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Nicotinamide adenine dinucleotide phosphate oxidases (NOXs) are the major enzymatic source of reactive oxygen species (ROS). NOX2 and NOX4 are expressed in the heart but its role in hypoxia-induced atrial natriuretic peptide (ANP) secretion is unclear. This study investigated the effect of NOX on ANP secretion induced by hypoxia in isolated beating rat atria. The results showed that hypoxia significantly upregulated NOX4 but not NOX2 expression, which was completely abolished by endothelin-1 (ET-1) type A and B receptor antagonists BQ123 (0.3 µM) and BQ788 (0.3 µM). ET-1-upregulated NOX4 expression was also blocked by antagonists of secreted phospholipase A2 (sPLA2; varespladib, 5.0 µM) and cytosolic PLA2 (cPLA2; CAY10650, 120.0 nM), and ET-1-induced cPLA2 expression was inhibited by varespladib under normoxia. Moreover, hypoxia-increased ANP secretion was evidently attenuated by the NOX4 antagonist GLX351322 (35.0 µM) and inhibitor of ROS N-Acetyl-D-cysteine (NAC, 15.0 mM), and hypoxia-increased production of ROS was blocked by GLX351322. In addition, hypoxia markedly upregulated Src expression, which was blocked by ET receptors, NOX4, and ROS antagonists. ET-1-increased Src expression was also inhibited by NAC under normoxia. Furthermore, hypoxia-activated extracellular signal-regulated kinase 1/2 (ERK1/2) and protein kinase B (Akt) were completely abolished by Src inhibitor 1 (1.0 µM), and hypoxia-increased GATA4 was inhibited by the ERK1/2 and Akt antagonists PD98059 (10.0 µM) and LY294002 (10.0 µM), respectively. However, hypoxia-induced ANP secretion was substantially inhibited by Src inhibitor. These results indicate that NOX4/Src modulated by ET-1 regulates ANP secretion by activating ERK1/2 and Akt/GATA4 signaling in isolated beating rat hypoxic atria.

When attacked by herbivores, plants emit volatiles to attract parasitoids and predators of herbivores. However, our understanding of the effect of plant volatiles on the subsequent behaviour of conspecific parasitoids when herbivores on plants are parasitized is limited. In this study, rice plants were infested with gravid females of the brown planthopper (BPH) Nilaparvata lugens for 24 hr followed by another 24 hr in which the BPH eggs on plants were permitted to be parasitized by their egg parasitoid, Anagrus nilaparvatae; volatiles from rice plants that underwent such treatment were less attractive to subsequent conspecific parasitoids compared to the volatiles from plants infested with gravid BPH females alone. Chemical analysis revealed that levels of JA and JA-Ile as well as of four volatile compounds-linalool, MeSA, α-zingiberene and an unknown compound-from plants infested with BPH and parasitized by wasps were significantly higher than levels of these compounds from BPH-infested plants. Laboratory and field bioassays revealed that one of the four increased chemicals-α-zingiberene-reduced the plant's attractiveness to the parasitoid. These results suggest that host plants can fine-tune their volatiles to help egg parasitoids distinguish host habitats with parasitized hosts from those without.

Graphite oxide (GO), often being used as a precursor of graphene-based materials, can be prepared through a modified Hummers' method. The last step of GO synthesis, i.e. the removal of hydrogen chloride (HCl), is generally time-consuming, costly and polluting. Herein, the HCl-containing graphite oxide (H-GO) and pure GO aqueous suspensions were respectively hydrothermally treated followed by freeze-drying, leading to two graphene aerogels (H-GA and GA). Characterization results show that the two aerogels possess similar microstructure but distinct surface chemistry properties. When used as the active electrode materials in supercapacitors, GA and H-GA show comparable electrochemical properties in KOH aqueous electrolyte. In EMIMBF4 ionic liquid electrolyte, H-GA shows larger specific capacitance (243 F g−1 at 1 A g−1), better rate capability, and higher energy density (135 Wh kg−1 at 1 kW kg−1) compared with GA (216 F g−1, 120 Wh kg−1), which is attributed to its higher O/C ratio and better electrolyte wettability. We propose that it is not always necessary to conduct the HCl removal, and H-GO could be directly used as a precursor, which not only saves the time and cost remarkably but also may open up a relatively facile route to fabricate some graphene-based functional bulk materials.

We demonstrate a proof-of-concept design of a new platform for proton recognition and modulation. The new proton receptors are derived from a unique class of synthetic supercontainers that exhibit exceptional proton binding capacity (over 50 equiv.) and intriguing proton-dependent fluorescent switching behavior. Experimental and computational studies suggest that the proton-responsive event involves a two-step mechanism pertaining to proton binding by both amino and pyrenyl moieties of the supercontainer constructs. The high proton binding capacity of the supercontainers can be further modulated via small-molecule "regulators" that compete for the proton-binding sites, opening exiting new opportunities for proton manipulation in both chemistry and biology.

In this paper, we developed a low-cost and easy-to-fabricate hydrogel-based localized surface plasmon resonance (LSPR) substrate for sensitive, consistent, specific, and time-lapse detection of hydrogen peroxide (H2O2) secreted from living cells in cell culture medium. The uniform and reproducible LSPR substrate was made from agarose, gold nanorods (AuNRs), horseradish peroxidase (HRP), and 3-amino-9-ethylcarbazole (AEC). In H2O2 sensing, insoluble precipitates were generated in the gel film by an HRP-catalyzed reaction in the presence H2O2, and the precipitates near the AuNRs caused a LSPR shift of the substrate, which could be used for H2O2 quantification. Using the developed substrate, we showed that a H2O2 concentration as low as 0.5 μM could be detected in complete cell culture medium within 30 min with good reproducibility. In addition, we demonstrated time-lapse detection of H2O2 secreted from living cells after the cells were stimulated by ascorbic acid. The results indicated that the agarose gel of the substrate provided the filtration capability and the improved stability of the AuNRs allowing detection of H2O2 in cell culture medium without tedious sample preparation procedures. Moreover, the HRP-catalyzed reaction-based LSPR biosensing offered good selectivity for H2O2 detection and possessed great potential for various in vitro cell studies.

The discovery of novel tetrahydropyrrolo[1,2-c]pyrimidines derivatives from Bay41_4109 as hepatitis B virus (HBV) inhibitors is herein reported. The structure–activity relationship optimization led to one highly efficacious compound 28a (IC50 = 10 nM) with good PK profiles and the favorite L/P ratio. The hydrodynamic injection model in mice clearly demonstrated the efficacy of 28a against HBV replication.

