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Phylogenetic Analysis of Lysozyme C from the Scorpion Mesobuthus eupeus Venom Gland
Ghafar Eskandari,Abbas Jolodar,Nabiallah Khoonmirzaei
Research Journal of Biological Sciences , 2012, DOI: 10.3923/rjbsci.2011.9.11
Abstract: Many studies have been carried out on peptides and genes encoding scorpion toxins from the venom of the scorpion Mesobuthus eupeus. The scorpion venom contains a diversity of bioactive peptides which could cause toxic effects and can be candidates for drug design and development. The anti-microbial lysozymes among them are of great value. Lysozymes are hydrolytic enzymes characterized by the ability to cleave the β-(1, 4)-glycosidic bond between N-acetylmuramic acid and N-acetyl-D-glucosamine in a peptidoglycan layer, the major bacterial cell wall polymer. The total RNA was extracted from venom glands of Mesobuthus eupeus species of Kuzestan. cDNA was synthesized with extracted total RNA as template and modified oligo (dT) as primer. In order to amplify cDNA encoding a Lys-C peptide, semi-nested RT-PCR was performed with the specific primers followed by sequencing of the amplified fragment. The full-length cDNA sequence contains a 438 nucleotide open reading frame encoding a peptide of 144 amino acids with molecular weight of 16.702 kDa. A putative 22-residue signal peptide was identified. Based on the phylogenetic tree of MesoLys-C and c-type lysozyme of East Mediterranean M. eupeus it is concluded that M. eupeus of Khuzestan and East Mediterranean M. eupeus belong to different subspecies.
Biochemical and Histopathological study of Mesobuthus eupeus scorpion venom in the experimental rabbits  [cached]
Zayerzadeh, E.,Zare Mirakabadi, A.,Koohi, M.K.
Archives of Razi Institute , 2011,
Abstract: In tropical and subtropical countries, envenomation by scorpions (so-called scorpionism) represents a serious public health problem. In the present study, the toxic effects of mice LD50 injections of Mesobuthus eupeus (Me) venom on the kidney and liver of anesthetized rabbits were investigated. Six rabbits were selected and ALT, AST, BUN and creatinine were measured at 0, 1 and 3 hours after envenomation and histopathological studies were carried out postmortem. All the animals showed signs and symptoms ofenvenomation within 30-40 minutes and died 3 to 3.5 hours after venom injection. Histopathological examinations revealed glumerolar congestion, dilated vessels of interstitium and focal interstitial congestion in the kidney and focal hemorrhage, central vein congestion, congested vessels in portal areas and dilatedsinusoids in the liver at 3 to 3.5 hrs following venom injection. In addition, biochemical analyses indicated significant rise in the levels of ALT and creatinine following Mesobuthus eupeus envenomation in animals at 3 hrs. However no significant changes were observed at 1 hr. In conclusion, scorpion (Mesobuthuseupeus) venom leads to damage in vital organs such as liver and kidney.
Cardiopulmonary complications induced by Iranian Mesobuthus eupeus scorpion venom in anesthetized rabbits
Zayerzadeh, E;Koohi, MK;Zare Mirakabadi, A;Purkabireh, M;Kassaaian, SE;Rabbani, SH;Sotoudeh Anvari, M;Boroumand, MA;Sadeghian, S;
Journal of Venomous Animals and Toxins including Tropical Diseases , 2010, DOI: 10.1590/S1678-91992010005000001
Abstract: scorpion envenomation is a life-threatening condition, especially in children and elderly individuals affected by respiratory and cardiovascular diseases. in this study, the toxic effects of median lethal dose (ld50) injections of mesobuthus eupeus (me) venom on the heart and lungs of anesthetized rabbits were investigated. six rabbits were selected and alterations in their electrocardiogram, heart rate, respiration and blood pressure before and after venom injection were recorded. cardiac troponin t (ctnt), creatinine kinase muscle-brain fraction (ck-mb) and lactate dehydrogenase (ldh) were measured at 0, 1 and 3 hours after envenomation and pathology studies were carried out postmortem. all the animals showed signs and symptoms of envenomation within 40 minutes and died 3 to 3.5 hours after venom injection. pathology studies revealed alveolar edema in 100% of the rabbits and myocardial infarction in 16%. the main histopathological changes were myocytolysis, coagulation necrosis, focal hemorrhage, thrombus formation both in myocardium and on endocardial surfaces as well as inflammatory infiltrates in the heart and hemorrhage, vascular thrombus and interstitial inflammation in the lungs. ecg monitoring of rabbits showed st elevation, st depression and inverted t and q waves. in addition, although ctnt levels increased in 16% of the animals and serum ldh was also augmented, none of these changes was statistically significant. the enzyme ck-mb also did not show any change after me venom injection. in conclusion, the results of this study showed that me venom killed animals in less than 3.5 hours through severe pulmonary damage and it appears that the deaths could not be attributed to cardiovascular lesions. therefore, me venom effects on the lungs are so important that they appear to be independent of heart damage.
