- Email: [email protected]
A novel approach of interaction between bilirubin and human serum albumin in the presence of Aspirine
S294
Abstracts
reduce death and maintaining extensive levels on cell viability under serum free media in cell culture are important subjects in production of recombinant proteins. Insulin-Like Growth Factor-I (IGF-I) has mitogenic and antiapoptotic activity protecting cultures from diverse death inducing stimuli. Methods: We studied the effect of different concentrations of insulin- like growth factor I (IGF-I) on anchorage dependent CHO–K1 cell line for 24, 48 h. Cells were cultivated in DMEM supplemented with 10% fetal bovine serum (FBS) and 1% penicillin and streptomycin antibiotic. Serum was removed and 10–50 ng/mL IGF-I was added. Cells were stained with trypan blue and counted in hemocytometer. Cell viability was determined by MTT assay. Apoptosis was assessed by caspase 3 detection kit and cells morphologic were examined with inverted microscope. Results: In growing cells at the presence of IGF-I cell number above control levels in different times increased. Proliferation and viability increased in dose-dependent manner as IGF-I was increased. Conclusion: These investigations demonstrated that IGF-I can act as survival factor, maintaining viable cell density for an extended period of time and antiapoptotic factor, suppressing apoptosis in serum-free cultures of CHO cells. Keywords: Insulin– like growth factor-I, Apoptosis, Chinese Hamster Ovary cells, Cell culture doi:10.1016/j.clinbiochem.2011.08.738
Poster – [A-10-788-1] Identifying likely causal connections between gene expression levels using a Mendelian randomization approach Hanieh Yaghootkara, Dena Hernandezb, Michael Nallsb, Andrew Wooda, Raphael Gibbsb, Lorna Harriesc, Sean Chongb, Matthew Mooreb, Jack Guralnikd, Stefaniai Bandinelle, Anna Murraya, Luigi Ferruccia, Andrew Singletonc, David Melzera, Timothy Fraylinga a University of Exeter, Exeter EX1 2LU, UK b Building 35, Room 1A1014, 35 Lincoln Drive, Bethesda, MD 20892, USA c University of Exeter, Barrack Road, Exeter, EX2 5DW, UK d Baltimore, MD, USA e Florence, Italy E-mail addresses: [email protected] (H. Yaghootkar), [email protected] (D. Hernandez), [email protected] (M. Nalls), [email protected] (A. Wood), [email protected] (R. Gibbs), [email protected] (L. Harries), [email protected] (S. Chong), [email protected] (M. Moore), [email protected] (J. Guralnik), [email protected] (S. Bandinell), [email protected] (A. Murray), [email protected] (L. Ferrucci), [email protected] (A. Singleton), [email protected] (D. Melzer), [email protected] (T. Frayling) Introduction: Gene expression studies identify networks of correlated genes that are biologically difficult to interpret. The direction of which gene controls which, or whether a third gene, or environmental factors, or any combination of these possibilities, are influencing the correlation is unknown. In the present study, we aimed to use genetic variants (cis eQTLs) that are significantly associated with expression levels of their neighbor genes to find the direction, if causal, of genes association by following the principle of Mendelian randomization. Methods: We used one of the largest eQTL datasets described to date—1437 eQTLs identified in expression data from fresh lympho-
cytes from >600 individuals in the InCHIANTI study. Instrumental variable analysis, a statistical method to assess causality, was run on 3487 pairs of genes with correlation ≥ 0.5. Results: We found evidence for causal connections between 455 gene pairs. By looking at the causal regulators, we identified 87 “master” genes controlling at least two other genes; 106 “slave” genes controlled by more than one other genes, and 26 pairs of “gladiator” genes with evidence of reciprocal control. Four regulator genes were linked to four already known human diseases/traits associated loci, including type 2 diabetes, prostate cancer, height and lipid metabolites. Conclusion: Combining eQTLs data with principle of Mendelian randomization is starting to allow us to assess causality in gene regulation networks, and potentially understand the mechanism of diseases. Keywords: Gene expression, Genetic variants, eQTL doi:10.1016/j.clinbiochem.2011.08.739
