Intestinal apoptotic changes linked to metabolic status in fasted and refed rats

Intestinal apoptosis and expression of apoptosis inducers - the cytokines TNFalpha, TGFbeta1 - and the intestinal transcription factor Cdx2, were studied according to two different metabolic and hormonal phases which characterize long-term fasting: the long period of protein sparing during which energy expenditure is derived from lipid oxidation (phase II), and the later phase characterized by a rise in body protein utilization and plasma corticosterone (phase III). Apoptosis was further studied in 2, 6, and 24 h refed rats. Morphological apoptotic events were observed by environmental and conventional scanning electron microscopy and a TUNEL test was used to characterize the final stages of apoptotic death. The gene and protein expressions of TNFalpha, TGFbeta1, and Cdx2 were measured. Apoptotic events and TNFalpha, TGFbeta1, and Cdx2 gene and protein expressions did not vary significantly during phase II as compared to the normally fed animals. However, a phase III fasting induced a delay in intestinal epithelial apoptosis, along with a 92, 58, and 25% decrease in TNFalpha, TGFbeta1, and Cdx2 mRNAs, respectively. The amounts of TNFalpha, TGFbeta1, and Cdx2 proteins decreased by 70, 36, and 25%, respectively. Apoptosis was restored rapidly after a 2 h refeeding following the phase III, accompanied by a significant increase in TNFalpha, TGFbeta1, and Cdx2 mRNA and the protein levels, compared to the phase III fasting values. The concomitant decreases in cytokines and Cdx2 and in apoptotic cells during phase III suggest the preservation of enterocytes during this critical fasting period in order to optimize nutrient absorption as soon as food is available and thus, to rapidly restore body mass

P73 Preliminary results of the Study of Acute Liver Transplant (SALT): NSAID exposure and risk of acute liver failure leading to transplantation in 7 European countries

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Behavioural and Physiological Responses of Gammarus pulex Exposed to Cadmium and Arsenate at Three Temperatures: Individual and Combined Effects

This study aimed at investigating both the individual and combined effects of cadmium (Cd) and arsenate (AsV) on the physiology and behaviour of the Crustacean Gammarus pulex at three temperatures (5, 10 and15°C). G. pulex was exposed during 96 h to (i) two [Cd] alone, (ii) two [AsV] alone, and (iii) four combinations of [Cd] and [AsV] to obtain a complete factorial plane. After exposure, survival, [AsV] or [Cd] in body tissues, behavioural (ventilatory and locomotor activities) and physiological responses (iono-regulation of [Na+] and [Cl−] in haemolymph) were examined. The interactive effects (antagonistic, additive or synergistic) of binary mixtures were evaluated for each tested temperature using a predictive model for the theoretically expected interactive effect of chemicals. In single metal exposure, both the internal metal concentration in body tissues and the mortality rate increased along metallic gradient concentration. Cd alone significantly impaired both [Na+] and [Cl−] while AsV alone had a weak impact only on [Cl−]. The behavioural responses of G. pulex declined with increasing metal concentration suggesting a reallocation of energy from behavioural responses to maintenance functions. The interaction between AsV and Cd was considered as ‘additive’ for all the tested binary mixtures and temperatures (except for the lowest combination at 10°C considered as “antagonistic”). In binary mixtures, the decrease in both ventilatory and locomotor activities and the decline in haemolymphatic [Cl−] were amplified when respectively compared to those observed with the same concentrations of AsV or Cd alone. However, the presence of AsV decreased the haemolymphatic [Na+] loss when G. pulex was exposed to the lowest Cd concentration. Finally, the observed physiological and behavioural effects (except ventilation) in G. pulex exposed to AsV and/or Cd were exacerbated under the highest temperature. The discussion encompasses both the toxicity mechanisms of these metals and their interaction with rising temperature

The comparative osmoregulatory ability of two water beetle genera whose species span the fresh-hypersaline gradient in inland waters (Coleoptera: Dytiscidae, Hydrophilidae).

A better knowledge of the physiological basis of salinity tolerance is essential to understanding the ecology and evolutionary history of organisms that have colonized inland saline waters. Coleoptera are amongst the most diverse macroinvertebrates in inland waters, including saline habitats; however, the osmoregulatory strategies they employ to deal with osmotic stress remain unexplored. Survival and haemolymph osmotic concentration at different salinities were examined in adults of eight aquatic beetle species which inhabit different parts of the fresh-hypersaline gradient. Studied species belong to two unrelated genera which have invaded saline waters independently from freshwater ancestors; Nebrioporus (Dytiscidae) and Enochrus (Hydrophilidae). Their osmoregulatory strategy (osmoconformity or osmoregulation) was identified and osmotic capacity (the osmotic gradient between the animal's haemolymph and the external medium) was compared between species pairs co-habiting similar salinities in nature. We show that osmoregulatory capacity, rather than osmoconformity, has evolved independently in these different lineages. All species hyperegulated their haemolymph osmotic concentration in diluted waters; those living in fresh or low-salinity waters were unable to hyporegulate and survive in hyperosmotic media (> 340 mosmol kg(-1)). In contrast, the species which inhabit the hypo-hypersaline habitats were effective hyporegulators, maintaining their haemolymph osmolality within narrow limits (ca. 300 mosmol kg(-1)) across a wide range of external concentrations. The hypersaline species N. ceresyi and E. jesusarribasi tolerated conductivities up to 140 and 180 mS cm(-1), respectively, and maintained osmotic gradients over 3500 mosmol kg(-1), comparable to those of the most effective insect osmoregulators known to date. Syntopic species of both genera showed similar osmotic capacities and in general, osmotic responses correlated well with upper salinity levels occupied by individual species in nature. Therefore, osmoregulatory capacity may mediate habitat segregation amongst congeners across the salinity gradient

