Understanding the complexity of Tityus serrulatus venom: A focus on high molecular weight components

Tityus serrulatus scorpion is responsible for a significant number of envenomings in Brazil, ranging from mild to severe, and in some cases, leading to fatalities. While supportive care is the primary treatment modality, moderate and severe cases require antivenom administration despite potential limitations and adverse effects. The remarkable proliferation of T. serrulatus scorpions, attributed to their biology and asexual reproduction, contributes to a high incidence of envenomation. T. serrulatus scorpion venom predominantly consists of short proteins acting as neurotoxins (α and ß), that primarily target ion channels. Nevertheless, high molecular weight compounds, including metalloproteases, serine proteases, phospholipases, and hyaluronidases, are also present in the venom. These compounds play a crucial role in envenomation, influencing the severity of symptoms and the spread of venom. This review endeavors to comprehensively understand the T. serrulatus scorpion venom by elucidating the primary high molecular weight compounds and exploring their potential contributions to envenomation. Understanding these compounds' mechanisms of action can aid in developing more effective treatments and prevention strategies, ultimately mitigating the impact of scorpion envenomation on public health in Brazil.

Purificación parcial de péptidos presentes en el veneno del escorpión Tityus macrochirus (Buthidae) y evaluación preliminar de su actividad citotóxica / Partial purification of peptides present in the Tityus macrochirus (Buthidae) scorpion venom and preliminary assessment of their cytotoxicity

RESUMEN Introducción. El veneno del escorpión posee péptidos con actividad neurotóxica que actúan principalmente en los canales iónicos del sistema nervioso de insectos y mamíferos. También se ha establecido su acción citolítica y anticancerígena, características biológicas que aún no se han explorado en el veneno del escorpión Tityus macrochirus. Objetivo. Evaluar si tanto el veneno total de T. macrochirus como la fracción de péptidos parcialmente purificados disminuyen el porcentaje de viabilidad de diferentes líneas celulares provenientes de tumores. Materiales y métodos. Mediante métodos cromatográficos, electroforéticos y de ultrafiltración con membranas de Amicon Ultra 0.5®, se identificaron y purificaron parcialmente los péptidos del veneno de T. macrochirus obtenido mediante estimulación eléctrica. Los ensayos de actividad citotóxica del veneno y de la fracción de péptidos se hicieron en líneas celulares provenientes de tumores con el método colorimétrico de reducción de la sal de tetrazolio (Mossman's Tetrazole Test, MTT). Resultados. El veneno de T. macrochirus posee péptidos con pesos moleculares entre 3 y 10 kDa, los cuales se purificaron parcialmente mediante ultrafiltración y se evaluaron mediante cromatografía líquida de alta resolución en fase inversa (Reverse Phase-High Pressure Liquid Chromatography, RP-HPLC). Los ensayos de citotoxicidad del veneno total de T. macrochirus evidenciaron una mayor disminución de la viabilidad en la línea celular PC3 que en las demás líneas celulares evaluadas, en tanto que la fracción parcialmente purificada de péptidos logró disminuir la viabilidad de la línea celular HeLa. Conclusión. Los péptidos del veneno de T. macrochirus presentaron actividad citotóxica en algunas de las líneas celulares provenientes de tumores, y se observó algún grado de selectividad frente a ellas.

ABSTRACT Introduction: Scorpion venom contains peptides with neurotoxic action primarily active on ion channels in the nervous system of insects and mammals. They are also characterized as cytolytic and anticancer, biological characteristics that have not yet been reported for the Tityus macrochirus venom. Objective: To assess if the total T. macrochirus venom and the fraction of partially purified peptides decrease the viability of various tumor-derived cell lines. Materials and methods: The scorpion venom was collected by electrical stimulation and, subsequently, subjected to chromatography, electrophoresis, and ultrafiltration with Amicon Ultra 0.5® membranes for the partial identification and purification of its peptides. The cytotoxic activity of the venom and the peptides fraction trials on tumor-derived cell lines were carried out by the MTT method. Results: The T. macrochirus scorpion venom has peptides with molecular weights ranging between 3 and 10 kDa. They were partially purified using the ultrafiltration technique, and assessed by the RP-HPLC method. Cytotoxicity trials with the whole T. macrochirus venom showed a higher viability decrease on the PC3 cell line compared to the other cell lines assessed, while the partially purified peptides decreased the HeLa cell line viability. Conclusion: Peptides in the T. macrochirus scorpion venom showed cytotoxic activity on some tumor-derived cell lines. We observed some degree of selectivity against other cell lines assessed.

