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In the present study, acute onset of severe lupus nephritis was successfully treated in mice using a new, benzamide-linked, small molecule that targets immune modulation and the NLRP3 inflammasome. Specifically, 6-(2,4-difluorophenyl)-3-(3-(trifluoromethyl)phenyl)-2H-benzo[e][1,3]oxazine-2,4(3H)-dione (Cf-02) (a) reduced serum levels of IgG anti-dsDNA, IL-1β, IL-6, and TNF-α, (b) inhibited activation of dendritic cells and differentially regulated T cell functions, and (c) suppressed the NF-κB/NLRP3 inflammasome axis, targeting priming and activating signals of the inflammasome. Moreover, treatment with Cf-02 significantly inhibited secretion of IL-1β in lipopolysaccharide-stimulated macrophages, but this effect was abolished by autophagy induction. These results recommend Cf-02 as a promising drug candidate for the serious renal conditions associated with systemic lupus erythematosus. Future investigations should examine whether Cf-02 may also be therapeutic in other types of chronic kidney disease involving NLRP3 inflammasome-driven signaling.

In this study, two kojic acid-methimazole (2-mercapto-1-methylimidazole, MMI, 1) derivatives, 5-hydroxy-2-{[(1-methyl-1H-imidazol-2-yl)thio]methyl}-4H-pyran-4-one (compound 4) and 5-methoxy-2-{[(1-methyl-1H-imidazol-2-yl)thio]methyl}-4H-pyran-4-one (compound 5), were synthesized to examine their inhibitory kinetics on mushroom tyrosinase. Compound 4 exhibited a potent inhibitory effect on monophenolase activity in a dose-dependent manner, with an IC50 value of 0.03 mM. On diphenolase activity, compound 4 exhibited a less inhibitory effect (IC50 = 1.29 mM) but was stronger than kojic acid (IC50 = 1.80 mM). Kinetic analysis indicated that compound 4 was both as a noncompetitive monophenolase and diphenolase inhibitor. By contrast, compound 5 exhibited no inhibitory effects on mushroom tyrosinase activity. The IC50 value of compound 4 for the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity was 4.09 mM, being much higher than the IC50 of compound 4 for inhibiting the tyrosinase activity. The results indicated that the antioxidant activity of compound 4 may be partly related to the potent inhibitory effect on mushroom tyrosinase. Compound 4 also exerted a potent inhibitory effect on intracellular melanin formation in B16/F10 murine melanoma cells, and caused no cytotoxicity. Furthermore, compound 4 induced no adverse effects on the Hen's egg test-chorioallantoic membrane (HET-CAM).

Background Various physical and chemical methods for synthesis of metal nanoparticles have had some drawbacks. Therefore, green synthesis of gold nanoparticles (Au NPs) has became one of the most crucial emerging areas of nanobiotechnology. In the present study, plant-mediated synthesis of Au NPs was performed using Djulis (Chenopodium formosanum) shell extract as a reducing and stabilizing agent. Results Reaction parameters were manipulated to optimize the Au NPs using a UV-visible spectrophotometer. Optimized Au NPs with a surface plasmon resonance band at 533 nm were prepared using a 744 µg mL-1 extract and a solution of pH 2.62 chloroauric acid (HAuCl4 ·3H2 O) at 40 °C. High-resolution transmission electron microscopy (HR-TEM) results indicated that most of the resultant Au NPs were spherical in shape and exhibited a mean size of 8 ± 6 nm. Energy-dispersive X-ray spectroscopy (EDS), and selected area electron diffraction (SAED), and X-ray diffraction (XRD) confirmed the elemental gold and crystalline nature of the resultant NPs. FTIR spectrum analysis indicated the critical role of phenolic groups in the reduction of Au3+ ions and stabilization of the formed Au NPs. Moreover, the synthesized Au NPs possessed antibacterial activity for Escherichia coli and Staphylococcus aureus. Conclusion In this study, Au NPs were synthesized with high efficiency and stability using Djulis shell extract, and related antibacterial applications were demonstrated. © 2019 Society of Chemical Industry.

