Japanese encephalitis virus envelope protein mitigates TNF-alpha mRNA expression in RAW264.7 cells

[[abstract]]Japanese encephalitis virus (JEV) is known as an important mosquito-borne human pathogen that causes Japanese encephalitis and may lead to lethal effect. Since monocyte has been demonstrated to play transmissible role for JEV, rare study is reported to clarify the effect of JEV envelope (JEVE) protein on monocyte. This study intends to investigate the effects of JEVE protein inside monocyte. Notably, significant decreased expression of tumour necrosis factor (TNF)-alpha mRNA in RAW264.7 cells transfected with pEGFP-JEVE was observed as compared to those cells transfected with pEGFP. Increased p21(Waf1/Cip1) protein was observed in both pEGFP and pEGFP-JEVE transfected RAW264.7 cells. However, increased p53 protein was only detected in pEGFP-transfected cells but not pEGFP-JEVE transfected cells as well as the result that no increased expression of nuclear factor-kB was observed in pEGFP-JEVE transfected cells. These experimental results indicate the effects of JEVE protein in alleviating TNF-alpha mRNA expression that is associated with the increased p53-independent p21(Waf1/Cip1) expression and provide an explanation in the role of JEV transmission through monocyte

Emodin Has Cytotoxic and Protective Effects in Rat C6 Glioma Cells: Roles of Mdr1a and Nuclear Factor kappa B in Cell Survival

[[abstract]]1,3,8-Trihydroxy-6-methylanthaquinone (emodin) is recognized as an antiproliferative compound. In the present study, however, we show that emodin has both toxic and survival effects in glioma cells and that the survival effects involve Mdr1a. Emodin inhibited the proliferation and induced apoptosis of C6 cells in a 12-h treatment, but C6 cells survived a 72-h drug treatment, indicating resistance to emodin. Emodin-induced apoptosis was reduced by inhibition of the expression and activation of apoptosis-associated proteins including p53, Bax, Bcl-2, Fas, and caspase-3. C6 cells could express antioxidant proteins (superoxide dismutase and catalase) to decrease reactive oxygen species-induced cytotoxicity of emodin and overexpress multidrug resistance genes (Mdr1a, MRP2, MRP3, and MRP6) to decrease the intracellular accumulation of emodin. Electrophoretic mobility shift analysis showed that emodin decreased nuclear factor kappa B (NF-kappa B) expression in 24 h of treatment, but in 48 h, emodin increased NF-kappa B activity. A confocal microscope showed that emodin induced NF-kappa B translocation from cytoplasm to nuclei. C6 cells would activate the mitogen-activated protein kinase survival pathway and express the DNA repair gene (MGMT) and associated proteins (PARP and XRCC1) to recover the cell activity. C6 cells also expressed GRP78 to decrease emodin-induced endoplasmic reticulum (ER) stress that would cause apoptosis in C6 cells, and GRP78 inhibited the expression of GADD153 to enhance the expression of Bcl-2 that could balance the ER-and mitochondria-induced apoptosis of C6 cells

Beneficial effects of treatment with cystamine on brain in NZB/W F1 mice

[[abstract]]The involvement of the central nervous system (CNS) or neuropsychosis has been reported in patients with systemic lupus erythematosus (SLE) and considered a major cause of long-standing functional impairment and mortality. However, little is known in the improvement of the brain abnormality in SLE.To investigate the effect and mechanism of cystamine on brain in SLE, NZB/W F1 mice were used as the animal model. Gel zymography, caspase-3 activity assay and Western blots were performed to elucidate the effect of cystamine. Significant reduction of matrix metalloproteinases (MMP)-9/MMP-2 ratio and urokinase-type plasminogen activator (uPA) expression was detected in brain of NZB/W F1 mice treated with cystamine as compared to control group. Significant increase of heat-shock protein (HSP)-70 and HSP27 was detected in brain of NZB/W F1 mice treated with cystamine as compared to control group. Additionally, significant reduction of mitochondrial dependent apoptosis was observed in brain of NZB/W F1 mice treated with cystamine as compared to control group by increasing BCL-2 and reducing caspase-9, Bad, and Apaf-1 expression. Moreover, increased phosphorylated p65 (NF-kappa B) protein was observed in brain of NZB/W F1 mice treated with cystamine as compared to control group. These experimental results firstly demonstrated the beneficial effects of cystamine on brain in NZB/W F1 mice and suggested the therapeutic potential in patients with neuropsychiatric SLE (NP-SLE). (c) 2008 Elsevier B.V. All rights reserved

