- Email: [email protected]
Pro-oxidant–antioxidant balance in obese patients undergoing acupuncture
Abstracts
structures. In other words G-quadruplex structure is stable in the presence of hemin. Keywords: DNAzyme, Hemin, Peroxidase, Circular dichroism doi:10.1016/j.clinbiochem.2011.08.215
Molecular Biology and Genetic Disorders E Poster – [A-10-841-1] Association of promoter variation in HMOX1 and UGT1A1 genes with Fabry disease Alena Jiraskova, Jaglika Jovanovska, Libor Vitek Charles Univ., Prague, Czech Republic E-mail address: [email protected] (L. Vitek) Introduction: Heme oxygenase-1 (encoded by HMOX1) and bilirubin UDP-glucuronosyltransferase (UGT1A1) play an important role in the oxidative stress defence. Microsatellite variations in the promoter regions modulate HMOX1 and UGT1A1 gene expression and are associated with oxidative stress-mediated diseases, such as atherosclerosis or certain cancers. The objective of this study was to determine whether these variations affect development of Fabry disease (FD), a metabolic disorder of glycosphingolipid metabolism caused by deficiency of the lysosomal hydrolase α-galactosidase A resulting in accumulation of glycosphingolipids in various tissues. Materials and methods: The study was performed on 51 FD patients (M:F ratio = 0.54) and 89 healthy age- and sex-matched controls. The (GT)n and (TA)n dinucleotide variations were determined by fragment analysis using an automated capillary DNA sequencer. The length variations of (GT)n repeats were classified into short S (n < 27), medium M (n = 27–32) and long L (n ≥ 33) subgroups. Logistic regression analysis was used to compare the frequency of the most important alleles in FD and control groups. Results: No significant differences in frequencies of L-allele in HMOX1 (associated with lower enzyme activity) between FD and control groups were found (OR = 1.69; 95%CI = 0.54–5.24, p = 0.46). In contrast, frequency of TA7 allele responsible for higher serum bilirubin levels was associated with decreased odds for FD manifestation (OR = 0.43, 95%CI = 0.20–0.92, p = 0.018), which was more pronounced in females (OR = 0.32, 95%CI = 0.12–0.86, p = 0.01). Conclusions: Results of our study suggest that (TA)n repeat variations in UGT1A1 provide protection against FD, whereas (GT)n variations in HMOX1 are likely not to contribute to the risk of FD. Keywords: Heme oxygenase, UGT1A1, Bilirubin, Polymorphisms, Microsatellite variations, Fabry disease doi:10.1016/j.clinbiochem.2011.08.216
E Poster – [A-10-1094-1] Association Between Neuropeptide Y LEU7PRO Polymorphism and Metabolic Syndrome in Patients with Coronary Artery Disease Masoudi Kazemabad Alia, Ghayour Mobarhan Majidb, Jamialahmadi Khadijehc, moohebati Mohsend, Mojarrad Majide, Eidi Akrama, Khojasteh Roshanakb, Mousavib, Dehghan Rahelehc, Paydar Roghaieb a Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran b Biochemical and Nutritional Research Centre, Mashhad University of Medical Sciences, Mashhad, Iran c Department of Modern Sciences and Technologies, Faculty of Medicine, Azadi Square, Mashhad, Iran
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d Department of Cardiology and Cardiovascular Research Centre, Ghaem Hospital, Ahmadabad Street, Mashhad, Iran e Department of Genetic, Faculty of Medicine, Azadi Square, Mashhad, Iran E-mail addresses: [email protected] (M. K. Ali), [email protected] (J. Khadijeh), [email protected] (M. Mohsen), [email protected] (M. Majid), [email protected] (K. Roshanak), [email protected] (D. Raheleh), [email protected] (P. Roghaie)
Introduction: Metabolic syndrome (MS) is a combination of medical disorders that increase the risk of developing cardiovascular disease. Neuropeptide Y acts directly in the periphery on fat tissue and mediates metabolic syndrome. However, there is a little data about NPY gene polymorphism and MS in Iran. Therefore, we wish to investigate the relationship between NPY Leu7Pro polymorphism and MS in Iran. Materials & Methods: A total of 550 patients with coronary artery disease were recruited; 184 patients with metabolic syndrome and 366 had not metabolic syndrome from department of Cardiology, Ghaem hospital, Mashhad, Iran. Samples DNA was extracted in salting out method and the Leu7Pro polymorphism analysis was performed with PCR-RFLP technique for all subjects. Results: A significantly higher frequency in Leu7Pro was observed in metabolic syndrome patients compared with the non-metabolic syndrome patients (p< 0.01). Conclusion: The Leu7Pro polymorphism was associated with metabolic syndrome in Iranian northeastern population. Keywords: Metabolic syndrome, Neuropeptide Y, Leu7Pro polymorphism, PCR-RFLP
