The Therapeutic Treatment with the GAG-Binding Chemokine Fragment CXCL9(74-103) Attenuates Neutrophilic Inflammation and Lung Dysfunction during Klebsiella pneumoniae Infection in Mice.

Klebsiella pneumoniae is an important pathogen associated with hospital-acquired pneumonia (HAP). Bacterial pneumonia is characterized by a harmful inflammatory response with a massive influx of neutrophils, production of cytokines and chemokines, and consequent tissue damage and dysfunction. Targeted therapies to block neutrophil migration to avoid tissue damage while keeping the antimicrobial properties of tissue remains a challenge in the field. Here we tested the effect of the anti-inflammatory properties of the chemokine fragment CXCL9(74-103) in pneumonia induced by Klebsiella pneumoniae in mice. Mice were infected by intratracheal injection of Klebsiella pneumoniae and 6 h after infection were treated systemically with CXCL9(74-103). The recruitment of leukocytes, levels of cytokines and chemokines, colony-forming units (CFU), and lung function were evaluated. The treatment with CXCL9(74-103) decreased neutrophil migration to the airways and the production of the cytokine interleukin-1ß (IL-1ß) without affecting bacterial control. In addition, the therapeutic treatment improved lung function in infected mice. Our results indicated that the treatment with CXCL9(74-103) reduced inflammation and improved lung function in Klebsiella pneumoniae-induced pneumonia.

Pulmonary macrophages and their different roles in health and disease.

Macrophages are a heterogeneous population of myeloid cells with phenotype and function modulated according to the microenvironment in which they are found. The lung resident macrophages known as Alveolar Macrophages (AM) and Interstitial Macrophages (IM) are localized in two different compartments. During lung homeostasis, macrophages can remove inhaled particulates, cellular debris and contribute to some metabolic processes. Macrophages may assume a pro-inflammatory phenotype after being classically activated (M1) or anti-inflammatory when being alternatively activated (M2). M1 and M2 have different transcription profiles and act by eliminating bacteria, viruses and fungi from the host or repairing the damage triggered by inflammation, respectively. Nevertheless, macrophages also may contribute to lung damage during persistent inflammation or continuous exposure to antigens. In this review, we discuss the origin and function of pulmonary macrophages in the context of homeostasis, infectious and non-infectious lung diseases.

Shortened derivatives from native antimicrobial peptide LyeTx I: In vitro and in vivo biological activity assessment.

In the continuing search for novel antibiotics, antimicrobial peptides are promising molecules, due to different mechanisms of action compared to classic antibiotics and to their selectivity for interaction with microorganism cells rather than with mammalian cells. Previously, our research group has isolated the antimicrobial peptide LyeTx I from the venom of the spider Lycosa erythrognatha. Here, we proposed to synthesize three novel shortened derivatives from LyeTx I (LyeTx I mn; LyeTx I mnΔK; LyeTx I mnΔKAc) and to evaluate their toxicity and biological activity as potential antimicrobial agents. Peptides were synthetized by Fmoc strategy and circular dichroism analysis was performed, showing that the three novel shortened derivatives may present membranolytic activity, like the original LyeTx I, once they folded as an alpha helix in 2.2.2-trifluorethanol and sodium dodecyl sulfate. In vitro assays revealed that the shortened derivative LyeTx I mnΔK presents the best score between antimicrobial (↓ MIC) and hemolytic (↑ EC50) activities among the synthetized shortened derivatives, and LUHMES cell-based NeuriTox test showed that it is less neurotoxic than the original LyeTx I (EC50 [LyeTx I mnΔK] ⋙ EC50 [LyeTx I]). In vivo data, obtained in a mouse model of septic arthritis induced by Staphylococcus aureus, showed that LyeTx I mnΔK is able to reduce infection, as demonstrated by bacterial recovery assay (∼10-fold reduction) and scintigraphic imaging (less technetium-99m labeled-Ceftizoxime uptake by infectious site). Infection reduction led to inflammatory process and pain decreases, as shown by immune cells recruitment reduction and threshold nociception increment, when compared to positive control group. Therefore, among the three shortened peptide derivatives, LyeTx I mnΔK is the best candidate as antimicrobial agent, due to its smaller amino acid sequence and toxicity, and its greater biological activity.

Lipoxin A4 impairs effective bacterial control and potentiates joint inflammation and damage caused by Staphylococcus aureus infection.