Comparison of two purified toxic fractions from Mesobuthus eupeus scorpion venom
Shirmardi, SP;Shamsaei, M;Gandomkar, M;Saniei, E;Ghannadi, M;Zare, A;
Journal of Venomous Animals and Toxins including Tropical Diseases , 2010, DOI: 10.1590/S1678-91992010000400015
Abstract: iranian scorpions belong mainly to the buthidae and scorpionidae families, distributed into 16 genera and 25 species. in iran, similar to other parts of the world, there are a few known species of scorpions responsible for severe envenoming; amongst which mesobuthus eupeus is the most common. its venom contains several toxin fractions that may affect the ion channel. in the present study purification, labeling and biological evaluation of m. eupeus venom are described. for separation, soluble venom was loaded on a chromatography column packed with sephadex g-50 gel. subsequently, the fractions were collected according to uv absorption at a wavelength of 280 nm. toxic fraction (f3) was loaded on an anionic ion exchanger resin and then on a cationic resin. finally, toxic subfractions f3.1.6 and f3.1.9 were labeled with 99mtc and injected into normal mice to distinguish excretion pathway. the venom toxic fraction was successfully obtained in its purified form. radiolabeling of toxic fractions was performed at high specific activity with radiochemical purity of more than 97 and 95% respectively for f3.1.6 and f3.1.9. biodistribution studies in normal mice with two toxic fractions usually show rapid clearance of the compounds from blood and tissue except for kidneys. since tissue distribution studies are very important for clinical purpose, the present findings suggest that 99mtc labeling of venom is a useful tool for in vivo studies and comprises an excellent approach to monitoring the process of biodistribution and kinetics of toxins.
Preparation and Biodistribution Study of a 99mTc-Labeled Toxic Fraction of Iranian Mesobuthus Eupeus Scorpion Venom
Seyed Pezhman Shirmardi,Mostafa Gandomkar,Mojtaba Shamsaei,Abbas Zare Mirakabadi
Iranian Journal of Nuclear Medicine , 2010,
Abstract: Introduction: Iranian scorpion species are classified in Buthidae and Scorpionidae with 16 genera and 25 species. In Iran, similar to other parts of the world, there are a few known species of scorpions responsible for severe envenoming. Mesobuthus eupeus is the most common species in Iran. Its venom contains several toxin fractions which can affect the ion channel. In this study purification, labeling and biological evaluation of Mesobuthus eupeus scorpion venom are described. Methods: To separate different venom fractions, soluble venom was loaded on a chromatography column packed with sephadex G50 gel then the fractions were collected according to UV absorption at 280 nm wavelength. Toxic fraction (F3) was loaded on anionic ion exchanger resin (DEAE) and then on a cationic resins (CM). Finally toxic fraction F319 was labeled with 99mTc and radiochemical analysis was determined by paper chromatography. The biodistribution was studied after injection into normal mice. Results: Toxic fraction of venom was successfully obtained in purified form. Radiolabeling of venom was performed at high specific activity with radiochemical purity more than 95% which was stable for more than 4 h. Biodistribution studies in normal mice showed rapid clearance of compound from blood (2.64% ID at 4 h) and tissues except the kidneys (27% ID at 4 h). Conclusion: As tissue distribution studies are very important for clinical use, results of this study suggest that 99mTc labeling of venom can be a useful tool for in vivo studies and is an excellent approach to follow the process of biodistribution and kinetics of toxins.
Sequence Analysis of Txk from the Scorpion Mesobuthus eupeus Venom Glands Using Semi-Nested RT-PCR
Ghafar Eskandari,Abbas Jolodar,Ahmad Taghavi Moghadam
Research Journal of Biological Sciences , 2012, DOI: 10.3923/rjbsci.2011.12.14
Abstract: Natural toxins are useful probes for evaluating the involvement of K+ channels in cell activity and for investigating K+ channel structure and localization. In recent years, peptide toxins that block various K+ channels with high affinity have been purified from diverse animal venoms. One polypeptide beta neurotoxin named Txk was isolated from the venom of scorpion Mesobuthus eupeus of Khuzestan. This toxin consists of 91 amino acid residues which modulate voltage-gated sodium channels gating. In this study, cDNA of Txk β-toxin was amplified and sequence of beta neurotoxin compared with M. martensii occitanus Israelis and Tityus costatus however, the comparison suggests that the length of the peptide is close to the long-chain potassium ion channel blocker peptide family.
Characterization of cDNA sequence encoding for a novel sodium channel -toxin from the Iranian scorpion Mesobuthus eupeus venom glands  [cached]
Eskandari, Gh.,,Jolodar, A.,Ktsoyan, Z.,Seyfiabad Shapouri, M.R.