doi:10.1016/j.clinbiochem.2011.08.217
Modern Techniques and Biomedical Diagnosis E Poster – [A-10-4-1] Pro-oxidant–antioxidant balance in obese patients undergoing acupuncture Abbasi Parizad Parisa, Majid Ghayour-Mobarhan, Parizadeh Mohammad Reza, Hamid Abdi Biochemical and Nutritional Research Centre, Mashhad University of Medical Sciences, Mashhad, Iran E-mail addresses: [email protected] (A.P. Parisa), [email protected] (M. Ghayour-Mobarhan), [email protected] (P.M. Reza), [email protected] (H. Abdi) Introduction: There are limited data that reported on the effect of acupuncture on the pro-oxidant–antioxidant status in obese patients. In this study we evaluated the effects of body acupuncture, as a kind of complementary treatment, with low calorie diet on losing weight and measuring the serum levels of PAB. Methods: 94 healthy obese volunteers with Body Mass Indexes (BMI) above (than) 25 kg/m2 were divided case per case into two groups as Real(1) and Sham(2); 1) Real acupuncture group (n = 52) treating with real electroacupuncture and diet control and 2) the unreal acupuncture group (n = 42) treating with sham electroacupuncture and diet control. All subjects in both groups received lowcalorie diet for six weeks and two sessions of body acupuncture per week. Body weight (BW), BMI, waist circumference (WC) and serum levels of PAB were measured in the first day of the trial, at 45 days and after 90 days after the treatment.
S96
Abstracts
Results: A significant reduction was observed in BW and BMI in case groups(P≤0.05), and in WC(P≤0.01). However, differences between the percentage of reductions in neither BW nor BMI and WC of subjects in the case group and those in the control group were not statistically significant. No significant changes were observed in PAB value in both groups. Conclusion: Electrobody acupuncture could be effective on reducing weight, although, it doesn't decrease the balance of antioxidant–oxidants in the obese patients, but have shown at least no side effect in diminishing weight. Keywords: Obesity, Acupuncture, Pro-oxidant–antioxidant balance (PAB) doi:10.1016/j.clinbiochem.2011.08.218
E Poster – [A-10-40-1] Mitogen activated protein kinases in human peripheral nerve protein phosphorylation: Monoclonal antibody mediated inhibition assay Mona Latifi, Hossein Mehraban Islamic Azad University, Jahrom Branch, Jahrom, Iran E-mail addresses: [email protected] (M. Latifi), [email protected] (H. Mehraban) Introduction: The amount of biochemicals in the brain decreases with age. Neurodegeneration disorders are the result of massively accelerated rates of synaptic degeneration and neural cell loss. Some of these disorders are common and are thought to be caused by environmental factors and genetic susceptibility along with a small percentage due to inherited forms of the disease (e.g. Alzheimer disease). The MAP kinases and phosphatidylinositol-3 kinase (PI3K) are serine/ threonine protein kinases that play critical role in neuronal growth, differentiation and survival. In general, activation of the extra cellular signal-regulated kinase members of MAP kinase family (ERK or P42/P44 MAP kinases) and PI3K signaling pathway promote cell survival, while members of MAP kinase family known as the stress activated protein kinases (JNK) and the p38 MAP kinase, promote cell death. The present study evaluated the effect of blocking specific proteins/kinases of this pathway observing the phosphorylation of substrate proteins. Material and methods: We studied this effect on a well characterized system, the peripheral nervous system. Protein phosphorylation was carried out with or without the presence of specific monoclonal antibodies. The reaction mixtures were subjected to SDS PAGE and autoradiography. Results and conclusion: The result showed that blocking of JNK and p38 MAP kinases did not significantly affect the overall phosphorylation but, ERK MAP kinase blocking resulted in significant decrease in the phosphorylation of major peripheral nerve protein p0. Diseases caused by MAPK protein alteration are Huntington disease, Parkinson disease, cancer, etc. Our observation bears significance in designing molecules to regulate, substitute or bypass these defects.