Characterization of Leiurus abdullahbayrami (Scorpiones: Buthidae) venom: peptide profile, cytotoxicity and antimicrobial activity

Background Scorpion venoms are rich bioactive peptide libraries that offer promising molecules that may lead to the discovery and development of new drugs.Leiurus abdullahbayrami produces one of the most potent venoms among Turkish scorpions that provokes severe symptoms in envenomated victims.Methods In the present study, the peptide profile of the venom was investigated by electrophoretic methods, size-exclusion and reversed-phase chromatography and mass spectroscopy. Cytotoxic and antimicrobial effects were evaluated on a breast cancer cell line (MCF-7) and various bacterial and fungal species.Results Proteins make up approximately half of the dry weight of L. abdullahbayrami crude venom. Microfluidic capillary electrophoresis indicated the presence of 6 to 7 kDa peptides and proved to be a highly practical peptidomics tool with better resolution when compared to conventional polyacrylamide gel electrophoresis. Mass spectroscopy analysis helped us to identify 45 unique peptide masses between 1 to 7 kDa with a bimodal mass distribution peaking between molecular weights of 1 to 2 kDa (29%) and 3 to 4 kDa (31%). L. abdullahbayrami crude venom had a proliferative effect on MCF-7 cells, which may be explained by the high concentration of polyamines as well as potassium and calcium ions in the arachnid venoms. Antimicrobial effect was stronger on gram-negative bacteria.Conclusions This work represents the first peptidomic characterization of L. abdullahbayrami venom. Considering the molecular weight-function relationship of previously identified venom peptides, future bioactivity studies may lead to the discovery of novel potassium and chloride ion channel inhibitors as well as new antimicrobial peptides fromL. abdullahbayrami venom.

Construction of three different recombinant scorpion fusion proteins with bifunctional activity.

This is the first report of three different fusion proteins with an antitumor-analgesic peptide obtained from Chinese scorpion Buthus martensii Karsch (BmKAGAP). The fusion proteins were constructed in the form of chimeric toxins, aiming to obtain bifunctional analgesic and antitumor activity. The fusion proteins consisted of luteinizing hormone-releasing hormone (LHRH), three different types of flexible linkers (L1, Ser-Ser-His-His-His-His-His-His-Ser-Ser-Gly-Leu-Val-Pro-Arg-Gly-Ser-His-Met; L2, Gly-Gly-Gly-Ser-Gly-Gly-Gly-Ser; L3, Ser-Gly-Gly-Ser-Gly-Gly-Ser-Gly-Gly-Gly-Ser-Ser-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser), and BmKAGAP. The genes coding three fusion proteins were cloned and expressed in E. coli in soluble form. Following two successive column chromatographic separations, purified fusion proteins were obtained. These fusion proteins exhibited analgesic activity in mice and were cytotoxic to a hepatocellular carcinoma cell line Hep3B.

Hematological and biochemical profiles of rats submitted to experimental poisoning with Tityus serrulatus venom / Perfis hematológicos de ratos submetidos ao envenenamento escorpiônico experimental por Tityus serrulatus

The hematological and biochemical profiles of newly weaned rats submitted to experimental poisoning with T. serrulatus venom were evaluated. Fifteen recently weaned male Wistar rats (mean weight 130g) were distributed into three equal groups (n = 5). Animals in the control group (group A) received a subcutaneous injection of 400μL of ultra-pure water, while those in the experimental groups received, by identical route, 400μL of a solution containing 100μg (group B) or 450μg (group C) of scorpion venom dissolved in ultra-pure water. Red blood cells indexes, and differential leukocyte and total platelet counts were determined, together with levels of serum glucose, urea, creatinine, lactic dehydrogenase, aspartate aminotransferase, amylase, insulin, and cortisol. No significant differences between the control and experimental groups regarding red blood cells indexes were found. In contrast, significant increases (P<0.05) in neutrophils, lymphocytes, and monocytes were observed in animals from groups B and C compared with the control group, while the number of platelets decreased. Serum glucose concentration remained unchanged in all groups, but important alterations were observed in the values of urea and creatinine. The results show that scorpion venom was detrimental to renal function as demonstrated by the altered urea and creatinine levels. Pancreatic function was also impaired, as revealed by the increase in amylase activity and the reduction in insulin levels.

