Essential Oil from Lippia origanoides (Verbenaceae): Haemostasis and Enzymes Activity Alterations

Background: The search for natural inhibitors of snake venom toxins is essential to supplement or even replace the serum therapy. The aim of this work was to evaluate the pharmacological properties of essential oil from Lippia origanoides Kunth. (Verbenaceae). Methods: The oil was extracted by hydrodistillation and the constituents were identified and quantified by GC-MS and GC-FID. The essential oil from L. origanoides was evaluated in hemolysis tests, on the activities of phospholipases A2 and serine proteases and in coagulation and thrombolysis induced by different snake venoms. Results: The major constituents of essential oil were carvacrol, p-cymene, γ-terpinene, and thymol. The oil inhibited approximately 10 % of the phospholipase A2 activity induced by Bothrops atrox, Bothrops jararaca, Bothrops jararacussu and Bothrops moojeni venoms and was not cytotoxic against erythrocytes. However, previous incubation of the oil with B. jararacussu, B. moojeni, and Crotalus durissus terrificus (C.d.t.) venoms resulted in potentiation of hemolytic activity (30 % and 50 % for 0.6 µL mL-1 and 1.2 µL mL-1, respectively). The essential oil presented a procoagulant effect on human citrated plasma, potentiated the thrombolytic action of proteases and phospholipases A2 present in B. jararacussu venom, and serine protease activity induced by B. jararaca and Lachesis muta venoms. When pre-incubated with the C.d.t. venom, however, prothrombotic activity was observed. Conclusion: The results obtained in this work amplify the pharmacological characterization of the essential oil from L. origanoides. However, new studies are fundamental to define the action mechanisms and determine pharmaceutical applications.

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Caryospora bigenetica (Apicomplexa: Eimeriidae) in South America: new hosts and distribution records

Revista Brasileira de Parasitologia Veterinária ◽

10.1590/s1984-29612015002 ◽

2015 ◽

Vol 24 (1) ◽

pp. 101-104 ◽

Cited By ~ 1

Author(s):

Lúcio André Viana ◽

Kamilla Costa Mecchi ◽

Leonardo França do Nascimento ◽

Heitor Miraglia Herrera ◽

Paula Helena Santa-Rita ◽

...

Keyword(s):

South America ◽

Polar Granule ◽

First Record ◽

Stieda Body ◽

Mato Grosso ◽

Sporocyst Residuum ◽

The Family ◽

New Hosts ◽

Crotalus Durissus ◽

Bothrops Moojeni

The coccidian Caryospora bigenetica was first described in the snake Crotalus horridus (Viperidae) from United States of America. This study represents the first record of the occurrence of C. bigenetica in snakes in South America. Feces were sampled between November 2013 and May 2014 from 256 wild snakes maintained in scientific breeding facilities in the states of Mato Grosso do Sul (MS; n = 214) and Rio de Janeiro (RJ; n = 42), Brazil. Caryospora bigenetica was found in 14 (5.6%) snakes, all belonging to the family Viperidae. Ten Bothrops moojeni and two Crotalus durissus from MS were infected. The coccidian was also found in one C. durissus and in one Bothrops jararacussu from the state of RJ. The oocysts were spherical with a double wall, the exterior lightly mammillated, striations apparent in transverse view, 13.0 µm (12 – 14); polar granule fixed in the internal wall. Sporocysts oval or pyriform, 10.0 × 8.0 µm (9 – 11 × 8 – 9); Stieda body discoid; sub-Stieda body present; sporocyst residuum present, formed by a group of spheroid bodies between sporozoites. This study increases the number of viperid hosts of C. bigenetica and expands the geographical distribution to South America.

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Studies on the specificity of CNF, a phospholipase A2 inhibitor isolated from the blood plasma of the South American rattlesnake (Crotalus durissus terrificus). I. Interaction with PLA2 from Lachesis muta muta snake venom

Toxicon ◽

10.1016/s0041-0101(99)00116-6 ◽

1999 ◽

Vol 37 (12) ◽

pp. 1747-1759 ◽

Cited By ~ 24

Author(s):

C.L. Fortes-Dias ◽

M.L.D. Jannotti ◽

F.J.L. Franco ◽

A. Magalhães ◽

C.R. Diniz

Keyword(s):

Phospholipase A2 ◽

Blood Plasma ◽

Snake Venom ◽

South American ◽

The South ◽

Crotalus Durissus Terrificus ◽

Phospholipase A2 Inhibitor ◽

Lachesis Muta ◽

Crotalus Durissus

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Edema Induced by a Crotalus durissus terrificus Venom Serine Protease (Cdtsp 2) Involves the PAR Pathway and PKC and PLC Activation

International Journal of Molecular Sciences ◽

10.3390/ijms19082405 ◽

2018 ◽

Vol 19 (8) ◽

pp. 2405 ◽

Cited By ~ 1

Author(s):

Caroline Costa ◽

Mariana Belchor ◽

Caroline Rodrigues ◽

Daniela Toyama ◽

Marcos de Oliveira ◽

...

