EP80317 Restrains Inflammation and Mortality Caused by Scorpion Envenomation in Mice

Over 1 million cases of scorpion stings are estimated every year, whereas current treatment is limited to antivenom serum combined with supportive therapy. Tityus serrulatus scorpion venom (TsV) is composed of diverse molecules, including toxins that induce a catecholamine storm and mediate classical symptoms of scorpion envenomation. However, the same toxins promote an intense inflammatory response coordinated by innate immune cells, such as macrophages, contributing significantly to the lung edema and mortality caused by TsV injection. Macrophages sense TsV via innate immune receptors, including TLR2, TLR4, and CD14 that promote inflammation and mortality via PGE2/cAMP/PKA/NF-κB/IL-1β axis. The scavenger receptor CD36 also recognizes TsV, but in contrast to the other receptors, it drives the production of leukotriene B4 (LTB4). This lipid mediator operates via BLT1 receptor to reduce cAMP production and consequently IL-1β release, which results in resistance to fatal outcomes of experimental scorpion envenomation. EP80317 is an hexapeptide that serves as a ligand for CD36 and features protective effects under conditions such as atherosclerosis and vascular inflammation. In this study, we evaluated the effects of EP80317 treatment during experimental scorpion envenomation. EP80317 treatment suppressed mouse peritoneal macrophage production of IL-1β, IL-6, tumor necrosis factor (TNF-α), CCL3, and PGE2in vitro. EP80317 treatment also boosted the production of LTB4 and IL-10 in response to TsV. Importantly, EP80317 restrained lung inflammation and mortality caused by TsV in vivo. Taken together, these data indicate a strong therapeutic potential of EP80317 as a supportive treatment to control inflammation induced by scorpion envenomation

Vacinas: importância, verdades e mitos – uma revisão bibliográfica

Introduction: Since its implementation, the vaccination process has been questioning and criticizing its effectiveness, immunological functioning and results. From this, even though it is an efficient public policy in the fight against infectious diseases, we are aware of the non-achievement, which harms the prevention of disease morbidity and mortality. Methods: Bibliographical review using as databases the Google academic, Medline, Center for Biotechnology Information (PubMed) and the electronic library Scientific Electronic Library Online (SciELO). The search was carried out of articles published between 2008 and 2018, in Portuguese and / or English. Development: The efficacy of the vaccine is based on the immune response of the host against an antigen can induce a memory T cell response over time. There are several reports of cases of autoimmune diseases following vaccination, however, due to the limited number of cases, different classifications of the symptoms and the disease latency period, all attempts at anti-vaccination studies have so far failed to achieve a connection plausible. Conclusion: Therefore, the population should seek information from reliable sources and from government agencies. In addition, health professionals, education managers and the media should suggest an important strategy for maintaining vaccine coverage and addressing vaccine refusal ethically.Introdução: O processo de vacinação enfrenta, desde a sua implementação, questionamentos e críticas acerca de sua efetividade, funcionamento imunológico e resultados. À partir disso, mesmo sendo uma política pública eficiente no combate de doenças infectocontagiosas, tem-se a consciência da não realização da mesma, o que prejudica na prevenção da morbimortalidade das doenças infectocontagiosas. Métodos: Revisão bibliográfica utilizando como bases de dados o Google acadêmico, Medline, Center for Biotechnology Information (PubMed) e na biblioteca eletrônica Scientific Eletronic Library Online (SciELO). A busca foi realizada em artigos publicados no período de 2008 a 2018, em português e/ou inglês. Desenvolvimento: A eficácia da vacina baseia-se na resposta imune do hospedeiro contra um antígeno que possa induzir uma resposta da célula T de memória ao longo do tempo. Existem vários relatos de casos de doenças autoimunes após a vacinação; no entanto, devido ao número limitado de casos, as diferentes classificações dos sintomas e ao período de latência das doenças, todas as tentativas de estudos antivacinação, até o momento, não conseguiram uma correlação plausível. Conclusão: Portanto, a população deve buscar informações em fontes confiáveis e em órgãos governamentais. Além disso, profissionais da saúde, gestores da educação e a mídia devem sugerir estratégias importantes para manter as coberturas vacinais e abordar a recusa vacinal de forma ética

