Combination of gas chromatography–mass spectrometry and mass spectral deconvolution for structural elucidation of an unusual C29-steroid detected in a complex sedimentary matrix

A complex sedimentary sample from the Monterey Formation (CA, USA) has been submitted to GC–MS analysis followed by mass spectral deconvolution using Automated Mass Spectral Deconvolution and Identification System (AMDIS). Adjusting the parameters of the software allowed for the extraction of the spectrum of an unusual steroidal hydrocarbon coeluting with the major compound of the chromatogram. Following a careful interpretation of the "extracted" mass spectrum, the structure of the unknown has been postulated to be the 4,14-dimethylcholestane (DMC). Possible origins of this rare steroid are briefly discussed. Thus, application of AMDIS appears to be particularly suitable for the GC–MS analysis of natural complex mixtures characterized by a high number of analytes present in low amounts

Cytotoxic effect of Agaricus bisporus and Lactarius rufus β-d-glucans on HepG2 cells

AbstractThe cytotoxic activity of β-d-glucans isolated from Agaricus bisporus and Lactarius rufus fruiting bodies was evaluated on human hepatocellular carcinoma cells (HepG2). NMR and methylation analysis suggest that these β-d-glucans were composed of a linear (1→6)-linked and a branched (1→3), (1→6)-linked backbone, respectively. They both decreased cell viability at concentrations of up to 100μgmL−1, as shown by MTT assay. The amount of LDH released and the analysis of cell morphology corroborated these values and also showed that the β-d-glucan of L. rufus was more cytotoxic to HepG2 cells than that of A. bisporus. The treatment of HepG2 cells with L. rufus and A. bisporus β-d-glucans at a dose of 200μgmL−1 for 24h promoted an increase of cytochrome c release and a decrease of ATP content, suggesting that these polysaccharides could promote cell death by apoptosis. Both β-d-glucans were tested against murine primary hepatocytes at a dose of 200μgmL−1. The results suggest that the L. rufus β-d-glucan was as cytotoxic for hepatocytes as for HepG2 cells, whereas the A. bisporus β-d-glucan, under the same conditions, was cytotoxic only for HepG2 cells, suggesting cell selectivity. These results open new possibilities for use of mushroom β-d-glucans in cancer therapy

Identification of a Rare Coding Variant in Complement 3 Associated with Age-related Macular Degeneration

Macular degeneration is a common cause of blindness in the elderly. To identify rare coding variants associated with a large increase in risk of age-related macular degeneration (AMD), we sequenced 2,335 cases and 789 controls in 10 candidate loci (57 genes). To increase power, we augmented our control set with ancestry-matched exome sequenced controls. An analysis of coding variation in 2,268 AMD cases and 2,268 ancestry matched controls revealed two large-effect rare variants; previously described R1210C in the CFH gene (fcase = 0.51%, fcontrol = 0.02%, OR = 23.11), and newly identified K155Q in the C3 gene (fcase = 1.06%, fcontrol = 0.39%, OR = 2.68). The variants suggest decreased inhibition of C3 by Factor H, resulting in increased activation of the alternative complement pathway, as a key component of disease biology

A large genome-wide association study of age-related macular degeneration highlights contributions of rare and common variants.

This is the author accepted manuscript. The final version is available from Nature Publishing Group via http://dx.doi.org/10.1038/ng.3448Advanced age-related macular degeneration (AMD) is the leading cause of blindness in the elderly, with limited therapeutic options. Here we report on a study of >12 million variants, including 163,714 directly genotyped, mostly rare, protein-altering variants. Analyzing 16,144 patients and 17,832 controls, we identify 52 independently associated common and rare variants (P < 5 × 10(-8)) distributed across 34 loci. Although wet and dry AMD subtypes exhibit predominantly shared genetics, we identify the first genetic association signal specific to wet AMD, near MMP9 (difference P value = 4.1 × 10(-10)). Very rare coding variants (frequency <0.1%) in CFH, CFI and TIMP3 suggest causal roles for these genes, as does a splice variant in SLC16A8. Our results support the hypothesis that rare coding variants can pinpoint causal genes within known genetic loci and illustrate that applying the approach systematically to detect new loci requires extremely large sample sizes.We thank all participants of all the studies included for enabling this research by their participation in these studies. Computer resources for this project have been provided by the high-performance computing centers of the University of Michigan and the University of Regensburg. Group-specific acknowledgments can be found in the Supplementary Note. The Center for Inherited Diseases Research (CIDR) Program contract number is HHSN268201200008I. This and the main consortium work were predominantly funded by 1X01HG006934-01 to G.R.A. and R01 EY022310 to J.L.H

