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were cut in half at the equator of the bulbus and vitreous and
lens were removed. The ciliary body and iris were carefully
detached from the sclera by cutting the tissue with a cornea
trepan. The tissue samples were pooled and placed in oxyge-
nated modified Krebs solution. After preincubation of 1 h the
multidish wells were cleared of the solution and fresh Krebs
solution and test compounds or solvent were added. The incu-
bation period of 60 min was used. The data measurements have
been described in detail in our recent article [41]. Acetylated
cGMP in the samples was assayed with the [125I]-cyclic GMP
RIA kit (Amersham International, Little Chalfont, Buckin-
ghamshire, UK). Nitric oxide release from the test compounds
was determined spectrophotometrically by measuring nitrate
+ nitrite (NOx) in the incubation medium with the Nitrate/Ni-
trite Colorimetric Assay Kit (Cayman Chemical, Ann Arbor,
MI, USA). The concentrations of test compounds were chosen
on the basis of our previous studies [34]. The differences in the
concentration used are mainly due to the different solubilities
of the compounds in the incubation medium.
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Acknowledgments
This work was financially supported by the Alfred Kordelin
Fund, the Päivikki and Sakari Sohlberg Foundation and the
National Technology Agency, Finland.