The anti-inflammatory and antitumor effects of medicinal plants: Arctium lappa, Solanum torvum and Lobelia inflata

Introduction: Medicinal plants have been used since antiquity to treat illnesses and injuries. Considering their global use, many natural products have been investigated with the aim to get new drugs. Methods: The search was based on relevant articles indexed in PubMed, Scielo and Scopus. The search terms used were: medicinal plants, Arctium lappa, Solanum torvum, Lobelia inflata, anti-inflammatory effects, antimicrobial activity and antitumor effects. Development: Arctium lappa leads to the inhibition of nitric oxide synthase (iNOS) expression and nitric oxide (NO) production and inhibits the growth of some tumor cell lines. Solanum torvum can promote inhibition of inflammatory mediators release, and reduces the melanoma formation. Lobelia inflata can reduce the number of white blood cells, the TNF-α and IL-6 levels and the melanoma growth. Conclusion: The active principles present in these medicinal plants, including flavonoids and other phenolic compounds with antioxidant activity, can scavenge free radicals and therefore be effective against tumors, such as melanoma and skin cancer

Nets desempenham papel importante nas doenças crônicas: uma revisão de literatura

Introduction: Neutrophil extracellular traps (NETs) are the result of extrusion of various components by neutrophils, and act to eradicate the pathogens present, as well as providing mechanical barrier, thus preventing the spread of microorganisms. Chronic diseases have been found to have a significant influence on these NETs, which by specific mechanisms produce negative effects on pathogenesis. Methods: Bibliographic review using Google Scholar, Medline, Center for Biotechnology Information (PubMed) and Scientific Electronic Library Online (SciELO) as databases. The search was performed in articles published from 2000 to 2019, in Portuguese and/or English languages. Development: It was evidenced that each disease in question is aggravated by NETs in a different way, either by inflammatory exacerbation or physical barrier, and the participation of pathogens in the neutrophil stimulation inducing process can be considered. Conclusion: Understanding the process of formation of NETs, ie, NETose, may help future findings that could contribute to mitigate the injury caused by the neutrophil mechanism, which involves multiple cytokines, granules and DNA molecules.extrusão de diversos componentes pelos neutrófilos, e agem com o intuito de erradicar os patógenos presentes, além de propiciar barreira mecânica, evitando assim, a disseminação de microrganismos. Foi descoberto que doenças crônicas possuem influência significativa dessas NETs, que por mecanismos específicos, produzem efeitos negativos sobre a patogênese. Métodos: Revisão bibliográfica utilizando como bases de dados o Google acadêmico, Medline, Center for Biotechnology Information (PubMed) e na biblioteca eletrônica Scientific Eletronic Library Online (SciELO). A busca foi realizada em artigos publicados no período de 2000 a 2019 em português e/ou inglês. Desenvolvimento: Foi evidenciado que cada doença em questão é agravada pelas NETs de uma maneira distinta, seja pela exacerbação inflamatória ou pela barreira física, e ainda pode-se considerar a participação de patógenos no processo indutor de estimulação dos neutrófilos. Conclusão: O entendimento do processo de formação das NETs, ou seja, a NETose, pode auxiliar em futuros achados que poderiam contribuir para amenizar a injúria causada pelo mecanismo dos neutrófilos, que envolve múltiplas citocinas, grânulos e moléculas de DNA

Partial purification and functional characterization of Ts19 Frag-I, a novel toxin from Tityus serrulatus scorpion venom

