- Email: [email protected]
Transfection of neural stem cells by BDNF gene transfered using nanocarreir system
S222
Abstracts
a
Isfahan University, Isfahan, Iran Tarbiyat Modares University, Iran E-mail addresses: [email protected] (S. Minaeian), [email protected] (F. Rahbarizadeh), [email protected] (S.H. Zarkesh Esfahani)
b
Introduction: Human papilloma virus (HPV) is the main cause of cervical cancer. Cervical cancer is the second commonest cancer among women. Regular cervical screening is the best way to detect changes to the cervix early. But many studies have shown that cytology has a limited sensivity for detecting cervical cancer. Nanobody is functional antibody without light chains and its antigen binding site compromised in single domain fragment. Their diameter is 2.8 nm and their length is about 4.4 nm. These tiny antibodies are very stable, highly soluble and target to the tumor cells very specifically. The ability to conjugate nanobody to another protein or molecule is wonderful for many applications in therapeutics and diagnosis. Methods: In this study we enriched the nanobody library against L1 protein of HPV. After that specific nanobodies against HPV L1 protein were selected. They were expressed in Roseta gami. The nanobodies were purified by immobilized metal affinity chromography. Nanobodies were conjugated to horseradish peroxidase (HRP) and the checker board assay of this product was measured by ELISA. Results: The results demonstrated that purification and conjugation process were performed effectively and perfectly. Conclusion: This specific and sensitive conjugate is cost effective product and can be used in early diagnosis of cervical cancer. Nanobody can be easily formatted in to multifunctional protein so it can be a powerful tool for diagnosis and treatment of many diseases such as cancer. Keywords: Nanobody, HPV, Cervical cancer, HRP
doi:10.1016/j.clinbiochem.2011.08.980
Poster – [A-10-1151-1] Simulation the protein channels of biological membranes using nanoscience to study and treat disorders of ionic channels Reza Rasoolzzadeh, Majid Monajjemi, Maryam Mousavi, Maryam Falahati Tehran, Iran E-mail addresses: [email protected] (R. Rasoolzzadeh), [email protected] (M. Monajjemi), [email protected] (M. Mousavi), [email protected] (M. Falahati) To reconstruct membranos proteins, we used simulated nanotubes in order to transfer ions across the membrane and transfer of ions was done successfully. In this study the more the potential energy increases the more the conductivity of nanochannels decreases and we chose the least energy among nanotube and amino acid complexes. And also the more energy we use, the more conductivity we will have; therefore we choose the complex which conducts the most current. This way we can simulate the channels which have hereditary defects and are not efficient and observe the fundamental cure of the diseases. Keywords: Membranos proteins, Nanotube, Membrane channels, Amino acids
doi:10.1016/j.clinbiochem.2011.08.981
Poster — [A-10-1215-1] A new and highly sensitive nanobiosensor for detection of Helicobacter based on fluorescence resonance energy transfer Shamsipur Mojtabaa, Shanehsaz Maryamb, Mohsenifar Afshinb, Hasannia Sadeghd, Pirooznia Nazaninc, Shahrashoob Mahsad a Razi University, Iran b Tarbiat Modares University, Tehran, Iran c Guilan University, Iran d Payam Noor University, Iran E-mail address: [email protected] (S. Maryam) Introduction: A nanobiosensor based on fluorescence resonance energy transfer (FRET) for detection of Helicobacter using two specified oligonucleotides conjugated with quantum dot and tamra was fabricated. Luminescent semiconductor quantum dots have been intensely studied due to their unique optical properties. In particular, semiconductor QDs are very attractive as biological labels because of their small size, emission tunability, superior photostability and longer photoluminescence (PL) decay times in comparison to conventional organic dyes. Methods: The water-soluble CdSe/ZnS QDs were prepared. Subsequently, two specific oligonucleotides for Helicobacter were attached to the QDs and tamra to form functional QD- and tamraoligonucleotide conjugates. Result: Along with the addition of DNA targets to a solution containing QD- and