusiaceae and Calophyllaceae species are used in traditional medicines to treat arterial hypertension as well as diabetes. Focusing on these plant families, an anti-AGE plant screening allowed us to select Mammea neurophylla for further phytochemical and biological studies. Indeed, both DCM and MeOH stem bark extracts demonstrated in vitro their ability to prevent inflammation in endothelial cells and to reduce vasoconstriction. A bioguided fractionation of these extracts allowed us to point out 4-phenyl- and 4-(1-acetoxypropyl)coumarins and procyanidins as potent inhibitors of AGE formation, potentially preventing endothelial dysfunction. The fractionation steps also led to the isolation of two new compounds, namely neurophyllols A and B from the DCM bark extract together with thirteen known mammea A and E coumarins (mammea A/AA, mammea A/AB, mammea A/BA, mammea A/BB, mammea A/AA cycloD, mammea A/AB cycloD, disparinol B, mammea A/AB cycloE, ochrocarpin A, mammea A/AA cycloF, mammea A/AB cycloF, mammea E/BA, mammea E/BB) as well as δ-tocotrienol, xanthones (1-hydroxy-7-methoxyxanthone, 2-hydroxyxanthone) and triterpenes (friedelin and betulinic acid). During this study, R,S-asperphenamate, previously described from fungal origin was also purified.

Introduction Les micropuces mesurant les reponses IgE et IgG specifiques vis-a-vis de > 100 allergenes sont de plus en plus utilisees a des fins diagnostiques et epidemiologiques. La miniaturisation necessite des quantites tres faibles de chaque allergene, exposant aux interferences par competition entre les IgE et les IgG. L’effet de ces interferences sur la reactivite IgE des patients est inconnu. Methodes Nous avons utilise du serum de neuf patients allergiques pour comparer la reactivite IgE observee sur micropuce ISAC (ThermoFisher, Uppsala, Suede) avec le serum natif et apres depletion des IgG (proteine G). La concentration des IgE totales (tIgE) et d’un analyte neutre ont ete mesurees dans chaque echantillon, natif et deplete. L’analyse des resultats a ete qualitative (changement de classe d’intensite, definition des classes DOI 10.1111/all.13548) et quantitative (calcul du ratio d’intensite ISU-E apres/avant depletion). L’analyse a pris en compte l’incertitude de mesure de 25 % de la micropuce. Resultats La depletion en IgG de chaque serum a conduit a une diminution du niveau de tIgE (mediane 65 % de la concentration de chaque serum natif, extremes 50 et 73 %). Un total de 225 reactivites IgE > 0,3 ISU-E etaient observees dans les sera depletes en IgG, apres correction du niveau de tIgE. Un deplacement vers des classes d’intensite superieures etait observe dans 90 cas (40 %), dont 11 reactivites IgE demasquees apres la depletion des IgG. En analyse quantitative, l’augmentation moyenne de l’intensite de reactivite chez un sujet donne variait de 1,7 a 6,9. Nous n’avons pas observe d’association privilegiee entre une forte progression de la reactivite IgE apres depletion des IgG et les proprietes des allergenes concernes. Discussion La fixation des IgG avec competition IgG-IgE etait presente chez les neuf patients de notre etude, avec comme corollaire une sous-estimation, voire l’absence de detection, de la reactivite IgE. Conclusion Nos resultats suggerent (1) les resultats de la mesure des reactivites IgE sur micropuce devraient etre interpretes avec prudence et (2) une etape de depletion des IgG prealablement a la mesure des reactivites IgE permettrait d’obtenir des resultats plus justes.

International audience; Propolis is a complex mixture used by bees to seal off hives, or use as a chemical weapon against intruders. Propolis is mainly composed of plant resins and beeswax so that its chemical composition, and consequently biological activity, varies with collection sites. Therefore propolis is generally classified as "poplar-type" in temperate zones vs "green Brazilian", "Clusia", "Macaranga" or Mediterranean-type in tropical zones [1]. The antiglycation potential of an organic poplar-type propolis sample had been already evaluated by our team. This study revealed that a DCM extract exhibited a strong anti-AGEs activity (IC50 28 µg/mL vs 90 µg/mL for the reference i.e. an EtOH extract of Styphnolobium japonicum) [2]. A bioassay-guided fractionation highlighted the major anti-AGEs components of this extract as pinobanksine derivatives and prenyl cafeate. The present workaims to show that the associated inhibition mechanism is directly related to their trapping ability of reactive dicarbonyl species such as methylglyoxal, an intermediate component in AGEs formation (Figure 1).Fig. 1. The Maillard reaction - Schematic formation of AGEs.Rapid identification of chemical markers is an important issue in propolis studies. A fast dereplication analysis of the propolis DCM extract, using a Laser Desorption Ionization (LDI) MS technique [3], allowed us to instantly identify 25 polyphenol derivatives previously identified by classical methods [2,4]. The results clearly show that LDI-MS represents a fast and powerful method to characterize propolis extracts and identify their origin.References:[1] Salatino A, Fernandes-Silva CC, Righi AA, Salatino MLF. Propolis research and the chemistry of plant products. Nat Prod Rep 2011; 28: 925–936[2] Boisard S, Le Ray A-M, Gatto J, Aumond M-C, Blanchard P, Derbré S, Flurin C, Richomme P. Chemical Composition, Antioxidant and Anti-AGEs Activities of a French Poplar Type Propolis. J Agric Food Chem 2014; 62: 1344–1351[3] Le Pogam P, Schinkovitz A, Legouin B, Le Lamer A-C, Boustie J, Richomme P. Matrix-​Free UV-​Laser Desorption Ionization Mass Spectrometry as a Versatile Approach for Accelerating Dereplication Studies on Lichens. Anal Chem 2015; 87: 10421-8[4] Boisard S, Le Ray A-M, Landreau A, Kempf M, Cassisa V, Flurin C, Richomme P. Antifungal and Antibacterial Metabolites from a French Poplar Type Propolis. Evid Based Complement Alternat Med 2015; e319240.