Abstract\ud \ud Background\ud The yellow scorpion Tityus serrulatus (Ts) is responsible for the highest number of accidents and the most severe scorpion envenoming in Brazil. Although its venom has been studied since the 1950s, it presents a number of orphan peptides that have not been studied so far. The objective of our research was to isolate and identify the components present in the fractions VIIIA and VIIIB of Ts venom, in order to search for a novel toxin. The major isolated toxins were further investigated for macrophage modulation.\ud \ud \ud Methods\ud The fractions VIIIA and VIIIB, obtained from Ts venom cation exchange chromatography, were rechromatographed on a C18 column (4.6 × 250 mm) followed by a reversed-phase chromatography using another C18 column (2.1 × 250 mm). The main eluted peaks were analyzed by MALDI-TOF and Edman’s degradation and tested on macrophages.\ud \ud \ud Results\ud The previously described toxins Ts2, Ts3-KS, Ts4, Ts8, Ts8 propeptide, Ts19 Frag-II and the novel peptide Ts19 Frag-I were isolated from the fractions VIIIA and VIIIB. Ts19 Frag-I, presenting 58 amino acid residues, a mass of 6,575 Da and a theoretical pI of 8.57, shares high sequence identity with potassium channel toxins (KTx). The toxins Ts4, Ts3-KS and the partially purified Ts19 Frag-I did not produce cytotoxic effects on macrophage murine cells line (J774.1). On the other hand, Ts19 Frag-I induced the release of nitric oxide (NO) by macrophages, while Ts4 and Ts3-KS did not affect the NO production at the tested concentration (50 μg/mL). At the same concentration, Ts19 Frag-I and Ts3-KS increased the production of interleukin-6 (IL-6). Ts19 Frag-I and Ts4 did not induce the release of IL-10, IL-1β or tumor necrosis factor-α by macrophage cells using the tested concentration (50 μg/mL).\ud \ud \ud Conclusions\ud We partially purified and determined the complete sequence and chemical/physical parameters of a new β-KTx, denominated Ts19 Frag-I. The toxins Ts4, Ts3-KS and Ts19 Frag-I showed no cytotoxicity toward macrophages and induced IL-6 release. Ts19 Frag-I also induced the release of NO, suggesting a pro-inflammatory activity.This study received financial support from the from the State of São Paulo\ud Research Foundation (FAPESP – scholarship to FAC n. 2012/13590-8 and\ud MBP n. 2012/12954-6), Coordination for the Improvement of Higher\ud Education Personnel (CAPES – scholarship to PCL), National Council for\ud Scientific and Technological Development (CNPq – grant n. 303689/2013-7)\ud and the Support Nucleus for Research on Animal Toxins (NAP-TOXAN-USP,\ud grant n. 12-125432.1.3). The authors would like to thank Prof. Dr. Norberto\ud Peporine Lopes for providing the MALDI-TOF mass spectrometer used in this\ud study. The authors also acknowledge the biologist Luiz Henrique Anzaloni Pedrosa\ud for extracting the scorpion venom and Iara Aimê Cardoso for technical assistance.\ud Thanks are also due to the Center for the Study of Venoms and Venomous\ud Animals (CEVAP) of UNESP for enabling the publication of this special collection\ud (CNPq process 469660/2014-7)

Paradoxical Effect of LTB4 on the Regulation of Stress-Induced Corticosterone Production

