Synthesis and biological evaluation of [18F]fluorovinpocetine, a potential PET radioligand for TSPO imaging.

Despite of various PET radioligands targeting the translocator protein TSPO 18-KDa are used for the investigations of neuroinflammatory conditions associated with neurological disorders, development of new TSPO radiotracers is still an active area of the researches with a major focus on the 18F-labelled radiotracers. Here, we report the radiochemical synthesis of [18F]vinpocetine, fluorinated analogue of previously reported TSPO radioligand, [11C]vinpocetine. Radiolabeling was achieved by [18F]fluoroethylation of apovincaminic acid with [18F]fluoroethyl bromide. [18F]vinpocetine was obtained in quantities >2.7 GBq in RCY of 13% (non-decay corrected), and molar activity >60 GBq/µmol within 95 min synthesis time. Preliminary PET studies in a cynomolgus monkey and metabolite studies by HPLC demonstrated similar results by [18F]vinpocetine as for [11C]vinpocetine, including high blood-brain barrier permeability, regional uptake pattern and fast washout from the NHP brain. These results demonstrate that [18F]fluorovinpocetine warrants further evaluation as an easier accessible alternative to [11C]vinpocetine.

High molecular weight organic compounds (HMW-OCs) in severe winter haze: Direct observation and insights on the formation mechanism.

High molecular weight organic compounds (HMW-OCs), formed as secondary organic aerosols (SOA), have been reported in many laboratory studies. However, little evidence of HMW-OCs formation, in particular during winter season in the real atmosphere, has been reported. In January 2013, Beijing faced historically severe haze pollution, in which the hourly PM2.5 concentration reached as high as 974 µg m-3. Four typical haze events (HE1 to HE4) were identified, and HE2 (Jan. 9-16) was the most serious of these. Based on the hourly observed chemical composition of PM2.5 and the daily organic composition analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), we found that abundant ion peaks in m/z 200-850 appeared on heavy haze days, whereas these were negligible on a clear day, indicating the existence of HMW-OCs in the wintertime haze. A negative nonlinear correlation between HMW-OCs and O3 suggested that gas oxidation was not likely to be the dominant mechanism for HMW-OCs formation. During the heavy haze events, the relative humidity and mass ratio of H2O/PM2.5 reached as high as 80% and 0.2, respectively. The high water content and its good positive correlation with HMW-OCs indicated that an aqueous-phase process may be a significant pathway in wintertime. The evidence that acidity was much higher during HE2 (0.37 µg m-3) than on other days, as well as its strong correlation with HMW-OCs, indicated that acid-catalyzed reactions likely resulted in HMW-OCs formation during the heavy winter haze in Beijing.

Progesterone inhibits apolipoprotein-mediated cellular lipid release: a putative mechanism for the decrease of high-density lipoprotein.

In order to investigate the mechanism for female gonadal hormones to regulate the plasma high-density lipoprotein (HDL) level, the effect of 17 beta-estradiol and progestogens was examined in vitro on the assembly of HDL by free apolipoprotein A-I (apoA-I) with cellular cholesterol and phospholipid. ApoA-I generated HDL particles by removing cholesterol and phospholipid from human fibroblasts, MRC-5. While 17 beta-estradiol did not influence this reaction, progesterone suppressed the removal by apoA-I of both cholesterol and phospholipid, with the extent of the inhibition more for cholesterol than phospholipid. Three other synthetic progestogens showed the similar inhibitory effect on the cellular cholesterol release. Cellular cholesterol de novo-synthesized from mevalonolactone entered more into the acyl-esterified cholesterol compartment and less to the unesterified compartment in the presence of progesterone. On the other hand, progesterone did not influence the overall mass ratio of free and esterified cholesterol in the cell. Cell-surface cholesterol was also uninfluenced by progesterone when probed by extracellular cholesterol oxidase reaction or by diffusion-mediated cellular cholesterol release to cyclodextrin. Neither caveolin-1 nor ABCA1 expression was influenced by progesterone. Progesterone thus seems primarily to alter the specific intracellular cholesterol compartment that is related to the apoA-I-mediated HDL assembly. This mechanism might contribute to the decrease of plasma HDL by administration of progestogen in women under hormone replacement therapy.