tamra-oligonucleotide conjugates, hybridization was effectively formed. The resulting assembly brings the tamra fluorophore as an acceptor and the QDs as a donor into the proximity of each other, leading to fluorescence emission transfer from the acceptor by means of FRET phenomenon on illumination of the donor. In the absence of target DNA, the detection probes were not ligated and no tamra emission was produced due to the lack of FRET. Conclusion: The feasibility of the proposed method was also demonstrated in the detection of synthetic 210-mer nucleotide derived from Helicobacter with a sensitivity of 4.0 nM. So here, we conclude that the proposed homogeneously DNA detection method which is simple, rapid and efficient due to the elimination of the washing and separation steps, may be useful in fabrication of a high sensitive nanobiosensor for detection of Helicobacter pylori. Keywords: Helicobacter pylori, Biosensor, CdTe quantum dot-conjugate, FRET doi:10.1016/j.clinbiochem.2011.08.982
Poster – [A-10-1230-1] Transfection of neural stem cells by BDNF gene transfered using nanocarreir system Salim Albukhaty, Hosein Naderimanesh, Taki Tiraihi Shafa Neuroscience Research Center, Medical Center Khatam Al-Anbia Hospital, Dept. of biological science, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran E-mail address: [email protected] (S. Albukhaty) Stem cell and gene therapy can be defined as a part of a group of new techniques, These new techniques are being applied experimentally to a wide range of human disorders, including many types of cancer, neurological diseases such as Parkinson's disease, spinal cord injuries, and diabetes. stem cells transplantation has been shown to promote regeneration and neuro protection in central nervous system (CNS) disorders and spinal cord injuries. nano particles have been used to bring stem cells to sites of neural injury in a new method designed to increase the capacity of cells to repair damaged tissue. BDNF induces neuronal survival and tissue repair after
Abstracts
neurological diseases and (SCI). A continuous BDNF supply is believed to gain greater efficacy in the neural restoration of the injured spinal cord. Accordingly, nano vehicle formulation for their efficient delivery and sustained release in injured spinal cord was examined. According to the existence of (BSCB), many therapeutic macromolecular agents, such as drugs, protein and gene, cannot pass through this barrier to reach the site of injury, all of which restricts the treatment of spinal cord injuries (SCI). In this study, we transfected neural stem cells with the BDNF gene using a nano carreir system and Optimized transfected NSC with BDNF using nanoparticle results in increase in expression of BDNF. Keywords: Transfection, Stem cell, BDNF, Liposome, Nanocarrier system doi:10.1016/j.clinbiochem.2011.08.983
Poster – [A-10-1245-1] Bacterial effect of silver nanoparticles against multidrug-resistant bacteria Monir Doudia, Marziyeh Karamib, Nour Amirmozafaric a Department of Microbiology, Falavarjan Brunch, Islamic Azad University, Falavarjan, Iran b Young Researchers Club, Falavarjan Branch, Islamic Azad University, Iran c Iran University of Medical Sciences, School of Medicine, Tehran, Iran E-mail addresses: [email protected] (M. Doudi), [email protected] (M. Karami), [email protected] (N. Amirmozafari) Introduction: Infections caused by drug-resistant microorganisms result in significant in creases in mortality, morbidity, and cost related to prolonged treatments. The antibacterial activity of silver nanoparticles against some drug-resistant bacteria has been established, but further investigation is needed to determine whether these particles could be an option for the treatment and prevention of drug-resistant microbial infections. Materials and Methods: Hence, we challenged different drugresistant pathogens of clinical importance multidrug-resistant Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus and Streptococcus pyogenes. By using a suspension of silver nanoparticles (25, 50, 100, 200, 250 ppm) in 3 hospital of Esfahan city (Gharazi, Sina and Alzahra). All of the isolated was identified using conventional bacteriologic methods and were tested for sensitivity through a Kirby-Bauer test. The data presented here are novel because they prove that silver nanoparticles are effective bactericidal agents. Results: Regardless of the drug-resistance bacteria and show the importance of silver nanoparticles in the 250 ppm. Conclusion: Silver nanoparticles can be recommended as an effective broad-Spectrum bactericidal agent. Keywords: Nanosilver, Resistant multi drug, Bacterial gram negative and gram positive doi:10.1016/j.clinbiochem.2011.08.984