AmpD(I) Is Involved in Expression of the Chromosomal L1 and L2 beta-Lactamases of Stenotrophomonas maltophilia

[[abstract]]Two ampD homologues, ampD(I) and ampD(II), of Stenotrophomonas maltophilia have been cloned and analyzed. Comparative genomic analysis revealed that the genomic context of the ampD(II) genes is quite different, whereas that of the ampD(I) genes is more conserved in S. maltophilia strains. The ampD system of S. maltophilia is distinct from that of the Enterobacteriaceae and Pseudomonas aeruginosa in three respects. (i) AmpD(I) of S. maltophilia is not encoded in an ampDE operon, in contrast to what happens in the Enterobacteriaceae and P. aeruginosa. (ii) The AmpD systems of the Enterobacteriaceae and P. aeruginosa are generally involved in the regulation of ampR-linked ampC gene expression, while AmpD(I) of S. maltophilia is responsible for the regulation of two intrinsic beta-lactamase genes, of which the L2 gene, but not the L1 gene, is linked to ampR. (iii) S. maltophilia exhibits a one-step L1 and L2 gene derepression model involving ampD(I), distinct from the two-or three-step derepression of the Enterobacteriaceae and P. aeruginosa. Moreover, the ampD(I) and ampD(II) genes are constitutively expressed and not regulated by the inducer and AmpR protein, and the expression of ampD(II) is weaker than that of ampD(I). Finally, AmpD(II) is not associated with the derepression of beta-lactamases, and its role in S. maltophilia remains unclear

Transglutaminase inhibitor cystamine alleviates the abnormality in liver from NZB/W F1 mice

[[abstract]]Increased hepatic abnormality has been observed in patients with systemic lupus erythematosus (SLE) and contributes to the elevated apoptosis that results in severe disease activity. Since cystamine has been demonstrated to be beneficial for NZB/NV F1 mice, this study investigates the effects of cystamine on various inflammatory and stress-related proteins in liver from NZB/W F I mice. Nephelometric analyses and immunoblots were conducted to detect aspartate aminotransferase (AST), alanine aminotransferase (ALT), C-reactive protein (CRP), p53, p21, Gadd45, heat shock protein 70 (HSP70) and cyclooxygenase-2 (COX-2). AST and ALT were reduced in NZB/W F1 mice that were given cystamine and CRP, p53, p21, Gadd45, HSP70 and COX-2 proteins in the liver were reduced in NZB/W F1 mice that were treated with cystamine. Moreover, cystamine has no obvious effect on BALB/c mice. These findings suggest that cystamine reduces the inflammation in liver of NZB/W F1 mice and provide a clue in treatment of SLE with liver abnormality. (c) 2007 Elsevier B.V. All rights reserved

Involvement of reactive oxygen species and caspase-dependent pathway in berberine-induced cell cycle arrest and apoptosis in C6 rat glioma cells