Keyword(s):

Serine Protease ◽

Snake Venom ◽

Serine Proteases ◽

Three Dimensional ◽

Negative Control ◽

Protease Activated Receptors ◽

Crotalus Durissus Terrificus ◽

Three Dimensional Modeling ◽

Cox 2 ◽

Crotalus Durissus

Snake venom serine proteases (SVSPs) represent an essential group of enzymatic toxins involved in several pathophysiological effects on blood homeostasis. Some findings suggest the involvement of this class of enzymatic toxins in inflammation. In this paper, we purified and isolated a new gyroxin isoform from the Crotalus durissus terrificus (Cdt) venom, designated as Cdtsp 2, which showed significant proinflammatory effects in a murine model. In addition, we performed several studies to elucidate the main pathway underlying the edematogenic effect induced by Cdtsp 2. Enzymatic assays and structural analysis (primary structure analysis and three-dimensional modeling) were closely performed with pharmacological assays. The determination of edematogenic activity was performed using Cdtsp 2 isolated from snake venom, and was applied to mice treated with protein kinase C (PKC) inhibitor, phospholipase C (PLC) inhibitor, dexamethasone (Dexa), antagonists for protease-activated receptors (PARs), or saline (negative control). Additionally, we measured the levels of cyclooxygenase 2 (COX-2), malondialdehyde (MDA), and prostaglandin E2 (PGE2). Cdtsp 2 is characterized by an approximate molecular mass of 27 kDa, an isoelectric point (pI) of 4.5, and significant fibrinolytic activity, as well as the ability to hydrolyze Nα-benzoyl-l-arginine 4-nitroanilide (BAPNA). Its primary and three-dimensional structures revealed Cdtsp 2 as a typical snake venom serine protease that induces significant edema via the metabolism of arachidonic acid (AA), involving PARs, PKC, PLC, and COX-2 receptors, as well as inducing a significant increase in MDA levels. Our results showed that Cdtsp 2 is a serine protease with significant enzymatic activity, and it may be involved in the degradation of PAR1 and PAR2, which activate PLC and PKC to mobilize AA, while increasing oxidative stress. In this article, we provide a new perspective for the role of SVSPs beyond their effects on blood homeostasis.

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Bothrops moojeni and Crotalus durissus terrificus whole venoms activate transcription factor NF-κB in 3T3-L1 preadipocytes

Toxicon ◽

10.1016/j.toxicon.2019.06.092 ◽

2019 ◽

Vol 168 ◽

pp. S20

Author(s):

Rodrigo Maia Marques ◽

Priscila Signor Motta ◽

Danilo Dos Santos Teixeira ◽

Elbio Leiguez ◽

Catarina Teixeira

Keyword(s):

Transcription Factor ◽

Crotalus Durissus Terrificus ◽

Activate Transcription Factor ◽

Crotalus Durissus ◽

Bothrops Moojeni

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Study of the Proteolytic Activity of the Tropical Legume Crotalaria spectabilis

Zeitschrift für Naturforschung C ◽

10.1515/znc-2012-9-1008 ◽

2012 ◽

Vol 67 (9-10) ◽

pp. 495-509 ◽

Cited By ~ 1

Author(s):

Juliana da Silva Pacheco ◽

Raquel Elisa da Silva-Lopez

Keyword(s):

Proteolytic Activity ◽

Serine Proteases ◽

Ph Value ◽

Cysteine Proteases ◽

Leaf Extracts ◽

Serine Protease Activity ◽

Proteolytic Activities ◽

Low Levels ◽

Crotalaria Spectabilis

The characterization of legume proteases contributes to the understanding of the physiology of plants and their interaction with the environment. Thirteen extracts from various parts of Crotalaria spectabilis were made using different extraction systems. The highest protein content was found in seeds, and the most pronounced proteolytic activity was observed in leaf extracts, with an optimal pH value in the alkaline range. Proteases in extracts from roots, stems, and fl owers were active in various pH ranges. Proteases in all extracts were maximally active between 30 °C and 60 °C and were thermostable (24 h, 60 °C). Hemoglobin, bovine serum albumin, casein, and gelatin were hydrolyzed by C. spectabilis extracts in different ways. The highest serine protease activity was found in leaves. Seeds contained high levels of serine proteases and low levels of cysteine proteases. Flowers, roots, and stems contained different levels of serine, aspartic, and metalloproteases, respectively. The proteolytic activities in extracts were modulated by cations and oxidants to various degrees. C. spectabilis proteases are differentially expressed in distinctive organs, and their stability against heat and oxidants makes this plant an important source of stable proteases

