Nutritional regulation and kinetics of flocculosin synthesis by Pseudozyma flocculosa

This study sought to identify the factors and conditions that affected production of the antifungal glycolipid flocculosin by the biocontrol agent Pseudozyma flocculosa. For this purpose, different parameters known or reported to influence glycolipid release in fungi were tested. Concentration of the start-up inoculum was found to play an important role in flocculosin production, as the optimal level increased productivity by as much as tenfold. Carbon availability and nitrogen source (i.e., organic vs inorganic) both had a direct influence on the metabolism of P. flocculosa, leading to flocculosin synthesis. In general, if conditions were conducive for production of the glycolipid, carbon availability appeared to be the only limiting factor. On the other hand, if yeast extract was supplied as nitrogen source, fungal biomass was immediately stimulated to the detriment of flocculosin synthesis. Unlike other reports of glycolipid release by yeast-like fungi, inorganic nitrogen starvation did not trigger production of flocculosin. The relationship between the factors influencing flocculosin production in vitro and the conditions affecting the release of the molecule by P. flocculosa in its natural habitat appears to be linked to the availability of a suitable and plentiful food source for the biocontrol agent.     

Mitochondrial adaptations to steatohepatitis induced by a methionine- and choline-deficient diet

Nonalcoholic fatty liver disease (NAFLD) has become common liver disease in Western countries. There is accumulating evidence that mitochondria play a key role in NAFLD. Nevertheless, the mitochondrial consequences of steatohepatitis are still unknown. The bioenergetic changes induced in a methionine- and choline-deficient diet (MCDD) model of steatohepatitis were studied in rats. Liver mitochondria from MCDD rats exhibited a higher rate of oxidative phosphorylation with various substrates, a rise in cytochrome oxidase (COX) activity, and an increased content in cytochrome aa3. This higher oxidative activity was associated with a low efficiency of the oxidative phosphorylation (ATP/O, i.e., number of ATP synthesized/natom O consumed). Addition of a low concentration of cyanide, a specific COX inhibitor, restored the efficiency of mitochondria from MCDD rats back to the control level. Furthermore, the relation between respiratory rate and protonmotive force (in the nonphosphorylating state) was shifted to the left in mitochondria from MCDD rats, with or without cyanide. These results indicated that, in MCDD rats, mitochondrial ATP synthesis efficiency was decreased in relation to both proton pump slipping at the COX level and increased proton leak although the relative contribution of each phenomenon could not be discriminated. MCDD mitochondria also showed a low reactive oxygen species production and a high lipid oxidation potential. We conclude that, in MCDD-fed rats, liver mitochondria exhibit an energy wastage that may contribute to limit steatosis and oxidative stress in this model of steatohepatitis.     

Optimized adeno-associated virus 8 produces hepatocyte-specific Cre-mediated recombination without toxicity or affecting liver regeneration

Engineering viral vectors to produce liver-specific protein expression may help advance understanding of hepatic regeneration and disease states. In addition to introducing genes of interest to the liver, these vectors can be adapted for gene deletion when designed to express Cre recombinase. The ability to use this system requires high, liver-restricted expression, low toxicity, and no effect on the process of interest. We developed an adeno-associated virus 8 (AAV8) with a codon-optimized Cre recombinase under a hepatocyte-specific major urinary protein (MUP) promoter (MUP-iCre-AAV8) that fulfills these requirements. A single intravenous injection of ROSA26R reporter mice, which express lacZ after Cre-mediated recombination, demonstrated homogeneous beta-galactosidase expression limited to hepatocytes after only 7 days. Cre protein expression remained strong for at least 31 days. Serum liver function tests and histology demonstrated minimal liver toxicity. The presence of MUP-iCre-AAV8 did not affect hepatocyte proliferation after partial hepatectomy as measured by Ki67 staining. Conclusion: AAV8 with the MUP promoter, by virtue of its lack of hepatic toxicity or effect on liver regeneration, may be an efficient alternative to complex transgenic methodologies for studies of the mouse liver.     

