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Proteases from Cerastes cerastes and Cerastes vipera venoms digested casein, gelatin and hemoglobin. Gelatin digestion was detected by reacting the trichloroacetic acid soluble peptides with trinitrobenzene sulfonate. This method is more sensitive than ultraviolet absorption, and is applicable to protein substrates irrespective of their aromatic amino acid content. The proteases showed optimal pH values of 7·8 for casein and 7·0 for hemoglobin digestion. They were activated by Ca2+, Ba2+ and Mg2+, and were completely inhibited by 1 mM ethylenediamine tetra-acetate. This inhibition could be reversed maximally (73%) by 3 mM Ca2+. Electrophoresis at pH 6·0 resolved the proteases into a major band migrating very slowly towards the cathode (3 mm/hr) and a minor more rapidly migrating band (14 mm/hr). Sephadex G—100 gel filtration resolved the proteolytic activity into 4 peaks in each venom. Their estimated mol. wts were 120,000, 49,000, 23,000 and 16,000 in C. cerastes and 130,000, 56,000, 25,000 and 14,000 in C. vipera. The first peak in both venoms showed the highest protease activity and was also the most lethal, causing marked hemorrhages in mice. 相似文献
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R. S. Labib, H. Y. Halim and N. W. Farag. Fractionation of Cerastes cerastes and Cerastes vipera snake venoms by gel filtration and identification of some enzymatic and biological activities. Toxicon17, 337–345, 1979.—Cerastes cerastes and Cerastes vipera venoms were fractionated by gel filtration on Sephadex G-100. The gel filtration patterns were similar, each showing eight fractions. The molecular weights and Stokes radii (in Å) of l-amino acid oxidase, phosphodiesterase, hyaluronidase (main peak) and phospholipase A were 150,000, 51·7 Å, 105,000, 44·2 Å, 75,000, 37·5 Å, and 22,000, 22·7 Å for C. cerastes, and 150,000 51·7 Å, 95,000, 41·8 Å, 72,000, 36·3 Å and 17,000, 20·2 Å for C. vipera, respectively. Most of the fractions were lethal, but the first fraction (pool A, molecular weight above 75,000) in both species, was the most lethal (MLD = 3 mg per kg) and accounted for two thirds or more of the recovered lethality. Hemorrhage in the abdominal wall and mesenteries was characteristic for this fraction. Another fraction (pool C, molecular weight 40,000–60,000, C. cerastes venom) was characterized by rapid death, and its MLD was 7 mg per kg. At a dose of 15–20 mg per kg, this fraction killed mice in 1–5 min when injected i.p. while the first fraction (pool A) killed in 1–3 hr. 相似文献
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A coagulant component has been purified from Cerastes cerastes venom, using gel filtration on a Sephadex G 100 (fine) column followed by chromatography on a Bio-Rex 70 column. This compound had a proteolytic effect and could coagulate human plasma deficient in factor VIII or (VIII + IX) with the formation of a firm clot. It could also clot plasma deficient in factor X, but the clot formed was soft and not complete. The compound had no effect on platelet aggregation and was nontoxic. This compound is believed to be primarily a factor X activator, as it could replace factor VIII in hemophilic plasmas. 相似文献
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Zaineb Abdelkafi-Koubaa Ines ELBini-Dhouib Soumaya Souid Jed Jebali Raoudha Doghri Najet Srairi-Abid Khadija Essafi-Benkhadir Olivier Micheau Naziha Marrakchi 《Toxins》2021,13(12)
Snake venom proteins, which are responsible for deadly snakebite envenomation, induce severe injuries including neurotoxicity, myotoxicity, cardiotoxicity, hemorrhage, and the disruption of blood homeostasis. Yet, many snake-venom proteins have been developed as potential drugs for treating human diseases due to their pharmacological effects. In this study, we evaluated the use of, an L-amino acid oxidase isolated from Cerastes cerastes snake venom CC-LAAO, as a potential anti-glioblastoma drug, by investigating its in vivo and in vitro pharmacological effects. Our results showed that acute exposure to CC-LAAO at 1 and 2.5 µg/mL does not induce significant toxicity on vital organs, as indicated by the murine blood parameters including aspartate transaminase (AST), alanine transaminase (ALT), lactate dehydrogenase (LDH) activities, and creatinine levels. The histopathological examination demonstrated that only at high concentrations did CC-LAAO induce inflammation and necrosis in several organs of the test subjects. Interestingly, when tested on human glioblastoma U87 cells, CC-LAAO induced a dose-dependent apoptotic effect through the H2O2 generated during the enzymatic reaction. Taken altogether, our data indicated that low concentration of CC-LAAO may be safe and may have potential in the development of anti-glioblastoma agents. 相似文献
