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Biomedical subjects

A Nicolas

Publications and source records attributed to A Nicolas.

At least 19 recordsLinked to original sources
PubMed2007

hSNF5/INI1-deficient tumours and rhabdoid tumours are convergent but not fully overlapping entities.

Rhabdoid tumours (RTs) are rare but highly aggressive tumours of childhood. Their rarity and their miscellaneous locations make the diagnosis particularly challenging for pathologists. Central nervous system and peripheral RTs have been associated with biallelic inactivation of the hSNF5/INI1/SMARCB1 (hSNF5/INI1) tumour suppressor gene. Immunohistochemistry (IHC) with a monoclonal anti-hSNF5/INI1 antibody has recently been proposed as an efficient diagnostic tool for RTs. We have conducted a retrospective study of 55 tumours referred to our institution with a suspicion of RT. This analysis included pathological review, IHC with anti-hSNF5/INI1 antibody, and molecular investigation using quantitative DNA fluorescent analysis and sequencing of the nine exons of hSNF5/INI1. The molecular lesion could be detected in 37 of the 39 cases exhibiting negative staining for hSNF5/INI1. In the two discrepant cases, the lack of detection of genetic abnormality was probably owing to the presence of a high number of non-tumour cells in the samples. This indicates that hSNF5/INI1 IHC is very sensitive and highly specific for the detection of hSNF5/INI1 loss-of-function. Among the 38 cases with typical RT histological features, six failed to exhibit hSNF5/INI1 mutation and stained positive for hSNF5/INI1. This strongly supports the evidence of a second genetic locus, distinct from hSNF5/INI1, associated with RT. Conversely, seven tumours with histological features poorly compatible with RT stained negative for hSNF5/INI1; they nevertheless exhibited an age of onset and a clinical behaviour similar to RT. This suggests that hSNF5/INI1 inactivation is not strictly limited to typical RT but characterizes a wider family of hSNF5/INI1-deficient tumours. Consequently, we believe that anti-hSNF5/INI1 IHC should be performed widely, even when the pathological characteristics are not typical. The molecular investigation should be performed in infants when a rhabdoid predisposition syndrome is suspected.

Adult↗
PubMed2006

The influence of circadian rhythm during a sustained submaximal exercise and on recovery process.

The purpose of this study was to examine the time-of-day effects on muscle fatigue and recovery process following an isometric fatiguing contraction. Sixteen male subjects were tested at two times (06:00h and 18:00h) and were requested to perform a sustained submaximal contraction of the elbow flexors, consisting in maintaining 40% of their absolute strength as long as they could. Isometric maximal voluntary contractions (MVC) were performed before (Pre), immediately after (Post), and up to 10min after the endurance task. Endurance time, peak torque (PT) and electromyographic (EMG) activities of the biceps brachii and triceps brachii were recorded and analysed. Results showed that under Pre-test conditions, PT developed at 18:00h was higher than at 06:00h. No time-of-day effect appears for the endurance time and EMG activities during the test. No time-of-day effect was observed on either MVC or EMG recovery. From the results of this study, it seems that both muscle fatigue and recovery process are not time-of-day dependent. We conclude that circadian rhythm of the force do not influence the evaluation of muscle capacities during a submaximal exercise corresponding at 40% of MVC.

Adult↗
PubMed2006

The effect of pedal rate and time of day on the time to exhaustion from high-intensity exercise.

The aim of this study was to examine the supposed influence of pedal rate on the diurnal fluctuation of the time to exhaustion from high-intensity exercise. Eleven male cyclists performed three tests at 06:00 h and three at 18:00 h at a free pedal rate (FPR) and two imposed pedal rates (80% and 120% of the FPR). They performed the tests until exhaustion using a power output corresponding to 95% maximal power (Pmax). Time to exhaustion, rectal temperature, oxygen consumption (.VO2), M. quadriceps, vastus medialis, M. biceps femoris electromyographic Root Mean Square activity rise (RMS slope), and blood lactate concentration were measured. The mean time to exhaustion recorded at 18:00 h (270.6+/-104.8 sec) was greater than at 06:00 h (233.9+/-84.9 sec). The time to exhaustion was significantly greater when the pedal rate was imposed at 80% versus 120% FPR. The blood lactate concentration and absolute core temperature at the point of exhaustion were significantly higher during tests done at 18:00 h. There was no diurnal variation in core temperature increase, .VO2, and RMS slope. The time-of-day effect for every variable did not depend on pedal rate. Diurnal variations in maximal aerobic endurance cannot be explained by a change in aerobic metabolism or in muscular fatigue. The origin of the diurnal variation in the time to exhaustion is likely to lie in greater participation in anaerobic metabolism. Also, the influence of temperature on neuromuscular functioning as an explanation for the diurnal variation in performance cannot be excluded in this study. The hypothesis on the basis of which pedal rate would influence diurnal variations in time to exhaustion in cycling was not validated by this research.