Avaliaram-se os perfis hematológico e bioquímico de ratos recém-desmamados submetidos ao envenenamento experimental com veneno de Tityus serrulatus. Quinze ratos Wistar machos, média de peso de 130g, foram distribuídos aleatoriamente em três grupos (n = 5). Os animais do grupo-controle A foram inoculados com 400μL de água ultrapura, os do grupo experimental B receberam 400μL de uma solução contendo 100μg de veneno e os do grupo experimental C receberam 400μL de uma solução contendo 450μg de veneno. Foram determinados os índices da série vermelha, a contagem total e diferencial dos leucócitos e a contagem total de plaquetas, bem como os níveis da desidrogenase lática, aspartato aminotransferase, amilase, glicose, ureia, creatinina, cortisol e insulina. Não houve diferenças significativas entre o grupo-controle e os experimentais com relação aos índices da série vermelha. Foram observados aumentos significativos (P<0,05) no número de neutrófilos, linfócitos e monócitos nos ratos dos grupos B e C, enquanto o número de plaquetas diminuiu. A concentração de glicose permaneceu inalterada em todos os grupos, mas foram observadas importantes alterações nos valores séricos de ureia e creatinina. Esses resultados mostraram que o veneno de escorpião comprometeu o funcionamento dos rins. Como demonstrado pelo aumento da atividade da amilase sérica e a redução dos níveis de insulina, a função pancreática também foi afetada.

The action of red scorpion (Mesobuthus tamulus coconsis, pocock) venom and its isolated protein fractions on blood sodium levels

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.(AU)

Effects of Androctonus crassicauda (Olivier, 1807) (scorpiones: buthidae) venom on rats: correlation among acetylcholinesterase activities and electrolytes levels

Scorpions can be considered living fossils because they have changed so little during the last 400 million years. They are venomous arthropods of the Arachnida class and regarded as relatives of spiders, ticks and mites. The aim of the present study was to evaluate the toxicity of Androctonus crassicauda (Olivier, 1807) venom and its effects on the acetylcholinesterase (AchE) activity and on electrolytes levels in rats. Animals were divided into seven groups of five rats each. Test groups received 250æg/kg of venom solution while control group was treated with 200æl of physiological saline solution (PSS). Blood samples were collected from the animals on the 1st, 2nd 4th, 8th, 12th, and 24th hours after subcutaneous injection of venom. Animals were monitored for 24 hours. Androctonus crassicauda venom significantly reduced AchE activity on the 12th hour when compared with control group. A statistically negative correlation between Na+ and K+ (p<0.05) and a positive correlation between Na+ and CL- (p<0.001) ions levels were observed after the administration of A. crassiccauda venom to rats. We can conclude that the differences in the electrolytes levels are due to acute renal failure, since elimination of toxin occurs primarily via the kidney.(AU)

Efectos del veneno de Buthus eupeus sobre la secreción gástrica / Effects of Buthus eupeus venom upon gastric secretion

La administración endovenosa del veneno del escorpión B. eupeus (a dosis de 0.25 mg/Kg) incrementa la secreción gastrica de acido y pepsinógeno en la rata, sin influenciar el volumen secretado. Ambas secreciones de acido y pepsinógeno fueron inhibidas por la atropina y la vagotomia. La secreción gastrica de ácido fue incrementada por el veneno de escorpion y este efecto fue disminuido por el tratamiento con cimetidina y hexamethonium. Los efectos del veneno de B. eupeus sobre la secreción gástrica de la rata son debidas a un efecto neurotoxico a nivel pre y postganglionar parasimpático

Brazilian scorpion venoms: pharmacological aspects

This review focuses on clinical, epidemiological and therapeutic aspects of envenomation caused by scorpions in Brazil. In addition, it presents studies with the crude venom and neurotoxins isolated from Tityus serrulatus scorpion venom. Such studies show the pharmacological effects of the venom and isolated toxins in experiments that use the scorpion venom and neurotoxins as neurobiological tools to study receptor sites in sodium channels and neurotransmitter release.