Effect of crude and carboxyl-functionalized carbon nanotubes on protein–protein interaction

Abstracts

Khozestan (south western Iran) is one of the richest scorpion distributed proviences and these species from Buthidae, Scorpionidae and Heimiscorpionidae families. Most of scorpions recognized are belonged to Buthidae family. Apistobuthus susanae belonging to Buthidae family is yellow in colour, hunted from sandy substrate places and strands. Its mature length is more than 10 cm, its particular character is flared disc shaped second metasomal segment, a feature unique among all known scorpions. They were hunted in the field at night by ultra violet light. The estimated parameters in venom are amount of protein (Lowry method), LD50, and gel electrophoresis, determination of phospholipase A2 activity (Oung Shain method), and determination of hemolysis percent (Ceul Smith method). The PH of the venom was near neutral and 80% of venom was its proteins, LD50 of venom 36 μg, had 12 protenic bands in electrophoresis. This scorpion uses its neurotoxin during hunt to immobilize the insects and also uses its venom as a defence mechanism against predators.

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changes such as necrosis with edema, inflammatory cell infiltration in the parenchymal and vascular space around the portal areas were observed. Amount of mercury (μg Hg/g tissue) in the rat liver (1 g) by AAS in group II (47.78 ± 7.80) showed accumulation about 13.15% than group I, and this case in group IV was 6.92% (33.10 ± 6.01) than group II, adjusted (P < 0.05). Administration of the EJFE to rats as IP afforded a significant protection against HgCl2 induced hepatotoxicity in all these diagnostic parameters. Conclusion: According to the results, use of this plant flowers extract increases antioxidant levels and through scavenging free radicals (ROS) performance, toxic metals' chelation have protective activity on hepatotoxicity and increased GSH levels in the body can be effective in reducing oxidative stress induced by toxins, drugs, and toxic metals. Keywords: Toxic metals, Mercuric chloride, Liver toxicity, Eriobotrya japonica, Antioxidant

Keywords: Scorpion, Venom, Buthidae, Apistobuthus doi:10.1016/j.clinbiochem.2011.08.893 doi:10.1016/j.clinbiochem.2011.08.892

Poster – [A-10-155-2] Protective effect of the Eriobotrya japonica flower extract (EJFE) on liver toxicity induced by mercuric chloride in male wistar rats Esmaeili Amirhosseina, Hajizadeh Moghaddam Akbarb, Chaichi Mohammad Javada, Ebrahim-Zadeh Mohammad Alic, Parsian Hadid a Department of Chemistry, Mazandaran University, Babolsar, Iran b School of Biology, Mazandaran University, Iran c School of Pharmacy, Medical University of Mazandaran, Sari, Iran d Department of Biochemistry, Medical University of Babol, Iran E-mail addresses: [email protected] (E. Amirhossein), [email protected] (H.M. Akbar), [email protected] (C.M. Javad), [email protected] (E.-Z.M. Ali) Introduction: Exposure to toxic and heavy metals such as Pb, Cd, and Hg can lead to chronic liver diseases including liver cirrhosis. Today, the use of herbal extracts with chelating activities, antioxidant, restoration of GSH level and no side effects can be effective in treating and control damage caused by toxic metals. In this study, the effects of hydroalcoholic extract of EJFE were investigated in order to protect the liver from damage caused by mercury toxic metal as in vivo. Materials and methods: The hydroalcoholic extract of EJFE used were freshly prepared. Adult male rats used ranged 180 ± 20 g with free access to water and standard food and were divided into six groups: group I (normal control), group II (mercury stress with 5 mg Hg/kg b.w. for about one month was added in drinking water), group III (normal control under the administration of extract 200 mg EJFE/kg b. w. rate three times a week for about one month as IP) and group IV (stress group with mercury were like group III under the administration of EJFE). Amount of MDA, Total Antioxidant Capacity (TAC), GSH level and the level of ALT, AST and ALP were determined in rat liver homogenized and plasma. Pathological effects of mercury on rat liver and protective effect of extracts of the plant were analyzed after sampling and staining with Hematoxylin–Eosin. Also, mercury levels in different groups of liver tissue were measured by AAS. Statistical analysis: All data were expressed as Mean ± SD and ANOVA was used to compare values between groups with P < 0.05 level was considered significant. Result: In this study, oral administration of 5 mg/kg b.w. HgCl2 induced liver damage evident as elevation in plasma ALT, AST, ALP (IU/L) and TAC (μmol/L) with FRAP method in plasma decreased. In rat liver homogenate (1 g) levels MDA (nmol MDA/g tissue) was used, TBARS method increased but levels of GSH (nmol GSH/mg protein) according to Ellman method and TAC decreased. Histopathological

