Yves Claude Guillaume

University of Franche-Comté, Becoinson, Franche-Comté, France

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Publications (144)349.56 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: The tetramer destabilization of transthyretin into monomers and its fibrillation are phenomena leading to amyloid deposition. Heparan sulfate proteoglycan (HSPG) has been found in all amyloid deposits. A chromatographic approach was developed to compare binding parameters between wild-type transthyretin (wtTTR) and an amyloidogenic transthyretin (sTTR). Results showed a greater affinity of sTTR for HSPG at pH 7.4 compared with wtTTR owing to the monomeric form of sTTR. Analysis of the thermodynamic parameters showed that van der Waals interactions were involved at the complex interface for both transthyretin forms. For sTTR, results from the plot representing the number of protons exchanged vs pH showed that the binding mechanism was pH-dependent with a critical value at a pH 6.5. This observation was due to the protonation of a histidine residue as an imidazolium cation, which was not accessible when TTR was in its tetrameric structure. At pH >6.5, dehydration at the binding interface and several contacts between nonpolar groups of sTTR and HSPG were also coupled to binding for an optimal hydrogen-bond network. At pH <6.5, the protonation of the His residue from sTTR monomer when pH decreased broke the hydrogen-bond network, leading to its destabilization and thus producing slight conformational changes in the sTTR monomer structure. Copyright © 2014 John Wiley & Sons, Ltd.
    Biomedical Chromatography 09/2014; · 1.95 Impact Factor
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    ABSTRACT: Patients suffering of Alzheimer's disease (AD) are characterized by a low transthyretin (TTR) level in the brain. The effect of pH and TTR concentration in the medium on the β-amyloid protein (Aβ)/heparan sulfate proteoglycan (HSPG) association mechanism were studied using a biochromatographic approach. For this purpose, HSPG was immobilized via amino groups onto the amino propyl silica pre-packed column, activated with glutaraldehyde, by using the Schiff base method. Using an equilibrium perturbation method, it was clearly shown that Aβ can be bound with HSPG. This approach allowed the determination of the thermodynamic data of this binding mechanism. The role of the pH was also analyzed. Results from enthalpy-entropy compensation and the plot of the number of protons exchanged versus pH showed that the binding mechanism was dependent on pH with a critical value at pH=6.5. This value agreed with a histidine protonation as an imidazolium cation. Moreover, the corresponding thermodynamical data showed that at pH>6.5, van der Waals and hydrogen bonds due to aromatic amino acids as tyrosine or phenylalanine present in the N-terminal (NT) part governed the Aβ/HSPG association. Aβ remained in its physiological structure in a random coil form (i.e. the non-amyloidogenic structure) because van der Waals interactions and hydrogen bonds were preponderant. At acidic pH (pH<6.5), ionic and hydrophobic interactions, created by histidine protonation and hydrophobic amino acids, appeared in the Aβ/HSPG binding. These hydrophobic and ionic interactions led to the conversion of the random coil form of Aβ into a β-sheet structure which was the amyloidogenic folding. When TTR was incubated with Aβ, the Aβ/HSPG association mechanism was enthalpy driven at all pH values. The affinity of Aβ for HSPG decreased when TTR concentration increased due to the complexation of Aβ with TTR. Also, the decrease of the peak area with the increase of TTR concentration demonstrated that this Aβ/TTR association led to the cleavage of Aβ full length to a smaller fragment. For acidic pH (pH<6.5), it was shown that the importance of the hydrophobic and ionic interactions decreased when TTR concentration increased. This result confirmed that Aβ was cleaved by TTR in a part containing only the NT part. Our results demonstrated clearly that TTR reversed the effect of acidic pH and thus played a protective role in AD.
    Journal of pharmaceutical and biomedical analysis. 05/2014; 97C:88-96.
  • Journal of Pharmaceutical and Biomedical Analysis. 01/2014; 97:88–96.
  • Claire André, Rania Aljhni, Lydie Lethier, Yves Claude Guillaume
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    ABSTRACT: A new carbon nanotube porous silica poroshell stationary phase was developed. The chromatographic support was coated with ultrashort single-wall carbon nanotubes (SWCNTs) in a noncovalent way. It was demonstrated that the porous amino silica surface of the 300 NH2 poroshell column stabilized with 1-methyl-2-pyrrolidinone efficiently and stably adsorbed SWCNTs onto the chromatographic support. It was shown that this novel poroshell carbon nanotube (CNT) stationary phase was very useful for the HPLC separation of a series of monoclonal antibodies (mAbs) in a short analysis time (<3 min). The high-performance liquid chromatography (HPLC) method was validated and was successfully tested for the fast quantitative and qualitative control of chemotherapeutic bags fabricated in a hospital pharmacy.
