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Results for “Spectroscopy, Fourier Transform Infrared”

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PubMed2006

Geotrichum candidum dominates in yeast population dynamics in Livarot, a French red-smear cheese.

The diversity and dynamics of yeast populations in four batches of Livarot cheese at three points of ripening were determined. Nine different species were identified by Fourier transform infrared spectroscopy and/or sequencing, and each batch had its own unique yeast community. A real-time PCR method was developed to quantify the four main yeast species: Debaryomyces hansenii, Geotrichum candidum, Kluyveromyces sp. and Yarrowia lipolytica. Culture and molecular approaches showed that G. candidum was the dominant yeast in Livarot cheese. When D. hansenii was added as a commercial strain, it codominated with G. candidum. Kluyveromyces lactis was present only at the start of ripening. Yarrowia lipolytica appeared primarily at the end of ripening. We propose a scheme for the roles and dynamics of the principal Livarot yeasts.

Ascomycota↗

PubMed1997

Comparison of results of various methods used to determine the extent of modification of methoxy polyethylene glycol 5000-modified bovine cupri-zinc superoxide dismutase.

The protein bovine cupri-zinc superoxide dismutase (SOD) was modified by the reaction of lysine residues with an active ester of methoxy polyethylene glycol 5000 (PEG). The extent of modification was determined by capillary zone electrophoresis, matrix-assisted laser desorption/ionization mass spectrometry, Fourier-transform infrared spectroscopy, and Raman spectroscopy and after removing PEG by alkaline hydrolysis of the linkages to SOD followed by quantification of the released PEG using gel-permeation chromatography. There was generally good agreement among the results obtained by these techniques on a typical sample of the PEG-modified protein. The results, extent of protein modification, determined by the preceding methods were compared to that found from the classical trinitrobenzene sulfonic acid (TNBS) procedure. In addition to providing alternatives to the TNBS procedure, results from the described methodologies strongly suggest that the extent of modification determined from the TNBS procedure is overestimated.

Animals↗

PubMed2004

Mineralization of decalcified bone occurs under cell culture conditions and requires bovine serum but not cells.

The purpose of this study was to develop an in vitro model system for bone matrix mineralization in the absence of cells. For this model, we utilized EDTA-decalcified new-born rat tibias with the cartilaginous ends intact, allowing us to visually determine the specificity of mineralization within the bone. Our results show that supplementation of DMEM culture medium with 10mM beta-glycerophosphate and 15% fetal bovine serum (FBS) results in non-physiological mineral percipitation in the tibia because of the generation of supraphysiological (5mM) levels of inorganic phosphate in the medium. The same medium supplemented only with inorganic phosphate to a final concentration of 2mM failed to mineralize a decalcified tibia matrix. However, additional supplementation of this medium with as little as 5% FBS resulted in mineralization of those regions of the type I collagen where mineral was found prior to decalcification, with no evidence for mineralization in the cartilage at the bone ends or in the periosteum. Analysis of the mineral by Fourier-transform infrared spectroscopy and powder X-ray diffraction shows that tibias that have been decalcified and then remineralized contain an apatitic mineral that is strikingly similar to the mineral in normal bone. Tendon, a type I collagen matrix not normally mineralized in vivo, also mineralizes when incubated in DMEM containing 2mM Pi and as little as 1.5% FBS, but not when incubated in DMEM without serum. These data indicate that serum contains a nucleator of type I collagen matrix mineralization, and that mineralization of type I collagen under cell culture conditions requires serum but not living cells.

Animals↗

PubMed2007

On the suitability of nanocrystalline ferrites as a magnetic carrier for drug delivery: functionalization, conjugation and drug release kinetics.

Superparamagnetic nickel ferrite nanoparticles functionalized with polyvinyl alcohol, polyethylene oxide and polymethacrylic acid (PMAA) polymers and subsequently conjugated with doxorubicin anti-cancer drug are studied for their use as a magnetic carrier for drug delivery. Fourier transform infrared spectroscopy enabled examination of the ability of the nanoparticles to be functionalized with polymers and conjugated with doxorubicin drug. The functionalized polymer-coated nanocrystalline nickel ferrites retain the magnetic characteristics of non-functionalized nanocrystalline nickel ferrites (superparamagnetism, absence of hysteresis, remanence and coercivity at room temperature), encouraging their application as a magnetic carrier for drug delivery. The PMAA-coated nanoferrites are demonstrated as being a potentially superior magnetically targeted drug carrier based on FTIR results and drug release kinetics in the absence and presence of an external magnetic field.

