In vitro analysis of inflammatory responses following environmental exposure to pharmaceuticals and inland waters

Pharmaceuticals are regularly released into the environment; in particular non-steroidal anti-inflammatory drugs (NSAIDs) and antibiotics. Erythromycin, naproxen, furosemide and atenolol are reported to be stable for up to 1 year in the environment, which increases the risk for accumulation. In the present study we have measured the occurrence and concentration of pharmaceuticals in river Viskan (Jössabron) downstream of a sewage treatment plant in Borås, Sweden. Pharmaceuticals and water samples were tested for potential human risk by evaluating inflammatory responses (NF-κB and AP-1) using human T24 bladder epithelial cells and Jurkat T-cells. NF-κB activity in T24 cells was significantly reduced by all NSAIDs analysed (diclofenac, ketoprofen, naproxen, ibuprophen and dextropropoxyphene), but also by trimethoprim, using environmentally relevant concentrations. NF-κB and AP-1 activation was further analysed in response to water samples collected from different locations in Sweden. Dose-dependent down-regulation of AP-1 activity in Jurkat cells was observed at all locations. At two locations (Jössabron and Almenäs) down-regulation of NF-κB was observed. In contrast, the NF-κB response was potentiated by exposure to water from both locations following activation of NF-κB by treatment with heat-killed Escherichia coli. To determine the involvement of pharmaceuticals in the responses, T24 cells were exposed to the pharmaceutical mixture, based on the determined levels at Jössabron. This resulted in reduction of the NF-κB response following exposure to the pharmaceutical mixture alone while no potentiation was observed when cells were co-exposed to heat killed E. coli and pharmaceuticals. The obtained results demonstrate that the identified pharmaceuticals affect the inflammatory responses and furthermore indicate the presence of unknown substance(s) with the ability to potentiate inflammatory responses.     

硫酸镁对人脐静脉血管内皮细胞照射后NF-κB/ICAM-1的影响

为研究硫酸镁对粘附蛋白ICAM-1及其调控基因NF-κB表达的影响,进而探讨硫酸镁对人脐静脉血管内皮细胞(human umbilical vein endothelial,HUVEC)放射损伤的防护机制。取对数生长期细胞分为空白对照组、单纯照射组、照射加药组,单次4 Gy X射线照射单纯照射组及照射加药组,其中照射加药组于照射前0.5 h加入1.25 mg／mL硫酸镁,收集样本,利用流式细胞术和RT-PCR法检测NF-κB、ICAM-1 mRNA的表达情况,Western blot、激光共聚焦检测NF-κB、ICAM-1蛋白的表达情况。结果表明,照射后72 h内,NF-κB、ICAM-1 mRNA表达均高于空白对照组,其中照后3 h表达最为显著;在照射后48、72 h,1.25 mg/mL硫酸镁处理组中NF-κB / ICAM-1蛋白均低于单纯照射组。以上结果说明,电离辐射能够诱导NF-κB /ICAM-1高表达,而硫酸镁能抑制其表达,且可能是通过抑制NF-κB核转位来调控ICAM-1表达的途径参与辐射损伤防护。     

Epitaxial growth of Dy2O3 thin films on epitaxial Dy-germanide films on Ge(0 0 1) substrates

Ultra-thin films of Dy are grown on Ge(0 0 1) substrates by molecular beam deposition near room temperature and immediately annealed for solid phase epitaxy at higher temperatures, leading to the formation of DyGe_x films. Thin films of Dy₂O₃ are grown on the DyGe_x film on Ge(0 0 1) substrates by molecular beam epitaxy. Streaky reflection high energy electron diffraction (RHEED) patterns reveal that epitaxial DyGe_x films grow on Ge(0 0 1) substrates with flat surfaces. X-ray diffraction (XRD) spectrum suggests the growth of an orthorhombic phase of DyGe_x films with (0 0 1) orientations. After the growth of Dy₂O₃ films, there is a change in RHEED patterns to spotty features, revealing the growth of 3D crystalline islands. XRD spectrum shows the presence of a cubic phase with (1 0 0) and (1 1 1) orientations. Atomic force microscopy image shows that the surface morphology of Dy₂O₃ films is smooth with a root mean square roughness of 10 Å.     

Effect of pH adjustment on the composition and rennet-gelation properties of milk concentrates made from ultrafiltration and reverse osmosis

The objective of this work was to investigate the effect of pH adjustment (initial pH vs. pH 6.50) on the rennet-gelation properties of concentrates made by ultrafiltration (UF) and reverse osmosis (RO). Rennet-gelation kinetics were followed by dynamic rheology and κ-casein hydrolysis by reverse-phase HPLC. At initial pH, RO concentrates had better rennet-coagulation behavior than UF concentrates and skim milk, whereas adjusting the pH to 6.50 produced the opposite results. The kinetics of κ-casein hydrolysis were similar in skim milk, and both concentrates and were not affected by pH adjustment. Differences in rennet coagulation were then related to the extent of hydrolysis required to trigger casein micelle aggregation. Small pH adjustments (<0.2 pH unit) enabled the use of RO concentrate with similar rennet-gelation behavior to UF concentrate, despite major compositional differences. This study shows that pH adjustment of RO concentrates can be a simple approach to improve their coagulation properties; however, the mechanisms behind these improvements remain to be elucidated.     