High-energy collision-induced dissociation of small polycyclic aromatic hydrocarbons.

High-energy collision-induced dissociation (CID) experiments on polycyclic aromatic hydrocarbons (PAHs) having 2-6 rings, naphthalene, anthracene, phenanthrene, fluoranthene, pyrene and coronene, were performed, and the relative abundances of their fragment ions were investigated as a function of collision energy. The results revealed that the PAHs except naphthalene showed a bimodal-type distribution of positive fragmentation ions, which is closely similar to the fragment-ion distribution reported for the CID of three-dimensional fullerene, C(60)(+) and C(70)(+). The three-ring isomers of anthracene and phenanthrene and the four-ring isomers of fluoranthene and pyrene can be distinguishable in their spectra under an electron ionization energy of 70 eV, but the high-energy CID spectra of the three- and four-ring isomers were almost identical. The fragmentation corresponding to fragment ions in the low-mass region of the bimodal CID spectra could be interpreted by the simple statistical model that fragment ions are formed by random evaporation from the molecular ions after a considerable structural rearrangement, 'phase transition', occurring at some high-energy state.

Microembolic signals and diffusion-weighted MR imaging abnormalities in acute ischemic stroke.

BACKGROUND AND PURPOSE: The clinical significance of microembolic signals (MESs) detected by transcranial Doppler sonography (TCD) in acute ischemic stroke remains unclear. The purpose of the present study was to assess the findings of diffusion-weighted MR imaging (DWI) and other clinical characteristics in patients with acute ischemic stroke and MESs. METHODS: We performed TCD and DWI within 48 hours and 7 days, respectively, after stroke onset in 28 patients with acute brain infarction. The relationship between the number of MESs and DWI findings, risk factors for stroke, National Institutes of Health Stroke Scale (NIHSS) score on admission, and arterial disease was examined. RESULTS: Ten patients had MESs detected by TCD (MES group) and 18 had no MESs (control group). The frequency of hypertension, diabetes mellitus, hyperlipidemia, and smoking; NIHSS score; blood-coagulation parameters; and interval between stroke onset and DWI study did not differ between the two groups. However, arterial disease was more frequent in the MES group than in the control group. Small, multifocal ischemic lesions (<10 mm in diameter) on DWI were more frequent in the MES group than in the control group. Conventional CT and MR imaging often failed to show these lesions. CONCLUSION: Small, often asymptomatic DWI abnormalities were more frequent in patients with MESs detected by TCD and with large-vessel occlusive diseases than in stroke patients without MESs. TCD and DWI may provide early clues to the mechanism of stroke in the acute phase.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of self-assembled monolayers of ruthenium complexes on gold

Self-assembled monolayers (SAMs) of three ruthenium complexes, [Ru(L)(2)](PF(6))(2), [Ru(L)(tpyPO(3))](PF(6))(2), and [Ru(L18)(tpyPO(3))](PF(6))(2), were prepared on evaporated gold films on glass or stainless steel plates; where L = 2, 6-bis(benzimidazoyl)pyridine, tpyPO(3) = 2,6-bis(2,2':6', 2"-terpyridyl)pyridine phosphanate, and L18 = 2, 6-bis(N-octadecylbenzimidazoyl)pyridine. Structures of these SAM complexes were studied by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). The SAMs were either prepared by direct binding of Ru-complexes to Au films by alkanethiol or by the multilayer method. In the multilayer method 1,4-thiobutylphosphate was used to form a base layer on an Au film, and the base layer was then chemically bridged to the Ru-complexes by zirconium phosphate. MALDI-TOFMS of SAM1, that had been prepared by direct binding of [Ru(L)(2)](PF(6))(2) to the Au film by an octanethiol group, showed cleavage at the S-Au linkages and elimination of the counter anion to yield a molecular ion and its dimeric ion. On the other hand, SAM2 and SAM3, which had been prepared by bridging Ru-complexes [Ru(L)(tpyPO(3))](PF(6))(2) or [Ru(L18)(tpyPO(3))](PF(6))(2) to the base layers with zirconium phosphate, showed dissociation from the base layers and elimination of the counter anion to give ions of the Ru complex molecules and their fragmentation ions. No molecular ion containing the base layer resulting from the S-Au bond cleavage was observed. Copyright 2000 John Wiley & Sons, Ltd.