Autosomal dominant type IIa hypercholesterolaemia (ADH) is characterised by an elevation of total plasma cholesterol associated with increased LDL particles. Numerous different molecular defects have been identified in the LDL receptor (LDLR) and few specific mutations in the apolipoprotein B (APOB) gene resulting in familial hypercholesterolaemia and familial defective apoB-100 respectively. To estimate the respective contribution of LDLR, APOB and other gene defects in this disease, we studied 33 well characterised French families diagnosed over at least three generations with ADH through the candidate gene approach. An estimation of the proportions performed with the HOMOG3R program showed that an LDLR gene defect was involved in approximately 50% of the families (P = 0.001). On the other hand, the estimated contribution of an APOB gene defect was only 15%. This low estimation of ADH due to an APOB gene defect is further strengthened by the existence of only two probands carrying the APOB (R3500Q) mutation in the sample. More importantly and surprisingly, 35% of the families in the sample were estimated to be linked to neither LDLR nor APOB genes. These data were confirmed by the exclusion of both genes through direct haplotyping in three families. Our results demonstrate that the relative contributions of LDLR and APOB gene defects to the disease are very different. Furthermore, our results also show that genetic heterogeneity is, generally, underestimated in ADH, and that at least three major groups of defects are involved. At this point, the contribution of the recently mapped FH3 gene to ADH cannot be assessed nor its importance in the group of 'non LDLR/non APOB' families.

International audience; Accumulation in tissues and serum of advanced glycation end-products (AGEs) plays an important role in pathologies such as Alzheimer’s disease or, in the event of complications of diabetes, atherosclerosis or renal failure. Therefore, there is a potential therapeutic interest in compounds able to lower intra and extracellular levels of AGEs. Among them, natural antioxidants (AO) with true anti-AGEs capabilities would represent good candidates for development. The purpose of this study was to evaluate the AO and anti-AGEs potential of a propolis batch and then to identify the main compounds responsible for these effects. In vivo, protein glycation and oxidative stress are closely related. Thus, AO and antiglycation activities were evaluated using both DPPH and ORAC assays, respectively, as well as a newly developed automated anti-AGEs test. Several propolis extracts exhibited very good AO and anti-AGEs activities, and a bioguided fractionation allowed us to identify pinobanksin-3-acetate as the most active component.

Advanced glycation end-products (AGEs) are involved in the pathogenesis of numerous diseases. Among them, cellular accumulation of AGEs contributes to vascular complications in diabetes. Besides using drugs to lower blood sugar, a balanced diet and the intake of herbal products potentially limiting AGE formation could be considered beneficial for patients' health. The current paper presents a simple and cheap high-throughput screening (HTS) assay based on AGE fluorescence and suitable for plant extract screening. We have already implemented an HTS assay based on vesperlysines-like fluorescing AGEs quickly (24 h) formed from BSA and ribose under physiological conditions. However, interference was noted when fluorescent compounds and/or complex mixtures were tested. To overcome these problems and apply this HTS assay to plant extracts, we developed a technique for systematic quantification of both vesperlysines (λ(exc) 370 nm; λ(em) 440 nm) and pentosidine-like (λ(exc) 335 nm; λ(em) 385 nm) AGEs. In a batch of medicinal and food plant extracts, hits were selected as soon as fluorescence decreased under a fixed threshold for at least one wavelength. Hits revealed during this study appeared to contain well-known and powerful anti-AGE substances, thus demonstrating the suitability of this assay for screening crude extracts (0.1 mg/mL). Finally, quercetin was found to be a more powerful reference compound than aminoguanidine in such assay.

