Pomegranate Peel Extract Prevents Bone Loss in a Preclinical Model of Osteoporosis and Stimulates Osteoblastic Differentiation in Vitro

The nutritional benefits of pomegranate have attracted great scientific interest. The pomegranate, including the pomegranate peel, has been used worldwide for many years as a fruit with medicinal activity, mostly antioxidant properties. Among chronic diseases, osteoporosis, which is associated with bone remodelling impairment leading to progressive bone loss, could eventually benefit from antioxidant compounds because of the involvement of oxidative stress in the pathogenesis of osteopenia. In this study, with in vivo and ex vivo experiments, we investigated whether the consumption of pomegranate peel extract (PGPE) could limit the process of osteopenia. We demonstrated that in ovariectomized (OVX) C57BL/6J mice, PGPE consumption was able to significantly prevent the decrease in bone mineral density (-31.9%; p < 0.001 vs. OVX mice) and bone microarchitecture impairment. Moreover, the exposure of RAW264.7 cells to serum harvested from mice that had been given a PGPE-enriched diet elicited reduced osteoclast differentiation and bone resorption, as shown by the inhibition of the major osteoclast markers. In addition, PGPE appeared to substantially stimulate osteoblastic MC3T3-E1 alkaline phosphatase (ALP) activity at day 7, mineralization at day 21 and the transcription level of osteogenic markers. PGPE may be effective in preventing the bone loss associated with ovariectomy in mice, and offers a promising alternative for the nutritional management of this disease

Evaluation of criteria to assist the selection of good quality grafted grapevines prior to their commercialisation

The production of grafted grapevine plant material is a complex process with many steps running over at least one year, from grafting to final sorting in nurseries. To reach the market in France, grafted grapevines must meet three criteria by law: resistance to a manual graft union test (or thumb test), a minimum number of three roots and a woody, lignified stem which has grown from the bud of the scion wood of at least 2 cm long. This study aimed to evaluate the possibility of using visual criteria to select good quality grafted grapevines, without the need to do the thumb test because the thumb test is manual and therefore very subjective; the test depends on the strength applied by the person who realises it. This study was done on 22 scion/rootstock combinations with different degrees of grafting success, i.e., producing different proportions of marketable plants after one year in the nursery. The three legal criteria currently used to select marketable grafted grapevines in France as well as other external and measurable criteria such as the length of lignified stem and diameter, the number of thin and thick roots, and rootstock wood diameter were measured on the 22 scion/rootstock combinations. Variation in the values for these different criteria was observed and correlations between the criteria and the number of marketable plants were studied. This data was then analysed to determine which visible criteria contribute most to identifying marketable grafts. The percentage of marketable grafts was most strongly correlated with the thumb test and positively correlated with the length of the lignified stem. The variables with the highest predictive effect for identifying marketable plants (other than the thumb test) were the number of large roots and the length of the lignified stem. The possibility of using visible criteria to screen for good quality grafted plants is discussed, but no single, or combination of criteria, was sufficiently strongly correlated with the percentage of marketable plants to replace the thumb test

Effects of elevated CO2 on phytoplankton community biomass and species composition during a spring Phaeocystis spp. bloom in the western English Channel

A 21-year time series of phytoplankton community structure was analysed in relation to Phaeocystis spp. to elucidate its contribution to the annual carbon budget at station L4 in the western English Channel (WEC). Between 1993–2014 Phaeocystis spp. contributed ∼4.6% of the annual phytoplankton carbon and during the March − May spring bloom, the mean Phaeocystis spp. biomass constituted 17% with a maximal contribution of 47% in 2001. Upper maximal weekly values above the time series mean ranged from 63 to 82% of the total phytoplankton carbon (∼42–137 mg carbon (C) m −3 ) with significant inter-annual variability in Phaeocystis spp. Maximal biomass usually occurred by the end of April, although in some cases as early as mid-April (2007) and as late as late May (2013). The effects of elevated pCO 2 on the Phaeocystis spp. spring bloom were investigated during a fifteen-day semi-continuous microcosm experiment. The phytoplankton community biomass was estimated at ∼160 mg C m −3 and was dominated by nanophytoplankton (40%, excluding Phaeocystis spp.), Phaeocystis spp. (30%) and cryptophytes (12%). The smaller fraction of the community biomass comprised picophytoplankton (9%), coccolithophores (3%), Synechococcus (3%), dinoflagellates (1.5%), ciliates (1%) and diatoms (0.5%). Over the experimental period, total biomass increased significantly by 90% to ∼305 mg C m −3 in the high CO 2 treatment while the ambient pCO 2 control showed no net gains. Phaeocystis spp. exhibited the greatest response to the high CO 2 treatment, increasing by 330%, from ∼50 mg C m −3 to over 200 mg C m −3 and contributing ∼70% of the total biomass. Taken together, the results of our microcosm experiment and analysis of the time series suggest that a future high CO 2 scenario may favour dominance of Phaeocystis spp. during the spring bloom. This has significant implications for the formation of hypoxic zones and the alteration of food web structure including inhibitory feeding effects and lowered fecundity in many copepod species

