- Email: [email protected]
Effects of NaSCN and KSCN on fatty acid absorption by E.G.S from small intestinal of sheep
S266
Abstracts
Poster – [A-10-1131-1] Comparing serum Follicle-Stimulating Hormone (FSH) level with vaginal pH in women with menopausal symptoms Zohreh Yousefi, Shohreh Saeed, Fatemeh Vahidroodsari, Sedigheh Ayati Ghaem Hospital, School of Medical Sciences, Mashhad University of Medical Sciences, Mashhad, Iran E-mail addresses: [email protected] (Z. Yousefi), [email protected] (S. Saeed), [email protected] (F. Vahidroodsari), [email protected] (S. Ayati)
Materials and methods: Tubulin was extracted from brain sheep and was conducted by adding 1 mM GTP to tubulin in PEM buffer at 37 ° C for 10–12 min in the presence and absence of cadaverine. Results: The results showed that cadaverine relatively enhanced microtubule polymerization rate at low concentrations (micromolar). But it compeletely inhibited the polymerization with increasing concentration up to millimolars. Moreover, nucleation time of tubuline polymerization increased with increasing concentrations of cadaverine. Conclusion: Therefore, according to our observations, effect of cadaverine on microtubule polymerization showed concentration dependency.
Introduction: Despite the important implication for women's health and reproduction, very few studies have focused on vaginal pH for menopausal diagnosis. Recent studies have suggested vaginal pH as a simple, noninvasive and inexpensive method for this purpose. Objective: The aim of this study is to compare serum FSH level with vaginal PH in menopause. Methods: This is a cross-sectional, descriptive study, conducted on 103 women (aged 31–95 yrs) with menopausal symptoms who were referred to the Menopausal Clinic at Ghaem Hospital during 2006. Vaginal pH was measured using pH meter strips and serum FSH levels were measured using immunoassay methods. The data was analyzed using SPSS software (version 11.5) and results were evaluated statistically by the Chi-square and Kappa tests. p≤0.05 was considered statistically significant. Results: According to this study, in the absence of vaginal infection, the average vaginal pH in these 103 menopausal women was 5.33±0.53. If the menopausal hallmark was considered as vaginal pH>4.5, and serum FSH as ≥20 mIU/ml, then the sensitivity of vaginal pH for menopausal diagnosis was 97%. The mean of FSH levels in this population was 80.79 mIU/ml. Conclusion: Vaginal pH is a simple, accurate, and cost effective tool that can be suggested as a suitable alternative to serum FSH measurement for the diagnosis of menopause.
Keywords: Microtubule, Polymerization, Cadaverine, Nucleation
Keywords: FSH, Vaginal pH, Menopausal symptoms doi:10.1016/j.clinbiochem.2011.08.920
Poster – [A-10-1134-1] Cadaverine effect on microtubule polymerization Tayebe Cheraghi-Shavia, Roya Mahinpurb, Gholamhosein Riazic, Zahra Moosavi-Nejadd a Tehran, Alzahra University, Science Faculty Biology Department, Iran b Faculty of Science, Kashan University, Kashan, Iran c Tehran, Tehran University, Institute of Biochemistry and Biophysics IBB, Iran d Biology Department, Iran E-mail addresses: [email protected] (T. Cheraghi-Shavi), [email protected] (R. Mahinpur), [email protected] (G. Riazi) Introduction: Microtubules are key components of the cytoskeleton of eukaryotic cells and play important roles in many aspects of biological functions including intracellular trafficking of vesicles and organelles, cellular motility and mitotic chromosome segregation. Microtubules are highly dynamic structures whose regulation is crucial for variety of cellular processes. Several molecules have been shown to affect the kinetics of the process. Some kinetically active substances may affect the structural pattern of the polymer. Development of microtubule inhibitors, which interfere with microtubule assembly and disassembly, is useful for the investigation of the biological function of microtubules. In this study, effect of cadaverine as a biological ligand has been investigated on microtubule polymerization.