Adult↗
PubMed2005

Analysis of neomycin sulfate and framycetin sulfate by high-performance liquid chromatography using evaporative light scattering detection.

A rapid and simple method for the determination of main components and related substances of both neomycin sulfate and framycetin sulfate by HPLC and evaporative light scattering detection (ELSD) is described. The method was also used to determine the neomycin B and the sample sulfate content. Detection and quantitation of aminoglycoside antibiotics are problematic because of the lack of UV absorbing chromophore. The use of a universal detector avoids the need for sample derivatization or use of specific detector based on pulsed amperometry described to be difficult in routine assays. Separation was performed with a Polaris C18 150 mm x 4.6 mm i.d., 3 microm reversed-phase column with a solution of 170mM trifluoroacetic acid (TFA) mobile phase at a flow rate of 0.2 mL/min. The chromatographic parameters were optimized with the help of experimental design software. Mass spectrometry (MS) was employed to confirm the ELSD profile. The final method was validated using methodology described by the International Conference of Harmonization in the field of Active Pharmaceutical Ingredients. Commercial samples of different sources were analyzed and results were in good agreement with specifications of the European Pharmacopoeia.

Anti-Bacterial Agents↗
PubMed2005

Time-of-day effects on myoelectric and mechanical properties of muscle during maximal and prolonged isokinetic exercise.

The aim of this study was to examine the time-of-day (TOD) effects in myoelectric and mechanical properties of muscle during a maximal and prolonged isokinetic exercise. Twelve male subjects were asked to perform 50 maximal voluntary contractions (MVC) of the knee extensor muscles at a constant angular velocity of 2.09 rad . sec(-1), at 06 : 00 and 18 : 00 h. Torque and electromyographic (EMG) parameters were recorded for each contraction, and the ratio between these values was calculated to evaluate variations of the neuromuscular efficiency (NME) with fatigue and with TOD. The results indicated that maximal torque values (T(45)Max) was significantly higher (7.73%) in the evening than in the morning (p<0.003). The diurnal variation in torque decrease was used to define two phases. During the first phase (1st to the 26th repetition), torque values decreased fast and values were higher in the evening than in the morning, and during the second phase (27th to the 50th repetition), torque decreased slightly and reached a floor value that appeared constant with TOD. The EMG parameters (Root Mean Square; RMS) were modified with fatigue, but were not TOD dependent. The NME decrease-significantly with fatigue, showing that peripheral factors were mainly involved in the torque decrease. Furthermore, NME decrease was greater at 18 : 00 than at 06 : 00 h for the vastus medialis (p<0.05) and the vastus lateralis muscles (p<0.002), and this occurred during the first fatigue phase of the exercise. In conclusion, the diurnal variation of the muscle fatigue observed during a maximal and prolonged isokinetic exercise seems to reflect on the muscle, with a greater contractile capacity but a higher fatigability in the evening compared to the morning.

Adult↗
PubMed2004

Elastic deformations of grafted layers with surface stress.

We calculate the elastic response of thin films grafted to a solid substrate whose upper surface is subject to a stress. This issue is addressed in the context of biological cell adhesion where adhesive junctions consist of a thin layer of proteins grafted to the extracellular matrix and sheared by the cell contractility apparatus. We show that the finite thickness of the layer limits stress-induced deformations to short ranges proportional to the thickness of the film. In addition, we show that the attachment boundary condition creates an effective shear response to surface stresses that couples all the directions, even for fluidlike layers. We predict that perturbations with wavelengths of order of the film thickness induce resonancelike responses for isotropic rubberlike materials or anisotropic media with high shear moduli. We use these results to predict the elastic deformations of a layer of proteins under shear stress and propose that the resulting, polarized elastic response to local surface forces can explain the observed, anisotropic growth of cell-substrate junctions when subject to external stresses.