Aims Gallic acid (GA) protects against myocardial ischemia-reperfusion (I/R) injury, prevents cardiac hypertrophy and fibrosis, and has anti-inflammatory activity in the heart. However, its effects in regulating atrial natriuretic peptide (ANP) secretion are unknown. The aim of this study was to determine the function of GA in regulating ANP secretion and atrial dynamics in rat atria. Key findings GA (0.01, 0.05, and 0.1 μmol/L) significantly increased atrial ANP secretion and induced positive inotropy dose-dependently. GA (0.1 μmol/L) also increased plasma level of ANP and hemodynamics in rats. These effects were accompanied by upregulation of atrial protein kinase C subtypes β and e (PKC β and PKC e ), which was completely blocked by LY333531 and EAVSLKPT, antagonists of protein PKC β and PKC e , respectively. GA-induced ANP secretion was also attenuated by Go6983 but not rottlerin, antagonists of PKC α and PKC δ , and the positive inotropy was reversed by Go6983. U-73122, a phospholipase C (PLC) antagonist, mitigated the effects of GA on ANP secretion and mechanical dynamics and downregulated Phospho-PLC β at Ser 537 (pPLC β S537), Phospho-PLC β at Ser 1105 (pPLC β S1105), PKC β and PKC e levels, whereas KN62, an inhibitor of Ca 2 + /calmodulin-dependent kinase II, was not modified the GA-induced ANP secretion and suppressed GA-induced mechanical dynamics. Significance GA promotes ANP secretion and effects positive inotropy with regard to mechanical dynamics through the activation of PLC-PKC signaling in rat atria.

A rasagiline transdermal patch can be used to offer continuous rasagiline to patients with Parkinson's disease who cannot take their usual oral medications. This was the first study to investigate the pharmacokinetics, pharmacodynamics, and safety of the rasagiline transdermal patch in healthy Chinese subjects. Thirty subjects were randomized to 3 groups with 10 subjects in each group. The 10 subjects of group 1 received a single 1-mg dose of rasagiline as a tablet; the 20 subjects of groups 2 and 3 received a single transdermal patch (48-hour patch-on period) containing 1.25 mg and 2.5 mg rasagiline, respectively. After a 2-week washout period, the subjects of group 1 were assigned to receive 1 mg of rasagiline tablets every 24 hours for 7 days, and the subjects of group 2 were assigned to receive 1.25-mg rasagiline transdermal patches (48-hour patch-on period) every 72 hours for 5 time periods. The absorption of rasagiline from the transdermal patch was significantly improved, although the peak plasma concentration was obviously reduced. There was slight accumulation of rasagiline dose after multiple administrations. Inhibition of platelet monoamine oxidase-B (MAO-B) activity was dose dependent. The 80% inhibition maintained for at least 48 hours after multiple-dose administration of 1 mg tablets, and for 72 hours after multiple-dose administration of 1.25 mg/48 h patch. Compared with rasagiline tablets, the transdermal patch had a prolonged duration of 80% inhibition and increased maximal inhibition of MAO-B activity. These characteristics permitted an interval of 3 days of dosing, which was convenient for patients to use.

Pregnant rats were treated intraperitoneally with a single dose of methotrexate (MTX) 90 mg/kg on gestation day (GD) 13, and fetal eyeballs were examined time-dependently from GD 13.5 to 15.5. Throughout the experimental period, the inner plate of the ocular cup in the MTX group was significantly thinner than that in the control group. In the inner plate of the ocular cup on GD 15 and 15.5, whereas a developed ganglion cell layer was observed in the control group, the ganglion cell layer in the MTX group was undeveloped and indistinguishable. Disturbance of the arrangement of lens fiber cells, narrowing of the hyaloid cavity of the optic cup, and hypoplasia of optic nerve fibers were observed in the MTX group on GD 15 and 15.5. Increase of pyknosis and decrease of mitosis were induced in the optic cup and the lens epithelium of the MTX group. In the inner plate of the optic cup and the lens epithelium of the MTX group, the cleaved caspase-3- and TUNEL-positive rates increased significantly throughout the experimental period. The phospho-histone H3-positive rate in the inner plate of the optic cup decreased significantly from GD 13.5 to 14.5, and it recovered on GD 15. On the other hand, the phospho-histone H3-positive rate in the lens epithelium decreased significantly throughout the experimental period. These results suggested that optic tissue on GD 13 in rats was sensitive to MTX.

A new sulfonylcalix[4]arene-based dinuclear terbium molecular container (1) was conveniently synthesized and utilized as a fluorescence probe for the detection of a well-known biomarker 1-hydroxypyrene (1-OHP), which is used for the evaluation of polycyclic aromatic hydrocarbons (PAHs). The sulfonylcalix[4]arene ligand could not only serve as an efficient antenna ligand to promote the ligand-to-metal energy transfer but also provide a suitable cavity to accommodate 1-OHP. Promising fluorescence quenching effects were well established during the titration of the compound 1 with 1-OHP, and these effects were due to the enhancement in the host–guest intermolecular charge transfer and the decrease in the ligand-to-metal energy transfer after the formation of the stable host–guest complex. The fluorescence sensing mechanism was clearly understood through the titration experiments, and the data could be fit with the Benesi–Hildebrand and Stern–Volmer models. The TbIII-TBSC-based luminescent sensor exhibited quick response, high sensitivity, and specific selectivity to 1-OHP, even in the presence of other constituents in urine, thus providing a new sensing platform for the clinical diagnosis of human exposure to PAHs.

A novel, Gram-staining-negative, rod-shaped, aerobic and motile bacterium, designated strain CC-SKC2(T), was isolated from the root tumor of a green bell pepper (Capsicum annuum var. grossum) plant in Taiwan. Cells were positive for oxidase and catalase activities and exhibited growth at 25-37 °C, pH 4.0-9.0 and tolerated NaCl concentrations up to 4.0 % (w/v). Strain CC-SKC2(T) is able to trigger nodulation in soybean (Glycine max Merr.), but not in Capsicum annuum var. grossum, red bean (Vigna angularis), sesbania (Sesbania roxburghii Merr.) or alfalfa (Medicago varia Martin.). The novel strain shared highest 16S rRNA gene sequence similarity to Rhizobium rhizoryzae KCTC 23652(T) and Rhizobium straminoryzae CC-LY845(T) (both 97.5 %) followed by Rhizobium lemnae L6-16(T) (97.3 %), Rhizobium pseudoryzae KCTC 23294(T) (97.1 %), and Rhizobium paknamense NBRC 109338(T) (97.0 %), whereas other Rhizobium species shared96.7 % similarity. The DNA-DNA relatedness values of strain CC-SKC2(T) with R. rhizoryzae KCTC 23652(T), R. pseudoryzae KCTC 23294(T) and R. paknamense NBRC 109338(T) were 11.4, 17.2 and 17.0 %, respectively (reciprocal values were 11.1, 28.3 and 24.0 %, respectively). Phylogenetic analysis based on 16S rRNA, atpD and recA genes revealed a distinct taxonomic position attained by strain CC-SKC2(T) with respect to other Rhizobium species. The major fatty acids in strain CC-SKC2(T) were C16:0, C19:0 cyclo ω8c, C14:0 3-OH and/or C16:1 iso I and C18:1 ω7c and/or C18:1 ω6c. The polyamine pattern showed predominance of spermidine and moderate amounts of sym-homospermidine. The predominant quinone system was ubiquinone (Q-10) and the DNA G+C content was 60.5 mol%. On the basis of polyphasic taxonomic evidence presented here, strain CC-SKC2(T) is proposed to represent a novel species within the genus Rhizobium, for which the name Rhizobium capsici sp. nov. is proposed. The type strain is CC-SKC2(T) (=BCRC 80699(T) = JCM 19535(T)).

