Search PubMed⌕ Search

Biomedical subjects

M Picard

Publications and source records attributed to M Picard.

At least 55 records · Page 3Linked to original sources
PubMed2000

Biological activities of recombinant chicken leptin C4S analog compared with unmodified leptins.

The chicken leptin sequence, in contrast to mammalian leptins, contains an unpaired Cys at position 3 of the original cDNA (AF012727). The presence of an extra Cys may confer a different structure and affect the leptin's biological activity. To address this, we studied the effects of wild-type and mutated (C4S) chicken leptins in vitro and in vivo and compared them with mammalian leptin prepared from ovine leptin cDNA. The prokaryotic expression vector pMON, encoding full-size A(-1) chicken leptin (AF012727), was mutated using a mutagenesis kit, yielding the C4S analog. Escherichia coli cells transformed with this vector overexpressed large amounts of chicken leptin C4S upon induction with nalidixic acid. The expressed protein, found in the inclusion bodies, was refolded and purified to homogeneity on a Q-Sepharose column, yielding three electrophoretically pure fractions, eluted from the column by 100, 125, and 150 mM NaCl, respectively. All three fractions showed a single band of the expected molecular mass (16 kDa) and were composed of >95% monomeric protein. Proper refolding was evidenced by comparing the circular dichroism spectrum of the analog with spectra of nonmutated chicken and ovine leptins. The biological activity of the C4S analog was evidenced by its ability to stimulate proliferation of leptin-sensitive BAF/3 cells transfected with a long form of human leptin receptor construct similar to its nonmutated counterpart, indicating that Cys4 plays no role in leptin activity. The in vitro activity of both wild-type and mutated chicken leptins was approximately 10-fold lower than that of ovine leptin. After intravenous or intraperitoneal injections, C4S analog and the nonmutated chicken and ovine leptins all lowered the food intake of starved 9-day-old broiler or 5-wk-old layer male chickens by 11-34%. Monitoring food behavior revealed that the attenuated food intake resulted not from a decreased number of approaches to the feeders but from a decrease in the average time spent eating during each approach.

Aging↗
PubMed2000

Subacute but not acute generation of nitric oxide in focal cerebral ischemia.

BACKGROUND AND PURPOSE: Excessive release of nitric oxide (NO) has been implicated in the pathophysiology of neurodegeneration in ischemic stroke. We compared intracerebral release of indicators of NO generation at the acute and subacute stages of transient focal cerebral ischemia. METHODS: In vivo microdialysis in the rat striatum was performed at the acute (first hours) and subacute (after 24 or 48 hours) stages of cerebral ischemia or sham operation to monitor intracerebral release of the stable NO metabolites nitrite and nitrate. RESULTS: Whereas only a nonsignificant trend toward increased release of these NO metabolites was evidenced in acute cerebral ischemia, a significant NO generation was observed subacutely, 48 hours after induction of cerebral ischemia. Aminoguanidine, a selective inhibitor of inducible NO synthase, suppressed this delayed release of nitrite and nitrate. CONCLUSIONS: Whereas these observations do not support a major NO generation in acute cerebral ischemia, they indicate an inducible NO synthase-dependent NO generation predominantly at the subacute phase of ischemic neurodegeneration. Therefore, NO generation may play a pathophysiological role in delayed ischemic neurodegeneration.

Animals↗
PubMed1999

Oral immunisation of swine with a classical swine fever vaccine (Chinese strain) and transmission studies in rabbits and sheep.

Seven experiments including a total of 47 pigs, 11 wild boars, 26 rabbits, 10 hares and 16 sheep were carried out to assess the efficacy, safety and transmission of the Chinese vaccine strain of the classical swine fever virus (CSFV) administrated by the oral route. Within 3 weeks after oral vaccination, a clear seroconversion occurred in the pigs. Six weeks after vaccination, vaccinated pigs were fully protected against a virulent challenge. The C-strain was not isolated from tonsils, spleen, lymph nodes, thymus, saliva, urine and faeces of pigs within 4 days after oral vaccination. In one experiment, susceptible pigs were placed in direct contact with vaccinated pigs. None of these contact-exposed pigs became serologically positive for CSFV antibodies. It is concluded that the C-strain induces protection in pigs when administrated by the oral route and is not shed by vaccinated pigs. Serum anti-CSFV antibodies developed in seven out of eight wild boars vaccinated by the oral route. No vaccine virus was detected in the spleen and tonsils of these animals. The results in wild boar were in accordance with those obtained in domestic pigs. Sheep did not show any clinical signs after oral vaccination while rabbits had moderate hyperthermia and growth retardation. No clinical response to oral immunisation in hares was detected. At the end of the experiment, no sheep had detectable serum antibodies against CSFV, whereas a few vaccinated rabbits and hares became seropositive. None of the contact-exposed rabbits and hares seroconverted. These data indicate that the C-strain is safe for sheep and as expected, moderately or not pathogenic for rabbits and hares. These efficacy and safety studies on oral vaccination with the C-strain under experimental conditions provide essential information for further studies in wild boars under experimental and field conditions, including assays with baits to control a CSF epidemic.

