Genome-wide Analyses Identify KIF5A as a Novel ALS Gene

To identify novel genes associated with ALS, we undertook two lines of investigation. We carried out a genome-wide association study comparing 20,806 ALS cases and 59,804 controls. Independently, we performed a rare variant burden analysis comparing 1,138 index familial ALS cases and 19,494 controls. Through both approaches, we identified kinesin family member 5A (KIF5A) as a novel gene associated with ALS. Interestingly, mutations predominantly in the N-terminal motor domain of KIF5A are causative for two neurodegenerative diseases: hereditary spastic paraplegia (SPG10) and Charcot-Marie-Tooth type 2 (CMT2). In contrast, ALS-associated mutations are primarily located at the C-terminal cargo-binding tail domain and patients harboring loss-of-function mutations displayed an extended survival relative to typical ALS cases. Taken together, these results broaden the phenotype spectrum resulting from mutations in KIF5A and strengthen the role of cytoskeletal defects in the pathogenesis of ALS.Peer reviewe

A new acidic myotoxic, anti-platelet and prostaglandin I(2) inductor phospholipase A(2) isolated from Bothrops moojeni snake venom

Phospholipase A(2) (PLA(2), EC 3.1.1.4), a major component of snake venoms, specifically catalyzes the hydrolysis of fatty acid ester bonds at position 2 of 1,2-diacyl-sn-3-phosphoglycerides in the presence of calcium. This article reports the purification and biochemical/functional characterization of BmooTX-I, a new myotoxic acidic phospholipase A(2) from Bothrops moojeni snake venom. The purification of the enzyme was carried out through three chromatographic steps (ion-exchange on DEAE-Sepharose, molecular exclusion on Sephadex G-75 and hydrophobic chromatography on Phenyl-Sepharose). BmooTX-I was found to be a single-chain protein of 15,000 Da and pI 4.2. The N-terminal sequence revealed a high homology with other acidic Asp49 PLA(2)S from Bothrops snake venoms. It displayed a high phospholipase activity and platelet aggregation inhibition induced by collagen or ADP. Edema and myotoxicity in vivo were also induced by BmooTX-I. Analysis of myotoxic activity was carried out by optical and ultrastructural microscopy, demonstrating high levels of leukocytary infiltrate. Previous treatment of BmooTX-1 with BPB reduced its enzymatic and myotoxic activities, as well as the effect on platelet aggregation. Acidic myotoxic PLA(2)S from Bothrops snake venoms have been little explored and the knowledge of its structural and functional features will be able to contribute for a better understanding of their action mechanism regarding enzymatic and toxic activities. (C) 2008 Elsevier Ltd. All rights reserved.Fundacao de AmparoPesquisa do Estado de Sao Paulo (FAPESP)Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)Pro-Reitoria de Pesquisa e Pos-Graduaco da Universidade Federal Fluminense (UFF/PROPP), Brazi

Heterologous expression and biochemical and functional characterization of a recombinant alpha-type myotoxin inhibitor from Bothrops alternatus snake

Venomous and non-venomous snakes possess phospholipase A(2) (PLA(2)) inhibitory proteins (PLIs) in their blood serum. This study shows the expression and biochemical and functional characterization of a recombinant alpha inhibitor from Bothrops alternatus snake, named rBaltMIP. Its expression was performed in Pichia pastoris heterologous system, resulting in an active recombinant protein. The expressed inhibitor was tested regarding its ability to inhibit the phospholipase activity of different PLA(2)s, showing slight inhibitions especially at the molar ratios of 1:1 and 1:3 (PLA(2):PLI). rBaltMIP was also effective in decreasing the myotoxic activity of the tested toxins at molar ratios greater than 1:0.4 (myotoxin:PLI). The inhibition of the myotoxic activity of different Asp49 (BthTX-II and PrTX-III) and Lys49 (BthTX-I and PrTX-I) myotoxins was also performed without the prior incubation of myotoxins/inhibitor in order to analyze the real possibility of using snake plasma inhibitors or recombinant inhibitors as therapeutic agents for treating envenomations. As a result, rBaltMIP was able to significantly inhibit the myotoxicity of Lys49 myotoxins. Histopathological analysis of the gastrocnemius muscles of mice showed that the myotoxins are able to induce severe damage to the muscle fibers of experimental animals by recruiting a large number of leukocyte infiltrates, besides forming an intense accumulation of intercellular fluid, leading to local edema. When those myotoxins were incubated with rBaltMIP, a reduction of the damage site could be observed. Furthermore, the cytotoxic activity of Asp49 PLA(2)s and Lys49 PLA(2)-like enzymes on C2C12 cell lines was decreased, as shown by the higher cell viabilities after preincubation with rBaltMIP. Heterologous expression would enable large-scale obtainment of rBaltMIP, thus allowing further investigations for the elucidation of possible mechanisms of inhibition of snake PLA(2)s, which have not yet been fully clarified. (C) 2014 Elsevier Masson SAS. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Purification and Characterization of BmooAi: A New Toxin from Bothrops moojeni Snake Venom That Inhibits Platelet Aggregation