    Analytical and Bioanalytical Chemistry 12/2013; · 3.66 Impact Factor
  • Yves Claude Guillaume, Claire André
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    ABSTRACT: This work deals with the development of a carbon nanotube (CNT) monolithic column coated with a pyrenyl derivative as chiral selector. For this, a solution of pyrenyl neomycine A was pumping through a monolithic CNT column previously developed by our group. This coating was stable against the desorption for months when aqueous mobile phases were used. This column was applied to the chiral separation of underivatized amino acids. As well, ultra fast separations in the range of seconds were achieved using high flow-rates.
    Talanta 10/2013; 115:418–421. · 3.50 Impact Factor
  • Firas Ibrahim, Claire Andre, Anne Iutzeler, Yves Claude Guillaume
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    ABSTRACT: A biochromatographic system was used to study the direct effect of carbon nanoparticles (CNPs) on the acetylcholinesterase (AChE) activity. The AChE enzyme was covalently immobilized on a monolithic CIM-disk via its NH2 residues. Our results showed an increase in the AChE activity in presence of CNPs. The catalytic constant (kcat) was increased while the Michaelis constant (Km) was slightly decreased. This indicated an increase in the enzyme efficiency with increase of the substrate affinity to the active site. The thermodynamic data of the activation mechanism of the enzyme, i.e. ΔH* and ΔS*, showed no change in the substrate interaction mechanism with the anionic binding site. The increase of the enthalpy (ΔH*) and the entropy (ΔS*) with decrease in the free energy of activation (Ea) was related to structural conformation change in the active site gorge. This affected the stability of water molecules in the active site gorge and facilitated water displacement by substrate for entering to the active site of the enzyme.
    Journal of Enzyme Inhibition and Medicinal Chemistry 10/2013; 28(5):1010-4. · 1.50 Impact Factor
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    ABSTRACT: Acetylcholinesterase (AChE) is a serine protease that hydrolyzes the neurotransmitter acetylcholine. Here, the effects of hydroxyl radical (OH˙) and nitric oxide (NO) on AChE activity were studied using a biochromatographic process. The enzyme was immobilized on an ethylenediamine (EDA) monolithic convective interaction media (CIM) disk. The AChE enzymatic mechanism was demonstrated from the chromatographic peak shape. A decrease in AChE activity was observed for each concentration of NO, while OH˙ radical formation led to an increase in the rate of enzymatic catalysis. Michaelis-Menten and Lineweaver-Burk plots were obtained in the presence or absence of the free radicals and their effects on Km and Vmax were evaluated. Our results indicated classical deactivation/activation kinetics without significant influence on the rate of substrate binding. The variation in transition state energies (ΔΔGES) induced by the free radicals indicated that a conformational change was occurring in the active site, while changes in the binding site were negligible. These results clearly demonstrate the direct role of OH˙ and NO on AChE activity and confirm the role they may play in Alzheimer’s disease.
    Journal of Molecular Catalysis B Enzymatic 05/2013; · 2.82 Impact Factor
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    ABSTRACT: The influence of the mobile phase composition and column temperature on the chromatographic separation of five buckminsterfullerenes (C60, C70, C76, C78, C84) on a stationary phase based on silica gel with chemically bonded humic acid (Bonded humic acid column (BHAC)) was studied. The retention behavior of the fullerenes was measured under isocratic conditions with different mobile phase compositions, ranging from 0.05–0.70 (v/v) of toluene in cyclohexane. The column temperature was analysed in the range 35–75°C. The retention factors of the five fullerenes do not depend linearly on the toluene fraction but follow a quadratic relationship. The best chromatographic conditions for baseline separation of the five fullerenes were selected. The retention of the fullerenes on the HA stationary phase was strongly affected by temperature. Positive values of thermodynamic parameters (changes of enthalpy and entropy) were due to the abnormal solubility behaviour of fullerenes in toluene in the temperature range 35–75°C. The information obtained in this work makes this BHAC very simple to prepare and low cost, useful for fullerene research applications.