Administration, Oral↗
PubMed1999

A simple method for testing odor detection and discrimination in chicks.

A simple procedure for testing newly hatched chicks' responses to olfactory stimuli is described. Chicks that were hand held under a heatlamp became inactive and closed their eyes (as if asleep) within 2 min. Sleeping chicks displayed overt behavioral responses to an odorized q-tip (head shaking, beak clapping) that were not observed in awake chicks. Sleeping latencies were shorter for 1-day-old chicks that had been food deprived prior to testing than for chicks that had ad lib access to food and water. When airborne odors were presented by squeezing a soft plastic odor bottle near the beak, sleeping chicks' reactions to mint were of a greater magnitude than their responses to lavender or orange scents. Nonetheless, all three odors elicited more pronounced behavioral responses than did the water control stimulus. This method allows rapid testing of individual chicks for odor detection and discrimination.

Analysis of Variance↗
PubMed1999

Regulation of cardiac hypertrophy in vivo by the stress-activated protein kinases/c-Jun NH(2)-terminal kinases.

Cardiac hypertrophy often presages the development of heart failure. Numerous cytosolic signaling pathways have been implicated in the hypertrophic response in cardiomyocytes in culture, but their roles in the hypertrophic response to physiologically relevant stimuli in vivo is unclear. We previously reported that adenovirus-mediated gene transfer of SEK-1(KR), a dominant inhibitory mutant of the immediate upstream activator of the stress-activated protein kinases (SAPKs), abrogates the hypertrophic response of neonatal rat cardiomyocytes to endothelin-1 in culture. We now report that gene transfer of SEK-1(KR) to the adult rat heart blocks SAPK activation by pressure overload, demonstrating that the activity of cytosolic signaling pathways can be inhibited by gene transfer of loss-of-function mutants in vivo. Furthermore, gene transfer of SEK-1(KR) inhibited pressure overload-induced cardiac hypertrophy, as determined by echocardiography and several postmortem measures including left ventricular (LV) wall thickness, the ratio of LV weight to body weight, cardiomyocyte diameter, and inhibition of atrial natriuretic factor expression. Our data suggest that the SAPKs are critical regulators of cardiac hypertrophy in vivo, and therefore may serve as novel drug targets in the treatment of hypertrophy and heart failure.

Adenoviridae↗
PubMed1998

A homologue of the yeast SHE4 gene is essential for the transition between the syncytial and cellular stages during sexual reproduction of the fungus Podospora anserina.

The Podospora anserina cro1 gene was identified as a gene required for sexual sporulation. Crosses homozygous for the cro1-1 mutation yield fruiting bodies which produce few asci due to the formation of giant plurinucleate cells instead of dikaryotic cells after fertilization. This defect does not impair karyogamy, but meioses of the resultant polyploid nuclei are most often abortive. Cytological studies suggest that the primary defect of the mutant is its inability to form septa between the daughter nuclei after each mitosis, a step specific for normal dikaryotic cell divisions. The cro1-1 mutant would thus be unable to leave the syncytial vegetative state while abiding by the meiotic programme. cro1-1 also shows defects in ascospore germination and growth rate. GFP-tagging of the CRO1 protein reveals that it is a cytosolic protein mainly expressed at the beginning of the dikaryotic stage and at the time of ascospore maturation. The CRO1 protein exhibits significant similarity to the SHE4 protein, which is required for asymmetric mating-type switching in budding yeast cells. Thus, a gene involved in asymmetric cell divisions in a unicellular organism plays a key role at the transition between the syncytial (vegetative) state and the cellular (sexual) state in a filamentous fungus.

Amino Acid Sequence↗
PubMed1998

Escape from Premature Death Due to Nuclear Mutations in Podospora anserina: Repeal versus Respite.

Premature death has been defined as a growth stoppage linked to the accumulation of specific deletions of the mitochondrial genome (mtDNA) in Podospora anserina. This occurs only in strains carrying the AS1-4 mutation which lies in a gene encoding a cytosolic ribosomal protein. Here we describe the isolation and genetic characterization of 10 nuclear mutations which either delay the appearance of this syndrome (respite from premature death) or cause a switch to the classical senescence process (repeal of premature death). These mutations lie in at least six genes. Some cause defects at the levels of ascospore germination, growth rates, and/or sensitivity toward inhibitors of protein syntheses. All modify the onset of senescence in wild-type (AS1+) strains. The role played by these genes is discussed with respect to the control of diseases due to mtDNA rearrangements in filamentous fungi. Copyright 1998 Academic Press.

