Forage system is the key driver of mountain milk specificity.

The aims of this work were to determine the effect of upland origin on milk composition when comparing similar lowland and upland production system and to highlight the factors responsible for the added value of upland milk from commercial farms. Tanker milk from 55 groups of farms (264 farms in total) in France, Slovakia, and Slovenia was collected twice during the indoor season and 3 times during the outdoor season. The tanker rounds were selected in each country to be balanced according to their origin (lowland or upland) and within upland or lowland groups, according to the forage systems: corn-based or grass-based forage system. At each milk sampling, the production conditions were recorded through on-farm surveys. The milk was analyzed for gross composition, carotenoids, minerals, fatty acids, phenolic compound derivatives, volatile organic compound concentrations, and color. The milk from upland and lowland areas differed in their contents of a few constituents. Upland milk was richer in not identified (n.i.) retention time (Rt) 13,59, 4-methylpentylbenzene, 1-methyl-2-n-hexylbenzene, and ß-caryophyllene than lowland milk. These differences could be most likely attributable to the utilization of highly diversified and extensively managed semi-natural grasslands. The higher forbs content of upland pastures could be related as well to the richness in C18:3n-3, CLA cis-9,trans-11, MUFA, and PUFA we observed in upland compared with lowland milk during the outdoor season. In contrast, grazing on lowland pastures rich in grasses gave a yellower milk that was richer in ß-carotene. Out of the few compounds showing a significant effect of origin or its interaction, most of the milk constituents were unaffected by the origin at all. However, almost all milk constituents differed according to the forage system and the season, and the differences observed between seasons can be attributed to differences in the cow diet composition.

Potential of milk fatty acid composition to predict diet composition and authenticate feeding systems and altitude origin of European bulk milk.

The aims of this work were to elucidate the potential of using milk fatty acid (FA) concentration to predict cow diet composition and altitude of bulk milk collected in 10 different European countries and to authenticate cow-feeding systems and altitude of the production area using a data set of 1,248 bulk cow milk samples and associated farm records. The predictions based on FA for cow diet composition were excellent for the proportions of fresh herbage [coefficient of determination (R2)=0.81], good for hay, total herbage-derived forages, and total preserved forages (R2>0.73), intermediate for corn silage and grass silage (R2>0.62), and poor for concentrates (R2<0.51) in the cow diet. Milk samples were assigned to groups according to feeding system, level of concentrate supplementation, and altitude origin. Milk FA composition successfully authenticated cow-feeding systems dominated by a main forage (>93% of samples correctly classified), but the presence of mixed diets reduced the discrimination. Altitude prediction reliability was intermediate (R2<0.62). Milk FA composition was not able to authenticate concentrate supplementation level in the diet (<58% of samples correctly classified). Similarly, the altitude origin was not successfully authenticated by milk FA composition (<76% of samples correctly classified). The potential of milk FA composition to authenticate cow feeding was confirmed using a data set representative of the diversity of European production conditions.

Prediction of bulk milk fatty acid composition based on farming practices collected through on-farm surveys.

The aim of this study was to predict the fatty acid (FA) composition of bulk milk using data describing farming practices collected via on-farm surveys. The FA composition of 1,248 bulk cow milk samples and the related farming practices were collected from 20 experiments led in 10 different European countries at 44°N to 60°N latitude and sea level to 2,000 m altitude. Farming practice-based FA predictions [coefficient of determination (R(2)) >0.50] were good for C16:0, C17:0, saturated FA, polyunsaturated FA, and odd-chain FA, and very good (R(2) ≥0.60) for trans-11 C18:1, trans-10 + trans-11 C18:1, cis-9,trans-11 conjugated linoleic acid, total trans FA, C18:3n-3, n-6:n-3 ratio, and branched-chain FA. Fatty acids were predicted by cow diet composition and by the altitude at which milk was produced, whereas animal-related factors (i.e., lactation stage, breed, milk yield, and proportion of primiparous cows in the herd) were not significant in any of the models. Proportion of fresh herbage in the cow diet was the main predictor, with the highest effect in almost all FA models. However, models built solely on conserved forage-derived samples gave good predictions for odd-chain FA, branched-chain FA, trans-10 C18:1 and C18:3n-3 (R(2) ≥0.46, 0.54, 0.52, and 0.70, respectively). These prediction models could offer farmers a valuable tool to help improve the nutritional quality of the milk they produce.