Drug Delivery Systems↗

PubMed2003

Propranolol hydrochloride-anionic polymer binding interaction.

Three different anionic polymers namely Eudragit S 100, Eudragit L 100-55 (methacrylic acid copolymers), and sodium carboxymethylcellulose (NaCMC) were used to evaluate the propranolol hydrochloride-anionic polymer interaction. The physical and chemical properties of propranolol hydrochloride and anionic polymer complex were investigated using Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC). The DSC profiles demonstrated that the characteristic peak of propranolol hydrochloride cannot be found in the heating curve of the complexes, indicating that complex is different in physicochemical properties from the physical mixture of drug-polymer. The FTIR spectra also confirmed that there is an interaction between propranolol hydrochloride and methacrylic acid copolymers. The binding of the drug to the polymers was due to the existence of preferential hydrogen bonding between the amino group of the propranolol hydrochloride and the carboxylic functions of the polymers and that pH conditions can influence this binding.

Acrylic Resins↗

PubMed2004

Stabilization of model membranes during drying by compatible solutes involved in the stress tolerance of plants and microorganisms.

Many organisms accumulate compatible solutes under environmental stress conditions. Cyanobacteria accumulate compatible solutes in response to increased external salinity, with tolerance increasing from Suc (sucrose) or trehalose to 2-O-(alpha-D-glucopyranosyl)-glycerol and glycinebetaine accumulating species. It is not clear how these different solutes influence salt tolerance. One possible explanation may be a differential ability of these solutes to stabilize membranes under stress conditions. We therefore performed drying experiments with liposomes in the presence of compatible solutes. Suc, trehalose and sorbitol protected liposomes from leakage of a soluble marker and from membrane fusion during drying and rehydration. 2-O-(alpha-D-glucopyranosyl)-glycerol was less effective and glycinebetaine showed hardly any effect. In combination with Suc, the latter two solutes showed improved protection. Lipid-phase transitions are known to contribute to solute leakage from liposomes. We determined phase transitions in dry membranes in the absence or presence of the solutes, using Fourier-transform infrared spectroscopy. The ability of the solutes to decrease the phase transition temperature corresponded closely to their ability to protect the liposomes against solute leakage. All solutes interacted with the phosphate in the lipid headgroups. The magnitude of the shift in the asymmetric P=O stretching vibration correlated closely with the lipid-phase transition temperature. This indicates that the degree of membrane protection afforded by the solutes is mainly determined by their ability to interact with the membrane lipids. However, this is not a determinant of cellular protection against salt stress, as the solutes show a reverse order when ranked with regard to protection against these stresses.

Betaine↗

PubMed2002

Spectroscopic evidence for a preferential location of lidocaine inside phospholipid bilayers.

We examined the effect of uncharged lidocaine on the structure and dynamics of egg phosphatidylcholine (EPC) membranes at pH 10.5 in order to assess the location of this local anesthetic in the bilayer. Changes in the organization of small unilamellar vesicles were monitored either by electron paramagnetic resonance (EPR)-in the spectra of doxyl derivatives of stearic acid methyl esters labeled at different positions in the acyl chain (5-, 7-, 12- and 16-MeSL)-or by fluorescence, with pyrene fatty-acid (4-, 6-, 10- and 16-Py) probes. The largest effects were observed with labels located at the upper positions of the fatty-acid acyl-chain. Dynamic information was obtained by 1H-NMR. Lidocaine protons presented shorter longitudinal relaxation times (T(1)) values due to their binding, and consequent immobilization to the membrane. In the presence of lidocaine the mobility of all glycerol protons of EPC decreased, while the choline protons revealed a higher degree of mobility, indicating a reduced participation in lipid-lipid interactions. Two-dimensional Nuclear Overhauser Effect experiments detected contacts between aromatic lidocaine protons and the phospholipid-choline methyl group. Fourier-transform infrared spectroscopy spectra revealed that lidocaine changes the access of water to the glycerol region of the bilayer. A "transient site" model for lidocaine preferential location in EPC bilayers is proposed. The model is based on the consideration that insertion of the bulky aromatic ring of the anesthetic into the glycerol backbone region causes a decrease in the mobility of that EPC region (T(1) data) and an increased mobility of the acyl chains (EPR and fluorescence data).