Triptolide, a traditional Chinese medicine, obtained from Tripterygium wilfordii Hook F, has anti-inflammatory, antiproliferative, and proapoptotic properties. We investigated the potential efficacy of triptolide on murine leukemia by measuring the triptolide-induced cytotoxicity in murine leukemia WEHI-3 cells in vitro. Results indicated that triptolide induced cell morphological changes and induced cytotoxic effects through G0/G1 phase arrest, induction of apoptosis. Flow cytometric assays showed that triptolide increased the production of reactive oxygen species, Ca2+ release and mitochondrial membrane potential (ΔΨm ), and activations of caspase-8, -9, and -3. Triptolide increased protein levels of Fas, Fas-L, Bax, cytochrome c, caspase-9, Endo G, Apaf-1, PARP, caspase-3 but reduced levels of AIF, ATF6α, ATF6β, and GRP78 in WEHI-3 cells. Triptolide stimulated autophagy based on an increase in acidic vacuoles, monodansylcadaverine staining for LC-3 expression and increased protein levels of ATG 5, ATG 7, and ATG 12. The in vitro data suggest that the cytotoxic effects of triptolide may involve cross-talk between cross-interaction of apoptosis and autophagy. Normal BALB/c mice were i.p. injected with WEHI-3 cells to generate leukemia and were oral treatment with triptolide at 0, 0.02, and 0.2 mg/kg for 3 weeks then animals were weighted and blood, liver, spleen samples were collected. Results indicated that triptolide did not significantly affect the weights of animal body, spleen and liver of leukemia mice, however, triptolide significant increased the cell populations of T cells (CD3), B cells (CD19), monocytes (CD11b), and macrophage (Mac-3). Furthermore, triptolide increased the phagocytosis of macrophage from peripheral blood mononuclear cells (PBMC) but not effects from peritoneum. Triptolide promoted T and B cell proliferation at 0.02 and 0.2 mg/kg treatment when cells were pretreated with Con A and LPS stimulation, respectively; however, triptolide did not significant affect NK cell activities in vivo. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 550-568, 2017.

This study investigated the polyphenol content, antioxidant activity, and inhibition ability of mushroom tyrosinase and melanogenesis of Dendrobium tosaense (DT) extract. Ground DT was extracted using deionized water (W) or 50% ethanol (50E) at room temperature (RT) or 50 &deg, C (50T) for 20 min. The 50T + 50E extract exhibited the highest total phenol content 47.0 &plusmn, 4.0 mg gallic acid equivalent/g DT extract, the highest level of 2,2&prime, azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) free-radical scavenging 66.0 &plusmn, 3.0 mg Trolox equivalent/g DT extract, and the highest reducing power 12.00 &plusmn, 0.50 mg vitamin C equivalent/g DT extract. The RT + W extract had the highest total flavonoid content 110.0 &plusmn, 3.0 mg quercetin equivalent/g DT extract. The RT + 50E extract had the lowest half maximal inhibitory concentration 1.30 &plusmn, 0.00 mg/mL for 2,2-diphenyl-1-picrylhydrazyl free-radical scavenging, and the lowest half maximal inhibitory concentration 6.40 &plusmn, 0.30 mg/mL for mushroom tyrosinase inhibition activity. DT extracts, especially RT + W and 50T + W, exhibited potent inhibitory effects on melanogenesis of B16/F10 cells. These results demonstrated the application potential of DT extract for skincare.

Summary Traditionally, tea is steeped in boiled water in Taiwan. In this study, Bi Luo Chun (250 μg mL−1) steeped at 4, 22, 30, 40 and 50 °C reached the saturation of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging capacity at the time of 15, 8, 2, 0.17 (10 s) and 0.17 min (10 s), respectively. The result indicated that tea steeped in cold water (under 22 °C) still can reach a saturation of antioxidant capacity in 15 min. However, the initial scavenging rate (% min−1) was quick and decreased with time at all steeping temperatures (r: −0.98 to −1.00). Total phenols and the DPPH scavenging capacity of Bi Luo Chun showed excellent correlation with time. However, the plot of DPPH scavenging capacity vs. total phenols was not always correlated. The result indicated that polyphenol infusion at different times may have a different DPPH scavenging capacity.

