Soluble factor from tumor cells induces heme oxygenase-1 by a nitric oxide-independent mechanism in murine peritoneal macrophages

Tumor target-derived soluble secretary factor has been known to influence macrophage activation to induce nitric oxide (NO) production. Since heme oxygenase-1 (HO-1) is induced by a variety of conditions associated with oxidative stress, we questioned whether soluble factor from tumor cells induces HO-1 through NO-dependent mechanism in macrophages. We designated this factor as a tumor-derived macrophage-activating factor (TMAF), because of its ability to activate macrophages to induce iNOS. Although TMAF alone showed modest activity, TMAF in combination with IFN-gamma significantly induced iNOS expression and NO synthesis. Simultaneously, TMAF induced HO-1 and this induction was slightly augmented by IFN-gamma. Surprisingly, however, induction of HO-1 by TMAF was not inhibited by the treatment with the highly selective iNOS inhibitor, 1400 W, indicating that TMAF induces the HO-1 enzyme by a NO-independent mechanism. While rIFN-gamma alone induced iNOS, it had no effect on HO-1 induction by itself. Collectively, the current study reveals that soluble factor from tumor target cells induces HO-1 enzyme in macrophages. However, overall biological significance of this phenomenon remains to be determined.     

Pore morphologies in disordered nanoporous thin films

Materials with nanometer size heterogeneities are commonplace in the chemical and biological sciences (e.g, polymer blends, microemulsions, gels) and often exhibit complex morphologies. Although this morphology has a dramatic effect on the materials' properties, it is often difficult to accurately characterize. We describe a method, using small-angle X-ray scattering data, of generating representative three-dimensional morphologies of isotropic two-phase materials where the morphology is disordered, and we apply this to thin films containing nanometer sized pores with a range of porosities (4-44%). These representations provide a visualization of the pore morphology, give the pore size scale and extent of interconnection, and permit the determination of the transitions from closed pore to interconnected pores to bicontinuous morphology.     

Metabolism of eupatilin in rats using liquid chromatography/electrospray mass spectrometry

Eupatilin (5,7-dihydroxy-3',4',6-trimethoxy flavone) is an active ingredient of an ethanol extract of Artemisia asiatica (DA-9601) that is used in the treatment of gastritis. In vitro and in vivo metabolism of eupatilin in the rats has been studied by LC-electrospray mass spectrometry. Rat liver microsomal incubation of eupatilin in the presence of NADPH and UDPGA resulted in the formation of four metabolites (M1-M4). M1, M2, M3 and M4 were tentatively identified as 3'- or 4'-O-demethyl-eupatilin glucuronide, eupatilin glucuronide, 6-O-demethyleupatilin and 3'- or 4'-O-demethyl-eupatilin, respectively. Those metabolites from in vitro study were also characterized in bile, plasma or urine samples after an intravenous administration of eupatilin to rats. In rat bile, plasma and urine samples, eupatilin glucuronide (M2) was a major metabolite, whereas M3, M4 and M4 glucuronide (M1) were the minor metabolites.     

Monoclonal antibody-based enzyme-linked immunosorbent assay for the insecticide imidacloprid

An enzyme-linked immunosorbent assay (ELISA) was developed for the neonicotinoid insecticide imidacloprid, 1-[(6-chloro-3-pyridinyl)methyl]-N-nitro-2-imidazolidinimine using monoclonal antibodies (MAb). Three MAbs, designated as E6A6, E6F3 and H7F7, were raised from mice immunized with an imidacloprid hapten-ovalbumin conjugate. These MAbs performed similarly in indirect competition ELISA (icELISA), so one, E6F3, was selected for detailed study. The equilibrium constants (K_d) and association and dissociation rate constants (k_on, k_off) for five neonicotinoids and one imidacloprid metabolite to E6F3 were determined by kinetic exclusion fluoroimmunoassay (KinExA). Affinities (1/K_d) of E6F3 for acetamiprid and clothianidin were similar, but 50-fold weaker than that of imidacloprid. MAb E6F3 had no measurable affinity for the other neonicotinoids. The icELISA can tolerate up to 15% (v/v) acetone or 20% (v/v) methanol. Assay sensitivity was similar at pH 4-9, 1-10-fold concentration of PBS with or without 0.05% Tween 20, and incubation times of 30-180 min. The half-maximal inhibition and the limit of detection were approximately 0.8 and 0.1 μg/l of imidacloprid in icELISA, and 0.3 and 0.03 μg/l in direct competition ELISA (dcELISA), respectively. Analysis of imidacloprid-fortified water and cucumber samples by the icELISA showed average recoveries from 70 to 120%.     