Deuterium Oxide↗

PubMed2006

Physicochemical and functional characterization of a biosurfactant produced by Lactococcus lactis 53.

Isolation and identification of key components of the crude biosurfactant produced by Lactococcus lactis 53 was studied. Fractionation was achieved by hydrophobic interaction chromatography which allowed the isolation of a fraction rich in glycoproteins. Molecular (by Fourier transform infrared spectroscopy) and elemental compositions (by X-ray photoelectron spectroscopy) were determined. Critical micelle concentration achieved for the isolated fraction was 14 g/l, allowing for a surface tension value of 36 mJ/m(2). Moreover, the isolated fraction, stable to pH changes between 5 and 9, was found to be an anti-adhesive and antimicrobial agent against several bacterial and yeast strains isolated from explanted voice prostheses, even at low concentrations. Further purification steps should be carefully analyzed as each purification step will increase the costs and decreases the amounts of biosurfactants recovered.

Anti-Infective Agents↗

PubMed2005

Conformational changes of fibrinogen after adsorption.

The adsorption behavior of fibrinogen to two biomedical polyurethanes and a perfluorinated polymer has been investigated. Changes in the secondary structure of adsorbed fibrinogen were monitored using attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) and sum frequency generation vibrational spectroscopy (SFG). SFG measurements were performed in the amide I range as well as in the C-H/N-H stretching range. Amide I signals from SFG demonstrate that fibrinogen has post-adsorption conformational changes that are dependent upon the polymer surface properties. For example, strong attenuation of the amide I and N-H stretching signals with increasing residence time was observed for fibrinogen adsorbed to poly(ether urethane) but not for the other two polymers. This change is not readily observed by ATR-FTIR. Differences in the observed spectral changes for fibrinogen adsorbed to each polymer are explained by different initial binding mechanisms and post-adsorption conformational changes.

Adsorption↗

PubMed2004

Solid dispersion of carbamazepine in PVP K30 by conventional solvent evaporation and supercritical methods.

This study compares the physicochemical properties of carbamazepine (CBZ) solid dispersions prepared by either a conventional solvent evaporation versus a supercritical fluid process. Solid dispersions of carbamazepine in polyvinylpyrrolidone (PVP) K30 with either Gelucire 44/14 or Vitamin E TPGS, NF (d-alpha-tocopheryl polyethylene glycol 1000 succinate) were prepared and characterized by intrinsic dissolution, differential scanning calorimetry, powder X-ray diffraction and Fourier transform infrared spectroscopy. CBZ/PVP K30 and CBZ/PVP K30/TPGS solid dispersions showed increased dissolution rate. The best intrinsic dissolution rate (IDR) was obtained for supercritically processed CBZ/PVP K30 that was four-fold higher than pure CBZ. Thermograms of various solid dispersions did not show the melting peak of CBZ, indicating that CBZ was in amorphous form inside the carrier system. This was further confirmed by X-ray diffraction studies. Infrared spectroscopic studies showed interaction between CBZ and PVP K30 in solid dispersions. The amorphous state of CBZ coupled with presence of interaction between drug and PVP K30 suggests fewer, if any, stability problems. Because the supercritical-based process produced solid dispersions with IDR better than conventional solid dispersions augmented with amphiphilic carriers, stability issues associated with lipid carriers do not apply, which, in turn, implies easier scale up under current Good Manufacturing Practice for this technique.

Calorimetry, Differential Scanning↗

PubMed2007

Formation of bone-like apatite enhanced by hydrolysis of octacalcium phosphate crystals deposited in collagen matrix.

It has been shown that granules of synthetic octacalcium phosphate (OCP) or the composites with collagen are capable of enhancing bone regeneration, accompanied by a gradual conversion from OCP to apatite with time. The present study was designed to investigate whether formation of bone-like apatite can be accelerated by OCP deposited throughout collagen matrix (OCP collagen complex, OCC) immersed in simulated body fluid (SBF). The formation of bone-like apatite has been suggested to be essential to induce osteoconductivity of various substrates. The formation of OCP in collagen solution was investigated in calcium or phosphate ions in the range between 22.5 and 142.5 mM and pH 6.26 and 8.56. X-ray diffraction, Fourier transform infrared spectroscopy, and scanning electron microscopy (SEM) showed that condition to nucleate OCP was limited to that of a solution with Ca/P 0.43 around pH 7.16 in the presence of collagen. OCP was shown to be formed throughout the collagen matrix by SEM observation. The immersion of OCC in SBF up to 10 days enhanced apatite crystal deposition, probably through OCP-apatite conversion: the apatite formation in OCC took place within only 1 day. The present study indicated that the existence of OCP deposited throughout the collagen matrix promotes bone-like apatite formation under physiological condition.

