Three-dimensional imaging to estimate in vivo body and carcass chemical composition of growing beef-on-dairy crossbred bulls.

The dynamics of cattle body chemical composition during growth and fattening periods determine animal performance and beef carcass quality. The aim of this study was to estimate the empty body (EB) and carcass chemical composition of growing beef-on-dairy crossbred bulls (Brown Swiss breed as dam with Angus, Limousin or Simmental as sire) using three-dimensional (3D) imaging. The 3D images of the cattle's external body shape were recorded in vivo on 48 bulls along growth trajectory (75-520 kg BW and 34-306 kg hot carcass weight [HCW]; set 1) and on 70 bulls at target market slaughter weight, including 18 animals from set 1 (average 517 ± 10 kg BW and 289 ± 10 kg HCW; set 2). The linear, circumference, curve, surface and volume measurements on the 3D body shape were determined. Those predictive variables were used in partial least square regressions, together with the effect of the sire breed whenever significant (P < 0.05), with leave-one-out cross-validation to estimate water, lipid, protein, mineral and energy mass or proportions in the EB and carcass. Mass and proportions were determined directly from postmortem grinding and chemical analyses (set 1) or indirectly using the 11th rib dissection method (set 2). In set 1, bulls' BW and HCW were estimated via 3D imaging, with root mean square error of prediction (RMSEP) of 12 kg and 6 kg, respectively. The EB and carcass chemical component proportions were estimated with RMSEP from 0.2% for EB minerals (observed mean 3.7 ± 0.2%) to 1.8% for EB lipid (11.6 ± 4.2%), close to the RMSEP found for the carcass. In set 2, the RMSEP for estimation via 3D imaging was 9 kg for BW and 6 kg for HCW. The EB energy and protein proportions were estimated, with RMSEP of 0.5 MJ/kg fresh matter (10.1 ± 0.8 MJ/DM) and 0.2% (18.7 ± 0.7%), respectively. Overall, the estimations of chemical component proportions from 3D imaging were slightly less precise for both sets than the mass estimations. The morphological traits from the 3D images appeared to be precise estimators of BW, HCW as well as EB and carcass chemical component masses and proportions.

Assessment of chloride natural background levels by applying statistical approaches. Analyses of European coastal aquifers in different environments.

Estimated natural background levels (NBLs) are needed to assess groundwater chemical status according to the EU Groundwater Directive. They are commonly derived for different substances by applying statistical methodologies. Due to the complexity of the sea water intrusion process, some of those methods do not always provide appropriate assessment of chloride NBLs. This paper analyzes the applicability of different NBL estimation methods in five EU coastal aquifers with significant differences in available datasets and hydrogeological settings. A sensitivity analysis of results to different constraints was performed to remove samples with anthropogenic impacts. A novel statistical approach combining different methods to identify the range of chloride NBLs is proposed. In all pilots the estimated NBLs were below 85 mg/L and fitted well with previous studies and expert judgment, except Campina del Faro aquifer (the maximum being 167.5 mg/L). Although this approach is more time consuming, it provides a more robust solution.

[History of Ketamine: An ancient molecule that is still popular today]. / Histoire de la kétamine : une molécule ancienne qui a toujours la cote.

The history of ketamine begins in 1962, when Calvin Stevens of the pharmaceutical laboratory Parke-Davis synthesizes it from phencyclidine, a molecule with psychodysleptic, hallucinogenic and dissociative properties. Following the first administration of ketamine to humans in 1964 in Jackson prison (Michigan, USA), its dissociative effects associated with short anaesthesia were reported, and a patent for its human use was filed in 1966. In the 1990s, the discovery of opioid-induced hyperalgesia sparked interest in ketamine as an analgesic. In recent years, the human use of ketamine, and in particular its esketamine enantiomer, has shifted towards the treatment of depression. The first cases of ketamine abuse were reported in 1992 in France, leading to special surveillance by the health authorities, and its inclusion in the list of narcotic drugs in 1997. Today, ketamine has become an attractive substance for recreational use, gradually emerging from alternative techno circles to spread to more commercial party scenes. These elements represent a public health concern, associated with the risk of developing new chemically synthesized analogues, the harmful effects of which are still little known.

Clinical and biological assessment of lamotrigine and levetiracetam plasma assays at the Rennes University Hospital.

