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1.
J. venom. anim. toxins incl. trop. dis ; 27: e20200180, 2021. tab, graf
Artigo em Inglês | VETINDEX, LILACS | ID: biblio-1287094

Resumo

Snake venoms are composed of pharmacologically active proteins that are evolutionarily diverse, stable and specific to targets. Hence, venoms have been explored as a source of bioactive molecules in treating numerous diseases. Recent evidences suggest that snake venom proteins may affect the formation of new blood vessels. Excessive angiogenesis has been implicated in several pathologies including tumours, diabetic retinopathy, arthritis, inter alia. In the present study, we have examined the effects of P-I metalloproteinases isolated from Bothrops moojeni (BmMP-1) and Bothrops atrox (BaMP-1) and L-amino acid oxidases (LAAO) isolated from B. moojeni (BmLAAO) and B. atrox (BaLAAO) on biochemical and functional aspects of angiogenesis. Methods: P-I metalloproteinases and LAAO were purified from venom by molecular size exclusion and ion-exchange chromatography and subsequently confirmed using mass spectrometry. The P-I metalloproteinases were characterized by azocaseinolytic, fibrinogenolytic and gelatinase activity and LAAO activity was assessed by enzyme activity on L-amino acids. Influence of these proteins on apoptosis and cell cycle in endothelial cells was analysed by flow cytometry. The angiogenic activity was determined by in vitro 3D spheroid assay, Matrigel tube forming assay, and in vivo agarose plug transformation in mice. Results: P-I metalloproteinases exhibited azocaseinolytic activity, cleaved α and partially β chain of fibrinogen, and displayed catalytic activity on gelatin. LAAO showed differential activity on L-amino acids. Flow cytometry analysis indicated that both P-I metalloproteinases and LAAO arrested the cells in G0/G1 phase and further induced both necrosis and apoptosis in endothelial cells. In vitro, P-I metalloproteinases and LAAO exhibited significant anti-angiogenic properties in 3D spheroid and Matrigel models by reducing sprout outgrowth and tube formation. Using agarose plug transplants in mice harbouring P-I metalloproteinases and LAAO we demonstrated a marked disruption of vasculature at the periphery. Conclusion: Our research suggests that P-I metalloproteinases and LAAO exhibit anti-angiogenic properties in vitro and in vivo.(AU)


Assuntos
Animais , Oxirredutases , Bothrops/fisiologia , Inibidores da Angiogênese , Venenos de Crotalídeos , Metaloproteases
2.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 27: e20200180, 2021. tab, graf
Artigo em Inglês | VETINDEX | ID: vti-31907

Resumo

Snake venoms are composed of pharmacologically active proteins that are evolutionarily diverse, stable and specific to targets. Hence, venoms have been explored as a source of bioactive molecules in treating numerous diseases. Recent evidences suggest that snake venom proteins may affect the formation of new blood vessels. Excessive angiogenesis has been implicated in several pathologies including tumours, diabetic retinopathy, arthritis, inter alia. In the present study, we have examined the effects of P-I metalloproteinases isolated from Bothrops moojeni (BmMP-1) and Bothrops atrox (BaMP-1) and L-amino acid oxidases (LAAO) isolated from B. moojeni (BmLAAO) and B. atrox (BaLAAO) on biochemical and functional aspects of angiogenesis. Methods: P-I metalloproteinases and LAAO were purified from venom by molecular size exclusion and ion-exchange chromatography and subsequently confirmed using mass spectrometry. The P-I metalloproteinases were characterized by azocaseinolytic, fibrinogenolytic and gelatinase activity and LAAO activity was assessed by enzyme activity on L-amino acids. Influence of these proteins on apoptosis and cell cycle in endothelial cells was analysed by flow cytometry. The angiogenic activity was determined by in vitro 3D spheroid assay, Matrigel tube forming assay, and in vivo agarose plug transformation in mice. Results: P-I metalloproteinases exhibited azocaseinolytic activity, cleaved α and partially β chain of fibrinogen, and displayed catalytic activity on gelatin. LAAO showed differential activity on L-amino acids. Flow cytometry analysis indicated that both P-I metalloproteinases and LAAO arrested the cells in G0/G1 phase and further induced both necrosis and apoptosis in endothelial cells. In vitro, P-I metalloproteinases and LAAO exhibited significant anti-angiogenic properties in 3D spheroid and Matrigel models by reducing sprout outgrowth and tube formation. Using agarose plug transplants in mice harbouring P-I metalloproteinases and LAAO we demonstrated a marked disruption of vasculature at the periphery. Conclusion: Our research suggests that P-I metalloproteinases and LAAO exhibit anti-angiogenic properties in vitro and in vivo.(AU)