  • Claire André, Gwenaelle Lenancker, Yves Claude Guillaume
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    ABSTRACT: In this paper, an effective and simple method was used for the immobilization of single wall carbon nanotubes (SWCNTs) on a monolithic HPLC material containing 2μm macropore sizes and 13nm mesopore sizes. The chromatographic support was coated with ultra short SWCNTs in a noncovalent way to preserve the sp(2) nanotube structure and thus their physico-chemical properties. It was demonstrated that the amino-surface of the monolith stabilized with 1-methyl-2-pyrrolidinone efficiently and stably adsorbed SWCNTs onto the chromatographic support. It was shown that this novel stationary phase was very useful for the HPLC isocratic mode separation of a series of small aromatic compounds in a very short analysis time. The comparison with a classical equivalent C18 monolithic column showed that the SWCNT column presented the best efficiency in similar chromatographic conditions.
    Talanta 09/2012; 99:580-5. · 3.50 Impact Factor
  • Claire André, Yves Claude Guillaume
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    ABSTRACT: In this paper, a new and effective method was described for attaching gold nanoparticles (Au-NPs) on to the surface of thiol-terminated Boron Nitride Nanotubes (BNNT) functionalized with quinuclidine-3-thiol, acting as a bridging agent. The quinuclidine-3-thiol was first grafted onto the surface of the BNNTs via strong interactions between the electron pair from the nitrogen atom of the quinuclidine structure and the electronic gap from the boron atom of the BNNT. The bare surface of Au-NPs facilitates to attach on the thiol group of the thiol-terminated BNNTs. These two nanomaterials (pristine BNNTs and Au-BNNTs) were then incorporated into a monolithic polymer. The obtained monolithic BNNT and AuBNNT stationary phases were very useful columns for the HPLC isocratic mode separation of a series of benzene and naphtalene derivatives. The retention on these two stationary phases was due to the different intermolecular interactions including the dispersion interaction (area of the delocalized π bond), the dipole-dipole interactions, and the electrostatic repulsion. The presence of Au-NPs on the BNNT surface improved significantly the retention and column efficiency for compounds with thiol groups in their structure. As well, it was shown that both retention and column efficiency linearly increased with the nanotube (NT) amount in the polymerization mixture. This manuscript thus established for the first time the fact that BNNT was a very useful nanomaterial for the development of novel HPLC stationary phases and increased the performance of classical equivalent C18 monolithic columns.
    Talanta 05/2012; 93:274-8. · 3.50 Impact Factor
  • R Aljhni, F Ibrahim, Y C Guillaume, C Andre
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    ABSTRACT: The interaction of a series of steroid hormones (DHEA, progesterone, testosterone, estradiol) with serum albumin immobilized on porous silica particles and the effect of reactive oxygen species and nitric oxide on this interaction were studied using a biochromatographic approach. The determination of enthalpy and entropy changes of this binding indicated that van der Waals interactions and hydrogen bonds predominated the hormone association with albumin. Reactive oxygen species (H(2)O(2) and OH*) increased the hormone binding affinity to albumin. On the other hand, this binding was decreased with the presence of NO*. This variation was due to conformational changes in the binding region explained by the oxidation of some residues such as free thiol and arginine. The thermodynamic analysis showed that free radical affects the van der Waals forces and/or a hydrogen bond of the hormone binding with albumin. These results explained the role of reactive oxygen species and nitric oxide in the hormone free fraction level in the blood.
    Journal of pharmaceutical and biomedical analysis 03/2012; 62:129-34. · 2.45 Impact Factor
  • Claire André, Danai Agiovlasileti, Yves Claude Guillaume
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    ABSTRACT: Multiwalled carbon nanotubes have been entrapped in a porous monolithic chromatographic support. This support was used for the covalent immobilization of the arginase enzyme a novel target in hypertension. The effect of the nanotube (NT) amount into the monolith was analyzed. The obtained results demonstrated the ability of carbon nanotubes to increase significantly the performance of this novel bioactive support.