INTRODUCTION: Requests for lamotrigine and levetiracetam plasma assays have increased significantly since their development in the biological and forensic toxicology laboratory at the University Hospital of Rennes in 2015. The purpose of this study was to evaluate the follow-up of French National Authority for Health (HAS) guidelines for antiepileptic drug assays and the impact of assay results on medical management. METHODS: Two hundred and forty-two assay results of these two antiepileptics for 169 patients hospitalized in different care wards between 2015 and 2018 were retrospectively analyzed. RESULTS: The mean age of the study population was 50.3±25.4 years. Of the 207 assays prescribed for epilepsy, 177 (85.5%) were in line with the 2007 HAS guidelines, namely: 76/177 (42.9%) for therapeutic adjustment in the event of seizure recurrence or aggravation; 45/177 (25.4%) for specific clinical situations; 23/177 (13%) for proven or suspected poor compliance; 23/177 (13%) for suspected overdose; 8/177 (4.5%) following initiation of treatment; and 2/177 (1.1%) for drug interaction management. Thirty of the 207 assays (14.5%) were thus inappropriate. No significant differences were found regarding the hospitalization frequency after a visit to the emergency room (P=0.9) between patients with lamotrigine and/or levetiracetam plasma assays in therapeutic ranges versus those with concentrations outside the therapeutic ranges. Dosage changes were more frequent in patients with assays in therapeutic ranges compared to patients with plasma assays outside the therapeutic ranges (P=0.0015), suggesting a treatment reassessment primarily based on clinical criteria. CONCLUSION: The analysis of requests for antiepileptic drug assays at the University Hospital of Rennes revealed that clinicians were well aware of the HAS guidelines. In addition, the assay results were mainly consistent with clinical intuition, suggesting a real added value for patient management. However, the consequences in terms of changes in medical care seem limited. This assessment illustrates the importance of strengthening the dialogue between pharmacists, biologists and clinicians.

Effects of different types of solid feeds on health status and performance of Swiss veal calves. II. Basic feeding with whole milk.

The objective of this study was to identify a suitable alternative to the current practice of complementing the feeding of whole milk with straw. The influence of 3 different solid supplements on the health and performance of Swiss veal calves was investigated during 3 production cycles of 90 veal calves each with a mean initial age of 42 days and a mean initial weight of 68.7 kg. The calves were housed in groups of 30 in stalls strewn with wheat straw without outside pen. Liquid feeding consisted of whole milk combined with an additional skim milk powder ad libitum. Groups were assigned to one of the three following experimental solid feeds provided ad libitum: Pellet mix (composition: oat hulls, corn [whole plant], barley, sunflower seeds, squeezed grains of corn, molasses and a pellet binder), whole plant corn pellets, and wheat straw as control. Calves of the straw group showed significantly more abomasal lesions in the fundic part as compared to the pellet mix and corn pellets groups (P < 0.001), the prevalence of insufficient papillae was highest (P < 0.05), and ruminating behavior was unsatisfactory. In contrast to the pellet mix and straw groups, performance of calves in the corn pellets group was good. Additionally, prevalence of abomasal fundic lesions was lowest (P < 0.001), and rumen development was best in calves of the corn pellets group (P < 0.01). As in part I, the results reveal that whole-plant corn pellets are most consistent with an optimal result combining the calves' health and fattening performance. Therefore, it can be recommended as a solid supplement for veal calves basically fed whole milk under Swiss conditions.