The acidic-ethanol anthocyanin extraction process of TNG73 sweet potatoes was improved with introducing the sonication treatment. The linear terms of sonication time and extraction temperature were determined to be significant factors. Response surface methodology (RSM) models were successfully established to perform the extraction study and energy requirement analysis. After calculating the required energy for collected anthocyanins (J/µg), the process with higher extraction temperature became less effective. Based on the combined results of extraction study and energy requirement analysis, the recommended operating condition of anthocyanin extraction process was at 25°C with 22 min sonication.

The biological effects of specific wavelengths, so-called “far-infrared radiation” produced from ceramic material (cFIR), on whole organisms are not yet well understood. In this study, we investigated the biological effects of cFIR on murine melanoma cells (B16-F10) at body temperature. cFIR irradiation treatment for 48 h resulted in an 11.8% decrease in the proliferation of melanoma cells relative to the control. Meanwhile, incubation of cells with cFIR for 48 h significantly resulted in 56.9% and 15.7% decreases in the intracellular heat shock protein (HSP)70 and intracellular nitric oxide (iNO) contents, respectively. Furthermore, cFIR treatment induced 6.4% and 12.3% increases in intracellular reactive oxygen species stained by 5-(and 6)-carboxyl-2′,7′-dichlorodihydrofluorescein diacetate and dihydrorhodamine 123, respectively. Since malignant melanomas are known to have high HSP70 expression and iNO activity, the suppressive effects of cFIR on HSP70 and NO may warrant future interest in antitumor applications.

This study reports the preparation of four varieties of water extract from sweet potato leaves from Taiwan, including TNG10, TNG57, TNG66 and YSP, and evaluates their antioxidative activity. The EC50 values (scavenging DPPH radicals) of TNG10, TNG57, TNG66 and YSP were 0.27 ± 0.01, 0.19 ± 0.01, 0.41 ± 0.02, and 0.31 ± 0.02 mg/ml, respectively, on a freeze-dry weight basis. The total phenolic contents of these water extracts were in the order: TNG57 > TNG10 > TNG66 > YSP. The TNG10 and TNG57 extracts exhibited better reducing power and scavenging effects of superoxide radicals than did TNG66 and YSP. At a concentration of 1 mg/ml, TNG10 and TNG57 significantly protected HaCaT cells from H2O2-induced cytotoxicity. The water extracts of YSP had more flavonoids than had those of TNG66 which may have contributed to their higher activity in many antioxidative assays. These results suggest that the water extracts of all four varieties of sweet potato leaves, and especially TNG10 and TNG57, display potent antioxidative effects.

Anthocyanins were successfully extracted from TNG73 purple sweet potatoes (PSP), which were provided by Agricultural Research Institute (ARI), Chia-Yi Agricultural Experiment Station, Taiwan. An acidic-ethanol extraction method was used, and the extraction parameters were carefully evaluated. The experimental design and process data for anthocyanin extraction were analyzed by statistical software. The HCl concentration, extraction time, and extraction temperature were significant factors in the extraction of anthocyanins from PSP. Using 15% HCl (1.5N HCl in ethanol) with a 9 g/100mL PSP solid/solvent ratio at 25°C for 30 min was adequate for PSP anthocyanin extraction. The extraction performance was increased 15~25% by running a second extraction. Under the same process conditions, the performance was improved about 80% when sonication was applied. The difference caused by the sonication treatment was more significant for the high solid/solvent ratio system. Ultrasound-assisted extraction was a useful tool for PSP anthocyanin extraction. The goals of this study were to collect detailed process data for further process scaling-up and to develop an ultrasound-assisted anthocyanin extraction process.