Color tunable electroluminescence is obtained from devices having a spin-coated single emitting layer with a novel EuIII-tetrakis-β-diketonate complex NBu4[EuL4] (L− = 4-(4′-carbazol-9-yl-biphenyl-4-yl)-1,1,1-trifluoro-4-oxo-but-2-en-2-ol anion) doped with CBP (4,4′-N,N′-dicarbazolebiphenyl)–OXD7 (1,3-bis[2-(4-tert-butylphenyl)-1,3,4-oxadiazo-5-yl]benzene) blend. An external quantum efficiency (EQE) as high as 2.8%, maximum brightness (L) of 1547 cd m−2, current efficiency (ηc) of 5.14 cd A−1 and power efficiency (ηp) of 2.53 lm W−1 were shown by a device having the structure ITO/PEDOT:PSS (50 nm)/CBP (56%):OXD7 (24%):NBu4[EuL4] (20%) (50 nm)/TPBI (50 nm)/LiF (1.5 nm)/Al (120 nm). The newly designed β-diketonate ligand containing hole-transporting carbazole group and π-spacer extends the excitation window of the EuIII-complex to 450 nm and also improves the carrier transport properties. The modification in the ligands results in a low turn-on voltage of the devices (

Electrical discharge grinding is part of the most widely used methods to machine polycrystalline cubic boron nitride cutting tool. Polycrystalline cubic boron nitride compact samples processed in domestic with different grain size are the research object. Electrode running speed, peak current, and pulse width are selected as the main process parameters. The material removal volume and electrode loss set as the evaluation index of productive efficiency; workpiece surface roughness value sets as an evaluation standard of processing quality. Through electrical discharge grinding experiments, combined with scanning electron microscopy observation, energy spectrum analyzer and roughness tester, the influences of the main process parameters on electrical discharge grinding are analyzed, polycrystalline cubic boron nitride compact electrical discharge grinding technics & mechanism are summarized.

Lipocalin-type prostaglandin D synthase (L-PGDS) and peroxisome proliferator activated receptor γ (PPARγ) play important roles in cardiovascular diseases. Nevertheless, effects of hypoxia-inducible factor 1α (HIF-1α) on L-PGDS and PPARγ protein levels and its role in hypoxia-induced atrial natriuretic peptide (ANP) secretion are unclear. In perfused beating rat atria, we observed that hypoxia significantly increased HIF-1α protein levels and stimulated ANP secretion, while upregulating L-PGDS. Hypoxia-induced ANP secretion was clearly attenuated by HIF-1α antagonist 2-methoxyestradiol, downregulating both HIF-1α and L-PGDS protein levels. It was also attenuated by L-PGDS antagonists, AT-56 and HQL-49, downregulating L-PGDS protein levels. In addition, hypoxia-induced ANP secretion was accompanied by increased PPARγ protein levels and was strongly attenuated by PPARγ antagonist GW9662. Hypoxia-induced increase in atrial PPARγ protein levels were dramatically inhibited by both 2-methoxyestradiol and AT-56. These results indicated that hypoxia promotes ANP secretion, at least in part, by activating HIF-1α-l-PGDS-PPARγ signaling in beating rat atria.

High speed turn-milling has superiority on the productivity and the quality of work pieces, and is more suitable to machine micro-shaft parts and desirable miniature parts based on the turn-milling technology. In this papers adopting orthogonal experiment method cutting experiments of orthogonal turn-milling Aluminum alloy have been done. The relation between turn-milling regimes (cutter rotate speed, axial feed, feed per tooth etc.) and machined surface roughness has been ascertained. Finally, primary and secondary order of cutting regimes impacting surface roughness has more been confirmed through orthogonal experiments variance analysis, the rotate speed of cutter (cutting speed) influence greatly on surface roughness. Through 2-dimension surface topography diagram and 3-dimension surface topography of processed surface, it can be seen that high speed turn-milling processing technology can process micro miniature component of high surface quality, and features excellent development prospect and application value.

In this work, the thermal bistability of magnons in yttrium iron garnet microspheres was experimentally characterized. Using high quality factor optical whispering gallery modes in the microsphere, the temperature change caused by the microwave pump is precisely measured by probing the optical resonance wavelength shift. It is found that temperature change also induces the magnonic frequency shift and the frequency shift depends on the crystal axis direction of the material. With some appropriate alignment of the magnetic field and the microsphere, the magnonic frequency shift induced by the thermal bistability from the microwave can be suppressed. Our results show its potential use in future experiments on magnon-based transducers.

The adsorption of phenol with the crosslinked polymer of P(MMA-MAh)-PEG33 was studied. Through the research, it was found the adsorption was favored in acidic aqueous solution and almost needed 35 h to reach the equilibrium. The calculated thermodynamic parameters suggested that the adsorption of phenol onto P(MMA-MAh)-PEG33 was spontaneous and exothermic in nature. From the kinetic research, the adsorption of phenol with the crosslinked polymer belonged to the first-order reaction. The adsorption isotherms of phenol with polymer well accorded with the Freundlich model.

Controlled drug delivery systems employing microparticles offer lots of advantages over conventional drug dosage formulations. Microencapsulation technique have been conducted with biodegradable polymers such as poly(lactic-co-glycolic acid) (PLGA) and poly(lactic acid) (PLA) for its adjustable biodegradability and biocompatibility. In this study, we evaluated two techniques, oil-in-water (o/w) emulsion solvent evaporation and spray drying, for preparation of polymeric microparticles encapsulating a newly synthesized drug, SS-AG20, for the long-term drug delivery of this low-molecular-weight drug with a very short half-life. Drug-loaded microparticles prepared by the solvent evaporation method showed a smoother morphology; however, relatively poor encapsulation efficiency and drastic initial burst were discovered as drawbacks. Spray-dried drug-loaded microparticles had an imperfect surface with pores and distorted portions so that its initial burst was critical (70.05-87.16%) when the preparation was carried out with a 5% polymeric solution. By increasing the concentration of the polymer, the morphology was refined and undesirable initial burst was circumvented (burst was reduced to 35.93-74.85%) while retaining high encapsulation efficiency. Moreover, by encapsulating the drug with various biodegradable polymers using the spray drying method, gradual and sustained drug release, for up to 2 weeks, was achieved.

