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•Plastic particles crossing the intestinal barrier is debated.•PS-NP particles were tested in ex-vivo time series experiments using Ussing chambers.•Two PS-NP concentrations were ...tested on two locations of adult Seabass guts.•PS-NP directly crossed the intestinal barrier of adult Seabass within few minutes.•PS-NP translocation was confirmed by 3 complementary techniques.
Plastic pollution in marine ecosystems constitutes an important threat to marine life. For vertebrates, macro/microplastics can obstruct and/or transit into the airways and digestive tract whereas nanoplastics (NPs; < 1000 nm) have been observed in non-digestive tissues such as the liver and brain. Whether NPs cross the intestinal epithelium to gain access to the blood and internal organs remains controversial, however. Here, we show directly NP translocation across the intestinal barrier of a fish, the European seabass, Dicentrarchus labrax, ex vivo. The luminal side of median and distal segments of intestine were exposed to fluorescent polystyrene NPs (PS-NPs) of 50 nm diameter. PS-NPs that translocated to the serosal side were then detected quantitatively by fluorimetry, and qualitatively by scanning electron microscopy (SEM) and pyrolysis coupled to gas chromatography and high-resolution mass spectrometry (Py-GC-HRMS). Fluorescence intensity on the serosal side increased 15–90 min after PS-NP addition into the luminal side, suggesting that PS-NPs crossed the intestinal barrier; this was confirmed by both SEM and Py-GC-HRMS. This study thus evidenced conclusively that NPs beads translocate across the intestinal epithelium in this marine vertebrate.
Non‐technical summary In this study, we demonstrated that supplementation of the maternal diet with a particular fatty acid, 18:3 n‐3, the precursor of the n‐3 fatty acid family, modified intestinal ...permeability, probably via diet‐induced neuroplastic changes of the enteric nervous system of newborn piglets. These findings suggest that feeding fatty acids of the n‐3 family during pregnancy and lactation impact newborn intestinal barrier function. However, the beneficial versus harmful consequences of this increased intestinal permeability remain to be elucidated.
The intestinal epithelial barrier (IEB) plays a key role in the maintenance of gut homeostasis and the development of the immune system in newborns. The enteric nervous system (ENS), a key regulator of gastrointestinal functions, has been shown to be modulated by nutritional factors. However, it remains currently unknown whether maternal diet, in particular n–3 polyunsaturated fatty acids (n–3PUFAs), can impact upon the IEB in newborn piglets and whether the ENS is involved in this effect. Sows received either a control diet (lard based) or an n–3PUFA diet (linseed oil based) during gestation and lactation. Intestinal paracellular permeability was assessed in Ussing chambers on piglets at birth, 3, 7, 14, 21 and 28 postnatal days (PND). Basal jejunal permeability increased significantly and similarly in both groups until PND14 and decreased thereafter. However, at PND28, permeability was higher in n–3PUFA animals as compared to controls. In addition, a vasoactive intestinal peptide (VIP) receptor antagonist increased paracellular permeability in controls but not in n–3PUFA piglets. Conversely, atropine and hexamethonium decreased paracellular permeability in the n–3PUFA group but not in the control group. Moreover, the n–3PUFA diet increased the proportion of choline acetyltransferase (ChAT)‐immunoreactive (IR) neurons and decreased the proportion of VIP‐IR neurons in the submucosal plexus of piglet jejunum compared to controls. In addition, in primary culture of rat ENS, we showed that 20:5n–3 but not 18:3n–3 increased the proportion of ChAT‐IR neurons and decreased the proportion of VIP‐IR neurons. In conclusion, supplementation of the maternal diet with n–3PUFAs modified intestinal permeability probably via diet‐induced neuroplastic changes in the ENS of newborn piglets.