Algorithms↗
PubMed2004

Determination of gentamicin sulfate and related compounds by high-performance liquid chromatography with evaporative light scattering detection.

A rapid and simple method for the separation and quantitation of gentamicin sulfate by HPLC coupled with evaporative light scattering detection (ELSD) has been developed. Detection of the different components of gentamicin is problematic because of the lack of UV absorbing chromophore. The use of the universal ELSD avoids the need for sample derivatization or use of specific detector such as pulsed amperometry. Separation was performed on a highpurity C18 125 mm x 4 mm i.d., 3 microm, reversed phase column with 48.5 mM trifluoroacetic acid-methanol (97:3, v/v), as mobile phase at a flow rate of 0.7 ml/min. The influence of the gas nature, gas pressure and temperature of the drift tube of the detector on the detection response was investigated. Optimization was performed with the help of a specific experimental design software. This method allows the determination of the composition in components C1, C1a, C2, C2a and C2b of gentamicin sulfate samples. Mass spectrometry was employed to confirm the ELSD chromatographic profile. The method was validated using methodology described by the International Conference of Harmonization in the field of Medicinal Substances. Commercial samples of different sources were analyzed and results were in good agreement with specifications of both European and United States Pharmacopoeia.

Anti-Bacterial Agents↗
PubMed2004

GermOnline, a cross-species community knowledgebase on germ cell differentiation.

GermOnline provides information and microarray expression data for genes involved in mitosis and meiosis, gamete formation and germ line development across species. The database has been developed, and is being curated and updated, by life scientists in cooperation with bioinformaticists. Information is contributed through an online form using free text, images and the controlled vocabulary developed by the GeneOntology Consortium. Authors provide up to three references in support of their contribution. The database is governed by an international board of scientists to ensure a standardized data format and the highest quality of GermOnline's information content. Release 2.0 provides exclusive access to microarray expression data from Saccharomyces cerevisiae and Rattus norvegicus, as well as curated information on approximately 700 genes from various organisms. The locus report pages include links to external databases that contain relevant annotation, microarray expression and proteome data. Conversely, the Saccharomyces Genome Database (SGD), S.cerevisiae GeneDB and Swiss-Prot link to the budding yeast section of GermOnline from their respective locus pages. GermOnline, a fully operational prototype subject-oriented knowledgebase designed for community annotation and array data visualization, is accessible at http://www.germonline.org. The target audience includes researchers who work on mitotic cell division, meiosis, gametogenesis, germ line development, human reproductive health and comparative genomics.

Animals↗
PubMed2004

Increased cell size and Akt activation in HER-2/neu-overexpressing invasive ductal carcinoma of the breast.

AIMS: To determine whether cell size is related to HER-2/neu status and/or to Akt activation in breast carcinomas. HER-2/neu overexpression is observed in 20-30% of invasive breast carcinomas with poor pronostic features, but little is known about the cell phenotype associated with HER-2/neu activation. Akt has been found to be involved in the HER-2/neu signal transduction pathway and Akt activation has been associated with increased cell size in various models. METHODS AND RESULTS: A case-control study of invasive ductal carcinoma of the breast was carried out, including 21 cases displaying HER-2/neu overexpression and 20 HER-2/neu negative controls. Cytoplasmic and nuclear sizes were measured on digitized histological pictures using cell image analysis software. Akt expression analysis was performed by immunohistochemistry on formalin-fixed histological sections using an anti-phosphorylated-Akt (Ser473) antibody. RESULTS: HER-2/neu-overexpressing carcinomas had a mean nuclear size of 75 +/- 22.2 micro m(2) and a mean cytoplasmic size of 187 +/- 52.3 micro m(2). Both values were higher than the nuclear and cytoplasmic size of HER-2/neu-negative cases (nucleus = 58 +/- 24.5 micro m(2), cytoplasm = 133 +/- 56.6 micro m(2); P = 0.02 and P =0.003, respectively). Up to 75% of the tumours with a cell size over 140 micro m(2) were HER-2/neu-positive. Immunohistochemical Akt expression was observed in 19/40 (47.5%) cases. The immunoreactivity was localized in the cytoplasm in eight cases, on the cell membrane in four cases and at both sites in seven cases. One case was not interpretable. Comparison between HER-2/neu and Akt status showed that Akt was detectable at the cell membrane in 43% (9/21) of HER-2/neu-positive and in 10% (2/19) of HER-2/neu-negative cases (P = 0.02). CONCLUSIONS: HER-2/neu overexpression was consistently associated with increased cell size in invasive ductal carcinoma of the breast. This increase may be related to concomitant Akt activation. The assessment of activated pathways in HER-2/neu-overexpressing breast carcinomas may provide useful information for optimized individual HER-2/neu-targeted therapy.

