Thrombolysis, clotting, and genotoxic activities modulated by essential oils extracted from Lippia alba

Abstract The composition and pharmacological properties of Lippia alba (Mill.) (L. alba) (Verbenaceae) flower and leaf essential oils (EO) were determined in this study. The major constituents in the flower EO were geranial (49.83%) and neral (32.75%), and in the leaf EO were geranial (38.06%), neral (31.02%), and limonene (18.03%). Flower EO inhibited thrombolysis induced by Bothrops moojeni (B. moojeni) and Lachesis muta muta (L. muta muta) venoms (0.05-1.2 µL mL-1). When tested against L. muta muta venom, the protective effect was smaller in both EO. The EOs prolonged the clotting time induced by L. muta muta venom and a procoagulant effect was observed on B. moojeni. In the comet assay, the flower EO presented anti-genotoxic action (damage frequency of only 11.6 - 34.9%) against the L. muta muta venom. The positive control (Doxorubicin) and the venom alone presented a damage frequency of 80.3% and 70.7%, respectively. The flower EO protected DNA from damage induced by L. muta muta venom. L. alba leaf and flower EOs presented anti-genotoxic action

Dendrotoxin-kappa suppresses tumor growth induced by human lung adenocarcinoma A549 cells in nude mice

Voltage-gated K+ (Kv) channels have been considered to be a regulator of membrane potential and neuronal excitability. Recently, accumulated evidence has indicated that several Kv channel subtypes contribute to the control of cell proliferation in various types of cells and are worth noting as potential emerging molecular targets of cancer therapy. In the present study, we investigated the effects of the Kv1.1-specific blocker, dendrotoxin-kappa (DTX-kappa), on tumor formation induced by the human lung adenocarcinoma cell line A549 in a xenograft model. Kv1.1 mRNA and protein was expressed in A549 cells and the blockade of Kv1.1 by DTX-kappa, reduced tumor formation in nude mice. Furthermore, treatment with DTX-kappa significantly increased protein expression of p21Waf1/Cip1, p27Kip1, and p15INK4B and significantly decreased protein expression of cyclin D3 in tumor tissues compared to the control. These results suggest that DTX-kappa has anti-tumor effects in A549 cells through the pathway governing G1-S transition.

blocking activity of different toxins against potassium channels Kv3.4 in RLE cells

K[+] channel toxins are essential tools for the first purifications, analysis of subunit structures and brain localization of voltage-gated K[+] [Kv] channels. The effects of a lot of toxins on Kv are not fully known. Using whole-cell patch clamping technique the action of a series of toxins on Kv3.4 current in rat liver cells with expressed Kv3.4 channels [RLE] cloned cells was investigated. The cells were grown in Williams E medium and after 6-8 days, they were suitable for patch clamping. A family of currents was recorded during voltage-clamp steps to various potentials applied from a holding potential of-60 mV to 60-80 mV in 10 mV increments. Upon depolarization, all channels were opened with a sigmoidal time course, reached to the peak within a few 10[th] of milliseconds and then slowly inactivated. Bath application of tetraethyl ammonium [TEA] or 3, 4-diaminopyridine [DAP] reduced the current dose dependently and inhibited it completely at 3 mM and 25 micro M respectively. The Bunodosoma granulifera [BgK] and Heteractis magniflca [HmK] toxins at concentrations up to 30 and 10 micro M respectively could not completely inhibit the current. On the hand, toxins such as beta-bungarotoxin, corotoxin, novel toxin and dendrotoxins I [DIP] and K [DPK] even in high concentrations [up to 100 mM] had not any significant effect on Kv3.4 current. Comparison of chemical structures of these effective agents with other reported effective toxins such as blood depressing substances [BDS 1 and II] show no homology between them, but specially the potency of 3, 4-DAP is comparable with these toxins. These results showed that, the Kv3.4 is more sensitive than other K[+] channels to 3, 4-DAP. The sensitivity of this channel to the TEA is low [at mM concentration]. More investigation is necessary to find more selective and potent inhibitor of Kv3.4 channels

