Salivary protein profiles are linked to bitter taste acceptance in infants.

UNLABELLED: Based on recent studies in adult subjects, saliva composition is increasingly considered as a physiological factor contributing to taste sensitivity or acceptance. In order to evaluate a possible link between salivary protein composition and taste acceptance in infants, 73 infants participated longitudinally in taste acceptance tests and donated saliva at the age at 3 and 6 months. Intake ratios, reflecting acceptance of a taste solution relative to water were calculated for the five basic tastes. Salivary proteins were separated by one-dimensional electrophoresis and bands were semi-quantified by image analysis. Partial least square (PLS) regression analyses were performed for each taste at both ages to explain intake ratios by band intensities. Bitterness acceptance in the younger infants was unique in the sense that salivary protein profiles could partly predict bitter taste acceptance. At that age, infants were on average indifferent to the 0.18-M urea solution, but great variability in acceptance was observed. The six bands considered as the best predictors for bitterness acceptance were identified by MALDI-TOF mass spectrometry. Higher abundance of bands containing secretory component, zinc-α-2-glycoprotein and carbonic anhydrase 6 was associated to a lower bitterness acceptance, while higher abundance of bands containing lactoperoxidase, prolactin-inducible protein and S-type cystatins was associated to a higher bitterness acceptance. In a second stage, S-type cystatin abundance was measured by Western blotting in order to tentatively confirm this particular finding in an independent group of 22 infants. Although not reaching statistical significance, probably due to a relatively small sample size, it was again observed that cystatin abundance was higher in infants accepting more readily the bitter solution over water. CONCLUSION: saliva protein composition may contribute to bitter taste acceptance in the younger infants.

Surface plasmon resonance imaging in arrays coupled with mass spectrometry (SUPRA-MS): proof of concept of on-chip characterization of a potential breast cancer marker in human plasma.

Protein biomarker discovery and validation are crucial for diagnosis, prognosis, and theranostics of human pathologies; "omics" approaches bring new insights in this field. In particular, the combination of immuno-sensors in array format with mass spectrometry efficiently extends the classical immunoassay format and includes molecular characterization. Here, we coupled surface plasmon resonance imaging (SPRi) with MALDI-TOF mass spectrometry in a hyphenated technique which enables multiplexed quantification of binding by SPRi and molecular characterization of interacting partners by subsequent MS analysis. This adds specificity, because MS enables differentiation of molecules that are difficult to distinguish by use of antibodies, for example truncation variants or protein isoforms. Proof of concept was established for detection, identification, and characterization of a potential breast cancer marker, the LAG3 protein, at ~1 µg mL(-1), added to human plasma. The analytical performance of this new method, dubbed "SUPRA-MS", was established, particularly its specificity (S/N > 10) and reliability (100 % LAG3 identification with high significant mascot score >87.9). The adjusted format for rapid, collective, and automated on-chip MALDI-MS analysis is robust at the femtomole level and has numerous potential applications in proteomics.

Saliva electrophoretic protein profiles in infants: changes with age and impact of teeth eruption and diet transition.

OBJECTIVE: The objective of this study was to describe the changes in salivary protein profiles in infants between the ages of 3 and 6 months, and to evaluate the impact of teeth eruption and introduction of solid foods on such profiles. DESIGN: 73 infants were followed longitudinally at 3 and 6 months of age. Their whole saliva proteins were separated by SDS-PAGE electrophoresis and semi-quantified by image analysis. Amylase activity was also measured on a sub-sample of the population (n=42 infants). Bands which abundance was significantly different between the two ages according to paired comparisons were identified by mass spectrometry techniques. RESULTS: Out of 21 bands, 13 were significantly different between 3 and 6 months of age. Two short variants of amylase increased in abundance with age, as did amylase activity. Other changes possibly translated developmental physiological events, for example maturation of the adaptive immune system. The balance between S-type cystatins and cystatins A and B was modified, in favour of S-type cystatins at 6 months of age. Teeth eruption resulted in an increase in albumin abundance, whilst introduction of solid foods was associated with higher levels of ß-2 microglobulin and S-type cystatins. CONCLUSIONS: Salivary profiles were modified substantially between the ages of 3 and 6 months. Both teeth eruption and diet had an impact on abundance changes for some proteins, revealing dynamic interactions between saliva proteome, oral physiology and diet.

Proteomic mapping of bezafibrate-treated human hepatocytes in primary culture using two-dimensional liquid chromatography.

