Insights of local tissue damage and regeneration induced by BnSP-7, a myotoxin isolated from Bothrops (neuwiedi) pauloensis snake venom.

Envenomations caused by Bothrops snake venoms are characterized by prominent local tissue damage due to myonecrosis, hemorrhage, edema and acute muscle damage which is widely correlated with phospholipases A2 (PLA2). In the present study, the progression of local tissue damage and inflammation induced by BnSP-7, a myotoxin isolated from Bothrops (neuwiedi) pauloensis snake venom, was evaluated. Local tissue damages characterized by edema, necrosis and inflammation were evaluated until 24 h after inoculation of BnSP-7. The regeneration of myofibers, analyzed by light microscopy, was observed from 72 h to 2 weeks post-inoculation of toxin. MMP-2 was expressed in gastrocnemius muscle at all time points tested, while the expression of MMP-9 increased expressively at the same time interval of regenerating muscle, suggesting the involvement of MMP-9 in the regeneration process. The production of pro-inflammatory cytokines was also increased, whereas IL-1 beta showed the highest level. Modification of BnSP-7 with BPB decreased the release of IL-8, IL-6 and IL-1 beta when compared to native BnSP-7. These data suggest that BnSP-7 acts as pro-inflammatory incentives (mediators), inducing MMP and cytokine production from the inflammatory and satellite cells, and thus it may play an important role in inflammatory process and, consequently, in the evolution of local tissue damage and regeneration.

Characterization of inflammatory reaction induced by neuwiedase, a P-I metalloproteinase isolated from Bothrops neuwiedi venom.

The Snake Venom Metalloproteinases (SVMPs) play a relevant role in the multifactorial inflammatory response induced by Bothrops envenomations. Neuwiedase, an SVMP isolated from Bothrops neuwiedi venom, is devoid of hemorrhagic activity on skin tests, but is able to induce myonecrosis and degrade fibrinogen, fibrin, type I collagen, fibronectin and laminin. In this study, we analyzed the inflammatory reaction induced by neuwiedase in gastrocnemius muscle, with special focus on cytokines release. Our results showed clear evidence of inflammatory infiltrate in the gastrocnemius muscle and an increase of MMP-9, and the cytokines KC, IL-1 beta and IL-6 in the early periods after toxin injection. The cytokine release was also evaluated in inflammatory and muscular cell culture. Both murine peritoneal adherent cells (MPACs) and muscle cells (C2C12) released pro-inflammatory cytokines after stimulus with neuwiedase. MPACs showed increased production of KC, IL-1 beta and IL-6 in the cell culture supernatant while in C2C12, the predominant chemokine expressed was KC. These data reinforce the importance of SVMPs in the inflammatory response caused by envenomation and point out the role of muscle cells in this event by releasing pro-inflammatory mediators able to attract leukocytes to the muscle, thus starting and amplifying the setting of the inflammatory reaction.

Análise microbiológica de queijos tipo minas frescal, comercializados em duas feiras-livres na cidade de Uberlândia, MG / Microbiological analysis of fresh cheese type mines, commercialized in two free fairs in the city of Uberlandia, Brazil

Com o objetivo de avaliar as condições higiênico-sanitárias do queijo Minas Frescal produzido artesanalmente em Uberlândia – MG, foi realizada análise microbiológica de dez amostras obtidas em duas feiras-livres nas quais são comercializados. As amostras foram analisadas em laboratório, através da técnica do NMP, para coliformes fecais. Oito das dez amostras (A, E, D, F, G, H, I e J) apresentaram-se em conformidade com os padrões exigidos pela legislação (RDC n° 12/ANVISA). Segundo o padrão de amostragens e métodos adotados, as amostras apresentaram condições higiênico-sanitárias satisfatórias, de acordo com o padrão legal vigente, indicando baixo nível de contaminação direta ou indiretamente com material fecal, sendo classificadas como produtos próprios para o consumo. Entretanto, os resultados das análises das amostras (B e C) não foram exatos, impossibilitando uma conclusão definitiva sobre o índice de coliformes fecais presente nestas amostras de queijos.