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Multifaceted Analyses of Epidermal Serine Protease Activity in Patients with Atopic Dermatitis

International Journal of Molecular Sciences ◽

10.3390/ijms21030913 ◽

2020 ◽

Vol 21 (3) ◽

pp. 913 ◽

Cited By ~ 1

Author(s):

Hayato Nomura ◽

Mutsumi Suganuma ◽

Takuya Takeichi ◽

Michihiro Kono ◽

Yuki Isokane ◽

...

Keyword(s):

Atopic Dermatitis ◽

Serine Protease ◽

Protease Activity ◽

Serine Proteases ◽

Immunoglobulin E ◽

Total Serum ◽

Tape Stripping ◽

Blood Eosinophil ◽

Peripheral Blood Eosinophil ◽

Serine Protease Activity

The serine proteases kallikrein-related peptidase (KLK) 5 and KLK7 cleave cell adhesion molecules in the epidermis. Aberrant epidermal serine protease activity is thought to play an important role in the pathogenesis of atopic dermatitis (AD). We collected the stratum corneum (SC) from healthy individuals (n = 46) and AD patients (n = 63) by tape stripping and then measuring the trypsin- and chymotrypsin-like serine protease activity. We also analyzed the p.D386N and p.E420K of SPINK5 variants and loss-of-function mutations of FLG in the AD patients. The serine protease activity in the SC was increased not only in AD lesions but also in non-lesions of AD patients. We found, generally, that there was a positive correlation between the serine protease activity in the SC and the total serum immunoglobulin E (IgE) levels, serum thymus and activation-regulated chemokine (TARC) levels, and peripheral blood eosinophil counts. Moreover, the p.D386N or p.E420K in SPINK5 and FLG mutations were not significantly associated with the SC’s serine protease activity. Epidermal serine protease activity was increased even in non-lesions of AD patients. Such activity was found to correlate with a number of biomarkers of AD. Further investigations of serine proteases might provide new treatments and prophylaxis for AD.

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Venomics of Trimeresurus (Popeia) nebularis, the Cameron Highlands Pit Viper from Malaysia: Insights into Venom Proteome, Toxicity and Neutralization of Antivenom

Toxins ◽

10.3390/toxins11020095 ◽

2019 ◽

Vol 11 (2) ◽

pp. 95 ◽

Cited By ~ 12

Author(s):

Choo Tan ◽

Kae Tan ◽

Tzu Ng ◽

Evan Quah ◽

Ahmad Ismail ◽

...

Keyword(s):

Snake Venom ◽

Serine Proteases ◽

Cross Reactivity ◽

Secretory Proteins ◽

Proteomic Profiling ◽

Phospholipases A2 ◽

Central Highland ◽

Liquid Chromatography Tandem Mass ◽

Cameron Highlands ◽

Venom Proteins

Trimeresurus nebularis is a montane pit viper that causes bites and envenomation to various communities in the central highland region of Malaysia, in particular Cameron’s Highlands. To unravel the venom composition of this species, the venom proteins were digested by trypsin and subjected to nano-liquid chromatography-tandem mass spectrometry (LC-MS/MS) for proteomic profiling. Snake venom metalloproteinases (SVMP) dominated the venom proteome by 48.42% of total venom proteins, with a characteristic distribution of P-III: P-II classes in a ratio of 2:1, while P-I class was undetected. Snaclecs constituted the second most venomous protein family (19.43%), followed by snake venom serine proteases (SVSP, 14.27%), phospholipases A2 (5.40%), disintegrins (5.26%) and minor proteins including cysteine-rich secretory proteins, L-amino acid oxidases, phosphodiesterases, 5′-nucleotidases. The venomic profile correlates with local (painful progressive edema) and systemic (hemorrhage, coagulopathy, thrombocytopenia) manifestation of T. nebularis envenoming. As specific antivenom is unavailable for T. nebularis, the hetero-specific Thai Green Pit viper Monovalent Antivenom (GPVAV) was examined for immunological cross-reactivity. GPVAV exhibited good immunoreactivity to T. nebularis venom and the antivenom effectively cross-neutralized the hemotoxic and lethal effects of T. nebularis (lethality neutralizing potency = 1.6 mg venom per mL antivenom). The findings supported GPVAV use in treating T. nebularis envenoming.