Poster — [A-10-1246-1] Quantum dots in semiconductor chemical sensors and biosensors Zakavati Royaa, Bayat Mansourb, Mohsenifar Afshinc, Hashemi Hezaveh Gamald a Azad University of Ahwaz, Iran b ﻭﺍﺣﺪﻋﻠﻮﻡ ﲢﻘﯿﻘﺎﺕ ﲥﺮﺍﻥ, Iran c ﺩﺍﻧﺸﮕﺎﻩ ﺗﺮﺑﯿﺖ ﻣﺪﺭﺱ ﲥﺮﺍﻥ, Iran d ﺩﺍﻧﺸﮑﺪﻩ ﲠﺪﺍﺷﺖ ﲥﺮﺍﻥ, Iran E-mail address: [email protected] (Z. Roya)
S223
Show interesting phenomena, such as dependent and show interesting emission wavelength, narrow emission peak and broad excitation rang. The confinement can be received the semiconductor in very small size, and several hundred to thousands quantum dots are semiconductor nano-crystals, which have unique properties and of atoms per particle. Electrons in QDs acquire energy and excitions. An excition is a bound state of an electron called a hole. After relaxation from excitonic excited state of lower energy state and hole recombine. This process emitting a photon recombination of electrons and hole and fluorescent emission is called photoluminescence. The number of photons can be measured as a function of energy, which gives us the PL spectrum. Different from many organic dyes QDs excited by many light sources within a large wavelength range. These properties of QDs are using fluorophores for biological imaging and designing specific nanobiosensors for diagnostic of toxins. Methodology: Containing of synthesis nano-particle of QDs and preparation of aflatoxinB1 solution that was prepared in phosphate buffer and covalent immobilization of anti-AFB1 and aflatoxin conjugate signal, which can be obtained in the signal-making that measured by spectrofluorimetry. Conclusion and discussion: In this study, a biological nanobiosensor aflatoxin is designed to identify which biological sensor being sensitive to traditional methods like ELISA or similar diagnosis will aflatoxin. Antibodies bind to nano-particles of quantum dots aflatoxin connections and maintains the use of antibody-stabilized quantum dots attached to albumin aflatoxin rhodamin able to create a signal and there spectrofluorimetry system there aflatoxin lead to changes in environmental signals be obtained. Keywords: Quantum dots, Biosensors, AflatoxinB1 doi:10.1016/j.clinbiochem.2011.08.985
Poster – [A-10-1248-2] Telomerase activity in patients with leukemia Sadat Khoramgah Maryam Medical University Of Tabriz, Tabriz,Iran E-mail address: [email protected] Introduction: Telomerase is an enzyme that adds DNA sequence repeats ("TTAGGG" in all vertebrates) to the 3' end of DNA strands in the telomere regions, which are found at the ends of eukaryotic chromosomes. This region of repeated nucleotide called telomeres contains non-coding DNA material and prevents constant loss of important DNA from chromosome ends. As a result, every time the chromosome is copied only 100–200 nucleotides are lost, which causes no damage to the organism's DNA. Telomerase is a reverse transcriptase that carries its own RNA molecule, which is used as a template when it elongates telomeres, which are shortened after each replication cycle.TERT is a reverse transcriptase, which is a class of enzyme that creates singlestranded DNA using single-stranded RNA as a template. Method: Samples of bone marrow (BM) and peripheral blood (PB) of patients who previously were not under any treatment, were prepared. PCR reactions for TRAP according to the amount of cell extract major reaction mixture (Master mix) were done. Finally, the reaction products obtained by PCR and ELISA techniques, using probes specific telomere components were evaluated. The technique of PCR products with native-PAGE and then stained with silver nitrate were. Results: Thetelomerase activity is increased in Leukemia patients in comparsion to control subject. Conclusion: significant increased telomerase activity can be used as atumor marker for early diagnosis of Leukemia. Keywords: Telomeras activity, Lukemia doi:10.1016/j.clinbiochem.2011.08.986