Diagnóstico diferencial de Naja con otros venenos de víbora / Diferencial diagnostic of Naja with other snake venoms

La idea de este trabajo es establecer un diagnóstico diferencial de Naja con otros medicamentos pero poniendo el acento en su cuadro cardíaco donde la rapidez diagnóstica es imprescindible

Actividad hemolítica de venenos de serpientes de los géneros Bothrops, Lachesis, Crotalus y Micrurus (Serpentes: Viperidae y Elapidae) / Haemolytic activity of venoms from snakes of the genera Bothrop, Lachesis, Crotalus and Micrurus (Serpentes: Viperidae and Elapidae)

Hemolytic activity of eight Peruvian snake venoms from the families Viperidae and Elapidae (Bothrops atrox, B. pictus, B. hyoprorus, B. bilineatus, B. neuwedii, Lachesis m. muta, Crotalus d. terrificus, Microrus tschudi), and three Brazilian viperids (B. jararacussu, B. alternatus and C. d. collilineatus) is described. None of the venoms cause direct lysis on washed human erythrocytes. However, all of then caused indirect hemolysis provided that the incubation medium contains an exogenous source of lecithin. Venom of Micrurus tschudi was the most hemolytic (HD50 2.8 ug/ml) while that of B. bilineatus was the least (HD50 681.3 ug/ml). Only six of eleven venoms showed parallel curves of hemolytic activity, and the HD50 varied from 198 to 681 ug/ml and the following decreasing order of hemolytic activity was obtained: L. muta, C. d. terrificus, C. d. collilineatus, B. hyoprorus, B. bilineatus, B. alternatus

Biochemical and behavioral effects of intraseptal microinjection of fasciculin, an irreversible acetylcholinesterase inhibitor

We examined the effect, in rats, of an intraseptal microinjection of fasciculin (FAS), an irreversible peptide acetylcholinesterase (AChE) inhibitor, on a)AChE activity measured in septum and hippocampus, b)3H-quinuclidiny benzylate (3H-QNB) and 3H-oxotremorine (3H-OXO) binding to hippocampal cholinergic muscarinic receptors, c) 3H-flunitrazepan (3H-FNZ) binding to hippocampal benzodiazepine receptors as a control for QNB and OXO binding, d) acquisition and retention in three different behavioral paradigms, i. e., water-finding (in which there is concomitant habituation to be apparatus), step-down inhibitory avoidance, and shuttle avoidance. AChE activity in septum decreased 2 days (-66%) and 5 days (-48%) after FAS microinjection; a slight reduction (-35%) occurred in the dorsal hippocampus on day 2 (P<0.05; N=6 per group); no changes in AChE activity were observed in ventral hippocampus ion day 2 or day 5. No changes in 3H-QNB, 3H-OXO, or 3H-FNZ binding constants were demonstrable in the hippocampus either 2 or 5 days after intraseptal FAS adminstration. No changes in training or test session performance in any of the three behavioral situations were observed 2-3 days after the intraseptal microinjection of FAS. The persistent inhibition of septal AChE caused by FAS microinjection into the septum is not sufficient to induce major changes either in hippocampal cholinergic muscarinic receptors, or in the learning or retention of behaviors regulated by the septum and/or hippocampus

Biochemical studies of liver & liver microsomes in envenomated rats.

Some biochemical parameters of liver and liver microsomes were studied in albino rats following administration of cobra and viper venoms at dose of 2 mg/kg body weight. The total protein content in cobra venom treated (CVT) animals and DNA and RNA contents of liver and liver microsomes were almost unaltered in both the venom treated animals while total protein content was significantly reduced in viper venom treated (VVT) animals. Alkaline and acid phosphatases activities of whole liver showed significant increase in both the venom treated animals whereas the rise in cholinesterase activity in CVT animals was not significant. Lactic acid content was significantly higher in CVT animals compared to either VVT animals or controls. The glycolytic enzymes viz., aldolase, phosphohexose isomerase and lactate dehydrogenase measured in hepatic microsomal fraction were significantly reduced while alanine and aspartate aminotransferases and gamma-glutamyl transpeptidase activities of liver microsomes were significantly elevated in both the venom treated animals compared to controls.