Poster – [A-10-793-1] A novel approach of interaction between bilirubin and human serum albumin in the presence of Aspirine Akram Hossenzadeha, Jamshid Chamania, Mohsen Gharanfolib a Department of Biology, Faculty of Sciences, Islamic Azad University, Mashhad Branch, Mashhad, Iran b Department of Development Biology, Culture and Science University, Tehran, Iran E-mail addresses: [email protected] (A. Hossenzadeh), [email protected] (J. Chamani), [email protected] (M. Gharanfoli) Introduction: Bilirubin (BR), the yellow neurotoxic pigment of jaundice and the end product of heme metabolism in mammals, is lipophilic tetrapyrrole dicarboxylic acid. Human serum albumin (HSA) is a principal extra cellular protein. Its primary pharmacokinetics function is participating in absorption, distribution, metabolism and excretion of drug. Acetylsalicylic acid (ASA) has been shown to reduce the risk for colorectal cancer by as much as approximately 40%, a property that is shared by other nonsteroidal anti-inflammatory drugs. Materials and methods: BR, ASA and HSA were purchased from Sigma. Fluorescence measurements were carried out on a F-2500. Molecular modeling was done by autodock software. Result and discussion: Investigating the interaction of drugs to HSA can elucidate the properties of drug–protein complex. Synchronous fluorescence spectroscopy from the interaction of HSA with BR and in the presence of Aspirin showed that ligand affect the conformation of HSA. Interaction of BR and ASA with HSA changed the binding affinity of second drug that can use the location in the binding site on protein. On the other hand we found that the binding affinity of ASA bound to HAS was stronger in the presence of BR, and considered the same result for addition of ASA to HAS-BR complex. Keywords: Bilirubin, Aspirin, Human serum albumin, Molecular modeling doi:10.1016/j.clinbiochem.2011.08.740
Poster – [A-10-793-3] Applying the three-dimensional fluorescence spectroscopy for studying the interaction between bilirubin and human serum albumin as binary system Akram Hosenzadeha, Mohsen Gharanfolib, Jamshid Chamania, Masomeh Vahidzadeha
Abstracts
reduce death and maintaining extensive levels on cell viability under serum free media in cell culture are important subjects in production of recombinant proteins. Insulin-Like Growth Factor-I (IGF-I) has mitogenic and antiapoptotic activity protecting cultures from diverse death inducing stimuli. Methods: We studied the effect of different concentrations of insulin- like growth factor I (IGF-I) on anchorage dependent CHO–K1 cell line for 24, 48 h. Cells were cultivated in DMEM supplemented with 10% fetal bovine serum (FBS) and 1% penicillin and streptomycin antibiotic. Serum was removed and 10–50 ng/mL IGF-I was added. Cells were stained with trypan blue and counted in hemocytometer. Cell viability was determined by MTT assay. Apoptosis was assessed by caspase 3 detection kit and cells morphologic were examined with inverted microscope. Results: In growing cells at the presence of IGF-I cell number above control levels in different times increased. Proliferation and viability increased in dose-dependent manner as IGF-I was increased. Conclusion: These investigations demonstrated that IGF-I can act as survival factor, maintaining viable cell density for an extended period of time and antiapoptotic factor, suppressing apoptosis in serum-free cultures of CHO cells. Keywords: Insulin– like growth factor-I, Apoptosis, Chinese Hamster Ovary cells, Cell culture doi:10.1016/j.clinbiochem.2011.08.738