Journal Article↗
PubMed1998

Self-selection of dietary protein and energy by broilers grown under a tropical climate: adaptation when exposed to choice feeding at different ages.

Three hundred broiler chicks (Hubbard) of both sexes were assigned to five feeding treatments: T0 received a complete diet (control), T1 was a choice feeding system with simultaneous access to an energy-rich feed (ground corn) and a protein concentrate (43.7% CP), and T2, T3, and T4 were introduced to choice feeding after 1, 3, and 5 wk, respectively, of consuming the complete diet. At 1-d-old, T1 chicks showed a marked preference for corn, with only 21.4% of their intake being the protein concentrate diet. Intake of the concentrate progressively increased to 40 to 45% after 3 d of adaptation, resulting in diets with 22.5 to 24.3% CP. When broilers fed the complete diet for 1 to 5 wk (T2, T3, T4) were changed to choice feeding, on the 1st d, their feed choice intake was similar to that of T1 chicks at the same age. Visual observation and tactile assessment of the feed particles during the initial period allowed the chicks to quickly evaluate the new feeds and to adapt their feeding behavior. Although total feed intake to 6 wk of age was not significantly affected by the feeding treatments, broilers on choice feeding selected diets with a lower percentage of CP and had lower live body weights at 6 wk than those fed the complete diet (T0).

Adaptation, Physiological↗
PubMed1997

What is a bona fide mating-type gene? Internuclear complementation of mat mutants in Podospora anserina.

In the heterothallic ascomycete Podospora anserina, the mating-type locus is occupied by two mutually exclusive sequences termed mat+ and mat-. The mat+ sequence contains only one gene, FPR1, while the mat- sequence contains three genes: FMR1, SMR1 and SMR2. Previous studies have demonstrated that FPR1 and FMR1 are required for fertilization. Further analyses have led to the hypothesis that mat+ and mat- genes establish a mat+ and mat- nuclear identity, allowing recognition between nuclei of opposite mating type within the syncytial cells formed after fertilization. This hypothesis was based on the phenotypes of strains bearing mutations in ectopic mat genes. Here we present an analysis of mutations in resident mat- genes which suggests that, unlike FMR1 and SMR2, SMR1 is not involved in establishing nuclear identity. In fact, mutations in these two genes impair nuclear recognition, leading to uniparental progeny, while mutations in SMR1 block the sexual process, probably at a step after nuclear recognition. The nuclear identity hypothesis has also been tested through internuclear complementation tests. In these experiments, the mat- mutants were crossed with a mat+ strain carrying the wild-type mat- genes. Our rationale was that internuclear complementation should not be possible for nuclear identity genes: the relevant genes should show nucleus-restricted expression, and diffusion of their products to other nuclei should not occur. This test confirmed that SMR1 is not a bona fide mat gene since it can fulfill its function whatever its location, in either a mat- or a mat+ nucleus, and even when present in both nuclei. SMR2, but not FMR1, behaves like a nuclear identity gene with respect to internuclear complementation tests. A model is proposed that tentatively explains the ambiguous behaviour of the FMR1 gene and clarifies the respective functions of the three mat- proteins.

Alleles↗
PubMed1997

Feed pecking in young chickens: new techniques of evaluation.

Three techniques were compared: automated recording (A) of 2 h of feeding activities conveyed to a computer by constantly connected electronic balances, videotaping (V) of a closeup of the head of a chick during a feed-pecking session analyzed by focal sampling at reduced speed (16 times slower), strength of pecking (S) at feed particles recorded from a feeder-weight signal conveyed to a computer by a customized electronic balance at rapid speed (24 times/s). These techniques were applied to 16-18-day-old chicks fed either a complete feed or a split diet (whole grain wheat + a complementary feed). The two feeds had similar pellet forms. The complementary feed particles were eaten at a slower rate than the complete feed particles (A and V techniques). Wheat grains were pecked with a weaker measured strength than the pellets (technique S). Two pecks of three did not result in prehension of a feed particle and were categorized as "exploratory" pecks. For 75% of the time during a continuous pecking session the head of the chick was in a static position, suggesting a long period of observation of the feed between 2 consecutive pecks. Videotaping with slow-motion focal sampling (V) offers potential development for the study of food intake behavior of chickens.

