A rapid procedure for the isolation of the Lys-49 myotoxin II from Bothrops moojeni (caissaca) venom: Biochemical characterization, crystallization, myotoxic and edematogenic activity

A, M. Soares, V, M, Rodrigues, M. I. Homsi-Brandeburgo, M. H. Toyama, F, R, Lombardi, K. Arni and J. R, Giglio. A rapid procedure for the isolation of the Lys-49 myotoxin II from Bothrops moojeni (caissaca) venom: Biochemical characterization, crystallization, myotoxic and edematogenic activity. Toxicon 36, 503-514, 1998.-Bothrops moojeni snake venom was fractionated on a CM-Sepharose column which was previously equilibrated with 0.05 M ammonium bicarbonate buffer at pH 8.0 and subsequently eluted with an ammonium bicarbonate concentration gradient from 0.05 to 0.5 M at constant pH (8.0) and temperature (25 degrees C). The fraction which eluted last (M-VI) showed, after direct lyophilization, a single band by polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE, indicating an approximate M,. of 14 000 and 77 000, in the presence and absence of dithiothreitol, respectively. Its amino acid composition revealed a high level of hydrophobic and basic amino acids as well as 13 half-cystine residues. Its isoelectric point and extinction coefficient (E-1.0cm(1.0mg/ml) at 278 nm and pH 7.0) were 8.2 and 1.170, respectively. M-VI was devoid of phospholipase A(2) (PLA(2)) activity on egg yolk, as well as of hemorrhagic, anticoagulant and coagulant activities, but could induce drastic necrosis on skeletal muscle fibres as well as rapid and transient edema on the rat paw. Its N-terminal sequence: SLFELGKMILQETGKNPAKSYGVYGCNCGVGGRGKPKDATDRCCYVHKCCYK.... revealed high homology with other Lys 49 PLA(2)-like myotoxins from other bothropic venoms. Orthorhombic crystals of M-VI? which diffracted to a maximal resolution of 1.6 Angstrom. were obtained and indicated the presence of a dimer in the asymmetrical unit. (C) 1998 Elsevier B.V. Ltd. All rights reserved.

Purification and biochemical characterization of two xylanases produced by Aspergillus caespitosus and their potential for kraft pulp bleaching

Two extracellular xylanases produced by the thermotolerant fungus Aspergillus caespitosus grown in sugar cane bagasse were purified and characterized. Estimated molecular masses were 26.3 and 27 kDa (xyl I); 7.7 and 17.7 kDa (xyl II) for gel filtration and SDS-PAGE, respectively. Optimal temperature for both xylanases was 50-55°C. Optimal pH was 6.5-7.0 for xyl I, and 5.5-6.5 for xyl II. The thermostability (T half) at 55°C was 27.3 min (xyl I) and >90 min (xyl II). Xylanase activity was inhibited by several ions. β-mercaptoethanol activated 59 and 102% xyl I and xyl II activities, respectively. These enzymes preferentially hydrolyzed birchwood xylan, and the K m and V max values were 2.5 mg/ml and 1679 U/mg protein (xyl I), and 3.9 mg/ml and 113 U/mg protein (xyl II). The action of both xylanases mainly that of xyl II, on kraft pulp reduced kappa number and increased pulp viscosity. © 2004 Elsevier Ltd. All rights reserved.

Status and potential of commercial bioprospecting activities in Latin America and the Caribbean

Includes bibliography

Combined effects of age and diet-induced obesity on biochemical parameters and cardiac energy metabolism in rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Biochemical responses in armored catfish (Pterygoplichthys anisitsi) after short-term exposure to diesel oil, pure biodiesel and biodiesel blends

Biodiesel fuel is gradually replacing petroleum-based diesel oil use. Despite the biodiesel being considered friendlier to the environment, little is known about its effects in aquatic organisms. In this work we evaluated whether biodiesel exposure can affect oxidative stress parameters and biotransformation enzymes in armored catfish (Pterygoplichthys anisitsi, Loricariidae), a South American endemic species. Thus, fish were exposed for 2 and 7d to 0.01mLL-1 and 0.1mLL-1 of pure diesel, pure biodiesel (B100) and blends of diesel with 5% (B5) and 20% (B20) biodiesel. Lipid peroxidation (malondialdehyde) levels and the activities of the enzymes glutathione S-transferase, superoxide dismutase, catalase and glutathione peroxidase were measured in liver and gills. Also, DNA damage (8-oxo-7, 8-dihydro-2'-deoxyguanosine) levels in gills and 7-ethoxyresorufin-O-deethylase activity in liver were assessed. Pure diesel, B5 and B20 blends changed most of the enzymes tested and in some cases, B5 and B20 induced a higher enzyme activity than pure diesel. Antioxidant system activation in P. anisitsi was effective to counteract reactive oxygen species effects, since DNA damage and lipid peroxidation levels were maintained at basal levels after all treatments. However, fish gills exposed to B20 and B100 presented increased lipid peroxidation. Despite biodiesel being more biodegradable fuel that emits less greenhouse gases, the increased lipid peroxidation showed that biofuel and its blends also represent hazards to aquatic biota. © 2013 Elsevier Ltd.