Le but de cette étude était de trouver une alternative adéquate à la paille utilisée actuellement en tant qu'adjonction au lait entier. Pour cela, on a étudié l'effet de trois aliments solides différents sur la santé et la productivité de veaux suisses durant 3 périodes d'engraissement avec à chaque fois 90 veaux, une durée d'engraissement moyenne de 42 jours et un poids moyen de 68.7 kg au début de l'engraissement. Les veaux ont été détenus par groupes de 30 dans des écuries paillées sans enclos extérieur. L'alimentation liquide ad libitum se composait de lait entier complété par de la poudre de lait maigre. Les groupes ont reçu aléatoirement l'un des trois aliments solides testés, à savoir : cubes mélangés (composés de son d'avoine, de maïs plante entière, de graines de tournesol, de maïs aplati, de mélasse et de LignoBond DD comme agglomérant), cubes de maïs plante entière ou paille (groupe de contrôle). Les veaux du groupe «paille¼ présentait, comparativement à ceux des deux autres groupes, significativement plus de lésions de la caillette dans la zone du fundus (< 0.001), le nombre de veaux avec des villosités de la panse insuffisamment développées était plus élevé (P < 0.05) et la rumination était insatisfaisante. Contrairement à ceux des groupes «cubes mélangés¼ et «paille¼, les performances des veaux complémentés avec des cubes de maïs étaient bonnes et leurs carcasses correspondaient au mieux à la demande du marché. En outre, la prévalence de lésions de la caillette dans la zone du fundus était la plus basse chez les veaux nourris avec des cubes e maïs (P < 0.001) et le développement de la panse était le meilleur (P < 0.01). Comme dans la partie I de cette étude, les résultats présentés indiquent que les cubes de maïs plante entière sont une alternative applicable à l'affouragement de paille tel que pratiqué jusqu'à maintenant pour les veaux d'engrais.

Modulation of ethanol effect on hepatocyte proliferation by polyamines.

An occurrence and a magnitude of alcoholic liver diseases depend on the balance between ethanol-induced injury and liver regeneration. Like ethanol, polyamines including putrescine, spermidine, and spermine modulate cell proliferation. Thus, the purpose of this study was to evaluate the relationship between effect of ethanol on hepatocyte (HC) proliferation and polyamine metabolism using the HepaRG cell model. Results showed that ethanol effect in proliferating HepaRG cells was associated with a decrease in intracellular polyamine levels and ornithine decarboxylase (ODC) activity. Ethanol also induced disorders in expression of genes coding for polyamine-metabolizing enzymes. The α-difluoromethyl ornithine, an irreversible inhibitor of ODC, amplified ethanol toxicity on cell viability, protein level, and DNA synthesis through accentuation of polyamine depletion in proliferating HepaRG cells. Conversely, putrescine reversed ethanol effect on cell proliferation parameters. In conclusion, this study suggested that ethanol effect on HC proliferation was closely related to polyamine metabolism and that manipulation of this metabolism by putrescine could protect against the anti-proliferative activity of ethanol.

Metabolic and energy status during the dry period is crucial for the resumption of ovarian activity postpartum in dairy cows.

It is well known that the degree of negative energy balance in high-producing dairy cows is the major cause of delayed resumption of the ovarian cyclicity that closely relates to fertility. Recent evidence suggests that the energetic situation during early lactation critically affects nutrient partitioning, metabolism, and the reproductive axis, whereas the effect of energy status during the dry period is widely unknown. The aim of the present study was to investigate the effect of energy status throughout the entire dry period until early lactation on the onset of the ovarian cyclicity. Blood samples were taken in 23 cows from dry off at 8 wk before expected parturition to 8 wk postpartum for the analyses of metabolites and hormones, and milk samples were obtained 3 times weekly from d 7 of lactation onward to confirm luteal activity and pregnancy by milk progesterone analysis. Energy balance (EB) was measured weekly during the last 6 wk of the dry period and every other week after parturition. Liver biopsies were obtained at 8 wk before expected calving, within 1 d after calving, and at 4 wk postpartum to measure the mRNA abundance of various gluconeogenic enzymes and metabolic hormone receptors. Cows showing luteal activity within 3 wk postpartum were defined as ovulatory during the first follicular wave postpartum (OC), whereas cows without luteal activity within 3 wk postpartum were defined as anovulatory (AC). Energy balance and, concomitantly, plasma concentrations of glucose, insulin, insulin-like growth factor-1, 3,5,3'-triiodothyronine, and thyroxine were higher in OC than in AC during the dry period. Plasma thyroxine concentrations and body condition score during the postpartum period were higher in OC than in AC. At the mRNA level (19 cows), hepatic insulin receptor decreased from dry off to early lactation, and mRNA of pyruvate carboxylase was highest at parturition and decreased in early lactation in AC only, whereas both parameters remained unchanged in OC. The mRNA abundance of phosphoenolpyruvate carboxykinase-mitochondrial increased from dry off to parturition in both groups, remained high in OC, and decreased again in early lactation in AC. However, none of the investigated gene transcripts differed between OC and AC cows. Thus, ovarian function postpartum appears to be crucially influenced by the energy status during the dry period, which is reflected by timely changes in hepatic mRNA abundance of only a few key metabolic factors in the liver.