doi:10.1016/j.clinbiochem.2011.08.921
Poster – [A-10-1135-1] Spectrophotometric investigation of copper (II)–erythromycin interaction Hillsa Bidkhori, Raziyeh Sanavikhoshnood, Zarin Eshaghi Department of Chemistry, Mashhad Branch, Islamic Azad University of Mashhad, Mashhad, Iran E-mail addresses: [email protected] (H. Bidkhori), [email protected] (R. Sanavikhoshnood), [email protected] (Z. Eshaghi) Erythromycin is a macrolid antibiotic consisting of a 14 member ring , a keton group, two glycosidic bonds and dimethylamino group. The drug targets the ribosome and inhibits the protein synthesis of Gram positive bacteria. In this work, we studied the interaction of erythromycin with Cu2+ cation in methanol solvent and determined the formation constant of the possible complex that can be produced by this interaction. Clinical work showed that the concurrent ingestion of antibiotics containing multivalent cations could reduce the bioavailability by up to 90%. Job's method of continuous variation was employed. The interaction of erythromycin with Cu2+ was studied by preparation of a series of methanolic solutions with variation concentraction of ligand and cation. Measurment on complex was done at the wavelength of 340 nm and the influence of pH, time and interferences has been tested. The interaction of erythromycin with Cu2+ has been found to form one complex with metal to ligand composition of 1:2 and the formation constant was obtained to indicate that the complex is stable. Keywords: Spectrophotometry, Erythromycin, Interaction, Cu2+ doi:10.1016/j.clinbiochem.2011.08.922
Poster – [A-10-1157-1] Effects of NaSCN and KSCN on fatty acid absorption by E.G.S from small intestinal of sheep Heshmatallah Oroojia, Mohsen Anib, Seyed Aliasghar Moshtaghib a Najafabad Azad University, Najafabd, Iran b Department of Biochemistry, Isfahan University of Medical Sciences, Isfahan, Iran E-mail addresses: [email protected] (H. Orooji), [email protected] (S.A. Moshtaghi) Introduction: Fatty acids are lipophilic molecules which were believed that they can pass easily across the membrane. But studies revealed that fatty acid transport occurs by special transporters which are present in many tissues. More studies showed that for this transport Na+ is also needed so that in the absence of Na+, fatty acid uptake is
Abstracts
significantly inhibited. To see whether this transport mechanism is affected by inhibitors of active transport, this study was designed and the rate of fatty acid uptake by E.G.S at different conditions was investigated. Method: E.G.S (8–10 cm long) was prepared using the jejunum of sheep and filled with appropriate buffer. Sacs were incubated in a solution containing fatty acids (2 mM) and in a solution containing fatty acids (2 mM) in which Na+ (100 mM) was also present. The transport of fatty acids across the membrane was evaluated by measuring the level of fatty acids inside the sacs. Some procedures were performed except that NaSCN (100 mM) and/or KSCN (100 mM) was also included in the solution. After 20 min of incubation, fatty acids were measured in the buffer inside the E.G.S and compared with controls. Results: Results showed that Na+ is essential for transport of fatty acids across the cell membrane, thus 85% increase was observed when Na+ was present and NaSCN and KSCN increase fatty acid uptake by 650 and 65 percent respectively. Conclusion: Many substances are absorbed across membranes by a co-transport mechanism in which Na+ is essential for the transport. Although most of these transportations are passive Na+ dependent processes but inhibitors of active transport are also effective. The results of the present study showed that although Na+ presence is the essential part of this transport system, but vanadate and ovabain could also interfere with fatty acid transport presumably by inhibition of Na+–K+ ATPases in the membrane. Keywords: NaSCN, KSCN, Absorption, E.G.S doi:10.1016/j.clinbiochem.2011.08.923
Poster – [A-10-1177-2] Studies on the interaction of colchicine and human serum albumin by the measurement of zeta-potential Masoomeh Vahidzadeha, Mohsen Gharanfolib, Jamshid Chamania a Islamic Azad University-Mashhad Branch, Mashhad, Iran b Culture and Science, University E-mail addresses: [email protected] (M. Vahidzadeh), [email protected] (M. Gharanfoli), [email protected] (J. Chamani) Introduction: Human serum albumin (HSA) is the most abundant protein in human blood plasma. It plays an important role in storing and carrying drugs in human blood. Colchicine (COL) is the main alkaloid contained in Colchicum autumnale L. The plant is considered to be very toxic to man and herbivorous animals due to the presence of alkaloids of colchicine type in all of its organs. The interaction of HSA and COL studies by measurement of zeta-potential. Methods: Zeta potential measurement followed for the same sample using Particle size Analyzer apparatus. The potential was calculated from the electrophoretic mobility using assuming the double layer thickness is much less than the particle size. Result and discussion: In the present study, we determined the effect of zeta potential of drug on adsorption of HSA and at low concentration of drug, the zeta potential increase that shows electrostatic interaction predominates HSA and drug. Conclusion: With increasing of COL concentration, the zeta potential values that determine the hydrophobic interaction play an important role in HAS-drug complexes as binary systems. Keywords: HSA, Colchicine, Zeta potential doi:10.1016/j.clinbiochem.2011.08.924
