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61349201455
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note
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3): δ 7.65 (d, 2H); 7.40 (s, 1H); 7.30-7.15 (m, 7H); 5.60-5.40 (m, 2H); 4.20 (dd, 1H); 3.95 (dd, 1H) 3.70-3.60 (m, 1H); 2.80-2.60 (m, 4H); 2.40-2.20 (m, 4H); 1.95-1.30 (m, 12H).
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18
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61349176844
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note
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High IOP levels were obtained injecting carbomer 0.25% (Siccafluid Farmila THEA Pharmaceutical) 0.1 ml bilaterally into anterior chamber in pre-anesthetized rabbits with 1 ml iv of pentobarbital sodium (Pentothal Abbott, Campoverde di Aprilia, LT). The IOP increase was tested, every 30 min during the first 3 h, and at least three times a day until stabilization. As soon as the IOP was stabilized on high levels, the drugs in study were administered by instillation, once for single dose experiment and every day for 5 days for the repeated dose experiment, and IOP was measured, using a Tono-Pen tonometer (Solan Ophtalmic Products Jacksonville, USA). Aqueous humour (both posterior and anterior chamber fluids) was withdrawn before inducing ocular hypertension, when IOP was increased and after each treatment with the drugs in study and used for GSH and cGMP level determinations.
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20
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61349092877
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note
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Determination of GSH levels in aqueous humour: aqueous humour was centrifuged at 4 °C at 100.000g for 30 min and a fixed volume of supernatant was diluted with phosphate-EDTA buffer to a final volume of 5 ml. For GSH evaluation, 200 μl of 40 mM N-ethylmaleimide was added to 0.5 ml of the supernatant, incubated for 30 min and diluted with 0.1 N NaOH to a final volume of 5 ml. Phosphate-EDTA buffer and 0.1 N NaOH (1.8 and 0.1 ml, respectively) and o-phthaldialdehyde solution (1 mg/ml, 0.1 ml) were added to both mixtures. The samples were incubated at room temperature for 15 min and then the fluorescence evaluated fluorometrically at 420 nm emission and 350 nm excitation. Samples will be calibrated using a standard curve of GSH.
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21
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61349090467
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note
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121I] labelled cGMP (Amersham, Bucks, UK). Five hundred μl of 10% trichloroacetic acid (TCA) were added to the samples which were then centrifuged and TCA extracted with 0.5 M tri-n-octylamine dissolved in 1,1,2-trichloro-1,2,2-trifluoroethane. The samples were then acetylated in acetic anhydrase and the amount of cGMP measured in aqueous phase. The values were expressed as fmoles of cGMP per ml.
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24
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54949084607
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Yang G., Wu L., Jiang B., Yang W., Qi J., Cao K., Meng Q., Mustafa A.K., Mu W., Zhang S., Snyder S.H., and Wang R. Science 322 (2008) 587
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Science
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Yang, G.1
Wu, L.2
Jiang, B.3
Yang, W.4
Qi, J.5
Cao, K.6
Meng, Q.7
Mustafa, A.K.8
Mu, W.9
Zhang, S.10
Snyder, S.H.11
Wang, R.12
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