targeting ras for anticancer drug discovery

... Apart from NO synthesis L-arginine may also serve as a substrate for glutamate formation and may also provide increased substrate for arginase (50) No significant changes in the activity of arginase ... to stay at normal conditions and were given normal diet for days and decapitated and the different brain regions (CC, CB and BS) were used for study It was reported by Ananth et al (23) that days ... induction of excitotoxicity and hence that period (5 days) was chosen for the reperfusion group Enzyme assays: Total NOS activity (all isoforms of NOS: nNOS, iNOS & eNOS) was estimated by the method of...

Ngày tải lên: 26/10/2012, 09:07

... probed for ROS, specifically whether or not the organelles undergo permeability transition pore (PTP) formation Among the many features accompanying mitochondrial permeability transition (for a ... mediated ROS formation through the respiratory chain; i.e both complex I and KGDHC are in equilibrium with the NAD(P)H ⁄ NAD(P)+ ratio, and therefore interdependent on each other concerning ROS formation ... and V Adam-Vizi 48 Huennekens FM, Basford RE & Gabrio BW (1955) An oxidase for reduced diphosphopyridine nucleotide J Biol Chem 213, 951–967 49 Gazaryan IG, Krasnikov BF, Ashby GA, Thorneley RN,...

Ngày tải lên: 07/03/2014, 12:20

... electroporated into Agrobacterium tumefaciens (strain LBA4404) for tobacco transformation with the leaf disk method, and the transformants were screened for kanamycin (100 mgÆL)1) resistance The transgenic ... as controls For inoculation with pathogenic fungi, F oxysporum f sp vasinfectum and P parasitica var nicotianae Tucker were cultured on potato dextrose agar (PDA) medium at 25° C for 15 days, ... used for northern blot analyses Enzyme activity assays Tobacco seedlings were treated with 200 mm NaCl and 15% poly(ethylene glycol) 6000 for days, and 0.5 g of leaves was then collected for SOD,...

Ngày tải lên: 14/03/2014, 23:20

... association For each strain, 12 lL of overnight culture diluted to 0.2 OD600nm units and four subsequent : dilutions were spotted on plates Growth was monitored after days for all plates except for diamide, ... Dr Shusuke Kuge for providing strains and plasmids We also thank Hugo Metz for technical assistance with the confocal laser scanning microscopy analysis and Lyndel Meinhardt for his comments ... sensitivity of deletion mutants for the regulators most frequently associated with oxidative stress response: Yap1, Skn7, Msn2 and Msn4 Single or double mutants for Yap1 and Skn7 were very sensitive...

Ngày tải lên: 16/03/2014, 14:20

... Cell growth was continued for 4–6 h The cells were harvested and lysis was performed by two cycles of a freeze-thaw protocol Cell debris was separated by centrifugation for 30 at 100 000 g The ... strictly conserved and two extreme conformations have been observed for this residue in the crystal structure of Anx24(Ca32) In the current structure, the conformation of the AB loop in the first ... loop of Anx(Gh1) adopt a conformation that might be suited for coordination of a calcium ion, although not in a canonical fashion However, there is no experimental proof for calcium binding in this...

Ngày tải lên: 17/03/2014, 03:20

... propidium iodide for min, fixed with methanol at )20 °C for min, permeabilized with 0.1% (v/v) Triton X-100 in NaCl/Pi for 15 min, and stained with Cy3-labeled tubulin antibody (Sigma) For assessment ... and C Versluis for performing MS measurements, and to C L H Guikers for assistance with PDT experiments References Dolmans, D.E., Fukumura, E & Jain, R.K (2003) Photodynamic therapy for cancer Nat ... Hypocrellin A was loaded into the cells in the culture medium for h Then the medium with photosensitizer was removed, and the cells were incubated for 15 with acetylTyr-Fluo (5 lM) in NaCl/Pi supplemented...

