culture cell isolation rna extraction and rt pcr

... (Millipore, Bedford, MA) to remove bacteria and particles, and serial dilutions 1:10 were made Reverse Transcription (RT) -PCR RNA isolation, cDNA synthesis and RTPCR were performed as previously described ... http://respiratory-research.com/content/8/1/84 Cell lines and culture Human airway epithelial cells (A549, European Collection of Cell Cultures, Porton Down, UK) were cultured at 37°C in 5% CO2 in Ham's F12 (Gibco-BRL), ... collection and analysis, gene and protein expression analysis, drafted the manuscript and contributed to study design and analysis CG carried out LSC, cell viability studies and immunoassays, and contributed...

Ngày tải lên: 12/08/2014, 15:21

... microanalysis and water content measurement FT performed ELISA and RTPCR for cytokine analysis CK participated in cell culture and immunocytochemistry BNR performed western blot analysis JM, GB, MJ, CC and ... supernatant on cell viability To assess the effect of S aureus supernatant on cell viability, airway glandular cells were incubated with increasing concentrations of bacterial supernatant Cell ... quantitative RT- PCR analysis of IL-8 and TNFa mRNA levels The incubation with S aureus supernatant for h increased significantly (*, p < 0.05) IL-8 and TNFa mRNA levels compared to control condition (A and...

Ngày tải lên: 12/08/2014, 14:20

... http://respiratory-research.com/content/8/1/19 the alternative medication and started the next arm of the study Two study arms compared the bronchodilator response to salbutamol after formoterol and placebo treatment and the other two ... formoterol and theoretically it may be less likely to induce tolerance Alternatively, salmeterol could act as a partial antagonist to salbutamol and reduce the effects of salbutamol still further ... where airway inflammation, oedema and mucus production play important roles in airway obstruction Nevertheless, airway smooth muscle contraction is important and is the main target of emergency...

Ngày tải lên: 12/08/2014, 15:20

... Woodcock has received support for research, and honoraria for lectures and consultations, from AstraZeneca, Chiesi, GlaxoSmithKline, Novartis, Oriel Therapeutics, Novartis and Schering-Plough Acknowledgements ... products [9,10], and the data imply some different clinical profiles due to the different pharmacological properties of the ICS and LABAs [9,11-13] When taken on a regular basis, both SFC and FBC have ... covariance, which included terms for subject, treatment, period and baseline, and was used to estimate the treatment difference and p-value and to calculate the 95% confidence interval for this difference...

Ngày tải lên: 12/08/2014, 16:20

... fluid transport in cultured human alveolar epithelial type II cells Am J Physiol Lung Cell Mol Physiol 2006, 290:242-249 Jiang X, Ingbar DH, O’Grady SM: Adrenergic stimulation of Natransport across ... beta-agonists might improve fluid clearance and thus support the findings of our study [1] and the beta-agonist lung injury trial [2] Hopefully future studies will include randomized controlled trials to ... across alveolar epithelial cells involves activation of apical Cl-channels Am J Physiol Cell Physiol 1998, 275:16101620 Eisenhut M, Wallace H, Barton P, Gaillard E, Newland P, Diver M, Southern...

Ngày tải lên: 12/08/2014, 23:23

... minutes The selected ions monitoring mode is performed at m/z 227, 300, 335, 336 and 337 for CLEN, and 207, 294, 350, 369 and 370 for SAL Recoveries of CLEN, SAL 65% 88% The limits of detection were ... mẫu cột Oasis HLB 1cc/30mg Extraction Cartridge Đầu dò: Fluorescence - Phương pháp phân tích SAL CLEN sắc ký lỏng ghép khối phổ[34] Xử lý mẫu: Mẫu clean up cột MCX extraction plate, 10mg/96-well ... lần (Hamilton) lọc ion MultiPure RO (Fistreem) - Tủ sấy Memmert - Bồn siêu âm Minor- decon - Máy vortex Rotalab (OSI) - Cân phân tích Sartorius (chính xác đến 0,0001g) - Cột SPE 500mg EV SCX Alltech,...