Animals↗
PubMed1997

Evaluation of gastric emptying function in clinical practice.

In this retrospective analysis, we compared different methods to evaluate gastric emptying function, aiming to improve the sensitivity and the clinical availability of our diagnostic testing. In the first study, we compared, in 72 patients clinically suspected of gastroparesis, the emptying of a meal containing two solid nutrients with different disintegration rates: 111In-labeled scrambled eggs and 99Tc-labeled liver cubes. Gastric emptying of 111In-labeled egg was delayed in 12 of our patients and the evacuation of the 99Tc-labeled liver was prolonged in 19 patients. The choice of the nutrient was not important for the identification of diabetic gastroparesis (43% vs 57%; NS), but it was determinant in the case of patients suspected of idiopathic gastroparesis (12% were positive with the egg and 25% with the liver; P < 0.05). In the second study, we compared two different diagnostic methods in 46 patients: a simple radiological detection of the gastric emptying of radiopaque pellets, and the scintigraphic emptying of a solid meal containing 99Tc-labeled liver cubes. Both tests correlated perfectly in 78% of our patients. In 15% of the population (six of these seven patients were diabetics suspected of gastroparesis) the scintigraphic method was normal, while the evacuation of radiopaque pellets was delayed. For clinical purposes, we therefore propose: (1) the scintigraphic method should use liver rather than egg as a radiolabeled tracer in order to improve the sensitivity of the test for detection of gastroparesis; and (2) the radiological detection of radiopaque markers is a reliable and convenient method for the detection of gastroparesis in clinical practice. It is possibly more sensitive than scintigraphy.

Adult↗
PubMed1997

Cholecystokinin receptors mediate the development of a preference for the mother by newly born lambs.

The aim of this study was to investigate the effect of selective cholecystokinin (CCK) antagonists on the development of a preference for the mother by newly born lambs. At birth lambs received an injection of the CCK-A antagonist devazepide (0.01 or 0.1 mg/kg), the CCK-B antagonist PD135158 (0.01 or 0.1 mg/kg), or saline for the controls (1 ml/kg). No major side effects were observed in the first 4 postnatal hours except that lambs receiving PD135158 displayed more exploratory behavior towards the maternal body than lambs from the other groups. When tested in a 2-choice test situation at 24 hr of age, lambs treated with PD135158 or saline spent significantly more time near their dams than near the alien ewes, whereas lambs treated with devazepide did not show any discrimination. The effect of devazepide persisted at 48 hr of age. The use of a CCK-A antagonist, but not a CCK-B antagonist, was concluded to prevent the formation of a preferential relationship between the lamb and its mother, most probably by impairing neonatal learning.

Animals↗
PubMed1997

Diet-induced thermogenesis in cockerels is modulated by genetic selection for high or low residual feed intake.

Energy balance of adult cockerels genetically selected for high (R+) or low (R-) residual feed consumption was investigated by using indirect calorimetry. Although no between-line difference was found in digestion of ingested energy, the true metabolizable energy (ME) intake was 40% greater in R+ than in R- birds. Basal heat production (HP) did not significantly differ between genotypes. Diet-induced thermogenesis (DIT) was significantly enhanced in R+ compared with R- birds, i.e., +84% when expressed as the difference in kilojoules of heat production determined in feed-deprived and fed cockerels, +31% when calculated as a percentage of ME intake (P < 0.01). The difference in DIT calculated from the regression between HP and physical activity explained 75% of the difference in HP; the remaining 25% could be explained by activity-related HP. The results cannot be explained by differences in the plasma concentration of circulating thyroid hormones: plasma thyroxine concentration did not differ between genotypes, whereas plasma triiodothyronine concentration was lower in feed-deprived R+ than in R- birds and indistinguishable in fed birds of the two lines. Heat production, however, was higher in the R+ line. Propranolol decreased HP only in the R+ line, suggesting a beta-adrenergic control of DIT at least in cockerels of this line. Plasma triglyceride concentration was lower in the R+ than in the R- line in fed cockerels, and plasma nonesterified fatty acid concentration was higher in the R+ than in the R- line in feed-deprived cockerels. These results are consistent with the leanness of the R+ compared with the R- line. The R+ and R- lines constitute an original model of diet-induced thermogenesis (DIT), a process that is under genetic control of appetite and allows R+ birds to balance all of their excessive energy intake without any adipose tissue storage.

Animals↗
PubMed1997

Self-selection of dietary protein and energy by broilers grown under a tropical climate: effect of feed particle size on the feed choice.