Coordinated gene expression in adipose tissue and liver differs between cows with high or low NEFA concentrations in early lactation.

Dairy cows with high and low plasma non-esterified fatty acid (NEFA) concentrations in early lactation were compared for plasma parameters and mRNA expression of genes in liver and subcutaneous adipose tissue. The study involved 16 multiparous dairy cows with a plasma NEFA concentration of >500 µmol/l [n = 8, high NEFA (HNEFA)] and <140 µmol/l [n = 8, low NEFA (LNEFA)] in the first week post-partum (pp). Blood samples, adipose and liver tissues were collected on day 1 (+1d) and at week 3 pp (+3wk). Blood plasma was assayed for concentrations of metabolites and hormones. Subcutaneous adipose and liver tissues were analysed for mRNA abundance by real-time qRT-PCR encoding parameters related to lipid metabolism. Results showed that mean daily milk yield and milk fat quantity were higher in HNEFA than in LNEFA cows (p < 0.01), and the NEB was more negative in HNEFA than in LNEFA in +3wk too (p < 0.05). HNEFA cows had slightly lower (p < 0.1) insulin concentrations than LNEFA cows across the study period, and the body condition score decreased more from +1d to +3wk in HNEFA than in LNEFA (p = 0.09). The mRNA abundance of genes in the liver related to fatty acid oxidation (carnitine palmitoyltransferase 2 and very long chain acyl-coenzyme A dehydrogenase) and ketogenesis (3-hydroxy-3-methylglutaryl-coenzyme A synthase 2) were lower in HNEFA than in LNEFA cows. No differences between the two groups were observed for mRNA expression of genes in adipose tissue. The number of calculated significant correlation coefficients (moderately strong) between parameters in the liver and in adipose tissue was nearly similar on +1d, and higher for HNEFA compared with LNEFA cows in +3wk. In conclusion, dairy cows with high compared with low plasma NEFA concentrations in early lactation show differentially synchronized mRNA expression of genes in adipose tissue and liver in +3wk that suggests a different orchestrated homeorhetic regulation of lipid metabolism.

Plasma leptin and mRNA expression of lipogenesis and lipolysis-related factors in bovine adipose tissue around parturition.

The objective was to study changes in plasma leptin concentration parallel to changes in the gene expression of lipogenic- and lipolytic-related genes in adipose tissue of dairy cows around parturition. Subcutaneous fat biopsies were taken from 27 dairy cows in week 8 antepartum (a.p.), on day 1 postpartum (p.p.) and in week 5 p.p. Blood samples were assayed for concentrations of leptin and non-esterified fatty acids (NEFA). Subcutaneous adipose tissue was analysed for mRNA abundance by real-time qRT-PCR encoding for leptin, adiponectin receptor 1 (AdipoR1), adiponectin receptor 2 (AdipoR2), hormones-sensitive lipase (HSL), perilipin (PLIN), lipoprotein lipase (LPL), acyl-CoA synthase long-chain family member 1 (ACSL1), acetyl-CoA carboxylase (ACC), fatty acid synthase (FASN) and glycerol-3-phosphate dehydrogenase 2 (GPD2). Body weight and body condition score of the cows were lower after parturition than before parturition. The calculated energy balance was negative in week 1 and 5 p.p., with higher negative energy balance in week 1 p.p. compared with that in week 5 p.p. On day 1 p.p., highest concentrations of NEFA (353.3 µmol/l) were detected compared with the other biopsy time-points (210.6 and 107.7 µmol/l, in week 8 a.p., and week 5 p.p. respectively). Reduced plasma concentrations of leptin during p.p. when compared with a.p. would favour increasing metabolic efficiency and energy conservation for mammary function and reconstitution of body reserves. Lower mRNA abundance of ACC and FASN expression on day 1 p.p. compared with other biopsy time-points suggests an attenuation of fatty acid synthesis in subcutaneous adipose tissue shortly after parturition. Gene expression of AdipoR1, AdipoR2, HSL, PLIN, LPL, ACSL1 and GPD2 was unchanged over time.

Gene expression of tumour necrosis factor and insulin signalling-related factors in subcutaneous adipose tissue during the dry period and in early lactation in dairy cows.

