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PubMed2006

Prion protein helix1 promotes aggregation but is not converted into beta-sheet.

Prion diseases are caused by the aggregation of the native alpha-helical prion protein PrP(C) into its pathological beta-sheet-rich isoform PrP(Sc). In current models of PrP(Sc), helix1 is assumed to be preferentially converted into beta-sheet during aggregation of PrP(C). This was supported by the NMR structure of PrP(C) since, in contrast to the isolated helix1, helix2 and helix3 are connected by a small loop and are additionally stabilized by an interhelical disulfide bond. However, helix1 is extremely hydrophilic and has a high helix propensity. This prompted us to investigate the role of helix1 in prion aggregation using humPrP(23-159) including helix1 (144-156) compared with the C-terminal-truncated isoform humPrP(23-144) corresponding to the pathological human stop mutations Q160Stop and Y145Stop, respectively. Most unexpectedly, humPrP(23-159) aggregated significantly faster compared with the truncated fragment humPrP(23-144), clearly demonstrating that helix1 is involved in the aggregation process. However, helix1 is not resistant to digestion with proteinase K in fibrillar humPrP(23-159), suggesting that helix1 is not converted to beta-sheet. This is confirmed by Fourier transformation infrared spectroscopy since there is almost no difference in beta-sheet content of humPrP(23-159) fibrils compared with humPrP(23-144). In conclusion, we provide strong direct evidence that in contrast to earlier assumptions helix1 is not converted into beta-sheet during aggregation of PrP(C) to PrP(Sc).

Circular Dichroism↗

PubMed1997

Conformational flexibility and polymerization of vesicular stomatitis virus matrix protein.

The matrix protein of vesicular stomatitis virus (VSV) plays a pivotal role in viral assembly. We previously demonstrated the ability of M protein to self-associate at low salt concentrations. Now, we show the ability of M protein to polymerize in the presence of ZnCl2 in a nucleation-dependent manner. Analysis of kinetics revealed that the nuclei are probably made of three or four molecules of M. These results are consistent with the idea that in vitro self association of M protein is not due to amorphous aggregation but rather reflects an intrinsic ability of M to polymerize. Using attenuated total reflectance Fourier transform infrared spectroscopy, we showed that M polymerization is associated with an increase in the beta-sheet content of the protein. We propose a model explaining both the apparent M protein solubility in infected cells and how M polymerization could promote viral assembly. Data available for other negative strand viruses suggest that M polymerization may be the general basis of viral assembly.

Chlorides↗

PubMed1999

Transdermal delivery of naloxone: ex vivo permeation studies.

The purpose of this investigation was to study the feasibility of transdermal drug delivery of the potent opioid antagonist naloxone. The pharmacokinetic profile of naloxone makes it a suitable candidate for transdermal delivery. Ex vivo permeation of naloxone through excised rat skin was studied using a diffusion cell. Radiochemical assay of drug concentration and the use of rat as an animal model were adopted in this study. Naloxone possesses characteristics favorable to percutaneous absorption: i.e. a low molecular weight (327.37), water solubility and a good lipid-water partition coefficient of 12.94+/-1.29 at pH 7.4. The flux (microg/cm2/h) values varied from 6.59+/-0.72 in control to 27. 18+/-4.26 in dimethyl formamide. The affinity of naloxone to skin in the presence of propylene glycol was decreased by 6.2 times compared to the control. Fourier transform infrared spectroscopy was used to study the effect of various sorption promoters on intercellular lipid pathways in skin. A change in lipid fluidization corresponding to broadening for both C-H symmetric (near 2850 cm-1) and C-H asymmetric (near 2920 cm-1) stretching was observed. An attempt was made to correlate the molecular weight of sorption promoters with skin affinity values of naloxone.

Administration, Cutaneous↗

PubMed2006

Medical application of an infrared free-electron laser: selective removal of cholesterol ester in carotid artery atheromatous plaques.

