Amyloid deposition and CBF patterns predict conversion of mild cognitive impairment to dementia.

Mild cognitive impairment (MCI) can include the transition from a normal state to dementia. To explore biomarkers for the development of dementia, we performed an 18-month follow-up study in 28 patients with amnestic MCI. Amyloid deposition was examined using PiB PET, and cerebral blood flow (CBF) was examined using SPECT. Cognitive function was periodically assessed. The rate of conversion to dementia was higher in the PiB-positive/equivocal group (74%) than in the PiB-negative group (33%) (p = 0.041). Perfusion SPECT was performed in 16 patients. MCI patients with an AD-characteristic pattern of reduced CBF had a higher PiB-positive/equivocal rate (82%) than those with a non-AD pattern (20%) (p = 0.018), and patients with an AD pattern had a higher conversion rate (82%) than those with a non-AD pattern (40%) (p = 0.094). Clinically, all PiB-positive converters were diagnosed as having Alzheimer's disease (AD), whereas PiB-negative converters were thought to have some form of dementia other than AD. Amyloid PET is useful for predicting conversion to AD in MCI patients. A pattern analysis of perfusion SPECT findings might also be helpful for predicting conversion to AD, but with a lower specificity.

Nanoarchitectonics for Improving Catalytic Performance of α-Alumina-supported Gold Nanoflower by Water Extraction and Ultraviolet-Ozone Treatment.

Shape-controlled nanocrystals, such as nanoflowers, are expected to serve as innovative nanocatalysts with high catalytic activity. It is well-established that these nanocrystals can be readily synthesized with specific shapes using colloidal methods in solutions containing capping agents. However, these capping agents tend to reduce the catalytic activity of nanocatalysts. Therefore, it is imperative to remove these agents without altering the morphology to enhance catalytic efficiency. In this study, we developed a method for eliminating melamine, a common capping agent, from supported Au nanoflowers using water extraction and ultraviolet-ozone treatment. This process significantly enhances the catalytic performance, particularly for alcohol oxidation reactions such as the conversion of 1-phenylethyl alcohol to acetophenone.

Alterations in the detergent-induced membrane permeability and solubilization of saturated phosphatidylcholine/cholesterol liposomes: effects of poly(ethylene glycol)-conjugated lipid.

We have investigated the effects of two bile salts, chenodeoxycholate (CDC) and ursodeoxycholate (UDC), and a widely used detergent, Triton X-100 (T(X-100)), on normal and poly(ethylene glycol)-modified liposomes (PEGylated liposomes). We tested various lipid compositions, including hydrogenated soybean phosphatidylcholine/cholesterol/PEG-conjugated lipid (HSPC/PEG-lipid). Alterations in permeability were determined by the rate of drug release from the liposomes and solubilization was assessed by measuring the particle size of liposomes. In addition, we attempted to observe interactions between the detergents and lipid bilayers by using surface plasmon resonance (SPR). CDC induced drug release from liposomes in a dose-dependent manner, and the PEGylated liposomes tended to be susceptible to CDC. While UDC did not strongly induce drug release from liposomes, UDC exhibited a similar tendency with CDC. In case of T(X-100), there were significant differences in the percentage of released drug between normal and PEGylated liposomes, and the percentage of T(X-100)-induced drug release further increased with an increased ratio of PEG-lipid. SPR analysis revealed that the lipid bilayer including PEG-lipid was selectively solubilized by T(X-100), correlating with the drug release data. These results suggest that the effect of detergents on the lipid bilayer of liposomes depends on both the kind of detergent and the lipid composition, including the presence or absence of PEG-lipid. Moreover, the effects of T(X-100) on the lipid bilayers of the PEGylated liposomes significantly differed from those on the lipid bilayers of the normal liposomes.

Comparison of particle size and dispersion state among commercial cyclosporine formulations and their effects on pharmacokinetics in rats.

Generic versions of Neoral, a microemulsion capsule formulation of cyclosporine, have been approved worldwide. However, there are concerns about the quality and efficacy of the generics due to the formulation specificity and differences in inactive ingredients among products. In this study, we measured the physicochemical properties of both the innovator and the generic formulations, and compared their bioavailability in rats. When the capsule contents were dispersed in water, the absorbance (600 nm wavelength) of generic products was higher than that of the innovator. Whereas the dispersion solution of the innovator in Fed State Simulated Intestinal Fluid was nearly clear, that of all the generics became white and turbid. The mean diameter of the microemulsion (or emulsion) formed in water by the generics was 39.7, 57.7, 64.5, and 74.8 nm, all of which were larger than that of the innovator (26.4 nm). Although the T(max) of the generics tended to be long relative to that of the innovator, there were no significant differences between the innovator and generics with regard to maximum blood concentration (C(max)) or area under the curve (AUC). These results suggest that the physicochemical differences between the innovator and the generics will not have a significant effect on C(max) or AUC, which is necessary to ensure bioequivalence.