Breast Neoplasms↗
PubMed2003

Interlaboratory study of a liquid chromatography method for erythromycin: determination of uncertainty.

Erythromycin is a mixture of macrolide antibiotics produced by Saccharopolyspora erythreas during fermentation. A new method for the analysis of erythromycin by liquid chromatography has previously been developed. It makes use of an Astec C18 polymeric column. After validation in one laboratory, the method was now validated in an interlaboratory study. Validation studies are commonly used to test the fitness of the analytical method prior to its use for routine quality testing. The data derived in the interlaboratory study can be used to make an uncertainty statement as well. The relationship between validation and uncertainty statement is not clear for many analysts and there is a need to show how the existing data, derived during validation, can be used in practice. Eight laboratories participated in this interlaboratory study. The set-up allowed the determination of the repeatability variance, s(2)r and the between-laboratory variance, s(2)L. Combination of s(2)r and s(2)L results in the reproducibility variance s(2)R. It has been shown how these data can be used in future by a single laboratory that wants to make an uncertainty statement concerning the same analysis.

Anti-Bacterial Agents↗
PubMed2003

Polymers grafted to a fluid and flexible membrane: extreme sensitivity to the grafting density.

Equilibrium phase coexistence between two chemical species implies the equality of the chemical potentials and of the osmotic pressures. We study this problem on a deformable membrane when one type of the molecules serves as anchor for polymeric chains immersed in the surrounding medium (considered as a good solvent). We derive the general conditions for phase coexistence when both the curvature of the membrane and the density field of the anchor molecule are free to adjust themselves. We show that curvature favors phase segregation. Our model predicts that membranes decorated with polymeric chains exhibit new shape bifurcations without equivalent in fixed density systems.

Computer Simulation↗
PubMed2002

[Human infertility: meiotic genes as potential candidates].

Up to now, the identification of gene mutations causing infertility in humans remains poorly investigated. Temporal progression through meiosis and meiosis specific genes had been extensively characterized in yeast. Recently some mammalian homologous were found. The molecular mechanisms regulating entry into and progression through meiosis in mammals are still unknown. However, disruption of some meiotic genes in mouse showed an essential role of them in meiotic chromosome synapsis and gametogenesis. Moreover, the phenotype of gonads in null mutant mice for some meiotic genes (failure to initiate or blockage in meiosis, lack of gametes or small size of gonads...) could be strikingly similar to clinical observations found in human infertility. The aim of this study was to identify putative mutations in 5 meiotic genes of several clinically well-characterized patients who present unexplained infertility (normal karyotype, women with premature ovarian failure, men with azospermia and without Y micro-deletion). For this purpose, the exons of these 5 genes (DMC1, SPO11, MSH4, MSH5, CCNA1) were all amplified by PCR with specific primers and each amplified-exon was sequenced. Sequences were aligned in comparison to the human corresponding gene available in Genbank. Many heterozygous mutations were found in different genes. Two homozygous mutations were found in MSH4 and DMC1 genes in a young man presenting a testis vanishing syndrome and a woman presenting a premature ovarian failure, respectively. Consequences of such mutations will be examined and verified in model organisms (yeast, mouse) to check the relevance of the mutations in clinical setting.

Adenosine Triphosphatases↗
PubMed2001

The binding site of acetylcholine receptor as visualized in the X-Ray structure of a complex between alpha-bungarotoxin and a mimotope peptide.

