extraction reverse transcription and quantitative real time polymerase chain reaction analysis
Báo cáo y học: "Tetra-O-methyl nordihydroguaiaretic acid (Terameprocol) inhibits the NF-B-dependent transcription of TNF-a and MCP-1/CCL2 genes by preventing RelA from binding its cognate sites on DNA" ppsx
... MA) RNA (1 μg) was denatured and reverse transcription was performed with the Improm ll reverse transcription kit (Promega, Madison, WI) in a reaction mix containing random hexamers as primers (50 ... at 45°C and DNA was purified using Qiaquick nucleotide removal columns (Qiagen) according to the manufacturer’s instructions ChIP Q-PCR and data analysis For realtime PCR, bound (3 μl) and input ... for TLR-3 and TLR-8 are double and single stranded RNA, respectively In these experiments we used the artificial ligands poly(I:C) for TLR-3 and resiquimod (R-848) for TLR-8 HEK293/TLR3 and HEK293/TLR8...
- 11
- 622
- 0
Tài liệu Báo cáo khoa học: Solution NMR structure of five representative glycosylated polyene macrolide antibiotics with a sterol-dependent antifungal activity doc
... nystatin A1 and rimocidin are summarized in Table Due to some spectrally degenerate proton resonances in the polyene and the polyol regions, complete extraction of the J coupling constants and the ... calculated, 21, 25, 29, 25 and 29 were retained, respectively, for pimaricin, nystatin A1, rimocidin, candidin and vacidin A on the basis of their low experimental and nonexperimental potential ... C28 and C29 at the junction between the tetraene C20–C27 and the diene C30–C33 The absence of experimental restraints in these two regions and the conformational space allowed to the C2–C10 and...
- 9
- 522
- 0
Tài liệu Báo cáo Y học: Ornithine decarboxylase-antizyme is rapidly degraded through a mechanism that requires functional ubiquitin-dependent proteolytic activity pot
... treated with cycloheximide and MG132 for the indicated times Cellular extracts were prepared, fractionated and antizyme and ODC detected as described in Fig Ó FEBS 2002 1320 S Gandre et al (Eur J Biochem ... growth medium of the cells and cellular extracts were prepared at various times thereafter The extracts were resolved by SDS/PAGE and antizyme was detected by Western blot analysis using specific ... °C) and cellular extracts were prepared 16 h thereafter The level of the indicated proteins was determined by Western blot analysis Western blot analysis Cells were harvested at the indicated times,...
- 7
- 382
- 0
Báo cáo khoa học: Characterization of the bga1-encoded glycoside hydrolase family 35 b-galactosidase of Hypocrea jecorina with galacto-b-D-galactanase activity pdf
... temperature and pH for catalysis The optimal temperature and pH for the reaction of Bga1 with oNPG as substrate was determined (Fig 4): maximal activity was found at pH 5.0 and 60 °C, and a substantial ... substrates and peptides and proteins for MS calibration were purchased from Sigma (St Louis, MO) and at least of analytical grade Arabinogalactan was purchased from Fluka (Buchs, Switzerland) and galactan ... confirmed that the enzyme released d-galactose in a time- and concentration-dependent manner The reaction followed a Michaelis–Menten kinetic, and the constants for the polymeric substrates are...
- 10
- 351
- 0
Báo cáo khoa học: Lending a helping hand, screening chemical libraries for compounds that enhance b-hexosaminidase A activity in GM2 gangliosidosis cells pptx
... of the same natural and artificial substrates as Hex A: Hex B (bb) is very thermostable and has a basic pI, and Hex S (aa) is thermolabile and has an acidic pI Only the Hex A and B isozymes are ... PYR-responsive (green) and nonresponsive (red) mutations in the a- (pink) and b- (blue) subunits of Hex A are labeled and drawn as spheres NGT (orange spheres) is shown bound in the a- and b-active sites ... Tay-Sachs or Sandhoff disease, respectively The majority of HEXA and HEXB mutations prevent any Hex A from being formed and result in the infantile ⁄ acute form of TaySachs (ITSD) or Sandhoff (ISD)...