Ngày tải lên: 08/02/2015, 21:54

... Journal 276 (2009) 6603–6614 ª 2009 The Authors Journal compilation ª 2009 FEBS 6607 Fibroblast proliferation and collagen synthesis A WO siRNA Control siRNA W Han et al A β-catenin siRNA WO siRNA ... D2 Con D1 D2 W O siRNA Control siRNA β-catenin siRNA 0.1 C Proliferation (absorbance) C 0.5 0.4 1.25 α-defensin-1 (μM) WO siRNA Control siRNA β-catenin siRNA * * 2.5 * COL1A1 mRNA level (2–ΔΔCT) ... dephosphorylated b-catenin, and collagen-I (Figs 7A–C and 8A–C) Moreover, a-defensin-induced increases in cell proliferation were prevented by SB203580 and LY294002 (Figs 7D and 8D) FEBS Journal 276 (2009)...

Ngày tải lên: 18/02/2014, 06:20

... experiments, and drafted the manuscript H-RS carried out the preparation of cells and the treatment of antibody and fluorescence K-BS conceived the basic idea of the whole experiment and supported the ... biofluidic part where the cultured cells are introduced on top of the optical filter through a polydimethylsiloxane [PDMS] channel and an underlying thin film p-FET as the signal transducer part which ... wavelength (λem) is transmitted through the filter and converted to the electrical signal The inset shows the actual p-FET device Figure Cell culture and visualization on the p-FET device (a) A microscopic...

Ngày tải lên: 20/06/2014, 20:20

... subsequent changes in the mRNA level of IL-17R and IL-17RB were assessed by RT- PCR 0, 1, 3, 6, 9, and 24 hours after the start of in vitro culture The relative intensity of each PCR band was normalized ... 20 ng/ml), and subsequent changes in the mRNA levels of IL-17R and IL-17RB were assessed by RT- PCR at hours after the onset of in vitro culture The relative intensity of each PCR band was normalized ... U/ml penicillin, and 100 µg/ml streptomycin Suspended cells were plated in 75-cm2 culture flasks and cultured at 37°C in 5% CO2 Medium was replaced every days, and once the primary culture reached...

Ngày tải lên: 09/08/2014, 01:23

... HR represents heart rate and bpm represents beat per minute B: Heart rate ten minutes after successful re-beat HR represents heart rate and bpm represents beat per minute The aortic-cross clamping ... the cardiac recovery and the heart oxygen delivery and consumption balance, which increased the myocardial energy stores and thereby benefit for the weaning process More and more physicians incline ... alignment and drafted the manuscript Second author: DZ participated in the design of the study and performed the statistical analysis Correspondence author: QY conceived of the study, and participated...

Ngày tải lên: 10/08/2014, 09:22

... μg RNA was Lee et al Journal of Inflammation 2011, 8:16 http://www.journal-inflammation.com/content/8/1/16 Flow cytometry RAW 264.7 cells (1 X 106 cells/ ml) were cultured in Petri-dishes The cells ... inhibited PGE2 production (Figure 2B) and mRNA of COX-2 in RAW 264.7 cells (Figure 2C) and primary macrophages (Figure 2D) by western blot analyses and subsequent RT- PCR NO is known to be a pro-inflammatory ... by multiple cell types involved in antigen presentation Both B7-1 and B7-2 are constitutively expressed on dendritic cells and are regulated on monocytes, macrophages, B cells, and T cells following...

Ngày tải lên: 11/08/2014, 03:20

... bEND.3 cells, cocultures of bEND.3 cells with BV2 cells led to LPS induced injury to bEND.3 cells (Figure 7A-C) and NO accumulation (Figure 7D) This toxic effect seemed to require cell- cell interactions, ... atmosphere and at 37°C, cells were plated in 75 cm2 cell culture flask (Corning, Acton, MA, USA) and were split twice a week For the experiments, cells were plated on 6-well dishes (1-2 × 106cells/well) ... increase endothelial cell death due to LPS, reversal by NOS and ROS inhibitors While LPS did not affect bEND.3 cells alone, when cultured with BV2 cells, LPS increased cell death and monolayer disruption...