Warming intensify CO2 flux and nutrient release from algal wrack subsidies on sandy beaches

Algal wrack subsidies underpin most of the food web structure of exposed sandy beaches and are responsible of important biogeochemical processes that link marine and terrestrial ecosystems. The response in decomposition of algal wrack deposits to global warming has not been studied in ocean-exposed sandy beaches to date. With this aim, passive open top chambers (OTCs) were used to increase soil temperature within the range predicted by the IPCC for western Europe (between 0.5 and 1.5 degrees C), following the hypothesis that the biogeochemical processing of macroalgal wrack subsidies would accelerate in response to temperature increase. The effect of temperature manipulation on three target substrates: fresh and aged macroalgae, and bare sand, was tested. Results indicated that a small warming (<0.5 degrees C) affected the wrack decomposition process through traceable increases in soil respiration through CO2 flux, inorganic nutrients within the interstitial environment (N and P), sediment organic contents measured through the amount of proteins and microbial pool through the total soil DNA. The different responses of soil variables in the studied substrates indicated that the decomposition stage of stranded macroalgae influences the biogeochemical processing of organic matter in sandy beaches. Thus, CO2 fluxes, releases of organic and inorganic nutrients and microbial activity intensify in aged wrack deposits. Our results predict that expected global warming will increase the release of inorganic nutrients to the coastal ocean by 30% for the N (21 Gg/year) and 5.9% for P (14 Gg/year); that increase for the flow of C to the atmosphere as CO2 was estimated in 8.2% (523 Gg/year). This study confirms the key role of sandy beaches in recycling ocean-derived organic matter, highlighting their sensitivity to a changing scenario of global warming that predicts significant increases in temperature over the next few decades.Peer reviewe

Decreased thermal tolerance under recurrent heat stress conditions explains summer mass mortality of the blue mussel Mytilus edulis

Extreme events such as heat waves have increased in frequency and duration over the last decades. Under future climate scenarios, these discrete climatic events are expected to become even more recurrent and severe. Heat waves are particularly important on rocky intertidal shores, one of the most thermally variable and stressful habitats on the planet. Intertidal mussels, such as the blue mussel Mytilus edulis, are ecosystem engineers of global ecological and economic importance, that occasionally suffer mass mortalities. This study investigates the potential causes and consequences of a mass mortality event of M. edulis that occurred along the French coast of the eastern English Channel in summer 2018. We used an integrative, climatological and ecophysiological methodology based on three complementary approaches. We first showed that the observed mass mortality (representing 49 to 59% of the annual commercial value of local recreational and professional fisheries combined) occurred under relatively moderate heat wave conditions. This result indicates that M. edulis body temperature is controlled by non-climatic heat sources instead of climatic heat sources, as previously reported for intertidal gastropods. Using biomimetic loggers (i.e. 'robomussels'), we identified four periods of 5 to 6 consecutive days when M. edulis body temperatures consistently reached more than 30 °C, and occasionally more than 35 °C and even more than 40 °C. We subsequently reproduced these body temperature patterns in the laboratory to infer M. edulis thermal tolerance under conditions of repeated heat stress. We found that thermal tolerance consistently decreased with the number of successive daily exposures. These results are discussed in the context of an era of global change where heat events are expected to increase in intensity and frequency, especially in the eastern English Channel where the low frequency of commercially exploitable mussels already questions both their ecological and commercial sustainability.Funding Agency French Ministere de l'Enseignement Superieur et de la Recherche Region Hauts-de-France European Funds for Regional Economical Development Pierre Hubert Curien PESSOA Felloswhip Fundacao para a Ciencia e Tecnologia (FCT-MEC, Portugal) IF/01413/2014/CP1217/CT0004 National Research Foundation - South Africa 64801 South African Research Chairs Initiative (SARChI) of the Department of Science and Technology National Research Foundation - South Africainfo:eu-repo/semantics/publishedVersio

Relationships between biodiversity and the stability of marine ecosystems: comparisons at a European scale using meta-analysis.