- 11
- 348
- 0
Báo cáo khoa học: Selection of peptides inhibiting a b-lactamase-like activity docx
... °C for 10 and 35 cycles of denaturation at 94 °C for min, AB 348 and AB 349 primers annealing at 54 °C for and elongation at 72 °C for 1.5 Taq DNA polymerase was purchased from New England Biolabs ... after the negative selection (rounds NS and NS 2), i.e with IgG 9G4H9penicillin G covalent complex as a target Surface plasmon resonance (SPR) analysis SPR analysis is performed with a BIAcore X ... chips and running buffer HBS pH 7.4 containing 10 mM Hepes, 0.15 M NaCl, 3.4 mM EDTA and 0.005% surfactant P20, were from Pharmacia (Uppsala, Sweden) Immobilization of the biotinylated ligands...
- 7
- 428
- 0
Báo cáo y học: "Increased interleukin-17 production via a phosphoinositide 3-kinase/Akt and nuclear factor κB-dependent pathway in patients with rheumatoid arthritis" ppt
... for reverse transcription polymerase chain reaction as described PCR reaction product (25 µl) was separated on an agarose gel containing ethidium bromide The relative intensities of the bands ... whether enhanced IL-17 production could be regulated at a transcriptional level, semi-quantatitive reverse transcription polymerase chain reaction was performed R143 Arthritis Research & Therapy ... determination of IL-17 in test samples Quantification of IL-17 mRNA by semiquantitative reverse transcription polymerase chain reaction PBMC were incubated with various concentrations of antiCD3 in...
- 10
- 346
- 0
Báo cáo y học: "Tumor necrosis factor-alpha promotes survival in methotrexate-exposed macrophages by an NF-B-dependent pathwa" pptx
... experiments B-F PAR and BAY (10 mM or 7.5 mM) prevent TNF-a from rescuing MTX-treated RAW264.7 cells (B and C) and BMDM (D-F) from apoptosis Cells were exposed to PAR (B, D, and E) or BAY (C and F) for ... antirabbit and anti-mouse were used at 1:8,000 and 1:20,000 dilutions, respectively All antibodies were prepared in SignalBoost Immunoreaction Enhancer (Calbiochem, Darmstadt, Germany) and applied ... (Figure 1B) and annexin-V binding (Figure 1B) and significantly suppressed the sub-G0 (apoptotic) fraction on flow cytometric cell cycle analysis of MTX-exposed BMDM (Table 1) TNF-a did not reverse...
- 13
- 511
- 0
Tài liệu Báo cáo khoa học: Functional characterization of an orphan cupin protein from Burkholderia xenovorans reveals a mononuclear nonheme Fe2+-dependent oxygenase that cleaves b-diketones ppt
... six-coordinate and had two nitrogen(oxygen) donor ligands at an average distance of ˚ 1.90 A and four nitrogen(oxygen) ligands at an ˚ average distance of 2.06 A Three of the four ligands were reported ... nitrogen atoms of His60, His62 and His102 and a carboxylate oxygen atom, presumably contributed by the side chain of Glu96, are suggested to function as protein-derived ligands of the bound metal Mechanistic ... data analysis N is the number of ligands, r is the distance to the central iron atom, and r2 is the Debye–Waller factor The k-range is ˚ 2–13 A)1 See Table S1 for further details of EXAFS data analysis...
- 15
- 624
- 0
Tài liệu Báo cáo khoa học: Regulation of cathepsin B activity by 2A2 monoclonal antibody docx
... cathepsin B protein and activity are found in tumour tissues and have been suggested as prognostic markers in patients with breast, lung, colon and ovarian carcinomas, as well as gliomas and melanomas ... binding of substrates and other inhibitors and to identify the mechanism by which it regulates the activity of cathepsin B Results Preparation and characterization of 2A2 mAb and its Fab fragments ... 2A2 mAb and its Fab fragment (A) SDS ⁄ PAGE of the Fab fragment (lane 2); low molecular weight standards (lane 1) (B) IEF of 2A2 mAb (lane 2); IEF standards (lane 1) Friguet et al [30] and was...
