Modular bioreactor for primary human hepatocyte culture: medium flow stimulates expression and activity of detoxification genes

Down-regulation of detoxification genes, notably cytochrome P450 (CYPs), in primary hepatocyte cultures is a long-standing and major concern. We evaluated the influence of medium flow in this model. Hepatocytes isolated from 12 different liver donors were cultured either in a multichamber modular bioreactor (MCmB, flow rate 250-500 μL/min) or under standard/static conditions, and the expression of 32 genes, enzyme activities and biological parameters were measured 7-21 days later. mRNA expression of genes involved in xenobiotic/drug metabolism and transport, including CYP1A1, 1A2, 2B6, 2C9, 3A4 (and activities for some of them), UDP-glucuronosyltransferase (UGT) 1A1, UGT2B4, UGT2B7, glutathione S-transferase (GSTα), and multidrug resistance protein 1 (MDR1) and MRP2, were specifically up-regulated by medium flow as compared with static controls in all cultures tested. In 2-week-old cultures, expression of detoxification genes reached levels close to or higher than those measured in freshly isolated hepatocytes. In contrast, CYP2D6 and most of other tested genes were not affected by medium flow. We conclude that medium flow specifically interferes with, and up-regulates, the activity of xenosensors and/or the expression of detoxification genes in primary human hepatocytes. Down-regulation of detoxification genes in conventional (static) cultures is therefore partly a consequence of the absence of medium circulation.     

Le Plio-Pléistocène de Camerota(Italie méridionale): Géomorphologie et Paléoflores

Geomorphological investigations have been carried out in Camerota surroundings where marine microfaunas have been discovered. So, we can separate the Late Pliocene deposits from Calabrian and post-Calabrian levels. Preliminary palynological results show a vegetational evolution in relation with the climate. Leaf-remains give informations on the structure of the vegetation.     

Climatic inference from the history of the taxodiaceae during the pliocene and the early pleistocene in Western Europe

The Taxodiaceae are present throughout Western Europe during the Early Pliocene. Chronological data show that their extinction begins during the Middle Pliocene in the French Mediterranean area and that it occurs later (during the Late Pliocene and Early Pleistocene) in northwestern Europe, in elevated regions and in Italy. This chronology is difficult to understand if we accept cooling of the climate as the only determining factor of this extinction. The non-synchronous extinction of the Taxodiaceae in Western Europe reflects the more complex pattern of climatic phenomena which include variations in precipitation and temperature at different times and with differing intensity according the area. A decrease in precipitation forced the extinction of the Taxodiaceae in southeastern France (summer drought) and it occurred before the temperature threshold for Taxodiaceae was reached in the other parts of Europe.     

Renal immunolocalization of kallikrein in cisplatin nephrotoxicity in rats

Summary Treatment of rats with cisplatin (4 mg kg^-1body wt i.p. injection) induced variations of urinary kallikrein excretion (UKE). Three phases were observed: a transient increase of UKE one day after injection, followed by a decrease up to 10 days suggesting an altered biosynthesis and a recovery phase with return to normal control values, 21 days after injection. Early morphological lesions were observed in proximal tubule cells on day 1; severe changes and tubular necrosis were observed in the following days. Less marked changes were also present in distal tubules but the vacuolated and desquamated cells appeared in the lumen of the tubules. By immunocytochemical methods, kallikrein was observed in connecting tubule cells, but also in some proximal tubule cells and along the endothelial side of the glomerular basement membrane and urinary space of glomeruli. An intense labelling was present in desquamated epithelial cells in dilated lumen of tubules. This study provides evidence of the presence of immunoreactive kallikrein in the glomerulus, already reported during acute failure, and confirms the use of urinary kallikrein measurements as a useful non-invasive index to assess a possible nephrotoxic effect at the distal level.     

Response of Photochemical Processes of Photosynthesis to Dinitrogen Fixation in Soybean

Symbiotic N2 fixation activity brings about changes in the photochemical processes of photosynthesis in soybean (Glycine max L. Merr.). For a potential photochemical efficiency ([phi]Po) similar to that obtained with an exclusively mineral nutrition, soybean, at full bloom stage (R2) with a moderate N2 fixation activity, had a better electron transfer quantum yield ([phi]PSII) and a higher photochemical quenching. At the beginning seed stage (R5), corresponding to more intense N2 fixation, the same phenomena were enhanced; in addition, an effect on the photochemical (k2b) and nonphotochemical (Kn-k22) transfer rates and an earlier activation of the electron transfer chain were characterized using a new parameter, the relative induction time of PSII fluorescence (Ap/Fm). The response of the photochemical parameters was related to the N2 fixation level (performance of the host plant-microsymbiont association): the energetic cost of symbiotic N2 fixation appeared to be met by a better photochemical efficiency of photosynthesis coupled with a decrease in thermal dissipation (kn - k22), by faster thylakoid energization, and by faster reopening of photosystem II centers at the time of fluorescence induction, as shown by decreased Ap/Fm.     