n-3 polyunsaturated fatty acids (n-3 PUFA) contribute to the normal growth and development of numerous organs in the piglet. The fatty acid composition of piglet tissues is linked to the fatty acid ...composition of sow milk and, consequently, to the composition of sow diet during the gestation and lactation period. In this study, we investigated the impact of different contents of extruded linseed in the sow diet on the fatty acid composition and desaturase gene expression of piglets. Sows received a diet containing either sunflower oil (low 18:3n-3 with 18:3n-3 representing 3% of total fatty acids) or a mixture of extruded linseed and sunflower oil (medium 18:3n-3 with 9% of 18:3n-3) or extruded linseed (high 18:3n-3 with 27% of 18:3n-3) during gestation and lactation. Fatty acid composition was evaluated on sow milk and on different piglet tissues at days 0, 7, 14, 21 and 28. The postnatal evolution of delta5 (D5D) and delta6 (D6D) desaturase mRNA expression was also measured in the liver of low 18:3n-3 and high 18:3n-3 piglets. The milk of high 18:3n-3 sows had higher proportions of n-3PUFA than that of low 18:3n-3 and medium 18:3n-3 sows. Piglets suckling the high 18:3n-3 sows had greater proportions of 18:3n-3, 20:5n-3, 22:5n-3 and 22:6n-3 in the liver, and of 22:5n-3 and 22:6n-3 in the brain than low 18:3n-3 and medium 18:3n-3 piglets. D5D and D6D mRNA expressions in piglet liver were not affected by the maternal diet at any age. In conclusion, extruded linseed in the sow diet modifies the n-3PUFA status of piglets during the postnatal period. However, a minimal content of 18:3n-3 in the sow diet is necessary to increase the n-3PUFA level in piglet liver and brain. Moreover, modifications in the n-3PUFA fatty acid composition of piglet tissue seem linked to the availability of 18:3n-3 in maternal milk and not to desaturase enzyme expression.
Thymol and cinnamaldehyde, extracted from thyme and cinnamon respectively, have multiple effects on mammalian cells. Although the intestinal mucosa is one of the first tissues they are in contact ...with when ingested, their effect on intestinal epithelial cells and especially ion secretion has not been established yet. The aim of this study was to investigate the effect of those two substances on electrolyte secretion and absorption across the porcine jejunal epithelium in Ussing chambers. Jejunal tissues from piglets were mounted in Ussing chambers and the short circuit current measured (I(sc)) after addition of thymol or cinnamaldehyde. Thymol and cinnamaldehyde induced a dose-dependent increase is I(sc). The effect of thymol was inhibited in low Cl-, HCO3(-) free or low Cl-/ HCO3(-) free buffers. It was completely blocked when tissues were previously incubated with tetrodotxin and partially inhibited with hexamethonium. Cinnamaldehyde effect was inhibited when HCO3(-) free or low Cl-/ HCO3(-) free buffers were used. It was not affected by tetrodotoxin but reduced by hexamethonium, suggesting direct activation of receptors on epithelial cells. In conclusion, thymol induces Cl- and HCO3(-) secretion via activation of nervous nicotinic receptors while cinnamaldehyde induces HCO3(-) secretion probably via direct activation of nicotinic receptors on epithelial cells.
In 2015, Annecy Hospital was the first French hospital to perform non-heartbeating organ donation from a Maastricht category III donor (patient awaiting cardiac arrest after withdrawal of treatment). ...Non-heartbeating organ donation (NHBD), performed in France since 2006, had initially excluded this category, due to ethical questions concerning end of life and treatment withdrawal, as well as technical specificities linked to this procedure. Grenoble University Hospital and Edouard-Herriot Hospital in Lyon then performed the first kidney transplants, with satisfactory outcomes in both recipients. This article presents the details and results of this new experience, challenging both on a deontological and organizational level. Functional outcomes of kidney grafts from NHBD are now well known in the literature and confirm their benefit for patients, with similar results to those from heartbeating donors (HBD). International experiences concerning specifically Maastricht category III NHBD are encouraging and promising.