- 13
- 417
- 0
Tài liệu Báo cáo khoa học: Dual P2Y12 receptor signaling in thrombin-stimulated platelets – involvement of phosphoinositide 3-kinase b but not c isoform in Ca2+ mobilization and procoagulant activity pdf
... Netherlands Organization for Scientific Research (902-16-276) and the Netherlands Heart Foundation (2002B014) 14 References Falati S, Gross P, Merrill-Skoloff G, Furie BC & Furie B (2002) Real- time ... relies on both phospholipase C and PI3-K activity PI3-Kb and not PI3-Kc mediates the P2Y12 effect on thrombin-evoked Ca2+ responses In man and mouse, the PI3-Kb (p110b) and PI3-Kc (p110c) isoforms ... al tirofiban from Merck, Sharp and Dohme (Haarlem, the Netherlands); Akt inhibitor and LY294002 from Calbiochem (La Jolla, CA, USA) ET-18-OCH3, manoalide, U73122 and U73343 came from Biomol (Plymouth...
- 15
- 565
- 0
Tài liệu Báo cáo khoa học: Glycation of low-density lipoprotein results in the time-dependent accumulation of cholesteryl esters and apolipoprotein B-100 protein in primary human monocyte-derived macrophages docx
... in a time- dependent manner, via scavenger receptor SR-A- and CD36-mediated processes, and that the uptake of lipid and protein occurs in synchrony Furthermore, it is shown that both lipid and ... fresh medium and LDL added at 48 h) before extraction and analysis by HPLC with UV detection Values are means ± SEM from three or more experiments, each with triplicate samples *, # and + indicate ... (e.g Ne-carboxymethyl-lysines and Ne-carboxyethyl-lysines and pentosidine) being known to accumulate with age on tissue proteins, and at an increased rate in LDL and atherosclerotic lesions in...
- 12
- 604
- 0
Tài liệu Báo cáo khoa học: Design, structure and biological activity of b-turn peptides of CD2 protein for inhibition of T-cell adhesion ppt
... as strong, medium and weak with upper and lower boundaries of distance for dNa (i, i), daN (i, i +1) and dNN (i, i +1), 1.9–3.0, 2.2–3.6 and ˚ 3.0–5.0 A, respectively [23] Side chain protons were ... hindrance between peptide and protein, and to relax the hydrogen added to the protein, the peptide and protein were merged and minimized before docking Atomic solvation parameters and fragmental volumes ... the interface of hCD2–hCD58 Comparison of hCD2 and rat CD2 3D structures indicated that Tyr86 and Glu36 side chains overlap in hCD2 and rat CD2 and oriented in the same direction in both the...
- 14
- 657
- 0
Tài liệu Báo cáo khoa học: The Alzheimer b-peptide shows temperature-dependent transitions between left-handed 31-helix, b-strand and random coil secondary structures doc
... PII-helix and PII has a smaller RH than an extended b-strand Figure shows the calculated hydrodynamic radius dependence on number of residues for a polypeptide chain in ideal random coil, b-strand and ... coil Val18, Phe19, Phe20 and Val24 on the other hand seem to start in a b-strand-rich conformation already at C, and go directly from there to random coil This high b-strand propensity is in good ... earlier often interpreted as random coil spectra However, no positive bands or local maxima should appear in a true random coil CD spectrum [22] The dihedral angles / and w that dene the PII structure...
- 12
- 287
- 0
Tài liệu Báo cáo khoa học: Molecular characterization and allergenic activity of Lyc e 2 (b-fructofuranosidase), a glycosylated allergen of tomato pdf
... some sera to a 9- and a 15-kDa band We could show that the 9-kDa band in the tomato extracts reacts with a specific antibody against the LTP from cherry, Pru av and the 15-kDa band shows reactivity ... desorption/ionization time- of-flight mass spectrometry (MALDI-TOF-MS) Further preparation and mass spectrometry analysis of N-glycans was performed according to Kolarich and Altmann [29] Briefly, ... reversed phase and an AG 3-X4A (Dow Chemical Company, Edegem, Belgium) Analysis and identification of the glycans was carried out by mass spectrometry using a DYNAMO MALDI-TOF (Thermo´ BioAnalysis,...
- 11
- 533
- 0
Tài liệu Báo cáo Y học: Characterization and regulation of yeast Ca2+-dependent phosphatidylethanolamine-phospholipase D activity docx
... the cytosolic and membrane-bound forms of yeast PtdEtn-PLD and examined the regulation of PtdEtn-PLD activity under certain growth, nutritional and stress conditions MATERIALS AND METHODS Chemicals ... (I+) and/ or mM choline (C+) Other amino acidrich media included: YPD [yeast extract and Bactopeptone (YP) containing 2% dextrose]; YPA (YP containing 0.05% glucose and 2% potassium acetate); and ... was collected and ultracentrifuged at 100 000 g for 90 The supernatant (cytosol) was collected and the resultant pellet (total membranes) was washed as above and resuspended in salt extraction buffer...