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O CONHECIMENTO TRADICIONAL SOBRE AS SERPENTES EM UMA COMUNIDADE RIBEIRINHA NO CENTRO-LESTE DA AMAZÔNIA

Ethnoscientia - Brazilian Journal of Ethnobiology and Ethnoecology ◽

10.22276/ethnoscientia.v3i0.157 ◽

2018 ◽

Vol 3 ◽

Cited By ~ 1

Author(s):

Lourival Baía Vasconcelos Neto ◽

Hipócrates De Menezes Chalkidis ◽

Ilka Andreyce Da Silva Brito ◽

Antonio Samuel Garcia da Silva

Keyword(s):

Lachesis Muta ◽

Crotalus Durissus

A etnociência investiga a origem das atitudes e relações humanas com o ambiente. Nesse contexto, a etnozoologia estuda o conhecimento tradicional do homem sobre os animais. O presente trabalho tem como objetivo obter informações sobre o conhecimento tradicional dos moradores da comunidade de Paracari (Monte Alegre, Pará, Brasil) acerca das serpentes, questionando sobre como eles denominam, caracterizam e lidam com esses animais e com os possíveis acidentes. Para isso, durante três campanhas efetuadas nos meses de novembro e dezembro de 2015, questionários contendo tópicos previamente selecionados foram aplicados a 42 moradores selecionados aleatoriamente e que se dispuseram a contribuir. Destes, 95,23% alegaram já terem tido contato com serpentes e, com relação à hostilidade, 59,53% afirmaram matar a serpente independente da espécie. Quanto à ocorrência de acidentes ofídicos no ciclo familiar, 61,90% dos entrevistados confirmaram tal ocorrência, mas apenas 38,46% procuraram atendimento médico. O tratamento de origem popular mais comumente mencionado no caso de picada de cobra foi um extrato de planta artesanal chamado "Pau-X", fabricado, engarrafado e vendido livremente na região. Apenas Eunectes murinus, Lachesis muta e Crotalus durissus foram corretamente identificadas e caracterizadas pelos moradores. Algumas espécies foram identificadas corretamente, mas, em várias ocasiões, foram caracterizadas incorretamente como venenosas ou não. Todos os entrevistados confirmaram nunca ter tido acesso a informações ou participado de práticas educativo-ambientais sobre serpentes, o que pode contribuir diretamente para o uso de práticas inadequadas na identificação das espécies e no tratamento de acidentes ofídicos.

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O CONHECIMENTO TRADICIONAL SOBRE AS SERPENTES EM UMA COMUNIDADE RIBEIRINHA NO CENTRO-LESTE DA AMAZÔNIA

Ethnoscientia - Brazilian Journal of Ethnobiology and Ethnoecology ◽

10.18542/ethnoscientia.v3i0.10206 ◽

2018 ◽

Vol 3 ◽

Author(s):

Lourival Baía Vasconcelos Neto ◽

Hipócrates De Menezes Chalkidis ◽

Ilka Andreyce Da Silva Brito ◽

Antonio Samuel Garcia da Silva

Keyword(s):

Lachesis Muta ◽

Crotalus Durissus

A etnociência investiga a origem das atitudes e relações humanas com o ambiente. Nesse contexto, a etnozoologia estuda o conhecimento tradicional do homem sobre os animais. O presente trabalho tem como objetivo obter informações sobre o conhecimento tradicional dos moradores da comunidade de Paracari (Monte Alegre, Pará, Brasil) acerca das serpentes, questionando sobre como eles denominam, caracterizam e lidam com esses animais e com os possíveis acidentes. Para isso, durante três campanhas efetuadas nos meses de novembro e dezembro de 2015, questionários contendo tópicos previamente selecionados foram aplicados a 42 moradores selecionados aleatoriamente e que se dispuseram a contribuir. Destes, 95,23% alegaram já terem tido contato com serpentes e, com relação à hostilidade, 59,53% afirmaram matar a serpente independente da espécie. Quanto à ocorrência de acidentes ofídicos no ciclo familiar, 61,90% dos entrevistados confirmaram tal ocorrência, mas apenas 38,46% procuraram atendimento médico. O tratamento de origem popular mais comumente mencionado no caso de picada de cobra foi um extrato de planta artesanal chamado "Pau-X", fabricado, engarrafado e vendido livremente na região. Apenas Eunectes murinus, Lachesis muta e Crotalus durissus foram corretamente identificadas e caracterizadas pelos moradores. Algumas espécies foram identificadas corretamente, mas, em várias ocasiões, foram caracterizadas incorretamente como venenosas ou não. Todos os entrevistados confirmaram nunca ter tido acesso a informações ou participado de práticas educativo-ambientais sobre serpentes, o que pode contribuir diretamente para o uso de práticas inadequadas na identificação das espécies e no tratamento de acidentes ofídicos.

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