CXCL5-mediated recruitment of neutrophils into the peritoneal cavity of Gdf15-deficient mice protects against abdominal sepsis.

Sepsis is a life-threatening organ dysfunction condition caused by a dysregulated host response to an infection. Here we report that the circulating levels of growth and differentiation factor-15 (GDF15) are strongly increased in septic shock patients and correlate with mortality. In mice, we find that peptidoglycan is a potent ligand that signals through the TLR2-Myd88 axis for the secretion of GDF15, and that Gdf15-deficient mice are protected against abdominal sepsis due to increased chemokine CXC ligand 5 (CXCL5)-mediated recruitment of neutrophils into the peritoneum, leading to better local bacterial control. Our results identify GDF15 as a potential target to improve sepsis treatment. Its inhibition should increase neutrophil recruitment to the site of infection and consequently lead to better pathogen control and clearance.

DNA damage independent inhibition of NF-κB transcription by anthracyclines.

Anthracyclines are among the most used and effective anticancer drugs. Their activity has been attributed to DNA double-strand breaks resulting from topoisomerase II poisoning and to eviction of histones from select sites in the genome. Here, we show that the extensively used anthracyclines Doxorubicin, Daunorubicin, and Epirubicin decrease the transcription of nuclear factor kappa B (NF-κB)-dependent gene targets, but not interferon-responsive genes in primary mouse (Mus musculus) macrophages. Using an NMR-based structural approach, we demonstrate that anthracyclines disturb the complexes formed between the NF-κB subunit RelA and its DNA-binding sites. The anthracycline variants Aclarubicin, Doxorubicinone, and the newly developed Dimethyl-doxorubicin, which share anticancer properties with the other anthracyclines but do not induce DNA damage, also suppressed inflammation, thus uncoupling DNA damage from the effects on inflammation. These findings have implications for anticancer therapy and for the development of novel anti-inflammatory drugs with limited side effects for life-threatening conditions such as sepsis.

Evaluation of Different Methods for Cultivating Gluconacetobacter hansenii for Bacterial Cellulose and Montmorillonite Biocomposite Production: Wound-Dressing Applications.

Bacterial cellulose (BC) has received considerable attention due to its unique properties, including an ultrafine network structure with high purity, mechanical strength, inherent biodegradability, biocompatibility, high water-holding capacity and high crystallinity. These properties allow BC to be used in biomedical and industrial applications, such as medical product. This research investigated the production of BC by Gluconacetobacter hansenii ATCC 23769 using different carbon sources (glucose, mannitol, sucrose and xylose) at two different concentrations (25 and 50 g∙L-1). The BC produced was used to develop a biocomposite with montmorillonite (MMT), a clay mineral that possesses interesting characteristics for enhancing BC physical-chemical properties, at 0.5, 1, 2 and 3% concentrations. The resulting biocomposites were characterized in terms of their physical and barrier properties, morphologies, water-uptake capacities, and thermal stabilities. Our results show that bacteria presented higher BC yields in media with higher glucose concentrations (50 g∙L-1) after a 14-day incubation period. Additionally, the incorporation of MMT significantly improved the mechanical and thermal properties of the BC membranes. The degradation temperature of the composites was extended, and a decrease in the water holding capacity (WHC) and an improvement in the water release rate (WRR) were noted. Determining a cost-effective medium for the production of BC and the characterization of the produced composites are extremely important for the biomedical applications of BC, such as in wound dressing materials.

Students' knowledge of metabolic syndrome after educational intervention.

OBJECTIVE: To analyze the knowledge of children and adolescents about risk factors for metabolic syndrome (MS) before and after educational interventions. METHOD: A quasi-experimental, comparative, prospective and intervention study conducted in 2015 and 2016 with 43 students in the city of Picos (state of Piauí-PI). Five health education meetings were held. For the knowledge analysis, was applied a questionnaire at three different moments. RESULTS: Participants' mean age was 12.6 years (± 2.1), of which 60.5% were female. The 'More than good' level of knowledge went from 20.9% to 55.8% after interventions. When evaluated late, students maintained a higher level of knowledge compared to before the interventions. Most said they were able to change their lifestyle after educational meetings. CONCLUSION: The educational intervention promoted increase of knowledge and stimulated changes in attitudes related to risk factors associated with MS.