Assuntos
Animais , Oxirredutases , Bothrops/fisiologia , Inibidores da Angiogênese , Venenos de Crotalídeos , Metaloproteases
3.
J. venom. anim. toxins incl. trop. dis ; 26: e20190058, 2020. tab, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1135137

Resumo

Lack of complete genomic data of Bothrops jararaca impedes molecular biology research focusing on biotechnological applications of venom gland components. Identification of full-length coding regions of genes is crucial for the correct molecular cloning design. Methods: RNA was extracted from the venom gland of one adult female specimen of Bothrops jararaca. Deep sequencing of the mRNA library was performed using Illumina NextSeq 500 platform. De novo assembly of B. jararaca transcriptome was done using Trinity. Annotation was performed using Blast2GO. All predicted proteins after clustering step were blasted against non-redundant protein database of NCBI using BLASTP. Metabolic pathways present in the transcriptome were annotated using the KAAS-KEGG Automatic Annotation Server. Toxins were identified in the B. jararaca predicted proteome using BLASTP against all protein sequences obtained from Animal Toxin Annotation Project from Uniprot KB/Swiss-Pro database. Figures and data visualization were performed using ggplot2 package in R language environment. Results: We described the in-depth transcriptome analysis of B. jararaca venom gland, in which 76,765 de novo assembled isoforms, 96,044 transcribed genes and 41,196 unique proteins were identified. The most abundant transcript was the zinc metalloproteinase-disintegrin-like jararhagin. Moreover, we identified 78 distinct functional classes of proteins, including toxins, inhibitors and tumor suppressors. Other venom proteins identified were the hemolytic lethal factors stonustoxin and verrucotoxin. Conclusion: It is believed that the application of deep sequencing to the analysis of snake venom transcriptomes may represent invaluable insight on their biotechnological potential focusing on candidate molecules.(AU)


Assuntos
Animais , Bothrops , Bothrops/fisiologia , Proteoma , Venenos de Crotalídeos , Perfilação da Expressão Gênica , Metaloproteases , Transcriptoma , Biologia Molecular , Análise por Conglomerados , Sequenciamento de Nucleotídeos em Larga Escala
4.
Cad. técn. Vet. Zoot. ; (75): 15-26, dez. 2014. ilus
Artigo em Português | VETINDEX | ID: vti-481331

Resumo

O gênero Bothrops continha mais de 60 espécies, distribuídas desde o norte do México até o sul da Argentina, mas várias espécies foram redistribuídas em outros sete gêneros. O Brasil possui a maioria das espécies remanescentes do gênero Bothrops, mas apenas duas espécies do gênero Bothriopsis (B. bilineata e B. taeniata) e duas de Bothrocophias (B. hyoprora e B. microphtalmus).


Assuntos
Animais , Serpentes/anatomia & histologia , Serpentes/fisiologia , Animais Peçonhentos/anatomia & histologia , Animais Peçonhentos/fisiologia , Bothrops/anatomia & histologia , Bothrops/fisiologia , Mordeduras de Serpentes/diagnóstico , Mordeduras de Serpentes/patologia , Peçonhas/análise , Sorologia , Peçonhas/efeitos adversos
5.
Cad. téc. vet. zootec ; (75): 15-26, dez. 2014. ilus
Artigo em Português | VETINDEX | ID: biblio-1471476

Resumo

O gênero Bothrops continha mais de 60 espécies, distribuídas desde o norte do México até o sul da Argentina, mas várias espécies foram redistribuídas em outros sete gêneros. O Brasil possui a maioria das espécies remanescentes do gênero Bothrops, mas apenas duas espécies do gênero Bothriopsis (B. bilineata e B. taeniata) e duas de Bothrocophias (B. hyoprora e B. microphtalmus).


Assuntos
Animais , Animais Peçonhentos/anatomia & histologia , Animais Peçonhentos/fisiologia , Bothrops/anatomia & histologia , Bothrops/fisiologia , Mordeduras de Serpentes/diagnóstico , Mordeduras de Serpentes/patologia , Serpentes/anatomia & histologia , Serpentes/fisiologia , Peçonhas/análise , Peçonhas/efeitos adversos , Sorologia
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