Gene expression of adipose factors, which may be part of the mechanisms that underlie insulin sensitivity, were studied in dairy cows around parturition. Subcutaneous fat biopsies and blood samples were taken from 27 dairy cows in week 8 antepartum (a.p.), on day 1 postpartum (p.p.) and in week 5 p.p. In the adipose tissue samples, mRNA was quantified by real-time reverse transcription polymerase chain reaction for tumour necrosis factor alpha (TNFα), insulin-independent glucose transporter (GLUT1), insulin-responsive glucose transporter (GLUT4), insulin receptor, insulin receptor substrate 1 (IRS1), insulin receptor substrate 2 (IRS2), regulatory subunit of phosphatidylinositol-3 kinase (p85) and catalytic subunit of phosphatidylinositol-3 kinase. Blood plasma was assayed for concentrations of glucose, ß-hydroxybutyric acid, non-esterified fatty acids (NEFA) and insulin. Plasma parameters followed a pattern typically observed in dairy cows. Gene expression changes were observed, but there were no changes in TNFα concentrations, which may indicate its local involvement in catabolic adaptation of adipose tissue. Changes in GLUT4 and GLUT1 mRNA abundance may reflect their involvement in reduced insulin sensitivity and in sparing glucose for milk synthesis in early lactation. Unchanged gene expression of IRS1, IRS2 and p85 over time may imply a lack of their involvement in terms of insulin sensitivity dynamics. Alternatively, it may indicate that post-transcriptional modifications of these factors came into play and may have concealed an involvement.

Variation in hepatic regulation of metabolism during the dry period and in early lactation in dairy cows.

The purpose of this study was to investigate variations in hepatic regulation of metabolism during the dry period, after parturition, and in early lactation in dairy cows. For this evaluation, cows were divided into 2 groups based on the plasma concentration of beta-hydroxybutyric acid (BHBA) in wk 4 postpartum (PP; group HB, BHBA >0.75 mmol/L; group LB, BHBA <0.75 mmol/L, respectively). Liver biopsies were obtained from 28 cows at drying off (mean 59 +/- 8 d antepartum), on d 1, and in wk 4 and 14 PP. Blood samples were collected every 2 wk during this entire period. Liver samples were analyzed for mRNA abundance of genes related to carbohydrate metabolism (pyruvate carboxylase, PC; phosphoenolpyruvate carboxykinase, PEPCK; citrate synthase, CS), fatty acid biosynthesis (ATP citrate lyase, ACLY) and oxidation (acyl-CoA synthetase long-chain, ACSL; carnitine palmitoyltransferase 1A, CPT 1A; carnitine palmitoyltransferase 2, CPT 2; acyl-coenzyme A dehydrogenase very long chain, ACADVL), cholesterol biosynthesis (3-hydroxy-3-methylglutaryl-coenzyme A synthase 1, HMGCS1), ketogenesis (3-hydroxy-3-methylglutaryl-coenzyme A synthase 2, HMGCS2), and of genes encoding the transcription factors peroxisome proliferator-activated receptor alpha (PPARalpha), peroxisome proliferator-activated receptor gamma (PPARgamma), and sterol regulatory element binding factor 1 (SREBF1). Blood plasma was assayed for concentrations of glucose, BHBA, nonesterified fatty acids, cholesterol, triglycerides, insulin, insulin-like growth factor-I, and thyroid hormones. In both groups, plasma parameters followed a pattern usually observed in dairy cows. However, changes were moderate and the energy balance in cows turned positive in wk 7 PP for both groups. Additionally, the energy balance and milk yield were similar for both groups after parturition onwards. Significant group effects were found at drying off, when plasma concentrations of triglycerides were higher in LB than in HB, and in wk 4 PP, when plasma concentrations of glucose and IGF-I were lower in HB than in LB. Similarly, moderate changes in mRNA expression of hepatic genes between the different time points were observed, although HB cows showed more adaptive performance than LB cows based on changes in mRNA expression of PEPCKc, PEPCKm, CS, CPT 1A, CPT 2, and PPARalpha. Part of the variation measured in this study was explained by parity. Significant Spearman rank correlation coefficients between the variables were not similar at each time point and were not similar between the groups at each time point, suggesting that metabolic regulation differs between cows. In conclusion, metabolic regulation in dairy cows is a dynamic system, and differs obviously between cows at different metabolic stages related to parturition.

Sexual assault and MDMA: the distinction between consciousness and awareness when it comes to consent.

