Effect of strain on the expression of metallothionein-1 in mice.

<p>Expression of Mt-1, a stress gene expressed in the liver, as compared to the control gene, Actb, following amplification with quantitative real time PCR. (Mean ± SEM) Mt-1 data was transformed by raising all values to 0.4. A. At 0 hours, a significant difference between BALB/c mice and C57BL/6 mice was observed. Neither was significantly different from CD1 mice in terms of Mt-1 expression. B. At 4 hours, a significant difference was observed from baseline levels for all strains. At this time, the levels were slightly higher for all three strains than the levels observed at 0 hours. F = 3.06; d.f. = 6,00; *P = 0.0 Means with different superscripts within time points are significantly different.</p

Time effect of cytokine expression.

<p>Concentration of serum cytokines observed from quantitative ELISAs. Due to transformations of all data, graphs are shown with arbitrary units. (Mean ± SEM) A. A significant difference was observed in the expression of MIP-1β, a chemokine that attracts macrophages, over the three time points tested. MIP-1β data was transformed by raising all values to 0.4. B. A significant difference was observed in expression of IL-1β, a proinflammatory cytokine, in the three time points tested. IL-1B, unlike others tested, did not show as high variation over the three time points. IL-1β data was transformed by raising all values to 0.25. C. A significant difference was observed in the expression of TNF-α, a proinflammatory cytokine, over the three time points tested. TNF-α data was transformed by raising all values to 0.5. Means with different superscripts within cytokine bars are significantly different. P < 0.05.</p

Body temperature changes over time following LPS exposure.

<p>Following a non-lethal dosage of 5mg/kg of LPS, mice were tested over a 9-hour time period for changes in body temperature. (Mean ± SEM) These were plotted by sex and strain. A. A significant difference in temperature was observed between male and female BALB/c mice at 3 hours and at 4 hours following LPS injection. B. A significant difference in temperature was observed between male and female CD1 mice at 2 hours, but not at any other time points measured. C. A significant difference was observed between male and female C57BL/6 mice at 4 and 5 hours following LPS injection. *P < 0.05.</p

Effect of sex and strain on the expression of IL-6.

<p>Expression of serum IL-6, a proinflammatory cytokine, tested using quantitative ELISAs. (Mean ± SEM) IL-6 data was transformed by raising all values to 0.3. At 0 hours, a significant difference was observed, with any difference in sex or strain being dependent on the other variable. At 7 hours, a significant difference was observed, with any difference in sex or strain being dependent on the other variable. Differing letters indicate significance. P < 0.05.</p

Beta-fibrinogen expression over time in three strains.

<p>Expression of Fgb mRNA as compared to control Actb mRNA following amplification of both genes with quantitative real time PCR. (Mean ± SEM) Fgb data was transformed by raising all values to 0.2. A. There was a significant strain by time interaction (<i>F</i> = 5.55; d.f. = 6, 91; <i>P</i> < 0.0001), with post-hoc tests showing differences in expression levels between BALB/c and B6 mice at 7 hours, and between BL6 and both other strains at 4 hours B. There was also a sex by time interaction (<i>F</i> = 6.73; d.f. = 3, 91; <i>P</i> = 0.0004)), with higher expression levels in males compared with females at the 2 hour and especially the 7 hour time periods, but the reverse at the 4 hour time period. Means with different superscripts are significantly different. P < 0.05.</p

Effects of ethanol and androgenization on antioxidant defense activity.

<p>A. Hepatic superoxide dismutase (SOD) activity determined by assay kit for control (Ctrl), androgenized (Andro) or androgenized + E2 (Andro + E2) animals maintained on control Lieber-DeCarli (C-LDC) or ethanol-LDC (E-LDC) diet. B. Hepatic catalase activity determined by assay kit for Ctrl, Andro, Andro + E2 rats maintained on C-LDC or E-LDC diets. Activity levels were normalized to total protein. Data are presented as mean values ± SEM. *P<0.05.</p

Effects of ethanol and androgenization on CYP450 expression.

<p>A. Quantitative Real-Time PCR analysis of CYP2E1 mRNA expression in control (Ctrl), androgenized (Andro) and Andro + estrogen (Andro + E2) rats maintained on control Lieber-DeCarli (C-LDC) or ethanol-LDC (E-LDC) diet. B. Representative CYP2E1 protein expression was determined by Western blot analysis (upper panel) and quantified by optical integrated volume (lower panel). Data are presented as mean values ± SEM. Standard square root transformation was performed prior to statistical analysis. *P<0.05. C. Quantitative Real-Time PCR analysis of CYP1A2 mRNA expression compared between groups described in A. Data are presented as mean values ± SEM. *P<0.05. D. Representative CYP1A2 protein expression was determined by Western blot analysis (upper panel) and quantified by optical integrated volume (lower panel). RNA and protein expression were normalized to GAPDH.</p

Effects of ethanol and androgenization on oxidative stress.

<p>A. Representative images of 4-HNE stained formalin-fixed paraffin-embedded liver tissue from rats maintained on control Lieber-DeCarli (C-LDC, top panels) or ethanol-LDC (E-LDC, bottom panels) diet. Panels A and D, control females (Ctrl); B and E, androgenized females (Andro); panels C and F, Andro + estrogen (Andro + E2). PT; portal triad. Caramel brown staining indicating 4-HNE positive cells. B. Hepatic lipid peroxidation was assessed by Thiobarbituric Acid Reactive Substances (TBARS) assay. Liver tissue was homogenized and malondialdehyde (MDA) formation measured. TBARS were normalized to total protein. Data are presented as mean values ± SEM. *P<0.05.</p