Effet de l'hypoxie sur les cellules souches mésenchymateuses de la pulpe dentaire dans un objectif d'ingénierie tissulaire

The tooth is a living organ, faced throughout life to multiple attacks (caries, trauma...) which can cause necrosis of the pulp. The development of a "pulp equivalent" could be an innovative therapeutic approach as an alternative to current endodontic treatments. The pulp of deciduous teeth is a reservoir of mesenchymal stem cells (Stem cells SHED from Human Exfoliated Deciduous teeth) with a high potential of proliferation and differentiation. The overall objective of this work was to reconstitute a functional pulp tissue by developing a pulp equivalent (pulp mesenchymal cells seeded in a 3D collagen matrix) to be grafted within the previously hollowed pulp chamber to maintain tooth vitality. The specific objectives were: In vitro: 1) to study the angiogenic potential of SHED compared with dermal fibroblasts in normoxic and hypoxic conditions. 2) to determine the optimal hypoxic preconditioning period to stimulate the angiogenic potential of SHED, 3) to identify a potential cytokine activating the capillary formation, 4) to analyze the effect of hypoxia on the expression of markers surfaces SHED, and 5) to check that hypoxia did not alter the mineralization potential of these cells. In vivo: 1) to evaluate, in a pre-clinical model of pulp equivalent implantation in ectopic site in mice, the effect of either hypoxic or FGF preconditioning on the angiogenic potential of SHED. These experiments were first conducted with mouse pulp cells and further confirmed with SHED implanted in immunodeficient mice, and 2) to develop dynamic imaging techniques to monitor neoangiogenesis within pulp equivalent. Finally, in an objective of transfer to the human dental clinic, we studied the effect of a new biomaterial based on tricalcium on tissue repair in a pulp injury model in rats, compared to gold standard materials.La dent est un tissu vivant, confronté tout au long de la vie à de multiples agressions (caries, traumatismes...) qui peuvent entraîner la nécrose de la pulpe. La mise au point d’une «pulpe équivalente» pourrait constituer une approche thérapeutique innovante comme alternative aux traitements actuels d’endodontie. La pulpe des dents temporaires constitue un réservoir de cellules souches mésenchymateuses (SHED Stem cells from Human Exfoliated Deciduous teeth) aux potentiels de prolifération et de différenciation élevés. L’objectif global de ce travail est de reconstituer un tissu pulpaire fonctionnel en développant une pulpe équivalente (cellules pulpaires mésenchymateuses ensemencées dans une matrice 3D de collagène) pour être greffée à l’intérieur de la chambre pulpaire préalablement évidée afin de conserver la vitalité de la dent. Les objectifs spécifiques ont été : In vitro : 1) d'étudier le potentiel angiogénique des SHED comparés à des fibroblastes dermiques en conditions normoxiques et hypoxiques, 2) de déterminer la durée de pré-conditionnement hypoxique optimale pour stimuler le potentiel angiogénique des SHED, 3) de sélectionner une potentielle cytokine activant la formation de capillaires, 4) d’analyser l’effet de l’hypoxie sur l’expression des marqueurs de surfaces des SHED, et 5) de vérifier que l’hypoxie n’altérait pas le potentiel de minéralisation de ces cellules. In vivo : 1) d’évaluer, dans un modèle pré-clinique d’implantation de pulpes équivalentes en site ectopique chez la souris, l’effet du pré-conditionnement hypoxique sur le potentiel angiogénique des SHED. Ces expériences ont d’abord été conduites avec des cellules pulpaires de souris puis confirmées avec des SHED implantées dans des souris immunodéficientes, et 2) de développer des techniques d’imagerie dynamique pour suivre la néoangiogenèse dans les pulpes équivalentes implantées. Enfin, dans un objectif de transfert vers la clinique dentaire humaine, nous avons étudié l’effet d’un nouveau biomatériau à base de calcium tricalcique sur la réparation tissulaire dans un modèle de blessure pulpaire chez le rat, en comparaison aux matériaux de référence