The metabolism and distribution of 2,4-dichloroaniline (2,4-DCA), prochloraz and 4-n-nonylphenol diethoxylate (NP2EO) were investigated in vivo and in vitro in rainbow trout (
Oncorhynchus mykiss). ...Each compound was administered p.o. (10 mg/kg wet weight) and urine was collected during 48 h (2,4-DCA, prochloraz) or 72 h (NP2EO). Fish were sacrificed, the gall bladder was excised and radioactivity was measured in tissues, viscera and carcasses. Metabolic profiles were performed by radio-HPLC and when possible metabolites were identified by LC/MS. For comparison, the biotransformation of these xenobiotics was also investigated in freshly isolated hepatocytes. The metabolic pathways of 2,4-DCA have been identified leading to the glucuronide conjugate (in vivo) and to the glucuronide conjugate and the hydroxylamine metabolite (in vitro). This difference highlights the usefulness of the hepatocyte system in metabolic studies, since the formation of the hydroxylamine reactive metabolite cannot be demonstrated in vivo. For prochloraz, we observed that residue levels are significantly higher in males than in females for gill, fat, brain and carcasses, however, the reasons for this difference remain unclear. Although, the presence of glucuronide conjugates was detected in vivo and in vitro, the chemical structure of isolated metabolites has to be determined. However, the comparison of the in vivo versus in vitro metabolic profiles indicates that several peaks, probably corresponding to intermediate metabolites, were present only in hepatocyte incubations. Biotransformation of NP2EO occurred in vivo and in vitro in rainbow trout, but did not result in the formation of 4-n-NP. The major metabolite present in bile corresponded to the NP2EO-glucuronide but this metabolite was not found in vitro. It is concluded that hepatocytes may produce a different metabolic pattern than in the whole fish, but may also give evidence of a metabolic pathway difficult to apprehend in vivo.
L’obésité et le surpoids sont associés à une inflammation de bas-grade en lien avec des perturbations de la barrière intestinale et du microbiote. Le maintien de l’homéostasie intestinale est soutenu ...par la fonction mitochondriale des cellules épithéliales intestinales (CEI), puisqu’elle fournit l’énergie nécessaire à l’absorption des nutriments, au renouvellement épithélial et au maintien de la barrière intestinale. Or, la consommation d’un régime obésogène est susceptible d’altérer la fonction mitochondriale, tel que cela a été montré au niveau hépatique. Le but de cette étude est d’évaluer si la consommation d’un régime obésogène altère la fonction mitochondriale des CEI de jéjunum chez la souris.
Des souris mâles C57Bl/6J ont été nourries pendant 22 semaines avec un régime contrôle pauvre en lipides (CTRL, lipides=10 % de l’énergie, c=9) ou un régime obésogène (WD, lipides=58 % de l’énergie, et eau de boisson supplémentée en fructose et saccharose (42g/L), CO039. =8)). À l’issue du régime, le jéjunum a été prélevé et les CEI, sous forme de cryptes et de villosités, ont été isolées pour analyser la fonction mitochondriale par la technologie Seahorse® et expressions géniques et protéiques.
Les souris WD ont développé une obésité caractérisée par une augmentation de 50 % du poids corporel (CTRL : 32,5±0,9 vs WD : 46,4±1,8g, p<0,0001) et de l’index d’adiposité viscérale (2,2±0,3 vs 6,7±0,4 %, p<0,0001), une stéatose hépatique et une augmentation de l’endotoxémie (LPS-BP : 535,8±38,5 vs 668,7±39,7ng/mL, p<0,05). L’analyse de la fonction mitochondriale des CEI des souris WD a révélé une diminution de moitié de la respiration basale (60,6±8,9 vs 33,3±6,04pmol d’O2/min/ADN, p<0,05) ainsi qu’une augmentation de la capacité respiratoire de réserve (CRC, 4,01±3,1 vs 31,7±4,5pmol d’O2/min/ADN, p<0,01) associée à une meilleure efficacité de couplage (40,1±2,2 % vs 63,7±6,6 %, p<0,01). En revanche, les CEI des souris WD présentaient une diminution significative des expressions géniques et protéiques de certaines sous-unités des complexes de la chaîne respiratoire mitochondriale. De plus, Pgc1-α, régulateur majeur de la biogénèse mitochondriale, était 60 % moins exprimé chez les souris WD par rapport aux CTRL (P<0,001) tandis que l’expression des gènes impliqués dans la fusion et la fission mitochondriale (Mfn2, Opa1, Fis1 et Pink1) est également diminuée de 10 à 50 %. Enfin, des comptages en microscopie électronique confirment une diminution par deux du nombre de mitochondries par entérocyte de souris WD (51±15 vs 25±16, p<0,0001).
Ainsi, la consommation d’un régime obésogène chez la souris induit l’augmentation de la CRC des CEI de jéjunum et de l’efficacité de couplage, signes d’une meilleure adaptation face à un stress métabolique. Néanmoins, ces cellules sont marquées par une diminution du nombre de mitochondries, de l’expression des complexes mitochondriaux et par une altération de la dynamique mitochondriale, tout ceci concourant à la nette diminution de respiration basale des CEI. Ces altérations pourraient contribuer à la perte de l’homéostasie intestinale et à l’inflammation de bas-grade puis systémique à l’origine des troubles métaboliques chez l’obèse.