OBJECT: . The purpose of this study was to determine the effectiveness of infrared free-electron laser (FEL) irradiation of cholesterol esters in human carotid artery (CA) atheromas. METHODS: The degradation of cholesterol ester was estimated from changes in the infrared absorption spectra acquired using microscopic transmission Fourier transform infrared spectroscopy. An FEL emitting radiation at 5.75-microm wavelengths and a power density of 15.9 W/cm2 was used to treat intimal slices of extirpated human arterial atherosclerotic plaques. Peak signals derived from an ester bond of cholesterol ester decreased in height as irradiation time increased and disappeared completely after 180 seconds. No other change was observed in the infrared absorption spectrum after 180 seconds of irradiation, and no histological damage was noted. CONCLUSIONS: The authors concluded that FEL irradiation can remove cholesterol ester selectively from human atheromatous CA plaques. This novel technique differs from previous approaches involving conventional lasers.

Atherosclerosis↗

PubMed2006

Isolation and partial characterization of a biosurfactant produced by Streptococcus thermophilus A.

Isolation and characterization of the surface active components from the crude biosurfactant produced by Streptococcus thermophilus A was studied. A fraction rich in glycolipids was obtained by the fractionation of crude biosurfactant using hydrophobic interaction chromatography. Molecular (by Fourier transform infrared spectroscopy) and elemental compositions (by X-ray photoelectron spectroscopy) were determined. Critical micelle concentration achieved was 20 g/l, allowing for a surface tension value of 36 mJ/m(2). Moreover, this glycolipid rich fraction was found to be an anti-adhesive and antimicrobial agent against several bacterial and yeast strains isolated from explanted voice prostheses. Further purification steps should be carefully analyzed as each purification step will increase the costs and decreases the amounts of biosurfactants recovered.

Bacteria↗

PubMed2006

Preparation of poly epsilon-caprolactone nanoparticles containing magnetite for magnetic drug carrier.

Magnetic poly epsilon-caprolactone (PCL) nanoparticles were prepared in a well shaped spherical form by the o/w emulsion method. The influence of some preparative variables on the size and surface property was investigated. Nanoparticles were smooth, well individualized and homogeneous in size. The presence of magnetite and its superparamagnetic characteristic were confirmed by transmission electron microscope (TEM), Fourier transform infrared spectroscopy (FT-IR) and vibrating sample magnetometer (VSM), respectively. The anti-cancer drug was encapsulated in the magnetic nanoparticle during preparation. A typical release behavior was observed for 30 days. In vitro experiment of magnetic susceptibility under external magnetic field demonstrated that the magnetic PCL nanoparticles have sufficient magnetic susceptibility for a potential magnetic drug carrier for targeted delivery.

Cisplatin↗

PubMed2005

Sorption of the antimicrobial ciprofloxacin to aluminum and iron hydrous oxides.

Solution chemistry (pH, ionic strength (I), and sorbate-to-sorbent ratio) effects on ciprofloxacin sorption to hydrous oxides of Al (HAO) and Fe (HFO) were investigated using macroscopic and spectroscopic analyses. Sorption to both HAO and HFO showed a strong pH-dependent behavior, following the fraction of zwitterionic species over the entire pH range studied. Increase in I from 0.01 to 0.5 M had an insignificant effect on the extent of ciprofloxacin sorption, and isotherms were well-described by the Langmuir model. HFO possessed a higher sorption capacity (0.066 mmol kg(-1)) than HAO (0.041 mmol kg(-1)). Ligand-promoted dissolution of hydrous oxides, more pronounced for HAO, was observed in the presence of ciprofloxacin, but at a fairly high initial concentration (0.5 mM). Attenuated total reflectance Fourier transform infrared spectroscopy analysis indicated that different types of ciprofloxacin surface complexes are formed with HAO and HFO; while a monodentate mononuclear complex (with -COO-) appears likely between ciprofloxacin and HAO, keto O and one O from COO- seem to be involved in the formation of a six-membered ring with Fe on the HFO surface. The study results are expected to increase our understanding of the environmental reactivity of fluoroquinolones, an important class of antimicrobial compounds.