Staphylococcus aureus is the main cause of septic arthritis in humans, a disease associated with high morbidity and mortality. Inflammation triggered in response to infection is fundamental to control bacterial growth but may cause permanent tissue damage. Here, we evaluated the role of Lipoxin A4 (LXA4 ) in S aureus-induced arthritis in mice. Septic arthritis was induced by S aureus injection into tibiofemoral joints. At different time points, we evaluated cell recruitment and bacterial load in the joint, the production of pro-inflammatory molecules, and LXA4 in inflamed tissue and analyzed joint damage and dysfunction. LXA4 was investigated using genetically modified mice or by pharmacological blockade of its synthesis and receptor. CD11c+ cells were evaluated in lymph nodes by confocal microscopy and flow cytometry and dendritic cell chemotaxis using the Boyden chamber. Absence or pharmacological blockade of 5-lipoxygenase (5-LO) reduced joint inflammation and dysfunction and was associated with better control of infection at 4 to 7 days after the infection. There was an increase in LXA4 in joints of S aureus-infected mice and administration of LXA4 reversed the phenotype in 5-LO-/- mice. Blockade or absence of the LXA4 receptor FPR2 has a phenotype similar to 5-LO-/- mice. Mechanistically, LXA4 appeared to control migration and function of dendritic cells, cells shown to be crucial for adequate protective responses in the model. Thus, after the first days of infection when symptoms become evident therapies that inhibit LXA4 synthesis or action could be useful for treatment of S aureus-induced arthritis.

Modos de ser e trabalhar de caminhoneira: estratégia de viver a vida / Ways of being and work of womantruck drive: strategy of living the life / Modos de ser y trabajar de camionera: estrategia de vivir la vida / Modes d'être et de travailler comme conductrice de camion: stratégie pour vivre la vie

O estudo destaca a estratégia de viver a vida de caminhoneira, suportado nas noções de trabalho imaterial e vida líquida. Caminhoneiras atuantes no transporte rodoviário de cargas de longo percurso e duração participaram da pesquisa exploratória por meio de entrevistas semiestruturadas. A análise do conteúdo apontou que a caminhoneira desempenha atividades que se aproximam da noção de trabalho imaterial e possibilitou estender a noção de estratégia tomada de Bauman à vida cotidiana. Seu dia a dia remete à vida líquida; remete à indistinção entre tempo/espaço de trabalho e vida; ao percurso e cenário que extrapolam a boleia; aos elementos culturais associados à estrada. Entre truques e traquejos, a estratégia de viver a vida concentra-se em reinícios indolores que demandam manter o jogo curto e o itinerário minimamente reconhecível. Para prosseguir na profissão, em perspectiva gradiente, a caminhoneira contempla facetas do turista, andarilho e jogador, conforme metáforas apresentadas por Bauman.

The study highlights the strategy of living the life of a female truck driver, based on the notions of immaterial labor and liquid life. Female truck driver engaged in long-distance and long-haul freight transportation participated in the exploratory research through semi-structured interviews. The analysis of the content indicated that the female truck driver performs activities that approach the notion of immaterial work and made possible to extend the notion of strategy taken from Bauman to daily life. Her daily life refers to liquid life; refers to the indistinction between time / space of work and life; the route and scenario that extrapolate the truck; cultural elements associated with the road. Between tricks and great experience, the strategy of living the life concentrates in painless restarts that demand to keep the game short and the itinerary minimally recognizable. To continue in the profession, in gradient perspective, the female trucker driver contemplates facets of the tourist, wanderer and player, according to metaphors presented by Bauman.

Este trabajo enfoca la estrategia de vivir la vida como camionera, basada en las nociones de trabajo inmaterial y vida líquida. Camioneras actuantes en el transporte de cargas en carretera de largo trayecto y tiempo participaron de la investigación de exploración por medio de entrevistas semiestructuradas. El análisis del contenido indicó que la camionera cumple actividades que se acercan a la noción de trabajo inmaterial y posibilitó la extensión de la noción de estrategia de Bauman a la vida cotidiana. Su jornada remete a la vida líquida; remete a la indistinción entre tiempo/espacio de trabajo y vida; al trayecto y escenario que extrapolan la cabina; a los elementos culturales relacionados a la carretera. Entre trucos y experiencias, la estrategia de vivir la vida está centrada en nuevos comienzos indoloros que demandan mantener el corto juego y la ruta mínimamente reconocible. Para seguir adelante en la profesión, en perspectiva gradiente, la camionera contempla facetas del turista, andador y jugador, según metáforas presentadas por Bauman.