[[abstract]]The cytotoxicity of berberine on C6 rat glioma cells indicated that berberine induced morphological changes and caused cell death through G2/M arrest and apoptosis. While undergoing apoptosis, there was a remarkable accumulation of G2/M cells with the upregulatoin of Weel but it also inhibited cyclin B, CDK1 and Cdc25c that led to G2/M arrest. Along with cytotoxicity in C6 cells, several apoptotic events including mitochondrial cytochrome c release, activation of caspase-9, -3 and -8 and DNA fragmentation were induced. Berberine increased the levels of GADD153 and GRP 78 in C6 cells based on the examination of Western blotting and this is a major hallmark of endoplasmic reticulum (ER) stress. We also found that berberine promoted the production of reactive oxygen species and Ca2+ in C6 cells. Western blotting assay also showed that berberine inhibited the levels of anti-apoptotic protein Bcl-2 but increased the levels of pro-apoptotic protein Bax before leading to a decrease in the levels of mitochondrial membrane potential (Delta psi(m)) followed by cytochrome c release that caused the activations of capase-9 and -3 for apoptotic occurrence. The caspase-8, -9 and -3 were activated by berberine in C6 cells based on the substrate solution (PhiPhiLux-G(1)D(1), CaspaLux 8-L1D2, CaspaLux 9-M1D2 for caspase-3, -8 and -9, respectively) and analyzed by flow cytometer and each inhibitor of caspase-8, -9 and -3 led to increase the percentage of viable C6 cells after exposure to berberine. This finding was also confirmed by Western blot assay which showed that berberine promoted the active form of caspase-8, -9 and -3. These results demonstrate that the cytotoxicity of berberine in C6 rat glioma cells is attributable to apoptosis mainly through induced G2/M-arrested cells, in an ER-dependent manner, via a mitochondria-dependent caspase pathway regulated by Bax and Bcl-2

Flagellar Biogenesis of Xanthomonas campestris Requires the Alternative Sigma Factors RpoN2 and FliA and Is Temporally Regulated by FlhA, FlhB, and FlgM

[[abstract]]In prokaryotes, flagellar biogenesis is a complicated process involving over 40 genes. The phytopathogen Xanthomonas campestris pv. campestris possesses a single polar flagellum, which is essential for the swimming motility. A sigma(54) activator, FleQ, has been shown to be required for the transcriptional activation of the flagellar type III secretion system (F-T3SS), rod, and hook proteins. One of the two rpoN genes, rpoN2, encoding sigma(54), is essential for flagellation. RpoN2 and FleQ direct the expression of a second alternative sigma FliA (sigma(28)) that is essential for the expression of the flagellin FliC. FlgM interacts with FliA and represses the FliA regulons. An flgM mutant overexpressing FliC generates a deformed flagellum and displays an abnormal motility. Mutation in the two structural genes of F-T3SS, flhA and flhB, suppresses the production of FliC. Furthermore, FliA protein levels are decreased in an flhB mutant. A mutant defective in flhA, but not flhB, exhibits a decreased infection rate. In conclusion, the flagellar biogenesis of Xanthomonas campestris requires alternative sigma factors RpoN2 and FliA and is temporally regulated by FlhA, FlhB, and FlgM

Acid-sensing ion channel 3 mediates peripheral anti-hyperalgesia effects of acupuncture in mice inflammatory pain

<p>Abstract</p> <p>Background</p> <p>Peripheral tissue inflammation initiates hyperalgesia accompanied by tissue acidosis, nociceptor activation, and inflammation mediators. Recent studies have suggested a significantly increased expression of acid-sensing ion channel 3 (ASIC3) in both carrageenan- and complete Freund's adjuvant (CFA)-induced inflammation. This study tested the hypothesis that acupuncture is curative for mechanical hyperalgesia induced by peripheral inflammation.</p> <p>Methods</p> <p>Here we used mechanical stimuli to assess behavioral responses in paw and muscle inflammation induced by carrageenan or CFA. We also used immunohistochemistry staining and western blot methodology to evaluate the expression of ASIC3 in dorsal root ganglion (DRG) neurons.</p> <p>Results</p> <p>In comparison with the control, the inflammation group showed significant mechanical hyperalgesia with both intraplantar carrageenan and CFA-induced inflammation. Interestingly, both carrageenan- and CFA-induced hyperalgesia were accompanied by ASIC3 up-regulation in DRG neurons. Furthermore, electroacupuncture (EA) at the ST36 rescued mechanical hyperalgesia through down-regulation of ASIC3 overexpression in both carrageenan- and CFA-induced inflammation.</p> <p>Conclusions</p> <p>In addition, electrical stimulation at the ST36 acupoint can relieve mechanical hyperalgesia by attenuating ASIC3 overexpression.</p

