- Email: [email protected]
Recombinant production of anti-HIV protein, Griffithsin, in E. coli
Abstracts
Poster – [A-10-1138-1] The acetylcholine esterase activity inhibition as a possible mechanism for beneficial effects of Nigella sativa on memory Mohamadpoor Toktam, Hosseini Mahmood, Karami Reza, Sookhtanloo Mohammad, Rajaei Ziba Emam Reza Hospital, Iran E-mail address: [email protected] (M. Toktam) The acetylcholine esterase activity inhibition as a possible mechanism for beneficial effects of Nigella sativa on memory and neuroprotective, anti-oxidant, anti-convulsant and analgesic effects of Nigella sativa (NS) has been previously reported. In the present study the effect of Nigella sativa memory and acetylcholine esterase(ACE) activity in scopolamine-induced memory impairment model was evaluated. Forty male Wistar rats were divided into five groups: group 1 (Control) received saline instead of both NS extract and scopolamine. The animals of group 2 (Scopolamine; Sco) were treated with saline instead of NS extract but were injected by scopolamine before retention test. Groups 3 (Sco+ NS 200) and 4 (Sco + NS 400) were daily treated by 200 and 400 mg/kg of NS respectively, for 3 weeks and finally injected with scopolamine before retention test. Passive avoidance test was performed for all groups. The brains of the animals were removed to determine the ACE activity. Before receiving shock, there was no significant difference in time latency to enter the dark compartment between groups. The time latency to enter the dark compartment in Scopolamine group was lower than of Control group (P< 0.01). The time latency in Sco + NS 400 group was higher than Scopolamine group (P < 0.01). AChE activity in the both hippocampus and cortex tissues of Scopolamine group was significantly higher than Control group (P < 0.001) while, AChE activity in Sco + NS 400 group was lower than Scopolamine group (P< 0.001). Following scopolamine administration, total SH groups decreased (P < 0.01) and MDA concentrations were increased (P < 0.001) in comparison with Control group. The extracts' pretreatment caused a significant elevation in total thiol groups (P < 0.01). Cortical MDA concentration in extract treated groups were lower than scopolamine group (P < 0.001). The results of present study showed hydro-acoholic extract of Nigella sativa prevents sciopolamine-induced deficit memory in rats. Our result supports the traditional belief about the beneficial effect of Nigella sativa in the nervous system. Moreover for more clarification of this effect further investigations using other methods are recommended in future studies. Keywords: Memory, Scopolamine, Nigella sativa, AChE, Thiol groups, MDA assay doi:10.1016/j.clinbiochem.2011.08.872
Poster — [A-10-1144-1] Chemical constituents and antibacterial activity of essential oil of Anvillea garcini Attarpour yazdi Mohammad Mehdi P.O. Box 18155/159, Tehran, Iran E-mail address: [email protected] Introduction: Anvillea garcini grows wildly in the southern region of Iran. The purpose of this study was to investigate chemical constituents of the essential oil from leaves and flowers of A. garcini and their evaluation of antibacterial activity against Streptococcus pyogenes, Staphylococcus saprophyticus, Enterobacter aerogenes and Bacillus anthracis. Methods: Leaves and flowers of the plant were collected from Bandar-Abbas and the essential oil was extracted by hydro-distillation method and the chemical constituents were investigated by GC/MS method. Antibacterial activity was evaluated by disk diffusion and agar dilution methods for determining of MIC.