L'étude met en évidence la stratégie de vivre la vie d'un conductrice de camion. Pour cela, on s'appuie sur les notions de travail immatériel et de vie liquide. Les conductrices de camions engagées dans le transport de marchandises sur des longues distances ont participé à la recherche exploratoire au moyen d'entretiens semi-structurés. L'analyse du contenu indique que le conductrice de camion exerce des activités qui abordent la notion de travail immatériel et a permit d'étendre la notion de stratégie prise de Bauman à la vie quotidienne. Sa vie quotidienne fait référence à la vie liquide; fait référence à la distinction entre temps / espace de travail et de vie; à la route et au scénario qui extrapole la cabine du camion; aux éléments culturels associés à la route. Entre astuces et compétences, la stratégie de vivre la vie se concentre sur des redémarrages sans douleur qui exigent de garder le jeu court et l'itinéraire minimalement reconnaissable. Pour exercer son métier, dans une perspective gradient, la conductrice de camion contemple les facettes du touriste, du voyageur et du joueur, selon les métaphores présentées par Bauman.

Regimes de flexibilização e sentidos do trabalho para docentes de ensino superior em instituições públicas e privadas / Flexible work arrangements and the meaning of work for teachers of higher education in public and private institutions / Regimes de flexibilización y sentidos del trabajo para docentes de enseñanza superior en instituciones públicas y privadas

Resumo Este artigo objetivou compreender o sentido do trabalho para o docente de ensino superior em instituições públicas e privadas em seus diversos regimes de flexibilização do trabalho, com base na identificação de elementos da racionalidade instrumental e subjetiva. Assim, a coleta de dados contou com fontes múltiplas, tais como a análise documental das instituições e dos contratos de trabalho dos docentes, a observação direta e as entrevistas realizadas presencialmente. Como resultado, percebeu-se que: há preponderância do sentido laboral substantivo sobre o sentido instrumental; a flexibilização altera a percepção da atuação individual e interfere no seu sentido substantivo; e a vida é arranjada a partir da organização do trabalho flexibilizado.

Abstract This article aims to understand the meaning of the work for the higher education teacher in public and private institutions in their various regimes of work flexibility based on the identification of instrumental and subjective rationality elements. The collection of data had multiple sources, such as the documentary analysis of institutions and teachers' work contracts, direct observation and personal interviews. As a result it was noticed that: there is a preponderance of the substantive meaning of the work on the instrumental sense; the flexibilization alters the perception of individual performance and infers on its substantive sense, and life is arranged from the organization of work flexibilized.

Resumen Este artículo objetivó comprender el sentido del trabajo para el docente de enseñanza superior en instituciones públicas y privadas en sus diversos regímenes de flexibilización del trabajo, a partir de la identificación de elementos de la racionalidad instrumental y subjetiva. Así, la recolección de datos contó con fuentes múltiples, tales como el análisis documental de las instituciones y de los contratos de trabajo de los docentes, la observación directa y entrevistas realizadas presencialmente. Como resultado se percibió que: hay preponderancia del sentido sustantivo del trabajo sobre el sentido instrumental; la flexibilización altera la percepción de la actuación individual e tiene impacto sobre su sentido sustantivo; y la vida se organiza a partir de la organización del trabajo flexibilizado.

LyeTxI-b, a Synthetic Peptide Derived From Lycosa erythrognatha Spider Venom, Shows Potent Antibiotic Activity in Vitro and in Vivo.

The antimicrobial peptide LyeTxI isolated from the venom of the spider Lycosa erythrognatha is a potential model to develop new antibiotics against bacteria and fungi. In this work, we studied a peptide derived from LyeTxI, named LyeTxI-b, and characterized its structural profile and its in vitro and in vivo antimicrobial activities. Compared to LyeTxI, LyeTxI-b has an acetylated N-terminal and a deletion of a His residue, as structural modifications. The secondary structure of LyeTxI-b is a well-defined helical segment, from the second amino acid to the amidated C-terminal, with no clear partition between hydrophobic and hydrophilic faces. Moreover, LyeTxI-b shows a potent antimicrobial activity against Gram-positive and Gram-negative planktonic bacteria, being 10-fold more active than the native peptide against Escherichia coli. LyeTxI-b was also active in an in vivo model of septic arthritis, reducing the number of bacteria load, the migration of immune cells, the level of IL-1ß cytokine and CXCL1 chemokine, as well as preventing cartilage damage. Our results show that LyeTxI-b is a potential therapeutic model for the development of new antibiotics against Gram-positive and Gram-negative bacteria.

