Isolation and characterization of four type 2 ribosome inactivating pulchellin isoforms from Abrus pulchellus seeds

Abrus pulchellus seeds contain at least seven closely related and highly toxic type 2 ribosome-inactivating pulchellins, each consisting of a toxic A-chain linked to a sugar binding B-chain. In the present study, four pulchellin isoforms (termed P I, P II, P III and P IV) were isolated by affinity, ion exchange and chromatofocusing chromatographies, and investigated with respect to toxicity and sugar binding specificity. Half maximal inhibitory concentration and median lethal dose values indicate that P I and P II have similar toxicities and that both are more toxic to cultured HeLa cells and mice than P III and P IV. Interestingly, the secondary structural characteristics and sugar binding properties of the respective pairs of isoforms correlate well with the two toxicity levels, in that P I/P II and P III/P IV form two specific subgroups. From the deduced amino acids sequences of the four isoforms, it is clear that the highest similarity within each subgroup is found to occur within domain 2 of the B-chains, suggesting that the disparity in toxicity levels might be attributed to subtle differences in B-chain-mediated cell surface interactions that precede and determine toxin uptake pathways.

Atividade antitumoral e imunológica de abrina e pulchelina em tumor mamário murino

O termo “câncer” corresponde ao conjunto de cerca de 100 doenças que têm em comum o crescimento desordenado de células que invadem os tecidos e órgãos, podendo metastatisar para outras regiões do corpo. Os tumores de mama e colo do útero são os mais frequentes no sexo feminino. Neste estudo, avaliou-se o efeito tóxico de injeções intratumorais de abrina e pulchelina com ou sem β-D-galactose sobre o desenvolvimento do tumor mamário murino, verificando sua influência sobre o sistema imune. Proteínas inativadoras de ribossomos (RIPs) abrina, obtida de sementes de Abrus precatorius, e pulchelina, de sementes maduras de Abrus pulchellus subsp. tenuiflorus, foram utilizadas e como droga controle foi usada a Doxorrubicina. As RIPs foram administradas em camundongos fêmeas Balb/c. A partir dos tumores retirados dos animais em estudo, verificou-se o percentual de inibição do crescimento tumoral, medindo-se o tamanho e os pesos dos tumore. A partir de culturas de macrófagos obtidos dos animais de estudo, avaliou-se a produção de NO, TNF-α e IL-12 pelas RIPs na presença ou ausência de β-D-galactose. A IL-10 foi quantificada a partir de linfócitos esplênicos. A viabilidade celular foi verificada quando as células foram sujeitas às ações das RIPs. Com base nos resultados obtidos, observou-se que as RIPs não apresentaram potencial antitumoral, pois não houve redução do tamanho do tumor em relação ao controle, exceto pela abrina (p<0,05). Contudo, verificou-se que houve um possível efeito inibitório da toxicidade de abrina e pulchelina pela galactose sobre as células tumorais. As substâncias testadas (abrina, pulchelina e doxorrubicina) nas concentrações utilizadas nos testes de citotoxicidade ...

The role of the C-terminal region of pulchellin A-chain in the interaction with membrane model systems

Pulchellin is a Ribosome Inactivating Protein containing an A-chain (PAC), whose toxic activity requires crossing the endoplasmic reticulum (ER) membrane. In this paper, we investigate the interaction between recombinant PAC (rPAC) and Langmuir monolayers of dipalmitoyl phosphatidyl glycerol (DPPG), which served as membrane model. Three catalytically active, truncated PACs with increasing deletion of the C-terminal region, possessing 244,239 and 236 residues (rPAC(244), rPAC(239) and rPAC(236)), were studied. rPAC had the strongest interaction with the DPPG monolayer, inducing a large expansion in its surface pressure-area isotherm. The affinity to DPPG decreased with increased deletion of the C-terminal region. When the C-terminal region was deleted completely (rPAC(236)), the interaction was recovered, probably because other hydrophobic regions were exposed to the membrane. Using Polarization Modulated-Infrared Reflection Absorption Spectroscopy (PM-IRRAS) we observed that at a bare air/water interface rPAC comprised mainly alpha-helix structures, the C-terminal region had unordered structures when interacting with DPPG. For rPAC(236) the alpha-helices were preserved even in the presence of DPPG. These results confirm the importance of the C-terminal region for PAC-ER membrane interaction. The partial unfolding only with preserved C-terminal appears a key step for the protein to reach the cytosol and develop its toxic activity. (C) 2011 Elsevier B.V. All rights reserved.