In this report we used gold magnetic nanoparticles (GMN) not only to covalently attach an antibody using a spacer arm, but also to chemically adsorb an antibody without any spacer arm. In chemical adsorption, Anti IgG_Ab was directly attached to surface of GMN by passive adsorption. Whereas in covalent binding Anti IgG_Ab, through a self assembled monolayer (SAM) of 11-mercaptoundecanoic acid (MUA) via carbodiimide coupling reaction using N-(3-dimethylaminopropyl)-N′-ethyl carbodiimide hydrochloride (EDC) and N-Hydroxy Succinimide (NHS) was immobilized on gold magnetic nanoparticle. The GMN/Ab1 modified gold electrode was used for a sandwich electrochemical immunoassay. The detection was done using cyclic voltammetric measurements using horse radish peroxidase (HRP) as enzyme label on secondary antibody (Ab2). The chemical adsorption showed higher formal potential and peak to peak separation compared with covalent binding. Value of formal potential (E0, the average of the cathodic and anodic peak potential) were 738.4 mV and 698 mV, respectively, and peak to peak separation (ΔEp=Epa–Epc) were 653.2 mV and 617.2 mV, respectively. Therefore, we anticipate that the immobilization based on chemical adsorption of anti-IgG would be the most suitable for human IgG detection. Keywords: Antibody, Cyclic voltammetry, Covalent binding, Chemical adsorption, Gold magnetic nanoparticle doi:10.1016/j.clinbiochem.2011.08.220
E Poster – [A-10-254-2] Immobilized horseradish peroxidase on Nafion-cysteine modified gold electrode as a biosensor for hydrogen peroxide Hedayatollah Ghourchian, Jun Hong, Faezeh Hamidi Institute if Biochemistry and Biophysics, Tehran University, Tehran, Iran E-mail address: [email protected] (H. Ghourchian) Introduction: Horseradish peroxidase was immobilized on a newly designed functional membrane composed of nanoparticles of methylene blue and Nafion perfluorosulfonated ion-exchange resin (Nafionw) to develop a biosensor for determination of hydrogen peroxide. Method: At first the membrane was constructed and characterized by the methods of scanning electron microscope, transmission electron microscope, UV–Vis, and Fourier transform infrared. The membrane at glassy carbon electrode was directly applicable for determining H2O2 concentration. In the next step, horseradish peroxidase was immobilized on the MB-Nafion functional membrane, which showed a quasireversible electrochemical behavior with a formal potential of 2296 mV. Results: The resulting biosensor exhibited a good stability and higher sensitivity to H2O2 relative to the membrane without horseradish perioxidase. Conclusion: The linear range of this biosensor for H2O2 determination was from 2 ×10− 5 to 7× 10− 3 M, with a detection limit of 7× 10− 6 M. Keywords: Methylene blue, Nafion, Functional membrane, Biosensor, HRP, Hydrogen peroxide
Keywords: MAP kinase, Monoclonal antibody, Protein p0 doi:10.1016/j.clinbiochem.2011.08.221 doi:10.1016/j.clinbiochem.2011.08.219
E Poster – [A-10-198-1] Electrochemical study of the function of antibodies immobilized through chemical absorption and covalent binding Sara Nourani, Khadijeh Eskandari, Hedayatolah Ghourchian Tehran University, Tehran, Iran E-mail addresses: [email protected] (S. Nourani), [email protected] (K. Eskandari), [email protected] (H. Ghourchian)
E Poster – [A-10-303-1] Nano-light technology-based probe, a tool for detection of apoptotic cells Mahboobeh Nazaria, Rahman Emamzadehb, Saman Hossinkhania a Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran b Department of Biology, Faculty of Science, University of Isfahan, Tehran, Iran
structures. In other words G-quadruplex structure is stable in the presence of hemin. Keywords: DNAzyme, Hemin, Peroxidase, Circular dichroism doi:10.1016/j.clinbiochem.2011.08.215