Archives of Razi Institute , 2012,
Abstract: The venoms of Buthidae scorpions are known to contain basic, single-chain protein -toxins consisting of 60-70 amino acid residues that are tightly cross-linked by four disulfide bridges. Total RNA was extracted from the venom glands of scorpion Mesobuthus eupeus collected from the Khuzestan province of Iran and then cDNA was synthesized with the modified oligo (dT) primer and extracted total RNA as template. By applying the Semi-nested RT-PCR using homologous primers, the fragments of 273 bp was amplified and sequenced. The coding region encodes a polypeptide of 85 amino acid residues with a calculated molecular weight of 9.337 kDa and theoretical isoelectric point of 6.52. It was designated as MeNaTx -4. This peptide with homology to the "Scorpion long chain toxin-like domain" is belongs to the Toxin-3 superfamily. Multiple alignment of the putative amino acid sequence of MeNaTx -4 exhibited 95% sequence identity with lesser Asian scorpion M. eupeus venom sodium channel -toxin-3 but interestingly displayed only 63% identity with other -toxins from this species. The sequence was also identical to the counterpart sequences from M. martensii (92%) and Buthus occitanus israelis (80%). This high sequence similarity together with completely conserved in the positions of all 8 Cys indicated that MeNaTx -4 is a new member of -toxin from the Iranian M. eupeus which may originate from a common ancestor.
Preparation and investigation of a toxic fraction of mesobuthus eupeus labeled with 99mTc
Seyed Pezhman Shirmardi,Mostafa Gandomkar,Mojtaba Shamsaei,Abbas Zare Mirakabadi
Iranian Journal of Nuclear Medicine , 2010,
Abstract: Introduction: Iranian scorpion species are classified in Buthidae and Scorpionidae with 16 genera and 25 species. In Iran, similar to other parts of the world, there are a few known species of scorpions responsible for severe envenoming. Mesobuthus eupeus is the most common species in Iran. Its venom contains several toxin fractions which can affect the ion channel. In this study purification, labeling and biological evaluation of Mesobuthus eupeus scorpion venom are described. Methods: To separate different venom fractions, soluble venom was loaded on a chromatography column packed with sephadex G50 gel then the fractions were collected according to UV absorption at 280 nm wavelength. Toxic fraction (F3) was loaded on anionic ion exchanger resin (DEAE) and then on a cationic resins (CM). Finally toxic fraction F319 was labeled with 99mTc and radiochemical analysis was determined by paper chromatography. The biodistribution was studied after injection into normal mice. Results: Toxic fraction of venom was successfully obtained in purified form. Radiolabeling of venom was performed at high specific activity with radiochemical purity more than 95% which was stable for more than 4 h. Biodistribution studies in normal mice showed rapid clearance of compound from blood (2.64% ID at 4 h) and tissue except the kidneys (27% ID at 4 h). Conclusion: As tissue distribution studies are very important for clinical use, results suggest that 99mTc labeling of venom can be a useful tool for in vivo studies and is an excellent approach to follow the process of biodistribution and kinetics of toxins
In vivo and in vitro effects of scorpion venoms in Turkey: a mini-review
Adiguzel, S;
Journal of Venomous Animals and Toxins including Tropical Diseases , 2010, DOI: 10.1590/S1678-91992010000200004
Abstract: scorpion stings constitute a common type of accident in turkey due to the geographic location, climate and socioeconomic structure of this country. moreover, envenomation cases are considered a public health problem throughout turkey. based on data in the literature, the main clinical features of scorpion envenomations are defined as systemic manifestations including sweating, hypertension and vomiting, and among prominent causes of death are cardiorespiratory manifestations, toxic myocarditis and pulmonary edema. the most important health-threatening scorpions found in turkey are: androctonus crassicauda, leiurus quinquestriatus, mesobuthus gibbosus and mesobuthus eupeus, all of which belong to the buthidae family. this study indicates that there is no data related to venom toxicity and in vivo effects of other scorpions found in turkey - such as mesobuthus caucasicus, mesobuthus nigrocinctus and hottentotta saulcyi - and that further research should be performed to determine their health effects.
The action of red scorpion (Mesobuthus tamulus coconsis, pocock) venom and its isolated protein fractions on blood sodium levels
Badhe, R. V.;Thomas, A. B.;Deshpande, A. D.;Salvi, N.;Waghmare, A.;
Journal of Venomous Animals and Toxins including Tropical Diseases , 2007, DOI: 10.1590/S1678-91992007000100006
Abstract: red scorpion (mesobuthus tamulus or buthus tamulus) venom samples were collected at different regions of india: western (chiplun and ahmednagar from maharashtra state) and southern (ratnagiri and chennai from tamil nadu state). the action of whole venoms on the blood sodium levels of mice was assessed using flame photometry. seven peptides were common to all venom samples. they were separated using the native polyacrylamide gel electrophoresis (page) technique and their activities were also studied using flame photometry. there was a decrease in the concentration of sodium ions in the serum, which suggested the blockage of such ions by scorpion venom toxins. among the 10 protein bands isolated, the band at 79.6 kda presented maximum activity in decreasing serum sodium ions concentration. whole venom from chiplun region also showed maximum activity. the western blotting technique demonstrated that the anti-scorpion venom sera produced by haffkine biopharmaceuticals corporation ltd., india, neutralized all four venom samples.
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