The metabolic pattern of the imidazole fungicide prochloraz N-propyl-N-2-(2,4,6-trichlorophenoxy)ethylimidazole-1-carboximide was investigated in rainbow trout (Oncorhynchus mykiss). Following a ...single oral administration of 14Cprochloraz, levels 4.3 ± 4.1 and 3.9 ± 1.8% of the dose were excreted in the bile after 48 h in male and female animals, respectively. Urinary radioactivity accounted for 1.3 ± 0.4 and 2.4 ± 1.1% of the dose over the same period in males and females. Metabolites from both matrices were separated by reversed-phase HPLC with radioactive detection and analyzed by positive and/or negative electrospray ionization mass spectrometry. No unchanged prochloraz was detected in the analyzed excreta. The major biotransformation products in bile were the aldehyde corresponding to the cleavage of the imidazole ring, N-2-(2,4,6-trichlorophenoxy)ethylurea, and the glucuronide conjugate of 2,4,6-trichlorophenoxyethanol. In urine, the major metabolite was 2,4,6-trichlorophenoxyacetic acid. On the basis of enzymatic hydrolysis by β-glucuronidase and LC-MS analyses, this study demonstrates that rainbow trout are able to biotransform prochloraz, mainly as glucuronide conjugates. Keywords: Metabolism; prochloraz; trout; ESI-MS/MS
This study investigated intestinal physiology of piglets at weaning. Piglets (n = 60) weaned at 21 d were food deprived for 2 d and then tube-fed using 2 different diets (a conventional diet vs. a ...wheat-enriched diet). They were slaughtered at d 0, 2, 5, 8, or 15 postweaning. Jejunum, ileum, and colon were mounted in Ussing chambers. In addition, segments of the proximal jejunum of 4 growing pigs were studied 35 d after weaning. Secretory function was assessed by basal short-circuit current (Isc) and secretagogue-stimulated Isc. Glucose absorption was measured by the increase in Isc after the addition of glucose. Epithelial barrier function was measured by transmucosal resistance (R) and horseradish peroxidase (HRP) fluxes across the epithelium. There were no significant differences between the pigs fed the 2 diets for any of the parameters studied. As already reported, a transient villous atrophy was observed. At the same time, we observed an increased basal Isc in jejunum and colon, increased glucose absorption and a dramatic drop of R in jejunum. These parameters had returned to preweaning values by d 5. Weaning was also followed by long-lasting modifications. In jejunum, responses to the secretagogues and glucose absorption were decreased at wk 2 after weaning and were not different between d 15 and 35. Ileal transmucosal resistance increased on d 5 and was stable thereafter. HRP flux in jejunum declined on d 2 and stayed at this low level throughout the experiment. We conclude that weaning induces transient dramatic changes in intestinal physiology but is also a period of maturation of the intestine. J. Nutr. 134: 2256–2262, 2004.
To evaluate the diagnostic performance and the benefit in terms of management of low-dose CT for the imaging assessment of renal colic (CN) emergencies.
Two hundred and ninety-one patients admitted ...to emergency for CN were included in this study. Eighty-seven had a low-dose CT and 40 an ASP and an ultrasound (ASPE). Different parameters evaluating the diagnostic performance and efficiency of care were compared between the two groups. The quantitative and qualitative variables were compared by Student t test and χ(2) test, respectively.
CT and ASPE confirmed the diagnosis of CN in 76% and 54% of patients, respectively (p=0.013). The average lengths of stay were 408 minutes versus 520 (p=0.013) in group scanner and ASPE, respectively. The scan was obtained more rapidly (139 min versus 224, p=0.002). There were more requests for expert advice (30% versus 20%, p=0.18) and gestures endo-urology (9.5% versus 5%, p=0.31) in the CT group compared to the group ASPE. Finally, the patients in the scanner have less painful recurrences (6% versus 12.5%, p=0.18) and fewer imaging examinations of second-line (0% versus 30%, p<0.001).
The low dose CT has been more efficient than the couple ASPE for a CN diagnosis. It optimizes the management of emergency patients by reducing their length of stay, waiting time and the rate of second consultation.
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