Adsorption↗

PubMed1996

A correlation between thermal stability and structural features of staphylokinase and selected mutants: a Fourier-transform infrared study.

Variants of recombinant staphylokinase (Sak) were investigated by Fourier-transform infrared spectroscopy: Sak (wild type), Sak-M26A, Sak-M26L, and Sak-G34S/R36G/R43H (Sak-B). Estimation of the secondary structure and hydrogen-deuterium exchange experiments revealed the existence of fast-exchanging and strongly solvent-exposed fractions of the helical structures in the two samples Sak and Sak-M26L. These two samples are also thermally less stable with unfolding transition temperatures of 43.7 degrees C (Sak) and 43.5 degrees C (Sak-M26L), respectively. On contrast, Sak-M26A and Sak-G34S/R36G/R43H have a slower hydrogen-deuterium exchange, have a smaller solvent-exposed portion of the helical part, and are more resistant against thermal unfolding; the transition temperatures are 51.7 degrees C and 59.3 degrees C, respectively. The secondary structure analysis was performed by two different approaches, by curve-fitting after band narrowing and by pattern recognition (factor analysis) based upon reference spectra of proteins with known crystal structure. Within the limits of the used methods, we are unable to detect significant differences in the secondary structure of the four variants of Sak. According to the results of the factor analysis, the portions of secondary structure elements were obtained to 16-20% alpha-helix, 28-30% beta-sheet, 23-27% turns, 28-30% irregular (random) and other structure. The sharp differences in the specific plasminogen-activating capacity (Sak, Sak-G34S/R36G/R43H and Sak-M26L are fully active, but Sak-M26A does not form a stable complex with plasminogen) are not reflected in the structural features revealed by the infrared spectra of this study.

Deuterium↗

PubMed2006

Residue formations of phosphorus hydride polymers and phosphorus oxyacids during phosphine gas fumigations of stored products.

With the extent of international usage and the critical role phosphine gas (PH3) plays in commercial pest control, identification of the residual components deposited during fumigation is mandatory. It has been postulated that these infrequent residues are primarily composed of phosphoric acid or reduced forms of phosphoric acid [hypophosphorous acid (H3PO2) and phosphorous acid (H3PO3)], due to the oxidative degradation of phosphine. Using environmental scanning electron microscopy, gas phase Fourier transform infrared spectroscopy, and X-ray fluorescence spectroscopy, the structural elucidation and formation mechanism of the yellow amorphous polyhydric phosphorus polymers (P(x)H(y)) that occur in addition to the lower oxyacids of phosphorus in residues deposited during PH3 fumigations of select tobacco commodities are explored. This research determined that nitric oxide gas (or nitrogen dioxide) initiates residue formation of phosphorus hydride polymers and phosphorus oxyacids during PH3 fumigations of stored products.

Drug Residues↗

PubMed1997

Solid-state investigation of fluocinolone acetonide.

Three crystalline modifications of fluocinolone acetonide, A, B and C, were characterized by means of FTIR, DSC, TG-FTIR, MICRO-FTIR and X-ray diffractometry. They were easily differentiated by their IR absorption bands in the 3600-3400 cm-1 range. The thermal behavior was also elucidated using combined techniques; thermomicroscopy and thermogravimetry coupled with Fourier transform infrared spectroscopy were found to be very useful tools for a better understanding of thermal events. On heating, form A and C were fully converted into form B. Polymorph A was found to be enantiotropically related to B, while form C was monotropically related to B. The existence of three polymorphs was confirmed by means of their different X-ray diffraction patterns. Detailed methods of preparation of the three modifications are also described.

Administration, Topical↗

PubMed1999

Determination of the diglycidyl ether of bisphenol A and its derivatives in canned foods.