Changes in lipid profiles of brain of albino rats following envenomation.

Effect of high doses of cobra venom (150 micrograms/120 +/- 20 g body weight) and viper venom (300 micrograms/120 +/- 20 g body weight) on total lipid, triglyceride, phospholipid, cholesterol, high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) of brain of albino rats was studied. Total lipid (TL) triglyceride (TG) and phospholipid (PL) are decreased in both viper and cobra venom treated groups while cholesterol (C), and LDL-C are increased in both the groups in relation to controlled ones. HDL-C content was almost unaltered. Decrease in triglyceride and phospholipid may be due to effect of lipases and phospholipases whereas increased cholesterol and LDL-C may be attributed to lysis of cell membrane.

Acidentes por serpentes peçonhentas do Brasil / Accidents by snake venoms of Brazil

O presente artigo revisa conhecimentos referentes à incidência, epidemiologia, fisiopatologia, quadro clínico, diagnóstico e prognóstico dos acidentes por serpentes peçonhentas do Brasil que säo responsáveis por mais de 20.000 acidentes e 100 óbitos por ano. Pertencem a quatro gêneros: Bothrops, Crotalus, Lachesis e Micrurus. Executando as Micrurus, todas causam distúrbios na coagulaçäo sanguínea; as Lachesis e Bothrops provocam destruiçäo tecidual na regiäo da picada, as Crotalus e Micrurus, bloqueio na placa motora e somente as Crotalus, miotoxicidade sistêmica. O quadro clínico geralmente é suficiente para o diagnóstico, exceto na distinçäo entre acidente botrópico e laquético. O tratamento com soro heterólogo específico deve ser realizado precocemente, por via endovenosa. É também importante manter os pacientes hidratados e, no caso de picados por Micrurus, prestar assistência ventilatória adequada

Effect of envenomation on enzymes of brain of albino rats.

Effects of high doses of cobra venom, (150 micrograms/120 +/- 20 g body wt) and viper venom (300 micrograms/120 +/- 20 g body wt) on aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), acetylcholinesterase (ACh) and alkaline phosphatase (ALP) of brain of albino rats were studied. While AST, LDH, ACh and ALP activities increased in both viper and cobra venom treated rats, ALT decreased in both groups compared to control.

Informe especial: estudios tendientes a determinar las posibles propiedades antitumorales del veneno de cobra y del complejo crotoxina A y B / Special report: studies to determine the possible antitumoral properities of Cobra venom and of crotoxin complex A and B

La presente investigación fue encomendada por el CONICET (Consejo Nacional de Investigaciones Cientificas y Técnicas) para determinar las posibles propriedades antineoplásicas del veneno de Cobra (Naja Naja Siamensis) VC) y del Complejo Crotoxina a y B (CCAB), purificado a partir del veneno de Crotalus durissus terificus (cascavel sudamericana). La coordinación de las investigaciones estuvo a cargo de los Dres. Baldi y Mordoh, y la ejecución de la parte experimental fue llevada a cabo por los restantes autores. Se utilizaron diversos sistemas experimentales: 1) lúneas celulares in vitro: de origen murino, normales (3T3 A31), o transformadas (M-A31, Ki-A31 y BP-A31) o de origen humano (adenocarcinoma mamario: MCF-7 y T47D). el efecto de las drogas fue determinado a 1, 10 y 100 ng/ml de VC y CCAB, solas o en combinación. En ninguna de las líneas celulares mencionadas se observaron cambios significativos en la velocidad de crecimiento o en la morfología celular; 2) desarrollo in vivo del Sarcoma 180 (S180) en ratones Balb/c: el VC a dosis de 21 ó 26 ng/g (inyecciones i.p. a los días 10, 11, 20, 21, 30 y 31 post-inoculo tumoral) no afectaron el crecimiento ni la sobrevida de los animales. Dosis de CCAB de 6 ng/g o 9 ng/g ]con el mismo esquema de inoculación anterior) no afectaron el crecimiento de s180 hasta los 25 días de desarrollo tumoral. Entre los días 25 y 30 la dosis de 9 ng/g determinó una evolución tumoral más rápida. La combinación de 13 ng/g VC y 4,5 ng/g CCAB con iguales tiempos de inyección no afectaron el desarrollo tumoral. La histología de los tumores de los animales tratados y controles no evidenció cambios significativos; 3) desarrollo in vivo del carcinoma de Ehrlich en ratones Swiss: se probó el efecto de CCAB (9 ng/g inyectados i.p. a los días 8, 9, 16, y 17) sin obtenerse modificación del desarrollo tumoral;...

