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1.
Insect Mol Biol ; 26(1): 25-34, 2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-27743460

RESUMO

Loxosceles intermedia venom comprises a complex mixture of proteins, glycoproteins and low molecular mass peptides that act synergistically to immobilize envenomed prey. Analysis of a venom-gland transcriptome from L. intermedia revealed that knottins, also known as inhibitor cystine knot peptides, are the most abundant class of toxins expressed in this species. Knottin peptides contain a particular arrangement of intramolecular disulphide bonds, and these peptides typically act upon ion channels or receptors in the insect nervous system, triggering paralysis or other lethal effects. Herein, we focused on a knottin peptide with 53 amino acid residues from L. intermedia venom. The recombinant peptide, named U2 -sicaritoxin-Li1b (Li1b), was obtained by expression in the periplasm of Escherichia coli. The recombinant peptide induced irreversible flaccid paralysis in sheep blowflies. We screened for knottin-encoding sequences in total RNA extracts from two other Loxosceles species, Loxosceles gaucho and Loxosceles laeta, which revealed that knottin peptides constitute a conserved family of toxins in the Loxosceles genus. The insecticidal activity of U2 -SCTX-Li1b, together with the large number of knottin peptides encoded in Loxosceles venom glands, suggests that studies of these venoms might facilitate future biotechnological applications of these toxins.


Assuntos
Aranha Marrom Reclusa/genética , Miniproteínas Nó de Cistina/química , Inseticidas/análise , Diester Fosfórico Hidrolases/química , Venenos de Aranha/química , Sequência de Aminoácidos , Animais , Sequência de Bases , Aranha Marrom Reclusa/metabolismo , Sequência Conservada , Miniproteínas Nó de Cistina/biossíntese , Miniproteínas Nó de Cistina/genética , Miniproteínas Nó de Cistina/isolamento & purificação , Dípteros , Eletroforese em Gel de Poliacrilamida , Escherichia coli , Dados de Sequência Molecular , Proteoma , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Testes de Toxicidade , Transcriptoma
2.
Lipids ; 48(9): 889-97, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23743575

RESUMO

Episodes of acute exacerbation are the major clinical feature of asthma and therefore represent an important focus for developing novel therapies for this disease. There are many reports that the n-3 fatty acids found in fish oil exert anti-inflammatory effects, but there are few studies of the action of fish oil on airway smooth muscle (ASM) function. In the present investigation, we evaluated the effect of fish oil supplementation on smooth muscle force of contraction in ovalbumin-induced asthmatic Wistar rats, and its consequences on static lung compliance, mucus production, leukocyte chemotaxis and production of proinflammatory cytokines. Fish oil supplementation suppressed the infiltration of inflammatory cells into the lung in asthmatic animals (2.04 ± 0.19 × 10(6) cells vs. 3.33 ± 0.43 × 10(6) cells in the control asthmatic group; P < 0.05). Static lung compliance increased with fish oil supplementation in asthmatic rats (0.640 ± 0.053 mL/cm H2O vs. 0.399 ± 0.043 mL/cm H2O; P < 0.05). However, fish oil did not prevent asthma-associated lung eosinophilia and did not affect the concentrations of tumor necrosis factor-α and interleukin-1ß in lung tissue or the proportion of the airways obliterated with mucus. Fish oil had no effect on the force of contraction in asthmatic rats in response to acetylcholine (3.026 ± 0.274 mN vs. 2.813 ± 0.364 mN in the control asthmatic group). In conclusion, although fish oil exerts some benefits in this model of asthma, its effectiveness appears to be limited by an inefficient action on airway smooth muscle function.


Assuntos
Asma/fisiopatologia , Óleos de Peixe/farmacologia , Músculo Liso/efeitos dos fármacos , Traqueia/efeitos dos fármacos , Acetilcolina/farmacologia , Análise de Variância , Animais , Asma/induzido quimicamente , Líquido da Lavagem Broncoalveolar/citologia , Broncodilatadores/farmacologia , Suplementos Nutricionais , Eosinófilos/efeitos dos fármacos , Eosinófilos/patologia , Óleos de Peixe/administração & dosagem , Técnicas In Vitro , Interleucina-1beta/metabolismo , Isoproterenol/farmacologia , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Pulmão/patologia , Complacência Pulmonar/efeitos dos fármacos , Linfócitos/efeitos dos fármacos , Linfócitos/patologia , Macrófagos/efeitos dos fármacos , Macrófagos/patologia , Masculino , Músculo Liso/fisiopatologia , Neutrófilos/efeitos dos fármacos , Neutrófilos/patologia , Ovalbumina , Ratos , Ratos Wistar , Traqueia/fisiopatologia , Fator de Necrose Tumoral alfa/metabolismo , Vasodilatadores/farmacologia
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