Abstracts / Journal of Biotechnology 185S (2014) S37–S125 S89

first months of life due to thoracic narrowness and possible heartdefects.

http://dx.doi.org/10.1016/j.jbiotec.2014.07.302

Cytokine gene polymorphism in Wistar AlbinoGlaxo rats from Rijswijk (WAG/Rij) rats whichare genetic models of absence epilepsy

Gönül Gürol 1, Nilgün Tekin 2, Hatice Yildizhan 2,Sibel Sarı 1,∗, Sertan Arkan 3, Nurbay Ates 3, DuyguÖzel Demiralp 2

1 Department of Physiology, Sakarya University,Sakarya, Turkey2 Biotechnology Institute, Ankara University, Ankara,Turkey3 Department of Physiology, Kocaeli University,Kocaeli, Turkey

E-mail address: siibelsari@gmail.com (S. Sarı).

Epilepsy is a neurological disease which affects 1% of the worldpopulation, characterized by recurrent spontaneous seizures dueto hyperexcitability and hypersynchrony of neurons. The role ofgenetic factors has been shown in epileptic syndromes. Humangenome comprises a number of genetic variants as a resultof polymorphisms. In epilepsy, it is possible that these vari-ants cause critical consequences such as reduce or increasethe seizure activity. It is reported that the cytokine gene poly-morphisms are related to increased seizure susceptibility andepileptogenesis. It is demonstrated that cytokine levels changein combination with increment of epileptic activity in rats whichare genetic models of absence epilepsy (WAG/Rij). In our study,the required experimental setup is prepared to investigate poly-morphism of IL-1�-511C > T(rs16944), IL-6-572C > G(rs1800795),IL-10-592A > C(rs1800872), TNF�-308G > A(rs1800629) associatedwith epilepsy in human on absence epileptic WAG/Rij rats (n = 20)and non-epileptic Wistar rats (n = 20). Thus specified humansingle nucleotide polymorphisms (SNPs) sequence regions weredetermined in NCBI SNP database. Same sequence regions weredetermined by Ensemble database and sequences aligned at NCBIblast. Rat primers designed for four SNP sequence regions. SNPregions were amplified by PCR and analysed with Single StrandConformation Polymorphism (SSCP). According to our preliminaryresults we can conclude that human three SNPs which were associ-ated with epilepsy, are not responsible for development of epilepsyin rats. SSCP analysis for TNF� different band patterns is shown butnot every individual.

http://dx.doi.org/10.1016/j.jbiotec.2014.07.303

Integrated high throughput analysis identifiesAP-1 as a crucial factor of p53-mediated cellfate decision

Hai Li 1,∗, Yu Zhang 2, Galina Selivanova 1

1 Department of Microbiology and Tumor Biology,Karolinska Institutet, Stockholm, Sweden2 College of Life Science, Northeast AgriculturalUniversity, Harbin, PR China

E-mail address: hailichina@gmail.com (H. Li).

Reactivation of p53 is a promising therapeutic strategy againsthuman cancers. Many efforts have been made to develop p53-reactivating drugs. However, the biological outcomes of p53restoration are various, raging from cell cycle arrest to apopto-

sis, and hard to predict. To succeed in p53-based cancer therapies,deeper understanding of the mechanism of p53-mediated cellfate decision is needed. Therefore, we performed bioinformat-ics analysis of gene expression profiles (microarray) combinedwith genome-wide analysis of p53 binding sites (ChIP-seq) usingMCF-7 breast carcinoma cell line treated with two p53 activators,RITA (inducing p53-dependent apoptosis) and Nutlin (inducingp53-dependent cell cycle arrest), as a model. Comparison of thetranscription profiles revealed a group of genes differently regu-lated by RITA and Nutlin. Further analysis of gene expressions incombination with distribution of p53 binding sites across genomeshowed 21% of these genes were p53 direct targets. Protein–proteininteraction analysis of these p53 targets suggested AP-1 as a crucialfactor in p53-mediated cell fate decision, which was confirmed byfunctional studies in cells expressing shRNAs for Jun or Fos. Fur-ther characterization of the signaling pathways co-regulated byp53 and AP-1 may contribute to the discovery of novel therapeuticapproaches.

http://dx.doi.org/10.1016/j.jbiotec.2014.07.304

Development of electrospun adhesive layercontaining ciprofloxacin/coconut oil for use asantibacterial wound dressing

Sonthaya Chaiarwut 1,∗, Pitt Supaphol 1, PrasitPavasant 2

1 The Petroleum and Petrochemical College,Chulalongkorn University, Bangkok, Thailand2 Department of Dentistry, ChulalongkornUniversity, Bangkok, Thailand

E-mail address: tonh2ook@gmail.com (S. Chaiarwut).

Poly(vinyl acetate), a non-toxic adhesive, was used to prepareelectrospun fibrous membranes by blending an antibiotic andwound healing supporter to cover commercial polyurethane (PU)substrate. PU film has many advantages such as excellent air per-meation and good mechanical properties. This study focused onthe synergistic interaction between coconut oil and Ciprofloxacin(CPF), which can be toxic to the growth of cells. The surface mor-phology, drug release, and mechanical properties of tissue adhesivewere characterized by scanning electron microscopy (SEM), UV–visspectroscopy and the T-Peel test (ASTM D6252). The antibacterialactivity was tested against pathogenic bacteria, Escherichia coli andStaphylococcus aureus which are the representatives of gram(−)and gram(+) bacteria, respectively. The inhibition zones of E. coliand S. aureus appeared when CPF and coconut oil was loaded at1.25 mg/ml and 0.5 ml, respectively. For the cytotoxicity of cell via-bility, the mats were evaluated with mouse fibroblast (L929) andhuman fibroblast cells by culturing on the wound dressing surfaceto determine indirect cytotoxicity using MTT essay and then thetesting of mats showed good results for cell viability of both L929and human fibroblast cells.

http://dx.doi.org/10.1016/j.jbiotec.2014.07.305