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| 1 | Screening Helicobacter pylori genes induced during infection of mouse stomachs显示文摘AIM:To investigate the effect of in vivo environment on gene expression in Helicobacter pylori(H.pylori) as it relates to its survival in the host.METHODS:In vivo expression technology(IVET) systems are used to identify microbial virulence genes.We modified the IVET-transcriptional fusion vector,pIVET8,which uses antibiotic resistance as the basis for selection of candidate genes in host tissues to develop two unique IVET-promoter-screening vectors,pIVET11 and pIVET12.Our novel IVET systems were developed by the fusion of random Sau3A DNA fragments of H.pylori and a tandem-reporter system of chloramphenicol acetyltransferase and beta-galactosidase.Additionally,each vector contains a kanamycin resistance gene.We used a mouse macrophage cell line,RAW 264.7 and mice,as selective media to identify specific genes that H.pylori expresses in vivo.Gene expression studies were conducted by infecting RAW 264.7 cells with H.pylori.This was followed by real time polymerase chain reaction(PCR) analysis to determine the relative expression levels of in vivo induced genes.RESULTS:In this study,we have identified 31 in vivo induced(ivi) genes in the initial screens.These 31 genes belong to several functional gene families,including several well-known virulence factors that are expressed by the bacterium in infected mouse stomachs.Virulence factors,vacA and cagA,were found in this screen and are known to play important roles in H.pylori infection,colonization and pathogenesis.Their detection validates the efficacy of these screening systems.Some of the identified ivi genes have already been implicated to play an important role in the pathogenesis of H.pylori and other bacterial pathogens such as Escherichia coli and Vibrio cholerae.Transcription profiles of allivi genes were confirmed by real time PCR analysis of H.pylori RNA isolated from H.pylori infected RAW 264.7 macrophages.We compared the expression profile of H.pylori and RAW 264.7 coculture with that of H.pylori only.Some genes such as cag A,vac A,lpx C,mur I,tlp C,trx B,sod B,tnp B,pgi,rbf A and inf B showed a 2-20 fold upregulation.Statistically significant upregulation was obtained for all the above mentioned genes(P < 0.05).tlp C,cag A,vac A,sod B,rbf A,inf B,tnp B,lpx C and mur I were also significantly upregulated(P < 0.01).These data suggest a strong correlation between results obtained in vitro in the macrophage cell line and in the intact animal.CONCLUSION:The positive identification of these genes demonstrates that our IVET systems are powerful tools for studying H.pylori gene expression in the host environment. | Aparna Singh Nathaniel Hodgson Ming Yan Jungsoo Joo Lei Gu Hong Sang Emmalena Gregory-Bryson William G Wood Yisheng Ni Kimberly Smith Sharon H Jackson William G Coleman | 2012 | World Journal of Gastroenterology2012，18，32： | 5 |
| 2 | Coping style use predicts posttraumatic stress and complicated grief symptom severity among college students reporting a traumatic loss显示文摘 | Kimberly R S Jon D E Matt J G | 2007 | Journal of Counseling Psychology2007，54，3： | 1 |
| 3 | Failure of methotrexate and internaliliac balloon catherization to manage placenta percreta显示文摘 | Kimberly B Alain G Marie FD | 2002 | Obstet Gynecol2002，99，： | 1 |
| 4 | Musculoskeletal ultra- sound : taking sports medicine to the next level显示文摘 | Kimberly G Harmon Francis G O 'Connor | 2010 | Br J Sports Med2010，44，16： | 1 |
| 5 | 显示文摘 | Kevin L S Paul A D Kimberly A G | 2010 | Applied Catalysis B Environmental2010，97，34： | 1 |
| 6 | A primer on newborn screening显示文摘 | Kristin G B Kimberly M H Nicola L | 2004 | Advances in Neonatal Care2004，4，5： | 1 |
| 7 | Removel of algal-derived organic material by preozona- tion and coagulation:monitoring changes in or- ganic quality by Pyrolgsis - GC - MS 显示文摘 | David L W Kimberly A G Kim S M | 1996 | Wat Res1996，30，11： | 1 |