S267
Poster – [A-10-1213-1] Reducing allergenicity of bovine beta lactoglobulin throughout Cu complexes Sajedeh Ebrahim-Damavandia, Divsalar Adelehb, Ali Akbar Sabourya a Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran b Tehran-Karaj, Iran E-mail addresses: [email protected] (S. Ebrahim-Damavandi), [email protected] (A.A. Saboury) Introduction: Bovine milk allergy arises from stable proteins structure. This structure causes gastrointestinal digestion resistance against proteases. Hence intact fragments and protein lead to allergic reaction. Beta lactoglobulin (BLG) is one of the main allergens of bovine milk. Proteolytic role of Cu complexes has been studied previously. Also they may alter protein structure so Cu complexes can assist proteases to digest BLG. Methods: Two new Cu complexes [Cu(dien)OH2](NO3)2 and [Cu (trien)](NO3)2 were designed and characterized. BLG was incubated separately for 30 h with Cu complexes. SDS-PAGE was used to check proteolysis. Extent of hydrolysis was measured by fluorescence methods. Allergenicity of 30 hour incubated protein and protein without Cu complexes was compared by using ELISA. Spectroscopic methods were used to record modification of protein structure. Results: Fluorescence, UV and CD approved Cu complexes bind to the protein, which can modify the protein structure. According to SDS-PAGE results, the protein was cleaved. The allergenicity of the protein treated by Cu complexes showed less allergenicity for the protein incubated with chymotrypsin in the presence of Cu complexes with respect to the absence of Cu complexes. Conclusion: The resistant structure of allergen (BLG) can be changed related to fragmentization of the protein by Cu complexes. So Cu complexes can increase extent of proteolysis by proteases during the digestion process. Keywords: BLG, Cu complex, Allergy doi:10.1016/j.clinbiochem.2011.08.925
Poster – [A-10-1214-1] Spectroscopic evidences for heme degradation during hemoglobin fructation Masume Goodarzia, Ali Akbar Sabourya, Mehran Habibi-Rezaeib, Ali Akbar Moosavi-Mvahedia a IBB, Tehran University, Enghelab Avenue, Tehran, Iran b School of Biology, Tehran University, Enghelab Avenue, Tehran E-mail addresses: [email protected] (M. Goodarzi), [email protected] (A.A. Saboury), [email protected] (M. Habibi-Rezaei), [email protected] (A.A. Moosavi-Mvahedi) Introduction: Nonenzymatic covalent attachment among sugars and protein's amino groups is called glycation, which is made up of several consecutive reactions. This modification occurs at hyperglycemic condition in some places, which may have no regulation for sugar entry. In diabetic patients hemoglobin (Hb) is a suitable target for fructation (glycation with fructose is called fructation) because of noticeable fructose enhancement in Red Blood Cells (RBCs) at hyperglycemic condition. Hemoglobin fructation resulted in its structural and functional modifications. Methods: In this research we focused on heme properties of fructated hemoglobin in invitro physiological-like condition by fluorescence and UV–Vis spectroscopic techniques at different times of incubation. Results: Surprisingly fluorescence results showed the production of heme degradation products with an increasing pattern with respect to the
Abstracts
Poster – [A-10-1131-1] Comparing serum Follicle-Stimulating Hormone (FSH) level with vaginal pH in women with menopausal symptoms Zohreh Yousefi, Shohreh Saeed, Fatemeh Vahidroodsari, Sedigheh Ayati Ghaem Hospital, School of Medical Sciences, Mashhad University of Medical Sciences, Mashhad, Iran E-mail addresses: [email protected] (Z. Yousefi), [email protected] (S. Saeed), [email protected] (F. Vahidroodsari), [email protected] (S. Ayati)
Materials and methods: Tubulin was extracted from brain sheep and was conducted by adding 1 mM GTP to tubulin in PEM buffer at 37 ° C for 10–12 min in the presence and absence of cadaverine. Results: The results showed that cadaverine relatively enhanced microtubule polymerization rate at low concentrations (micromolar). But it compeletely inhibited the polymerization with increasing concentration up to millimolars. Moreover, nucleation time of tubuline polymerization increased with increasing concentrations of cadaverine. Conclusion: Therefore, according to our observations, effect of cadaverine on microtubule polymerization showed concentration dependency.