Poster — [A-10-182-1] Biochemistry study of nitrate effect in drinking water on human hemoglobin Rezan Amjadi, Ghourchian Hedayatollah, Ali Akbar Moosavi-Movahedi Institute of Biochemistry and Biophysics, Tehran University, Tehran, Iran E-mail addresses: [email protected] (R. Amjadi), [email protected] (G. Hedayatollah), [email protected] (A.A. Moosavi-Movahedi) Introduction: Nitrate in drinking water is a health hazard because of its conversion to nitrite in saliva and gastrointestinal tract. High enough concentrations of nitrate in drinking water can result in a blood disorder called methemoglobinemia (metHb). The interaction of nitrite with human oxyhemoglobin (oxyHb) was investigated by optical spectroscopy and circular dichorism (CD). The nitrite ion oxidizes the Fe2+ of oxyhemoglobin to Fe3+ of methemoglobin. Circular dichorism studies show that interaction of nitrite with oxyHb decreases the alpha helix percentage and increases the beta sheet percentage at various concentration of nitrite indicating a conformational change of the protein. Conclusion: Optical spectra studies indicate the blue shift soret bond at 35 ppm of nitrite concentration probably destruction of heme center of this protein. Keywords: Methemoglobin, Nitrite, Drinking water doi:10.1016/j.clinbiochem.2011.08.894

Poster — [A-10-210-1] Effect of crude and carboxyl-functionalized carbon nanotubes on protein–protein interaction Mohammad-Bagher Ebrahim-Habibi, Majid Zeinali, Majid Soheili Biotechnology Research Center, Research Institute of Petroleum Industry (RIPI), West Blvd. of Azadi Sport Complex, Tehran, Iran E-mail addresses: [email protected] (M.-B. Ebrahim-Habibi), [email protected] (M. Zeinali), [email protected] (M. Soheili) Introduction: Recently, considerable effort has been generated to characterize intermediate structures accumulated during protein folding and unfolding. Growth of toxic amyloid structures, is supposed to be associated with various late-onset neurodegenerative disorders, initiated from intermediate structures interaction. The potential of nanoparticles to promote protein fibrillation is demon-

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Abstracts

strated. In the present study, acidic fibrillation of hen–egg weight lysozyme (HEWL) was analyzed in the presence of crude and carboxyl-functionalized nanotubes. Methods: Multi-walled CNTs were acid oxidized to form carboxylic acid group on the surface of the CNTs. IR-spectroscopy was used to verify the CNT functionalization. A sample solution of 2 mg/ml HEWL in acidic water with salts was prepared. Nanotubes at two final concentrations were added to HEWL solution and were incubated at 55 °C at different times. Congo red binding assay and Far-UV CD spectroscopy were used to characterize amyloid fibrillation. Results and discussion: Results from Congo red absorption spectroscopy demonstrated the formation of fibrillar amyloid structures at 72 h after the initiation of HEWL incubation. In addition, CD spectra of HEWL exhibited a characteristic pattern of β-sheet conformation. In exploring if CNT exerted a stimulatory action on the formation of HEWL fibrils, we monitored the changes of Congo red binding and CD spectra in amyloidogenic conditions. Our observations indicated that crude CNT could outstandingly enhance amyloid fibril formation while CNT in carboxylated form had no significant effect on fibrillation process. Conclusion: It is concluded that introduction of polar groups on the hydrophobic surfaces may play an important inhibitory role on surface-assisted nucleation. Keywords: Carbon nanotube, Protein folding, Fibrillation, Carboxylation doi:10.1016/j.clinbiochem.2011.08.895