Cyclical transmission of Plasmodium berghei (Coccidiida: plasmodiidae) by Anopheles omorii (Diptera: Culicidae).

Anopheles omorii, a tree-hole breeding anopheline collected in Japan, transmitted a rodent malaria, Plasmodium berghei ANKA strain, in the laboratory. Mice with 0.2-15% parasitemia, 0.01-1.5% gametocytemia, and 0.001-0.5% exflagellations were used as infective hosts. Oocyst numbers ranged from 2 to 171 (mean 44) on the midgut 8-16 d after the blood meal. Several to hundreds of sporozoites were detected in the salivary glands 14 d after feeding. The mosquitoes were infective to mice from 13 to 40 d after feeding. An. omorii occurs naturally at temperatures less than 25 degrees C and therefore is a suitable laboratory vector for the cyclic transmission of P. berghei, a malaria parasite that completes sporogony at 18-21 degrees C.

[A case of welder presenting with parkinsonism after chronic manganese exposure].

A 56-year-old welder working for 30 years developed postural instability and writing clumsiness since October, 1998. Neurologic findings revealed dystonia of the bilateral shoulders and distal four limbs as well as parkinsonism such as masked face, bradykinesia, rigidity, and retropulsion. Brain MRI showed hyperintensity lesions on T1-weighted images in the bilateral globus pallidus, midbrain, pontine tegmentum, dentate nucleus and cerebral white matter, which reduced in size and density after ten months. The diagnosis of manganese poisoning was made by the high manganese levels of both serum and urine, and by the marked elevated urinary manganese level after administration of the cheleting agent. We pointed out the diagnostic significance of brain MRI in patients with chronic manganese exposure.

[Immunochemical quantification of the airborne chironomid allergens].

We collected airborne particles in the air using a high-volume air sampler, and measured chironomid allergen in the air-filter extract by use of RAST inhibition assay. The results obtained were as follows: 1) The assay revealed the presence of chironomid allergen in the air varying from 6.8 to less than 0.3 ng/m3. 2) In Toyama, seasonal fluctuations of airborne chironomid allergen demonstrated some large peaks in summer (June to August), and showed a tendency to decrease in winter. 3) There were significant correlations between airborne chironomid allergen and dry weight of chironomid midges, collected by using light traps, in all period of a week just before, 1 to 2 weeks before, 2 to 3 weeks before, or 3 to 4 weeks before air sampling. There were not significant correlations between airborne chironomid allergen and weathers (temperature, wind speed and precipitation) during or just before air sampling. 4) In Toyama, the frequency of asthma attacks in the asthmatic children with positive C.p. RAST was much increased in summer compared with that of the negative, in which the amount of airborne chironomid allergen was increased. These results show that there is large amount of chironomid allergen in the air from spring to autumn, especially large in summer, and it is related for some asthmatics to the increase of the frequency of asthma attack.

Oxidation of Benzyl Alcohol with Cu(II) and Zn(II) Complexes of the Phenoxyl Radical as a Model of the Reaction of Galactose Oxidase.

Profound insights into the catalytic mechanism of galactose oxidase (GO) are offered by new models of the active form of the metalloenzyme. The important role of the Cu(II) center in the oxidation of benzyl alcohol to benzaldehyde by the Cu(II)-phenoxyl radical complex of ligand 1 has been revealed by comparison with the reactivity of the corresponding Zn(II)-phenoxyl radical complex; py=2-pyridyl.