[Evaluation of different preservation solutions utilized in the machine perfusion of kidneys retrieved under cardiac arrest. An experimental study]. / Avaliação da eficácia de diferentes soluções de preservaçao utilizadas na perfusão pulsátil de rins colhidos em circunstâncias de paragem cardíaca. Estudo experimental.

Considering the special demands of organs retrieved after cardiac death, the solution that should optimize and preserve the best early graft function and long-term graft survival remains to be determined. Recently, polyethylene glycol (PEG), high-molecular weight colloid, due to its protective effect against ischemia and reperfusion injury, has been proposed to be added to different preservation solutions for cold storage. Celsior formulation was targeted to fulfil the principles of organ preservation. The aim of our study was to evaluate Celsior plus PEG as a machine perfusion solution for kidneys retrieved after cardiac death. Landrace pigs were killed by lethal injection. Kidneys were submitted to warm and cold ischemic injuries mimicking the injury suffered by donation after cardiac death and were machine perfused with Celsior, Celsior plus 30 mg/l of PEG 20,000 Da and Belzer MPS. We demonstrate that when kidneys, submitted to warm and cold ischemia injury, were machine perfused with Celsior plus PEG the IRR were lower and the renal flow rates were higher in comparison with Celsior alone or Belzer MPS in the same conditions. This study provides the first evidence that Celsior plus PEG is an effective solution for machine perfusion of kidneys retrieved after cardiac death.

Sepsis: the need for tolerance not complacency.

Sepsis is a life-threatening condition that arises as a systemic inflammatory response syndrome to an infection. Its uncontrolled progression can in frequent cases lead to multiple organ failure, which is still associated with high mortality rates. Modern antibiotics made clear that the infection is only an initiating, and not always necessary, event of this syndrome as many patients with sepsis die despite effective eradication of the inciting pathogen. This observation critically contributed to a paradigm shift that focused the pathogenesis of sepsis on the host and not on the pathogen. However, therapeutic strategies based on the inhibition of proinflammatory critical mediators of sepsis or immunostimulation have so far failed to improve sepsis outcome and, therefore, this condition urgently needs transformative therapeutic ideas and strategies. Here we argue that the induction of tolerance, a defence strategy that minimises the impact of an infection on organ function without directly affecting the pathogen burden, is perhaps the missing but essential element to add to the current components of sepsis care and treatment.

Additional peculiarities of medical devices that should be considered in their development process.

Medical devices are peculiar products: their definition varies from country to country, they are used to treat diseases and they are different from pharmaceuticals. In 2012, the authors began to describe the complex and demanding environment of the medical device industry. In this article, the authors' previous research is extended with additional peculiarities of medical devices such as recall, pricing and adoption factors.

Students' knowledge of metabolic syndrome after educational intervention / Conocimiento de estudiantes sobre síndrome metabólico tras intervención educative / Conhecimento de estudantes sobre síndrome metabólica após intervenção educativa

ABSTRACT Objective: To analyze the knowledge of children and adolescents about risk factors for metabolic syndrome (MS) before and after educational interventions. Method: A quasi-experimental, comparative, prospective and intervention study conducted in 2015 and 2016 with 43 students in the city of Picos (state of Piauí-PI). Five health education meetings were held. For the knowledge analysis, was applied a questionnaire at three different moments. Results: Participants' mean age was 12.6 years (± 2.1), of which 60.5% were female. The 'More than good' level of knowledge went from 20.9% to 55.8% after interventions. When evaluated late, students maintained a higher level of knowledge compared to before the interventions. Most said they were able to change their lifestyle after educational meetings. Conclusion: The educational intervention promoted increase of knowledge and stimulated changes in attitudes related to risk factors associated with MS.

