Wnt and Neuregulin1/ErbB signalling extends 3D culture of hormone responsive mammary organoids

The development of in vitro culture systems quantitatively and qualitatively recapitulating normal breast biology is key to the understanding of mammary gland biology. Current three-dimensional mammary culture systems have not demonstrated concurrent proliferation and functional differentiation ex vivo in any system for longer than 2 weeks. Here, we identify conditions including Neuregulin1 and R-spondin 1, allowing maintenance and expansion of mammary organoids for 2.5 months in culture. The organoids comprise distinct basal and luminal compartments complete with functional steroid receptors and stem/progenitor cells able to reconstitute a complete mammary gland in vivo. Alternative conditions are also described that promote enrichment of basal cells organized into multiple layers surrounding a keratinous core, reminiscent of structures observed in MMTV-Wnt1 tumours. These conditions comprise a unique tool that should further understanding of normal mammary gland development, the molecular mechanism of hormone action and signalling events whose deregulation leads to breast tumourigenesis

Modeling colorectal cancer: A bio-resource of 50 patient-derived organoid lines

Background and Aim Colorectal cancer (CRC) is the second leading cause of cancer death worldwide. To improve outcomes for these patients, we need to develop new treatment strategies. Personalized cancer medicine, where patients are treated based on the characteristics of their own tumor, has gained significant interest for its promise to improve outcomes and reduce unnecessary side effects. The purpose of this study was to examine the potential utility of patient-derived colorectal cancer organoids (PDCOs) in a personalized cancer medicine setting. Methods Patient-derived colorectal cancer organoids were derived from tissue obtained from treatment-naïve patients undergoing surgical resection for the treatment of CRC. We examined the recapitulation of key histopathological, molecular, and phenotypic characteristics of the primary tumor. Results We created a bio-resource of PDCOs from primary and metastatic CRCs. Key histopathological features were retained in PDCOs when compared with the primary tumor. Additionally, a cohort of 12 PDCOs, and their corresponding primary tumors and normal sample, were characterized through whole exome sequencing and somatic variant calling. These PDCOs exhibited a high level of concordance in key driver mutations when compared with the primary tumor. Conclusions Patient-derived colorectal cancer organoids recapitulate characteristics of the tissue from which they are derived and are a powerful tool for cancer research. Further research will determine their utility for predicting patient outcomes in a personalized cancer medicine setting

Effect of catch crop and animal manure amendment on the carbon sequestration, nitrates and dissolved organic carbon losses in subsoil

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Stromal Niche Signals That Orchestrate Intestinal RegenerationSummary

Stromal cell populations have a central role in providing signals that support the maintenance, differentiation, and function of the intestinal epithelium. The behavior and fate of epithelial cells is directed by the spatial organization of stromal cells that either sustain stem and progenitor cell identity or drive differentiation. A combination of single-cell analyses, mouse models, and organoid coculture assays have provided insight into the diversity of signals delivered by stromal cells. Signaling gradients are established and fine-tuned by the expression of signaling agonists and antagonists along the crypt-villus axis. On epithelial injury, there are disruptions to the abundance and organization of stromal populations. There are also distinct changes in the signals originating from these cells that impact remodeling of the epithelium. How these signals coordinate to mediate epithelial repair or sustain tissue injury in inflammatory bowel diseases is beginning to emerge. Understanding of these processes may lead to opportunities to target stromal cell populations as a strategy to modify disease states

Prioritisation of veterinary pharmaceuticals prior to a monitoring campaign into water resources and drinking water: Case of Brittany, an intensive husbandry area

International audiencePharmaceuticals residues are nowadays of growing concern, especially in aquatic environments. Numerous and detailed researches have been conducted on human pharmaceutical residues at national and european scale, while ocurrence of veterinary pharmaceutical residues remains still largely unknown. Brittany is a region subjected to high animal husbandry pressures. Veterinary pharmaceuticals residues can enter the environment directly or indirectly during pastures or spreading of animal manure on soils. Thus, aquatic environments in Brittany are sensitive spots, potentially contaminated by a lot of veterinary residues, such as antibiotics or antiparasitic drugs. The project aims at realizing an overview of the contamination (types molecules and levels of concentrations) of raw water resources subjected to a strong agricultural pressure, and into drinking water obtained from those resources. In order to realize a monitoring study during one year, the first step was to perform a prioritization of veterinary pharmaceuticals, to select the veterinary pharmaceuticals which are the most susceptible to reach and to be detected in the aquatic environment

New Insights into Anticarcinogenic Properties of Adiponectin: A Potential Therapeutic Approach in Breast Cancer?