A theoretical investigation into the conformational changes of dibenzo-p-dioxin, thianthrene, and selenanthrene

Theoretical ab initio calculations using the HF and B3LYP methods have been carried out to investigate the conformational differences of three cyclic rings, dibenzo-p-dioxin (DPD), thianthrene (THT), and selenanthrene (SET). The physical origin for the conformational preference of each molecule has been studied using the natural bond orbital (NBO) analysis. The NBO results indicate that DPD exists in a planar form due to strong electron delocalization caused by the specific orbital interaction, around the X atom. On the other hand, THT and SET exist as puckered forms with high inversion barriers due to less effective electron delocalization. The NBO analysis also shows that the conformational stabilization in DPD is caused by a more effective overlap of the orbitals, compared with the overlap of the orbitals in THT.     

Phagocytosis of serum- and IgG-opsonized zymosan particles induces apoptosis through superoxide but not nitric oxide in macrophage J774A.1

Phagocytosis of serum- and IgG-opsonized zymosan (SOZ and IOZ, respectively) particles into J774A.1 macrophages induced apoptosis of the cells, accompanied by the expression of p21(WAF1), one of cyclin-dependent protein kinase (CDK) inhibitors. Furthermore, phagocytosis of SOZ and IOZ particles into macophages induced superoxide formation. Tat-superoxide dismutase (SOD), which is readily transduced into the cells using Tat-domain, protected the cells from the apoptosis induced by phagocytosis of SOZ and IOZ particles. lipopolysaccharide (LPS) /interferon-gamma (IFN-gamma) also caused the apoptosis of the cells. However, Tat-SOD could not protect the cells from LPS/IFN-gamma induced apoptosis, suggesting that apoptosis mechanisms involved are different from each other. In the present study, we determined the amounts of nitric oxide (NO) produced by SOZ, IOZ, and LPS/IFN-gamma, and found that SOZ and IOZ did not induce the generation of NO in macrophages, whereas LPS/ IFN-gamma did. The apoptosis due to phagocytosis was accompanied with the release of cytochrome c from mitochondrial membrane to cytosolic fraction. Furthermore, SOZ and IOZ induced the cleavage of procasapase-3 (35 kDa) to give rise to an active caspase-3 (20 kDa), which was blocked by Tat- SOD but not by 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (PTIO), a scavenger of NO. On the other hand, LPS/IFN-gamma caused the activation of procaspase-3, which was blocked by PTIO but not by Tat-SOD. Taken together, phagocytosis of SOZ and IOZ particles induced apoptosis through superoxide but not NO in macrophages, accompanied with the release of cytochrome c and the activation of caspase-3.     

Basicity of a stable carbene, 1,3-di-tert-butylimidazol-2-ylidene,in THF

The basicity of 1,3-di-tert-butylimidazol-2-ylidene (1) was measured in THF against three hydrocarbon indicators. Both ion pairs and free ions were found and the corresponding equilibrium constants were measured. Homoconjugation was not found in either THF or DMSO. The carbene is effectively more basic in DMSO by several pK units, probably because of hydrogen bonding of 1-H(+) to DMSO. Model ab initio computations are consistent with these results.     

Roles of terminal groups of oligomer electrolytes in determining photovoltaic performances of dye-sensitized solar cells

The effect of terminal groups of oligomer electrolytes on the photovoltaic performance of dye-sensitized solar cells (DSSCs) have been systematically investigated to show that the terminal group plays a critical role in determining the concentration of I(3)(-), ionic conductivity, flatband potential and consequently the energy conversion efficiency.