Poster — [A-10-225-1] Cytotoxic potential of five Iranian medicinal plants by Artemia urmiana Mehdi Ghahremanifar, Ali Mirzaei, Maasoomeh Kazemi Yasouj Medical Sciences University, Medical College, Yasouj, Iran E-mail addresses: [email protected] (M. Ghahremanifar), [email protected] (A. Mirzaei), [email protected] (M. Kazemi) Introduction: Many plant products have strong anticancer activity against cancer cells (human and rodents). Phytochemical compounds in plants causing to strong antioxidant activity that may prevent from some cancers or some other diseases through inhibition of free radicals. Thus, antioxidant-rich plants can cause health and prevent from growing of cancer. One of the most reliable tests is the brine shrimp lethality test (BLST) that is simple procedure for screening of plant toxicity and bioactive compounds having antitumor activity. Material and methods: In the present study, aqueous extracts of Mentha logifolia, Achillea mellefolium, Mentha piperita, Matricaria recutita and Teucrium polium medicinal plants were screened for their cytotoxicity using brine shrimp lethality test. Five concentrations (25, 50, 75, 100, and 150 ppm) of each extract were applied in the bioassay and the LC50 determined. Results: The least LC50 value of 32 was obtained for hexane extract and the highest value of 13.87 was obtained for aqueous extract using the Finneys probit analysis which indicates high lethality of the extracts and presence of potent cytotoxic compounds in the plant extracts. Out of the 5 plants tested, M. recutita exhibited potent brine shrimp lethality with LC50 32 μg. M. logifolia, A. mellefolium, T. polium and M. piperita have also showed moderate cytotoxicity with LC50 111, 160, 172 and 247 μg respectively. Conclusion: The present study supports that brine shrimp bioassay is simple reliable and convenient method for evalution of bioactivity of medicinal plants and lends support for their use in traditional medicine.

Keywords: Artemia urmiana, Brine shrimp lethality test, Medicinal plants, Cytotoxicity doi:10.1016/j.clinbiochem.2011.08.896

Poster — [A-10-265-1] Beneficial effects of allicin on lead induced tissue enzyme alterations in common carp (Cyprinus carpio) Hasan Baghshani, Davar Shahsavani Ferdowsi University of Mashhad, Mashhad, Iran E-mail addresses: [email protected] (H. Baghshani), [email protected] (D. Shahsavani) Introduction: In toxicological studies, the alterations in the enzymatic activities directly reflect the metabolic disturbances and cell damage in specific organs. The aim of the current study was to determine the effects of lead exposure on tissue enzyme activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) in selected tissues of common carp and also to evaluate the beneficial effects of allicin, the main active constituent of garlic, on modulating enzyme alterations. Methods: Common carp (Cyprinus carpio; total, n = 60) were divided randomly into three groups of 20 each. Group I fish were reared in normal freshwater and served as the control. Group II fish were exposed to lead acetate (7.0 mg L−1) for 10 days. Group III fish were exposed to lead acetate (7.0 mg L−1) for 10 days and also received dietary allicin (10 mg/kg body weight, daily) during lead acetate treatment. At the end of each exposure, enzyme activities in prepared tissue homogenates were determined using colorimetric methods. Results: Lead exposure caused a significant elevation of AST, ALT and LDH in the liver, as compared to control group (P < 0.05). The AST, ALT, ALP and LDH levels in kidney homogenates were considerably increased following lead exposure, although these increases were only significant for ALT and ALP values. Significant increases of brain ALT and gill ALP activities were also observed following lead exposure. However, alterations in the activities of other measured enzymes in both brain and gill of experimental groups were not significant. Conclusion: Allicin supplementation was effective in attenuating the increase in Pb-induced tissue enzyme activities in the most organs; therefore it might have some protective and therapeutic effects on lead poisoning. However, much more extensive biochemical and toxicological research are needed for better understanding of the molecular basis of lead induced tissue enzyme alterations and ameliorative properties of allicin. Keywords: Lead poisoning, Allicin, Tissue enzyme activity, Common carp doi:10.1016/j.clinbiochem.2011.08.897

Poster — [A-10-274-1] Purification and immunogenic characterization of anti-LPS rabbit antibodies produced against Salmonella Typhimurium extracted lipopolysaccharide Ebrahimi Zadeh Waleada, Seyedlatif Mousavi Gargarib, Masoumeh Rajabi Bazlb, Mohammad Mohammadib a Shahed University, Qom, Iran b Tehran-qum Express Way, Shahed University, Iran E-mail addresses: [email protected] (E.Z. Walead), [email protected] (S.M. Gargari),