Animals↗

PubMed2004

The effect of protein internal cavities on ligand migration and binding in myoglobin.

Using Fourier Transform Infrared Spectroscopy at cryogenic temperatures, we have studied carbon monoxide (CO) migration in the interior of sperm whale myoglobin and binding to teh heme iron. The effect of protein internal cavities was examined by comparing the wild-type protein with mutants in which the cavities were blocked by bulky amino acid sidechains. After photodissociation at 3 K, CO ligands reside in the primary docking site (B) from where they can migrate to other sites (C, D) that were identified as the Xe4 and Xe1 cavities. These studies were complemented by flash photolysis experiments at room temperature, which revealed that the protein cavities enable efficient excape of ligands from the protein after dissociation from the heme iron.

Animals↗

PubMed1994

Sequence and structure of the membrane-associated peptide of glycophorin A.

Glycophorin A (GPA) has been reconstituted into dimyristoylphosphatidylcholine vesicles and digested with proteinase K to identify the membrane domain and to characterize its structure and orientation. After digestion of the inner and outer domain of GPA by protease action restricted to the aqueous phase, a protected peptide migrates on an electrophoresis gel as a 7.5-kDa dimer (His66-Ile95). The secondary structure and orientation in a lipid bilayer of the 7.5-kDa dimer have been studied by Fourier transform infrared spectroscopy. Our proteolytic and spectroscopic data are in agreement with a topological model in which the His66-Glu72 peptide adopts a beta-sheet conformation and is oriented parallel to the lipid-water interface and the Ile73-Ile95 domain is helical and oriented parallel to the lipid acyl chains, in a transmembrane configuration. Digestion of the domain protruding to the outside of the liposome generates "head-head" and "head-tail" dimers of 16 and 38 kDa, respectively. This observation is discussed in terms of the specificity of the dimer formation process.

Amino Acid Sequence↗

PubMedDate not supplied

Comparison of the in vivo behavior of polyvinylidene fluoride and polypropylene sutures used in vascular surgery.

To find a nonabsorbable suture material that is equivalent to polypropylene in ease of handling and tensile properties, and that has low thrombogenicity and tissue reactivity but improved biostability, some researchers and clinicians see merit in considering the suitability of monofilaments made from polyvinylidene fluoride. The current animal study investigated the relative biocompatibility and biostability of these two suture materials by using them to anastomose a polyester arterial prosthesis in a canine thoracoabdominal bypass model for 10 periods of implantation ranging from 4 hr to 2 years. Biocompatibility was assessed with light and scanning electron microscope examinations of the explanted sutures, and biostability of the cleaned sutures was determined by Fourier transform infrared spectroscopy and scanning electron microscope analysis. The polyvinylidene fluoride and polypropylene sutures were found to have similar handling and healing characteristics. During the first months in vivo, both types of suture experienced a temporary increase in carbonyl group absorption that coincided with the duration of the inflammatory response. After 1 and 2 years in vivo, the explanted polypropylene sutures showed visual evidence of surface stress cracking. This was not found with the explanted polyvinylidene fluoride sutures. These results suggest that polyvinylidene fluoride may be more biostable than polypropylene in the long term.

Animals↗

PubMed2006

Physico-chemical characterization of lignins from different sources for use in phenol-formaldehyde resin synthesis.

During the last decades lignin has been investigated as a promising natural alternative to petrochemicals in phenol-formaldehyde (PF) resin production, due to their structural similarity. Physico-chemical characterization of three types of lignin, namely kraft pine lignin (L1), soda-anthraquinone flax lignin (L2), and ethanol-water wild tamarind lignin (L3) has been evaluated to determine which one is the most suitable chemical structure for above purpose. Characterization has been performed using Fourier transform infrared spectroscopy (FT-IR) and proton nuclear magnetic resonance spectrometry ((1)H NMR) to analyse the chemical structure, gel permeation chromatography (GPC) for determining molecular weight (MW) and molecular weight distribution (MWD), differential scanning calorimetry (DSC) to measure the glass transition temperature and thermogravimetric analysis (TGA) to follow the thermal degradation. Both structural and thermal characteristics suggest that kraft pine lignin (L1) would be a better phenol (P) substitute in the synthesis of lignin-phenol-formaldehyde (LPF) resins, as it presents higher amounts of activated free ring positions, higher MW and higher thermal decomposition temperature.