In France, the term chemical or drug-assisted submission is usually defined as the rendering of a person vulnerable by the surreptitious administration of an active substance with the purpose of prejudicing the person or his/her possessions. If the harm is sexual assault, establishing the victim's submission involves both proving that a dangerous substance was administered, providing material evidence of the infraction (the assault), i.e. the detection of traces in a physical examination and samples, and proving the absence of consent. We report the case of a woman who was sexually assaulted after having been surreptitiously administered methylenedioxymethamphetamine. In this special case, the woman remained in a state of vigilance (conscious) throughout, so there is doubt about whether or not she consented. In other words, the ability to consent is debatable.

[Case of death occurred after an injection of aetoxisclerol. The responsibility of the product should be discussed]. / L'aetoxisclerol accusé à tort d'entraîner une mort subite.

A 35-year-old woman has suffered from faintness with cardiovascular failure at the end of a sclerotherapy session for varix management. The injected product was Aeloxisclerol (DCI lauromacrogal 400). The death came up very quickly. The autopsy didn't reveal any traumatic lesion. The anaphylactic choc could be ruled out. Macroscopically, the heart showed a right ventricular dilatation. The toxicological analysis didn't reveal any medicinal substances. Histology showed the presence of lipid degeneration of all the right-ventricle wall The discussion is carried out on the role played by this previous health-state on the onset of death, and also on the incidence of sclerotherapy using this product. and the onset of death. We are carrying out this discussion by taking in mind the nature of the product injected and the information available in the literature concerning the lipid degeneration of cardiac muscle. Quickly after this case, the AFSSAPS (The French Agency for Sanitary Security of Health Products) has emitted an written alert which correlates with the requirements of principles of precaution although it is not founded on the totality of investigation results done for this medicolegal case.

[Evaluation of portable point of care instrument: the i-Stat . Report of 7,000 analyses]. / Evaluation d'un analyseur de sant portable: l'i-STAT. Bilan de 7000 analyses.

To validate the use of the i-STAT portable point-of-care instrument, performance checks have been conducted to verify its accuracy for PCO2/PO2 (tonometry) and precision for pH/PCO2/PO2 (control solutions). Results obtained with the i-STAT (pH/PCO2/PO2/Hct) were also compared to those provided by the Bayer Rapidlab 865 routinely used in our laboratory. All these measurements have been conducted under the same conditions using three types of i-STAT cartridges (G3, EG6, EG7) with two of different life duration to each cartridge with a view to observing their potential variability between types and any effect that ageing may have on analytical performances. Differences observed in this respect are not clear enough to draw firm conclusions. On the other hand, we consider the level of PCO2 and particularly PO2 performances not sufficient to warranty their use in the absence of a very strict quality control coupled with a highly developed clinical sense. No real use can be made of values noted for hematocrit (1998 survey).

Free radical scavenging and antioxidant effects of lactate ion: an in vitro study.

Divergent literature data are found concerning the effect of lactate on free radical production during exercise. To clarify this point, we tested the pro- or antioxidant effect of lactate ion in vitro at different concentrations using three methods: 1) electron paramagnetic resonance (EPR) was used to study the scavenging ability of lactate toward the superoxide aion (O(2)(-).) and hydroxyl radical (.OH); 2) linoleic acid micelles were employed to investigate the lipid radical scavenging capacity of lactate; and 3) primary rat hepatocyte culture was used to study the inhibition of membrane lipid peroxidation by lactate. EPR experiments exhibited scavenging activities of lactate toward both O(2)(-). and.OH; lactate was also able to inhibit lipid peroxidation of hepatocyte culture. Both effects of lactate were concentration dependent. However, no inhibition of lipid peroxidation by lactate was observed in the micelle model. These results suggested that lactate ion may prevent lipid peroxidation by scavenging free radicals such as O(2)(-). and.OH but not lipid radicals. Thus lactate ion might be considered as a potential antioxidant agent.

Activated macrophages increase the susceptibility of rat hepatocytes to ethanol-induced oxidative stress: conflicting effects of nitric oxide.

