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192 Abstracts / Toxicology L

eurotoxicity (DNT). There is concern that exposures to envi-onmental chemicals contribute to the increasing incidence ofeurodevelopmental disorders in children. However, due to lackf DNT studies only very few substances have been identified asevelopmental neurotoxicants. This study aimed to develop an

n vitro approach using metabolomics and transcriptomics for DNTssessment. A 3D rat primary neuronal organotypic model wasxposed to well known or suspected (developmental) neurotoxi-ants including pesticides (carbaryl, chlorpyrifos, lindane maneb),rugs (phenytoin, valproic acid) and metals (cadmium, lead) fromay 7 up to 21. Quantitative measurement of genes expressed inifferent cell types (nestin in neural precursor cells, neurofilament-00 in neurons, S100� in astrocytes and myelin basic protein

n oligodendrocytes) and mass spectrometry based metabolomicseasurements for some compounds were performed. Treatmentith the different compounds significantly modified the expression

f the selected genes. Moreover, the mass spectrometry analy-is showed differences in metabolite levels between control andreated cells. Further analysis of the altered metabolites should give

echanistic insight into the DNT of these compounds. This studyemonstrates that gene expression and metabolomic analysis cane sensitive endpoints for DNT assessment. Funded by the Swedishesearch Council and the FDA.

oi:10.1016/j.toxlet.2012.03.689

31-10rotective effects of Teucrium polium L. on acetaminophen

nduced hepatotoxicity

eibatullah Kalantari, Ebrahim Azemi, Hossien Frouzandeh, Iranashidi

Ahvaz Jundishapur University of Medical, Iran

Purpose: Acetaminophen is widely used as an over-the-ounter analgesic and antipyretic drug. Intake of a large dose ofcetaminophen may result in severe hepatic necrosis. In this studyhe protective effect of Teucrium polium L. (Lamiaceae) extract oncetaminophen induced hepatotoxicity in mice was carried out.ethods: From the dried powder of Teucrium polium L. hydroal-

holic extract was prepared. Animals were divided into six groups,ach group consist of 8 mice. Group one received normal salinenegative control), group two, received only crude extract of T.olium L. (500 mg/kg) for its toxicity evaluation and group threeeceived acetaminophen in dose of 500 mg/kg (positive control).roups four, five and six were received crude extract orally inoses of 125, 250, 500 mg/kg, respectively, for five days then 1 hfter the last administration of the crude extract acetaminophenas given. Then on the day 6th, animals were sacrificed, blood wasithdrawn to determine serum enzymes activities of ALT, AST andLP also to measure serum levels of directed and total bilirubin.iver was removed for histological examination. Results and conclu-ion: The results of this study showed that Teucrium polium L. hadiver protective effect in all doses applied, but the most significantrotection was observed in doses of 250 and 500 mg/kg (P < 0.05).

lso these findings were supported and confirmed by histologicalxamination.

oi:10.1016/j.toxlet.2012.03.690

211S (2012) S43–S216

P31-11Intrathecal stem cells in rotenone induced Parkinson’s disease(PD) model in mice

Mohamed Salama 1, Dina Tantawy 1, Sahar Dakroury 1, MahmoudAl-Husseiny 1, Sara El-Dousoky 1, Hassan Abdelghaffar 1,Abdelaziz Ghanem 1, Oscar Arias-Carrion 2, Seham Gad El-Hak 1

1 Mansoura University, Egypt, 2 Munich Technical Institute, Germany

The aim of the present study was to evaluate the neuroprotectiveeffects of Wharton Jelly Cells (WJCs) in a progressive model of PD inmice induced by rotenone. In our study the mice were exposed todaily doses rotenone diluted in CMC 0.5% (30 mg/kg) administeredorally through gavage for 28 days. The mice were divided into 3groups; the first group (control) received CMC 0.5% only; the sec-ond group received the daily doses of rotenone; the third groupreceived the rotenone doses, however, they received intrathecalWJCs in the 14th day of the experiment. The mice were evaluatedregularly (weekly) for locomotor disturbances through behavioraltests (vertical and horizontal grid tests). Following scarifice, thebrains were divided into 2 halves, the first for immunohistochem-ical evaluation using anti-TH antibodies, followed by stereologybased counting of dopaminergic neurons in substantia nigra (SNpc),and image analysis (imageJ system) of fibers density in corpus stria-tum. The other half of the brain was evaluated regarding dopamineand its metabolites level using HPLC with electrochemical detector.The rotenone model was established based on behavioral tests thatrevealed significant impairement in locomotor functions comparedto the control group. Postmortem assessment revealed significantdegeneration of SNpc neurons and striatal fibers, moreover chemi-cal measurement revealed significant decrease in dopamine levels.The third group that received intrathecal WJCs revealed significantimprovement in behavioral tests. Moreover, histopathological eval-uation of SNpc and striata revealed significant neuro-regenerationcoupled with significant elevation of dopamine levels compared torotenone group.

doi:10.1016/j.toxlet.2012.03.691

P31-12Evaluation of lineage proliferation markers in rat bonemarrow by flow cytometry

Minna Suomela, Alaa Saad

AstraZeneca R&D Innovative Medicine, Sweden

In addition to histopathology, bone marrow analysis by flowcytometry has been utilized in toxicity studies for the evaluationof cellularity and cellular composition. The available methods donot provide information on the proliferating and mature/maturingcompartment of various cell lineages. The addition of this informa-tion would improve the assessment of various toxic alterations inthe bone marrow. The aim of this study was to evaluate the addi-tion of lineage proliferation markers to the existing method. DNAProbe (DRAQ5TM) was used in combination with CD45:FITC (leuko-cyte common antigen) and CD71:PE (transferrin receptor). The keyfeatures of DRAQ5TM is its stoichiometric binding to DNA (permitsDNA content analysis), membrane permeant dye (omit the needof permeabilisation step) and its emission spectrum allows the

combination with other fluorescence probes (i.e. FITC and PE). Themorphological complexity of cells combined with antigen expres-sion was used to gate the main lineages (erythroid, myeloid andlymphoid) and a histogram of cellular DNA content was plotted