Peroxisome proliferators have been extensively studied in rodents and are known to induce liver tumors, whereas the effects of these compounds are not very clearly identified in humans when they are widely exposed to herbicides, plasticizers, solvents or drugs such as the lipid-lowering fibrate bezafibrate (BEZA). We assessed the effect of BEZA on human hepatocyte proteome. Hepatocyte proteins, including those membrane-associated, were successfully extracted and separated using 2D-liquid chromatography (PF2D, Beckman coulter). Proteins that were regulated by ≥ 1.5 fold compared to controls were identified by mass spectrometry (MALDI-TOF, Bruker Daltonics) and SwissProt bank search. BEZA modified the expression of proteins involved in various metabolic pathways as well as in cell homeostasis. No marker of peroxisome proliferation was obtained but surprisingly the expression of proteins involved in liver carcinogenicity was modulated. The co-treatment of cultures with N-acetylcysteine modified the set of proteins regulated by BEZA, either by a potentiation or an inhibition of the effects. Our study points out that the hepatocellular redox environment has to be taken into account when using fibrates in therapeutics.

Revisited BIA-MS combination: entire "on-a-chip" processing leading to the proteins identification at low femtomole to sub-femtomole levels.

We present the results of a study in which biomolecular interaction analysis (BIA, Biacoretrade mark 2000) was combined with mass spectrometry (MS) using entire "on-a-chip" procedure. Most BIA-MS studies included an elution step of the analyte prior MS analysis. Here, we report a low-cost approach combining Biacore analysis with homemade chips and MS in situ identification onto the chips without elution step. First experiments have been made with rat serum albumin to determine the sensitivity and validation of the concept has been obtained with an antibody/antigen couple. Our "on-a-chip" procedure allowed complete analysis by MS/MS(2) of the biochip leading to protein identifications at low femtomole to sub-femtomole levels. Using this technique, identification of protein complexes were routinely obtained giving the opportunity to the "on-a-chip" processing to complete the BIA-MS approach in the discovery and analysis of protein complexes.

Serial evaluation of left atrial dimension after cardioversion for atrial fibrillation and relation to atrial function.

The size of the left atrium is usually increased during atrial fibrillation (AF). The aim of the present study was to evaluate changes in left atrial (LA) dimension after cardioversion for AF, and the relation between LA dimension and atrial function. The initial study population included 171 consecutive patients. Patients who had spontaneous cardioversion to sinus rhythm (56 patients) were compared with patients who had random cardio-version with drugs (50 patients) or direct-current (DC) shock (50 patients). Echocardiographic evaluations included LA size and volume. LA passive and active emptying volumes were calculated, and LA function was assessed. Atrial stunning was observed in 18 patients reverted with DC shock and in 7 patients reverted with drugs. The left atrium was dilated in all patients during AF (48 +/- 5 mm). The size of the left atrium decreased after restoration of sinus rhythm in all patients with spontaneous reversion to sinus rhythm, in 73% of patients reverted with drugs, and in 50% of patients reverted with DC shock. The comparison between patients with a normal mechanical atrial function and patients with reduced atrial function showed that a higher atrial ejection force was associated with a more marked reduction in LA size after restoration of sinus rhythm. A relation between LA volumes and atrial ejection force was observed in the group of patients with depressed atrial mechanical function (r = -0.78; p <0.001). The active emptying fraction was lower, although not significantly, in this group, whereas the conduit volume was increased. Thus, a depressed atrial mechanical function after cardioversion for AF was associated with a persistence of LA dilation.

Propafenone versus procainamide for conversion of atrial fibrillation to sinus rhythm.

BACKGROUND AND HYPOTHESIS: Antiarrhythmic drugs are widely used for treatment of atrial fibrillation (AF) and restoration of sinus rhythm. This prospective, randomized, and controlled study compared the efficiency and safety of propafenone versus procainamide for the treatment of acute AF. METHODS: In all, 117 patients (55 women, 62 men, mean age 64.2 +/- 13 years, median 63 years) who presented with AF were included in the study. Exclusion criteria were signs or symptoms of heart failure on physical examination, recent myocardial infarction or cardiac surgery, cardiogenic shock, or hypotension. Forty-one patients spontaneously recovered sinus rhythm; the remaining 76 patients were randomized to receive propafenone or procainamide. Propafenone was given at a dose of 2 mg/kg body weight intravenously (i.v.) over 30 min. Patients randomized to receive procainamide received a bolus of 100 mg i.v. administered every 5 min up to a maximum dose of 1 g. The clinical characteristics of the two groups were comparable. RESULTS: The number of patients who recovered sinus rhythm after the treatment was larger in the procainamide-treated group (Group 1) (69.5%) than in the propafenone-treated group (Group 2) (48.7%); p < 0.05. The time required for cardioversion was significantly lower in Group 1 (mean 4.1 +/- 1.3 h), than in Group 2 patients (mean 7.3 +/- 2.6 h) (p < 0.01). CONCLUSION: In the present study, procainamide was more effective than propafenone for the treatment of AF of short duration.