RESUMO
BACKGROUND: Almost all Tityus characterized toxins are from subgenera Atreus and Tityus, there are only a few data about toxins produced by Archaeotityus, an ancient group in Tityus genus. METHODS: Tityus (Archaeotityus) mattogrossensis crude venom was fractionated by high performance liquid chromatography, the major fractions were tested in a frog sciatic nerve single sucrose-gap technique. Two fractions (Tm1 and Tm2) were isolated, partially sequenced by MALDI-TOF/MS and electrophysiological assayed on HEK293 Nav 1.3, HEK293 Nav 1.6, DUM and DRG cells. RESULTS: The sucrose-gap technique showed neurotoxicity in four fractions. One fraction caused a delay of action potential repolarization and other three caused a reduction in amplitude. An electrophysiological assay showed that Tm1 is active on HEK293 Nav 1.3, HEK293 Nav 1.6, DUM and DRG cells, and Tm2 on HEK293 Nav 1.3 and DRG cells, but not in HEK293 Nav 1.6. In addition, Tm1 and Tm2 did promote a shift to more negative potentials strongly suggesting that both are α-NaScTx. CONCLUSION: Although Tityus (Archaeotityus) mattogrossensis is considered an ancient group in Tityus genus, the primary structure of Tm1 and Tm2 is more related to Tityus subgenus. The patch clamp electrophysiological tests suggest that Tm1 and Tm2 are NaScTx, and also promoted no shift to more negative potentials, strongly suggesting that both are α-NaScTx. This paper aimed to explore and characterize for the first time toxins from the ancient scorpion Tityus (Archaeotityus) mattogrossensis.
RESUMO
Abstract Background: Almost all Tityus characterized toxins are from subgenera Atreus and Tityus, there are only a few data about toxins produced by Archaeotityus, an ancient group in Tityus genus. Methods: Tityus (Archaeotityus) mattogrossensis crude venom was fractionated by high performance liquid chromatography, the major fractions were tested in a frog sciatic nerve single sucrose-gap technique. Two fractions (Tm1 and Tm2) were isolated, partially sequenced by MALDI-TOF/MS and electrophysiological assayed on HEK293 Nav 1.3, HEK293 Nav 1.6, DUM and DRG cells. Results: The sucrose-gap technique showed neurotoxicity in four fractions. One fraction caused a delay of action potential repolarization and other three caused a reduction in amplitude. An electrophysiological assay showed that Tm1 is active on HEK293 Nav 1.3, HEK293 Nav 1.6, DUM and DRG cells, and Tm2 on HEK293 Nav 1.3 and DRG cells, but not in HEK293 Nav 1.6. In addition, Tm1 and Tm2 did promote a shift to more negative potentials strongly suggesting that both are -NaScTx. Conclusion: Although Tityus (Archaeotityus) mattogrossensis is considered an ancient group in Tityus genus, the primary structure of Tm1 and Tm2 is more related to Tityus subgenus. The patch clamp electrophysiological tests suggest that Tm1 and Tm2 are NaScTx, and also promoted no shift to more negative potentials, strongly suggesting that both are -NaScTx. This paper aimed to explore and characterize for the first time toxins from the ancient scorpion Tityus (Archaeotityus) mattogrossensis.
RESUMO
Background: Almost all Tityus characterized toxins are from subgenera Atreus and Tityus, there are only a few data about toxins produced by Archaeotityus, an ancient group in Tityus genus. Methods: Tityus (Archaeotityus) mattogrossensis crude venom was fractionated by high performance liquid chromatography, the major fractions were tested in a frog sciatic nerve single sucrose-gap technique. Two fractions (Tm1 and Tm2) were isolated, partially sequenced by MALDI-TOF/MS and electrophysiological assayed on HEK293 Nav 1.3, HEK293 Nav 1.6, DUM and DRG cells. Results: The sucrose-gap technique showed neurotoxicity in four fractions. One fraction caused a delay of action potential repolarization and other three caused a reduction in amplitude. An electrophysiological assay showed that Tm1 is active on HEK293 Nav 1.3, HEK293 Nav 1.6, DUM and DRG cells, and Tm2 on HEK293 Nav 1.3 and DRG cells, but not in HEK293 Nav 1.6. In addition, Tm1 and Tm2 did promote a shift to more negative potentials strongly suggesting that both are α-NaScTx. Conclusion: Although Tityus (Archaeotityus) mattogrossensis is considered an ancient group in Tityus genus, the primary structure of Tm1 and Tm2 is more related to Tityus subgenus. The patch clamp electrophysiological tests suggest that Tm1 and Tm2 are NaScTx, and also promoted no shift to more negative potentials, strongly suggesting that both are α-NaScTx. This paper aimed to explore and characterize for the first time toxins from the ancient scorpion Tityus (Archaeotityus) mattogrossensis.(AU)
Assuntos
Animais , Venenos de Escorpião/isolamento & purificação , Venenos de Escorpião/classificação , Cromatografia Líquida/métodos , Sacarose/análise , Fenômenos Eletrofisiológicos/fisiologiaRESUMO
Objetivo Avaliar o uso da espectroscopia Raman como possível instrumento para a detecção do Ibuprofeno em diferentes formulações farmacêuticas a fim de se propor uma metodologia a ser utilizada em processos de controle de qualidade na produção destes medicamentos. Métodos 05 amostras de cada grupo do medicamento (referência, genérico e similar) concentrado em 100 mg/dL, foram submetidas a espectroscopia Raman (espectrômetro dispersivo com 830 nm e 300 mW de excitação acoplado a um cabo de fibras ópticas Raman probe) e as aquisições foram replicadas 10 vezes de 3 segundos, com exposição total de 30 s para a coleta de cada espectro. Resultados Ao se comparar o medicamento similar com o de referência, percebem-se picos mais elevados e largos no similar, principalmente nos deslocamentos de 510, 660 e 884 cm1. Já os espectros dos medicamentos de referência e genérico mostraram-se mais semelhantes entre si. Conclusão As três apresentações comerciais contêm o princípio ativo Ibuprofeno, porém os picos em diferentes intensidades sugerem diferentes concentrações nas formulações avaliadas.
Objective To evaluate the use of Raman spectroscopy as a possible instrument for the detection of ibuprofen in different pharmaceutical formulations in order to propose a methodology to be used in quality control processes in the production of these drugs. Methods 05 samples from each drug group (reference, 100 mg / dL, were subjected to Raman spectroscopy (830 nm dispersive spectrometer and 300 mW excitation coupled to a Raman probe fiber optic cable) and acquisitions replicated 10 times 3 seconds with exposure 30 s total for the collection of each spectrum. Results When comparing the similar drug with the reference drug, higher and broader peaks in the similar one can be noticed, especially in the displacements of 510, 660 and 884 cm1. Already the spectra of reference and generic drugs were more similar to each other. Conclusion The three commercial presentations contain the active ingredient Ibuprofen, but the peaks at different intensities suggest different concentrations in the formulations evaluated.