RESUMEN Objetivo: Analizar el conocimiento de niños y adolescentes acerca de los factores de riesgo para el síndrome metabólico (SM) antes y después de intervenciones educativas. Método: Estudio casi experimental, comparativo, prospectivo y de intervención, realizado en 2015 y 2016, con 43 estudiantes en Picos, estado del Piauí (PI). Se realizaron cinco encuentros de educación en salud. Para el análisis del conocimiento, se utilizó un cuestionario aplicado en tres momentos distintos. Resultados: Los participantes tenían en promedio 12,6 años (± 2,1), siendo el 60,5% del sexo femenino. El nivel de conocimiento 'Más que bueno' pasó del 20,9% al 55,8% después de las intervenciones. Cuando evaluados tardíamente, los estudiantes mantuvieron un nivel de conocimiento mayor comparado con antes de las intervenciones. La mayoría refirió haber logrado cambiar actitudes en cuanto al estilo de vida después de los encuentros. Conclusión: La intervención educativa promovió aumento del conocimiento y estímulo a los cambios de actitudes acerca de los factores de riesgo asociado al SM.

RESUMO Objetivo: Analisar o conhecimento de crianças e adolescentes acerca dos fatores de risco para síndrome metabólica (SM) antes e após intervenções educativas. Método: Estudo do tipo quase experimental, comparativo, prospectivo e de intervenção, realizado em 2015 e 2016, com 43 estudantes em Picos-PI. Realizaram-se cinco encontros de educação em saúde; para a análise do conhecimento, foi utilizado um questionário aplicado em três momentos distintos. Resultados: Os participantes tinham em média 12,6 anos (± 2,1), sendo 60,5% do sexo feminino. O nível de conhecimento "Mais que bom" passou de 20,9% para 55,8% após as intervenções. Quando avaliados tardiamente, os estudantes mantiveram um nível de conhecimento maior quando comparados antes das intervenções. A maioria referiu ter conseguido mudar atitudes quanto ao estilo de vida após os encontros. Conclusão: A intervenção educativa promoveu aumento do conhecimento e estímulo às mudanças de atitudes acerca dos fatores de risco associado à SM.

Comparison of phylogeny, venom composition and neutralization by antivenom in diverse species of bothrops complex.

In Latin America, Bothrops snakes account for most snake bites in humans, and the recommended treatment is administration of multispecific Bothrops antivenom (SAB--soro antibotrópico). However, Bothrops snakes are very diverse with regard to their venom composition, which raises the issue of which venoms should be used as immunizing antigens for the production of pan-specific Bothrops antivenoms. In this study, we simultaneously compared the composition and reactivity with SAB of venoms collected from six species of snakes, distributed in pairs from three distinct phylogenetic clades: Bothrops, Bothropoides and Rhinocerophis. We also evaluated the neutralization of Bothrops atrox venom, which is the species responsible for most snake bites in the Amazon region, but not included in the immunization antigen mixture used to produce SAB. Using mass spectrometric and chromatographic approaches, we observed a lack of similarity in protein composition between the venoms from closely related snakes and a high similarity between the venoms of phylogenetically more distant snakes, suggesting little connection between taxonomic position and venom composition. P-III snake venom metalloproteinases (SVMPs) are the most antigenic toxins in the venoms of snakes from the Bothrops complex, whereas class P-I SVMPs, snake venom serine proteinases and phospholipases A2 reacted with antibodies in lower levels. Low molecular size toxins, such as disintegrins and bradykinin-potentiating peptides, were poorly antigenic. Toxins from the same protein family showed antigenic cross-reactivity among venoms from different species; SAB was efficient in neutralizing the B. atrox venom major toxins. Thus, we suggest that it is possible to obtain pan-specific effective antivenoms for Bothrops envenomations through immunization with venoms from only a few species of snakes, if these venoms contain protein classes that are representative of all species to which the antivenom is targeted.

Modeling of the medical device development process.

Models are abstract representations of reality that are built, analyzed and manipulated to augment the understanding of that reality. They can either be mental or codified but, in both cases, they contribute to good decision-making by ensuring that the right people use the right information at the right time. In order to reduce time to market, companies commonly adopt product development processes (PDP) and the medical device sector is no different. However, one can question which is the most adequate PDP model to follow or what information that model should contain, or even how the information should be represented. Here, the authors review the existing PDP models dedicated to medical devices. The model's representation and usability are also debated in order to assist developers to select the most suitable model and adapt it to their needs and reality.

Medical device specificities: opportunities for a dedicated product development methodology.