Abstracts
a
Isfahan University, Isfahan, Iran Tarbiyat Modares University, Iran E-mail addresses: [email protected] (S. Minaeian), [email protected] (F. Rahbarizadeh), [email protected] (S.H. Zarkesh Esfahani)
b
Introduction: Human papilloma virus (HPV) is the main cause of cervical cancer. Cervical cancer is the second commonest cancer among women. Regular cervical screening is the best way to detect changes to the cervix early. But many studies have shown that cytology has a limited sensivity for detecting cervical cancer. Nanobody is functional antibody without light chains and its antigen binding site compromised in single domain fragment. Their diameter is 2.8 nm and their length is about 4.4 nm. These tiny antibodies are very stable, highly soluble and target to the tumor cells very specifically. The ability to conjugate nanobody to another protein or molecule is wonderful for many applications in therapeutics and diagnosis. Methods: In this study we enriched the nanobody library against L1 protein of HPV. After that specific nanobodies against HPV L1 protein were selected. They were expressed in Roseta gami. The nanobodies were purified by immobilized metal affinity chromography. Nanobodies were conjugated to horseradish peroxidase (HRP) and the checker board assay of this product was measured by ELISA. Results: The results demonstrated that purification and conjugation process were performed effectively and perfectly. Conclusion: This specific and sensitive conjugate is cost effective product and can be used in early diagnosis of cervical cancer. Nanobody can be easily formatted in to multifunctional protein so it can be a powerful tool for diagnosis and treatment of many diseases such as cancer. Keywords: Nanobody, HPV, Cervical cancer, HRP
doi:10.1016/j.clinbiochem.2011.08.980
Poster – [A-10-1151-1] Simulation the protein channels of biological membranes using nanoscience to study and treat disorders of ionic channels Reza Rasoolzzadeh, Majid Monajjemi, Maryam Mousavi, Maryam Falahati Tehran, Iran E-mail addresses: [email protected] (R. Rasoolzzadeh), [email protected] (M. Monajjemi), [email protected] (M. Mousavi), [email protected] (M. Falahati) To reconstruct membranos proteins, we used simulated nanotubes in order to transfer ions across the membrane and transfer of ions was done successfully. In this study the more the potential energy increases the more the conductivity of nanochannels decreases and we chose the least energy among nanotube and amino acid complexes. And also the more energy we use, the more conductivity we will have; therefore we choose the complex which conducts the most current. This way we can simulate the channels which have hereditary defects and are not efficient and observe the fundamental cure of the diseases. Keywords: Membranos proteins, Nanotube, Membrane channels, Amino acids
doi:10.1016/j.clinbiochem.2011.08.981
Poster — [A-10-1215-1] A new and highly sensitive nanobiosensor for detection of Helicobacter based on fluorescence resonance energy transfer Shamsipur Mojtabaa, Shanehsaz Maryamb, Mohsenifar Afshinb, Hasannia Sadeghd, Pirooznia Nazaninc, Shahrashoob Mahsad a Razi University, Iran b Tarbiat Modares University, Tehran, Iran c Guilan University, Iran d Payam Noor University, Iran E-mail address: [email protected] (S. Maryam) Introduction: A nanobiosensor based on fluorescence resonance energy transfer (FRET) for detection of Helicobacter using two specified oligonucleotides conjugated with quantum dot and tamra was fabricated. Luminescent semiconductor quantum dots have been intensely studied due to their unique optical properties. In particular, semiconductor QDs are very attractive as biological labels because of their small size, emission tunability, superior photostability and longer photoluminescence (PL) decay times in comparison to conventional organic dyes. Methods: The water-soluble CdSe/ZnS QDs were prepared. Subsequently, two specific oligonucleotides for Helicobacter were attached to the QDs and tamra to form functional QD- and tamraoligonucleotide conjugates. Result: Along with the addition of DNA targets to a solution containing QD- and tamra-oligonucleotide conjugates, hybridization was effectively