Poster – [A-10-788-1] Identifying likely causal connections between gene expression levels using a Mendelian randomization approach Hanieh Yaghootkara, Dena Hernandezb, Michael Nallsb, Andrew Wooda, Raphael Gibbsb, Lorna Harriesc, Sean Chongb, Matthew Mooreb, Jack Guralnikd, Stefaniai Bandinelle, Anna Murraya, Luigi Ferruccia, Andrew Singletonc, David Melzera, Timothy Fraylinga a University of Exeter, Exeter EX1 2LU, UK b Building 35, Room 1A1014, 35 Lincoln Drive, Bethesda, MD 20892, USA c University of Exeter, Barrack Road, Exeter, EX2 5DW, UK d Baltimore, MD, USA e Florence, Italy E-mail addresses: [email protected] (H. Yaghootkar), [email protected] (D. Hernandez), [email protected] (M. Nalls), [email protected] (A. Wood), [email protected] (R. Gibbs), [email protected] (L. Harries), [email protected] (S. Chong), [email protected] (M. Moore), [email protected] (J. Guralnik), [email protected] (S. Bandinell), [email protected] (A. Murray), [email protected] (L. Ferrucci), [email protected] (A. Singleton), [email protected] (D. Melzer), [email protected] (T. Frayling) Introduction: Gene expression studies identify networks of correlated genes that are biologically difficult to interpret. The direction of which gene controls which, or whether a third gene, or environmental factors, or any combination of these possibilities, are influencing the correlation is unknown. In the present study, we aimed to use genetic variants (cis eQTLs) that are significantly associated with expression levels of their neighbor genes to find the direction, if causal, of genes association by following the principle of Mendelian randomization. Methods: We used one of the largest eQTL datasets described to date—1437 eQTLs identified in expression data from fresh lympho-
cytes from >600 individuals in the InCHIANTI study. Instrumental variable analysis, a statistical method to assess causality, was run on 3487 pairs of genes with correlation ≥ 0.5. Results: We found evidence for causal connections between 455 gene pairs. By looking at the causal regulators, we identified 87 “master” genes controlling at least two other genes; 106 “slave” genes controlled by more than one other genes, and 26 pairs of “gladiator” genes with evidence of reciprocal control. Four regulator genes were linked to four already known human diseases/traits associated loci, including type 2 diabetes, prostate cancer, height and lipid metabolites. Conclusion: Combining eQTLs data with principle of Mendelian randomization is starting to allow us to assess causality in gene regulation networks, and potentially understand the mechanism of diseases. Keywords: Gene expression, Genetic variants, eQTL doi:10.1016/j.clinbiochem.2011.08.739
Poster – [A-10-793-1] A novel approach of interaction between bilirubin and human serum albumin in the presence of Aspirine Akram Hossenzadeha, Jamshid Chamania, Mohsen Gharanfolib a Department of Biology, Faculty of Sciences, Islamic Azad University, Mashhad Branch, Mashhad, Iran b Department of Development Biology, Culture and Science University, Tehran, Iran E-mail addresses: [email protected] (A. Hossenzadeh), [email protected] (J. Chamani), [email protected] (M. Gharanfoli) Introduction: Bilirubin (BR), the yellow neurotoxic pigment of jaundice and the end product of heme metabolism in mammals, is lipophilic tetrapyrrole dicarboxylic acid. Human serum albumin (HSA) is a principal extra cellular protein. Its primary pharmacokinetics function is participating in absorption, distribution, metabolism and excretion of drug. Acetylsalicylic acid (ASA) has been shown to reduce the risk for colorectal cancer by as much as approximately 40%, a property that is shared by other nonsteroidal anti-inflammatory drugs. Materials and methods: BR, ASA and HSA were purchased from Sigma. Fluorescence measurements were carried out on a F-2500. Molecular modeling was done by autodock software. Result and discussion: Investigating the interaction of drugs to HSA can elucidate the properties of drug–protein complex. Synchronous fluorescence spectroscopy from the interaction of HSA with BR and in the presence of Aspirin showed that ligand affect the conformation of HSA. Interaction of BR and ASA with HSA changed the binding affinity of second drug that can use the location in the binding site on protein. On the other hand we found that the binding affinity of ASA bound to HAS was stronger in the presence of BR, and considered the same result for addition of ASA to HAS-BR complex. Keywords: Bilirubin, Aspirin, Human serum albumin, Molecular modeling doi:10.1016/j.clinbiochem.2011.08.740
Poster – [A-10-793-3] Applying the three-dimensional fluorescence spectroscopy for studying the interaction between bilirubin and human serum albumin as binary system Akram Hosenzadeha, Mohsen Gharanfolib, Jamshid Chamania, Masomeh Vahidzadeha