Effect of hypoxia on dental pulp mesenchymal stem cells for pulp tissue engineering

La dent est un tissu vivant, confronté tout au long de la vie à de multiples agressions (caries, traumatismes...) qui peuvent entraîner la nécrose de la pulpe. La mise au point d’une «pulpe équivalente» pourrait constituer une approche thérapeutique innovante comme alternative aux traitements actuels d’endodontie. La pulpe des dents temporaires constitue un réservoir de cellules souches mésenchymateuses (SHED Stem cells from Human Exfoliated Deciduous teeth) aux potentiels de prolifération et de différenciation élevés. L’objectif global de ce travail est de reconstituer un tissu pulpaire fonctionnel en développant une pulpe équivalente (cellules pulpaires mésenchymateuses ensemencées dans une matrice 3D de collagène) pour être greffée à l’intérieur de la chambre pulpaire préalablement évidée afin de conserver la vitalité de la dent. Les objectifs spécifiques ont été : In vitro : 1) d'étudier le potentiel angiogénique des SHED comparés à des fibroblastes dermiques en conditions normoxiques et hypoxiques, 2) de déterminer la durée de pré-conditionnement hypoxique optimale pour stimuler le potentiel angiogénique des SHED, 3) de sélectionner une potentielle cytokine activant la formation de capillaires, 4) d’analyser l’effet de l’hypoxie sur l’expression des marqueurs de surfaces des SHED, et 5) de vérifier que l’hypoxie n’altérait pas le potentiel de minéralisation de ces cellules. In vivo : 1) d’évaluer, dans un modèle pré-clinique d’implantation de pulpes équivalentes en site ectopique chez la souris, l’effet du pré-conditionnement hypoxique sur le potentiel angiogénique des SHED. Ces expériences ont d’abord été conduites avec des cellules pulpaires de souris puis confirmées avec des SHED implantées dans des souris immunodéficientes, et 2) de développer des techniques d’imagerie dynamique pour suivre la néoangiogenèse dans les pulpes équivalentes implantées. Enfin, dans un objectif de transfert vers la clinique dentaire humaine, nous avons étudié l’effet d’un nouveau biomatériau à base de calcium tricalcique sur la réparation tissulaire dans un modèle de blessure pulpaire chez le rat, en comparaison aux matériaux de référence.The tooth is a living organ, faced throughout life to multiple attacks (caries, trauma ...) which can cause necrosis of the pulp. The development of a" pulp equivalent " could be an innovative therapeutic approach as an alternative to current endodontic treatments. The pulp of deciduous teeth is a reservoir of mesenchymal stem cells (Stem cells SHED from Human Exfoliated Deciduous teeth) with a high potential of proliferation and differentiation. The overall objective of this work was to reconstitute a functional pulp tissue by developing a pulp equivalent (pulp mesenchymal cells seeded in a 3D collagen matrix) to be grafted within the previously hollowed pulp chamber to maintain tooth vitality. The specific objectives were: In vitro: 1) to study the angiogenic potential of SHED compared with dermal fibroblasts in normoxic and hypoxic conditions. 2) to determine the optimal hypoxic preconditioning period to stimulate the angiogenic potential of SHED, 3) to identify a potential cytokine activating the capillary formation, 4) to analyze the effect of hypoxia on the expression of markers surfaces SHED, and 5) to check that hypoxia did not alter the mineralization potential of these cells. In vivo: 1) to evaluate, in a pre-clinical model of pulp equivalent implantation in ectopic site in mice, the effect of either hypoxic or FGF preconditioning on the angiogenic potential of SHED. These experiments were first conducted with mouse pulp cells and further confirmed with SHED implanted in immunodeficient mice, and 2) to develop dynamic imaging techniques to monitor neoangiogenesis within pulp equivalent. Finally, in an objective of transfer to the human dental clinic, we studied the effect of a new biomaterial based on tricalcium on tissue repair in a pulp injury model in rats, compared to gold standard materials