Broilers, 2 wk of age, that had been previously adapted to energy: protein choice feeding, were offered corn (either ground, cracked, or presented as whole grains) and a protein concentrate (43.7% CP) in mash or pellet form. When corn was fed as whole grains, protein concentrate in the selected diet was significantly higher (35.1%) than with cracked corn (29.3%) or ground corn (29.1%). Presenting the concentrate as pellets resulted in a significantly higher concentration in the diet (32.7%) than when mash concentrate was fed (29.6%). Live BW at 4 and 6 wk of age were not significantly affected by feed texture. However, offering corn as whole grains or concentrate as pellets induced a significant improvement in feed efficiency. Total time to eat larger size particles (whole grains, pelleted concentrate) was significantly less than total time to eat ground corn or mash concentrate. Furthermore, the mean duration of the feeding bouts was two times shorter for whole grains (48 s) than for ground corn (98 s) and for pelleted concentrate (56 s) than for mash concentrate (114 s). Chickens ate whole grains or pellets at a significantly slower rate (number of pecks per second feeding time) than when eating ground corn or mash concentrate. There was a rejection during the first 24 h when the form of the concentrate (mash to pellets) was changed. Full adaptation to the new size of the concentrate required about 3 d.

Analysis of Variance↗
PubMed1997

Responses of meat-type chickens to choice feeding of diets differing in protein and energy from hatch to market weight.

Chickens from three genetic stocks known to differ in growth potential consumed ad libitum either a single (control) diet or a choice of two diets that differed in protein and energy. Formulation of the choice diets was such that when mixed in specific proportions they provided single diets that decreased in protein and increased in energy over the experimental period. When comparisons of feeding regimens were made at a common age, body weights and feed efficiencies for all stocks were enhanced in chicks fed a single diet. When comparisons were made at a common body weight for controls, chickens fed the single diet were about 15% heavier than those given a dietary choice. For feed efficiency, however, the pattern remained for the faster growing stock whereas there was no difference between feeding regimens for the slower growing stock. Chicks provided a choice of diets had heavier abdominal fat pads and lighter breasts relative to body weight than those fed a single diet. With choice feeding, there were stock by diet interactions for dietary preferences through the first 9 d after hatch. Early on, the interactions resulted from the faster growing stocks exhibiting a greater preference for the diet higher in protein and lower in energy than the slower growing stock. By Day 5, however, the interaction occurred because stocks exhibited either no dietary preference or preferred the diet that was lower in protein and higher in energy. Regardless of genetic stock, at 9 d of age and thereafter there was a clear preference for the diet lower in protein and higher in energy than the diet higher in protein and lower in energy. These data for feed intake were consistent with behavioral observations that showed a preponderance of chicks eating from the feeder containing the diet lower in protein and higher in energy. Compared to a single diet, under choice feeding, energy utilization was negatively influenced more in the faster than slower growing stocks. Protein and sulfur amino acid utilization was not affected by feeding regimen in faster growing stocks, but was enhanced under choice feeding in the slower growing stock. Although lysine utilization was enhanced by choice feeding in all stocks, the effect was greater in the slower than in the faster growing ones. These data demonstrate that although broiler diets are formulated to enhance growth and overall feed efficiency, chicks that are provided a dietary choice of protein and energy do not eat to maximize growth or feed efficiency.

Aging↗
PubMed1997

Mutations in genes encoding the mitochondrial outer membrane proteins Tom70 and Mdm10 of Podospora anserina modify the spectrum of mitochondrial DNA rearrangements associated with cellular death.

Tom70 and Mdm10 are mitochondrial outer membrane proteins. Tom70 is implicated in the import of proteins from the cytosol into the mitochondria in Saccharomyces cerevisiae and Neurospora crassa. Mdm10 is involved in the morphology and distribution of mitochondria in S. cerevisiae. Here we report on the characterization of the genes encoding these proteins in the filamentous fungus Podospora anserina. The two genes were previously genetically identified through a systematic search for nuclear suppressors of a degenerative process displayed by the AS1-4 mutant. The PaTom70 protein shows 80% identity with its N. crassa homolog. The PaMdm10 protein displays 35.9% identity with its S. cerevisiae homolog, and cytological analyses show that the PaMDM10-1 mutant exhibits giant mitochondria, as does the S. cerevisiae mdm10-1 mutant. Mutations in PaTOM70 and PaMDM10 result in the accumulation of specific deleted mitochondrial genomes during the senescence process of the fungus. The phenotypic properties of the single- and double-mutant strains suggest a functional relationship between the Tom70 and Mdm10 proteins. These data emphasize the role of the mitochondrial outer membrane in the stability of the mitochondrial genome in an obligate aerobe, probably through the import process.

Amino Acid Sequence↗