Effect on protein carbonylation, ROS production, lipid peroxidation and co-enzyme Q9 levels in the absence (Pb-acetate) and existence of AEEF (Pb-acetate +AEEF) in experimental mice.

<p>Results were represented as mean ± SE (n = 6). ^$ Results differed significantly (p < 0.05) from Pb-acetate control. ^#Results differed (p < 0.01) significantly from normal control. * Results significantly (p < 0.05) differed from Pb-acetate control. ** Results differed (p < 0.01) significantly from Pb-acetate control.</p

Effect on cellular redox systems in the absence (Pb-acetate) and existence of AEEF (Pb-acetate +AEEF) in experimental mice.

<p>Results were represented as mean ± SE (n = 6). ^$ Results differed significantly (p < 0.05) from Pb-acetate control. ^#Results differed significantly (p < 0.01) from normal control. * Results significantly (p < 0.05) differed from Pb-acetate control. ** Results significantly (p < 0.01) differed from Pb-acetate control. CAT unit “U” is defined as H₂O₂ consumed/minute while SOD unit, “U” is defined as inhibition (μ-moles) of NBT-reduction/min.</p

A Schematic overview of <i>in vivo</i> experimental protocol.

<p>A Schematic overview of <i>in vivo</i> experimental protocol.</p

Histological assessments of testes along with histo-quantification data of experimental mice in the absence (Pb-acetate) and existence of AEEF (Pb-acetate + AEEF).

<p>Histological sections 100 x (Panel A) and 400 x (Panel B) of testes. The section of testes of normal control mice exhibited all stages of spermatogenesis, while, testes of Pb-acetate treated mice showed disruption of normal arrangement of seminiferous tubules also in the process of spermagenesis. Yellow arrows represented spermatogonia close to the basement membrane; red arrow represented primary spermatocytes; green arrows represented round spermatids; blue arrows denote elongated spermatids; black arrows represented complete spermatozoa. However, AEEF treatment could attenuate the Pb-acetate mediated toxic manifestations in testes of mice. Panel C. The Johnsen score was measured (400 X, comprising one seminiferous tubule). Seminiferous tubule at Johnsen score 10 presenting all stages of spermatogenesis. The Jhonsen score is descending with the toxic occurrence within testicular tissues. Results were expressed as mean ± SE, (n = 60). ^# Results differed significantly (p < 0.01) from normal control. ^**Results significantly (p < 0.01) differed from Pb-acetate control.</p

Effect on Pb accumulation (Panel A), DNA fragmentation (Panel B) DNA oxidation (Panel C) and ATP levels (Panel D) in the absence (Pb-acetate) and existence of AEEF (Pb-acetate +AEEF) in heart, kidney, liver, brain and testes in mice.

<p>Results were denoted as mean ± SE (n = 6). ^#Results differed significantly (p < 0.01) from normal control. * Results significantly (p < 0.05) differed from Pb-acetate control. ** Results significantly (p < 0.01) differed from Pb-acetate control.</p

Schematic presentation of the hypothesis developed in this study regarding the overall protective mechanism of AEEF against Pb toxicity.

<p>The dark blue arrows indicate the cellular events involved in Pb-induced pathophysiologies. The green lines denoted the activity promoted (+) by AEEF, while, red lines denoted the activity restricted (-) by AEEF.</p

The effect on cellular ROS production, degree of lipid peroxidation, protein carbonylation, endogenous redox systems in the absence (Pb-acetate) and presence of AEEF (Pb-acetate + AEEF) in isolated mouse hepatocytes.

<p>Panel A. Effect on ROS-generation in Pb-exposed hepatocytes was measured by fluorescence microscopy in the absence (Pb-acetate) and presence of AEEF (Pb-acetate + AEEF). Panel B. Effect on protein carbonylation, lipid peroxidation and endogenous redox status in the absence (Pb-acetate) and existence of AEEF (Pb-acetate + AEEF). Results were represented as mean ± SE (n = 3). ^#Results were significantly (p < 0.01) different from normal control. *Results were significantly (p < 0.05) different from Pb-acetate control. **Results varied significantly (p < 0.01) from Pb-acetate control. SOD unit, “U” was defined as inhibition (μ-moles) of NBT-reduction/min while CAT unit “U” was defined as H₂O₂ consumed/minute.</p

Histological assessments of livers along with histo-quantification data of experimental mice in the absence (Pb-acetate) and existence of AEEF (Pb-acetate + AEEF).

<p>Histological sections 100 x (Panel A) and 400 x (Panel B) of livers. The liver section of normal control mice showed normal portal vein and hepatocytes. Pb-acetate treated liver section exhibited dilated portal vein (green arrow), fatty degeneration (blue arrows), vacuolated cytoplasm (red arrows), apoptosis (yellow arrows) and leucocytes infiltration (black arrows) when compared with the section of normal control liver. Panel C. The dilation of portal vein is denoted as % of the blank area comparative to the whole area of the photomicrograph (400 x, arbitrarilynominated areas comprising one portal vein were selected). Panel D. The incidence of inflammation was presented as the % of the inflamed hepatocytes region comparative to the whole area of the photomicrograph (100 x, arbitrarily selected area in portal vein were designated). Values were expressed as mean ± SE, (n = 60). ^# Results significantly (p < 0.01)differed from normal control. ^**Results significantly (p < 0.01) differed from Pb-acetate control.</p

Immunoblot analysis of intrinsic factors of apoptotic event in the absence (Pb-acetate) and presence of AEEF (Pb-acetate + AEEF) in isolated murine hepatocytes.

<p>The comparative band intensities were evaluated and the normal control band was assigned a random value of 1 [The ratio of the band intensity of signal protein under normal control and the band intensity of β actin under normal control group was unified and subsequently the band intensities of other groups were obtained by multiplying with same factor used during unification of the protein expression in normal control group]. Loading protein used was β actin. Results were expressed as mean ± SE (n = 3). ^#Values significantly (p < 0.01) differed from normal control. *Results significantly (p < 0.05) differed from Pb-acetate. ** Results significantly (p < 0.01) differed from Pb-acetate control.</p

The effect on cell viability assay in the absence (Pb-acetate) and presence of AEEF (Pb-acetate + AEEF) in isolated murine hepatocytes.

<p>Panel A. Effect on cell viability assayed in the absence (Pb-acetate) and presence of AEEF (Pb-acetate + AEEF). Panel B. Hoechst staining of mouse hepatocytes in the absence (Pb-acetate) and presence of AEEF (Pb-acetate + AEEF). The white arrows represented the heterogeneous intensities and chromatin condensation of nuclei. Results were represented as mean ± SE (n = 3).</p