IMUNIDADE CRUZADA PELAS SEMENTES DE Abrus precatorius E Ricinus communis EM BOVINOS

Cinco bovinos imunizados contra a ação tóxica das sementes de Abrus precatorius L. ("tento", "jiquiriti") não adoeceram ou somente levemente pela administração das sementes de Ricinus communis L. ("mamona"), em doses que em bovinos que antes nunca ingeriram sementes de A. precatorius ou R. communis, causaram intoxicação de intensidade de grau moderado a acentuado ou até a morte. Um sexto bovino, que não ficou bem imunizado contra a ação tóxica das sementes de A. precatorius, adoeceu em grau acentuado pela administração de dose elevada das sementes de R. communis. Já dos cinco bovinos imunizados contra a ação tóxica das sementes de R. communis quatro adoeceram em grau acentuado, oquinto em grau moderado, pela administração das sementes de A. precatorias em doses que em bovinos que antes nunca ingeriram sementes de R. communis ou A. precatorius causaram intoxicação de intensidade leve a acentuada. Estes resultados permitem concluir que bovinos imunizados contra a ação tóxica das sementes de A. precatorius são resistentes à ação tóxica das sementes de R. communis, mas que o contrário não ocorre, isto é, bovinos imunizados contra a ação tóxica das sementes de R. communis, não se mostraram protegidos contra a intoxicação por A. precatorius. Estudos anteriores por outros autores mostraram que as toxalbuminas de A. precatorius e R. communis, respectivamente abrina e ricina, são diferentes do ponto de vista antigênico. Uma explicação para a divergência desses resultados com os nossos poderia estar no fato de que no presente estudo foram usados poligástricos que receberam as sementes por via oral, enquanto que nos estudos anteriores foram usados monogástricos em que as sementes ou as toxinas foram aplicadas por via parenteral. A administração de folhas frescas ou do pericarpo do fruto de R. communis a bovinos imunizados contra a ação das sementes desta planta tiveram o mesmo efeito tóxico que em animais não imunizados, demonstrando que a imunidade conferida pela ricina não inibe a ação da ricinina, o principio tóxico das folhas e do pericarpo.

Redescription of Hypsiboas semiguttatus, with the description of a new species of the Hypsiboas pulchellus group

We redescribe Hypsiboas semiguttatus (Lutz, 1925) and describe its tadpole and vocalization. We also describe a new species, previously confused with H. semiguttatus from Misiones, Argentina, and potentially from Brazil. This new species differs from H. semiguttatus in being more robust, having a tadpole with complete marginal papulation (no anterior gap), and a labial tooth row formula of 3/5. We discuss the status of populations previously assigned to H. semiguttatus and discuss the relationships of the new species within the Hypsiboas pulchellus group. © 2007 by the American Society of Ichthyologists and Herpetologists.

Vocalizations and comments on the relationships of Hypsiboas ericae (Amphibia, Hylidae)

The vocalizations of Hypsiboas ericae (Caramaschi & Cruz, 2000) are described and new information on the external morphology and osteology of the species are presented. H. ericae presents a bony spine in the prepolex and the individuals can present green or brown dorsal color, as other species of the Hypsiboas pulchellus (Duméril & Bibron, 1841) species group. The vocalizations of H. ericae are similar to the vocalizations of Hypsiboas bischoffi (Boulenger, 1887), Hypsiboas guentheri (Boulenger, 1886), and other species in the H. polytaenius (Cope, 1870 "1869") clade of the H. pulchellus species group, but some osteological aspects are different to those found in the majority of the species of this group.

Dormancy as exaptation to protect mimetic seeds against deterioration before dispersal

Mimetic seeds simulate the appearance of fleshy fruits and arilled seeds without producing nutritive tissues as a reward for seed dispersers. In this strategy of seed dispersal, seeds may remain attached to the mother plant for long periods after maturity, increasing their availability to naive seed dispersers. The hypothesis that seed coat impermeability in many tropical Fabaceae with mimetic seeds serves as an exaptation to protect the seeds from deterioration and rotting while awaiting dispersal was investigated. Seed coat impermeability was evaluated in five mimetic-seeded species of tropical Fabaceae in south-eastern Brazil (Abarema langsdorffii, Abrus precatorius, Adenanthera pavonina, Erythrina velutina and Ormosia arborea) and in Erythrina speciosa, a `basal` species in its genus, which has monochromatic brown seeds and no mimetic displays. Seed hardness was evaluated as a defence against accelerated ageing (humid chamber at 41 degrees C for 144 h). Seed development and physiological potential of O. arborea was evaluated and the effect of holding mature seeds in pods on the mother plant in the field for a period of 1 year under humid tropical conditions was compared with seeds stored under controlled conditions (15 degrees C and 40 % relative air humidity). All five mimetic-seeded species, and E. speciosa, showed strong coat impermeability, which protected the seeds against deterioration in accelerated ageing. Most O. arborea seeds only became dormant 2 months after pod dehiscence. Germination of seeds after 1 year on the plant in a humid tropical climate was 56 %, compared with 80 % for seeds stored in controlled conditions (15 degrees C, 45 % relative humidity). Seedling shoot length after 1 year did not differ between seed sources. Dormancy acts in mimetic-seeded species as an exaptation to reduce seed deterioration, allowing an increase in their effective dispersal period and mitigating the losses incurred by low removal rates by naive avian frugivores.