The chemical composition and functional activities of cold-pressed and water distilled peel essential oils ofCitrus paradisi(C. paradisi) andCitrus grandis(L.) Osbeck (C. grandis) were investigated in present study. Yields of cold-pressed oils were much higher than those of distilled oils. Limonene was the primary ingredient of essential oils ofC. paradisi(cold 92.83%; distilled 96.06%) andC. grandis(cold 32.63%; distilled 55.74%). In addition,C. grandisoils obtained were rich in oxygenated or nitrogenated compounds which may be involved in reducing cardiovascular diseases or enhancing sleep effectiveness. The order of free radical scavenging activities of 4 citrus oils was distilledC. paradisioil > cold-pressedC. paradisioil > distilledC. grandisoil > cold-pressedC. grandisoil. Cold-pressedC. grandisoil exhibited the lowest activity in all antioxidative assays. The order of antimicrobial activities of 4 citrus oils was distilledC. grandisoil, cold-pressedC. paradisioil > distilledC. paradisioil > cold-pressedC. paradisioil. Surprisingly, distilledC. grandisoil exhibited better antimicrobial activities than distilledC. paradisioil, especially againstEscherichia coliandSalmonella entericasubsp. The results also indicated that the antimicrobial activities of essential oils may not relate to their antioxidative activities.

A comparison is made of predictions of the structural evolution of Al 2 O 3 SiO 2 SiCC refractories during firing and under slag attack made with the FACT computation package, and experimental observations including the development of a decarburised zone, relative wear of different materials and phases present as determined by microstructural and X-ray examination. The corrosion tests were carried out under oxidising conditions and with slags both containing and free of iron oxide. Two different base refractory compositions were used, one containing clay and one without. The additives considered included 5% Al, 10% Si, 5% B 4 C, 5% BN, 5%B 2 O 3 , and 15% Si 3 N 4 . Experimentally, good wear resistance was associated with the predicted content of SiC in the refractory in the as-fired condition. The most effective additives were silicon and B 4 C, whilst the most deleterious was B 2 O 3 . These results are interpreted in terms of the thermochemical predictions, including the variation in oxygen potential with slag attack and it is shown that the materials can be ranked in the same order as observed in the wear tests.

We investigated the kinetics of 4N-acetyl-pentapeptides, Ac-P1, Ac-P2, Ac-P3, and Ac-P4, regarding inhibition of mushroom tyrosinase activity. The peptides sequences of Ac-P1, Ac-P2, Ac-P3, and Ac-P4 were Ac-RSRFK, Ac-KSRFR, Ac-KSSFR, and Ac-RSRFS, respectively. The 4N-acetyl-pentapeptides were able to reduce the oxidation ofL-DOPA by tyrosinase in a dose-dependent manner. Of the 4N-acetyl-pentapeptides, only Ac-P4 exhibited lag time (80 s) at a concentration of 0.5 mg/mL. The tyrosinase inhibitory effects of Ac-P4 (IC500.29 mg/mL) were more effective than those of Ac-P1, Ac-P2, and Ac-P3, in which IC50s were 0.75 mg/mL, 0.78 mg/mL, and 0.81 mg/mL, respectively. Kinetic analysis demonstrated that all 4N-acetyl-pentapeptides were mixed-type tyrosinase inhibitors. Furthermore, 0.1 mg/mL of Ac-P4 exhibited significant melanogenesis inhibition on B16F10 melanoma cells and was more effective than kojic acid. The melanogenesis inhibition of Ac-P4 was dose-dependent and did not induce any cytotoxicity on B16F10 melanoma cells.

Oxidative stress, including Ultraviolet (UV) irradiation-induced skin damage, is involved in numerous diseases. This study demonstrates that water extract ofEclipta prostrataL. (WEP) has a potent effect in scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH), superoxide radicals, and chelating ferrous ion, exhibiting IC50values of 0.23 mg/mL, 0.48 mg/mL, and 1.25 mg/mL, respectively. The WEP total phenol content was 176.45 mg gallic acid equivalents (GAE)/g sample. Chlorogenic acid, a component of the plant's active ingredients, was determined by HPLC and antioxidative assay. However, no caffeic acid, stigmasterol, or wedelolactone was present in WEP. WEP absorbs both UVA and UVB irradiation, and furthermore, the extract shows a dose-dependent response in the protection of HaCaT human keratinocytes and mouse fibroblasts 3T3 cells against UVB-induced cytotoxicity, which may result from a synergistic effect between chlorogenic acid and other active components present in WEP.