The medical sector, similarly to other industries such as the aviation industry, has to comply with multiple regulations, guidelines and standards. In addition, there are multiple definitions for the expression 'medical device', and before entering the market, manufacturers must demonstrate their product's safety and effectiveness. In such a complex and demanding environment, it is crucial to know the particularities surrounding the product being developed in order to minimize the chances of a commercial flop. Thus, in this paper, medical device specificities are identified, and the most relevant legislation is reviewed providing the foundations for a dedicated product development methodology.

Estilo de vida e fatores associados entre estudantes universitários / Lifestyle and associated factors between university students

Objetivo: estimar a proporção de alcoolismo, tabagismo, sedentarismo entre universitários e fatores associados. Método: estudo descritivo, de abordagem quantitativa, realizado com 550 estudantes de uma Instituição Pública de Ensino Superior. Os dados foram coletados a partir de formulário, armazenados no SPSS20.0, analisados e apresentados em tabelas. Resultados: verificou-se que o sexo masculino (p=0,000) e os cursos das áreas de Humanas e Exatas (p=0,009) apresentam associação significante com o consumo de álcool danoso ao corpo humano. Em relação à atividade física, observou-se maior proporção de sedentarismo no sexo feminino (p=0,000) e nos universitários pertencentes às classes econômicas C-D-E (p=0,028). Conclusão: são evidentes os riscos relacionados ao estilo de vida ao qual estão expostos os universitários. Daí a importância do desenvolvimento de medidas educativas que estimulem a adoção hábitos saudáveis de vida.

Obtenção e atividade biológica de uma nova disintegrina recombinante do veneno de Bothrops neuwiedi, Dis-BnMPIIx / Obtention and biological activity of a new recombinant disintegrin from Bothrops neuwiedi venom, Dis-BnMPIIx

Metalloproteinases from snake venoms are highly versatile enzymes, responsible for most of the symptoms of envenoming. Its action is related to the proteolysis of extracellular matrix components, activation or hydrolysis of components of the coagulation cascade and fibrinolysis, platelet aggregation inhibition, induction of local and systemic bleeding, pro-inflammatory activity, among others. This broad variety of biological activities is related to the structural diversity of this family of proteins. The SVMPs are divided into classes and subclasses of PI to PIII according to the organization of their domains. The SVMPs class PI are composed only by the catalytic domain, and in the class PIII, besides catalytic domain, SVMPs have disintegrin-like and cysteine-rich domains. The SVMPs class PII are composed of catalytic and disintegrin domain or only disintegrin that generally present the RGD tripeptide. For a better understanding of the structural and functional diversity of SVMPs, cDNAs from venom gland Bothrops neuwiedi encoding SVMPs were sequenced. SVMPs distinct from those described in venoms of other species were found. The BnMPIIx is a SVMP class PII showing a peculiar sequence with the fusion of a catalytic domain typical of procoagulant class P-III SVMPs with an RGDdisintegrin domain of SVMPs classP-II, with the inclusion of cysteines at positions undescribed previously. The aim of this work was to obtain of BnMPIIx and its disintegrin domain in recombinant form using vectors directing to the E. coli periplasm (pET20) or vectors that produce the recombinant protein in fusion with chaperones (pET32 and pSMT3). For this, the cDNAs encoding BnMPIIx and its disintegrin domain were amplified by PCR, digested with endonucleases BamHI and HindIII and cloned into pET20 vector. The disintegrin domain was also cloned into pET32 and pSMT3 vectors that express proteins with chaperones thioredoxin (Trx) and SUMO, respectively. The sequencing of the inserts did not show any mutations introduced during the cloning procedures. These vectors were transformed into the E. coli BL21 (DE3) or C43 (DE3) strains, and expression was evaluated at two temperatures (30°C and 37°C). To test platelet aggregation, human blood was collected in a 3,8% sodium citrate (1:9) from healthy donors. The mixture was centrifuged and the platelet-rich plasma (PRP) was submitted to an ADP-induced platelet aggregation assay. The analysis by SDS-PAGE showed that both BnMPIIx as its disintegrin domain were obtained by pET20 partially soluble in both temperatures (30°C and 37°C), resulting in inclusion bodies. Using the vectors pET32 and pSMT3 the disintegrin domain was expressed in a soluble form and was called Dis-Trx or Dis-SUMO respectively. However, the fusion proteins did not show biological activity. After cleavage of Trx it was not possible to isolate the disintegrin to validate the biological activity of Trx-Dis, but after cleavage of SUMO, the isolated disintegrin completely inhibited platelet aggregation in a concentration of approximately 2 µM. After several attempts using different expression vectors, we obtained a new recombinant disintegrin from Bothrops neuwiedi venom in pSMT3 vector, with preserved biological activity.This molecule may contribute to the study of hemostasis and cancer.