The relationship between biodiversity and stability of marine benthic assemblages was investigated using existing data sets (n = 28) covering various spatial (m-km) and temporal (1973-2006) scales in different benthic habitats (emergent rock, rock pools and sedimentary habitats) through meta-analyses. Assemblage stability was estimated by measuring temporal variances of species richness, total abundance (density or % cover) and community species composition and abundance structure (using multivariate analyses). Positive relationships between temporal variability in species number and richness were generally observed at both quadrat (<1 m2) and site (100 m2) scales, while no relationships were observed by multivariate analyses. Positive relationships were also observed at the scale of site between temporal variability in species number and variability in community structure with evenness estimates. This implies that the relationship between species richness or evenness and species richness variability is slightly positive and depends on the scale of observation, suggesting that biodiversity per se is important for the stability of ecosystems. Changes within community assemblages in terms of structure are, however, generally independent of biodiversity, suggesting no effect of diversity, but the potential impact of individual species, and/or environmental factors. Except for sedimentary and rock pool habitats, no relationship was observed between temporal variation of the aggregated variable of total abundances and diversity at either scale. Overall our results emphasise that relationships depend on scale of measurements, type of habitats and the marine systems (North Atlantic and Mediterranean) considered

The Future of Benthic Indicators : Moving up to the Intertidal

International audienceThe use of benthic indicators has increased dramatically during the last decades. The number of articles published on the subject, as well as the number of citations, has been particularly increasing since the early 90's, notably in relation with the implementation of directives for the management of aquatic/marine ecosystems such as the Water Framework Directive and the Marine Strategy Framework Directive. Current benthic indicators suffer from severe drawbacks and their practical use is still discussed and might have reached a dead end. Indicators based on species composition are not totally satisfactory, mainly because they exhibit a high spatio-temporal variability (e.g. variable at both seasonal and pluri-annual scales) and are user-dependent (e.g. divergent results from US orEuropeexperts.) In turn, modifications of behaviour, metabolism, phenotype or stable isotopes composition in invertebrates usually occur at short time scales, compared to detectable changes in community composition, and makes their use particularly relevant as indicators of perturbation. It is argued in this paper that these functional indicators might be relatively quickly implemented in the intertidal, and represent an effective alternative to current benthic indicators

Pomegranate interest in osteoporosis nutritional prevention : role of lipid and polyphenolic fractions, physiological, cellular and molecular approaches