Migration of the diglycidyl ether of bisphenol A (DGEBA) to food from can enamels and can pull-top seals is reported. Derivatives of DGEBA are also determined in some foods. Levels of DGEBA in the foods surveyed in this study range from nondetected (<0.3 ppb) to 50 mg/kg as determined by liquid-liquid extraction or solid-phase extraction coupled with high-pressure liquid chromatography using fluorescence detection. Confirmation of the analytes is by gas and/or liquid chromatography with mass spectral analysis. Fourier transform infrared spectroscopy with 30 degrees specular reflectance/transmittance is used to characterize the coated food contact surfaces. Stability studies with DGEBA in water, acid, and saline solutions show conversion to the hydrolysis products and chloro adducts occurs readily. The presence of DGEBA derivatives in food demonstrates that analysis for DGEBA migration alone is not a good indicator of total migration from can coatings to foods.

Animals↗

PubMed2000

Role of the carboxy terminus of Escherichia coli FtsA in self-interaction and cell division.

The role of the carboxy terminus of the Escherichia coli cell division protein FtsA in bacterial division has been studied by making a series of short sequential deletions spanning from residue 394 to 420. Deletions as short as 5 residues destroy the biological function of the protein. Residue W415 is essential for the localization of the protein into septal rings. Overexpression of the ftsA alleles harboring these deletions caused a coiled cell phenotype previously described for another carboxy-terminal mutation (Gayda et al., J. Bacteriol. 174:5362-5370, 1992), suggesting that an interaction of FtsA with itself might play a role in its function. The existence of such an interaction was demonstrated using the yeast two-hybrid system and a protein overlay assay. Even these short deletions are sufficient for impairing the interaction of the truncated FtsA forms with the wild-type protein in the yeast two-hybrid system. The existence of additional interactions between FtsA molecules, involving other domains, can be postulated from the interaction properties shown by the FtsA deletion mutant forms, because although unable to interact with the wild-type and with FtsADelta1, they can interact with themselves and cross-interact with each other. The secondary structures of an extensive deletion, FtsADelta27, and the wild-type protein are indistinguishable when analyzed by Fourier transform infrared spectroscopy, and moreover, FtsADelta27 retains the ability to bind ATP. These results indicate that deletion of the carboxy-terminal 27 residues does not alter substantially the structure of the protein and suggest that the loss of biological function of the carboxy-terminal deletion mutants might be related to the modification of their interacting properties.

Alleles↗

PubMed1998

[FTIR study on structure of pearl and nacre of mollusk shell].

Pearl and nacre of mollusk shell (Hyriopsis cumingii) were studied with high resolution Fourier transformation infrared spectroscopy (FTIR) and X-ray diffraction (XRD). The results of X-ray diffraction indicate that the samples are still aragonite after thermal treatment at 160 degrees C for 5hr, however, the significant variations of peak position and band width of FTIR spectra reveal the interaction between organic matrix and inorganic crystal in biomineralized materials. The variations of spectra were studied by a curve-fitting algorithm and the band related to the interaction is revealed.

Animals↗

PubMed1994

Structural studies of tau protein and Alzheimer paired helical filaments show no evidence for beta-structure.

We have studied the conformation of tau protein and Alzheimer paired helical filaments (PHF) by several spectroscopic, scattering, and imaging methods revealing the overall shape and the conformation of the polypeptide backbone. Tau protein behaves in solution as if it were denatured; no evidence for compact folding was detected. The protein is highly extended, there is no defined radius of gyration, and the scattering is best described by that of a random ("Gaussian") polymer. CD and Fourier transform infrared spectroscopy show only a minimal content of ordered secondary structure (alpha-helix or beta-sheet). Similarly, PHFs from Alzheimer brain tissue show no detectable secondary structure by x-ray diffraction or spectroscopy. It is thus unlikely that the aggregation of tau into Alzheimer PHFs is based on interactions between strands of beta-sheets (a model currently favored for other disease-related polymers such as beta-amyloid fibers of Alzheimer's disease).