Dequalinium (DQ, an anticancer drug) is a potent neurotoxicant in the cultured developing cerebellar granule neurons (CGNs) with an IC(50) of 1.31. M after 24 hr incubation. By utilizing fluorometric technique, we found that DQ initially induced apoptosis and then necrosis associated with a marked decrease in ATP contents. The purinergic P(2) receptor antagonists (suramin, and reactive blue 2) prevented DQ-cytotoxicity, although glutamate ionotropic receptor antagonists (MK 801 and NBQX) could not. Furthermore, we quantitatively determined a reduction of mitochondrial membrane potential and an increase of free radical production induced by DQ. Suramin abolished these detrimental events of DQ. This suggests that neuronal death induced by DQ is mediated, at least in part, through a signaling pathway of free radical production-mitochondrial dysfunction. Further evidence supporting this contention is that CGN progressively became more sensitive to both DQ-induced cytotoxicity and reduced mitochondrial membrane potential. This implies that neuronal mitochondria are apparently one of the target sites for DQ and suramin and directly or indirectly induce neurotoxicity and neuroprotection respectively. The alteration in mitochondrial membrane potential during neuronal maturation may be one of the determinants accounting for the increased susceptibility to neurotoxicants such as DQ.

An InAs-GaAs quantum ring infrared photodetector has been fabricated successfully. The photodetector demonstrates a cutoff wavelength at 175 mum (1.7 THz) and the detectivity of 1.3times107 cm Hz1/2/W at 80 K. The precise control of the In(Ga)As ring height by capping GaAs layer thickness is responsible for extension of the detector response to terahertz range.

Three sweet potato cultivars (TNG57, TNG66, and TNG73), provided by the Taiwanese Agricultural Research Institute (TARI), were stored at either 15 °C or under ambient conditions (23.8 ~ 28.4 °C and 77.1 ~ 81.0 % of relative humidity). Sweet potato roots were randomly chosen from each replicate and evaluated for measurement of weight loss, sugar content analysis, and sprouting after 0, 14, 24, 48, 56, 70, 84, and 98 days of storage. Fresh sweet potato roots were baked at 200 °C for 60 min then samples were taken for sugar analysis. After 14 days of ambient condition storage, the sprouting percentages for TNG57, TNG66, and TNG73 were 100, 85, and 95 % respectively. When sweet potatoes were stored at 15 °C, the weight loss became less and no sweet potato root sprouted after 14 days of storage. Because manufacturers can store sweet potatoes at 15 °C for almost 2 month without other treatments, the supply capacity shortage in July and September can be reduced. The total sugar content slowly increased along with increasing the storage time. After baking, the total sugar content of sweet potatoes significantly increased due to the formation of maltose. Maltose became the major sugar of baked sweet potatoes. Raw sweet potatoes stored at 15 °C had higher total sugar contents after baking than those stored under ambient conditions. Raw sweet potatoes were recommended to be stored at 15 °C before baking.

The objective of this study was to evaluate the influence of sugar composition on the sensory attributes of seven baked sweet potatoes. The sugar composition was analyzed using high performance liquid chromatography. Results showed that the total sugar content of baked sweet potatoes increased significantly because of the formation of maltose. The maltose content dramatically increased after baking, and became the major sugar component of baked sweet potatoes. On the other hand, baked sweet potatoes were evaluated on a 7-point hedonic scale for sensory analysis. Overall acceptability results showed that the panelists preferred baked CYY95-26 and TNG66 over the other baked varieties. Because the correlation between overall acceptability and sweetness was the highest (r = 0.69, p