Introduction: Despite the important implication for women's health and reproduction, very few studies have focused on vaginal pH for menopausal diagnosis. Recent studies have suggested vaginal pH as a simple, noninvasive and inexpensive method for this purpose. Objective: The aim of this study is to compare serum FSH level with vaginal PH in menopause. Methods: This is a cross-sectional, descriptive study, conducted on 103 women (aged 31–95 yrs) with menopausal symptoms who were referred to the Menopausal Clinic at Ghaem Hospital during 2006. Vaginal pH was measured using pH meter strips and serum FSH levels were measured using immunoassay methods. The data was analyzed using SPSS software (version 11.5) and results were evaluated statistically by the Chi-square and Kappa tests. p≤0.05 was considered statistically significant. Results: According to this study, in the absence of vaginal infection, the average vaginal pH in these 103 menopausal women was 5.33±0.53. If the menopausal hallmark was considered as vaginal pH>4.5, and serum FSH as ≥20 mIU/ml, then the sensitivity of vaginal pH for menopausal diagnosis was 97%. The mean of FSH levels in this population was 80.79 mIU/ml. Conclusion: Vaginal pH is a simple, accurate, and cost effective tool that can be suggested as a suitable alternative to serum FSH measurement for the diagnosis of menopause.
Keywords: Microtubule, Polymerization, Cadaverine, Nucleation
Keywords: FSH, Vaginal pH, Menopausal symptoms doi:10.1016/j.clinbiochem.2011.08.920
Poster – [A-10-1134-1] Cadaverine effect on microtubule polymerization Tayebe Cheraghi-Shavia, Roya Mahinpurb, Gholamhosein Riazic, Zahra Moosavi-Nejadd a Tehran, Alzahra University, Science Faculty Biology Department, Iran b Faculty of Science, Kashan University, Kashan, Iran c Tehran, Tehran University, Institute of Biochemistry and Biophysics IBB, Iran d Biology Department, Iran E-mail addresses: [email protected] (T. Cheraghi-Shavi), [email protected] (R. Mahinpur), [email protected] (G. Riazi) Introduction: Microtubules are key components of the cytoskeleton of eukaryotic cells and play important roles in many aspects of biological functions including intracellular trafficking of vesicles and organelles, cellular motility and mitotic chromosome segregation. Microtubules are highly dynamic structures whose regulation is crucial for variety of cellular processes. Several molecules have been shown to affect the kinetics of the process. Some kinetically active substances may affect the structural pattern of the polymer. Development of microtubule inhibitors, which interfere with microtubule assembly and disassembly, is useful for the investigation of the biological function of microtubules. In this study, effect of cadaverine as a biological ligand has been investigated on microtubule polymerization.