Document Type : Book ChapterInternational audienceObesity is a recognized breast cancer risk factor in postmenopausal women. A recent hypothesis suggests a major role for adipose tissue in carcinogenesis. During many years, the adipose tissue was only considered as a fat storage of energy. This tissue is now described as an endocrine organ secreting a large range of molecules called adipokines. Among these adipokines, adiponectin may play a major role in breast cancer. Plasma adiponectin levels were found to be decreased in cases of breast cancer and in obese patients. Adiponectin may act directly on breast cancer cells by inhibiting proliferation and angiogenesis or by stimulating apoptosis. Increasing adiponectin levels may be of major importance in the prevention and/or the treatment of breast cancer. This therapeutic approach may be of particular significance for obese patients. The beneficial effects of adiponectin and its possible therapeutic applications will be discussed in this review

Résidus de médicaments vétérinaires dans les eaux destinées à la consommation humaine en zones d’élevage intensif: Cas des bassins versants bretons

National audienceContexteLa présence de résidus de médicaments dans les ressources en eau participe pour partie à l’exposition de la population (Mompelat et al., 2009) et représente une menace diffuse et potentiellement persistante dans les hydrosystèmes. Des études ont été menées sur les résidus médicamenteux à usage humain au niveau national (campagnes de l'ANSES, des Agences de l’Eau, du BRGM de l’IRSTEA etc. ; thèses de A. Togola (2006), de F. Tamtam (2008) et de S. Mompelat (2011)) ou encore au niveau européen (PHARMAS 2011-2014, Cytothreat 2011-2014), mais peu d’études se sont intéressées aux résidus médicamenteux vétérinaires (RMV).En France, la Bretagne est une région d'élevage intensif qui possède un chevelu hydrographique très dense et vulnérable, et 75% des eaux destinées à la consommation humaine (EDCH) sont issues d’eau de surface (ARS Bretagne, 2017). Le recyclage important des matières organiques exogènes sur les sols pour valoriser l’azote, le phosphore et le carbone peut générer une pollution diffuse car ces effluents peuvent contenir des quantités importantes de résidus de médicaments. Les RMV peuvent être transférés vers ces ressources en eau par lixiviation ou ruissellement à partir de parcelles pâturées, ou ayant reçu des effluents d’élevage mais également par le biais d’activités piscicoles.L’objectif du projet EXPOVETO (2016-2018) est d’identifier et quantifier les RMV présents dans les ressources en EDCH et les eaux traitées correspondantes en Bretagne.MéthodeMolécules d’intérêt : 37 RMV hiérarchisés à partir de 4 critères : avec les pratiques vétérinaires bretonnes (Soulier et al., 2015, ARS/DREAL 2013-2015), les données sur leur pharmacocinétique et leur devenir dans l’environnement et la faisabilité analytique (Thèse de Lise Charuaud, 2016-2018).Sites sélectionnés : 25 captages EDCH en Bretagne (23 eaux de surface et 2 eaux souterraines), situés en contexte d’élevage intensif ont été retenus (ARS/DREAL 2013-2015).Échantillonnage : La stratégie d’échantillonnage a pour objectif de refléter les variations des prescriptions vétérinaires, les périodes d’épandages et le régime hydrique (étiage ou hautes eaux) (Jaffrézic et al., 2017). Quatre campagnes de prélèvements (mars 2017 ; mai 2017 ; septembre 2017 et janvier 2018), en entrée et sortie d’usine de potabilisation (200 échantillons) ont été réalisées sur les sites d’intérêts.Analyses : Les RMV sont extraits des échantillons d’eau sur support solide suivie d’une séparation par chromatographie liquide couplée à une détection par spectrométrie de masse en tandem.RésultatsPour les 3 premières campagnes, des RMV ont été quantifiés dans les ressources en eau (23 eaux de surfaces et 2 eaux souterraines) dans 32% à 47% des échantillons selon les campagnes. La diversité de molécules quantifiées est plus importante en période d’étiage (10 molécules).Les concentrations mesurées varient de 5 ng/l (la limite de quantification) à 2 μg/l.Dans les eaux traitées correspondantes, des RMV sont également quantifiés de 4% jusqu’à 65% des échantillons selon les campagnes. La diversité des molécules quantifiées était plus importante en période d’épandages. Les concentrations mesurées varient de 5ng/l à 200 ng/l environ. La 4eme campagne (hautes eaux) est en cours d’analyse, les résultats seront comparés avec les 3 premières campagnes

Effects of biogas digestate and winter crops on dissolved organic carbon and nitrates fluxes in soil.

