Fundamental study of 99mTc nanocolloids for inflammation imaging

The biodistribution of nanometer-sized colloidal particles (nanocolloids, Solco-Basle Ltd.) was examined in six rabbits with turpentine-induced abscesses in the musculature of their hind legs. 99mTc labelled colloids with a mean particle size less than 100 nm were administered to each rabbit intravenously. The biodistribution of the tracer was studied with a gamma camera 1, 3, and 24 hours after injection. Prominent uptakes were demonstrated in the liver, spleen, bone marrow, kidneys, and urinary bladder. On the other hand, the abscess and surrounding inflammatory edema were depicted as unclearly margined faint hot spots. Subsequently, the rabbits were sacrificed and activity per gram tissues were counted with a well-type gamma counter. Although the results were almost in agreement with a gamma camera study, it was noticed the activity counts in the abscess were significantly higher than that of control muscles.     

Theoretical assessment of optical resolution enhancement and background fluorescence reduction by three-dimensional nonlinear structured illumination microscopy using stimulated emission depletion

Three-dimensional structured illumination microscopy (SIM) enlarges frequency cutoff laterally and axially by a factor of two, compared with conventional microscopy. However, its optical resolution is still fundamentally limited. It is necessary to introduce nonlinearity to enlarge frequency cutoff further. We propose three-dimensional nonlinear structured illumination microscopy based on stimulated emission depletion (STED) effect, which has a structured excitation pattern and a structured STED pattern, and both three-dimensional illumination patterns have the same lateral pitch and orientation. Theoretical analysis showed that nonlinearity induced by STED effect, which causes harmonics and contributes to enlarging frequency cutoff, depends on the phase difference between two structured illuminations and that the phase difference of π is the most efficient to increase nonlinearity. We also found that undesirable background fluorescence, which degenerates the contrast of structured pattern and limits the ability of SIM, can be reduced by our method. These results revealed that optical resolution improvement and background fluorescence reduction would be compatible. The feasibility study showed that our method will be realized with commercially available laser, having 3.5 times larger frequency cutoff compared with conventional microscopy.     

Dehydratase-specific probes for fatty acid and polyketide synthases

We targeted the development of a dehydratase (DH)-specific reactive probe that can facilitate detection, enrichment, and identification of DH enzymes in fatty acid synthases (FASs) and polyketide synthases (PKSs). The first reported mechanism-based inactivator, 3-decynoyl-N-acetylcysteamine (3-decynoyl-NAC), while active against the Escherichia coli β-hydroxydecanoyl thiol ester DH FabA, translates poorly to an activity-based probe because of nonspecific reactivity of the thioester moiety. Here we describe the design, synthesis, and utility of a DH-specific probe that contains a sulfonyl 3-alkyne reactive warhead engineered to avoid hydrolysis or nonenzymatic inactivation. When coupled with a fluorescent tag, this probe targets DH enzymes from recombinant type I and type II FAS and PKS enzyme systems and in whole proteomes. Activity studies, including FabA inactivation and antibiotic susceptibility, suggest that this sulfonyl 3-alkyne scaffold selectively targets a common DH mechanism. These studies indicate that the DH-specific mechanism-based probe can greatly accelerate both the functional characterization and molecular identification of virtually any type of FAS and PKS in complex proteomes.     

Synthesis and activity of a metabolite of (S)-6-amino-5-(6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxamido)-3-methyl-1-phenyl-2,4-(1H,3H)-pyrimidinedione (CX-659S)

CX-659S (1) [(S)-6-amino-5-(6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxamido)-3-methyl-1-phenyl-2,4-(1H,3H)-pyrimidinedione], has been developed as a new type anti-inflammatory agent for the treatment of dermatitis. The structure of a major metabolite of CX-659S was determined as (S)-6-amino-5-[2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxo-1,4-cyclohexadienyl)butanamide]-3-methyl-1-phenyl-2,4-(1H,3H)-pyrimidinedione (2) by direct comparison with the synthesized authentic compound. The anti-inflammatory activity of 2 was equipotent with that of 1 on the contact hypersensitivity reaction (CHR) induced by picryl chloride (PC) in mice, suggesting that compound 2 contributes, at least in part, to the anti-inflammatory activity of CX-659S.     