doi:10.1016/j.clinbiochem.2011.08.921
Poster – [A-10-1135-1] Spectrophotometric investigation of copper (II)–erythromycin interaction Hillsa Bidkhori, Raziyeh Sanavikhoshnood, Zarin Eshaghi Department of Chemistry, Mashhad Branch, Islamic Azad University of Mashhad, Mashhad, Iran E-mail addresses: [email protected] (H. Bidkhori), [email protected] (R. Sanavikhoshnood), [email protected] (Z. Eshaghi) Erythromycin is a macrolid antibiotic consisting of a 14 member ring , a keton group, two glycosidic bonds and dimethylamino group. The drug targets the ribosome and inhibits the protein synthesis of Gram positive bacteria. In this work, we studied the interaction of erythromycin with Cu2+ cation in methanol solvent and determined the formation constant of the possible complex that can be produced by this interaction. Clinical work showed that the concurrent ingestion of antibiotics containing multivalent cations could reduce the bioavailability by up to 90%. Job's method of continuous variation was employed. The interaction of erythromycin with Cu2+ was studied by preparation of a series of methanolic solutions with variation concentraction of ligand and cation. Measurment on complex was done at the wavelength of 340 nm and the influence of pH, time and interferences has been tested. The interaction of erythromycin with Cu2+ has been found to form one complex with metal to ligand composition of 1:2 and the formation constant was obtained to indicate that the complex is stable. Keywords: Spectrophotometry, Erythromycin, Interaction, Cu2+ doi:10.1016/j.clinbiochem.2011.08.922
Poster – [A-10-1157-1] Effects of NaSCN and KSCN on fatty acid absorption by E.G.S from small intestinal of sheep Heshmatallah Oroojia, Mohsen Anib, Seyed Aliasghar Moshtaghib a Najafabad Azad University, Najafabd, Iran b Department of Biochemistry, Isfahan University of Medical Sciences, Isfahan, Iran E-mail addresses: [email protected] (H. Orooji), [email protected] (S.A. Moshtaghi) Introduction: Fatty acids are lipophilic molecules which were believed that they can pass easily across the membrane. But studies revealed that fatty acid transport occurs by special transporters which are present in many tissues. More studies showed that for this transport Na+ is also needed so that in the absence of Na+, fatty acid uptake is
Abstracts
significantly inhibited. To see whether this transport mechanism is affected by inhibitors of active transport, this study was designed and the rate of fatty acid uptake by E.G.S at different conditions was investigated. Method: E.G.S (8–10 cm long) was prepared using the jejunum of sheep and filled with appropriate buffer. Sacs were incubated in a solution containing fatty acids (2 mM) and in a solution containing fatty acids (2 mM) in which Na+ (100 mM) was also present. The transport of fatty acids across the membrane was evaluated by measuring the level of fatty acids inside the sacs. Some procedures were performed except that NaSCN (100 mM) and/or KSCN (100 mM) was also included in the solution. After 20 min of incubation, fatty acids were measured in the buffer inside the E.G.S and compared with controls. Results: Results showed that Na+ is essential for transport of fatty acids across the cell membrane, thus 85% increase was observed when Na+ was present and NaSCN and KSCN increase fatty acid uptake by 650 and 65 percent respectively. Conclusion: Many substances are absorbed across membranes by a co-transport mechanism in which Na+ is essential for the transport. Although most of these transportations are passive Na+ dependent processes but inhibitors of active transport are also effective. The results of the present study showed that although Na+ presence is the essential part of this transport system, but vanadate and ovabain could also interfere with fatty acid transport presumably by inhibition of Na+–K+ ATPases in the membrane. Keywords: NaSCN, KSCN, Absorption, E.G.S doi:10.1016/j.clinbiochem.2011.08.923