Dans le contexte actuel d'allongement de l'espérance de vie, la prévalence et le coût de prise en charge des maladies liées à l'âge telles que l'ostéoporose sont de plus en plus importants. Dans le cadre du développement de nouvelles stratégies de prise en charge des pathologies osseuses, la nutrition offre un potentiel évident et représente une excellente solution alternative aux traitements habituels. Bien que trop marginales, les études des activités biologiques de certains aliments ont pourtant d'ores et déjà montré des effets protecteurs sur l'acquisition du capital osseux et sa préservation par le biais de leurs propriétés anti-inflammatoires et antioxydantes. A ce titre, la consommation de grenade est envisagée car elle fait actuellement l'objet d'intérêts scientifiques croissants ayant déjà suggéré ses avantages nutritionnels et pharmacologiques pour la prévention de certaines pathologies chroniques associées au vieillissement. Dans le cadre du projet de thèse, les premiers travaux réalisés, focalisés donc sur la grenade, ont permis de dégager le concept selon lequel les propriétés biologiques de ce fruitexceptionnel seraient expliquées par la composition particulière de ses différentes parties : en polyphénols pour le jus (anthocyanines) et la peau (ellagitannins) et en acide punicique pour les pépins, et surtout au potentiel anti-oxydant et anti-inflammatoire de ces molécules. Cette hypothèse a donné lieu à une première étude à vocation phytochimique qui a permis de déterminer, de standardiser, produire et caractériser deux types d'extraits de grenade titrés en molécules d'intérêt (acide punicique et ellagitannins). Nous avons ensuite étudié l'impact de la consommation du fruit (donné dans sa totalité) ou de ses principales parties (jus et peau) ou encore d'extraits concentrés en ellagitannins et en acide punicique, sur la biologie osseuse, dans un modèle expérimental d'ostéoporose post-ménopausique bien décrit (ostéopénie consécutive à une carence oestrogénique induite par ovariectomie chez des souris C57bl6/j). Cette étude préclinique a permis de montrer que tous les régimessupplémentés en grenade (quelle que soit la partie) permettent de limiter les processus de déminéralisation et l'altération micro-architecturale de l'os, s'expliquant par une réduction des marqueurs de résorption osseuse et une amélioration de ceux de la formation ; et une diminution des paramètres inflammatoires, et oxydants. Ensuite, afin de déterminer les structures médiatrices de ces effets, une approche originale ex-vivo a permis d'étudier plus finement les mécanismes cellulaires ostéoblastiques et ostéoclastiques associés à la consommation des extraits de grenade testés. Cette analyse a été développée de façon à tenir compte du métabolisme particulier de ces micronutriments et donc des modifications systémiques engendrées suite à l'ingestion des extraits. Ce travail a permis de mettre en évidence que les effets observés in-vivo sur l'os,pouvaient être expliqués par une action directe aussi bien sur les ostéoblastes que sur les ostéoclastes, et donc une optimisation du remodelage osseux. De fait, la consommation d'ellagitannins et d'acide punicique augmente l'activité des ostéoblastes (activité alcaline phosphatase et formation de nodules de calcium) et l'expression des principaux acteurs et facteurs de transcription impliqués dans leur différenciation, tout en diminuant l'expression de ceux responsables de la différenciation ostéoclastique. Dans le même temps, l'analyse des transcrits met en évidence que les extraits de grenade favorisent l'activation d'acteurs impliqués dans les mécanismes de défenses contre le stress oxydant et l'inflammation, au niveau du microenvironnement osseux. Ces résultats révèlent pour la première fois le potentiel de la grenade au regard de la physiologie osseuse en proposant ses possibles mécanismes d'actions, via une approche nutritionnelle complète et intégrée respectant la physiologie.In the current context of increased life expectancy, prevalence and socioeconomic consequences of age-related diseases such as osteoporosis represent a major public health problem worldwide. This is why development of new strategies of prevention is highly suitable to provide a wide array of options (alternatives to usual therapies) for health professionals. So far, studies targeting nutrient biological activities have been mainly focused on both calcium and vitamin D. Nevertheless, other nutrients have shown a protective effect on bone mass acquisition and preservation through their anti-inflammatory and antioxidant properties. In this light, pomegranate is endowed with such a potential. Indeed there is an increasing scientific interest that has already suggested its nutritional and pharmacological benefits on prevention of some chronic age-associated diseases. As part of this project, the initial work focused on pomegranate has highlighted the link between its health benefit potential and the exceptional composition of its main parts: polyphenols from the juice (anthocyanins) and the peel (ellagitannins) and punicic acid from seed, those micronutrients being able to elicit antioxidant and anti-inflammatory capacities. Our work hypothesis resulted from a first phytochemical study leading to identify, standardize, characterize and produce two types of pomegranate extracts titrated on 2 molecules of interest (punicic acid and ellagitannins). We thus investigated the outcome of consumption, first of the whole fruit or its principal parts (peel and juice) and in a second time of concentrated ellagitannins and punicic acid extracts on bone biology in a well described experimental model of postmenopausal osteoporosis (osteopenia induced by estrogen deficiency after ovariectomy in C57bl6/j mice). In this preclinical study, wedemonstrated that all the diets supplemented with pomegranate significantly prevented bone loss and micro-architecture impairment. Those findings are associated with transcriptional changes in bone tissue, suggesting involvement of both osteoclastogenesis inhibition and osteoblastogenesis improvement, and reduced inflammatory and oxidative parameters, as well. Then, to determine more accurately the molecules and the signaling pathways involved in those effects, an original ex-vivo study was set up on both osteoblasts and osteoclasts, with respect to physiological conditions (i.e., the aim being to mimic systemic changes and generation of specific circulating metabolites associated with the extractsingestion). This work allowed clarifying the bone sparing effects observed in-vivo. Indeed, pomegranate extracts had the ability to elicit a significant increase in alkaline phosphatase activity, matrix mineralization and transcriptional levels of major osteoblast lineage markersinvolving key signaling pathways, while the expression of specific osteoclast differentiation markers and RANK-RANKL downstream signaling targets were down-regulated. In addition, transcripts analysis revealed that pomegranate extracts were able to induce defense mechanisms against oxidative stress and inflammation in the bone microenvironment. Our results show for the first time the pomegranate potential regarding bone physiology, underlying its possible mechanisms on bone remodeling through a complete and integrated nutritional approach respecting the physiology