Tisplatin is one of the valuable icancer agents against several types of neoplasm. However, nephrotoxicity is the major adverse effect representing in cisplatin therapy. In this study, the animal tests detecting protective effects of a natural compound, Decursin, on cisplatin-induced nephrotoxicity were examined by using in vivo model. Pretreatment Decursin 10, 20 and 40 mg · kg −1 at 48, 24 and 6 h, and administration of a single dose of Cisplatin 5.2 mg · kg −1 . Nephrotoxicity was evaluated by serum BUN and creatinine examination. There was significant difference in body weights, serum BUN and creatinine levels of the normal group. Based on the new understanding of the protective mechanisms of cisplatin-induced nephrotocivity, new strategies can be developed to prevent renal injury or to enhance recovery after cisplatin treatment.

Hemodialysis vascular access dysfunction (HVAD) due to the aggressive development of venous neointimal hyperplasia remains a major complication for patients with synthetic arteriovenous grafts. Paclitaxel-coated expanded polytetrafluoroethylene (ePTFE) grafts effectively prevent neointimal hyperplasia and stenosis. However, peri graft inflammation or edema can be another complication of ePTFE grafts, preventing early cannulation. Three different types of ePTFE grafts, including grafts without paclitaxel coating (control group, n = 12), grafts with paclitaxel coating at a dose density of 0.61 ug/mm 2 (low concentration group, n =12), and grafts with paclitaxel coating at a dose density of 1.15 ug/mm 2 (high concentration group, n = 12) were placed in the backs of 12 rabbits, simultaneously. Six rabbits were euthanized after one week and the remaining six were euthanized two weeks after implantation. Perigraft inflammation, graft wall inflammation, stromal cell proliferation, blood vessel formation, tissue necrosis and edema were analyzed for the grafts in each animal. Inflammation surrounding the paclitaxel-coated grafts was significantly reduced compared to the control group. Stromal cell layers were detected at the interface between the graft and the surrounding tissue in the control group, infiltrated into the graft interstices, and differentiated into myofibroblasts for graft healing. Paclitaxel-coated grafts inhibited stromal cell proliferation and infiltration into the graft wall. Tissue necrosis and edema were not detected in either of the paclitaxel-coated graft groups.

We synthesized biodegradable b-PEIS (branched poly(ethylenimine sulfide)) by crosslinking linear PEIS. We controlled the degree of crosslinking and molecular weight by adjusting the amount of the crosslinker, bisepoxide. The b-PEIS was readily degradable under reductive conditions (5mm glutathione solution) and the degradation time was dependent on the degree of crosslinking. We controlled the molecular weights of the b-PEIS by regulating the amount of crosslinker and thus, the degree of crosslinking. Our titration data showed that there was almost no loss in buffering ability before or after bisepoxide crosslinking. We verified the degradation of this polymer by MALLS and gel electrophoresis, and confirmed that there was a high transfection efficiency and low cytotoxicity based on cellular data. Intracellular trafficking was observed by image restoration microscopy, demonstrating that b-PEIS does not accumulate in the cell interior.

The characteristics of the thermal evolution of aromatic maturity parameters, such as methyl phenanthrene, alkylated dibenzothiophene, dehydroxy-tocopherol and triaromatic-sterane series compounds are discussed on the basis of molecular organic geochemistry and GC-MS analysis of the source rocks in the Dongling area of the Songliao Basin. The distribution characteristics of aromatic hydrocarbon chromatography, the methyl phenanthrene index (MPI), the ratio of β/γ dehydroxy-tocopherol, 4-/1-MDBT, 4,6-/1,4-DMDBT, TASC27 (20R)/C28 (20R), the content of the triaromatic-sterane series and vitrinite reflectance measurements all suggest that the Lower Cretaceous Quantou Formation source rock is in the middle stage of maturity, whereas the Lower Cretaceous Yingcheng Formation and the Upper Jurassic Huoshiling Formation source rocks have reached a high maturity stage.

Protein phosphatase-1 (PP1) is one of the most important Ser/Thr phosphatases in eukaryotic cells. G274, E275, and F276 are located at the tip of the beta12-beta13 loop of protein phosphatase-1. Without Mn2+, the basal activities and intrinsic fluorescence spectra of all single and double deletion mutants of G274, E275, and F276 were similar to those of PP1, but deletion mutants DeltaE275 and DeltaE275F276 showed hyperactivity and corresponding changes in intrinsic fluorescence spectra when Mn2+ was present. This suggests that the conformation transition resulting from the combined effect of mutation and Mn2+ accounts for the hyperactivity of mutants. These observations imply that E275 and F276 play a role in resisting enzyme activation by Mn2+ in PP1.

Protein phosphatase 1 consists of a secondary structure arrangement, conserved in the serine/threonine protein phosphatase gene family, flanked by nonconserved N-terminal and C-terminal domains. The deletion mutant of PP1 with the 8 nonconserved N-terminal residues removed was designated PP1-(9-330). PP1-(9-330) had a higher activity and affinity than PP1 when assayed against four different substrates, and it also demonstrated a 6-fold higher sensitivity to the inhibitor okadaic acid. This suggested that the N-terminal domain suppressed the activity of PP1 and interfered with its inhibition by okadaic acid. The ANS fluorescence intensity of PP1-(9-330) was greater than that of PP1, which implies that the hydrophobic groove running from active site in the truncated PP1 was more hydrophobic than in PP1. Our findings provide evidence that the nonconserved N-terminus of PP1 functions as an important regulatory domain that influences the active site and its pertinent properties.

Surface treatment of glass bead (GB) was carried out by using γ-glycidoxypropyltrimethoxy silane (GPTES) and γ-methacryloxypropyltrimethoxy silane (MPTMS) as coupling agents, respectively. The steady viscosity and yield stress of the GB/hydroxyl terminated polybutadiene (HTPB) suspensions were determined by Brookfield R/S rheometer. The effect of surface treatment on the viscosity and yield stress of GB/HTPB suspension was investigated. The results indicate that the viscosity of the pristine GB/HTPB suspension increases with increasing GB, and the relationship between its viscosity and volume fraction of GB depends on the shear rate. The modification of GB by MPTMS changes the viscosity of the MPTMS@GB/HTPB suspension, and its viscosity is the minimum at the MPTMS dosage of 0.3 g per 1 g GB. Additionally, the modification of GB by MPTMS increases the yield stress of the GB/HTPB suspension, and its yield stress is the maximum at the MPTMS dosage of 0.1 g per 1 g GB. The GPTES modified GB/HTPB suspension behaves lower viscosity and weaker shear thinning than the MPTMS modified GB/HTPB suspension within the range of experimental shear rate.