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An 18 year old woman was bitten on the second finger of her right hand by a Cerastes cerastes. A literature search revealed very few clinical cases, and most of those were only laboratory and in 'vitro studies'. Local signs included a hemorrhagic wound at the site of the bite with marked swelling of the entire hand. Twelve hours later, the patient developed coagulopathy, which lasted 3 days and required repeated administration of blood products. Treatment was essentially supportive and the patient was discharged from the hospital after 5 days in good condition. 相似文献
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A single phospholipase A2 has been found in Cerastes cerastes venom, purified to homogeneity by a combination of chromatographic steps involving gel filtration on Sephadex G-50 and ion-exchange chromatography on DEAE-Sephadex A-50. Its mol. wt, its amino acid composition and its partial amino acid sequence have been determined. High homologies between its sequence and those of other Viperid phospholipides A2 have been noticed. The phospholipase was non-lethal to mice up to a dose as high as 25 mg/kg by i.p. and i.v. injection. This non-toxic enzyme exhibited an acidic isoelectric point and hydrolyzed monolayers of different short chain phospholipids. Some kinetic parameters have been studied potentiometric titration (with or without Triton X-100) and the rate of catalysis seemed not to be affected by changes in the physical state of the substrate. 相似文献
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The aim of this study was to obtain anti-snake antiserum by optimizing the conditions of extraction and purification and test its ability to neutralize local myonecrosis. Extraction and purification was achieved through adjustment of the pH, pepsin concentration, time of digestion, and caprylic acid concentration. Our results indicate that the best conditions to obtain anti-snake antiserum from ammonium sulfate fractionated plasma are pH 3.3, 3.5 g/l pepsin, digestion for 90 min at 37 degrees C, and 0.5% caprylic acid. Antiserum purified using this method has greater neutralizing ability of myonecrosis than ammonium sulfate (ammSO4) fractionated product. 相似文献
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Purification and characterization of phosphodiesterase (exonuclease) from Cerastes cerastes (Egyptian sand viper) venom 总被引:1,自引:0,他引:1
A venom exonuclease 'phosphodiesterase' (E.C. 3.1.4.1) has been purified from Cerastes cerastes venom by a combination of gel filtration on Sephadex G-100 superfine and ion exchange chromatography on DEAE-Sepharose. The enzyme showed a single band on PAGE and SDS-PAGE and had a molecular weight of 110,000. The final preparation was purified 28 fold. It had no carbohydrate and it did not have protease or 5'-nucleotidase activities. Optimum temperature for enzyme activity was 56 degrees C. The enzyme was rapidly inactivated when pre-incubated above 40 degrees C. Energy of activation (Ea) was calculated to be 0.913. The optimum pH was 9.0. Cysteine, glutathione, dithiothreitol, 2-mercaptoethanol, ADP and AMP inhibited the enzyme. Cysteine caused a non-competitive inhibition, while ADP showed a competitive inhibition. EDTA at a concentration of 0.5 mM caused complete inhibition of the enzyme, which could be reversed by the addition of Ca2+ or Mn2+. 相似文献
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A A Mustafa 《Immunopharmacology》1990,20(2):115-123