Molecular Biology and Genetic Disorders E Poster – [A-10-841-1] Association of promoter variation in HMOX1 and UGT1A1 genes with Fabry disease Alena Jiraskova, Jaglika Jovanovska, Libor Vitek Charles Univ., Prague, Czech Republic E-mail address: [email protected] (L. Vitek) Introduction: Heme oxygenase-1 (encoded by HMOX1) and bilirubin UDP-glucuronosyltransferase (UGT1A1) play an important role in the oxidative stress defence. Microsatellite variations in the promoter regions modulate HMOX1 and UGT1A1 gene expression and are associated with oxidative stress-mediated diseases, such as atherosclerosis or certain cancers. The objective of this study was to determine whether these variations affect development of Fabry disease (FD), a metabolic disorder of glycosphingolipid metabolism caused by deficiency of the lysosomal hydrolase α-galactosidase A resulting in accumulation of glycosphingolipids in various tissues. Materials and methods: The study was performed on 51 FD patients (M:F ratio = 0.54) and 89 healthy age- and sex-matched controls. The (GT)n and (TA)n dinucleotide variations were determined by fragment analysis using an automated capillary DNA sequencer. The length variations of (GT)n repeats were classified into short S (n < 27), medium M (n = 27–32) and long L (n ≥ 33) subgroups. Logistic regression analysis was used to compare the frequency of the most important alleles in FD and control groups. Results: No significant differences in frequencies of L-allele in HMOX1 (associated with lower enzyme activity) between FD and control groups were found (OR = 1.69; 95%CI = 0.54–5.24, p = 0.46). In contrast, frequency of TA7 allele responsible for higher serum bilirubin levels was associated with decreased odds for FD manifestation (OR = 0.43, 95%CI = 0.20–0.92, p = 0.018), which was more pronounced in females (OR = 0.32, 95%CI = 0.12–0.86, p = 0.01). Conclusions: Results of our study suggest that (TA)n repeat variations in UGT1A1 provide protection against FD, whereas (GT)n variations in HMOX1 are likely not to contribute to the risk of FD. Keywords: Heme oxygenase, UGT1A1, Bilirubin, Polymorphisms, Microsatellite variations, Fabry disease doi:10.1016/j.clinbiochem.2011.08.216
E Poster – [A-10-1094-1] Association Between Neuropeptide Y LEU7PRO Polymorphism and Metabolic Syndrome in Patients with Coronary Artery Disease Masoudi Kazemabad Alia, Ghayour Mobarhan Majidb, Jamialahmadi Khadijehc, moohebati Mohsend, Mojarrad Majide, Eidi Akrama, Khojasteh Roshanakb, Mousavib, Dehghan Rahelehc, Paydar Roghaieb a Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran b Biochemical and Nutritional Research Centre, Mashhad University of Medical Sciences, Mashhad, Iran c Department of Modern Sciences and Technologies, Faculty of Medicine, Azadi Square, Mashhad, Iran
S95
d Department of Cardiology and Cardiovascular Research Centre, Ghaem Hospital, Ahmadabad Street, Mashhad, Iran e Department of Genetic, Faculty of Medicine, Azadi Square, Mashhad, Iran E-mail addresses: [email protected] (M. K. Ali), [email protected] (J. Khadijeh), [email protected] (M. Mohsen), [email protected] (M. Majid), [email protected] (K. Roshanak), [email protected] (D. Raheleh), [email protected] (P. Roghaie)
Introduction: Metabolic syndrome (MS) is a combination of medical disorders that increase the risk of developing cardiovascular disease. Neuropeptide Y acts directly in the periphery on fat tissue and mediates metabolic syndrome. However, there is a little data about NPY gene polymorphism and MS in Iran. Therefore, we wish to investigate the relationship between NPY Leu7Pro polymorphism and MS in Iran. Materials & Methods: A total of 550 patients with coronary artery disease were recruited; 184 patients with metabolic syndrome and 366 had not metabolic syndrome from department of Cardiology, Ghaem hospital, Mashhad, Iran. Samples DNA was extracted in salting out method and the Leu7Pro polymorphism analysis was performed with PCR-RFLP technique for all subjects. Results: A significantly higher frequency in Leu7Pro was observed in metabolic syndrome patients compared with the non-metabolic syndrome patients (p< 0.01). Conclusion: The Leu7Pro polymorphism was associated with metabolic syndrome in Iranian northeastern population. Keywords: Metabolic syndrome, Neuropeptide Y, Leu7Pro polymorphism, PCR-RFLP
doi:10.1016/j.clinbiochem.2011.08.217