Submitted by EMERSON LEAL ([email protected]) on 2016-04-06T13:14:42Z No. of bitstreams: 1 Purification and Characterization of BmooAi.pdf: 1548419 bytes, checksum: d761c248da6e625f6426c3b0b70fa3ea (MD5)Approved for entry into archive by EMERSON LEAL ([email protected]) on 2016-04-19T13:40:24Z (GMT) No. of bitstreams: 1 Purification and Characterization of BmooAi.pdf: 1548419 bytes, checksum: d761c248da6e625f6426c3b0b70fa3ea (MD5)Made available in DSpace on 2016-04-19T13:40:24Z (GMT). No. of bitstreams: 1 Purification and Characterization of BmooAi.pdf: 1548419 bytes, checksum: d761c248da6e625f6426c3b0b70fa3ea (MD5) Previous issue date: 2014Universidade Federal de Uberlândia. Instituto de Genética e Bioquímica. Uberlândia, MG, Brazil / Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica. Belo Horizonte, MG, Brazil.Universidade Federal de Uberlândia. Instituto de Genética e Bioquímica. Uberlândia, MG, Brazil / Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica. Belo Horizonte, MG, Brazil.Universidade Federal de Uberlândia. Instituto de Genética e Bioquímica. Uberlândia, MG, Brazil / Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica. Belo Horizonte, MG, Brazil.Universidade Federal de Uberlândia. Instituto de Genética e Bioquímica. Uberlândia, MG, Brazil / Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica. Belo Horizonte, MG, Brazil.Universidade Federal de Uberlândia. Instituto de Genética e Bioquímica. Uberlândia, MG, Brazil./ Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica. Belo Horizonte, MG, Brazil.Universidade Federal de Uberlândia. Instituto de Ciências Biomédicas. Uberlândia, MG, Brazil.Universidade Federal de Uberlândia. Instituto de Genética e Bioquímica. Uberlândia, MG, Brazil.Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica. Belo Horizonte, MG, Brazil / Universidade Federal de Uberlândia. Instituto de Ciências Biomédicas. Uberlândia, MG, Brazil.Fundação Oswaldo Cruz. Centro de Estudos de Biomoléculas Aplicadas à Saúde e Núcleo de Saúde. Porto Velho, RO, Brazil / Universidade Federal de Rondônia. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Centro de Estudos de Biomoléculas Aplicadas à Saúde e Núcleo de Saúde. Porto Velho, RO, Brazil / Universidade Federal de Rondônia. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Centro de Estudos de Biomoléculas Aplicadas à Saúde e Núcleo de Saúde. Porto Velho, RO, Brazil / Universidade Federal de Rondônia. Porto Velho, RO, Brazil.Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica. Belo Horizonte, MG, Brazil / Universidade Federal de Uberlândia. Instituto de Ciências Biomédicas. Uberlândia, MG, Brazil.In this paper, we describe the purification/characterization of BmooAi, a new toxin from Bothrops moojeni that inhibits platelet aggregation. The purification of BmooAi was carried out through three chromatographic steps (ion-exchange on a DEAE-Sephacel column, molecular exclusion on a Sephadex G-75 column, and reverse-phase HPLC chromatography on a C2/C18 column). BmooAi was homogeneous by SDS-PAGE and shown to be a single-chain protein of 15,000 Da. BmooAi was analysed by MALDI-TOF Spectrometry and revealed two major components with molecular masses 7824.4 and 7409.2 as well as a trace of protein with a molecular mass of 15,237.4 Da. Sequencing of BmooAi by Edman degradation showed two amino acid sequences: IRDFDPLTNAPENTA and ETEEGAEEGTQ, which revealed no homology to any known toxin from snake venom. BmooAi showed a rather specific inhibitory effect on platelet aggregation induced by collagen, adenosine diphosphate, or epinephrine in human platelet-rich plasma in a dose-dependent manner, whereas it had little or no effect on platelet aggregation induced by ristocetin. The effect on platelet aggregation induced by BmooAi remained active even when heated to 100∘C. BmooAi could be of medical interest as a new tool for the development of novel therapeutic agents for the prevention and treatment of thrombotic disorders

Growing knowledge: an overview of Seed Plant diversity in Brazil

Abstract An updated inventory of Brazilian seed plants is presented and offers important insights into the country's biodiversity. This work started in 2010, with the publication of the Plants and Fungi Catalogue, and has been updated since by more than 430 specialists working online. Brazil is home to 32,086 native Angiosperms and 23 native Gymnosperms, showing an increase of 3% in its species richness in relation to 2010. The Amazon Rainforest is the richest Brazilian biome for Gymnosperms, while the Atlantic Rainforest is the richest one for Angiosperms. There was a considerable increment in the number of species and endemism rates for biomes, except for the Amazon that showed a decrease of 2.5% of recorded endemics. However, well over half of Brazillian seed plant species (57.4%) is endemic to this territory. The proportion of life-forms varies among different biomes: trees are more expressive in the Amazon and Atlantic Rainforest biomes while herbs predominate in the Pampa, and lianas are more expressive in the Amazon, Atlantic Rainforest, and Pantanal. This compilation serves not only to quantify Brazilian biodiversity, but also to highlight areas where there information is lacking and to provide a framework for the challenge faced in conserving Brazil's unique and diverse flora