International audienceThe anaerobic digestion industry is currently in development and enables the energetic valorization of organic waste products (OWP) to generate biogas. A co-product of this process, digestate, is more and more produced and can be recycled on agricultural lands as a low-cost alternative to mineral fertilizers. Thus, the organic carbon (C) it contains could improve the soil quality and store C in subsoil through dissolved organic C (DOC) transfers. Biogas digestate is a recent OWP characterized by a chemical composition rich in recalcitrant molecules and little is known about its impacts on dissolved fluxes in the scientific literature.In order to evaluate the impacts of biogas digestate and winter crop on DOC and nitrates fluxes in soil, the experimental site EFELE from the SOERE PRO network (https://www6.inra.fr/valor-pro) was followed. The long-term evolution of the repeated application of OWP since 2012 was assessed for three different treatments: pig slurry, biogas digestate from the anaerobic digestion of pig slurry and a mineral fertilizer as a control. Lysimeters were monitored from 2014 to 2022 (8 drainage seasons) with two replicates per modality and depth (40 and 90 cm) under winter cover of wheat or mustard. The drainage seasons lasted from November to April. The DOC and nitrates dynamics observed for the four years under wheat and the four years under mustard were repeatable. Nitrates leaching losses were low under mustard (2.7 ± 2.3 kg N.ha-1 at 40cm and 0.9 ± 0.8 at 90cm), which had fulfilled its role as a catch crop, and higher under wheat (30.5 ± 12.9 kg N.ha-1 at 40cm and 14.3 ± 10.1 at 90cm), regardless of treatment. Regarding the DOC fluxes at 40cm depth, there was a significant difference between mustard (34.0 ± 11.4 kg C.ha-1) and wheat (20.6 ± 11.1 kg C.ha-1). DOC fluxes under biogas digestate were significantly higher (36.3 ± 16.2 kg C.ha-1) than under mineral fertilizer and pig slurry (21.4 ± 7.4 kg C.ha-1 on average). At the beginning of the drainage season, the DOC concentrations dynamics were already significantly different between digestate and the two other treatments under mustard, which was sown in September and already well developed. Under wheat, which was sown in November, there was no difference between treatments at the beginning of the season. It was only in the middle of winter, when the crop had more time to develop, that a difference could be observed. It seemed that the crop effect controlled the treatment effect. DOC fluxes to groundwater (90 cm) were not significantly different between treatments or crop (7.4 ± 6.0 kg C.ha-1 on average). Part of the C could be stored between 40cm and 90cm depth or mineralized

Veterinary pharmaceutical residues in water resources and tap water in an intensive husbandry area in France

International audienceIn intensive livestock areas, veterinary pharmaceutical residues (VPRs) can occur in water resources, but also in tap water because treatment processes are not designed to remove these contaminants. The main objective of this study is to assess the occurrence of VPRs in water resources and tap waters in Brittany. As several identical compounds are used in both veterinary and human medicine, a toolbox (stanols and pharmaceuticals) is used to help determine the origin of contamination in the case of mixed-use molecules. Water resources samples were collected from 25 sites (23 surface waters and two groundwaters) used for tap water production and located in watersheds considered as sensitive due to intensive husbandry activities. Samples were also taken at 23 corresponding tap water sites. A list of 38 VPRs of interest was analyzed. In water resources, at least one VPR was quantified in 32% of the samples. 17 different VPRs were quantified, including antibiotics, antiparasitic drugs and anti-inflammatory drugs. Concentration levels ranged between 5 ng/L and 2946 ng/L. Mixed-use pharmaceuticals were quantified in twelve samples of water resources and among these samples nine had a mixed overall fecal contamination. In the context of this large-scale study, it appeared difficult to determine precisely the factors impacting the occurrence of VPRs. VPRs were quantified in 20% of the tap water samples. Twelve VPRs were quantified, including ten compounds exclusively used in veterinary medicine and two mixed-use compounds. Concentration levels are inferior to 40 ng/L for all compounds, with the exception of the antibiotic florfenicol which was quantified at 159 ng/L and 211 ng/L. The population of Brittany may therefore be exposed to these contaminants through tap water. These observations should be put into perspective with the detection frequencies per compound which are all below 10% in both water resources and tap water

Adaptations of hepatic amino acid uptake and net utilisation contributes to nitrogen economy or waste in lambs fed nitrogen- or energy-deficient diets

International audienceWe investigated the effect of relative changes in dietary nitrogen (N) and energy supply and the subsequent variations in net portal appearance (NPA) of nitrogenous and energy nutrients on the net amino acid (AA) uptake by the liver and net N supply to the peripheral tissues. Six lambs were catheterised across the splanchnic tissues and received, in a replicated Latin square, one of three dietary treatments. The diets were formulated to either match the requirements of N and energy (C), or supply only 0.8 of the N requirement (LN) or 0.8 of the energy requirement (LE). Net fluxes of AA and urea-N were measured across the portal-drained viscera, and estimation of arterial hepatic flow allowed the estimation of hepatic fluxes. Catheters were implanted into the portal and hepatic veins as well as in the abdominal aorta for the measurement of AA fluxes. Animals fed the LN diet showed more efficient N retention (0.59 of digested N) than did the C and LE diet (0.50 and 0.33, respectively; P < 0.001). The NPA of total AA-N for the LN diet was only 0.60 of the value measured for the control (C) diet (P < 0.01). Despite this, the total estimated AA-N net splanchnic fluxes were not significantly different across the three diets (3.3, 1.9 and 2.6 g total AA-N/day for C, LN and LE, respectively, P = 0.52). Thus, different metabolic regulations must have taken place across the liver between the three experimental diets. A combination of decreased net uptake of total AA-N by the liver of animals in the LN diet (0.61 of the C diet; P = 0.002) and reduced urinary urea-N production (0.52 of the C diet; P = 0.001) spared AA from catabolism in the LN diet relative to the other two diets. For the LE diet, the urinary urea-N output was 1.3 times the value of the C diet (P = 0.01). This may relate to an increased catabolism of AA by the muscle and/or, to a lesser extent, to an increased utilisation of AA for gluconeogenesis in the liver. These effects may explain the reduced whole body protein retention observed with the LE diet