Poster – [A-10-1177-2] Studies on the interaction of colchicine and human serum albumin by the measurement of zeta-potential Masoomeh Vahidzadeha, Mohsen Gharanfolib, Jamshid Chamania a Islamic Azad University-Mashhad Branch, Mashhad, Iran b Culture and Science, University E-mail addresses: [email protected] (M. Vahidzadeh), [email protected] (M. Gharanfoli), [email protected] (J. Chamani) Introduction: Human serum albumin (HSA) is the most abundant protein in human blood plasma. It plays an important role in storing and carrying drugs in human blood. Colchicine (COL) is the main alkaloid contained in Colchicum autumnale L. The plant is considered to be very toxic to man and herbivorous animals due to the presence of alkaloids of colchicine type in all of its organs. The interaction of HSA and COL studies by measurement of zeta-potential. Methods: Zeta potential measurement followed for the same sample using Particle size Analyzer apparatus. The potential was calculated from the electrophoretic mobility using assuming the double layer thickness is much less than the particle size. Result and discussion: In the present study, we determined the effect of zeta potential of drug on adsorption of HSA and at low concentration of drug, the zeta potential increase that shows electrostatic interaction predominates HSA and drug. Conclusion: With increasing of COL concentration, the zeta potential values that determine the hydrophobic interaction play an important role in HAS-drug complexes as binary systems. Keywords: HSA, Colchicine, Zeta potential doi:10.1016/j.clinbiochem.2011.08.924
S267
Poster – [A-10-1213-1] Reducing allergenicity of bovine beta lactoglobulin throughout Cu complexes Sajedeh Ebrahim-Damavandia, Divsalar Adelehb, Ali Akbar Sabourya a Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran b Tehran-Karaj, Iran E-mail addresses: [email protected] (S. Ebrahim-Damavandi), [email protected] (A.A. Saboury) Introduction: Bovine milk allergy arises from stable proteins structure. This structure causes gastrointestinal digestion resistance against proteases. Hence intact fragments and protein lead to allergic reaction. Beta lactoglobulin (BLG) is one of the main allergens of bovine milk. Proteolytic role of Cu complexes has been studied previously. Also they may alter protein structure so Cu complexes can assist proteases to digest BLG. Methods: Two new Cu complexes [Cu(dien)OH2](NO3)2 and [Cu (trien)](NO3)2 were designed and characterized. BLG was incubated separately for 30 h with Cu complexes. SDS-PAGE was used to check proteolysis. Extent of hydrolysis was measured by fluorescence methods. Allergenicity of 30 hour incubated protein and protein without Cu complexes was compared by using ELISA. Spectroscopic methods were used to record modification of protein structure. Results: Fluorescence, UV and CD approved Cu complexes bind to the protein, which can modify the protein structure. According to SDS-PAGE results, the protein was cleaved. The allergenicity of the protein treated by Cu complexes showed less allergenicity for the protein incubated with chymotrypsin in the presence of Cu complexes with respect to the absence of Cu complexes. Conclusion: The resistant structure of allergen (BLG) can be changed related to fragmentization of the protein by Cu complexes. So Cu complexes can increase extent of proteolysis by proteases during the digestion process. Keywords: BLG, Cu complex, Allergy doi:10.1016/j.clinbiochem.2011.08.925
Poster – [A-10-1214-1] Spectroscopic evidences for heme degradation during hemoglobin fructation Masume Goodarzia, Ali Akbar Sabourya, Mehran Habibi-Rezaeib, Ali Akbar Moosavi-Mvahedia a IBB, Tehran University, Enghelab Avenue, Tehran, Iran b School of Biology, Tehran University, Enghelab Avenue, Tehran E-mail addresses: [email protected] (M. Goodarzi), [email protected] (A.A. Saboury), [email protected] (M. Habibi-Rezaei), [email protected] (A.A. Moosavi-Mvahedi) Introduction: Nonenzymatic covalent attachment among sugars and protein's amino groups is called glycation, which is made up of several consecutive reactions. This modification occurs at hyperglycemic condition in some places, which may have no regulation for sugar entry. In diabetic patients hemoglobin (Hb) is a suitable target for fructation (glycation with fructose is called fructation) because of noticeable fructose enhancement in Red Blood Cells (RBCs) at hyperglycemic condition. Hemoglobin fructation resulted in its structural and functional modifications. Methods: In this research we focused on heme properties of fructated hemoglobin in invitro physiological-like condition by fluorescence and UV–Vis spectroscopic techniques at different times of incubation. Results: Surprisingly fluorescence results showed the production of heme degradation products with an increasing pattern with respect to the