We designed chiral 2-nitroimidazole derivatives containing a 2-aminomethylene-4-cyclopentene-1,3-dione moiety as antiangiogenic hypoxic cell radiosensitizers. Based on results of molecular orbital calculations, the 2-aminomethylene-4-cyclopentene-1,3-dione moiety is expected to show high electrophilicity comparable to that of the 2-methylene-4-cyclopentene-1,3-dione moiety included in TX-1123 and tyrphostin AG17. We evaluated the antiangiogenic and radiosensitizing effects of the new compounds, along with other biological properties including their activities as hypoxic cytotoxicities and protein tyrosine kinase (PTK) inhibitory activities. Among the compounds tested, 5 (TX-2036) proved to be the strongest antiangiogenic hypoxic cell radiosensitizer. All the other chiral 2-nitroimidazole derivatives having 2-aminomethylene-4-cyclopentene-1,3-dione moiety tested were also antiangiogenic hypoxic cell radiosensitizers. The PTK inhibitory activity of 5 (TX-2036) showed this to be a promising and potent EGFR kinase inhibitor, having an IC(50) value of lower than 2microM. This compound also was an Flt-1 kinase inhibitor having an IC(50) value of lower than 20microM. Our results show that these chiral 2-nitroimidazole derivatives that contain the 2-aminomethylene-4-cyclopentene-1,3-dione moiety as a potent antiangiogenic pharmacophoric descriptor are promising lead candidates for the development of antiangiogenic hypoxic cell radiosensitizers.

In this study, we investigated the in vitro anti-obesity, anti-cancer activity and single oral dose toxicity of Inonotus xeranticus extracted by methanol (INXM) or ethyl acetate (INXE). In order to investigate anti-obestity effect of Inonotus xeranticus extracts, the 3T3-L1 cells were treated with these extracts at various concentrations(1, 10, 100 and 300 ㎍/㎖). It was observed that 3T3-L1 cells treated with 100 ㎍/㎖ of Inonotus obliquus ethyl acetate extract (INOE), INXM and INXE, in the absence of differentiation cocktail (0.5 mM isobutylmethylxanthine (IBMX) 1 μM dexamethasone, 1 ㎍/㎖ insulin), differentiated at a rate of 78.5, 80.9, and 76.4% respectively. Differentiation rates of 86.6% and 83.4% were observed in 3T3-L1 cells which were treated with differentiation cocktail at 100 ㎍/㎖ of INXM and INXE, respectively. The anti-cancer effect of Inonotus xeranticus extracts was investigated using a method of sulforhodamine B in sarcoma 180 cell line. The cells were treated with these extracts (1, 10, 100 and 300 ㎍/㎖) for 48 hours. The growth of cells which were treated with 300 ㎍/㎖ of INXM was inhibited by 80.1%. The growth of sarcoma 180 cells which were treated with 100 and 300 ㎍/㎖ of INXE was inhibited by 74.7% and 64.5%, respectively. In single oral dose toxicity study, no differences were observed between control and treated groups in clinical signs, body weight gains, and feed and water consumptions. The results indicated that Inonotus xeranticus extracts did not show any toxic effects at 2,000 ㎎/㎏ in mice, and the LD?? of these extracts was found to be higher than 2,000 ㎎/㎏ in this experiment. From the above results, Inonotus xeranticus methanol and ethyl acetate extracts might have useful clinical applications in the management of cancer and obesity and may also be useful as a medicinal food.

1 H NMR. The newly synthesized polymer, PAMAM-R-PO was able to completely retard pDNA from a charge ratio of 2. The average diameter of PAMAM-R-PO polyplex was found to be 242 nm at a charge ratio of 30. The Zeta-potential value of PAMAM-R-PO polyplex was able to reach 20-30 mV over a charge ratio of 10. PAMAM-R-PO indicated higher cell viability than unmodified PAMAM-R on HeLa and 293 cells because of its hydroxylated amines. Transfection experiments on 293 cells showed that the transfection efficiency of PAMAM-R-PO was found to be 1.5-1.9 times higher than that of PEI25kDa at a charge ratio of 30. The polymer eventually displayed about 2 times greater transfection efficiency than PAMAM-R at the same charge ratio in the absence of serum. Therefore, we concluded that the modification of primary amines of PAMAMR to amino alcohols gives positive effects such as reduced cytotoxicity and enhanced transfection efficiency on 293 cells for gene delivery potency of PAMAM-R.

The purpose of this study is to evaluate the feasibility of human amniotic membrane (HAM) as a chondrocyte carrier by assessing cell proliferation and maintenance of phenotype in vitro and cartilage regeneration in vivo. Intact HAM was treated with 0.1% trypsin-ethylenediaminetetraacetic acid (EDTA) for 15 min and the epithelial cells removed to make a denuded HAM. Rabbit articular chondrocytes were then seeded on three different HAM substrates: the epithelial side of intact HAM (IHE), basement side of denuded HAM (DHB), and stromal side of denuded HAM (DHS). These cell-substrate specimens were cultured for up to 4 weeks, and cell proliferation rate and phenotypic stability were examined at weeks 1 and 4. While chondrocytes grew in monolayer fashion on the surface of IHE and DHB substrates, the cells seeded in DHS penetrated and spread into the whole thickness of the stromal layer. The proliferating activity of chondrocytes in DHB was continuously up-regulated. A similar proliferating activity was observed in DHS in the first week, which remained stable for up to 4 weeks. The expression of type II collagen gradually increased with time in the DHS group, while it gradually decreased in the DHB group or was not detected at all in the IHE group. These results suggested that denuded HAM was able to support chondrocyte proliferation and maintenance of phenotype in vitro, seemingly more favorable when DHS was used. Based on this data, the DHS with chondrocytes was used to cover rabbit osteochondral defect with the stromal side facing in. The defect area was successfully regenerated with hyaline cartilage in the Safranin-O stain and International Cartilage Repair Society (ICRS) scoring after 8 weeks of implantation. In conclusion, our findings suggest that denuded HAM could be one of the ideal cell carrier matrices for cartilage regeneration.

A Gram-stain-positive, rod-shaped, endospore-forming bacterium (designated strain CC-Bw-5T) was isolated from chopped tomato stems. The isolate grew at 20–40 °C, pH 6.0–8.0 and tolerated 6 % (w/v) NaCl. The most closely related strains in terms of 16S rRNA gene sequence similarity wereBacillus isabeliae(95.3 %) andBacillus oleronius(95.3 %). The G+C content of the genomic DNA was 37.2 ± 3.6 mol%. Strain CC-Bw-5Twas determined to possess C16 : 0, iso-C14 : 0, iso-C15 : 0, anteiso-C15 : 0and iso-C16 : 0as predominant fatty acids. The polar lipid profile consisted of predominant amounts of diphosphatidylglycerol, and moderate-to-trace amounts of phosphatidylethanolamine, phosphatidylglycerol, two unidentified glycolipids, an unidentified phospholipid and an unidentified phosphoglycolipid. The cell-wall peptidoglycan containedmeso-diaminopimelic acid; menaquinone (MK-7) was the predominant respiratory quinone. According to distinct phylogenetic, phenotypic and chemotaxonomic features, strain CC-Bw-5Tis proposed to represent a novel species within the genusBacillusfor which the nameBacillus lycopersicisp. nov. is proposed. The type strain is CC-Bw-5T( = BCRC 80623T = JCM 19140T).