N,N-Dimethylformamide-stabilized gold nanoclusters as a catalyst for the reduction of 4-nitrophenol.

In this study, we investigated the catalytic properties of N,N-dimethylformamide (DMF)-stabilized gold nanoclusters (AuNCs) in the reduction of 4-nitrophenol (PNP) to 4-aminophenol by NaBH(4), a well known model reaction to be catalyzed by metal surfaces. The DMF-stabilized AuNCs were prepared in DMF by a surfactant-free method. The DMF-stabilized AuNCs showed high catalytic activity even when used in small quantities (∼10(-7) g). The pseudo-first-order rate constant (k(app)) and activation energy were estimated to be 3 × 10(-3) s(-1) and 31 kJ mol(-1), respectively, with 1.0 µM of the gold catalyst at 298 K. The catalytic activity of the DMF-stabilized AuNCs was strongly influenced by the layer of adsorbed DMF on the Au NCs. This layer of adsorbed DMF prohibited the reactants from penetrating to the surface of the AuNCs via the diffusion at the beginning of the reaction, resulting in an induction time (t(0)) before PNP reduction began. Restructuring of the DMF layer (essentially a form of activation) was the key to achieving high catalytic activity. In addition, atomically monodisperse Au(25)(SG)(18)NCs (SG: glutathione) showed higher catalytic activity in the PNP reduction (k(app) = 8 × 10(-3) s(-1)) even with a low catalyst concentration (1.0 µM), and there was no induction time (t(0)) in spite of the strongly binding ligand glutathione. This suggested that the catalytically active surface sites of the Au(25)(SG)(18)NCs were not sterically hindered, possibly because of the unique core-shell-like structure of the NCs. Retaining these open sites on AuNCs may be the key to making the NCs effective catalysts.

Effects of spiracle-blocking insecticides and microbial insecticides on the predator mirid bug, Nesidiocoris tenuis (reuter) (heteroptera: miridae).

Spiracle-blocking insecticides and microbial insecticides are widely used for Integrated Pest Management (IPM) in Japan while Nesidiocoris tenuis is used for the control of thrips and whiteflies in Kochi Prefecture, Japan. However, the effects of the insecticides mentioned above on N. tenuis were unclear. This study investigated the effects of five spiracle-blocking insecticides and two microbial insecticides on the nymphs and adults ofN. tenuis. Propylene glycol fatty acid monoester was slightly harmful to both the nymphs and adults. Hydroxypropyl starch was slightly harmful to the nymphs, while sodium oleate was slightly harmful to the adults. Decanoyloctanoylglycerol and hydrogenated starch hydrolysate were not harmful to either the nymphs or adults. Beauveria bassiana was extremely harmful to the adults and was moderately harmful to the nymphs. Lecanicillium muscarium was slightly harmful to the adults. Therefore, decanoyloctanoylglycerol and hydrogenated starch hydrolysate can be used in combination with N. tenuis to establish an IPM program.

Development of Plasmodium berghei ookinetes to young oocysts in vitro.

The mosquito stage of Plasmodium berghei was cultivated in vitro, with special attention to ookinete transformation into early oocyst. The ookinetes were obtained by in vitro culture of gametocytes taken from infected mice, purified by density gradient of metrizoic acid or a lymphocyte separation medium, and incubated either in acellular culture or in co-cultivations with mosquito cells. In acellular culture, the ookinetes were found to aggregate with each other and transformed from banana to round shapes. Their inner pellicular membranes and subpellicular microtubules partially disappeared, indicating that development to early oocyst had occurred. Co-cultivation wtih Aedes albopictus cells (C6/36 clone) revealed that ookinetes transformed into early oocyst in the medium, or invaded the cells and then transformed to early oocysts within the cell cytoplasm as well. However all of these transformed cells failed to develop further, i.e., neither deposition of the oocyst capsule nor nuclear division was observed. Many ookinetes which failed to penetrate the Aedes cells were phagocytized within three days of culture. A significant difference between invaded and transformed oocysts and phagocytized ookinetes was seen in that the former lacked vacuole membrane. Co-cultivation with Toxorhynchites amboinensis cells (TRA-284-SFG clone) permitted transformation of ookinetes into early oocysts in the medium as in the acellular culture, but no ookinete invasion nor phagocytosis by the cell was observed.