Depression is a mental illness with a complex and multifactorial etiology, which has been associated with stress and inflammation. Infections, autoimmune diseases, envenomation, and trauma induce an inflammatory response that is characterized by increasing levels of circulating cytokines (e.g., IL-1β) and lipid mediators [e.g., PGE2 and leukotrienes B4 (LTB4)]. Recently, we showed that LTB4 production by the 5-lipoxygenase (5-LO) pathway regulates IL-1β and PGE2 release, reducing tissue damage in a model of sterile inflammation. Since IL-1β and PGE2 increase in serum of stressed patients and potentially trigger depression, we used an animal model of chronic unpredictable stress (CUS) to investigate the potential impact of LTB4 over depression-like symptoms. At basal conditions, 5-LO deficiency (Alox5−/−) reduces the preference for sucrose, while inducing a higher immobilization time on the tail suspension test when compared 129sv. Moreover, Alox5−/− mice present increased caspase-1 expression and elevated levels of IL-1β, IL-17 and PGE2 in the spleen, with increasing corticosterone levels in the frontal cortex but reducing systemic levels. Compared to 129sv mice, CUS induced higher levels of systemic, frontal cortex and hippocampal corticosterone, and also reduced sucrose preference, increased levels of splenic IL-1β, IL-17 and PGE2 and reduced levels of LTB4. Interestingly, CUS exposure did not alter the reduced sucrose preference shown by Alox5−/− mice but greatly enhanced splenic PGE2 production. Compared to Alox5−/− mice at basal conditions, CUS exposure also increased levels of systemic corticosterone, which remained lower than those of CUS-129sv animals. We also observed that treatment with LTB4 decreased caspase-1 expression and systemic levels of corticosterone in CUS-Alox5−/− mice but there was no significant impact on the reduced sucrose preference. Our results demonstrate that LTB4 controls the hypothalamic-pituitary-adrenal (HPA) axis by regulating levels of systemic corticosterone associated with the repression of caspase-1 expression and production of inflammatory mediators. One limitation of our study is that 129sv and Alox5−/− mice were not littermates, not sharing, therefore, the same intra-uterine and preweaning environment. Even so, taken together our results indicate that 5-LO activity is critical for the regulation of stress-induced symptoms, suggesting that the Alox5−/− mouse could be a natural model of corticosterone-independent reduced reward sensitivity

Monitoramento no âmbito hospitalar: avaliação da carga microbiana

Introduction: Hospital infections (HI) constitute a serious public health problem, which may occur during hospitalization or after discharge. It is undeniable that environmental contamination, involving important microorganisms (Staphylococcus aureus, Enterococcus, Acinetobacter spp., Clostridium difficile, among others) represents a risk of transmission between patients and professionals.. Objective: Given the importance of microbial control in HI prevention, the present study evaluated the microbial load of health professionals of a public hospital in the interior of the state of São Paulo, through the hand washing and dressing procedure, as well as the hospital environment. Methods: To perform the data collection, the Termo de Consentimento Livre e Esclarecido (TCLE) was applied. After the clarification of the TCLE, for the microbiological collections of the hands and clothes of the health professionals, laminous crops were used. And for the evaluation of the microbial load of the air, the sedimentation plate technique was used. Results: We observed the efficiency of the hand hygiene technique, since partial or total reduction of the microbial load occurred. As for clothing collections, the results are increasing numerically at the expense of the workday, in addition to presenting characteristics intrinsic to the activity developed by the professional. The results obtained through the sedimentation plates indicate that areas of lower flow of people and / or controlled access, such as beds, have a reduced total microbial load. Conclusion: The presence of microorganisms in the hospital environment is inevitable, however, prophylactic measures such as hand hygiene, and disinfection of surfaces and the correct use of clothes or periodic exchanges of the same contribute positively to the prevention of infections via the exogenous route.Introdução: As infecções hospitalares (IH) constituem um grave problema de saúde pública, podendo ocorrer durante o período de internação ou após a alta. É incontestável que a contaminação ambiental, envolvendo importantes microrganismos (Staphylococcus aureus, Enterococcus, Acinetobacter spp., Clostridium difficile entre outros) representa risco de transmissão entre pacientes e profissionais. Objetivo: Dado a importância do controle microbiano na prevenção de IH, o presente estudo avaliou a carga microbiana dos profissionais da saúde de um hospital público do interior do estado de São Paulo, através do procedimento de lavagem das mãos e das vestimentas, bem como do ambiente hospitalar. Métodos: Para realização das coletas foi aplicado o Termo de Consentimento Livre e Esclarecido (TCLE). Após o esclarecimento do TCLE, foram utilizados laminocultivos para as coletas microbiológicas das mãos e vestimentas dos profissionais da saúde. Para avaliação da carga microbiana do ar, foi utilizado a técnica da placa de sedimentação. Resultados: Nós observamos a eficiência da técnica de higienização das mãos, visto que ocorreu redução parcial ou total da carga microbiana. Já para as coletas das vestimentas, os resultados apresentam-se crescente numericamente em detrimento do decorrer da jornada de trabalho, além de apresentarem características intrínsecas à própria atividade desenvolvida pelo profissional. Os resultados obtidos através das placas de sedimentação indicam que áreas de menor fluxo de pessoas e/ou de acesso controlado, como os leitos, possuem a carga microbiana total reduzida. Conclusão: A presença de microrganismos no ambiente hospitalar é inevitável, entretanto, medidas profiláticas como a higienização das mãos, desinfecção de superfícies e a correta utilização das vestimentas ou troca periódica das mesmas contribuem positivamente na prevenção de infecções pela via exógena