A Gram-stain-positive, aerobic, non-motile, rod-shaped, yellow-pigment-producing bacterium, (designated strain CC-CZW004T), was isolated from seafood samples (sea urchins) at Penghu Island in Taiwan. Strain CC-CZW004Tgrew optimally at pH 7.0 and 30 °C in the presence of 3 % (w/v) NaCl. Phylogenetic analyses based on 16S rRNA genes revealed a distinct taxonomic position attained by strain CC-CZW004Twith respect to other members of the genusNocardioides. The novel strain shared highest 16S rRNA gene sequence similarities toNocardioides daejeonensisJCM 16922T(96.4 %),Nocardioides pacificusJCM 19260T(96.3 %), andMarmoricola scoriaeJCM 17444T(96.2 %). The major fatty acids of strain CC-CZW004Tconsisted of C17:0, C16:1ω5c, C17:1ω8c`, iso-C16:0and C19:1ω11c/C19:1ω9c(summed feature 6). The diagnostic diamino acid in the cell wall wasll-2,6-diaminopimelic acid. The polar lipid profile was composed of major amounts of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and aminophospholipid. The DNA G+C content was 73.3 mol%. The predominant quinone system was menaquinone (MK-8). On the basis of polyphasic taxonomic evidences presented here, strain CC-CZW004Trepresents a novel species of the genusNocardioides, for which the nameNocardioides echinoideorumsp. nov. is proposed. The type strain is CC-CZW004T( = BCRC 16974T = JCM 30276T).

Sixty-five new vibronic levels of the Na2 43g+ state have been observed in the 3390035200 cm1 energy region above the potential minimum of the ground state by pulsed perturbation facilitated opticaloptical double resonance (PFOODR) fluorescence excitation spectroscopy. These new data fill the gap between the low-v levels mainly observed by continuous wave (CW) PFOODR spectroscopy and the high-v levels above the 3s + 3d limit observed by pulsed PFOODR with predissociation detection. Molecular constants are fitted below potential shelf around the 3s + 3d atomic limit with previously published data (mainly observed by CW PFOODR) and these new data. RKR potential curve has been calculated with the new constants. The constants are: Te = 32127.090 cm1; e = 121.4099(0.20720) cm1; Be = 0.116287(0.0002300) cm1; Re = 3.551 A. An error of the RKR potential curve of J. Chem. Phys. 108, 7707 (1998) is corrected.

A Gram-stain negative, aerobic, rod-shaped and non-spore-forming bacterium, CC-Bw-6(T) was isolated from chopped tomato stems. Analysis of the 16S rRNA gene sequence of the strain CC-Bw-6(T) showed its affiliation with the genus Lysobacter within the class Gammaproteobacteria. Strain CC-Bw-6(T) was found to be most closely related to Lysobacter panaciterrae KCTC 12601(T) (97.0%) and Lysobacter daecheongensis KCTC 12600(T) (96.8%), and showed lower similarity (

High molecular weight (MW) hyaluronan (HA) preparation is considered to be more biologically active than HAs of lower MWs. However, many of the HA preparations currently used to treat osteoarthritis (OA) have lower MWs by the enhanced penetration of HA molecules into the synovial lining cells. In this study, we determined the effectiveness of sonophoresis on the delivery of high MW HA into synovial membrane using an animal model of OA. A total of 1000 kDa (HA1000) and 3000 kDa (HA3000) HA were labeled with fluorescein and injected into the knees of rabbits. Low-intensity continuous ultrasound at 1 MHz, 400 mW/cm2 was applied to the knees for 10 min treatment bid. Synovial fluid analysis revealed increased absorption and fluorescence microscopy showed deeper penetration of both HA1000 and HA3000, more so with the latter. Histologic examination indicated that ultrasound treatment resulted in no apparent damage to the synovial membrane. These results suggest that simultaneous sonication with HA injection might compensate for the short half-life of HA. Consequently, this dual treatment would render HA a far more effective tool in the management of OA.

(R)- and (S)-Epichlorohydrins were used to prepare the enantiomers of sterically diverse haloacetylcarbamoyl-2-nitroimidazoles that function as hypoxic cell radiosensitizers. The synthetic design allowed for introduction of a side chain of varying bulk that permitted an examination of the steric effects on enantio-discrimination in biological assay systems. The single stereocenter also connected the two pharmacophores––a 2-nitroimidazole moiety critical to hypoxic cell radiosensitization, and a haloacetylcarbamoyl group to function as an anti-angiogenesis pharmacophore. In the chick embryo chorioallantoic membrane (CAM) assay, the R-enantiomers possessing the bulky p-tert-butylphenyl group showed higher anti-angiogenic activity than the corresponding S-enantiomers, while there were no differences in the activity between the enantiomers containing the less bulky methyl and tert-butyl groups. Among the compounds we report, R-p-tert-butylphenyl-bromoacetylcarbamoyl-2-nitroimidazole, TX-1898, was found to be the most promising candidate for further development of as anti-angiogenic hypoxic cell radiosensitizer.

A lot of non-viral gene delivery carriers including cationic lipids and polymers have been developed due to their advantages such as easy introduction of specific bio-functionalities, convenient handling, non-immunogenicity and unlimited capacity of genes delivered. Poly(ethylenimine) (PEI) and poly(amido amine) (PAMAM) dendrimer are representative and commercialized polymeric gene delivery carriers. PEI shows very high transfection efficiency caused by its ‘endosome buffering’ capacity but its severe cytotoxicity limited its application for gene delivery and led its chemical modification for reducing the cytotoxicity. Recently, interesting strategies based on the preincubation of potent glucocorticoid, dexamethasone before transfection and the conjugation of mitochondrial leader peptide to PEI have been also tried in order to enhance the transfection efficiency of PEI by glucocorticoid ligand-receptor interaction and to perform selective delivery of genes to mitochondria, respectively. The transfection efficiency of PAMAM dendrimer is lower than that of PEI. However, its controllable multivalent functionality, defined structure and biocompatibility is still arising many scientists’ interest for gene delivery systems. So, we intended to design a novel hybrid dendrimer, poly(amido ethylenimine) (PAMEIM) dendrimer possessing the endosome buffering capacity of PEI and the defined structure, biocompatibility of PAMAM dendrimer in order to combine their advantages. Here, we report synthesis of a novel PAMEIM dendrimer and characterization of its self-assembly with plasmid DNA in order to identify its potential as a gene delivery carrier.