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Results: A. garcini leaves and flowers yielded 0.3% of essential oil on hydro-distillation that the major chemical constituents of the leaf oil were α-pinene (15.9%) and sabinene (9.7%) and the flower oil was bornyl acetate (38.8%) and camphene (9.8%). Results from the antibacterial tests demonstrated that the A. garcini leaf and flower oil had an excellent inhibitory effect against the 3 of 4 tested bacteria. The MIC of the leaf and flower oils against the Streptococcus pyogenes, Staphylococcus saprophyticus, Bacillus anthracis and Enterobacter aerogenes were 0.0, 5.0, 5.0 and 10 μg/ml, respectively. Conclusion: This study demonstrated that essential oil of A. garcini leaf and flower and its constituents have excellent antibacterial activity and are beneficial to human health. They have the potential to be used for medical purposes and to be utilized as antibacterial additives in making paper products. However, we need more investigation in vitro and in vivo. Keywords: Anvillea garcini, Streptococcus pyogenes, Staphylococcus saprophyticus, Enterobacter aerogenes, Bacillus anthracis doi:10.1016/j.clinbiochem.2011.08.873
Poster — [A-10-1154-2] Recombinant production of anti-HIV protein, Griffithsin, in E. coli Afshar Shandiz Mohammada, Vafaee Yavarb, Babalar Mesbahb, Alizadeh Houshanga a Agronomy Dep. of Agricultural Biotechnology, Agricultural College, Tehran University, Karaj, Iran b Dept. of Horticultural Science, Agricultural College, Tehran University, Karaj, Iran E-mail addresses: [email protected] (A.S. Mohammad), [email protected] (V. Yavar), [email protected] (B. Mesbah), [email protected] (A. Houshang) Over the past 27 years, nearly 25 million people have died from AIDS. Available strategy such as highly active antiretroviral therapy (HAART) can prevent HIV transmission from parents to children but fail to treat this disease. The red alga Griffithia sp. has a novel antiHIV protein which is called Griffithsin (GRFT). The potent anti-viral activity of GRFT against both laboratory and primary isolates of HIV at picomolar concentrations makes this protein an attractive candidate microbiocide to prevent sexual transmission of HIV. At the first step, we optimized codon usage and designed hexa-histidine tagged GRFT (His-GRFT) sequence for expression in Escherichia coli. Then synthesized GRFT sequence was inserted in expressional vector pET28a (+) and transformed to E. coli BL21 strain. Finally, the recombinant production of GRFT was confirmed by SDS-PAGE. The expression system described in this work can provide a basis for the mass production of GRFT to allow further studies of the protein and investigation of therapeutic and preventive strategies against HIV. Keywords: Griffithsin, Lectin, Anti-HIV, His tag, E. coli doi:10.1016/j.clinbiochem.2011.08.874
Poster — [A-10-1156-1] The effect of combination of aerobic training and oral administration of hydro extract of saffron (Crocus sativus) on serum lipid profile in streptozotosin induced diabetic rats Roya Irajia, Mohamad Ali Azarbayjanib, Mohamad Javad Rasaeec, Peeri Maghsooda, Asghar Nikpoora, Ayati Esfandyara a Ghods sq, Iranzamin St, Islamic Azad University, Central Tehran Branch Faculty of Aerobic Education, Tehran, Iran
Poster – [A-10-1138-1] The acetylcholine esterase activity inhibition as a possible mechanism for beneficial effects of Nigella sativa on memory Mohamadpoor Toktam, Hosseini Mahmood, Karami Reza, Sookhtanloo Mohammad, Rajaei Ziba Emam Reza Hospital, Iran E-mail address: [email protected] (M. Toktam) The acetylcholine esterase activity inhibition as a possible mechanism for beneficial effects of Nigella sativa on memory and neuroprotective, anti-oxidant, anti-convulsant and analgesic effects of Nigella sativa (NS) has been previously reported. In the present study the effect of Nigella sativa memory and acetylcholine esterase(ACE) activity in scopolamine-induced memory impairment model was evaluated. Forty male Wistar rats were divided into five groups: group 1 (Control) received saline instead of both NS extract and scopolamine. The animals of group 2 (Scopolamine; Sco) were treated with saline instead of NS extract but were injected by scopolamine before retention test. Groups 3 (Sco+ NS 200) and 4 (Sco + NS 400) were daily treated by 200 and 400 mg/kg of NS respectively, for 3 weeks and finally injected with scopolamine before retention test. Passive avoidance test was performed for all groups. The brains of the animals were removed to determine the ACE activity. Before receiving shock, there was no significant difference in time latency to enter the dark compartment between groups. The time latency to enter the dark compartment in Scopolamine group was lower than of Control group (P< 0.01). The time latency in Sco + NS 400 group was higher than Scopolamine group (P < 0.01). AChE activity in the both hippocampus and cortex tissues of Scopolamine group was significantly higher than Control group (P < 0.001) while, AChE activity in Sco + NS 400 group was lower than Scopolamine group (P< 0.001). Following scopolamine administration, total SH groups decreased (P < 0.01) and MDA concentrations were increased (P < 0.001) in comparison with Control group. The extracts' pretreatment caused a significant elevation in total thiol groups (P < 0.01). Cortical MDA concentration in extract treated groups were lower than scopolamine group (P < 0.001). The results of present study showed hydro-acoholic extract of Nigella sativa prevents sciopolamine-induced deficit memory in rats. Our result supports the traditional belief about the beneficial effect of Nigella sativa in the nervous system. Moreover for more clarification of this effect further investigations using other methods are recommended in future studies. Keywords: Memory, Scopolamine, Nigella sativa, AChE, Thiol groups, MDA assay doi:10.1016/j.clinbiochem.2011.08.872
Poster — [A-10-1144-1] Chemical constituents and antibacterial activity of essential oil of Anvillea garcini Attarpour yazdi Mohammad Mehdi P.O. Box 18155/159, Tehran, Iran E-mail address: [email protected] Introduction: Anvillea garcini grows wildly in the southern region of Iran. The purpose of this study was to investigate chemical constituents of the essential oil from leaves and flowers of A. garcini and their evaluation of antibacterial activity against Streptococcus pyogenes, Staphylococcus saprophyticus, Enterobacter aerogenes and Bacillus anthracis. Methods: Leaves and flowers of the plant were collected from Bandar-Abbas and the essential oil was extracted by hydro-distillation method and the chemical constituents were investigated by GC/MS method. Antibacterial activity was evaluated by disk diffusion and agar dilution methods for determining of MIC.