PYY3-36 injection in mice produces an acute anorexigenic effect followed by a delayed orexigenic effect not observed with other anorexigenic gut hormones

Peptide YY (PYY) is secreted postprandially from the endocrine L cells of the gastrointestinal tract. PYY(3-36), the major circulating form of the peptide, is thought to reduce food intake in humans and rodents via high-affinity binding to the autoinhibitory neuropeptide Y (NPY) receptor within the arcuate nucleus. We studied the effect of early light-phase injection of PYY(3-36) on food intake in mice fasted for 0, 6, 12, 18, 24, and 30 h and show that PYY(3-36) produces an acute anorexigenic effect regardless of the duration of fasting. We also show evidence of a delayed orexigenic effect in ad libitum-fed mice injected with PYY(3-36) in the early light phase. This delayed orexigenic effect also occurs in mice administered a potent analog of PYY(3-36), d-Allo Ile(3) PYY(3-36), but not following injection of other anorectic agents (glucagon-like-peptide 1, oxyntomodulin, and lithium chloride). Early light-phase injection of PYY(3-36) to ad libitum-fed mice resulted in a trend toward increased levels of hypothalamic NPY and agouti-related peptide mRNA and a decrease in proopiomelanocortin mRNA at the beginning of the dark phase. Furthermore, plasma levels of ghrelin were increased significantly, and there was a trend toward decreased plasma PYY(3-36) levels at the beginning of the dark phase. These data indicate that PYY(3-36) injection results in an acute anorexigenic effect followed by a delayed orexigenic effect.     

The adventures of Sonic Hedgehog in development and repair. I. Hedgehog signaling in gastrointestinal development and disease

Hedgehog (Hh) proteins are members of a family of secreted signaling factors that orchestrate the development of many organs and tissues including those of the gastrointestinal (GI) tract. The requirement for Hh activity is not limited to early development but underlies the homeostasis of a number of tissues, and abnormal activity of the Hh pathway is associated with several GI malignancies. Understanding the roles and mechanisms of action of Hh signaling both in development and postnatally should thus give novel insights into potential treatments for these diseases. Here we focus on the Hh signaling pathway and its role in GI tract development and maintenance and consider the diseases resulting from aberrant Hh activity.     

Cationic liposomal lipids: from gene carriers to cell signaling

Cationic lipids are positively charged amphiphilic molecules which, for most of them, form positively charged liposomes, sometimes in combination with a neutral helper lipid. Such liposomes are mainly used as efficient DNA, RNA or protein carriers for gene therapy or immunization trials. Over the past decade, significant progress has been made in the understanding of the cellular pathways and mechanisms involved in lipoplex-mediated gene transfection but the interaction of cationic lipids with cell components and the consequences of such an interaction on cell physiology remains poorly described. The data reported in the present review provide evidence that cationic lipids are not just carriers for molecular delivery into cells but do modify cellular pathways and stimulate immune or anti-inflammatory responses. Considering the wide number of cationic lipids currently available and the variety of cellular components that could be involved, it is likely that only a few cationic lipid-dependent functions have been identified so far.     

Changes in social environment induce higher emotional disturbances than changes in physical environment in quail

In intensive rearing, birds are exposed to frequent changes in both their social and physical environments, and this can have an impact on animal welfare under commercial conditions. The aim of the present study was to compare the emotional responses induced by social and non-social changes and to study the influence of the familiarity on these responses.Twenty-two female quail were first reared with a ball in 15–20-individual groups for 3 weeks from hatching. Then, each experimental subject was allocated either a familiar congener (CONG) or a familiar object (BALL). At 6 weeks of age the birds were fitted with a telemetric device to collect motor and cardiac activities. After that, each subject was exposed first to a separation from its cage-mate/familiar object, and secondly to the (re-)introduction of either the cage-mate/familiar object or of an unknown conspecific/another ball. Emotional responses were assessed through behavioural reactions and heart rate variability.Before being separated, both BALL and CONG quail showed stereotyped pacing that was more pronounced in BALL quail. BALL quail were not affected by the ball withdrawal, unlike CONG quail which reacted to the separation from their cage-mate by reducing activity and exploratory behaviour (P < 0.05). After the re-introduction, BALL quail remained closer to their ball and CONG quail spent more time with “stretched necks” than before the separation (P < 0.05). Surprisingly, CONG quail showed less stereotyped pacing when their partner was removed and recovered the initial level of stereotyped behaviour when their cage-mate was re-introduced (P < 0.01). Likewise, CONG quail also recovered the initial values of heart rate after the re-introduction of their cage-mate, indicating an increase in sympathetic activity compared to the isolation period. When the unfamiliar congener or ball was introduced in their cage, BALL quail pecked the unknown ball more than the familiar ball and had more contact with it (P < 0.05) and CONG quail had more contact with the unknown congener than with the cage-mate (P < 0.05).In conclusion, the quail appeared to experience a negative affective state before being separated, and there was no clear evidence of negative emotion in quail in response to the separation from either a social partner or an inanimate object. Nevertheless, the emotional responses of the quail in reaction to the (re-)introduction was influenced by the familiarity of the congener or the ball. Finally, changes in the quail's social environment induced more behavioural and cardiac modifications than changes in its non-social environment.     