Biochemical, functional, structural and phylogenetic studies on Intercro, a new isoform phospholipase A2 from Crotalus durissus terrificus snake venom

Crotoxin is a neurotoxin from Crotalus durissus terrificus venom that shows immunomodulatory, anti-inflammatory, antimicrobial, antitumor and analgesic activities. Structurally, this toxin is a heterodimeric complex composed by a toxic basic PLA2 (Crotoxin B or CB) non-covalently linked to an atoxic non-enzymatic and acidic component (Crotapotin, Crotoxin A or CA). Several CA and CB isoforms have been isolated and characterized, showing that the crotoxin venom fraction is, in fact, a mixture of different molecules derived from the combination of distinct subunit isoforms. Intercro (IC) is a protein from the same snake venom which presents high similarity in primary structure to CB, indicating that it could be an another isoform of this toxin. In this work, we compare IC to the crotoxin complex (CA/CB) and/or CB in order to understand its functional aspects. The experiments with IC revealed that it is a new toxin with different biological activities from CB, keeping its catalytic activity but presenting low myotoxicity and absence of neurotoxic activity. The results also indicated that IC is structurally similar to CB isoforms, but probably it is not able to form a neurotoxic active complex with crotoxin A as observed for CB. Moreover, structural and phylogenetic data suggest that IC is a new toxin with possible toxic effects not related to the typical CB neurotoxin. © 2013.

Genetic and biochemical markers of hydroxyurea therapeutic response in sickle cell anemia

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

The Gastroprotective Effect of Menthol: Involvement of Anti-Apoptotic, Antioxidant and Anti-Inflammatory Activities

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Isolation and biochemical characterization of a gamma-type phospholipase A2 inhibitor from Crotalus durissus collilineatus snake serum

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Biochemical characterisation and application of lipases produced by Aspergillus sp on solid-state fermentation using three substrates

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Rutin ameliorates glycemic index, lipid profile and enzymatic activities in serum, heart and liver tissues of rats fed with a combination of hypercaloric diet and chronic ethanol consumption

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Hematological and biochemical changes in a Phrynops geoffroanus (Schweigger, 1812) population in response to environmental stressors

Turtles are among the most endangered vertebrate groups, and the main threats to populations are environmental pollution and habitat degradation. The species Phrynops geoffroanus, popularly known as “Geoffroy’s side-necked turtle”, has proliferated in polluted environments, where adverse conditions could influence their living habits and physiological condition. Studies that monitor the effects of environmental pollution are key to understanding the species’ biology and designing effective conservation strategies. Thus, the analysis of hematological and biochemical parameters has been shown to be important in assessing the health of wild animals and risks for the animal and ecosystem. This study aimed to assess the environmental influence on the physiology of a P. geoffroanus population through the evaluation of antioxidant status and responses to environmental stressors, compared to specimens from a place under controlled conditions. Blood samples of 60 specimens were collected, 30 from the Felicidade Stream, polluted environment, within the city of São José do Rio Preto, and 30 from the “Reginaldo Uvo Leone” breeding farm, Tabapuã, SP, a place under controlled conditions, whose samples constituted the control group. They were evaluated by hemogram and by determining thiobarbituric acid reactive species (TBARS), Trolox-equivalent antioxidant capacity (TEAC) and the activities of the antioxidant enzymes catalase and glucose-6-phosphate dehydrogenase (G6PDH). There was a wide variation in hematological parameters of P. geoffroanus from the urban environment. The red blood cell count and hemoglobin values were significantly less than those observed in animals from the breeding farm (P = 0.0004; P = 0.0371, respectively). There was a significant increase in the number of thrombocytes (P < 0.0001) and leukocytes (P < 0.0001) in the animals from Felicidade Stream. The stress indices were similar between the two groups (P = 0.4077). TBARS levels showed the cytotoxic potential of compounds in the urban environment, whose animals had elevated levels of lipid peroxidation (P < 0.0001), despite showing a response to environmental damages with increase in antioxidant capacity, as demonstrated by the TEAC assay (P = 0.0207). The lower catalase enzyme activity noted in individuals from the urban environment (P = 0.000184) could be due to the presence of inhibitory compounds. On the other hand, G6PDH activity was higher (P = 0.002962), where this enzyme acts in the generation of NADPH, which is used in several detoxification pathways. We conclude that environmental contamination can increase oxidative damages and generate physiological changes in this species. These data are very useful for the conservation of P. geoffroanus and turtles in general, and confirm that these techniques are effective in monitoring natural regions and that P. geoffroanus can serve as an environmental contamination bioindicator.

Cadmium exposure inhibits MMP2 and MMP9 activities in the prostate and testis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Biochemical responses, morphometric changes, genotoxic effects and CYP1A expression in the armored catfish Pterygoplichthys anisitsi after 15 days of exposure to mineral diesel and biodiesel

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Biochemical responses in mussels Perna perna exposed to diesel B5

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)