Penetration of the mosquito midgut wall by the ookinetes of Plasmodium yoelii nigeriensis.

The penetration route of ookinetes of Plasmodium yoelii nigeriensis in the midgut of a mosquito, Anopheles omorii, was investigated by electron microscopy. Within 15-18 h after an infective blood meal, ookinetes could be seen in the midgut lumen, in the process of entering the midgut wall, or lodged between the basement membrane and the basal lamina. The morphology of the ookinetes and their transformation into early oocysts were found to be similar to those previously reported. Ookinetes penetrated the midgut wall by the intercellular route; however, the intracellular occurrence of the parasite was also observed. Vacuoles appeared around the penetrating ookinetes during the penetration process, but no change in the electron density of the rhoptry-microneme complex was noted.

Effects of neurotransmitters or drugs on the in vivo release of dopamine and its metabolites.

The effects of neurotransmitters or drugs on the release of endogenous dopamine (DA) and extracellular levels of its metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), were examined in vivo by intracerebral dialysis. A dialysis tube was implanted stereotaxically through bilateral caudate nuclei of rats and perfused with the Ringer solution. Amounts of DA, DOPAC and HVA in the perfusates were measured by high performance liquid chromatography (HPLC) with electrochemical detection. The basal level of DA was 2.76 +/- 0.64 pg/min, whereas the levels of DOPAC and HVA were 218.7 +/- 20.7 and 142.4 +/- 10.6 pg/min, respectively. Apomorphine (4 mg/kg, i.v.) reduced the efflux of DA and its metabolites. Haloperidol (0.4 mg/kg, i.v.) did not change DA release and produced only a minor increase of its metabolites. This increase of metabolites was inhibited by pargyline. Met-enkephalin (10(-4) M), substance P (10(-4) M) and acetylcholine chloride (10(-4) M) added to the perfusing medium increased the release of DA. Met-enkephalin also increased the release of DOPAC. gamma-Amino-n-butyric acid (GABA, 10(-4) M) reduced the release of DOPAC and HVA when added to the perfusing medium. Thyrotropin releasing hormone (TRH, 5 mg/kg, i.v.) increased the release of HVA. These findings indicated that different mechanisms mediated effects of neurotransmitters or drugs on the release and metabolism of DA in the rat striatum.

Studies on the association of 2-thiazolidinecarboxylic acid and antimony potassium tartrate: chiral recognition and prediction of absolute configuration by electrospray ionization mass spectrometry.

Optically active 2-thiazolidinecarboxylic acid (2-THC), a substrate for D-amino acid oxidase in animal kidney, is known to undergo racemization quickly in solution. The association of (+)- and (-)-2-THC with antimony potassium tartrate K(2)[Sb(2)(L or D-tart)(2)] was studied by electrospray ionization mass spectrometry (ESI-MS). We observed that relative intensities of associated ions in acetonitrile/water solution were changing as the racemization progressed. For [Sb(2)(L-tart)(2)](2-), the intensities of the associated ions increased as (+)-2-THC underwent racemization to a (-)-isomer; on the other hand, the intensity of the associated ion decreased as (-)-2-THC underwent racemization to a (+)-isomer. In the case of [Sb(2)(D-tart)(2)](2-), an opposite effect on the intensities of the associated ions was observed. The change in the intensities of associated ions can be used for chiral recognition of (+)-2-THC and (-)-2THC. Stereochemical models of the association of the optical isomers with [Sb(2)(L- or D-tart)(2)](2-) were constructed from the consideration of both hydrogen bonding of NH-O functions and HSAB (hard and soft acids and bases) interaction of S and Sb atoms. Comparison of the stereochemical models with the ESI-MS results enabled us to predict the absolute configurations of the 2-THC isomers.