The effect of low energy electric shock on cortisol, β-endorphin, heart rate and behaviour of cattle

Electrical stimuli are used increasingly to confine cattle, whether through conventional electric fencing or the development of ‘virtual’ fencing systems. Two experiments were conducted to assess behavioural, heart rate and stress hormone responses of cattle to electrical stimuli typically used in such confinement applications. In the first experiment, 30 steers (18-months old; n = 10 per treatment) were held in a handling crush for 15 min after receiving one of the following treatments: nothing (control); delivery of three shocks at 2 s intervals (600 V, 250 mW); and restraint in a head bail for 3 min. Plasma cortisol and β-endorphin concentrations were measured at 0, 5, 10, 15 min, 1, 2, 3, and 4 h. In a second experiment, heart rate and behaviour were measured in 17 heifers (18 months of age) subjected to one of the following treatments whilst held in a crush for 10 min: nothing (control; n = 5); delivery of three shocks at 2 s intervals (600 V, 250 mW; n = 6); and restraint in a head bail for 3 min (n = 6). Cortisol and β-endorphin concentrations did not differ between treatments (P > 0.05). Whilst animals were receiving the treatments, heart rate was lower when head restrained compared with shock or control treatments (P = 0.009) and did not differ between control and electric shock treatments (P = 0.35). Upon release from the crush, heart rate was higher in shock and head restrained treatments than the control treatment (P = 0.005). Animals receiving the electric shock treatment tossed their heads more frequently whilst in the crush than control animals (P = 0.012) but did not differ from the other treatments in the number of vocalisations, tail swishes, steps back and forward, head tilts and head turns. There was a significant effect of treatment on flight time (P = 0.005); animals receiving the electric shocks were faster to leave the crush than control animals (P = 0.005) and there was no difference between head restraint and shock treatment (P = 0.86). In 10 min following release from the crush, there was no treatment difference in the time to start feeding. This study suggests that the stress response of cattle to low energy electric shocks is minimal and is similar to that induced by restraint in a crush.     

Consequences of ions and pH on the supramolecular organization of sphingomyelin and sphingomyelin/cholesterol bilayers

For drug delivery purpose the anticancer drug S12363 was loaded into ESM/Chol-liposomes using either a pH or an ammonium gradient. Association between the drug and the liposome depends markedly on the liposome membrane structure. Thus, ESM and ESM/Chol bilayer organization had been characterized by coupled DSC and XRDT as a function of both cholesterol concentration and aqueous medium composition. ESM bilayers exhibited a ripple lamellar gel phase P(beta') below the melting temperature and adopted a L(beta)-like gel phase upon Chol insertion. Supramolecular organization of ESM and ESM/Chol bilayers was not modified by citrate buffer or ammonium sulfate solution whatever the pH (3< or = pH < or =7). Nevertheless, in ESM bilayer, ammonium sulfate salt induced a peculiar organization of head groups, leading to irregular d-spacing and weakly correlated bilayers. Moreover, in the presence of salts, a weakening of van der Waals attraction forces was seen and led to a swelling of the water layer.