Microsoft word - program-3d-o2-no-course-aug2011.doc

Preliminary program
1. 9 a.m. Lecture:
“New Updates in Detection and Imaging of ROS in vitro and in vivo using EPR”
(Fink B., M.D. Noxygen Science Transfer & Diagnostics GmbH)


2. 10:00 a.m. (15 min coffee break)

Introduction in use of cyclic hydroxylamines for detection of O ●
a. Preparation of Krebs-HEPES buffer, spin probes, and other solutions b. Demonstration of preparation of aorta sections/heart tissue c. Time dependence of O ● 2 - production in isolated aorta and heart tissue, d. Preparation of heart tissue samples for detection in liquid nitrogen (frozen) 3. 1 p.m. Lunch break

4. 2 p.m.
a. Oxygen dependence of ROS- production in rat aorta/heart tissue/blood cells
b. Temperature dependence of ROS- production in cell suspension (peripheral blood / mononuclear
cells/cultured cells) (demonstration of “Temperature Controller”)
c. Problems, pitfalls, advantages of ROS- detection using cyclic hydroxylamines and EPR
5. 4 p.m. (15 min coffee brake)
2 (basal and stimulated O2 - production, PEG-SOD-inhibition) 2 (basal and stimulated O2 - production, SOD-inhibition) c. Effects of hypertonic and hypotonic conditions on ROS production in suspension of cells;
6. 6 p.m. (15 min coffee break)
Acquisition and Evaluation software (Win-Acquisition, Win-EPR)
a. Processing of EPR spectra
b. Calibration of EPR system/ Quantification of free radicals production
a. Introduction in analysis of oxygen tension and oxygen consumption in suspension of cells (peripheral blood / mononuclear cells/ cultured cells (demonstration of Gas Controller))
b. Simultaneous detection of oxygen consumption and ROS formation in suspension of cells (peripheral blood / mononuclear cells/ cultured cells) – NEW! c. Signal calibration, problems, pitfalls, advantages of pO2/ROS - detection in vitro and in vivo 2. 11 a.m. (15 min coffee break)
a. Introduction in use of cyclic hydroxylamines for analysis of O ● b. Oxygen dependence in production of O ●
(demonstration of “Gas Treatment Chamber)
2 production in cells/tissue/mitochondria 3. 1 p.m. Lunch break
a. Vitality/ Condition Test - analysis of ROS production in human blood – NEW! b. Inflammatory Response Assay - analysis of TNF-alpha induced ROS formation c. Problems, pitfalls, advantages of ROS - detection in vitro and in vivo 5. 4 p.m. (15 min coffee break)
a. Real time detection of ROS in vivo: NEW!
i. Effect of vasoactive drugs
ii. Environmental influence
- heat shock, exposure to cigarette smock iii. Effect of food ingredients
- vitamins, herbal extracts, glucose overdose iv. Inflammatory response

In case of request
b. Real time detection of ROS and oxygen consumption in zebrafish embryos: NEW! - perfluorooctane sulfonate, TiO2 nanoparticle 6. 6 p.m. Discussion (coffee break)
1. 9 a.m. Lecture: “Endothelial Control of Vasomotion and Nitric Oxide Production. Analysis
of NO Using Colloid Fe(DETC)2 and EPR”
(Fink B., M.D. Noxygen Science Transfer & Diagnostics GmbH, Germany)

2. 10 a.m.
a. Introduction in use of colloid Fe(DETC) for detection NO-production in cultured cells and b. Detection of NO in cultured cells and isolated aorta, (basal and stimulated NO production / L-NAME inhibition) c. Simultaneous detection of NO and ROS under simulated in vivo conditions (shear stress, pressure, oxygen concentration, temperature) in cultured cells (demonstration of Shear Stress Controller (laminar, pulsatile, oscillatory shear stress) - NEW! 2. 12 a.m. (15 min coffee break)
a. EPR settings for detection of NO (sweep, sweep time, gain, modulation amplitude, time b. Calibration of EPR-signal for calculation of NO-production c. Problems, pitfalls, advantages of NO detection using colloid Fe(DETC)2 and EPR
3. 1 p.m. Lunch break

4. 2 p.m.
a. Simultaneous detection of ROS and NO in vivo (mouse/rat) b. Detection of NO using new spin label in cultured cells – NEW! c. Problems, pitfalls, advantages of simultaneous NO/ROS detection in vitro and in vivo
5. 4 p.m. (10 min coffee break)
a. Detection of circulating NO (NOHb concentration) in rodent and human blood
b. Calibration of EPR-signal for calculation of NO-production
c. EPR settings for detection of NO production in fluid and frozen samples (sweep, sweep time,
gain, modulation amplitude, time constant) d. Problems, pitfalls, advantages of NO detection in vitro and in vivo
6. 6 p.m. (15 min coffee break)

7. Closing remarks, discussion

Source: http://www.noxygen.de/pub/Program-3d-O2-NO-Seminar-Aug2011.pdf