Authentication of cow feeding and geographic origin on milk using visible and near-infrared spectroscopy.

The ability of near-infrared spectroscopy to trace cow feeding systems and farming altitude was tested on 486 bulk milk samples from France and northwestern Italy. Milks were grouped into feeding systems according to the main forage in the diet. Partial least square discriminant analysis correctly classified 95.5, 91.5, and 93.3% of pasture versus maize silage, hay, and fermented herbage feeding systems, respectively. Discrimination was slightly less successful when diets with large proportions of the nondominant forage were included in each group. Near-infrared spectroscopy correctly discriminated no-pasture from pasture milk, even with only 30% of pasture in the diet (5.4% cross-validation error), and the error stabilized when pasture exceeded 70% (2.5% error). Near-infrared spectroscopy did not reliably trace milk geographic origin when the feeding system effect was isolated from the altitude effect. These findings may be usefully exploited for the authentication of dairy products.

Highly water-soluble prodrugs of anthelmintic benzimidazole carbamates: synthesis, pharmacodynamics, and pharmacokinetics.

Highly water-soluble prodrugs 1a- g of anthelmintic benzimidazole carbamates 2a- g were synthesized. These prodrugs combine high aqueous solubility and stability with high lability in the presence of alkaline phosphatases. The veterinary utility of 1a was shown by a pharmacodynamic and pharmacokinetic study performed in swine. Comparable anthelmintic efficacy was observed with prodrug 1a or the parent fenbendazole 2a. The pharmacokinetic results showed that 2a is better absorbed when derived from 1a than when applied as such.

[Treatment of peritoneal carcinomatosis from colorectal and appendiceal cancer by extensive cytoreductive surgery combined with hyperthermic intraoperative intraperitoneal chemotherapy]. / Traitement a visée curative des carcinoses péritonéales d'origine colorectale et appendiculaire par chirurgie maximaliste et chimiothérapie hyperthermique intrapéritonéale peropératoire.

Peritoneal carcinomatosis (PC) from colorectal cancer is a dreadful situation characterized by a rapid and mortal evolution (median survival of 5 to 7 months amongst the series published in the literature). The classical treatment includes systemic chemotherapy whether or not associated with palliative surgery. Since the early nineties, locoregional treatments combining extensive cytoreductive surgery with intraperitoneal hyperthermic chemotherapy have been developed, affording some patients a cure and yealding a 5-year survival as high as 30 to 40% in some series. However, it is associated with high morbidity and mortality rates.

Acute respiratory failure from tracheopathia osteoplastica.

Tracheopathia osteoplastica is a benign cartilaginous and osseous metaplasia of the laryngo-tracheobronchial tree diagnosed more commonly in adults over 50 years of age. We report here the case of a 54-year-old man who underwent thyroidectomy for multinodular goiter. Immediately after an uneventful surgery, he developed an acute respiratory failure with radiologic picture of Adult Respiratory Distress Syndrome. Mechanical ventilation was set up again, bronchoscopy with biopsy disclosed a massive tracheobronchial haemorrhage from a tracheopathia osteoplastica. Supportive treatment was successfully provided and the patient resumed to a normal life 14 days after the operation.

Acute respiratory failure from tracheopathia osteoplastica.

Tracheopathia osteoplastica is a benign cartilaginous and osseous metaplasia of the laryngo-tracheobronchial tree diagnosed more commonly in adults over 50 years of age. We report here the case of a 54-year-old man who underwent thyroidectomy for multinodular goiter. Immediately after an uneventful surgery, he developed an acute respiratory failure with radiologic picture of Adult Respiratory Distress Syndrome. Mechanical ventilation was set up again, bronchoscopy with biopsy disclosed a massive tracheobronchial haemorrhage from a tracheopathia osteoplastica. Supportive treatment was successfully provided and the patient resumed to a normal life 14 days after the operation.