Coral snake venoms: mode of action and pathophysiology of experimental envenomation

As cobras corais säo representantes da familia Elapidae nas Américas. Classificam-se em dois gêneros Micruroides e Micrurus. O gênero Micrurus compreende a quase totalidade das espécies de cobra coral e todas as que causam acidentes no homem. Podem-se fazer as seguintes generalizaçöes quanto aos efeitos produzidos por suas peçonhas e a algumas propriedades destas. As peçonhas das cobras corais säo neurotóxicas, causando perda da força muscular e morte por paralisia respiratória. Näo provocam edema local e necrose assim como näo produzem coagulaçäo sanguínea ou hemorragias. A atividade proteolítica das peçonhas de cobras corais é pequena ou nula. Exercem atividade fosfolipase A**2. Näo induzem efeitos nefrotóxicos. Os componentes tóxicos da peçonha das Elapidae säo as neurotoxinas pré-sinápticas, as neurotoxinas pós-sinápticas, as cardiotoxinas e fosfolipases A**2 com atividade mionecrótica ou semelhante à das cardiotoxinas. O modo de açäo das peçonhas de Micrurus frontalis, M. lemniscatus, M. corallinus e M. fulvius foi investigado em preparaçöes neuromusculares isoladas e é aqui exposto. Mostra-se que enquanto as peçonhas de M. frontalis e M. lemniscatus devem conter apenas toxinas que atuam através de ligaçäo com os receptores colinérgicos da placa terminal (neurotoxinas pós-sinápticas), a de M. corallinus atua também na junçäo neurovascular inibindo a liberaçäo de acetilcolina pelos impulsos nervosos e a de M. fulvius induz despolarizaçäo da membrana das fibras musculares. Relatam-se também os efeitos produzidos pelas peçonhas de M. corallinus e M. fulvius in vivo em cäes e os provocados pela peçonha de M. frontalis em cäes e macacos

Mecanismo de acción de miotoxinas aisladas de venenos de serpientes / Mechanism of action of myotoxins isolated from snake venoms

Desde los puntos de vista bioquímico y farmacológico, las miotoxinas aisladas de venenos de serpientes se ubican en cuatro grupos: (1) Fosfolipasas A miotóxicas, (2) miotoxinas básicas de bajo peso molecular, (3) cardiotoxinas de venenos elapídeos y (4) miotoxinas hemorrágicas. Las fosfolipasas miotóxicas notexinam taipoxina, crotoxina y miotoxina de Bothrops asper afectan inicialmente la integridad de la membrana plasmática, induciéndose un influjo de calcio que culmina con la muerte celular. Las miotoxinas básicas de bajo peso molecular crotamina y miotoxina a actúan específicamente en los canales de sodio del sarcolema, induciendo un influjo de sodio que trae como consecuencia despolarización y contracción muscular y vacuolización del retículo sarcoplásmico. Las cardiotoxinas son polipéptidos básicos capaces de desorganizar la estructura de las membranas, siendo su acción miotóxica una consecuencia de la alteración drástica que las mismas inducen en el sarcolema del músculo esquelético. Finalmente, dos componentes hemorrágicos (toxina hemorrágica b y viriditoxina) poseen actividad miotóxica, habiendose sugerido que este efecto es una consecuencia de la isquemia tisular resultante de la acción hemorrágica de estos componentes