Michelle Hermiston, MD, Ph.D.
Assistant Professor, Department of Pediatrics University of California San Francisco, USA
Interrogating Signaling Networks at the Single Cell Level In Primary Human Patient Samples
Multiparameter phosphoflow cytometry is a highly sensitive proteomic approach that enables monitoring of biochemical perturbations at the single cell level. By combining antisera to cell surface markers and key intracellular proteins, perturbations in signaling networks, cell survival and apoptosis mediators, cell cycle regulators, and/or modulators of other cellular processes can be analyzed in a highly reproducible and sensitive manner in the basal state and in response to stimulation or drug treatment. Advantages of this approach include the ability to identify the biochemical consequences of genetic and/or epigenetic changes in small numbers of cells, to map potential interplay between various signaling networks simultaneously in a single cell, and to interrogate potential mechanisms of drug resistance or response in a primary patient sample. Application of this technology to patients with acute lymphoblastic leukemia or the autoimmune disease systemic lupus erythematosus (SLE) will be discussed.
Bodo Eickhoff, Ph.D.
Senior Vice-President, Head of Sales and Marketing for Roche Applied Science, Germany
New paths for treatment of complex diseases: target combinatorial drug therapy
Several types of diseases show a complex pathogenesis and require targeted as well as combinatorial drug treatment. A classical example, Tuberculosis, was thought for decades to be managable by triple therapy, however now requiring new therapeutic approaches due to multi drug resistant strains. HIV and AIDS can only be kept under control by combinations of specific, virus-protein targeted drugs, requiring constant monitoring of resistance patterns and modulation of drug combinations during life-long therapy. As a third example, Cancer in all its different variations, requires detailled molecular understanding to enable targeted therapy. New technologies provide more and in depths molecular insights into pathomechanisms and resulting treatment options. However, is there an alternative way to approach complex diseases by holistic models? Can restoring of apoptosis-capabilities of transformed cells be an example of such an alternative path? How do we in future adress major unresolved topics like increasing drug resistance in bacterial infections, lack of anti-viral drugs, treatment of parasite diseases like Malaria, and newly emerging infectious diseases in research and fast translation of these results into diagnosis and treatment?
Syed Salman Lateef and Vinayak A K
Development of Supercritical Fluid Chromatography methods for the replacement of existing USP Normal phase liquid chromatography methods
Normal phase liquid chromatography methods often have long run times and involve environmentally toxic/costly solvents. Supercritical chromatography methods on the other hand are faster, inexpensive, and eco-friendly. The low viscous supercritical carbon dioxide operates at high flow rates compared to LC without losing separation efficiency. In this work, SFC methods are developed to replace three United States Pharmacopeial (USP) normal phase achiral methods – prednisolone, tolazamide and cholecalciferol. System suitability parameters of the normal phase method are compared against the SFC method. Precision, linearity and robustness of the new SFC methods are demonstrated. SFC methods were found to be cost effective in terms of analysis time and solvent savings. The SFC method does not require purchase and disposal of expensive environmentally hazardous chemicals. Hence, the newly developed SFC method provides a faster and safer solution.