Preparing Alumina-Supported Gold Nanowires for Alcohol Oxidation.

The development of shape-controlled noble metal nanocrystals such as nanowires (NWs) is progressing steadily owing to their potentially novel catalytic properties and the ease with which they can be prepared by reducing the metal ions in a particular solution as capping agents. Recently, many reports have been presented on the preparation of shape-controlled Au nanocrystals, such as nanostars and nanoflowers, by a one-pot method using 2-[4-(2-hydroxyethyl)-1-piperazinyl] ethanesulfonic acid (HEPES) as capping and reducing agents. The catalytic activity is depressed due to the adsorption of the capping agent onto a Au surface. Since HEPES has low binding affinities on the Au surface, shape-controlled nanocrystals obtained using HEPES are effective for application as nanocatalysts because HEPES was easily removed from the Au surface. In this study, we report the preparation of AuNWs, with an average diameter of 7.7 nm and lengths of a few hundred nanometers, in an aqueous solution containing HEPES and sodium borohydride. A γ-Al2O3-supported AuNW (AuNW/γ-Al2O3) catalyst was obtained using catalytic supporters and a water extraction method that removed HEPES from the Au surface without morphological changes. AuNW/γ-Al2O3 was then utilized to catalyze the oxidation of 1-phenylethyl alcohol to acetophenone. The formation rate of acetophenone over AuNW/γ-Al2O3 was 3.2 times that over γ-Al2O3-supported spherical Au nanoparticles (AuNP/γ-Al2O3) with almost the same diameter.

Magnetic Fe3O4-Supported Gold Nanoflowers with Lattice-Selected Surfaces: Preparation and Catalytic Performance.

Nanoflowers (NFs)-shape-controlled noble metal nanocrystals-have garnered significant attention because of their novel catalytic properties and applicability. In this paper, we report the preparation and catalytic performance of a magnetic Fe3O4-supported AuNF catalyst with a clean surface. The magnetically supported AuNFs were obtained by using magnetic Fe3O4 as the support. However, when nonmagnetic γ-Al2O3 was utilized as the support, the AuNFs did not exhibit a magnetic response. These supported AuNFs were utilized to catalyze the oxidation of 1-phenylethyl alcohol to acetophenone using air (1 atm) as the oxidant. The rate of formation of acetophenone using supported AuNFs was 8-fold higher than that of acetophenone using supported spherical Au nanoparticles of comparable size. In addition, the Fe3O4-supported AuNFs exhibited a higher rate of formation of acetophenone than the Al2O3-supported AuNFs. The Fe3O4-supported AuNFs were recovered using a magnet, and the recovered catalyst was reused under identical catalytic reaction conditions. The rate of formation of acetophenone using recovered Fe3O4-supported AuNFs remained unchanged, demonstrating no loss of catalytic activity.

Lectin affinity capture, isotope-coded tagging and mass spectrometry to identify N-linked glycoproteins.

We describe here a strategy for the large-scale identification of N-glycosylated proteins from a complex biological sample. The approach, termed isotope-coded glycosylation-site-specific tagging (IGOT), is based on the lectin column-mediated affinity capture of a set of glycopeptides generated by tryptic digestion of protein mixtures, followed by peptide-N-glycosidase-mediated incorporation of a stable isotope tag, 18O, specifically into the N-glycosylation site. The 18O-tagged peptides are then identified by multi-dimensional liquid chromatography-mass spectrometry (LC-MS)-based technology. The application of this method to the characterization of N-linked high-mannose and/or hybrid-type glycoproteins from an extract of Caenorhabditis elegans proteins allowed the identification of 250 glycoproteins, including 83 putative transmembrane proteins, with the simultaneous determination of 400 unique N-glycosylation sites. Because the method is applicable to the systematic identification of a wide range of glycoproteins, it should facilitate basic glycobiology research and may be useful for diagnostic applications, such as genome-wide screening for disease-related glycoproteins.

Cognitive impairment and neurovascular function in patients with severe steno-occlusive disease of a main cerebral artery.