The aim of this study was to examine how macrophages could act on ethanol-induced oxidative stress in rat hepatocytes during inflammatory conditions, well-known to induce nitric oxide (NO) synthase. For this purpose, RAW 264.7 macrophages were added to primary rat hepatocyte cultures. Co-cultures were then supplemented with lipopolysaccharide (LPS) and interferon gamma (IFN) for 18 h, in order to induce NO synthase before the addition of 50 mM ethanol. In cultures of hepatocytes alone, the addition of LPS and IFN protected from ethanol-induced oxidative stress. It has been shown previously that NO generated in hepatocytes was responsible for this effect. When macrophages were added to primary rat hepatocyte cultures supplemented with LPS and IFN, protection provided by NO against ethanol-induced oxidative stress in hepatocytes ceased. Using a pretreatment of macrophages with N(g)-monomethyl-l-arginine, a NO synthase inhibitor, it was concluded that NO generated by macrophages was responsible for macrophage toxicity. Taken together, our observations suggest that NO biosynthesis in hepatocytes protects them from ethanol-induced oxidative stress, whereas NO production in macrophages deprives hepatocytes of this NO protection.

Irniine, a pyrrolidine alkaloid, isolated from Arisarum vulgare can induce apoptosis and/or necrosis in rat hepatocyte cultures.

The effects of irniine, a pyrrolidine alkaloid extracted from the tubers of Arisarum vulgare, on rat hepatocyte primary cultures and rat liver epithelial cell line (RLEC) were studied. Cytotoxicity was first evaluated by LDH release, MTT and NR tests and MDA production, while cellular alterations were visualized by electron microscopy and DNA gel-electrophoresis. In hepatocyte and RLEC cultures, a major toxicity appeared at 40 microM of irniine and was demonstrated by an increase in LDH release and decreases in MTT reduction and NR uptake while concentrations lower than 40 microM did not induce significant changes in these parameters. However, we observed an increase in MDA production at 30 microM. Important alterations of the nuclei and mitochondria were also visualized by electron microscopy in cells treated with 50 microM. Using DNA gel-electrophoresis, we demonstrated that irniine at 40 and 50 microM induced DNA damage. All together these results demonstrate that: (1) Irniine induces a significant hepatotoxicity. (2) Irniine toxicity is not mediated by a metabolic derivative since RLEC, which do not contain a monooxygenase system, were also affected by this compound. (3) Irniine induces a significant DNA damage and oxidative stress which leads to cell death by necrosis and/or by apoptosis. Moreover, our data suggest that the alkaloid irniine contained in A. vulgare may be involved in the toxic symptoms observed after medicinal use or consumption of the plant tubers as food both by humans and animals.

Cytoprotection and iron mobilization in rat hepatocyte cultures by a new synthetic dihydroxamate chelator.

The cytoprotection and iron mobilization effect of a new dihydroxamate chelator 1,1 bis [(11-N-hydroxy)-2,5,11-triaza-1,6,10-trioxo dodecanyl] ethane or KD was studied in primary rat hepatocyte cultures exposed to iron-citrate. Lactate dehydrogenase (LDH) release and malondialdehyde (MDA) production were measured as indexes of cytotoxicity. Cell viability was evaluated using the [3-(4,5-dimethyl-thiazol-2-yl) 2,5-diphenyl tetrazolium bromide] (MTT) reduction test. To demonstrate that this chelator was able to decrease iron uptake or increase iron release from the hepatocytes, labelled cells were obtained by maintaining the cultures in the presence of 0.02 microM 55Fe-citrate. The efficacy of KD was compared to desferrioxamine B (DFO) at stoechiometry concentrations. After 24 h of exposure to 50 microM of iron-citrate, a significant release of LDH and MDA was observed. Cell viability was also significantly decreased. When 100 microM of KD were added at the same time as iron, LDH and MDA release was decreased and cell viability was improved. In the presence of the same chelator concentration, a net decrease of iron uptake by the cells was observed as attested by the low intracellular 55Fe level. Moreover, in the 55Fe loaded hepatocytes, the chelator increased the iron extracellular level indicating its iron release effect from the cells. In all tested experimental conditions, the efficacy of 100 microM of the dihydroxamate chelator KD was close to that of 50 microM of the trihydroxamate chelator DFO. In conclusion, KD is effective at a level comparable to DFO in protecting rat hepatocytes against the toxic effect of iron-citrate by decreasing the uptake of the metal and increasing its release from the cells. This synthetic compound appears to have some potential therapeutical interest and the results obtained encourage the synthesis of new hydroxamate ligands.