Modern Techniques and Biomedical Diagnosis E Poster – [A-10-4-1] Pro-oxidant–antioxidant balance in obese patients undergoing acupuncture Abbasi Parizad Parisa, Majid Ghayour-Mobarhan, Parizadeh Mohammad Reza, Hamid Abdi Biochemical and Nutritional Research Centre, Mashhad University of Medical Sciences, Mashhad, Iran E-mail addresses: [email protected] (A.P. Parisa), [email protected] (M. Ghayour-Mobarhan), [email protected] (P.M. Reza), [email protected] (H. Abdi) Introduction: There are limited data that reported on the effect of acupuncture on the pro-oxidant–antioxidant status in obese patients. In this study we evaluated the effects of body acupuncture, as a kind of complementary treatment, with low calorie diet on losing weight and measuring the serum levels of PAB. Methods: 94 healthy obese volunteers with Body Mass Indexes (BMI) above (than) 25 kg/m2 were divided case per case into two groups as Real(1) and Sham(2); 1) Real acupuncture group (n = 52) treating with real electroacupuncture and diet control and 2) the unreal acupuncture group (n = 42) treating with sham electroacupuncture and diet control. All subjects in both groups received lowcalorie diet for six weeks and two sessions of body acupuncture per week. Body weight (BW), BMI, waist circumference (WC) and serum levels of PAB were measured in the first day of the trial, at 45 days and after 90 days after the treatment.
S96
Abstracts
Results: A significant reduction was observed in BW and BMI in case groups(P≤0.05), and in WC(P≤0.01). However, differences between the percentage of reductions in neither BW nor BMI and WC of subjects in the case group and those in the control group were not statistically significant. No significant changes were observed in PAB value in both groups. Conclusion: Electrobody acupuncture could be effective on reducing weight, although, it doesn't decrease the balance of antioxidant–oxidants in the obese patients, but have shown at least no side effect in diminishing weight. Keywords: Obesity, Acupuncture, Pro-oxidant–antioxidant balance (PAB) doi:10.1016/j.clinbiochem.2011.08.218
E Poster – [A-10-40-1] Mitogen activated protein kinases in human peripheral nerve protein phosphorylation: Monoclonal antibody mediated inhibition assay Mona Latifi, Hossein Mehraban Islamic Azad University, Jahrom Branch, Jahrom, Iran E-mail addresses: [email protected] (M. Latifi), [email protected] (H. Mehraban) Introduction: The amount of biochemicals in the brain decreases with age. Neurodegeneration disorders are the result of massively accelerated rates of synaptic degeneration and neural cell loss. Some of these disorders are common and are thought to be caused by environmental factors and genetic susceptibility along with a small percentage due to inherited forms of the disease (e.g. Alzheimer disease). The MAP kinases and phosphatidylinositol-3 kinase (PI3K) are serine/ threonine protein kinases that play critical role in neuronal growth, differentiation and survival. In general, activation of the extra cellular signal-regulated kinase members of MAP kinase family (ERK or P42/P44 MAP kinases) and PI3K signaling pathway promote cell survival, while members of MAP kinase family known as the stress activated protein kinases (JNK) and the p38 MAP kinase, promote cell death. The present study evaluated the effect of blocking specific proteins/kinases of this pathway observing the phosphorylation of substrate proteins. Material and methods: We studied this effect on a well characterized system, the peripheral nervous system. Protein phosphorylation was carried out with or without the presence of specific monoclonal antibodies. The reaction mixtures were subjected to SDS PAGE and autoradiography. Results and conclusion: The result showed that blocking of JNK and p38 MAP kinases did not significantly affect the overall phosphorylation but, ERK MAP kinase blocking resulted in significant decrease in the phosphorylation of major peripheral nerve protein p0. Diseases caused by MAPK protein alteration are Huntington disease, Parkinson disease, cancer, etc. Our observation bears significance in designing molecules to regulate, substitute or bypass these defects.