    Talanta 10/2011; 85(5):2703-6. · 3.50 Impact Factor
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    ABSTRACT: The retention mechanism of a series of peptides on a single-wall carbon nanotube (SWCNT) stationary phase inside an HPLC column was investigated over a wide range of mobile phase compositions. While the similar size C18 column exhibited an efficiency of 11.5 μm, the SWCNT column increased the efficiency, i.e. 7.10 μm at a flow rate of 0.8 mL/min, and significantly affected the separation quality of the peptides. The values of enthalpy (ΔH) and entropy (ΔS(*)) of transfer of the peptides from the mobile to the SWCNT stationary phase were determined. The method studied each factor, i.e. ACN fraction x in the ACN/water mixture and column temperature. The changes in retention factor, ΔH and ΔS(*) as a function of the ACN fraction in the mobile phase were examined. These variations are explained using the organization of ACN in clusters in the ACN/water mixture and on the steric and electronic forces implied in the retention process. The information obtained in this work makes this SWCNT stationary phase useful for peptide research and demonstrated the role of ACN to improve the separation quality.
    Journal of Separation Science 06/2011; 34(11):1221-7. · 2.59 Impact Factor
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    ABSTRACT: The current work describes the elaboration of a simple, sensitive and reliable β-casein modified glass beads, for the detection and quantification of its specific antibody anti β-casein. This is an elementary receptor without electronic part, developed by grafting glass bead surface with the antigenic β-casein via 3-aminopropyltriethoxy silane and then glutaraldehyde as cross-linker. The whole is realized by a classical process, called in two steps and in mild conditions where chemical protocol is optimized for β-casein use. The detection and quantification of the specific reaction antibody-receptor is carried out by the technique of the second antibody labeled with horse radish peroxidase (HRP). Our receptor can detect the β-casein antibody present in the serum at dilutions up to a factor 10(7) in strong ionic strength medium. The same antibody of the same serum and in the same conditions can be detected by ELISA test at dilutions up to a factor 10(5). The whole test, after our receptor realization, takes about 5h.
    Talanta 05/2011; 84(3):632-7. · 3.50 Impact Factor
  • Claire André, Guillaume Herlem, Tijani Gharbi, Yves Claude Guillaume
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    ABSTRACT: This work was dedicated to the development of a new micro immobilized enzyme reactor (IMER) by using an in situ procedure. Arginase was covalently immobilized on an ethylenediamine (EDA) monolithic convective interaction media (CIM) disk (12mm × 3mm i.d.) previously derivatized with glutaraldehyde. The activity of this IMER was investigated by inserting this micro-IMER in a HPLC system. The effect of the arginase inhibitors was evaluated by the simultaneous injection of each inhibitor with the nitro guanidino benzene (NGB) substrate. The relative IC50 values were found in agreement with those derived by the conventional spectrometric method. This arginase micro-IMER system was also used to study the effects of plant-derived products on the arginase activity. The pet ether extract from the stem bark of the plant Ficus glomerata Roxob. and the procyanidin oligomers of cocoa and chocolate inhibit the arginase activity. Our results confirmed the direct effect of some plant extracts on the arginase activity and their interest in therapies for treating several NO-dependent smooth disorders.
    Journal of pharmaceutical and biomedical analysis 04/2011; 55(1):48-53. · 2.45 Impact Factor
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    ABSTRACT: Arginase is an enzyme which plays a role in pathophysiology such as hypertension. Here we demonstrated for the first time the direct implication of pressure and OH° radical formation on the arginase activity via a novel analytical procedure. Pressure increased arginase activity in the range 12-52bars. Activation by OH° radical showed a hyperbolic response. The OH° radicals produced were significantly inhibited by sulfasalazine (SAZ) and the inhibition of OH° radicals parallels the inhibition of arginase activity.
    Journal of chromatography. B, Analytical technologies in the biomedical and life sciences 10/2010; 878(28):2826-30. · 2.78 Impact Factor
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    ABSTRACT: The anodic oxidation of concentrated glycine based aqueous electrolyte on smooth platinum electrode leads to a strongly grafted polyglycine-like coating on the surface in an irreversible way. Due to the proton affinity towards amino groups of polyglycine (PG), the electrodeposited thin film was used as receptor for solid potentiometric pH sensor. In order to reach local pH measurement, we developed miniaturized microelectrodes on glass substrate thanks to photolithography process. We used silver chloride on silver as the reference electrode. The couple (silver chloride, PG based platinum electrode) of microelectrodes gives linear potentiometric response vs. pH in the range [2-12], reversibly and with a sensitivity of 52.4 mV/pH (for 1mm electrode size). PG based pH electrode is compared to other organic polymer based pH receptor such as linear polyethylenimine (L-PEI), polyaniline (PANI) and glass membrane.