- 10
- 499
- 0
Báo cáo khoa học: The N-terminal hybrid binding domain of RNase HI from Thermotoga maritima is important for substrate binding and Mg2+-dependent activity pot
... Primers and 3, primers and 3, and primers and were used to amplify the genes encoding Tma-RNase HI, Tma-CD and Tma-ND, respectively The resultant DNA fragments were digested with NdeI and HindIII ... between the intact and truncated proteins and is calculated as: Tm (truncated) ) Tm (intact) DHm and DSm are the enthalpy and entropy changes of unfolding at Tm calculated by van’t Hoff analysis contrast, ... three-stranded anti-parallel b-sheet (b1–b3) and two helices (aA and aB) [22] It binds to the minor groove of the substrate, such that a loop between aA and b3 interacts with the RNA backbone and...
- 16
- 459
- 0
Báo cáo khoa học: Autocatalytic processing of procathepsin B is triggered by proenzyme activity doc
... procathepsin B in the presence of (s), 1.7 (d), 8.5 (h), 17 ( ) and 34 (D) nM E-64 at pH 4.5 and 37 °C Aliquots were taken from the reaction mixtures and added to 10 lM Z-Arg-Arg-NH-MEC substrate solution ... was obtained from Roche (Basel, Switzerland); polynucleotide T4 kinase was obtained from MBI Fermentas; and Vent DNA polymerase was obtained from New England Biolabs Oligonucleotides were obtained ... T58A, deletion of E59 and D60) as a template and mutagenic primers: 5¢-CCAGAACGTTCCGTTTGCACTGAA-3¢ (for T58ADED_M56P mutant: R54N, M56P, T58A, deletion of E59 and D60) and 5¢-GAACGTTATGCCGGCACTGA...
- 9
- 425
- 0
Báo cáo khoa học: Analysis of the NADH-dependent retinaldehyde reductase activity of amphioxus retinol dehydrogenase enzymes enhances our understanding of the evolution of the retinol dehydrogenase family pot
... encoding HA-RDH1 and HA-RDH2, respectively, and examined using confocal microscopy (D) Western blot of the pellets after 13 000 g (lanes and 4) and 100 000 g centrifugations (lanes and 3, microsomal ... similar-RDH2, rat similar-RDH1 and similarRDH2, and mouse truncated RDH and similar-RDH sequences were excluded from the phylogenetic analysis, as they were pseudogenes and ⁄ or partial RDH sequences ... and a structure–activity analysis of microsomal retinol dehydrogenases J Biol Chem 276, 19253–19258 Belyaeva OV & Kedishvili NY (2006) Comparative genomic and phylogenetic analysis of short-chain...
- 14
- 477
- 0
Tìm thêm: hệ việt nam nhật bản và sức hấp dẫn của tiếng nhật tại việt nam xác định các mục tiêu của chương trình xác định các nguyên tắc biên soạn khảo sát chương trình đào tạo của các đơn vị đào tạo tại nhật bản tiến hành xây dựng chương trình đào tạo dành cho đối tượng không chuyên ngữ tại việt nam điều tra đối với đối tượng giảng viên và đối tượng quản lí khảo sát thực tế giảng dạy tiếng nhật không chuyên ngữ tại việt nam khảo sát các chương trình đào tạo theo những bộ giáo trình tiêu biểu nội dung cụ thể cho từng kĩ năng ở từng cấp độ phát huy những thành tựu công nghệ mới nhất được áp dụng vào công tác dạy và học ngoại ngữ mở máy động cơ rôto dây quấn hệ số công suất cosp fi p2 đặc tuyến hiệu suất h fi p2 đặc tuyến mômen quay m fi p2 đặc tuyến tốc độ rôto n fi p2 đặc tuyến dòng điện stato i1 fi p2 động cơ điện không đồng bộ một pha thông tin liên lạc và các dịch vụ phần 3 giới thiệu nguyên liệu từ bảng 3 1 ta thấy ngoài hai thành phần chủ yếu và chiếm tỷ lệ cao nhất là tinh bột và cacbonhydrat trong hạt gạo tẻ còn chứa đường cellulose hemicellulose