Cerastes cerastes venom added in vitro to human whole blood caused a marked inhibitory effect on the luminol-dependent chemiluminescence induced by phorbol myristate acetate (PMA) or opsonized zymosan on phagocyte cells. The inhibitory effect produced by the venom was both dose- and time-dependent when PMA was used as the stimulant of the oxidative burst. Similarly, the venom produced a significant inhibitory effect when added to isolated polymorphonuclear leukocytes (PMNs). Incubation of the isolated PMNs with the highest concentration of the venom used (1000 micrograms/ml), however, did not result in any significant disruption of the membranes of these cells. The effect of the venom on the isolated PMNs was also reversible following washing of the venom-treated cells with phosphate-buffered saline. The scavenger of reactive oxygen species such as superoxide dismutase, catalase and dimethyl sulphoxide potentiated the effect of the venom on the luminol-dependent chemiluminescence of human phagocyte cells. Sodium azide, the myeloperoxidase inhibitor, and sodium benzoate produced a similar potentiating effect. The results suggest that some of the toxicity of the venom may be mediated by an action on the phagocytic immune system. 相似文献
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Envenomation by snake venoms would be expected to result in proteolysis of plasma proteins as well as of cellular constituents. Incubation of human serum with crude venom from Cerastes cerastes showed that the plasma lipoproteins were a target of this venom. Fractionation of the crude venom by gel filtration revealed that high density lipoprotein (HDL) was susceptible almost exclusively to the highest mol. wt fraction of venom and that proteolysis was due to a metalloprotease. Although HDL was degraded only by this metalloprotease, the low density lipoprotein (LDL) was proteolyzed by both metalloproteases and serine proteases present in several fractions of the venom. Despite extensive degradation, LDL remained intact, as judged by gradient gel electrophoresis. The selectivity of venom fractions may prove useful in the study of lipoprotein structure. 相似文献
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The effects of the venom of the sand viper (Cerastes cerastes) on haematological and cardiovascular parameters and on isolated ileum, trachea, pulmonary artery and atrium from the guinea-pig were studied. In concentrations from 0.2 micrograms/ml to 0.6 mg/ml, snake venom caused concentration-dependent relaxation of the longitudinal ileal segments and the epinephrine-precontracted pulmonary artery rings, increased the tone of the tracheal rings and increased the rate of the spontaneously beating atrium but depressed the amplitude of its contraction. Low doses (0.04-0.4 mg) of the venom caused transient small depression of both heart rate and contractility of the isolated perfused heart. A higher dose (1.0 mg) significantly inhibited both parameters and delayed their recovery. An i.v. injection of 0.12 mg/kg of the snake venom to anaesthetized guinea-pigs had no effect on red blood cell count, haemoglobin concentration or the haematocrit value but significantly reduced the circulating white blood cells, the plasma clotting time and erythrocyte deformability. Also, i.v. injections of 0.02-0.2 mg/kg of the venom caused initial depression of the systolic and the diastolic blood pressure followed by complete or partial recovery and subsequent hypotension. These observations indicate that C. cerastes venom has multiple sites of action which may help in better understanding the pathology of bites by this snake. 相似文献
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Cerastes cerastes venom in low concentrations (0.8-2.5 micrograms/ml) inhibited the response of the rat vas deferens to field stimulation, while higher concentrations (4-8 micrograms/ml) causes a contracture that was reversed by washing and transiently blocked by phentolamine. The effect of the venom and guanethidine (10(-7) M) was reversed by tyramine (4.6 x 10(-8) M) but unaffected by hexamethonium (1.4 x 10(-7) M). When the response to field stimulation was inhibited by the venom the response to norepinephrine was either potentiated or unchanged. 相似文献