We designed, synthesized, and evaluated haloacetylcarbamoyl-2-nitroimidazoles, including chloro (KIN-1800, TX-1835, and TX-1836) and bromo derivatives (TX-1844, TX-1845, and TX-1846), as potential hypoxic cell radiosensitizers with antiangiogenic activities. To establish biological function owing to the haloacetylcarbamoyl group in the side-chain, we compared their in vitro radiosensitizing activities with those of their parent 2-nitroimidazoles without haloacetylcarbamoyl groups: misonidazole (MISO), TX-1831, and TX-1832, respectively. Both tert-butoxy substituted derivatives. TX-1835 and TX-1845, were more potent radiosensitizers than TX-1831. The p-tert-butylphenoxy-substituted derivatives, TX-1836 and TX-1846, and the methoxysubstituted derivatives, KIN-1800 and TX-1844, were stronger radiosensitizers than TX-1832 and MISO. We examined the anti-angiogenic activities of these 2-nitroimidazole derivatives containing haloacetylcarbamoyl group by the rat lung endothelial (RLE) cell proliferation assay and chick embryo chorioallantoic membrane (chick CAM) angiogenesis assay and showed that haloacetylcarbamoyl-2-nitroimidazoles were more potent angiogenic inhibitors than the corresponding desacetylcarbamoyl-2-nitroimidazoles. The in vivo chick CAM angiogenesis assay showed that the strong bromoacetylcarbamoyl-2-nitroimidazole radiosensitizers, such as TX-1845 and TX-1846, were the strongest angiogenic inhibitors among them. We concluded that the bromoacetylcarbamoyl-2-nitroimidazole radiosensitizers, such as TX-1845 and TX-1846, are promising as anti-angiogenic hypoxic cell radiosensitizers.

The changes in the gas permeability and permselectivity of natural zeolite disk by heat treatment were examined. The natural zeolite disks were fired for several hours at temperatures of 673 to 1273 K. The gas permeability and separation factor of the disks were measured by the gas permeation experiments with nitrogen-hydrogen gaseous mixture. The pore size distributions of the disks were evaluated by mercury porosimetry. The gas permeation experiments showed that gas permeability and separation factor simultaneously improved under a proper heat treatment condition. However, the measured mean pore diameters of the heat-treated samples was found to increase as the heat treatment temperature was increased. Such phenomenon could not be interpreted with considering conventional gas molecule transport mechanism through straight pores. Based on the observations, a qualitative model for pore structure change by neck growth due to sintering was proposed to account for the simultaneous improvement of the gas permeability and separation factor.

BACKGROUND: Calcium phosphate cement (CPC) can be molded or injected to form a scaffold in situ, which intimately conforms to complex bone defects. Bioactive glass (BG) is known for its unique ability to bond to living bone and promote bone growth. However, it was not until recently that literature was available regarding CPC-BG applied as an injectable graft. In this paper, we reported a novel injectable CPC-BG composite with improved properties caused by the incorporation of BG into CPC. MATERIALS AND METHODS: The novel injectable bioactive cement was evaluated to determine its composition, microstructure, setting time, injectability, compressive strength and behavior in a simulated body fluid (SBF). The in vitro cellular responses of osteoblasts and in vivo tissue responses after the implantation of CPC-BG in femoral condyle defects of rabbits were also investigated. RESULTS: CPC-BG possessed a retarded setting time and markedly better injectability and mechanical properties than CPC. Moreover, a new Ca-deficient apatite layer was deposited on the composite surface after immersing immersion in SBF for 7 days. CPC-BG samples showed significantly improved degradability and bioactivity compared to CPC in simulated body fluid (SBF). In addition, the degrees of cell attachment, proliferation and differentiation on CPC-BG were higher than those on CPC. Macroscopic evaluation, histological evaluation, and micro-computed tomography (micro-CT) analysis showed that CPC-BG enhanced the efficiency of new bone formation in comparison with CPC. CONCLUSIONS: A novel CPC-BG composite has been synthesized with improved properties exhibiting promising prospects for bone regeneration.

J Oral Pathol Med (2010) 39 182–187 Background: Candida albicans is the important opportunistic fungal pathogens which can cause oral Candidiasis and even more seriously systemic infection. Apoptosis of C. albicans induced by environmental factor such as weak acid and antifungal drugs were studied recently. Illustrating the phenomenon of apoptosis in C. albicans may help us to discover new antifungal therapy by activating the fungal cells to suicide. Methods: Two oral C. albians clinical isolates which isolated respectively from healthy host [Strain 23C: minimal inhibition concentration (MIC) is 0.125 μg/ml for Amphotericin B (AmB)] and advanced cancer patient (Strain 28A: MIC is 2 μg/ml for AmB), were induced by 1 μg/ml AmB in vitro for 200 min, and then studied the apoptosis markers using terminal deoxynucletidyltransferase-mediated dUTP nick end labeling (TUNEL) (shown by diaminobenzidine and fluorescent isothiocyanate), and the ultrastructure of cell nuclear using transmission electron microscope (TEM), quantitative analysis using flow cytometry for the rapid exposure of phosphatidylserine at the outer membrane and propodium iodide (PI) double staining. C. albicans conference strain YEM30 was used as the control strain. Results: With TUNEL assay and TEM, we detected the typical characteristics of apoptosis. Strain 23C (with low MIC) showed significantly higher percentage of apoptosis (19.92%) compared with Strain 28A (with high MIC) which was isolated from the cancer patient (7.29%) (P

The protein phosphatase 1 has conserved cores in PPP gene family flanked by non-conserved N-terminal domains. PP1 with residues 1-8 deleted or substituted by residues 1-42 of calcineurin catalytic subunit were designated PP1-(9-330) and CNA(1-42)-PP1(9-330), respectively. When compared with PP1, PP1-(9-330) had higher and CNA(1-42)-PP1(9-330) had lower activity with three kinds of substrates; PP1-(9-330) has higher and CNA(1-42)-PP1(9-330) has lower sensitivity to okadaic acid. These results imply that the N-terminal residues influence the activity and sensitivity to inhibitors of PP1. PP1-(9-330), PP1, and CNA(1-42)-PP1(9-330) displayed increasing K (m) and decreasing V (max) with three kinds of substrates, which suggest that the N-terminal residues are connected with the substrates affinity and catalytic efficiency of PP1. PP1-(9-330) has higher and CNA(1-42)-PP1(9-330) has lower fluorescence intensity than PP1, and the emission wavelength maximum was blue-shifted from PP1 to PP1-(9-330) and red-shifted from PP1 to CNA(1-42)-PP1(9-330). Our findings provide evidence that the N-terminal domain is an important region influencing the structure and properties of PP1.