Superweakly interacting massive particle solutions to small scale structure problems

Collisionless, cold dark matter in the form of weakly interacting massive particles (WIMPs) is well motivated in particle physics, naturally yields the observed relic density, and successfully explains structure formation on large scales. On small scales, however, it predicts too much power, leading to cuspy halos, dense cores, and large numbers of subhalos, in apparent conflict with observations. We consider super-WIMP dark matter, produced with large velocity in late decays at times 10(5) - 10(8) s. As analyzed by Kaplinghat in a more general setting, we find that super-WIMPs have sufficiently large free-streaming lengths and low phase space densities to help resolve small scale structure problems while preserving all of the above-mentioned WIMP virtues.     

Application of high-resolution ESI and MALDI mass spectrometry to metabolite profiling of small interfering RNA duplex

We investigated the application of a high-resolution Orbitrap mass spectrometer equipped with an electrospray ionization (ESI) source and a matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) mass spectrometer to the metabolite profiling of a model small interfering RNA (siRNA) duplex TSR#34 and compared their functions and capabilities. TSR#34 duplex was incubated in human serum in vitro, and the duplex and its metabolites were then purified by ion exchange chromatography in order to remove the biological matrices. The fraction containing the siRNA duplex and its metabolites was collected and desalted and then subjected to high-performance liquid chromatography (HPLC) equipped with a reversed phase column. The siRNA and its metabolites were separated into single strands by elevated chromatographic temperature and analyzed using the ESI-Orbitrap or the MALDI-TOF mass spectrometer. Using this method, the 5' and/or 3' truncated metabolites of each strand were detected in the human serum samples. The ESI-Orbitrap mass spectrometer enabled differentiation between two possible RNA-based sequences, a monoisotopic molecular mass difference which was less than 2 Da, with an intrinsic mass resolving power. In-source decay (ISD) analysis using a MALDI-TOF mass spectrometer allowed the sequencing of the RNA metabolite with characteristic fragment ions, using 2,4-dihydroxyacetophenone (2,4-DHAP) as a matrix. The ESI-Orbitrap mass spectrometer provided the highest mass accuracy and the benefit of on-line coupling with HPLC for metabolite profiling. Meanwhile, the MALDI-TOF mass spectrometer, in combination with 2,4-DHAP, has the potential for the sequencing of RNA by ISD analysis. The combined use of these methods will be beneficial to characterize the metabolites of therapeutic siRNA compounds. Copyright ? 2012 John Wiley & Sons, Ltd.     

The alpha effectiveness of the dating ESR signal in barite

The alpha effectiveness was investigated for an ESR signal due to ${\text{SO}}_3^{?}$ in barite by comparing the dose responses of the signal for gamma irradiation and for He ion implantation with an energy of 4 MeV. The value was obtained to be 0.043 ± 0.018 for samples from Morocco and to be 0.0089 ± 0.0006 for Kanehira Mine. The dose response for He ion implantation for the latter sample better fits to a correlation line but the signal shape changes with gamma ray dose, indicating contribution from an unknown radiation sensitive signal, while the former sample shows the same signal shapes. A value of 0.043 ± 0.018 is adopted as the alpha effectiveness of ${\text{SO}}_3^{?}$ signal in barite, which is used for ESR dating of submarine hydrothermal activity.     

Photoexcitation of K-shell and L-shell hollow beryllium

We have observed K-shell and L-shell hollow beryllium atoms (2s(2)2p3s and 1s3s(2)3p) created by photoexcitation using synchrotron radiation. Resonance shapes were fitted to the Fano profile and the parameters were deduced. A Dirac-Fock calculation was performed to identify the configuration of the peaks and to predict other hollow atomic peaks. The results of the calculation were in good agreement with the experimental data. The comparison of the transition strength has revealed that the three-electron photoexcitation to the 1s3s(2)3p configuration is stronger than the two-electron photoexcitation to the 2s(2)2p3s configuration. This is attributed to the large overlap between the 2s orbital of the ground state (1s(2)2s(2)) with the orbital of the L-shell hollow state (1s3s(2)3p).     

Bazzanane sesquiterpenoids from the New Zealand liverwort Frullania falciloba

Two new bazzanane-type sesquiterpenoids have been isolated from the New Zealand liverwort Frullania falciloba. Their structures were confirmed by NMR and CD spectroscopy and chemical reactivity.     

Diterpenoids and aromatic compounds from the three New Zealand liverworts Jamesoniella kirkii, Balantiopsis rosea, and Radula species

Three new aromatics were isolated from the New Zealand liverwort Balantiopsis rosea. A new bibenzyl was isolated from an unidentified Radula species, together with known bibenzyls. Jamesoniella kirkii yielded three known ent-isopimarane and two ent-kaurane diterpenoids. Their structures were confirmed by NMR techniques, chemical reaction, and X-ray crystallographic analysis.