In this report we used gold magnetic nanoparticles (GMN) not only to covalently attach an antibody using a spacer arm, but also to chemically adsorb an antibody without any spacer arm. In chemical adsorption, Anti IgG_Ab was directly attached to surface of GMN by passive adsorption. Whereas in covalent binding Anti IgG_Ab, through a self assembled monolayer (SAM) of 11-mercaptoundecanoic acid (MUA) via carbodiimide coupling reaction using N-(3-dimethylaminopropyl)-N′-ethyl carbodiimide hydrochloride (EDC) and N-Hydroxy Succinimide (NHS) was immobilized on gold magnetic nanoparticle. The GMN/Ab1 modified gold electrode was used for a sandwich electrochemical immunoassay. The detection was done using cyclic voltammetric measurements using horse radish peroxidase (HRP) as enzyme label on secondary antibody (Ab2). The chemical adsorption showed higher formal potential and peak to peak separation compared with covalent binding. Value of formal potential (E0, the average of the cathodic and anodic peak potential) were 738.4 mV and 698 mV, respectively, and peak to peak separation (ΔEp=Epa–Epc) were 653.2 mV and 617.2 mV, respectively. Therefore, we anticipate that the immobilization based on chemical adsorption of anti-IgG would be the most suitable for human IgG detection. Keywords: Antibody, Cyclic voltammetry, Covalent binding, Chemical adsorption, Gold magnetic nanoparticle doi:10.1016/j.clinbiochem.2011.08.220
E Poster – [A-10-254-2] Immobilized horseradish peroxidase on Nafion-cysteine modified gold electrode as a biosensor for hydrogen peroxide Hedayatollah Ghourchian, Jun Hong, Faezeh Hamidi Institute if Biochemistry and Biophysics, Tehran University, Tehran, Iran E-mail address: [email protected] (H. Ghourchian) Introduction: Horseradish peroxidase was immobilized on a newly designed functional membrane composed of nanoparticles of methylene blue and Nafion perfluorosulfonated ion-exchange resin (Nafionw) to develop a biosensor for determination of hydrogen peroxide. Method: At first the membrane was constructed and characterized by the methods of scanning electron microscope, transmission electron microscope, UV–Vis, and Fourier transform infrared. The membrane at glassy carbon electrode was directly applicable for determining H2O2 concentration. In the next step, horseradish peroxidase was immobilized on the MB-Nafion functional membrane, which showed a quasireversible electrochemical behavior with a formal potential of 2296 mV. Results: The resulting biosensor exhibited a good stability and higher sensitivity to H2O2 relative to the membrane without horseradish perioxidase. Conclusion: The linear range of this biosensor for H2O2 determination was from 2 ×10− 5 to 7× 10− 3 M, with a detection limit of 7× 10− 6 M. Keywords: Methylene blue, Nafion, Functional membrane, Biosensor, HRP, Hydrogen peroxide
Keywords: MAP kinase, Monoclonal antibody, Protein p0 doi:10.1016/j.clinbiochem.2011.08.221 doi:10.1016/j.clinbiochem.2011.08.219
E Poster – [A-10-198-1] Electrochemical study of the function of antibodies immobilized through chemical absorption and covalent binding Sara Nourani, Khadijeh Eskandari, Hedayatolah Ghourchian Tehran University, Tehran, Iran E-mail addresses: [email protected] (S. Nourani), [email protected] (K. Eskandari), [email protected] (H. Ghourchian)
E Poster – [A-10-303-1] Nano-light technology-based probe, a tool for detection of apoptotic cells Mahboobeh Nazaria, Rahman Emamzadehb, Saman Hossinkhania a Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran b Department of Biology, Faculty of Science, University of Isfahan, Tehran, Iran