Generation and Action of Reactive Species 1 Trichloroethylene’s Dermal Toxicity Involves Activation of PKC and Intracellular Calcium Ions Burst: Prevention by Natural Flavonoid Naringenin Through Antiapoptotic Strategy Farrah Ali1 and Sarwat Sultana1 1Jamia Hamdard, India Trichloroethylene is a widespread industrial solvent responsible for severe liver dysfunction and dermatitis in occupationally exposed humans. Dermal toxicity is the major exposure related concern. There are several previous studies which implicate the hypersensitivity reactions due to TCE but there is no substantial research delineating the mechanism underlying the TCE induced dermal toxicity. Here we present the data which eases our understanding towards TCE toxicity and we are also able to show the interception of these toxic manifestations through dietary flavanone Naringenin. In this study we have used in vitro system of HaCaT cells which has provided clear results without the complex cell and extracellular matrix interactions and uncontrolled hormonal effects which are characteristic of in vivo system. We observed that there was marked decrease in the cell viability dose dependently after 24 hours exposure and the mode followed by cellular death was apoptosis. There was significant depletion in major antioxidative enzyme activity and subsequent increase in intracellular ROS generation and Nitric Oxide production. Subsequently ROS generation lead to mitochondrial membrane depolarization and cell cycle dysfunction. The key finding was significant increase in intracellular Calcium ions that could be in conjunction with the activation of Protein kinase C. Furthermore, comet assay data inferences significant DNA damage thus cueing towards genotoxic effect of TCE leading to apoptosis as confirmed by Annexin V-Cy3 fluorescent microscopy. We also observed the protein expressions of major inflammation dependant molecular markers NFkB and COX-2 expression and observed that there was significant up regulation in their expressions induced due to TCE. Naringenin, dietary flavanone found abundantly in grapefruit was observed to be typically successful strategy against TCE induced dermal toxicity and genotoxic effects.

2 Effect of the Medium on the Rate of Oxidative Attack Promoted by Photo-Excited TMPyP and TPPS4 Juliana C. Araujo-Chaves1, Cintia Kawai1, Sandra M.S. Pinto1, Fernanda Bettanin1, Camila O.C.C. Sanches1, Nasser Daghastanli2, and Iseli L. Nantes1 1UFABC - CCNH, 2UFABC – CECS, Brazil Porphyrins are largely used as photosensitizers in photodynamic therapy. In the present study, the rate of oxidative damage promoted by photosensitized, the cationic TMPyP (meso-tetrakis (N-methyl-4-pyridyl) porphyrin) and the anionic TPPS4 (meso-tetrakis(4-sulfonatophenyl)porphyrin), was measured by meas-uring the fluorescence increase correlated to the oxidation of H2DCFDA. The fluorescence resulting from the oxidation of H2DCFDA by the porphyrins was monitored in aqueous glucose

solution and in the presence of DPPC/TOCL (dipalmitoyl-sn-glycero-3-phosphocholine / tetraoleoyl cardiolipin). The irradiation of the samples with white LED source (emission from 450 to 650 nm) promoted an increase of H2DCFDA fluorescence at the following rates: TPPS4 = 1.8 x 10-3.s-1 and TMPyP = 4.5 x 10-3 s-1 in aqueous homogeneous medium and TPPS4 = 1.2 x 10-3 s-1 and TMPyP = 2.6 x 10-3.s-1 in the negatively charged DPPC/TOCL membranes. In both investigated media the photodynamic action of TPPS4 was not accompanied porphyrin bleaching while bleaching of TMPyP was observed only in homogeneous medium. In the presence of DPPC/TOC, TMPyP presented blue shift of Q bands. The results are consistent with the differentiated affinities of the porphyrins to the model membrane, since cationic TMPyP binds to anionic DPPC/TOCL and TPPS4 remains in bulk water even in the presence of liposomes. FAPESP, CNPq and UFABC

3 Reactive Oxygen Species Interactions with Plasmodium Parasites During Uncomplicated Malaria Infection in Ghanaian Children Asmah Harry Asmah1, Sarsah Isaac1, Listowel Asare1, Selorme Adupko2, Ben Gyan2, Micheal Ofori2, Edwin Wiredu1, Patrick Ayeh-Kumi1, and Charles Brown1 1University of Ghana School of Allied Health Sciences, 2Noguchi Memorial Institute for Medical Research, Ghana During malarial infection oxidative stress increases as more reactive oxygen species (ROS) are generated. This induces tissue damage and also kills Plasmodium parasites. To circumvent biological injury caused by the oxidative stress, the anti-oxidant enzyme superoxide dismutase (SOD) converts these unstable free radicals into H2O2, which are removed by catalase and glutathione systems. Unlike the host, the parasite is not efficient in circumventing the oxidative stress and to understand this we evaluated the relationship between ROS levels and parasitaemia during uncomplicated malaria infection. Blood samples were collected from 50 malaria infected children and 49 non-infected controls between 0.5 and 12 years in Sogakope, Ghana. Malaria diagnosis and parasite density were determined by light microscopy while haemoglobin (Hb) levels, RBC, platelet (PLT), total WBC and neutrophils counts were measured with the Mindray BC – 3000 Plus auto-haematological analyser. Blood SOD activity to quantify the amount of ROS generated was measured using SOD Assay Kit-WST. Parasite DNA damage was evaluated by Comet Assay using a commercial kit (TrevigenTM). Mean neutrophils, RBC, PLT count, Hb level and SOD% inhibition rate for malaria cases were 6.3 x 109/L, 4.4fl, 165.9/L, 10.4 g/dL ± 1.5, and 14.0% respectively; that of controls were 7.5 x 109/L, 4.9 fl, 262.7/L, 11.0 g/dL, and 17.0% respectively. Inter – group comparison of means of Hb, RBC and PLT showed significant differences (p 0.05). Comet assay gave a clear picture of parasites’ DNA damage. There was positive correlation between SOD and neutrophils, and WBC with parasitaemia levels in study subjects. In conclusion, the study demonstrated ROS-mediated damage of malaria parasite DNA in infected RBCs and showed that parasitaemia levels influences SOD inhibition rate.

SFRBM 2012 S15

doi:10.1016/j.freeradbiomed.2012.10.028

doi:10.1016/j.freeradbiomed.2012.10.029

doi:10.1016/j.freeradbiomed.2012.10.030