Chiral recognition in association between antimony potassium tartrate and bis(L-alaninate)ethylenediamine cobalt(III) complexes using electrospray ionization mass spectrometry

The chiral recognition of metal complexes by a quick and sensitive mass spectrometric analysis was investigated. The principle is introduction of an external chiral standard compound and detection of the differential association with two optical isomers. Using electrospray ionization mass spectrometry we detected weak intermolecular association between the external chiral anion bis(mu-L-, D-tartrato)-diantimonate(III), [Sb2(L-, D-tart)2]2- and isomeric bis(L-alaninate) ethylenediamine cobalt(III) complex ions, [Co(L-ala)2(en)]+ in acetonitrile/water solution. The difference in the association with optical isomers of the Co complex was measured. The results were interpreted based on a model of intermolecular interaction involving hydrogen bonding. The prospects of the mass spectrometry method for chiral recognition using the external chiral negative ion [Sb2(L-, D-tart)2]2- was discussed.

Intramolecular electron-transfer reaction within a diprotein complex of cytochrome c with ferrylmyoglobin modified with diethylenetriaminepentaacetic acid.

Horse heart metmyoglobins modified with diethylenetriaminepentaacetic acid, metMb(DTPA)n (n=1, 2, 4, and 5), were characterized by a MALDI-TOF mass spectrometry, amino-acid sequence analysis, and UV-Vis and CD spectroscopies. The DTPA-binding sites on metMb were Lys47, Lys50, Lys87, Lys145, and Lys147 for metMb(DTPA)5, Lys47, Lys87, Lys145, and Lys147 for metMb(DTPA)4, Lys87 and Lys145 for metMb(DTPA)2, and Lys87 for metMbDTPA, respectively. The modified metMb(DTPA)n showed cytochrome c peroxidase-like activity more efficiently than native metMb: metMb(DTPA)5>metMb(DTPA)4>metMb(DTPA)2> metMbDTPA approximately equals native metMb. The first-order rate constants for the reactions of ferrylMb(DTPA)n (n=2, 4, and 5) with reduced cytochrome c [cyt c(II)] were saturated with concentrations of cyt c(II), suggesting that the electron transfer (ET) occurs within a diprotein complex. The intramolecular ET rate constants in the diprotein complex increased with increasing the number of DTPA ions. The reactions of native ferrylMb and ferrylMbDTPA with cyt c(II) obeyed a second-order rate law. A possible ET mechanism is proposed; cyt c(II) binds the DTPA-linked anionic patch around Lys87, Lys145, and Lys147 region of ferrylMb(DTPA)n.

[Effects of ambroxol HCl on the guinea pig tracheal mucous secretion and the rat pulmonary surfactant secretion].

The effects of orally administered ambroxol HCl (ambroxol) on guinea pig tracheal mucous secretion and rat pulmonary surfactant secretion were investigated histologically and biochemically. Ambroxol significantly increased the number of active goblet cells in guinea pig tracheal epithelium and total mucopolysaccharide level. Moreover, ambroxol significantly increased the neutral mucopolysaccharide level and PAS-positive substance in the guinea pig tracheal submucosal glands. Ambroxol did not show a significant effect on the content of the total phosphatidylcholine in rat lung lavage fluid, while ambroxol significantly increased the ratio of disaturated phosphatidylcholine to total phosphatidylcholine. From these results, it is suggested that ambroxol increases both the tracheal mucous secretion, especially the neutral mucopolysaccharide, and pulmonary surfactant secretion and these effects reflect part of the expectorant mechanism of the drug.