Atheroregressive Potential of the Treatment with a Chimeric Monoclonal Antibody against Sulfated Glycosaminoglycans on Pre-existing Lesions in Apolipoprotein E-Deficient Mice

The retention of lipoprotein particles in the intima, in particular to glycosaminoglycan side chains of proteoglycans, is a critical step in atherosclerosis initiation. Administration of chP3R99, a chimeric mouse/human monoclonal antibody inducing an anti-idiotypic network response against glycosaminoglycans was previously shown to prevent atherosclerotic lesion progression, yet its effect in the late-stage progression of lesions remains unknown. This study investigated the effect of chP3R99 at a late stage of disease development in apolipoprotein E-deficient mice and the vascular mechanisms involved. Male apolipoprotein E-deficient mice were fed a high-fat high-cholesterol diet from 4 to 19 weeks old, at which time mice were fed normal chow and 5 doses of chP3R99 (50 μg) or isotype-matched IgG (hR3) were administered subcutaneously weekly for the first 3 administrations, then at weeks 24 and 26 before sacrifice (week 28). Lesions progression was reduced by 88% in treated mice with no change in total plasma cholesterol levels, yet with increased sera reactivity to chP3R99 idiotype and heparin, suggesting the induction of an anti-idiotype antibody cascade against glycosaminoglycans, which was likely related with the atheroprotective effect. chP3R99 treatment initiated regression in a significant number of mice. Circulating levels of interleukin-6 were reduced along with a striking diminution of inflammatory cell accumulation in the vessel wall, and of VCAM-1 labeling in vivo. The ratio of IL-10/iNOS gene expression in aortas increased in chP3R99-treated mice. In conclusion, our results show that treatment with chP3R99 reduces vascular inflammatory burden and halts lesion progression with potential for regression in the late phase of the disease in atherosclerotic mice, and support the therapeutic intervention against glycosaminoglycans as a novel strategy to reverse atherosclerosis

Green Propolis Compounds (Baccharin and p-Coumaric Acid) Show Beneficial Effects in Mice for Melanoma Induced by B16f10

Background: Cutaneous melanoma is the most aggressive form of skin cancer, with the worst prognosis, and it affects a younger population than most cancers. The high metastatic index, in more advanced stages, and the high aggressiveness decrease the effectiveness of currently used therapies, such as surgical removal, radiotherapy, cryotherapy, and chemotherapy, used alone or in combination. Based on these disadvantages, research focused on alternative medicine offers great potential for therapeutic innovation. Medicinal plants represent a remarkable source of compounds for the treatment of various diseases. Methods: In this study, we investigated the tumoral behavior of melanoma under treatment with the compounds baccharin and p-coumaric acid, extracted from green propolis, in mice inoculated with B16F10 cells for 26 days. Results: A significant modulation in the number of inflammatory cells recruited to the tumor region and blood in the groups treated with the compounds was observed. In addition, a significant reduction in the amount of blood vessels and mitosis in the neoplastic area was noticed. Conclusions: Through our research, we confirmed that baccharin and coumaric acid, isolated substances from Brazilian green propolis, have a promising anticarcinogenic potential to be explored for the development of new antitumor agents, adhering to the trend of drugs with greater tolerance and biological effectiveness