Calorimetry, Differential Scanning↗

PubMed1993

Lysophosphatidylcholine mediates the mode of insertion of the NH2-terminal SIV fusion peptide into the lipid bilayer.

We report here on the interaction of a synthetic 12 residue peptide corresponding to the N-terminal sequence of gp32 from SIV with phospholipid bilayers. This peptide has been shown to induce lipid mixing of PC/PE/SM/Chol LUV (large unilamellar vesicles) at pH 7.4 and 37 degrees C [(1992) in: Advances in Membrane Fluidity, vol. 6, pp. 365-376, Wiley-Liss]. In the present study, this fusion process was inhibited by the addition of lysophosphatidylcholine (lysoPC) to the lipid bilayer of PC/PE/SM/Chol LUV. Fourier transform infrared spectroscopy (FTIR) reveals that the orientation of the SIV fusion peptide with respect to the lipid acyl chains depends on the presence of lysoPC in the lipid bilayer but that the peptide secondary structure and the amount of lipid-associated peptides do not depend on the lipid composition. The peptide is obliquely inserted into the lipid bilayer of vesicles without lysoPC, whereas it is oriented parallel to the lipid-water interface in the vesicles containing lysoPC. The data provide evidence that the orientation of the SIV fusion peptide depends on the lipid composition, and that this mediates its fusogenic activity.

Kinetics↗

PubMed2006

Functionally graded bioactive coatings: reproducibility and stability of the coating under cell culture conditions.

This work sought to provide a basic protocol for treatment of functionally graded bioactive glass coatings (FGC) that reliably adhere to titanium alloy (Ti6Al4V) prior to in vivo evaluation. The effect of the fabrication process on glass structure and reproducibility of the coating's properties, and the effect of cell culture conditions on the integrity of the coating were assessed. The structure of FGCs was compared to that of the as cast glass used as a top coating. X-ray diffraction (XRD) showed that the fabrication process resulted in 5.9+/-3.0 vol.% crystallization, while glass as cast was amorphous. Glass as cast and coatings behaved similarly in simulated body fluid (SBF): an amorphous layer rich in phosphate formed, and it crystallized, over 4 weeks, into apatite-like mineral (Fourier transform infrared spectroscopy (FTIR), XRD, scanning electron microscopy (SEM)). Reproducibility of the fabrication process was tested from three batches of coatings by measuring thickness and crystallinity. MC3T3-E1.4 mouse pre-osteoblast cells were cultured and induced to mineralize on FGCs, either as made or pre-conditioned in SBF. The sub-surface glass silica network in FGCs was compromised by cell culture conditions. A crystalline phosphate was formed during pre-conditioning (XRD, FTIR, and SEM). SBF-pre-conditioning stabilized the coatings. Thus incubation in SBF is recommended to produce a stable coating.

3T3 Cells↗

PubMed2006

Bionanocomposites from renewable resources: epoxidized linseed oil-polyhedral oligomeric silsesquioxanes hybrid materials.

This study is concerned with the preparation and properties of a new class of bionanocomposites from renewable resources. Epoxidized linseed oil (ELO) and 3-glycidylpropylheptaisobutyl-T8-polyhedral oligomeric silsesquioxane (G-POSS) (2, 5, and 10 wt %) were cross-linked, and Fourier transform infrared spectroscopy (FTIR), dynamic mechanical thermal analysis (DMTA), thermogravimetric analysis (TGA), and scanning electron microscopy (SEM) were employed to characterize the POSS-reinforced oil-based polymer networks. No POSS aggregates were observed for the 2 wt % G-POSS nanocomposite by SEM. POSS-rich particles with diameters of several nanometers were observed in the nanocomposites with 5 and 10 wt % G-POSS. Enhanced glass transition temperatures and storage moduli of the networks in the glassy state and rubber plateau were observed to be higher than those of the POSS-free oil based polymer network, due to the reinforcement effect of POSS cages.

Biocompatible Materials↗