We have determined the crystal structure at 1.8 A resolution of a complex of alpha-bungarotoxin with a high affinity 13-residue peptide that is homologous to the binding region of the alpha subunit of acetylcholine receptor. The peptide fits snugly to the toxin and adopts a beta hairpin conformation. The structures of the bound peptide and the homologous loop of acetylcholine binding protein, a soluble analog of the extracellular domain of acetylcholine receptor, are remarkably similar. Their superposition indicates that the toxin wraps around the receptor binding site loop, and in addition, binds tightly at the interface of two of the receptor subunits where it inserts a finger into the ligand binding site, thus blocking access to the acetylcholine binding site and explaining its strong antagonistic activity.

Amino Acid Sequence↗
PubMed2001

Histochemical method for characterization of enzyme crystals: application to crystals of Torpedo californica acetylcholinesterase.

Histochemical methods are employed to detect and localize a wide range of enzymes. Even though protein crystallographers do not commonly use this technique, the extensively used colorimetric reaction of Karnovsky was successfully adapted for easy and quick identification of acetylcholinesterase crystals. The method relies on the reduction of ferricyanide to ferrocyanide by thiocholine, released from acetylthiocholine by enzymatic hydrolysis, followed by formation of a cupric ferrocyanide precipitate, and allows rapid differentiation between salt and enzyme crystals and between native and inhibited crystals of the enzyme.

Acetylcholinesterase↗
PubMed2001

Links between replication and recombination in Saccharomyces cerevisiae: a hypersensitive requirement for homologous recombination in the absence of Rad27 activity.

The RAD27 gene of Saccharomyces cerevisiae encodes a 5'-3' flap exo/endonuclease, which plays an important role during DNA replication for Okazaki fragment maturation. Genetic studies have shown that RAD27 is not essential for growth, although rad27 Delta mutants are temperature sensitive. Moreover, they exhibit increased sensitivity to alkylating agents, enhanced spontaneous recombination, and repetitive DNA instability. The conditional lethality conferred by the rad27 Delta mutation indicates that other nuclease(s) can compensate for the absence of Rad27. Indeed, biochemical and genetical analyses indicate that Okazaki fragment processing can be assured by other enzymatic activities or by alternative pathways such as homologous recombination. Here we present the results of a screen that makes use of a synthetic lethality assay to identify functions required for the survival of rad27 Delta strains. Altogether, we confirm that all genes of the Rad52 recombinational repair pathway are required for the survival of rad27 Delta strains at both permissive (23 degrees C) and semipermissive (30 degrees C) temperatures for growth. We also find that several point mutations that confer weaker phenotypes in mitotic than in meiotic cells (rad50S, mre11s) and additional gene deletions (com1/sae2, srs2) exhibit synthetic lethality with rad27 Delta and that rad59 Delta exhibits synergistic effects with rad27 Delta. This and previous studies indicate that homologous recombination is the primary, but not only, pathway that functions to bypass the replication defects that arise in the absence of the Rad27 protein.

Alleles↗
PubMed2001

Long term outcome of small size invasive breast carcinomas independent from angiogenesis in a series of 685 cases.

BACKGROUND: To document the role of neoangiogenesis in the progression of breast carcinomas, intratumoral vascular density (ITVD) was assessed and compared to pathologic data and disease outcome in a series of 685 cases. METHODS: Patients were registered between 1981 and 1988 at the Curie Institute. Tumors corresponded to small size (< or = 30 mm) invasive carcinomas, 71% of which were axillary lymph node-negative. In all cases, conservative surgery was the initial therapeutic procedure. The median follow-up was 10.8 years. ITVD was retrospectively determined as the number of immunostained (anti-F8RA/vWF antibody) vessels in an area of 1.2 mm(2). The prognostic value of ITVD regarding overall survival, locoregional recurrence-free, and metastasis-free intervals was assessed in uni- and multivariate analyses. RESULTS: Microvessel count ranged from 5--245 per 1.2 mm(2) field. The median value was 62, and the mean was 67. The median was chosen as a cut point for statistical analysis. ITVD was found to be inversely linked to tumor size (P < or = 0.0001) and histologic grade (P = 0.005), and directly linked to vascular invasion (P = 0.02). In uni- and multivariate analysis, no significant link was found between ITVD and disease outcome, even after adjustment on histologic grade and tumor size. CONCLUSIONS: ITVD was inversely correlated to tumor size and histologic grade in our series of small-size breast carcinomas. No significant link between ITVD and disease outcome was observed. Evaluation of the role of angiogenesis in tumor progression should be based on the discriminative assessment of mature and/or activated vessels.

Adult↗