Assessment of Malondialdehyde (MDA): The level of MDA in brain was found by mixing brain homogenate
(100 ul) with sodium dodecyl sulfate (200 ul), acetic acid (1.5 ml) and thiobarbituric acid (1.5 ml).
extent of Lipid Peroxidation (LPO) and Nitric Oxide (NO) level were assessed in four different tissues homogenate
i.e Liver, Kidney, Pancreas, Muscle.
Catalase activity (figure 4) did not exhibit any statistically significant difference between the liver and lung homogenates
. In the brain homogenates
, the activity significantly decreased in the GR (P<0.001) and DR (P=0.006) groups.
extraction was compared with ultrasonic extraction for crocins from saffron.
were mixed with loading buffer (glycerol 30%, SDS 10%, Tris 0.5 M, bromophenol blue 0.01%, and [beta]-mercaptoethanol) and boiled for 10 min in a water bath.
Based on the comparison of peak areas with standard sample, the concentration of sialic acid in EBN homogenate
was quantified to be 0.45 [+ or -] 0.05g/100mL.
For neutralizing antibody assay, THP-1 cell-derived macrophages were treated with 100 ng/ml LPS for 24 h and then cultured with 2 [micro]g/ml IL-6 neutralizing antibody or IgG isotype control antibody for another 24 h, followed by incubation with 100 [micro]l/ml eutopic versus ectopic endometrial homogenate
for 72 h, or 10% serum from control subjects versus endometriosis patients for 6 days.
The protein concentrations in the homogenate
, supernatant and the extract of pellet were determined by total protein quantitative assay kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) according to the instructions of the manufacturer.
vCJD brain homogenate
was serially diluted to assess detection sensitivity of the PMCA assay; a positive result was obtained down to a [10.sup.-10] dilution.
solution spiked with standard,
In addition, we monitored the levels of anti-oxidative and stress biomarkers in liver tissue homogenates
obtained from these animals.
Each PMCA experiment included a reference vCJD sample (10% brain homogenate
) as a control for the amplification efficiency.