| 8 | Siegmund,et al,Prognostic relevance of methylation markers in patients with non-muscle invasive bladder carcinoma显示文摘 | Martin G Friedrich Shahin Chandrasoma Kimberly D | 2005 | Eur J Cancer2005，41，17： | 1 |
| 9 | 显示文摘 | Gibson V C Kimberly B S | 1998 | J Chem Cummun1998，7，： | 1 |
| 10 | Short-term tensile creep and shrinkage of ultra-high performance concrete显示文摘 | Victor Y G Lawrence F K Kimberly E K | 2009 | Cement and Concrete Composites2009，，3： | 1 |
| 11 | A reference method laboratory network for cholesterol:a model for standardization and improvement of clinical laboratory measurements显示文摘 | Kimberly M M Waymack P P | 2000 | Clin Chem2000，46，11： | 1 |
| 12 | Failure of methotrexate and internal iliac balloon catherization to manage placenta percreta显示文摘 | Kimberly B Alain G Marie FD | 2002 | Obstet Gynecol2002，99，： | 1 |
| 13 | Effects of music on physiological and affective responses to graded treadmill exercise in trained and untrained runners显示文摘 | Kimberly A Brownley Robert G McMurray Anthony C Hackney | 1995 | International Journal of Psychophysiology1995，19，： | 1 |
| 14 | Defining the Intellectual Profile of Asperger Syndrome: Comparison with High-Functioning Autism显示文摘 | Mohammad G Kimberly MK | 2004 | Journal of Autism and Developmental Disorders2004，3，34： | 1 |
| 15 | Effect of crystal phase composition on the reductive and oxidative abilities of TiO2 nanotubes under UV and visible light显示文摘 | KEVIN L S PAUL A D KIMBERLY A G | 2010 | Applied Catalysis B: Environmental2010，97，35： | 1 |
| 16 | Explaining the enhanced photocatalytic activity of degussa P25 mixed-phase TiO2 using EPR显示文摘 | DEANNA C HURUM ALEXANDER G AGRIOS KIMBERLY A GRAY | 2003 | J Phys Chem B2003，107，19： | 1 |
| 17 | Effectiveness of Multimodal Pain Management Protocol in Total Knee Arthroplasty Patients显示文摘 | Cynthia L Kathleen G Kimberly M | | 0，，03： | 1 |
| 18 | Preparation of Mixed-Phase Titanium Dioxide Nanocomposites via Solvothermal Processing显示文摘 | Li Gong-hu Kimberly A G | | 0，，05： | 1 |
| 19 | Redefining the properties of an osmotic agent in an intestinal-specific preservation solution显示文摘AIM: To investigate the effects of dextrans of various molecular weights (Mw) during a 12 h cold storage time-course on energetics, histology and mucosal infiltration of fluorescein isothiocyanate (FITC)-dextran. METHODS: Rodent intestines were isolated and received a standard University of Wisconsin vascular flush followed by intraluminal administration of a nutrientrich preservation solution containing dextrans of varying Mw: Group D1, 73 kdal; Group D2, 276 kdal; Group D3, 534 kdal; Group D4, 1185 kdal; Group D5, 2400 kdal. RESULTS: Using FITC-labeled dextrans, fluorescent micrographs demonstrated varying degrees of mucosal infiltration; lower Mw (groups D1-D3: 73-534 kdal) dextrans penetrated the mucosa as early as 2 h, whereas the largest dextran (D5: 2400 kdal) remained captive within the lumen and exhibited no permeability even after 12 h. After 12 h, median injury grades ranged from 6.5 to 7.5 in groups D1-D4 (73-1185 kdal) representing injury of the regenerative cryptal regions and submucosa; this was in contrast to group D5 (2400 kdal) which exhibited villus denudation (with intact crypts) corresponding to a median injury grade of 4 (P < 0.05). Analysis of tissue energetics reflected a strong positive correlation between Mw and adenosine triphosphate (r 2 = 0.809), total adenylates (r 2 = 0.865) and energy charge (r 2 = 0.667).CONCLUSION: Our data indicate that dextrans of Mw > 2400 kdal act as true impermeant agents during 12 h ischemic storage when incorporated into an intraluminal preservation solution. | Kimberly Schlachter Matthew S Kokotilo Jodi Carter Aducio Thiesen Angela Ochs Rachel G Khadaroo Thomas A Churchill | 2010 | World Journal of Gastroenterology2010，16，45： | 1 |
| 20 | Evaluation of Dry and Wet Transported Intravaginal Swabs in Detection of Chlamydia trachomatis and Neisseria gonorrhoeae Infections in Female Soldiers by PCR显示文摘 | Charlotte A G Kimberly A C Nina S | 2002 | J Clin Microbiol2002，40，3： | 1 |
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