In vitro screening of probiotic lactic acid bacteria and prebiotic glucooligosaccharides to select effective synbiotics

Probiotics and prebiotics have been demonstrated to positively modulate the intestinal microflora and could promote host health. Although some studies have been performed on combinations of probiotics and prebiotics, constituting synbiotics, results on the synergistic effects tend to be discordant in the published works. The first aim of our study was to screen some lactic acid bacteria on the basis of probiotic characteristics (resistance to intestinal conditions, inhibition of pathogenic strains). Bifidobacterium was the most resistant genus whereas Lactobacillus farciminis was strongly inhibited. The inhibitory effect on pathogen growth was strain dependent but lactobacilli were the most effective, especially L. farciminis. The second aim of the work was to select glucooligosaccharides for their ability to support the growth of the probiotics tested. We demonstrated the selective fermentability of oligodextran and oligoalternan by probiotic bacteria, especially the bifidobacteria, for shorter degrees of polymerisation and absence of metabolism by pathogenic bacteria. Thus, the observed characteristics confer potential prebiotic properties on these glucooligosaccharides, to be further confirmed in vivo, and suggest some possible applications in synbiotic combinations with the selected probiotics. Furthermore, the distinctive patterns of the different genera suggest a combination of lactobacilli and bifidobacteria with complementary probiotic effects in addition to the prebiotic ones. These associations should be further evaluated for their synbiotic effects through in vitro and in vivo models

A Scalable Genome-Editing-Based Approach for Mapping Multiprotein Complexes in Human Cells

SummaryConventional affinity purification followed by mass spectrometry (AP-MS) analysis is a broadly applicable method used to decipher molecular interaction networks and infer protein function. However, it is sensitive to perturbations induced by ectopically overexpressed target proteins and does not reflect multilevel physiological regulation in response to diverse stimuli. Here, we developed an interface between genome editing and proteomics to isolate native protein complexes produced from their natural genomic contexts. We used CRISPR/Cas9 and TAL effector nucleases (TALENs) to tag endogenous genes and purified several DNA repair and chromatin-modifying holoenzymes to near homogeneity. We uncovered subunits and interactions among well-characterized complexes and report the isolation of MCM8/9, highlighting the efficiency and robustness of the approach. These methods improve and simplify both small- and large-scale explorations of protein interactions as well as the study of biochemical activities and structure-function relationships

A Myrtus communis extract enriched in myrtucummulones and ursolic acid reduces resistance of Propionibacterium acnes biofilms to antibiotics used in acne vulgaris

Recent works present evidence of Propionibacterium acnes growing as a biofilm in cutaneous follicles. This formation of clusters is now considered as an explanation for the in vivo resistance of P. acnes to the main antimicrobials prescribed in acne vulgaris. Purpose : Our objective was to explore this hypothesis and propose a new therapeutic approach focusing on anti-biofilm activity of Myrtacine® New Generation (Mediterranean Myrtle extract–Botanical Expertise P. Fabre) alone or combined with antibiotics. Methods/Results : Using in vitro models able to promote the growth of adhered bacteria, the loss of sensitivity of P. acnes biofilms (48 h) towards erythromycin and clindamycin was checked considering either sensitive or resistant strains. In the same time, the activity of Myrtacine® New Generation against biofilm formation and mature biofilm (48 h) was evaluated. Using a dynamic model of biofilm formation, we noted an inhibition of biofilm formation (addition of Myrtacine® New Generation at T 0) and a significant effect on mature biofilm (48 h) for 5 min of contact. This effect was also checked using the static model of biofilm formation for Myrtacine® New Generation concentrations ranging from 0.03% to 0.0001%. A significant, dose-dependent anti-biofilm effect was observed and notable even at a concentration lower than the active concentration on planktonic cells, i.e. 0.001%. Finally, the interest of the combination of Myrtacine® New Generation with antibiotics was explored. An enhanced efficacy was noted when erythromycin (1000 mg/l) or clindamycin (500 mg/l) was added to 0.001% Myrtacine®, leading to significant differences in comparison to each compound used alone

Proteobacteria from the human skin microbiota: Species-level diversity and hypotheses

The human skin microbiota is quantitatively dominated by Gram-positive bacteria, detected by both culture and metagenomics. However, metagenomics revealed a huge variety of Gram-negative taxa generally considered from environmental origin. For species affiliation of bacteria in skin microbiota, clones of 16S rRNA gene and colonies growing on diverse culture media were analyzed. Species-level identification was achieved for 81% of both clones and colonies. Fifty species distributed in 26 genera were identified by culture, mostly belonging to Actinobacteria and Firmicutes, while 45 species-level operational taxonomic units distributed in 30 genera were detected by sequencing, with a high diversity of Proteobacteria. This mixed approach allowed the detection of 100% of the genera forming the known core skin Gram-negative microbiota and 43% of the known diversity of Gram-negative genera in human skin. The orphan genera represented 50% of the current skin pan-microbiota. Improved culture conditions allowed the isolation of Roseomonas mucosa, Aurantimonas altamirensis and Agrobacterium tumefaciens strains from healthy skin. For proteobacterial species previously described in the environment, we proposed the existence of skin-specific ecotypes, which might play a role in the fine-tuning of skin homeostasis and opportunistic infections but also act as a shuttle between environmental and human microbial communities. Therefore, skin-associated proteobacteria deserve to be considered in the One-Health concept connecting human health to the health of animals and the environment

