Acetonic and Methanolic Extracts of Heterotheca inuloides

The present study was designed to test the hypothesis that the acetonic and methanolic extracts of H. inuloides prevent carbon tetrachloride-(CCl4) induced oxidative stress in vital tissues. Pretreatment with both H. inuloides extracts or quercetin attenuated the increase in serum activity of alkaline phosphatase (ALP), total bilirubin (BB), creatinine (CRE), and creatine kinase (CK), and impeded the decrease of γ-globulin (γ-GLOB) and albumin (ALB) observed in CCl4-induced tissue injury. The protective effect was confirmed by histological analysis with hematoxylin-eosin and periodic acid/Schiff's reagent. Level of lipid peroxidation was higher in the organs of rats exposed to CCl4 than in those of the animals treated with Heterohteca extracts or quercetin, and these showed levels similar to the untreated group. Pretreatment of animals with either of the extracts or quercetin also prevented the increase of 4-hydroxynonenal and 3-nitrotyrosine. Pretreatment with the plant extracts or quercetin attenuated CCl4 toxic effects on the activity of several antioxidant enzymes. The present results strongly suggest that the chemopreventive effect of the extracts used and quercetin, against CCl4 toxicity, is associated with their antioxidant properties and corroborated previous results obtained in liver tissue

Gas Chromatography-Mass Spectrometry Analysis of Ulva fasciata

The chemical composition and biological properties of Ulva fasciata aqueous-ethanolic extract were examined. Five components were identified in one fraction prepared from the extract by gas chromatography-mass spectrometry, and palmitic acid and its ethyl ester accounted for 76% of the total identified components. Furthermore, we assessed the extract’s antioxidant properties by using the DPPH, ABTS, and lipid peroxidation assays and found that the extract had a moderate scavenging effect. In an experiment involving preexposition and coexposition of the extract (1–500 µg/mL) and benzo[a]pyrene (BP), the extract was found to be nontoxic to C9 cells in culture and to inhibit the cytotoxicity induced by BP. As BP is biotransformed by CYP1A and CYP2B subfamilies, we explored the possible interaction of the extract with these enzymes. The extract (25–50 µg/mL) inhibited CYP1A1 activity in rat liver microsomes. Analysis of the inhibition kinetics revealed a mixed-type inhibitory effect on CYP1A1 supersome. The effects of the extract on BP-induced DNA damage and hepatic CYP activity in mice were also investigated. Micronuclei induction by BP and liver CYP1A1/2 activities significantly decreased in animals treated with the extract. The results suggest that Ulva fasciata aqueous-ethanolic extract inhibits BP bioactivation and it may be a potential chemopreventive agent

Associations of the clinicopathological factors with CYP2E1 and CYP2W1 mRNA expression.

§<p>Mann-Whitney rank test.</p>£<p>Spearman rank correlation test.</p><p>*Statistically significant p<0.05.</p

Levels of normalized mRNA Expression of (a) CYP1B1, (b) CYP2E1, (c) CYP2W1, (d) CYP3A4 and (e) CYP3A5 in 13 pairs of tumor and corresponding normal adjacent tissue from patients with RMS.

<p>Transcript levels of each CYP gene were normalized to β-actin and expressed per μg of total RNA. Values represent means and average of standard deviations of expression levels determined in triplicate. *p<0.05.</p

Demographic and clinical features of patients involved in the study.

<p>*Primary sites: EX = Extremities; GU = Genitourinary; H&N = Head and Neck.</p

Expression levels of CYP genes displayed as cycle threshold (Ct) values in 13 pairs of tumor and corresponding normal adjacent tissue [Mean Ct values (mean β-actin Ct value)].

<p>BLQ: Below limit of quantification (Ct≤38); High mRNA levels (Ct ≤35) are in bold; *Mean ± standard deviation of samples with Ct ≤38 of each CYP gene; n = number of samples with Ct value ≤38. Experiments performed in triplicate.</p

Western blots analysis of CYP proteins.

<p>(A) Representative immunoblots of CYP1B1, CYP2E1, CYP2W1, CYP3A4, and CYP3A5 proteins from non-tumor tissues (N) and tumor tissues (T) of RMS patients: 2, 5, 6, and 12. (B) The levels of β-actin were analyzed to ensure samples’ loading amount.</p

Summary of RT qPCR and Western data in four different patients.

<p>N, normal adjacent tissue; T, tumor tissue. For RT qPCR (PCR) data: −, undetectable amounts of mRNA; +, detectable mRNA expression; +^#, high amounts of mRNA. For Western (W) analysis: −, not detected; +, positive detection; (+), weak detection.</p