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Results: A. garcini leaves and flowers yielded 0.3% of essential oil on hydro-distillation that the major chemical constituents of the leaf oil were α-pinene (15.9%) and sabinene (9.7%) and the flower oil was bornyl acetate (38.8%) and camphene (9.8%). Results from the antibacterial tests demonstrated that the A. garcini leaf and flower oil had an excellent inhibitory effect against the 3 of 4 tested bacteria. The MIC of the leaf and flower oils against the Streptococcus pyogenes, Staphylococcus saprophyticus, Bacillus anthracis and Enterobacter aerogenes were 0.0, 5.0, 5.0 and 10 μg/ml, respectively. Conclusion: This study demonstrated that essential oil of A. garcini leaf and flower and its constituents have excellent antibacterial activity and are beneficial to human health. They have the potential to be used for medical purposes and to be utilized as antibacterial additives in making paper products. However, we need more investigation in vitro and in vivo. Keywords: Anvillea garcini, Streptococcus pyogenes, Staphylococcus saprophyticus, Enterobacter aerogenes, Bacillus anthracis doi:10.1016/j.clinbiochem.2011.08.873
Poster — [A-10-1154-2] Recombinant production of anti-HIV protein, Griffithsin, in E. coli Afshar Shandiz Mohammada, Vafaee Yavarb, Babalar Mesbahb, Alizadeh Houshanga a Agronomy Dep. of Agricultural Biotechnology, Agricultural College, Tehran University, Karaj, Iran b Dept. of Horticultural Science, Agricultural College, Tehran University, Karaj, Iran E-mail addresses: [email protected] (A.S. Mohammad), [email protected] (V. Yavar), [email protected] (B. Mesbah), [email protected] (A. Houshang) Over the past 27 years, nearly 25 million people have died from AIDS. Available strategy such as highly active antiretroviral therapy (HAART) can prevent HIV transmission from parents to children but fail to treat this disease. The red alga Griffithia sp. has a novel antiHIV protein which is called Griffithsin (GRFT). The potent anti-viral activity of GRFT against both laboratory and primary isolates of HIV at picomolar concentrations makes this protein an attractive candidate microbiocide to prevent sexual transmission of HIV. At the first step, we optimized codon usage and designed hexa-histidine tagged GRFT (His-GRFT) sequence for expression in Escherichia coli. Then synthesized GRFT sequence was inserted in expressional vector pET28a (+) and transformed to E. coli BL21 strain. Finally, the recombinant production of GRFT was confirmed by SDS-PAGE. The expression system described in this work can provide a basis for the mass production of GRFT to allow further studies of the protein and investigation of therapeutic and preventive strategies against HIV. Keywords: Griffithsin, Lectin, Anti-HIV, His tag, E. coli doi:10.1016/j.clinbiochem.2011.08.874
Poster — [A-10-1156-1] The effect of combination of aerobic training and oral administration of hydro extract of saffron (Crocus sativus) on serum lipid profile in streptozotosin induced diabetic rats Roya Irajia, Mohamad Ali Azarbayjanib, Mohamad Javad Rasaeec, Peeri Maghsooda, Asghar Nikpoora, Ayati Esfandyara a Ghods sq, Iranzamin St, Islamic Azad University, Central Tehran Branch Faculty of Aerobic Education, Tehran, Iran