Seven recommendations to make your invasive alien species data more useful

Science-based strategies to tackle biological invasions depend on recent, accurate, well-documented, standardized and openly accessible information on alien species. Currently and historically, biodiversity data are scattered in numerous disconnected data silos that lack interoperability. The situation is no different for alien species data, and this obstructs efficient retrieval, combination, and use of these kinds of information for research and policy-making. Standardization and interoperability are particularly important as many alien species related research and policy activities require pooling data. We describe seven ways that data on alien species can be made more accessible and useful, based on the results of a European Cooperation in Science and Technology (COST) workshop: (1) Create data management plans; (2) Increase interoperability of information sources; (3) Document data through metadata; (4) Format data using existing standards; (5) Adopt controlled vocabularies; (6) Increase data availability; and (7) Ensure long-term data preservation. We identify four properties specific and integral to alien species data (species status, introduction pathway, degree of establishment, and impact mechanism) that are either missing from existing data standards or lack a recommended controlled vocabulary. Improved access to accurate, real-time and historical data will repay the long-term investment in data management infrastructure, by providing more accurate, timely and realistic assessments and analyses. If we improve core biodiversity data standards by developing their relevance to alien species, it will allow the automation of common activities regarding data processing in support of environmental policy. Furthermore, we call for considerable effort to maintain, update, standardize, archive, and aggregate datasets, to ensure proper valorization of alien species data and information before they become obsolete or lost

The Genome Sequence of the Grape Phylloxera Provides Insights into the Evolution, Adaptation, and Invasion Routes of an Iconic Pest

Background: Although native to North America, the invasion of the aphid-like grape phylloxera Daktulosphaira vitifoliae across the globe altered the course of grape cultivation. For the past 150 years, viticulture relied on grafting-resistant North American Vitis species as rootstocks, thereby limiting genetic stocks tolerant to other stressors such as pathogens and climate change. Limited understanding of the insect genetics resulted in successive outbreaks across the globe when rootstocks failed. Here we report the 294-Mb genome of D. vitifoliae as a basic tool to understand host plant manipulation, nutritional endosymbiosis, and enhance global viticulture. Results: Using a combination of genome, RNA, and population resequencing, we found grape phylloxera showed high duplication rates since its common ancestor with aphids, but similarity in most metabolic genes, despite lacking obligate nutritional symbioses and feeding from parenchyma. Similarly, no enrichment occurred in development genes in relation to viviparity. However, phylloxera evolved > 2700 unique genes that resemble putative effectors and are active during feeding. Population sequencing revealed the global invasion began from the upper Mississippi River in North America, spread to Europe and from there to the rest of the world. Conclusions: The grape phylloxera genome reveals genetic architecture relative to the evolution of nutritional endosymbiosis, viviparity, and herbivory. The extraordinary expansion in effector genes also suggests novel adaptations to plant feeding and how insects induce complex plant phenotypes, for instance galls. Finally, our understanding of the origin of this invasive species and its genome provide genetics resources to alleviate rootstock bottlenecks restricting the advancement of viticulture

Interrelation entre le complexe MRE11-RAD50-NBS1 et FANCD2, une protéine de l'anémie de Fanconi

L'anémie de Fanconi est une maladie génétique récessive rare, caractérisée par une défaillance de la moelle osseuse, des anomalies du développement et une forte incidence de cancer, essentiellement des leucémies myéloïdes aiguës. L'anémie de Fanconi peut être causée par la mutation de 13 gènes différents, et neuf des protéines Fane sont responsables de la monoubiquitination de FANCD2, qui est donc centrale dans la 'voie Fanconi'. Le complexe MRN joue plusieurs rôles essentiels dans la réparation des cassures double-brin dans l'ADN par recombinaison homologue : dans la signalisation du dommage et dans les points de contrôle du cycle cellulaire, dans la préparation des extrémités de la cassure pour les étapes suivantes de la réparation ainsi qu'un rôle structural afin de faciliter la réparation. Il a été montré précédemment que la protéine FANCD2 colocalise avec NBS1. Cependant, la relation fonctionnelle entre FANCD2 et MRE11 est mal comprise. Mes travaux de doctorat montrent que l'inhibition de MRE11, NBS1 ou RAD50 conduit à la déstabilisation de FANCD2. FANCD2 s'accumule de la phase S à la phase G2 aux sites contenant de l'ADN simple brin ou aux sites de dommages à l'ADN. La protéine FANCD2 purifiée lie préférentiellement d'ADN simple brin en comparaison à différents substrats d'ADN. L'inhibition de l'activité nuclease de MRE11 par le MIRIN diminue le nombre de foyers de FANCD2 formés in vivo. Nous proposons donc que FANCD2 lie l'ADN simple brin provenant de la résection de la cassure double brin par MRE11. Nos données établissent que MRN est un régulateur essentiel de la stabilité et de la fonction de FANCD2 au cours de la réponse aux dommages à l'ADN

Impacts des cyanobactéries d'eau douce dans la santé publique

Les cyanobactéries aussi appelées algues bleu-vert, algues d'eau douce, cyanophycées sont de plus en plus étudiées dans le monde entier. Pour mieux comprendre leur développement dans les eaux de surface, mais aussi dans les réservoirs d'eau, il est nécessaire de connaître leur biologie. L'intérêt de l'étude de ces cyanobactéries repose sur le fait qu'elles sont toxiques pour l'homme : toxicité hépatique, gastro-intestinale, dermique, neurologique. De plus en plus fréquentes à cause, entre autre, de la pollution, de l'eutrophisation des eaux, les cyanobactéries deviennent un problème de santé publique, qui amène différents pays à prendre des résolutions juridiques ou non. En France cette toxicité est méconnue des professionnels de santé, d'où une inexistence de données épidémiologiques. Il est donc important d'envisager d'informer les médecins, les pharmaciens, afin de les sensibiliser au problème, d'où la proposition d'une plaquette informative.TOULOUSE3-BU Santé-Centrale (315552105) / SudocTOULOUSE3-BU Santé-Allées (315552109) / SudocSudocFranceF