Neutrophils: Beneficial and Harmful Cells in Septic Arthritis.

Septic arthritis is an inflammatory joint disease that is induced by pathogens such as Staphylococcus aureus. Infection of the joint triggers an acute inflammatory response directed by inflammatory mediators including microbial danger signals and cytokines and is accompanied by an influx of leukocytes. The recruitment of these inflammatory cells depends on gradients of chemoattractants including formylated peptides from the infectious agent or dying cells, host-derived leukotrienes, complement proteins and chemokines. Neutrophils are of major importance and play a dual role in the pathogenesis of septic arthritis. On the one hand, these leukocytes are indispensable in the first-line defense to kill invading pathogens in the early stage of disease. However, on the other hand, neutrophils act as mediators of tissue destruction. Since the elimination of inflammatory neutrophils from the site of inflammation is a prerequisite for resolution of the acute inflammatory response, the prolonged stay of these leukocytes at the inflammatory site can lead to irreversible damage to the infected joint, which is known as an important complication in septic arthritis patients. Thus, timely reduction of the recruitment of inflammatory neutrophils to infected joints may be an efficient therapy to reduce tissue damage in septic arthritis.

In Vitro TNF-α Inhibitory Activity of Brazilian Plants and Anti-Inflammatory Effect of Stryphnodendron adstringens in an Acute Arthritis Model.

Stryphnodendron species, popularly named "barbatimão," are traditionally used in Brazil as anti-inflammatory agents. This study aimed to investigate the effect of barbatimão and 11 other species on the production of tumor necrosis factor-alpha (TNF-α) in lipopolysaccharide- (LPS-) stimulated THP-1 cells, as well as their anti-arthritis activity. The extracts of Stryphnodendron adstringens, Stryphnodendron obovatum, Campomanesia lineatifolia, and Terminalia glabrescens promoted a concentration-dependent inhibition of TNF-α. Mice injected with LPS in the knee joint were treated per os with fractions from the selected extracts. Both the organic (SAO) and the aqueous (SAA) fractions of S. adstringens promoted a dose-dependent reduction of leukocyte migration and neutrophil accumulation into the joint, but none of them reduced CXCL1 concentration in the periarticular tissue. In contrast, treatment with C. lineatifolia and T. glabrescens fractions did not ameliorate the inflammatory parameters. Analyses of SAO by Ultra Performance Liquid Chromatography (UPLC) coupled to electrospray ionization mass spectrometry (ESI-MS) led to the identification of gallic acid along with 11 prodelphinidins, characterized as monomers and dimers of the B-type. Our findings contribute to some extent to corroborating the traditional use of S. adstringens as an anti-inflammatory agent. This activity is probably related to a decrease of leukocyte migration into the inflammatory site. Polyphenols like gallic acid and prodelphinidins, identified in the active fraction, may contribute to the observed activity.

Effect of the Hydroethanolic Extract from Echinodorus grandiflorus Leaves and a Fraction Enriched in Flavone-C-Glycosides on Antigen-Induced Arthritis in Mice.

The leaves of Echinodorus grandiflorus are traditionally used in Brazil to treat several inflammatory conditions, including arthritis. This study aimed to investigate the antiarthritis activity of the 70% ethanol extract of E. grandiflorus leaves and a standardized flavonoid-rich fraction in an antigen-induced arthritis model in mice. Previously immunized mice were treated per os with saline (control group), 70% ethanol extract (100-1000 mg/kg), or a flavonoid-rich fraction (0.7-7.2 mg/kg) 40 minutes before and 3 and 6 hours after the challenge with antigen into the knee joint. The administration of the 70% ethanol extract and flavonoid-rich fraction to mice significantly reduced neutrophil recruitment to the joint cavity and in periarticular tissue. The levels of chemokine (C-X-C motif) ligand 1, tumor necrosis factor-α, and interleukin-1ß quantified by the enzyme-linked immunosorbent assay (ELISA) in the periarticular tissue were also diminished in mice treated with the 70% ethanol extract and flavonoid-rich fraction, as well as mechanical hypernociception. Histological analysis confirmed that both the 70% ethanol extract and flavonoid-rich fraction suppressed joint inflammation and inhibited cartilage and bone destruction when compared to the control group. Our results demonstrate, for the first time, that E. grandiflorus has anti-inflammatory activity in an experimental arthritis model and highlights the role of flavonoids in the observed response.