    Talanta 06/2010; 82(1):417-21. · 3.50 Impact Factor
  • Fatimata Seydou Sarr, Claire André, Yves Claude Guillaume
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    ABSTRACT: In a previous paper, using a biophysical model system to study the passive diffusion of the statin molecules through the cell membrane, our group demonstrated that statins could cross biological membrane by passive diffusion (Sarr et al. [40]). However, in the liver, the uptake of statins would also be mediated by organic anion transporting polypeptides (Oatps) like Oatp2 a member of this family. Thus, a novel biochromatographic approach was developed in our laboratory to study the transmembrane transport of statins and an Oatp2 inhibitor via this carrier family. For this, the Oatp2 protein was immobilized via its amino groups on a chromatographic support using an "in situ" immobilization technique. For the first time, using this novel biochromatographic concept, the effect of magnesium chloride salt (MgCl(2)) on the pharmacomolecule-Oatp2 binding was investigated. It was shown an Mg(2+)-dependent pharmacomolecule-protein association and a potential facilitated diffusion of these pharmacomolecules into biological membrane. This association process was due to the central positive potential pore of the Oatp2. Indeed, at pH 7.4, all the pharmacomolecules studied were ionized (i.e. negatively charged) and so interact with this positive potential pore. However, an increase of the Mg(2+) concentration led a decrease of the pharmacomolecule-Oatp2 association attributed to ion pair formations between the Mg(2+) cation and molecules. Moreover, the decrease of this affinity could be explained by an ion attraction between the Cl(-) anion of the MgCl(2) salt and the positively charged pore of the protein. This novel biochromatographic column could be useful to find a specific reversible inhibitor for these transporters and so open new perspectives to be investigated.
    Journal of pharmaceutical and biomedical analysis 05/2010; 52(1):93-8. · 2.45 Impact Factor
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    Talanta 04/2010; 82:417-421. · 3.50 Impact Factor
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    ABSTRACT: Growing evidence suggests that arginase misregulation plays a key role in the pathophysiology of essential hypertension. In the present study, we investigated the potential cardiovascular therapeutic effects of a long-term treatment with an arginase inhibitor in adult spontaneously hypertensive rats (SHR) with fully developed hypertension. Treatment of 25-week-old SHR with the arginase inhibitor N(omega)-hydroxy-nor-L-arginine (nor-NOHA, 40 mg/day for 10 weeks) sustainably reduced systolic blood pressure (-30 mmHg, P < 0.05). The antihypertensive effect of nor-NOHA was associated with changes on mesenteric artery reactivity including the restoration of angiotensin-II-induced contraction and acetylcholine-induced vasodilation to the values of normotensive Wistar Kyoto rats. Both nitric oxide synthase and cyclooxygenase-dependent mechanisms account for the improvement of endothelial function afforded by the arginase inhibitor, which in addition blunted hypertension-induced endothelial arginase I overexpression in mesenteric arteries. Nor-NOHA also prevented the remodelling of aorta as measured by collagen content and media/lumen ratio, and improved the compliance of carotid artery in SHR. Cardiac fibrosis assessed by collagen content of left heart ventricle was reduced by nor-NOHA, with no significant effect on cardiac hypertrophy. Our results report that a long-term treatment with an arginase inhibitor reduced blood pressure, improved vascular function, and reduced cardiac fibrosis in SHR with fully developed hypertension. These data suggest that arginase represents a promising novel target for pharmacological intervention in essential hypertension.
    Cardiovascular Research 03/2010; 87(3):569-77. · 5.81 Impact Factor

Publication Stats

638 Citations
349.56 Total Impact Points


  • 2000–2014
    • University of Franche-Comté
      Becoinson, Franche-Comté, France
  • 2011
    • Technological Educational Institute of Thessaly
      • Department of Medical Laboratory
      Lárisa, Thessalia, Greece
  • 1997–2001
    • Laboratoire de Chimie de Coordination.
      Tolosa de Llenguadoc, Midi-Pyrénées, France
  • 1998
    • laboratory of physics and chemistry for environment and space
      Orléans, Centre, France
  • 1995
    • Laboratoire des Sciences du Climat et l'Environnement
      Gif, Île-de-France, France