Background: Formation and accumulation of advanced glycation end-products (AGE) is recognized as a major pathogenic process in diabetic complications, atherosclerosis and cardiovascular diseases. In addition, reactive oxygen species and free radicals have also been reported to participate in AGE formation and in cell damage. Natural products with antioxidant and antiAGE activity have great therapeutic potential in the treatment of diabetes, hypertension and related complications. Objective: to test ethanolic extracts and aqueous-traditional preparations of plants used to treat diabetes, hypertension and obesity in Yucatecan traditional medicine for their anti-AGE and free radical scavenging activities. Materials and Methods: ethanolic extracts of leaves, stems and roots of nine medicinal plants, together with their traditional preparations, were prepared and tested for their anti-AGE and antioxidant activities using the inhibition of advanced glycation end products and DPPH radical scavenging assays, respectively. Results: the root extract of C. fistula (IC50= 0.1 mg/mL) and the leaf extract of P. auritum (IC50= 0.35 mg/mL) presented significant activity against vesperlysine and pentosidine-like AGE. Although none of the aqueous traditional preparations showed significant activity in the anti-AGE assay, both the traditional preparations and the ethanolic extracts of E. tinifolia, M. zapota, O. campechianum and P. auritum showed significant activity in the DPPH reduction assay. Conclusions: the results suggest that the metabolites responsible for the detected radical-scavenging activity are different to those involved in inhibiting AGE formation; however, the extracts with antioxidant activity may contain other metabolites which are able to prevent AGE formation through a different mechanism. SUMMARY Ethanolic extracts from nine plants used to treat diabetes, hypertension and obesity in Yucatecan traditional medicine were tested for their anti-AGE and free radical scavenging activities.Significant activity against vesperlysine and pentosidine-like AGE was detected in the root extract of Cassia fistula and the leaf extract of Piper auritum.Traditional preparations and the ethanolic extracts of Ehretia tinifolia, Manilkara zapota, Ocimum campechianum and Piper auritum showed significant activity in the DPPH reduction assay.Results suggest that the metabolites responsible for the detected radical-scavenging activity are different to those involved in inhibiting AGE formation. Abbreviations Used: AGE: Advanced glycation end-product; DPPH: 2,2-Diphenyl-1-picrylhydrazyl; DM: Diabetes mellitus; ROS: Reactive oxygen species; BSA: Bovine serum albumin; EtOH: Ethanol; EtOAc: Ethyl acetate; ANOVA: Analysis of variance; BA: Brosimum alicastrum; BS: Bunchosia swartziana; CF: Cassia fistula; CN: Cocos nucifera; ET: Ehretia tinifolia; MZ: Manilkara zapota; OC: Ocimum campechianum; PA: Piper auritum; RM: Rhizophora mangle; L: Leaves; S: Stems; R: Roots; T: traditional preparation; I: Inflorescences; W: Water

Eight clomiphene citrate (150 mg/day for 5 days)-resistant anovulatory women with polycystic ovary were included in this study. A luteinizing hormone-releasing hormone (LH-RH) analog, D-Trp-6-LH-RH, 100 micrograms subcutaneous-per day, induced a hypogonadotropic state within varying periods but at most within 3 weeks, after an initial flare-up effect characterized by slight increase in ovarian size in four patients and in the other four by cysts that disappeared rapidly. On the 28th day or 15 to 20 days after menstruation for subsequent cycles, during maintenance of D-Trp-6-LH-RH therapy, a usual gonadotropin regimen was carried out in 33 cycles. Human menopausal gonadotropins obtained follicular maturation in all cycles. However, there was never the growth of a single dominant follicle but always of several follicles. Human chorionic gonadotropin then induced ovulation in 31 cycles (94%). Luteal phase was normal in 28 and inadequate in 3 of the 31 ovulatory cycles. Hyperstimulation, generally mild to moderate but rather severe in 2 cycles, was constant. Five pregnancies were obtained. The overall pregnancy rate was 15% per cycle and 17.8% per normoovulatory cycle. This study showed that an associated treatment with an LH-RH analog enables gonadotropins to achieve ovulation regularly with an encouraging number of pregnancies but at a risk of hyperstimulation.

An adult male Callicebus moloch was presented for acute congestive heart failure. Therapy was unsuccessful and necropsy showed severe systemic atherosclerosis. Analysis of serum revealed hypercholesterolemia with specific elevation of the betalipoprotein fraction.

Après avoir rappelé la démarche de l’escalade thérapeutique chez le patient diabétique de type 2, l’auteur fait le suivi de la prescription des analogues du GLP-1 afin d’établir des critères prédictifs de leur efficacité et de discuter de l’influence de l’insulinothérapie. L’auteur présente une étude rétrospective de 86 patients diabétiques de type 2 et discute les résultats obtenus. L’ensemble des résultats ne semble pas démontrer de critère prédictif d’efficacité particulière par rapport à la littérature déjà existante. Les résultats concernant l’insulinothérapie montrent quelques bonnes évolutions et sont en faveur d’une remise en question des traitements en fonction de l’évolution du diabète. L’auteur conclut enfin en discutant de l’intérêt d’une approche centrée sur le patient dans le but d’améliorer sa prise en charge et son observance