We investigated the relationship between [Ca2+]i, myosin light chain (LC20) phosphorylation and isometric force in guinea pig aortic strips during contractions activated by a thromboxane A2 analogue, (15S)-hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5Z, 13E-dienoic acid (U46619). Isometric force and [Ca2+]i were measured simultaneously using preloaded aequorin as the intracellular calcium indicator. LC20 phosphorylation levels were determined by two dimensional polyacrylamide gel electrophoresis in parallel preparations. Contractions induced by U46619 were accompanied by increases in [Ca2+]i and LC20 phosphorylation. The chelation of extracellular calcium with 2.5 mM EGTA significantly inhibited U46619-induced increases in [Ca2+]i, isometric force and LC20 phosphorylation. Steady-state force assumed a similar dependence on LC20 phosphorylation for contractions stimulated by potassium depolarization, alpha 1-adrenergic agonist phenylephrine and U46619 either in the presence or absence of extracellular calcium. On the contrary, the [Ca2+]i/force relation revealed that both U46619 and phenylephrine stimulated greater isometric force at lower [Ca2+]i than did KCl depolarization. The addition of a protein kinase C inhibitor, 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7), decreases force without significantly affecting either [Ca2+]i or LC20 phosphorylation levels. These results suggest that in guinea pig aortic smooth muscle U46619 increases the calcium sensitivity of the contractile apparatus but does not change the LC20 phosphorylation/force relation in comparison to K+ depolarization. Protein kinase C is activated during U46619-stimulated contraction and might be involved in mechanisms other than LC20 phosphorylation.

Ceramic bodies of mullite composition were prepared from a mixture of calcined alumina and raw kaolin with the addition of bastnasite (rare-earth oxide concentrate) at up to 3% wt and fired at 1400–1650°C. The bastnasite addition effectively enhances mullite formation but the effect diminishes rapidly with increasing temperature. The Al 2 O 3 -rich composition is characteristic of the mullite. The ceramic green compacts with the bastnasite addition can only approx. reach 96% theoretical density at best when heated at 1550–1650°C. The pores in the fired bodies with the bastnasite addition are smaller and uniformly distributed; localized, larger pores are found in the fired bodies without the addition. The slag resistance of the refractories increases slightly with an increasing amount of bastnasite added to the bonding matrix. Bastnasite is not an effective additive for enhancing the slag resistance of steel-plant refractories. The peak temperature in DTA and the valley temperature in the dilatometric analysis, are indicative of mullite formation: the former was found to be 1412°C without bastnasite addition and 1368°C with 3% wt bastnasite addition while the latter was found to be 1420°C without bastnasite addition and 1350°C with 3% wt bastnasite addition.

Sugar composition of seven sweet potato cultivars was successfully analyzed. Fresh CYY95-26 sweet potatoes had the highest (8.41%) total sugar content while TNG73 had the lowest (4.5%). For these fresh sweet potatoes, maltose content was very low (0 ~ 0.39%). Because 49.92 ~ 92.43% of total sugars were sucrose, sucrose was the major sugar composition of fresh sweet potatoes. After the baking treatment, the total sugar content of baked sweet potatoes was dramatically increased due to the formation of maltose. The maltose content significantly increased from 0 ~ 0.39% to 8.81 ~ 13.97% on dry weight basis. Therefore, maltose should be included in calculating the total sugar content. Electronic micrographs of fresh sweet potato samples showed that the size of starch granules was generally less than 20 μm. After the baking treatment, starch granules completely gelatinized.

The native amino acid ergothioneine, a thiourea derivative of histidine, inhibits mushroom tyrosinase activity in a dose-dependent manner, with an IC(50) value of 1.025 mg/ml (4.47 mM). By contrast, histidine exhibited no inhibitory effect on mushroom tyrosinase activity. We characterized ergothioneine as a noncompetitive tyrosinase inhibitor using a Lineweaver-Burk plot of experimental kinetic data. The IC(50) value for ergothioneine scavenging of 2,2-diphenyl-1-picrylhydrazyl was 6.110 ± 0.305 mg/ml, much higher than the IC(50) for inhibition of tyrosinase activity which indicating ergothioneine on tyrosinase shows a weak correlation to its antioxidative activity. The results demonstrated that ergothioneine has a potent inhibition effect on tyrosinase enzyme activity, resulting from the presence of the sulfur substituted imidazole ring in ergothioneine.