formed. The resulting assembly brings the tamra fluorophore as an acceptor and the QDs as a donor into the proximity of each other, leading to fluorescence emission transfer from the acceptor by means of FRET phenomenon on illumination of the donor. In the absence of target DNA, the detection probes were not ligated and no tamra emission was produced due to the lack of FRET. Conclusion: The feasibility of the proposed method was also demonstrated in the detection of synthetic 210-mer nucleotide derived from Helicobacter with a sensitivity of 4.0 nM. So here, we conclude that the proposed homogeneously DNA detection method which is simple, rapid and efficient due to the elimination of the washing and separation steps, may be useful in fabrication of a high sensitive nanobiosensor for detection of Helicobacter pylori. Keywords: Helicobacter pylori, Biosensor, CdTe quantum dot-conjugate, FRET doi:10.1016/j.clinbiochem.2011.08.982
Poster – [A-10-1230-1] Transfection of neural stem cells by BDNF gene transfered using nanocarreir system Salim Albukhaty, Hosein Naderimanesh, Taki Tiraihi Shafa Neuroscience Research Center, Medical Center Khatam Al-Anbia Hospital, Dept. of biological science, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran E-mail address: [email protected] (S. Albukhaty) Stem cell and gene therapy can be defined as a part of a group of new techniques, These new techniques are being applied experimentally to a wide range of human disorders, including many types of cancer, neurological diseases such as Parkinson's disease, spinal cord injuries, and diabetes. stem cells transplantation has been shown to promote regeneration and neuro protection in central nervous system (CNS) disorders and spinal cord injuries. nano particles have been used to bring stem cells to sites of neural injury in a new method designed to increase the capacity of cells to repair damaged tissue. BDNF induces neuronal survival and tissue repair after
Abstracts
neurological diseases and (SCI). A continuous BDNF supply is believed to gain greater efficacy in the neural restoration of the injured spinal cord. Accordingly, nano vehicle formulation for their efficient delivery and sustained release in injured spinal cord was examined. According to the existence of (BSCB), many therapeutic macromolecular agents, such as drugs, protein and gene, cannot pass through this barrier to reach the site of injury, all of which restricts the treatment of spinal cord injuries (SCI). In this study, we transfected neural stem cells with the BDNF gene using a nano carreir system and Optimized transfected NSC with BDNF using nanoparticle results in increase in expression of BDNF. Keywords: Transfection, Stem cell, BDNF, Liposome, Nanocarrier system doi:10.1016/j.clinbiochem.2011.08.983
Poster – [A-10-1245-1] Bacterial effect of silver nanoparticles against multidrug-resistant bacteria Monir Doudia, Marziyeh Karamib, Nour Amirmozafaric a Department of Microbiology, Falavarjan Brunch, Islamic Azad University, Falavarjan, Iran b Young Researchers Club, Falavarjan Branch, Islamic Azad University, Iran c Iran University of Medical Sciences, School of Medicine, Tehran, Iran E-mail addresses: [email protected] (M. Doudi), [email protected] (M. Karami), [email protected] (N. Amirmozafari) Introduction: Infections caused by drug-resistant microorganisms result in significant in creases in mortality, morbidity, and cost related to prolonged treatments. The antibacterial activity of silver nanoparticles against some drug-resistant bacteria has been established, but further investigation is needed to determine whether these particles could be an option for the treatment and prevention of drug-resistant microbial infections. Materials and Methods: Hence, we challenged different drugresistant pathogens of clinical importance multidrug-resistant Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus and Streptococcus pyogenes. By using a suspension of silver nanoparticles (25, 50, 100, 200, 250 ppm) in 3 hospital of Esfahan city (Gharazi, Sina and Alzahra). All of the isolated was identified using conventional bacteriologic methods and were tested for sensitivity through a Kirby-Bauer test. The data presented here are novel because they prove that silver nanoparticles are effective bactericidal agents. Results: Regardless of the drug-resistance bacteria and show the importance of silver nanoparticles in the 250 ppm. Conclusion: Silver nanoparticles can be recommended as an effective broad-Spectrum bactericidal agent. Keywords: Nanosilver, Resistant multi drug, Bacterial gram negative and gram positive doi:10.1016/j.clinbiochem.2011.08.984