Vocalizations and comments on the relationships of Hypsiboas ericae (Amphibia, Hylidae)

The vocalizations of Hypsiboas ericae (Caramaschi & Cruz, 2000) are described and new information on the external morphology and osteology of the species are presented. H. ericae presents a bony spine in the prepolex and the individuals can present green or brown dorsal color, as other species of the Hypsiboas pulchellus (Duméril & Bibron, 1841) species group. The vocalizations of H. ericae are similar to the vocalizations of Hypsiboas bischoffi (Boulenger, 1887), Hypsiboas guentheri (Boulenger, 1886), and other species in the H. polytaenius (Cope, 1870 "1869") clade of the H. pulchellus species group, but some osteological aspects are different to those found in the majority of the species of this group.

Sinonímias e descrições em Adetus LeConte (Coleoptera, Cerambycidae, Lamiinae, Apomecynini)

Based on the photographs (color slides) taken from the type specimens of Adetus species, the following synonyms are established: Adetus subcostatus Aurivillius, 1900 = A. auratoides Breuning, 1943 syn. nov.; A. leucostigma Bates, 1880 = A. venezuelensis Breuning, 1942 syn. nov.; A. pulchellus (Thomson, 1868) = A. laterialbus Breuning, 1942 syn. nov.; A. irregularis (Breuning,1939) = A. gracilis Breuning, 1940 syn. nov.; A. flavescens Melzer, 1934 = A. strandi Breuning, 1940 syn. nov. = A. mediofasciculatus Breuning, 1943 syn. nov.; A. modestus Melzer, 1934 = Atimuropsis densepunctata Breuning, 1939 syn. nov. New species described: Adetus multifasciatus sp. nov. (Brazil, São Paulo); A. lineatus sp. nov. (Brazil, Distrito Federal); A. linsleyi sp. nov. (Ecuador, Los Ríos); A. pacaruaia sp. nov. (Peru, Huanuco), and A. pinima sp. nov. (Peru, Junin). Adetus marmoratus Breuning, 1942 is redescribed.

Cramoll 1,4 lectin increases ROS production, calcium levels, and cytokine expression in treated spleen cells of rats

This study reports the in vivo stimulatory effects of Cramoll 1,4 on rat spleen lymphocytes as evidenced by an increase in intracellular reactive oxygen species (ROS) production, Ca(2+) levels, and interleukin (IL)-1 beta expression. Cramoll 1,4 extracted from seeds of the Leguminosae Cratylia mollis Mart., is a lectin with antitumor and lymphocyte mitogenic activities. Animals (Nine-week-old male albino Wistar rats, Rattus norvegicus) were treated with intraperitoneal injection of Cramoll 1,4 (235 mu g ml(-1) single dose) and, 7 days later, spleen lymphocytes were isolated and analyzed for intracellular ROS, cytosolic Ca(2+), and IL-6, IL-10, and IL-1 mRNAs. Cell viability was investigated by annexin V-FITC and 7-amino-actinomycin D staining. The data showed that in lymphocytes activated by Cramoll 1,4 the increase in cytosolic and mitochondrial ROS was related to higher cytosolic Ca(2+) levels. Apoptosis and necrosis were not detected in statistically significant values and thus the lectin effector activities did not induce lymphocyte death. In vivo Cramoll 1,4 treatment led to a significant increase in IL-1 beta but IL-6 and -10 levels did not change. Cramoll 1,4 had modulator activities on spleen lymphocytes and stimulated the Th2 response.

Vocalizations and comments on the relationships of Hypsiboas ericae (Amphibia, Hylidae)

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

A new species of Hypsiboas from the Atlantic forest of southeastern Brazil (Amphibia : Anura : Hylidae)

We describe a new species of Hypsiboas of the Hypsiboas pulchellus species group from highland streams of the Atlantic Forest of southeastern Brazil. Vocalizations, egg-mass, and tadpole are also described. We compared the new species with other species of the Hypsiboas pulchellus species group. The new species is most similar to Hypsiboas semiguttatus, its sister species, and Hypsiboas curupi, from which it differs in advertisement call and larval morphology. We provide information on natural history, phylogenetic relationships, embryos, geographic distribution, and conservation.

Comparative larval morphology of eight species of Hypsiboas Wagler (Amphibia, Anura, Hylidae) from Argentina and Uruguay, with a review of the larvae of this genus

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

A review of antifertility folkloric plants tested in laboratory animals

The use of natural active principals is widespread among a great proportion of the rural population, or by people who do not have easy access to medical assistance. These active principles are used as food or medicines, and even for purposes of contraception. It becomes necessary to establish a relationship between the folklore habits and current information on the nature of anti-fertility substances, and knowledge of their mechanisms. Anti-fertility agents may exert their actions in a number of areas, (hypothalamus, anterior pituitary, oviduct, uterus, and vagina), inhibiting synthesis and/or liberation of hormones (follicle-stimulating, luteinizing, and steroid hormones), ovulation, ovum transportation, and implantation process. Therefore, a review of literature was carried out, including of several plants used by women as abortifacient and anti-fertility agents to compare their effects with those obtained among laboratory animals.