Ngày tải lên: 23/03/2014, 15:21

... were centrifuged at 100,000 × g for hr at 4°C Supernatants were removed, pellets were re-suspended in 70% formic acid and sonicated and centrifuged at 100,000 × g for hr at 4°C Supernatants were ... deparaffmized, hydrated, rinsed with PBS and pretreated with formic acid (88%) for 10 to antigen retrieval, and with 3% H202 in methanol for 30 to eliminate endogenous peroxidase activity in the ... intra-cardially for 30 with ice-cold 0.9% phosphate buffer saline (PBS), containing EDTA (2 mM/L) and BHT (2 mM/L), pH7.4 Brains were removed and one hemisphere was fixed by immersion in 4% paraformaldehyde...

Ngày tải lên: 19/06/2014, 22:20

... transforming growth factor Inter- and intra-assay variances for VEGF, sVEGFR-1, and sVCAM-1 were lower than 10% The minimum detectable concentration was less than pg/ml for VEGF, less than pg/ml for ... days before hospitalisation The baseline evaluation was performed on the morning of admission after hour of rest at room temperature The duration of the wash-out period is long enough for calcium-channel ... permeability-enhancing activities specific for endothelial cells It interacts with two receptor tyrosine kinases, VEGFR-1 (flt) and VEGFR-2 (flk) A defect in VEGF receptors could account for VEGF signalling abnormalities...

Ngày tải lên: 09/08/2014, 01:23

... primers for GLUT-1 and endogenous control genes were designed using Beacon Designer software (PREMIER Biosoft International, Palo Alto, CA, USA) Details of the forward and reverse primers for the ... Quantitative real-time RT-PCR was performed with iTaq™ DNA polymerase using iQ™ SYBR Green Supermix (BioRad Laboratories) The efficiency of the amplification reaction for each gene was calculated by ... the analogous values for affinity (1.07 ± 0.28 and 1.49 ± 0.59 mM, respectively) and maximal velocity (0.27 ± Page of 11 (page number not for citation purposes) To control for possible osmotic...

Ngày tải lên: 09/08/2014, 14:21

... 0.4 μM) of forward and reverse primers in a final volume of 20 μl PCR reactions were conducted as follows: initially at 50°C for min, 95°C for min, 40 cycles for denaturation at 95°C for 15 s, ... hemodialysis (HD) for more than months; showed no evidence of chronic or acute infections, inflammatory disorders, and malignancy; and were not using anti-inflammatory drugs for months before enrollment ... annealing at 60°C for 20 s, and primer extension at 72°C for 15 s, and finally at 25°C The threshold cycle number (Ct) values of the Chr1 gene and ND2 gene were determined for each individual...

Ngày tải lên: 11/08/2014, 07:21

... GAPDH Forward Reversed Forward Reversed Forward Reversed Forward Reversed Forward Reversed Forward Reversed Forward Reversed Forward Reversed Forward Reversed Forward Reversed Forward Reversed Forward ... 5-minute polymerase activation step and continued with for 40 cycles (denaturation) at 95°C for 20 sec, annealing for 30 sec, and elongation at 72°C for 30 sec with a final extension for at 72°C ... Forward Reversed Forward Reversed Forward Reversed Forward Reversed Forward Reversed Forward Reversed Forward Reversed Forward Reversed Forward Reversed TGT CCC CAC CCC CAA TGT ATC CTC CGA TGC CTG CTT...

Ngày tải lên: 13/08/2014, 13:20

... acclimatize to the environmental conditions for one week before experiments All animal handling procedures were approved by the Ethics Committee of the National Centre for Radiation Research and Technology, ... blood sample was separated and kept frozen for biochemical assays Bronchoalveolar lavage was performed and lungs were weighed and processed separately for biochemical and histological studies Lung ... [29] A phase II clinical study shows BA to be a promising treatment for idiopathic pulmonary fibrosis [30] BA may be beneficial for a range of fibrotic conditions through both anti-inflammatory...

Ngày tải lên: 13/08/2014, 14:20

... Médecine Hyperbare (Angers, France) for financial support, P Legras and J Roux for animal care, M Gonnet for NaHS conditioning, and Ph Lane, C Hoffmann and P Pothier for English proofreading Author ... comparisons were performed using the Dunnett’s test for the effect of time and with Bonferroni’s correction for the effects of treatment at specific times The MannWhitney test was used for inter-group ... to measure MAP and HR and for the induction of hemorrhagic shock The homolateral femoral vein was canulated for retransfusion of shed blood, for fluid maintenance and for bolus infusion (either...