The protein phosphatase 1 catalytic subunit (PP1c) and the protein phosphatase 2B (PP2B or calcineurin) catalytic subunit (CNA) contain nonconserved N-terminal regions followed by conserved phosphatase cores. To examine the role of the N-termini of these two phosphatases, we substituted the residues 1–8 of PP1c with residues 1–42 of CNA, which is designated CNA(1-42)-PP1(9-330). The activities of CNA(1-42)-PP1(9-330) were similar to those of PP2B and different from those of PP1. The chimera was at least fourfold less sensitive to inhibition by okadaic acid, but was stimulated by nickel ions and chlorogenic acid, characteristics of PP2B not of PP1. These observations suggest that the N-terminus of CNA shifts the properties of PP1 toward those of PP2B. Our findings provide evidence that the nonconserved N-terminus of PP2B not only functions as important regulatory domain but also confers itself particular characteristics. This region may be targeted for regulation of PP2B activities in vivo. © 2008 IUBMB IUBMB Life, 61(2): 178–183, 2009

Mono- and di-arginine conjugated PAMAM dendrimers (G = 3 or 4) were synthesized to examine the structure-activity relationships of arginine conjugation for gene delivery systems. Number of conjugated arginines was examined by 1H NMR (PAMAM3-R: 31, PAMAM4-R: 60, PAMAM3-R2: 59, PAMAM4-R2: 116). They could retard pDNA at a charge ratio of 2 and form polyplexes with sizes less than 250 nm from a charge ratio of 4. Di-arginine conjugated dendrimers showed higher Zeta-potential values and polyplex stabilities than mono-arginine conjugates. PAMAM3-R and PAMAM4-R showed low cytotoxicities even at high concentration but PAMAM3-R2 and PAMAM4-R2 exhibited significant cytotoxicities at high concentration. PAMAM3-R2 displayed greater transfection efficiency than PAMAM3-R, although the transfection efficiency of PAMAM4-R2 was not higher than that of PAMAM4-R in all condition. PAMAM3-R2 and PAMAM4-R2 polyplexes were observed to show the good intra-nuclear localization in comparison with PAMAM3-R and PAMAM4-R. It is concluded that di-arginine conjugation to PAMAM dendrimers can improve polyplex stability, intra-nuclear localization, and transfection efficiency but also induce charge density- and generation-dependent cytotoxicity. Therefore, a novel strategy for highly densed arginine conjugation maintaining low cytotoxicity will be needed for the development of efficient gene delivery carriers.

We investigated the synergistic effect of combined treatment with red ginseng acidic polysaccharide (RGAP) from Panax ginseng C.A. Meyer and pidotimod in cyclophosphamide-treated mice. The combination of pidotimod and RGAP restored concanavalin A-induced splenic T cell proliferation and LPS-stimulated B cell proliferation significantly. The production of nitric oxide from peritoneal macrophages was increased by the combinations. NK cell activity was increased by RGAP alone or in combination with pidotimod. A synergistic increase in the level of serum IL-12 and interferongamm was observed when the combination of the two was used. RGAP alone or in combination with pidotimod modulated the level of serum C-reactive protein to a near-normal level. These results indicate that combinations of pidotimod and RGAP are synergistic and suggest that combination therapy using pidotimod and RGAP for improving immune activity may provide an additional benefit over the use of the two drugs by themselves.

We have observed in our previous study that a cell-derived extracellular matrix (ECM) scaffold could assure the growth of a cartilage tissue construct in vitro. The purpose of the present study was to evaluate the feasibility of a chondrocyte-seeded cell-derived ECM scaffold by implanting it in vivo in nude mouse. A porous cell-derived ECM scaffold was prepared with a freeze-drying protocol using porcine chondrocytes. Rabbit articular chondrocytes were seeded onto the scaffold and cultured for 2 days in vitro, and then implanted into the nude mouse subcutaneously. They were retrieved at 1, 2, and 3 weeks postimplantation. Under macroscopic analysis, the cartilage-like tissue formation matured with time and developed a smooth, white surface. Contrary to the control (in which no cells were seeded), the size of the neocartilage tissue increased slightly by the third week and remained more stable. Total glycosaminoglycan (GAG) content and the GAG/DNA ratio increased significantly with time in the chemical analysis. The histology exhibited a sustained accumulation of newly synthesized sulfated proteoglycans. Immunohistochemistry, Western blot, and reverse transcriptase-polymerase chain reaction (RT-PCR) clearly identified type II collagen at all time points. Compressive strength of in vivo neocartilage increased from 0.45 +/- 0.06 MPa at 1 week to 1.18 +/- 0.17 MPa at 3 weeks. In conclusion, this study demonstrated that the cell-derived ECM scaffold could provide chondrocytes with favorable in vivo environment to produce a hyaline-like cartilage tissue.

2-Bromopropane (2-BP), a halogenated propane analogue, is a substitute for chlorofluorocarbones. The present study was carried out to investigate the potential adverse effects of 2-BP on pregnant dams and embryo-fetal development after maternal exposure on gestational days (GD) 6 through 19 in Sprague-Dawley rats. The test chemical was administered subcutaneously to pregnant rats at dose levels of 0, 250, 500, and 1000 mg/kg per day. All dams were subjected to caesarean section on GD 20 and their fetuses were examined for external, visceral and skeletal abnormalities. In the 1000 mg/kg group, maternal toxicity included an increase in the incidence of abnormal clinical signs, a suppression in the body weight and body weight gain, and a decrease in the food intake. Developmental toxicity included an increase in the fetal deaths, a decrease in the litter size, and a reduction in the fetal body weight. In addition, an increase in the incidence of fetal external, visceral, and skeletal abnormalities was seen. In the 500 mg/kg group, minimal developmental toxicity including decreased fetal body weight and increased fetal ossification delay was observed. There were no adverse effects on either pregnant dams or embryo-fetal development in the 250 mg/kg group. These findings suggest that a 14-day subcutaneous dose of 2-BP is embryotoxic and teratogenic at a maternally toxic dose (i.e., 1000 mg/kg per day) and is minimally embryotoxic at a nonmaternally toxic dose (i.e., 500 mg/kg per day) in Sprague-Dawley rats. In the present experimental conditions, the no-observed-adverse-effect level (NOAEL) of 2-BP is considered to be 500 mg/kg per day for dams and 250 mg/kg per day for embryo-fetal development.

New N-thiadiazolylanilines were designed and synthesized to develop mitochondrial cytotoxins superior to SF 6847. The mitochondrial cytotoxin N-thiadiazolylanilines, TX-108 and TX-109, inhibited EMT6/KU mammary sarcoma cell growth at a low micromolar concentration. Their inhibitory activities were parallel to their mitochondrial cytotoxicity, such as uncoupling oxidative phosphorylation and inhibiting ATP synthesis. This report also supports the notion that the inhibition of tumor cell growth of inhibitor of protein tyrosine kinase AG17, which is identical to SF 6847, may be due to its mitochondrial cytotoxicity.

Many more Na2 2 3pi(g) v = 0-43, omega = 0, 1, 2 levels have been observed by sub-Doppler continuous wave perturbation facilitated optical-optical double resonance fluorescence excitation spectroscopy and the hyperfine structure of the omega = 0 and 2 levels has been resolved. New molecular constants for the less perturbed v = 0-43 levels have been obtained with these new and the previously reported data. The hyperfine coupling scheme of the observed 2 3pi(g) levels is close to Hund's case a(beta) with a Fermi contact constant b(F) = 160+/-5 MHz, which is smaller than the Fermi contact constants of other Na2 triplet Rydberg states, b(F) = 200-245 MHz.