ischaemic preconditioning of the human heart. - Leicester Research ...

except some vacuoles were present adjacent to the mitochondria, and grade 3
(severe damage): reduced staining of cytoplasmal organelles, clumped chromatin
material, wavy myofibrils and granularity of cytoplasm or cells with contraction
band necrosis.
F. Preparation of neutrophils
Heparinized, venous blood (4 ml) was obtained from patients the day before surgery
in accordance with a protocol approved by our local ethics committee.
Poly morphoneutrophi Is (PMNs) were isolated as described previously using
Hypaque-Fico density-gradient dextran sedimentation [821. Purified PMNs were
resuspended in PBS and kept on ice until use. PMNs (1-10 x 10' cells/ml) were
added to the slices during the experimental protocol.
2.7 Statistical Analysis
Data was entered into SPSS 9.0 spreadsheet (Chicago, IL, USA). Data were
expressed as means ± SEM. One way analysis of variance (ANOVA) was used for
the comparisons of more than two means. ANOVA for repeated measurements
was
used to test the significance of mean values overtime. Ap value less than 0.05 was
considered to be statistically significant.
2.8 RESULTS:
A. Stability of the preparation (Study 1)
LDH leakage: The leakage of LDH into the medium during the incubation period, as
shown in Figure 2.5, represents the accumulation of the enzyme overtime. LDH
leakage steadily increased during the first 12 hr of incubation, so that it rose trom
2.8±0.2 U/g wet wt at 0.5 hr to 10.4-+0.7 U/g wet wt at 12 hr. However, if the
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