For this reason, only changes in blood flow that persist for some time can be recorded. In the present experiments (study 1), the transverse colon was instrumented with a needle-type platinum EPZ-5676 clinical trial electrode, while a reference electrode was placed in the peritoneal cavity. As has been reported for the stomach (Holzer et al., 1991), the platinum electrode was inserted from the serosa tangentially to the colon at an angle of about 30 degrees and positioned in the submucosa of the colon. The reproducibility of placing the needle electrode in the submucosal layer of the colonic wall was confirmed by histology. To this end, the needle electrode was stained with blue ink before being placed in the colonic wall. After removal of the electrode, the tissue was rapidly excised and frozen.

Sections (35 ��m thick) of the tissue perpendicular to the needle track were taken in a cryostat and examined under a microscope. The needle track was invariably localised to the submucosal layer of the colonic wall, as observed in three rats. The measurement of CBF via the clearance of inhaled hydrogen gas was discontinuous, as the experimental protocol involved alternating 15 min periods of saturation, and desaturation, of the tissue with hydrogen gas (Holzer et al., 1991; Heinemann et al., 1999). The current representing the actual hydrogen concentration at the site of the electrode was taken up by a polarographic unit, amplified, digitised at a frequency of 1 Hz and recorded on a personal computer with a custom-made software (Heinemann et al., 1999).

The washout curve was then fitted to a monoexponential curve, the power of which was used to calculate the average CBF during the 15 min period of desaturation (Livingston et al., 1989). Average values of MAP and HR were determined for the same time periods. In addition, the colonic vascular conductance (CVC) value was calculated as CBF divided by MAP. Measurement of CBF with laser Doppler flowmetry The principle of laser Doppler flowmetry is based on the reflectance of monochromatic light by moving particles in the tissue. As the volume of tissue that is sampled is not precisely known, the technique cannot record blood flow in absolute values but in arbitrary perfusion units (PUs). The uncertainty about the tissue volume sampled also makes it difficult to compare recordings taken from different animals with each other because the vascular geometry of the tissue sample (i.

e. the relative proportion of arteries, arterioles and capillaries) may differ substantially but cannot be controlled adequately. The major advantages of the Carfilzomib technique are that it provides continuous recordings of blood flow and that it is able to detect rapidly occurring and short-lasting changes in this variable following an intervention. In all studies, except study 1, CBF was measured by laser Doppler flowmetry.