|Resorufin - a lead for a new protein kinase CK2 inhibitor |
Iben Skjøth Sandholt1, Birgitte Brinkmann Olsen1, Barbara Guerra1, Olaf-Georg Issinger1, Brigitte Boldyreff2
Screening of a natural compound library led to the identification of resorufin, as a highly selective and potent inhibitor for protein kinase CK2. Out of 52 kinases tested only CK2 was inhibited. The IC50 values determined for the CK2 holoenzymes were 1.5 µM and for the free catalytic subunits ca. 4 µM. In different cancer cell lines treatment with resorufin led to cell death and endogenous CK2 was inhibited.
|Automation of cell proliferation assay sulforhodamine B fot the screening of stilbene derivatives|
Anne Soikkeli, Cristina Sempio, Erkki Mannila, Arto Urtti, Jouni Hirvonen, Marjo Yliperttula
The SRB assay was implemented on a TECAN Genesis workstation by optimizing cell numbers and liquid handling parameters. Anticancer drugs vinblastine, doxorubicin and 5-fluorouracil were used for standardizing the assay with a DU-145 cell line. To screen the antiproliferative activities of a series of stilbene derivatives, altogether 121 different stilbene structures, a plate layout was designed on 96-well format.
|Experimental Determination of ADMET Parameters in High Throughput, Using Colloidal Gold Composites and a Conductive Polymer as Reporting Reagents|
Roberto Martinez-Neira, Patricia De Pril, Anne Van Hoonacker, Patrick Englebienne
We developed and validated a high throughput in vitro setting to experimentally determine hSA and Orosomucoid affinities, fraction absorbed, Log BBB and Redox potential of NCEs using gold nanoparticles functionalized with proteins, lipids or conductive polymers. Such automated ADME assays provide means for more objective decisions in early drug discovery.
|High Throughput Anchorage-Independent Cell Colony Growth Assay with the IsoCyte™-HTS Platform|
Steven C. Miller1, Charlyn Sarnecki2, Paul B. Comita1, and Evan F. Cromwell1
The advantages of anchorage-independent cell colony growth assays for drug development are well known. Monitoring proliferation of cells in colony form is more biologically relevant than as single cells and formation of 3-dimensional (3D) colonies in semi-solid media better represents
in vivo environments.
|Treatment of Plaque Infections and Periodontitis using Stem Cells- A New Prospect|
Manoj Hariharan , Naveen Raj
Dental precursor cells are attractive for novel approaches to treat diseases like periodontitis, dental caries or to improve dental pulp healing and the regeneration of craniofacial bone and teeth. These cells are easily accessible and, in contrast to bone-marrow-derived mesenchymal stem cells, are more closely related to dental tissues. This paper gives a short overview of stem cells of dental origin.
|A Novel Multiplexed Digital Gene Expression Technology|
Gary K. Geiss1,#, Roger Bumgarner2, Brian Birditt1, Timothy Dahl1, Naeem Dowidar1, Dwayne L. Dunaway1, H. Perry Fell1, Sean Ferree1, Renee D. George1, Tammy Grogan1, Jeffrey J. James1, Malini Maysuria1, Jeffrey D. Mitton1, Paola Oliveri4, Jennifer L. Osborn3, Tao Peng2, Amber L. Ratcliffe1, Philippa J. Webster1, Eric H. Davidson4, and Leroy Hood5
We describe a novel platform, the nCounter Analysis System, for sensitive, highly multiplexed, digital gene expression analysis based on NanoString’s novel molecular barcoding technology. Detection of individual mRNA molecules using an assigned sequence of six different fluorescent spots per probe are detected, and then the number of times that code sequence appears in a sample are counted.
|The Aetiological, Clinical and Metabolic Profile of Hypokalemic Periodic Paralysis in an Indian Population|
Jubbin Jacob, Narsing Rao, Mathew John, Simon Rajaratnam , Nihal Thomas , M.S.Seshadri
Hypokalemic periodic paralysis (HPP) represents a heterogeneous group of disorders that presents with acute muscular weakness. Familial periodic paralysis is the commonest cause of hypokalemic periodic paralysis in Caucasians and thyrotoxic paralysis in the Orientals. In this analysis, we have complied the etiological factors that appear to be more dominant in Indian population
|qPCRas a primary screen in drug discovery|
GauravJaggi, Frank Boeckler, Andreas Joerger and Alan Fersht
We report the use of qPCR technique to follow the thermal unfolding of proteins by the binding of the dye SYPRO Orange, and exploit its potential as a robust and high-throughput primary screen for small molecule drug discovery.
|Targeting Inflammatory Cytokines Using Adenoviruses: gene delivery of biological therapies in ovarian cancer|
Michael A. Salako, Hagen Kulbe, Iain A. McNeish, Frances R. Balkwill
Constitutive TNF-alpha expression is characteristic of the malignant ovarian surface epithelium. Adenoviral mutants hold great promise as gene therapy vectors but their efficacy is hindered by an inflammatory cascade orchestrated by TNF-alpha. We found that delivering TNF-alpha shRNA to ovarian cancer cells using oncolytic adenoviruses could reduce the inflammatory cascade generated by adenoviruses and also had direct anti-tumour activity on the cancer cells.