Molecularly imprinted polymers for the determination of a pharmaceutical development compound in plasma using 96-well MISPE technology.

The use of molecularly imprinted polymers (MIPs) as sorbents for the solid phase extraction (SPE) of a pharmaceutical compound in development, prior to quantitative analysis was investigated. Three MIPs were synthesised using a structural analogue as the template molecule. Each polymer was prepared with different monomers and porogens. The MIPs were then tested for their performance both in organic and aqueous environments, the final aim being to load plasma directly onto the polymers. At an early development stage, there is a limited amount of compound available. Due to this limitation, reducing the amount of template required for imprinting was investigated. A MIP capable of extracting the analyte directly from plasma was produced. The specificity of the polymer allowed the method to be validated at a lower sensitivity than a more conventional SPE assay. For the first time, MIPs were packed into 96-well blocks enabling high throughput analysis. The analytical method was fully validated for imprecision and inaccuracy down to 4 ng/ml in plasma.

5-HT receptor subtypes involved in the spinal antinociceptive effect of acetaminophen in rats.

The present study was designed to investigate which subtype of spinal 5-HT receptors were involved in acetaminophen-induced antinociception using the paw-pressure test. Propacetamol (prodrug of acetaminophen, 400 mg/kg, injected intravenously, corresponding to 200 mg/kg of acetaminophen) produced a significant antinociceptive effect in this test. This effect was at least partially inhibited by intrathecal (i.t.) pretreatment with the 5-HT(1B) (penbutolol), 5-HT(2A) (ketanserin), 5-HT(2C) (mesulergine) receptor antagonists, but not by the 5-HT(1A) (N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2-pyridinyl)cyclohexanecarboxamide trihydrochloride, WAY 100635) and 5-HT(3) (granisetron) receptor antagonists. This profile was very close to that obtained recently with 5-HT, which suggests an implication of 5-HT in the spinal antinociceptive effect of acetaminophen. These results, the lack of binding of acetaminophen to 5-HT receptors and the increase of central 5-HT levels induced by this drug suggest that acetaminophen-induced antinociception could be indirectly mediated by 5-HT.

Syntheses of 2-alkylamino- and 2-dialkylamino-4,6-diarylpyridines and 2,4,6-trisubstituted pyrimidines using solid-phase-bound chalcones.

Several substituted 2- and 4-hydroxyacetophenones are linked to Wang resin via a modified Mitsunobu protocol. These resin-bound acetophenones are condensed with aromatic aldehydes, and the resulting chalcones 5 are used for the synthesis of 2-dialkylamino- (9a-d) and 2-alkylamino-4,6-diarylpyridines (11a-f), and 2-alkyl-4,6-diaryl- (14a) and 2,4,6-triarylpyrimidines (14b,c) in a manner suitable for combinatorial applications.

Effects of quinidine, verapamil, nifedipine and ouabain on hysteresis in atrial refractoriness in the conscious dog: an approach to ionic mechanisms.

1. This work determines the effects of quinidine, verapamil, nifedipine and ouabain on the hysteresis of the atrial effective refractory period (AERP) in the conscious dog. 2. AERP was always longer in the increasing phase than in the decreasing phase of the extrastimulus method, thus demonstrating the existence of AERP hysteresis. Calculated as the difference between the two values, hysteresis was between 8+/-0.8 and 11+/-1.0 msec. 3. Quinidine increased hysteresis from 9+/-0.7 to 13+/-0.7 msec, whereas verapamil decreased it from 10+/-0.9 to 5+/-0.5 msec and nifedipine did not affect it. Ouabain also lengthened hysteresis from 8+/-0.8 to 11+/-1.2 msec. 4. Thus, these results confirm the existence of a hysteresis phenomenon in the AERP in the conscious dog and are evidence that the fast sodium and slow calcium specific membrane currents participate in this phenomenon.