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An enzyme displaying proteolytic activity toward the natural substrate casein as well as clotting activity on fibrinogen was purified to homogeneity from Cerastes cerastes (horned viper) venom and characterized. The enzyme is constituted of two identical subunits of mol. wt 48,500 as determined by SDS-polyacrylamide gel electrophoresis, and has an isoelectric point of 3.75. N-terminal sequencing up to the 33rd residue evidenced a high homology with other snake venom proteinases. The proteinase is of serine-type as indicated by high sensitivity to DFP and shows both arginine-ester hydrolase and amidase activities on synthetic substrates. Both specific activities were 30-fold higher than the respective activities found in the crude venom. The Km value determined for arginine-containing substrate BAEE was 3.0 x 10(-4) M and the Km for chromogenic substrate CBS 34-47 0.65 x 10(-4) M. The Vm/Km ratio, however, was two-fold higher for BAEE than for CBS 34-47; the arginine-esterase activity of this enzyme is thus slightly higher than its amidase activity. 相似文献
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Abhinandan Chowdhury Matthew R. Lewin Rebecca Carter Raul Soria Matt Aldridge Bryan G. Fry 《Toxins》2022,14(12)
The African viperid snake genera Atheris, Cerastes, and Proatheris are closely related, similar in size, but occupy extremely divergent ecological niches (arboreal in tropical rainforests, fossorial in deserts, and swamp-dwelling, respectively). Their venoms have not previously been subjected to comparative analyses for their action upon the coagulation of blood, most notably with significant data deficiencies from Atheris and Proatheris. In contrast, the closely related genus Echis is well-documented as capable of producing potent procoagulant effects. In light of this, we set out to compare the coagulotoxic actions of Atheris ceratophora, A. chlorechis, A. desaixi, A. nitschei, A. squamigera, C. cerastes, C. cerastes gasperettii, C. vipera, and Proatheris superciliaris and explore potential pharmacological interventions to reestablish normal blood coagulation. All venoms displayed extremely potent procoagulant effects, over twice as fast as the most potent Echis reported to date. Although Cerastes is used in the immunising mixture of two different regionally available antivenoms (Inoserp-MENA with C. cerastes, C. cerastes gasperettii, C. vipera and Saudi Arabian polyvalent with C. cerastes), none of the other species in this study are included in the immunising mixture of any antivenom. Notably, all the Cerastes species were only neutralised by the Inoserp-MENA antivenom. C. cerastes venom was not neutralised well by the Saudi Arabian antivenom, with the low levels of recognition for any of the Cerastes venoms suggesting a strong regional variation in the venom of this species, as the C. cerastes venom tested was of African (Tunisian) origin versus Saudi locality used in that antivenom’s production. The other antivenoms (Micropharm EchiTAbG, ICP EchiTAb-Plus-ICP, Inosan Inoserp Pan-Africa, Premium Serums PANAF Sub-Sahara Africa, South African Vaccine Producers Echis, South African Vaccine Producers Polyvalent) all displayed trivial-to-no ability to neutralise the procoagulant toxicity of any of the Atheris, Cerastes, or Proatheris venoms. Comparative testing of the enzyme inhibitors DMPS, marimastat, and prinomastat, revealed a very potent neutralising capacity of marimastat, with prinomastat showing lower but still significant potency at the same molar concentration, while a 5× molar concentration of DMPS had no apparent effect on procoagulant venom effects normalized by the other inhibitors. These results and methods contribute to the body of knowledge of potential clinical effects and data necessary for evidence-based advancement of clinical management strategies. 相似文献
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子宫中隔切除术后预防粘连方法探讨 总被引:1,自引:0,他引:1
目的 探讨子宫中隔并不孕患者宫腔镜术后不同处理方法预防宫腔粘连的效果.方法 55例子宫中隔并不孕患者行腹腔镜监护下宫腔镜子宫中隔切除术(TCRS),术后针对不同患者采用不同术后处理措施,包括宫腔放置与不放IUD,是否进行人工周期治疗,部分患者术后使用GnRH-a类药物治疗术后第1、3个月行宫腔镜检查随访,宫内放置IUD的患者;于术后第3个月取环.结果 54例患者术后进行宫腔镜检查随访,其中40例分别于术后第1、3个月完成了术后2次宫腔镜检查,另14例只完成一次检查.宫腔术后放环与否对术后宫腔形态影响无差异(P>0.05),术后使用人工周期治疗患者较未使用者子宫内膜厚,此两者术后第3个月宫腔镜检查发现宫底创面均已有内膜覆盖.使用GnRH-a类药物患者术后官腔形态满意.结论 TCRS术后宫腔放置IUD无助于预防术后粘连的发生;术后人工周期治疗应更个体化并有针对性的使用GnRH-a类药物治疗.术后及时进行宫腔镜检查随访可防止术后宫底新粘连的形成. 相似文献