Effects of tributyltin oxide on survival and osmoregulation of the shrimp Penaeus japonicus (crustacea, decapoda)

International audienceThe acute toxicity of tributyltin oxide (TBTO) was determined in larvae (nauplii, zoeae 1-3, mysis 1-3), post-larvae (PL stages) and juvenile shrimp (Penaeus japonicus Bate), in two media, seawater (SW) and diluted seawater (DSW; 1100 and 550 mosm kg-1 ≃37 and 19%). Survival, osmoregulatory capacity and Na+-K+ ATPase activity were measured. A gill and epipodite histopathological study was also conducted. The 24 and 48 h LC50s values for TBTO in SW ranged from 2.03 μg l-1 (1.7-2.4) and 0.88 μg l-1 (0.8- 1.0) for nauplii to 773 μg l-1 (344-1823) and 708 μg l-1 (361-1608) for juveniles. The 96 h LC50s values in SW ranged from 19.4 μg-1 (12.6- 27.3) for PL5 to 370 μg l-1 (202-662) for juveniles. The 96 h LC50s value was not affected by salinity in juveniles. Tolerance to TBTO tended therefore to increase with the development from larval to juvenile instars. In juveniles kept in SW and in DSW, acute TBTO-exposures decreased the osmoregulatory capacity (OC = difference between the hemolymph osmolality and the osmolality of the medium) of animals exposed to lethal and sublethal concentrations. Effects of TBTO exposure on hypo- and hyper-OC were time- and dose-dependent and the ability to osmoregulate was recovered after exposure of the shrimp to water free of TBTO for 48 120 h. These experiments confirmed OC as a valuable tool for monitoring the physiological state of peneid shrimp. Gill and epipodite Na+-K+ ATPase activities were not altered in SW and DSW after acute TBTO-exposures, either at sublethal or at lethal concentrations. Haemocytic congestion (thrombosis), multiple necrosis and nephrocyte hyperplasia were observed in gill lamellae of exposed shrimp. Multiple necrosis and lacunae in the epithelial monolayers were also observed in epipodites. At lethal concentrations, the interconnecting lacunae were reduced and/or replaced by proliferating tissues. Epithelial cells were peeling and oedema was observed. For both tissues, histopathological effects increased with the dose and they are probably the cause of impaired osmoregulation

Immunolocalisation of aquaporin 3 in the gill and the gastrointestinal tract of the European eel <em>Anguilla anguilla</em> (L.)

The expression of a putative water channel protein, aquaporin 3 (AQP-3), has been localised within branchial and intestinal tissues from the 'silver' life stage of the European eel Anguilla anguilla, using a specific polyclonal antibody directed against the C-terminal of the amino acid sequence. Western blots using the AQP-3 antiserum identified the presence of a major immunoreactive protein of 24 kDa in extracts of gills from both freshwater (FW) and 3 week seawater (SW)-acclimated eels. SW acclimation induced a 65% reduction in AQP-3 protein abundance in the gill extracts. AQP-3 immunoreactivity was apparent throughout the branchial epithelium from both FW and SW-acclimated fish, but especially so within the chloride cells, which also stained heavily with specific antisera for the beta-subunit of the Na, K-ATPase. AQP-3 immunoreactivity not only colocalised with Na, K-ATPase within the basolateral tubular network but also stained the apical regions of the chloride cell where Na, K-ATPase was absent. Although there were no obvious differences in expression between the chloride cells of FW and SW-acclimated fish, considerably higher intensities of immunoreactivity were apparent near the periphery of the non-chloride cells of FW fish, especially within cells forming the base of the primary filaments and the branchial arch. AQP-3 inummoreactivity was also detected in intra-epithelial macrophage-like cells within the intestine of FW and SW-acclimated eels and in the mucous cells of the rectal epithelium of SW-acclimated fish. These results suggest that AQP-3 may play an important functional role in osmoregulation the teleostean gill but is unlikely to be responsible for the increases in intestinal water absorption that occur following SW acclimation.</p