As metaloproteinases de venenos de serpentes (SVMPs) são enzimas altamente versáteis, responsáveis por grande parte dos sintomas do envenenamento. Sua ação está relacionada com a proteólise dos componentes da matriz extracelular, ativação ou hidrólise de componentes da cascata de coagulação e fibrinólise, inibição da agregação plaquetária, indução de hemorragia local e sistêmica, atividade pró-inflamatória, entre outras. Essa ampla gama de atividades biológicas está relacionada com a diversidade estrutural dessa família de proteínas. As SVMPs são divididas em classes e subclasses de PI a PIII de acordo com a organização dos seus domínios. As SVMPs de classe PI são compostas apenas pelo domínio catalítico, já as PIII possuem domínio catalítico, tipo disintegrina e rico em cisteína. As SVMPs PII, possuí domínio catalítico e disintegrina ou apenas de disintegrina livre que geralmente apresenta o tripeptídeo RGD. Para melhor entender a diversidade estrutural e funcional das SVMPs, foram sequenciados cDNAs da glândula de veneno Bothrops neuwiedi que codificam SVMPs. Foram encontradas novas SVMPs, distintas das descritas em venenos de outras espécies. A BnMPIIx, uma SVMP de classe PII,apresentou uma sequência bastante peculiar com a fusão de um domínio catalítico similar ao de SVMPs pró-coagulantes da classe P-III com um domínio RGD-disintegrina, de SVMPs classe P-II, contendo cisteínas em posições não descritas anteriormente. O objetivo deste trabalho foi obter a BnMPIIx e seu domínio disintegrina na forma recombinante através de vetores de E. coli que direcionam para o periplasma (pET20) ou vetores que proporcionam a fusão com chaperonas (pET32 e pSMT3). Para isto, o cDNA da BnMPIIx e de seu dominínio disintegrina foram amplificados por PCR, digeridos com as endonucleases BamHI e HindIII e clonados em vetor pET20. O domínio disintegrina também foi clonado nos vetores pET32 e pSMT3 que expressam as proteínas com as chaperonas Tioredoxina (Trx) e SUMO, respectivamente. O sequenciamento de todos os insertos não apresentou mutações. Estes vetores foram transformados nas linhagens de E. coli BL21 (DE3) ou C43 (DE3) e a expressão avaliada em duas temperaturas (30ºC e 37ºC). Para o ensaio de inibição da agregação plaquetária, foi coletado sangue humano de doadores voluntários saudáveis em 3,8% de citrato de sódio (1:9). O sangue foi centrifugado e o plasma rico em plaquetas (PRP) submetido à agregação plaquetária induzida por ADP. A análise por SDS-PAGE mostrou que tanto a BnMPIIx quanto seu domínio disintegrina foram obtidos parcialmente solúveis em ambas as temperaturas (30ºC e 37ºC), gerando corpos de inclusão em vetor pET20. Já a disintegrina foi obtida com sucesso nos vetores pET32 e pSMT3 e foram denominadas Dis-Trx e Dis-SUMO respectivamente, no entanto não apresentaram atividade biológica. Após a clivagem da Trx, não foi possível isolar a disintegrina para validar a atividade biológica da Dis-Trx, já após a clivagem da SUMO a disintegrina inibiu completamente a agregação plaquetária em concentrações da ordem de 2 µM. Após diversas tentativas de clonagem usando diferentes vetores, nós obtivemos uma nova disintegrina recombinante de Bothrops neuwiedi em vetor pSMT3 com atividade biológica preservada, podendo esta molécula contribuir para os estudos de hemostasia e câncer.