Determination of reduced and oxidized homocysteine and related thiols in plasma by thiol-specific pre-column derivatization and capillary electrophoresis with laser-induced fluorescence detection.

A new sensitive and rapid capillary electrophoresis (CE) assay for measuring reduced and oxidized thiols in human plasma has been developed. To prevent oxidation of the thiols, whole blood was immediately centrifuged after collection and the plasma proteins were precipitated with perchloric acid. The reduced thiols in the supernatant were derivatized quantitatively at 25 degrees C, pH 7.5 with a fluorescent reagent, fluorescein-5-maleimide (FM). The total plasma concentration of thiols, including the fraction coupled to proteins, was assayed after an initial reduction of the disulfide linkage in plasma with dithiothreitol. The separation of FM-thiols was performed in an acetonitrile/10 mM sodium phosphate-50 mM SDS buffer [25:75 (v/v); pH 7.0] using a fused-silica capillary (57 cm x 75 microm I.D.) at 45 degrees C. A 3-mW argon-ion laser (lambda(ex) 488 nm/lambda(em) 520 nm) was employed for FM-thiol detection. With the electric field of 530 V/cm, the time needed for the separation of FM-homocysteine, FM-glutathione and FM-N-acetylcysteine was less than 8 min. The lower limit of detection was 3 microM for the total thiols and 10 nM for the reduced thiols. The method was applied to, the determination of homocysteine levels in plasma from patients with end-stage renal disease.

Determination of the antitumor agent depsipeptide in plasma by liquid chromatography on serial octadecyl stationary phases.

A high-performance liquid chromatographic assay has been developed and validated for the determination of the antitumor agent depsipeptide (FR-901228) in plasma samples from patients with advanced cancer. After the plasma proteins were precipitated with acetonitrile, the supernatant was extracted with ethylacetate. Depsipeptide was chromatographed on two serial octadecylsilica stationary phases using a mobile phase consisting of acetonitrile-potassium phosphate buffer (0.03 M, pH 3) (27:73, v/v), at a flow-rate of 2.0 ml/min and at ambient temperature. The method was linear over a 50 to 2000 ng/ml range and the intra- and inter-day coefficients of variations were less than 8%. The method was applied to the determination of the plasma concentration-time profile for 14 patients with advanced cancer receiving from 1 to 7.5 mg/m2 of depsipeptide per day as a continuous 4-h infusion.

Cardiac anticholinergic effects of procainamide and its N-acetylated metabolite: experimental pharmacological and radioligand binding studies.

1. The cardiac anticholinergic effects of procainamide (1 mg kg(-1) min(-1)) and its N-acetylated metabolite (NAPA) at equimolar dose (1.16 mg kg(-1) min(-1)) were studied using in vivo experimental pharmacological and in vitro radioligand binding studies. 2. Procainamide and NAPA progressively reduced vagal stimulation-induced bradycardia in chloralose-anaesthetized dogs. As indicated by the ED50, the vagolytic activity of NAPA is 1.5-2.0 times weaker than that of procainamide. Both drugs increased heart rate, with lowering of mean blood pressure during the second part of procainamide infusion, but not during NAPA infusion. 3. Binding studies on rat heart membranes yielded Ki values that were 1.5 times higher for NAPA than for procainamide. 4. These results show that NAPA exerts a weaker cardiac vagolytic action than procainamide, which is probably linked to a lower ability to bind to cardiac muscarinic receptors.

Expression and characterization of a lactosaminoglycan-carrying glycoprotein of Zajdela hepatoma cell surface--structural analysis of the carbohydrate moiety.