The Miocene New Jersey Passive Margin as a Model for the Distribution of Sedimentary Organic Matter in Siliciclastic Deposits

The New Jersey margin is a classic example of a siliciclastic passive margin which prograded during the Miocene and Pliocene, forming well-defined clinoforms on seismic data. It has been explored by the petroleum industry since the 1970s and more recently by Deep Sea Drilling Program and Ocean Drilling Program legs. Within this well-constrained geological framework, the stratigraphic and spatial distribution of sedimentary organic matter (OM) in fine-grained sediments has been analyzed along a transect from paleoshelf to paleoslope provided by four ODP Sites. This led to the establishment of a depositional model for sedimentary OM in prograding clinoforms, which can be applied worldwide to other similar environments of deposition. The most significant factor influencing the type of OM sedimentation (palynofacies) is the location of the sites with respect to the shelf break. The correlation of palynofacies parameters with gamma-ray, Rock-Eval pyrolysis, and seismic data has permitted the establishment of a model for the sedimentation of OM across the margin. Various parameters indicative of proximal–distal trends have been tested, their validity tending to deteriorate distally. The most reliable ones are: (1) the continental/marine ratio (based on palynomorphs), (2) the relative percentage of amorphous OM, and (3) phytoclasts. The variations in the continental/marine ratio permits the identification of stacked transgressive–regressive intervals related to accommodation cycles and of associated condensed intervals. This palynofacies parameter can be correlated with the seismic facies at the locations of the sites. Total organic carbon (TOC) content usually varies similarly to the continental/marine ratio, which implies that most sedimentary OM is of continental origin. The only exception is in transgressive intervals, where it varies in the opposite way, thereby indicating a change in the source of sediment and an increased concentration of marine OM. Similarly, the gamma-ray log is a good proxy for proximal–distal trends (coarse to fine sediments) at all sites, where it varies similarly to the continental/marine ratio, except in condensed intervals in the distal part of clinoforms where it records the settling of uranium-rich clay particles. Furthermore, the positive correlation between the gamma-ray data, TOC data and the continental/marine ratio confirms that the hydrodynamic behavior of continental sedimentary OM is close to that of clay and silt particles. This research illustrates the contribution of sedimentary OM to the study of fine-grained sediments prograding on a passive margin. Some parameters, especially the continental/marine ratio, can be used as proxies for the sedimentological interpretation of sites and for distinguishing transgressive–regressive trends within prograding clinoforms

Evaluation of Retention, Wear, and Maintenance of Attachment Systems for Single- or Two-Implant-Retained Mandibular Overdentures: A Systematic Review

International audienceAttachment systems (AS) enhance retention and stability by anchoring the overdentures to implants. Since 2002, the McGill consensus statement recommends the 2-implant-retained overdentures as the standard choice for edentulous mandible (2-IRMO). Considering the large number of AS available, it remains difficult for a practitioner to make a reasoned choice. A systematic review was conducted in PubMed/Medline and carried out independently by three authors, on retention, wear, and maintenance of AS used clinically or in vitro specifically for 1- or 2-IRMO. The 45 selected studies include 14 clinical and 31 in vitro studies. The risk of bias was evaluated according to the revised Cochrane risk of bias tool for randomized trials (RoB 2). The initial retention force of the cylindrical system is higher than the ball system. The retention loss, related to the wear of the retention device, is responsible for the most common need of maintenance, requiring activation or replacement. Plastic retention devices wear out faster and more significantly than metal ones, implying a worse time behavior of cylindrical systems, but their maintenance rate is similar. Neither system appears categorically superior. Cylindrical systems provide higher initial retention than ball ones; this advantage reduces over time with wear without affecting their need for maintenance