Optimising antimicrobial prescription in hospitals by introducing an antimicrobial stewardship programme in Hong Kong: Consensus statement

Objective. To discuss the implementation of an 'antimicrobial stewardship programme' as a means to improve the quality of antimicrobial use in a hospital setting in Hong Kong. Participants. Consensus working group on 'antimicrobial stewardship programme', The Scientific Committee on Infection Control, Centre for Health Protection, Department of Health, comprised 11 experts. The remit of the working group was to discuss the rationale and requirement for optimising antimicrobial prescriptions in hospitals by the introduction of an 'antimicrobial stewardship programme'. Evidence. PubMed articles, national and international guidelines, and abstracts of international meetings published between January 2000 and December 2004 on programmes for improving the use of antimicrobials in hospitals. Only English medical literature was reviewed. Consensus process. Data search was performed independently by three members of the working group. They met on three occasions before the meeting to discuss all collected articles. A final draft was circulated to the working group before a meeting on 3 January 2005. Five commonly asked questions about an 'antimicrobial stewardship programme' were selected for discussion by the participants. Published information on the rationale, components, outcome measures, advantages, and disadvantages of the programme was reviewed. Recent unpublished data from local studies of an 'antimicrobial stewardship programme' were also discussed. The timing, potential problems, and practical issues involved in the implementation of an 'antimicrobial stewardship programme' in Hong Kong were then considered. The consensus statement was circulated to and approved by all participants. Conclusion. The continuous indiscriminate and excessive use of antimicrobial agents promotes the emergence of antibiotic-resistant organisms. Antimicrobial resistance substantially raises already-rising health care costs and increases patient morbidity and mortality. Pattern of prescriptions in hospitals can be improved through the implementation of an 'antimicrobial stewardship programme'. A 'universal' and 'continuous' 'antimicrobial stewardship programme' should now be established in Hong Kong hospitals.published_or_final_versio

Household out-of-pocket medical expenditures and national health insurance in Taiwan: income and regional inequality

BACKGROUND: Unequal geographical distribution of medical care resources and insufficient healthcare coverage have been two long-standing problems with Taiwan's public health system. The implementation of National Health Insurance (NHI) attempted to mitigate the inequality in health care use. This study examines the degree to which Taiwan's National Health Insurance (NHI) has reduced out-of-pocket medical expenditures in households in different regions and varying levels of income. METHODS: Data used in this study were drawn from the 1994 and 1996 Surveys of Family Income and Expenditure. We pooled the data from 1994 and 1996 and included a year dummy variable (NHI), equal to 1 if the household data came from 1996 in order to assess the impact of NHI on household out-of-pocket medical care expenditures shortly after its implementation in 1995. RESULTS: An individual who was older, female, married, unemployed, better educated, richer, head of a larger family household, or living in the central and eastern areas was more likely to have greater household out-of-pocket medical expenditures. NHI was found to have effectively reduced household out-of-pocket medical expenditures by 23.08%, particularly for more affluent households. With the implementation of NHI, lower and middle income quintiles had smaller decreases in out-of-pocket medical expenditure. NHI was also found to have reduced household out-of-pocket medical expenditures more for households in eastern Taiwan. CONCLUSION: Although NHI was established to create free medical care for all, further effort is needed to reduce the medical costs for certain disadvantaged groups, particularly the poor and aborigines, if equality is to be achieved