In poorly differentiated hepatoma cells, a glycoprotein carrying lactosaminoglycans is identified, and the structure of its glycan moiety is proposed. After membrane solubilization, protein fractionation by gel filtration, and electroelution, this glycoprotein (GPIII) was identified by its affinity for Datura stramonium lectin and its content in large glycopeptides. As shown by PAGE, GPIII has an apparent molecular mass of 100 kDa and is highly glycosylated (36%). It appears as an integral membrane glycoprotein. It is absent from normal hepatocytes, in that no heavy glycopeptides could be detected that bound to Datura lectin or to specific antiserum. The glycan moiety of GPIII has been analyzed according to carbohydrate composition, glycosidase treatment, affinity chromatography on immobilized pokeweed, Datura and Griffonia lectins, and by NMR and methylation analyses. The glycan is a N-linked tetraantennary lactosaminoglycan of 6.6 kDa, containing Gal, GlcNAc, Man, and NeuNAc in a 16:14:3:4 molar ratio, with an average of three repeating units/branch. Its beta-Gal residues are in the penultimate position and are linked in beta1-4 at least in four structural elements (three peripheral and one internal). It contains a very branched structure with Gal alpha1-3Gal beta1-4GlcNAc side chains linked in the C6 position to an inner Gal residue in a main branch. Alpha-Gal and NeuNAc residues [mainly NeuNAc alpha(2-3) linkage] are expressed as the nonreducing terminal groups. A possible structural model is proposed for this heterogeneous lactosaminoglycan, although no definitive structure can be established. That this lactosaminoglycan-carrying glycoprotein GPIII is not expressed in hepatocytes suggests its expression to be linked to the undifferentiated and/or malignant state of this hepatoma.

Cardiac vagal effects of rilmenidine in the dog: comparison with clonidine.

1. The cardiac vagal effects of rilmenidine (5 micrograms kg-1 min-1) and clonidine (0.5 micrograms kg-1 min-1) were studied in chloralose anaesthetized dogs. 2. Rilmenidine and clonidine progressively reduced the vagal stimulation-induced bradycardia. As indicated by the ED70, rilmenidine was about 23 times less potent than clonidine in this respect. Concomitantly, both drugs dose-relatedly decreased heart rate and mean blood pressure with potency ratios of rilmenidne to clonidine of about 1:23 and 1:12, respectively. 3. Importantly, the heart rate values observed under vagal stimulation during drug infusion never exceeded the values under basal vagal stimulation, and with both drugs large interindividual variations occurred under vagal stimulation. 4. These results show that the vagal bradycardia inhibition produced by rilmenidine and clonidine results from their true bradycardic effects and not from actual cardiac vagolytic properties.

Hysteresis in atrial refractoriness in the conscious dog: influence of stimulation parameters and control by the autonomic nervous system.

This work (a) provides evidence for hysteresis in the atrial effective refractory period (AERP) in the conscious dog; (b) studies the main stimulation parameters that may affect this phenomenon; and (c) evaluates the influence of the autonomic nervous system. AERP was measured by the extrastimulus method in the conscious dog with chronic atrioventricular block (n = 6) during the increasing and decreasing phases of an S1S2 fixed protocol. AERP was longer during the increasing phase than during the decreasing phase, thus demonstrating hysteresis, calculated as the difference between the two values. Hysteresis was greater with an S1S1 basic cycle length of 300 ms than with a basic cycle length of 400 ms, 9 +/- 0.9, and 7 +/- 0.9 ms, respectively. It was also greater with trains of six basic cycles before each extrastimulus S2 than with trains of 12 basic cycles, 9 +/- 0.9 and 7 +/- 1.0 ms, respectively. Suppression of vagal tone with atropine reduced hysteresis from 8 +/- 0.6 to 4 +/- 0.6 ms, whereas suppression of cardioaccelerator tone with propranolol increased it from 9 +/- 0.9 to 14 +/- 1.2 ms. These data were confirmed by the neostigmine-induced increase in hysteresis from 8 +/- 0.8 to 11 +/- 0.8 ms and the isoproterenol-induced decrease in hysteresis from 9 +/- 0.6 to 4 +/- 0.4 ms. Overall, these results provide evidence for a hysteresis effect in the AERP in the conscious dog that is stimulation frequency-dependent and modulated by the autonomic nervous system with permanent increase by vagal tone and decrease by cardioaccelerator tone.