Presence de pollens d'Hamamelis (Hamamelidaceae,Angiospermae) dans le pliocene du Sud de la France

Pollen-analysis shows the presence of the genus Hamamelis in the pliocene flora of southern France. It contributesto the knowledge of the past distribution of taxa which are now extinct in western Europe.     

Apports de la palynologie a la connaissance du Pliocène du Roussillon (sud de la France)

The present study illustrates clearly how pollen analysis may be applied to stratigraphy from a botanical point of view. Within a chronologically reliable frame (Middle Pliocene for the marine deposits on the basis of Foraminifera record; two subzones for continental deposits on the basis of Mammals record: Hautimagne for Terrats fauna, Sète for Serrat-d'en-Vacquer fauna), palynology provides a good stratigraphical boundary: the extinction of the Taxodiaceae. In a remblayage area, it is established that a continental level is not necessarily younger than a marine one unless they are superposed. Many profiles are replaced according to the «progradation of the pliocene gulf of Roussillon. The boundary between marine and continental deposits cuts through the chronological line of the Taxodiaceae extinction. The flora investigated (93 taxa) is the first known for the Pliocene of this area. The extinction of the Taxodiaceae in southern France has a climatic cause: the setting in of a mediterranean rhythm (dry summers). This extinction is much older than the one which took place in Italy (Tiberian boundary) and in the Netherlands (Reuverian-Pretiglian boundary).     

Localization of a renal kallikrein immunoreactive-like substance in rat ureter

An antibody against rat kallikrein was produced in rabbits and its localization was studied in various organs of the rat to confirm its specificity. The distribution of immunoreactive kallikrein was studied in rat ureter by use of immunochemical techniques. Ureteral tissue was fixed in Zamboni's-glutaraldehyde fixative and immunostained with indirect immunofluorescence and the peroxidase-antiperoxidase (PAP) method for light and electron microscopy. Preabsorption of the primary polyclonal antiserum with purified rat urinary kallikrein and substitution with normal serum were used as controls. By light microscopy, kallikrein was localized in the lamina propria and in the adventitial connective tissue surrounding the entire ureter. Immunoelectron microscopy confirmed this immunolocalization. Immunoreactive kallikrein was concentrated in fibroblasts of connective tissue and was not present in collagen fibers. Immunoreactivity was associated with the Golgi complex, free polyribosomes, and rough endoplasmic reticulum. No immunostaining was observed in other subcellular components of fibroblasts.     

Analyse pollinique des dépôts lacustresdu Pléistocène inférieur de Banyoles (Bañolas,site de la Bòbila Ordis - Espagne): un élément nouveau dans la reconstitution de l'histoire paléoclimatique des régions méditerranéennes d'Europe occidentale

Clays from Bòbila Ordis near Banyoles have yielded a rich pollen flora. Three pollen zones are recognizable: the lower and the middle parts are dominated by components of a deciduous forest (with numerous exotic taxa); in the upper part, Pinus pollen and NAP (especially Compositae) are in a majority with an increase of mediterranean taxa towards the top. This floristic evolution may be connected with a climatic change characterized by a decrease in humidity which is itself linked to a world cooling. We suggest a possible relation with the pollen flora published by H. Elhai. The pollen diagram of Bòbila Ordis is the second so far established from Lower Pleistocene deposits in the Mediterranean countries of Western Europe; chronologically it follows the Bernasso (Languedoc) pollen diagram and confirms the latter results.     

Histoire plio-pléistocène des écosystèmes végétaux de Méditerranée sud-occidentale : apport de l’analyse pollinique de deux sondages en mer d’Alboran

The pollen analysis of sediments of two boreholes located in the northeastern Morocco (Nador 1) and in the southeastern Spain (Andalucia G1) has allowed shedding a new light on the vegetation and climate of the North African littoral plains, the Rif Massif and the Betic Range during the Pliocene. The vegetation around the Alboran Sea was open and xeric during the Zanclean, dominated by herbs including subdesertic elements as Calligonum, Lygeum, Nitraria and Neurada. This type of vegetation indicates a dry and hot climate. The southwestern Mediterranean steppes have therefore a climatic character; they existed before the presence and the heavy pressure of Man on the environment. From the Piacenzian, the development, at Andalucia as well as at Nador, of Artemisia and the appearance of some altitude trees such as Cedrus and Cathaya indicate a vegetation change linked to a climatic change. Modifications in the vegetation observed during the same period in northwestern Mediterranean seem to indicate that the vegetation changes observed at Andalucia and Nador are controlled by the appearance of the first arctic glacial-interglacial cycles. The cedar tree appears at Nador only at the end of the Piacenzian, at time of the first cooling, while it has been found punctually in Messinian and early Pliocene sediments of Habibas in Algeria and in Messinian sediments of the Bou Regreg section at Salé in Morocco. The regular presence of Cedar at Bou Regreg during the Messinian, allows supposing that it was present in the Middle Atlas Mountains and that its development was favoured by colder conditions.