Poster — [A-10-1246-1] Quantum dots in semiconductor chemical sensors and biosensors Zakavati Royaa, Bayat Mansourb, Mohsenifar Afshinc, Hashemi Hezaveh Gamald a Azad University of Ahwaz, Iran b ﻭﺍﺣﺪﻋﻠﻮﻡ ﲢﻘﯿﻘﺎﺕ ﲥﺮﺍﻥ, Iran c ﺩﺍﻧﺸﮕﺎﻩ ﺗﺮﺑﯿﺖ ﻣﺪﺭﺱ ﲥﺮﺍﻥ, Iran d ﺩﺍﻧﺸﮑﺪﻩ ﲠﺪﺍﺷﺖ ﲥﺮﺍﻥ, Iran E-mail address: [email protected] (Z. Roya)
S223
Show interesting phenomena, such as dependent and show interesting emission wavelength, narrow emission peak and broad excitation rang. The confinement can be received the semiconductor in very small size, and several hundred to thousands quantum dots are semiconductor nano-crystals, which have unique properties and of atoms per particle. Electrons in QDs acquire energy and excitions. An excition is a bound state of an electron called a hole. After relaxation from excitonic excited state of lower energy state and hole recombine. This process emitting a photon recombination of electrons and hole and fluorescent emission is called photoluminescence. The number of photons can be measured as a function of energy, which gives us the PL spectrum. Different from many organic dyes QDs excited by many light sources within a large wavelength range. These properties of QDs are using fluorophores for biological imaging and designing specific nanobiosensors for diagnostic of toxins. Methodology: Containing of synthesis nano-particle of QDs and preparation of aflatoxinB1 solution that was prepared in phosphate buffer and covalent immobilization of anti-AFB1 and aflatoxin conjugate signal, which can be obtained in the signal-making that measured by spectrofluorimetry. Conclusion and discussion: In this study, a biological nanobiosensor aflatoxin is designed to identify which biological sensor being sensitive to traditional methods like ELISA or similar diagnosis will aflatoxin. Antibodies bind to nano-particles of quantum dots aflatoxin connections and maintains the use of antibody-stabilized quantum dots attached to albumin aflatoxin rhodamin able to create a signal and there spectrofluorimetry system there aflatoxin lead to changes in environmental signals be obtained. Keywords: Quantum dots, Biosensors, AflatoxinB1 doi:10.1016/j.clinbiochem.2011.08.985
Poster – [A-10-1248-2] Telomerase activity in patients with leukemia Sadat Khoramgah Maryam Medical University Of Tabriz, Tabriz,Iran E-mail address: [email protected] Introduction: Telomerase is an enzyme that adds DNA sequence repeats ("TTAGGG" in all vertebrates) to the 3' end of DNA strands in the telomere regions, which are found at the ends of eukaryotic chromosomes. This region of repeated nucleotide called telomeres contains non-coding DNA material and prevents constant loss of important DNA from chromosome ends. As a result, every time the chromosome is copied only 100–200 nucleotides are lost, which causes no damage to the organism's DNA. Telomerase is a reverse transcriptase that carries its own RNA molecule, which is used as a template when it elongates telomeres, which are shortened after each replication cycle.TERT is a reverse transcriptase, which is a class of enzyme that creates singlestranded DNA using single-stranded RNA as a template. Method: Samples of bone marrow (BM) and peripheral blood (PB) of patients who previously were not under any treatment, were prepared. PCR reactions for TRAP according to the amount of cell extract major reaction mixture (Master mix) were done. Finally, the reaction products obtained by PCR and ELISA techniques, using probes specific telomere components were evaluated. The technique of PCR products with native-PAGE and then stained with silver nitrate were. Results: Thetelomerase activity is increased in Leukemia patients in comparsion to control subject. Conclusion: significant increased telomerase activity can be used as atumor marker for early diagnosis of Leukemia. Keywords: Telomeras activity, Lukemia doi:10.1016/j.clinbiochem.2011.08.986