OBJECTIVE: Patients with severe steno-occlusive disease of a main cerebral artery may demonstrate cognitive impairment without identification of causative lesions on magnetic resonance imaging. We investigated whether cognitive impairment in these patients is associated with regional cerebral blood flow (rCBF), leukoaraiosis, risk factors of atherosclerosis and cerebrovascular reserve (CVR), which shows so-called clinical neurovascular function. METHODS: In 65 patients with severe steno-occlusive disease of an internal carotid artery or a middle cerebral artery (MCA) and no cerebral infarction (CI), we examined cognitive function with COGNISTAT, grades of leukoaraiosis, and CBF and CVR as calculated by iodine-123-N-isopropyl-p-iodoamphetamine single photon emission computed tomography and blood data. We compared such values of the left and right sided diseases. rCBF and CVR on the affected side were compared to other side. RESULTS: Logistic regression analysis revealed that CVR correlated with cognitive impairment. There was no significant difference in rCBF, CVR, or COGNISTAT score when comparing the left and right sided diseases. There were good correlations between CBF or CVR of the ipsilateral MCA area and ipsilateral and contralateral other areas. CONCLUSION: Cognitive impairment is associated with CVR in the whole brain. Nonselective widespread neurovascular mild dysfunction can be a reason for cognitive impairment in patients with severe steno-occlusive disease of a main cerebral artery and no CI.

Pharmaceutical quality evaluation of lipid emulsions containing PGE1: alteration in the number of large particles in infusion solutions.

There are two generics of a parenteral lipid emulsion of prostaglandin E1 (PGE(1)) (Lipo-PGE(1)) in addition to two innovators. It was reported the change from innovator to generic in clinical practice caused the slowing of drip rate and formation of aggregates in the infusion line. Thus, we investigated the difference of pharmaceutical quality in these Lipo-PGE(1) formulations. After mixing with some infusion solutions, the mean diameter and number of large particles were determined. Although the mean diameter did not change in any infusion solutions, the number of large particles (diameter >1.0 microm) dramatically increased in generics with Hartmann's solution pH 8 or Lactec injection with 7% sodium bicarbonate. Next, we investigated the effect of these infusion solutions on the retention rate of PGE(1) in lipid particles. The retention rate of PGE(1) in these two infusion solutions decreased more quickly than that in normal saline. Nevertheless, there were no significant differences among the formulations tested. Our results suggest that there is no difference between innovators and generics except in mixing with these infusion solutions. Furthermore, that monitoring the number of large particles can be an effective means of evaluating pharmaceutical interactions and/or the stability of lipid emulsions.

Multiplexed two-dimensional liquid chromatography for MALDI and nanoelectrospray ionization mass spectrometry in proteomics.

We developed a multiplexed two-dimensional separation system based on reversed phase (RP)--strong cation exchange (SCX) chromatography as a front-end device for matrix-assisted laser desorption ionization (MALDI) or nanoelectrospray ionization (nanoESI) mass spectrometry. Tryptic peptide mixtures were fractionated on a reversed-phase HPLC column, and each fraction was loaded onto multiplexed SCX microcolumns. Because this second chromatography was carried out in parallel, the analysis time is independent of the fraction number in the first RP-HPLC separation. The resultant samples were desalted/concentrated and eluted onto a MALDI plate with matrix-containing elution solutions in parallel, or eluted with optimized solutions for nanoESI and loaded onto nanoESI sprayers by an automated instrument. The soluble portion of HCT116 lysate was digested and fractionated using a 48-plexed chromatography system. Approximately 1000 unique peaks were detected in MALDI-MS with 3000 MS/MS spectra, while 724 peptides with ultrahigh peptide mass accuracy (sub-ppm error) were identified in nanoESI-FTICR mass spectrometry with five integrated selected ion monitoring scans. Since MS measurement with this off-line LC-LC approach is not restricted by continuous LC elution, it is expected to be useful especially in cases where repeated analysis with different scan modes or long-term data acquisition is required.

Large-scale identification of Caenorhabditis elegans proteins by multidimensional liquid chromatography-tandem mass spectrometry.

A proteome of a model organism, Caenorhabditis elegans, was analyzed by an integrated liquid chromatography (LC)-based protein identification system, which was constructed by microscale two-dimensional liquid chromatography (2DLC) coupled with electrospray ionization (ESI) tandem mass spectrometry (MS/MS) on a high-resolution hybrid mass spectrometer with an automated data analysis system. Soluble and insoluble protein fractions were prepared from a mixed growth phase culture of the worm C. elegans, digested with trypsin, and fractionated separately on the 2DLC system. The separated peptides were directly analyzed by on-line ESI-MS/MS in a data-dependent mode, and the resultant spectral data were automatically processed to search a genome sequence database, wormpep 66, for protein identification. The total number of proteins of the composite proteome identified in this method was 1,616, including 110 secreted/targeted proteins and 242 transmembrane proteins. The codon adaptation indices of the identified proteins suggested that the system could identify proteins of relatively low abundance, which are difficult to identify by conventional 2D-gel electrophoresis (GE) followed by an offline mass spectrometric analysis such as peptide mass fingerprinting. Among the approximately 5,400 peptides assigned in this study, many peptides with post-translational modifications, such as N-terminal acetylation and phosphorylation, were detected. This expression profile of C. elegans, containing 571 hypothetical gene products, will serve as the basic data of a major proteome set expressed in the worm.