Ngày tải lên: 11/08/2014, 06:22

... μg RNA was Lee et al Journal of Inflammation 2011, 8:16 http://www.journal-inflammation.com/content/8/1/16 Flow cytometry RAW 264.7 cells (1 X 106 cells/ ml) were cultured in Petri-dishes The cells ... inhibited PGE2 production (Figure 2B) and mRNA of COX-2 in RAW 264.7 cells (Figure 2C) and primary macrophages (Figure 2D) by western blot analyses and subsequent RT- PCR NO is known to be a pro-inflammatory ... by multiple cell types involved in antigen presentation Both B7-1 and B7-2 are constitutively expressed on dendritic cells and are regulated on monocytes, macrophages, B cells, and T cells following...

Ngày tải lên: 11/08/2014, 06:23

... antibodies to the cell- adhesion molecule ICAM-1, the goblet cell marker Muc-5A, and the Th2 marker T1/ST2 (Santa Cruz BioTech, Santa Cruz, CA) Isolation of spleen cells and intracellular cytokine ... increased the number of cells producing IL-10 and Sal increased cells producing IL-4, whereas FPS decreased the number of cells producing IL-10, IL-4 and IFN-γ Lung epithelial cells respond to inflammatory ... sacrificed and single -cell suspensions were prepared from the spleens and cultured for 24 to 48 hrs in DMEM with 10% FBS Splenocytes were incubated on anti-CD3-coated or OVA-coated plates for 24 h and...

Ngày tải lên: 20/06/2014, 01:20

... Jolla, CA, USA) and Martin Eilers (IMT Marburg, Germany) for plasmid vectors, and to Margitta Alt and Bernard Wilke for their excellent technical assistance This work was supported by grants from ... histidinetagged ubiquitin (Ubi-His; lanes and 4) The cells were either treated with GW501516 (lanes and 4) or left untreated (lanes and 3) Cells were harvested after 24 h and analyzed by immunoblotting ... protein levels and GW501516 on oligomerization of PPARb (A) Co-immunoprecipitation of FLAP-PPARb and PPARb HEK293 cells were cotransfected with pCDNA3.1zeo-3xFlag-mPPARb and pCMX-mPPARb Cells were...

Ngày tải lên: 30/03/2014, 03:20

... both memory and naïve B cells as defined by the presence or absence of CD27 expression We studied the effect of CP-870,893 on B cell activation and B cell stimulation of T cells, and we analyzed ... B cells and CD27+ memory B cells Cytokine secretion by in vitro stimulated B cells To determine whether CP-870,893 induces human B cells to produce cytokines, supernatant from stimulated B cells ... irradiated, then co-cultured for days with CFSE-labeled purified allogeneic CD4+ T cells at the indicated B cell: T cell titrations (A) Percentage of CFSElow T cells and (B) T cell IFN-gamma production...

Ngày tải lên: 18/06/2014, 15:20

... outcomes were days alive and free of PaO2/ FiO2

Ngày tải lên: 12/08/2014, 23:21

... ppb Part per billion (một phần tỉ) ppm Part per million (một phần triệu) RSD% Độ lệch chuẩn mẫu tương đối theo phần trăm SPE Solid Phase Extraction (Chiết pha rắn) SPE-SCX Solid Phase Extraction ... Hóa học Châu Á lần thứ 12, tháng năm 2007 Kuala Lumpur, Malaysia - International ```Symposium on Food Analysis, Chemistry and Technology, 12th Asian Chemical Congress, August 23-25, 2007, Kuala ... 3.3 Kết khảo sát giới hạn phát hiện, giới hạn định lượng phương pháp (Limit of Detection - LOD and Limit of Quantification -LOQ) 3.4 Độ dao động tỷ lệ cường độ mảnh ion 3.5 Tóm tắt qui...

Ngày tải lên: 26/08/2015, 13:06