Ngày tải lên: 13/08/2014, 21:21

... cytokinin-regulated transcriptional network, microarray analyses have been performed by different groups (Brenner et al 2005; Rashotte et al 2003; Rashotte et al 2006) In addition to the type B ARRs, there are ... Stresses 1.9 25 Brassinosteroids The presence of brassinosteroids (BRs) in almost all tissues of a plant and in almost every species of plant kingdom has been demonstrated BRs occur in free form and ... concentration Thus, an understanding of the phytohormones homeostasis and signaling is essential for improving plant performance under optimal and stressful environments v vi Preface Traditionally five major...

Ngày tải lên: 14/02/2014, 12:20

... on resealing For controls, BSA (1 mgÆmL)1) and lysozyme (1 mgÆmL)1) were used instead of a-crystallin The tubes containing the suspensions were placed on ice for 10 then at 37 °C for 30 so that ... ghost cells are in their open form, allowing various proteins to be incorporated before resealing SDS/PAGE analyses of ghost cells alone showed the usual pattern for red cell membrane proteins ... charged There is no active transport for methylglyoxal or H2O2 in the erythrocyte membrane There is an active glucose-transporter but it is specific for glucose and not for other hexoses; fructose can...

Ngày tải lên: 17/03/2014, 03:20

... Unfortunately, all gid mutants retained full induction of the GPXH wild-type gene (data not shown) For the 32 gox mutants, a secondary screen was performed to reduce the number of strains for ... intensity of several mutants indicates that the former might be the reason for high GPXH expression in some of the gox mutants The formation of 1O2 was therefore measured with the fluorescent dye singlet ... cells were dark adapted for 20 before maximal chlorophyll fluorescence level Fm was determined Then actinic light of the same intensity as growth light was turned on for 15 before F m ’ was measured...

Ngày tải lên: 11/08/2014, 11:21

... Luciferase Reporter Assays For Tat assays, 0.3 μg of the reporter AGP546 (TKp-TARluciferase) was co-transfected with 10 μg of the Tatexpression plasmid AGP535, and 0.5 μg of the CMV-GFP plasmid For ... FR-TKp-luciferase, in which a cluster of 20 EBNA1 binding sites from the EBV genome is placed 5' to a TKp-luciferase reporter cassette [33,39] We constructed an analogous reporter for Tat by inserting ... between TKp and the luciferase gene This reporter, TKp-TAR-luciferase, contains 77 nucleotides of HIV-1 sequence from the LAV strain of HIV-1 between the TKp and luciferase [35] The first nucleotide...

Ngày tải lên: 12/08/2014, 04:20

... monoclonal antibodies for surface staining for 15 in the dark at room temperature Red blood cells were ruptured with ml lysis solution (FACS lysing solution, BD Bioscience) for 10 in the dark at ... analyses were performed using the CellQuest software (BD Bioscience) in a FACSCalibur four-color flow cytometer (BD Bioscience) For each condition 5,000 events were counted in forward- and side-scatter ... followed by centrifugation (250 g for 10 at 25°C) Hypotonic lyses in 0.2% saline were performed followed by addition of 1.6% saline and centrifugation (250 g for 10 at 25°C) The supernatants were...

Ngày tải lên: 13/08/2014, 10:20

... National Orthopedic Hospital Enugu (NOHE), Nigeria for follow up treatment from September to December 2008 Consecutive injured RTA victims who gave informed consent were interviewed Patients with ... also be prone to developing post-traumatic syndrome and stress disorder For this reason, a second control group was considered for the study Thus, 150 individuals from staffs of Federal Neuropsychiatric ... injury following the RTAs and patients whose RTAs had not lasted for more than one month However, none of the patients approached for interview refused to give consent Patients with head injury...

Ngày tải lên: 13/08/2014, 23:20