AIM: The genomes of Helicobacter pylori (H. pylori) from different individuals are different. This project was to identify the strain specific DNA sequences between two clinical H. pylori isolates by suppression subtractive hybridization (SSH). METHODS: Two clinical H. pylori isolates, one from gastric ulcer (GU, tester) and the other from non-ulcer dyspepsia (NUD, driver), were cultured and the genomic DNA was prepared and submitted to Alu I digestion. Then two different adaptors were ligated respectively to the 5’-end of two aliquots of the tester DNA fragments and SSH was made between the tester and driver DNA. The un-hybridized tester DNA sequences were amplified by two sequential PCR and cloned into pGEM-T-Easy Vector. The tester strain specific inserts were screened and disease related DNA sequences were identified by dot blotting. RESULTS: Among the 240 colonies randomly chosen, 50 contained the tester strain specific DNA sequences. Twenty three inserts were sequenced and the sizes ranged from 261 bp to 1036 bp. Fifteen inserts belonged to the H.pylori plasmid pHPO100 that is about 3.5 kb and codes a replication protein A. Other inserts had patches of homologous to the genes of H.pylori in GenBank. Various patterns of dot blots were given and no GU strain unique DNA sequences were found when 4 inserts were used as probes to screen the genomic DNA from 27 clinical isolates, 8 from GU, 12 from duodenum ulcer (DU), 4 from GU-DU, 2 from NUD and 1 from gastric cancer (GC). But a 670 bp DNA fragment (GU198) that was a bit homologous to the 3’-end of the gene of thymidylate kinase was positive in 7 GU strains (7/8), 3 GU-DU strains (3/4) and 3 DU strains (3/12). A 384 bp fragment (GU79) of the replication gene A (repA) was positive only in 4 H.pylori isolates, 2 from GU and 2 from GU-DU. CONCLUSION: Differences exist in the genes of different H.pylori isolates. SSH is very effective to screen H.pylori strain specific DNA sequences between two clinical isolates, and some of these sequences may have clinical significance.

In this study, we demonstrated that an anticancer drug, dequalinium, a bisquaternary ammonium compound, is a potent neurotoxicant with IC 50 of 0.46 μM on the cultured cerebellar granule neurons. Its selective neurotoxicity revealed by 100-fold more toxic than the other two analogs, pancuronium and vecuronium. The mechanisms underlying dequalinium (DQ)-induced neurotoxicity were explored and found to be associated with decreased mitochondrial membrane potential, increased free radical production and ATP depletion. Suramin (a nonselective purinergic P 2 receptor antagonist and an anticancer drug) but not the glutamate receptor antagonists, MK-801, NBQX (1,2,3,4 tetrahydro-6-nitro-2,3-dioxo-benzo[f]quinoxaline-7-sulfonamide disodium), and DNQX (6,7-dinitroquinoxaline-2,3-dione) significantly prevents the DQ-induced neurotoxicity. By means of microfluorometric image-processing technique using the fluorescent probes, fluorescein diacetate/propidium iodide and Hoechst 33258, respectively, we showed that 1 μM DQ for 24 h induced about 53.5% of apoptosis and 37.5% of necrosis. All of these effects of DQ can be completely prevented by suramin. From these results, we conclude that DQ-induced neurotoxicity was not mediated by glutamate receptor, but by increasing free radical productions and cell energy depletion. Suramin with its beneficial antagonistic effects on DQ-induced neurotoxicity may provide an effective approach for neurodegeneration.

In the title compound, C(13)H(21)N(3)O(4)S, the mean plane of the -N(H)-C(=O)-O- group of the carbamate unit forms a dihedral angle of 64.7 (2)° with the mean plane of the -C-C(=O)-O- group of the ester unit. In the crystal, mol-ecules are linked by N-H⋯O=C hydrogen bonds, forming chains along the c-axis direction.