Alzheimer Disease↗

PubMed2005

Alginate microparticles prepared by spray-coagulation method: preparation, drug loading and release characterization.

A spray-coagulation method was developed for the preparation of large scale of porous alginate microparticles. The effect of three variables on porosity was evaluated: (1) alginate solution concentration (2) the concentration of CaCl2 in the coagulation medium and (3) the ratio of guluronic acid to manuronic acid of the alginate. Methylene blue (MB), a highly water-soluble compound and a practically water-insoluble compound, 4-phenylazoaniline (PAA) were used as the model drugs to study drug loading and release characteristics from alginate microparticles. The release of the model compounds from the microparticles was found to depend upon the release medium. Incomplete in vitro release of both model drugs in deionized (DI) water was observed. The release of MB in simulated gastrointestinal fluid (0.1N HCl) was fast and complete, while the release of PAA was slow in 0.1N HCl and fast in phosphate buffer solution (pH 6.8). Interactions between the model drugs and alginate microparticles were identified from scanning electron microscopy (SEM), differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FT-IR) analysis. The results indicated that (1) porous alginate microparticles can be produced by the spray-coagulation method; (2) drugs can be loaded by the adsorption method; (3) and the obtained microparticles may be used for delaying the release of drugs of low water solubility in acidic conditions.

Adsorption↗

PubMed2006

Self-sustained kinetic oscillations in CO oxidation over silica-supported Pt.

Isothermal self-sustained kinetic oscillations in CO oxidation over silica-supported Pt at near-atmospheric pressure were studied by combined in situ Fourier transform infrared spectroscopy and mass spectrometry. The use of a specially designed reactor and careful choice of the physical properties of the catalyst and reaction conditions made it possible to eliminate diffusion limitations, to determine the maximum CO oxidation rate per Pt site in the purely kinetic regime and to clarify the mechanism of the oscillations. Specifically, our results indicate that during the high reactive periods the reaction mainly occurs on the oxide surface.

Carbon Monoxide↗

PubMed2003

A novel lytic peptide composed of DL-amino acids selectively kills cancer cells in culture and in mice.

The high toxicity of most chemotherapeutic drugs and their inactivation by multidrug resistance phenotypes motivated extensive search for drugs with new modes of action. We designed a short cationic diastereomeric peptide composed of d- and l-leucines, lysines, and arginines that has selective toxicity toward cancer cells and significantly inhibits lung metastasis formation in mice (86%) with no detectable side effects. Its ability to depolarize the transmembrane potential of cancer cells at the same rate (within minutes) and concentration (3 micro m), at which it shows biological activity, suggests a killing mechanism that involves plasma membrane perturbation. Confocal microscopy experiments verified that the cells died as a result of acute injury, swelling, and bursting, suggesting necrosis. Biosensor binding experiments and attenuated total reflectance-Fourier transform infrared spectroscopy using model membranes have substantiated its high selectivity toward cancer cells. Although this is an initial study that looked at tumor formation rather than the ability of the peptides to reduce established tumors, the simple sequence of the peptide, its high solubility, substantial resistance to degradation, and inactivation by serum components might make it a good candidate for future anticancer treatment.

3T3 Cells↗

PubMed1993

Fluoridated carbonatoapatite in the intermediate layer between glass ionomer and dentin.

The intermediate layer between a glass ionomer restoration and dentin was examined morphologically by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). SEM showed strong bonding between the dentin and glass ionomer and formation of an intermediate layer between them. FTIR spectra of successive scrapings through the ionomer/dentin interface zone showed that the intermediate layer is composed primarily of mineral fluoridated carbonatoapatite. The presence of this sparingly soluble mineral at the interface between the tooth and the restoration may provide high resistance to secondary caries and may, thus, be of great clinical importance.

Apatites↗