Immunochemical and biological characterization of monoclonalantibodies against BaP1, a metalloproteinase from Bothrops aspersnake venom

BaP1 is a P-I class of Snake Venom Metalloproteinase (SVMP) relevant in the local tissue damage associated with envenomations by Bothrops asper, a medically-important species in Central America and parts of South America. Six monoclonal antibodies (MoAb) against BaP1 (MABaP1) were produced and characterized regarding their isotype, dissociation constant (Kd), specificity and ability to neutralize BaP1-induced hemorrhagic and proteolytic activity. Two MABaP1 are IgM, three are IgG1 and one is IgG2b. The Kds of IgG MoAbs were in the nM range. All IgG MoAbs recognized conformational epitopes of BaP1 and B. asper venom components but failed to recognize venoms from 27 species of Viperidae, Colubridae and Elapidae families. Clone 7 cross-reacted with three P-I SVMPs tested (moojeni protease, insularinase and neuwiedase). BaP1-induced hemorrhage was totally neutralized by clones 3, 6 and 8 but not by clone 7. Inhibition of BaP1 enzymatic activity on a synthetic substrate by MABaP1 was totally achieved by clones 3 and 6, and partially by clone 8, but not by clone 7. In conclusion, these neutralizing MoAbs against BaP1 may become important tools to understand structure–function relationships of BaP1 and the role of P-I class SVMP in snakebite envenomation.

Preditores espaço-temporais do andar para testes de capacidade funcional em pacientes com doença de Parkinson / Gait spatial and temporal predictors for functional capacity tests in Parkinson's disease patients

OBJETIVOS: Determinar quais parâmetros espaço-temporais são preditores do andar de indivíduos com doença de Parkinson idiopática para os testes de resistência aeróbia e agilidade, propostos pela bateria de testes da American Alliance for Health, Physical Education, Recreation and Dance (AAHPERD). MATERIAIS E MÉTODOS: Foram selecionados seis homens e seis mulheres com comprometimento e estágio da doença em níveis leve e moderado, que realizaram os testes de agilidade e resistência aeróbia, conforme o protocolo da AAHPERD, e andaram 8m sobre uma passarela. Uma câmera digital registrou uma passada central. Marcadores foram fixados no quinto metatarso e na face lateral do calcâneo do membro inferior direito e no primeiro metatarso e na face medial do calcâneo do membro inferior esquerdo. As variáveis dependentes selecionadas foram: tempo nos testes de agilidade e resistência e as variáveis cinemáticas (comprimento da passada - CP, cadência - CAD, duração da passada - DP, duração da fase de duplo suporte - DDS, duração do suporte simples - SS, duração da fase de balanço - DB e velocidade da passada - VP). RESULTADOS: Para agilidade, o teste de Pearson apontou correlação estatisticamente significativa entre as variáveis CP (r=-0,70; p<0,05), CAD (r=-0,72; p<0,01), VP (r=-0,83; p<0,01), DP (r=0,71; p<0,01) e DDS (r=0,90; p<0,01). Para resistência, houve correlação com as variáveis CP (r=-0,67; p<0,05), CAD (r=-0,72; p<0,01), VP (r=-0,82; p<0,01), DP (r=0,71; p<0,01) e DDS (r=0,90; p<0,01). A análise de regressão múltipla revelou que a DDS foi a única variável preditora dos testes de agilidade (R²=0,82; p<0,01) e resistência (R²=0,81; p<0,01). CONCLUSÕES: Observou-se o uso potencial da DDS como parâmetro espaço-temporal do andar preditor do desempenho dos testes de resistência aeróbia e agilidade em pacientes com doença de Parkinson idiopática.

OBJECTIVE: To determine which spatial and temporal parameters are predictors of the gait pattern of individuals with idiopathic Parkinson's disease, for the aerobic endurance and agility tests included in the battery of tests of the AAHPERD protocol. METHODS: Six men and six women with mild and moderate impairment and disease stage were selected. They performed agility and aerobic endurance test in accordance with the AAHPERD protocol, and walked 8 m on a walkway. A digital video camera recorded one central stride. Markers were attached to the fifth metatarsal and lateral face of the calcaneus of the right leg and to the first metatarsal and medial face of the calcaneus of the left leg. The dependent variables selected were the time taken in the agility and endurance tests and the kinematic variables: stride length (SL), cadence (CAD), stride time (ST), double support time (DS), single support time (SS), swing time (SW) and stride velocity (SV). RESULTS: For agility, Pearson's test showed statistically significant correlations with SL (r=-0.70; p<0.05), CAD (r=-0.72; p<0.01), SV (r=-0.83; p<0.01), ST (r=0.71; p<0.01) and DS (r=0.90; p<0.01). For endurance, there were correlations with SL (r=-0.67; p<0.05), CAD (r=-0.72; p<0.01), SV (r=-0.82; p<0.01), ST (r=0.71; p<0.01), and DS (r=0.90; p<0.01). Multiple regression analysis revealed that DS was the only variable that predicted performance in both the agility (R²=0.82; p<0.01) and the endurance (R²=0.81; p<0.01) tests. CONCLUSIONS: It was observed that DS measure has a potential use as a kinematic parameter of gait that predicts the performance in agility and aerobic endurance tests in patients with idiopathic Parkinson's disease.