蛋白酶体抑制剂MG132对肿瘤恶病质的作用及其分子机制

目的探讨蛋白酶体抑制剂MG132对肿瘤恶病质的作用及其可能的分子机制。方法经小鼠腋窝皮下注射结肠腺癌C26细胞,建立肿瘤恶病质模型,并设正常对照组(HC组)。将模型小鼠分为肿瘤恶病质组(CC组)和MG132治疗组(MG组),待小鼠进入恶病质状态后,CC组小鼠经腹腔注射0.1 ml生理盐水,MG组小鼠经腹腔注射0.1 mg/kg的MG132,7 d后处死小鼠,称量小鼠肿瘤、左侧腓肠肌和附睾脂肪的重量,测量腓肠肌纤维横切面积,ELISA法检测血清中炎性因子TNF-α和IL-6的水平,RT-PCR及Western blot法检测腓肠肌中IKBa、P65、MuRF1和MAFbx基因mRNA的转录水平及蛋白的表达水平。结果与CC组相比,MG组小鼠腓肠肌和附睾脂肪组织的重量分别增加了31.6%和39.5%(P<0.05),腓肠肌纤维横切面积增加了36.1%(P<0.05);血清中TNF-α和IL-6的水平分别降低了20.9%和42.0%(P<0.05);腓肠肌组织IKBa基因mRNA的转录水平和蛋白表达水平分别升高了132.7%和56.5%(P<0.05),MuRF1基因mRNA的转录水平和蛋白表达水平分别降低了70.1%和42.6%(P<0.05),MAFbx基因mRNA的转录水平和蛋白表达水平分别降低了76.8%和47.3%(P<0.05),P65基因mRNA的转录水平和蛋白表达水平分别降低了59.1%和53.1%(P<0.05)。结论 MG132改善肿瘤恶病质的分子机制可能与抑制NF-κB途径及MuRF1和MAFbx的表达、抑制炎症反应及肿瘤生长有关。     

Toona sinensis and its major bioactive compound gallic acid inhibit LPS-induced inflammation in nuclear factor-κB transgenic mice as evaluated by in vivo bioluminescence imaging

In the present study, we investigated the anti-inflammatory effects of a nutritious vegetable Toona sinensis (leaf extracts, TS) and its major bioactive compound gallic acid (GA) by analysing LPS-induced NF-κB activation in transgenic mice, using bioluminescence imaging. Mice were challenged intraperitoneally with LPS (1 mg/kg) and treated orally with TS or GA (100 or 5 mg/kg, respectively). In vivo and ex vivo imaging showed that LPS increased NF-κB luminescence in the abdominal region, which was significantly inhibited by TS or GA. Immunohistochemical and ELISA analyses confirmed that TS and GA inhibited LPS-induced NF-κB, interleukin-1β, and tumour necrosis factor-α expression. Microarray analysis revealed that biological pathways associated with metabolism and the immune responses were affected by TS or GA. Particularly, LPS-induced thioredoxin-like 4B (TXNL4B) 2 expression in the small intestine, and TXNL4B, iNOS, and COX-2 expression in RAW 264.7 cells were significantly inhibited by TS or GA. Thus, the anti-inflammatory potential of TS was mediated by the downregulation of NF-κB pathway.     

Magnesium metallic interlayer as an oxygen-diffusion-barrier between high-κ dielectric thin films and silicon substrate

The deposition of a thin magnesium metallic interlayer on an Si substrate prior to the deposition of an oxide thin film using rf-sputtering was investigated. The deposition of high-κ HfO₂ thin film was more particularly studied and it was demonstrated that the metallic interlayer acts as an oxygen barrier, preventing the formation of a low-κ layer at the high-κ/Si interface during the deposition. A post-deposition annealing treatment performed on the films induced the diffusion of the metal barrier into the HfO₂ film and allowed obtaining a sharp interface. However, the degree of diffusion depends not only on the interlayer thickness, but also on the thickness of the high-κ film. X-ray photoelectron spectroscopy was used to study the degree of oxidation of the Mg interlayer. High resolution transmission electron microscopy and energy filtered transmission electron microscopy were used to characterize the films and the diffusion of the Mg interlayer into the high-κ film after annealing. In this work we will stress on the engineering of the interface via the diffusion of the Mg interlayer during the growth process and on annealing.     

Detection of open or closed porosity in low-κ dielectrics by solvent diffusion

The presence of open porosity in a porous interlayer dielectric (ILD) can seriously degrade the ultimate performance and reliability of a device. A simple method for detecting the open porosity in ILDs would facilitate the design of materials with pore morphologies compatible with the required specifications of a particular device. The measurement of effective diffusion coefficients in a low-κ ILD over a range of porosities is shown to permit the detection of open porosity. An example illustrating the simplicity of this method is presented using toluene solvent in a well-characterized porous organosilicate material. The result using this method is consistent with available data in the literature.     

Super-connected but not super edge-connected graphs

A connected graph G is super-connected (resp. super edge-connected) if every minimum vertex-cut (resp. edge-cut) isolates a vertex of G. In [Super connectivity of line graphs, Inform. Process. Lett. 94 (2005) 191-195], Xu et al. shows that a super-connected graph with minimum degree at least 4 is also super edge-connected. In this paper, a characterization of all super-connected but not super edge-connected graphs is given. It follows from this result that there is a unique super-connected but not super edge-connected graph with minimum degree 3, that is, the Ladder graph L₃ of order 6, and that there are infinitely many super-connected but not super edge-connected graphs with minimum degree 1 or 2.