Activities of 2-phthalimidethanol and 2-phthalimidethyl nitrate, phthalimide analogs devoid of the glutarimide moiety, in experimental models of inflammatory pain and edema.

The reintroduction of thalidomide in the pharmacotherapy greatly stimulated the interest in the synthesis and pharmacological evaluation of phthalimide analogs with new and improved activities and also greater safety. In the present study, we evaluated the activities of two phthalimide analogs devoid of the glutarimide ring, namely 2-phthalimidethanol (PTD-OH) and 2-phthalimidethyl nitrate (PTD-NO), in experimental models of inflammatory pain and edema in male C57BL/6J mice. Intraplantar (i.pl.) injection of carrageenan (300 µg) induced mechanical allodynia and this response was inhibited by previous per os (p.o.) administration of PTD-OH and PTD-NO (750 mg/kg) and also by thalidomide (500 or 750 mg/kg). The edema induced by carrageenan was also inhibited by previous p.o. administration of PTD-OH (500 and 750 mg/kg) and PTD-NO (125, 250, 500 or 750 mg/kg), but not by thalidomide. Carrageenan increased tumor necrosis factor (TNF)-α and CXCL1 concentrations and also the number of neutrophils in the paw tissue. Previous p.o. administration of PTD-NO (500 mg/kg) reduced all the parameters, while PTD-OH (500 mg/kg) reduced only the accumulation of neutrophils. Thalidomide, on the other hand, was devoid of effect on these biochemical parameters. Plasma concentrations of nitrite were increased after p.o. administration of the phthalimide analog coupled to a NO donor, PTD-NO (500 mg/kg), but not after administration of PTD-OH or thalidomide. In conclusion, our results show that small molecules, structurally much simpler than thalidomide or many of its analogs under investigation, exhibit similar activities in experimental models of pain and inflammation. Finally, as there is evidence that the glutarimide moiety contributes to the teratogenic effect of many thalidomide analogs, our results indicate that phthalimide analogs devoid of this functional group could represent a new class of analgesic and anti-inflammatory candidates with potential greater safety.

Perfil dos candidatos inaptos para doação de sangue no serviço de hemoterapia do Hospital Santo Ângelo, RS, Brasil / Profile of ineligible candidates for blood donation in the hemotherapy service of Santo Ângelo Hospital, RS, Brazil

O avanço das metodologias de diagnóstico das doenças transmissíveis pelo sangue e o desenvolvimento da triagem clínica na seleção dos candidatos à doação de sangue tornaram a prática hemoterápica mais segura. As campanhas de doação de sangue buscam atrair maior número decandidatos para atender à demanda de produtos hemoterápicos. Para isso, no entanto, o candidato deve passar pela triagem clínica que inclui uma entrevista socioepidemiológica a fim de selecionaraquele que for saudável, de modo que seja produzido o melhor produto hemoterápico possível. Por meio da análise de 505 fichas de candidatos considerados inaptos pela triagem clínica entre 2005 e 2010, no Serviço de Hemoterapia de Santo Ângelo, Rio Grande do Sul, buscou-se identificar o perfil do candidato inapto à doação. Deste total, 62por cento eram do gênero masculino e, em ambos os gêneros,o maior número de inaptos tinha ensino médio completo e idade entre 18 e 23 anos. As principais causas de inaptidão foram a multiplicidade de parceiros sexuais e o comportamento de risco para o sexo masculino e hematócrito baixo para o feminino. Conhecer o perfil do doador inapto chama a atenção para a importância das informações colhidas durante a triagem clínica, torna o entrevistador mais preparado e a prática hemoterápica mais segura.

Prevalência de VDRL reagente em doadores do hemocentro regional de Cruz Alta – RS, Brasil, no período de 2003 a 2009 / Prevalence of positive VDRL in donors of the blood center in Cruz Alta municipality, Brazil, from 2003 to 2009

A sífilis é uma doença causada pela bactéria Treponema pallidum e clinicamente dividida em primária, secundária e terciária. A transmissão se dá principalmente pelas vias sexual e congênita e por transfusões sanguíneas. A sorologia para sífilis em doadores de sangue é realizada desde 1938 e o objetivo deste estudo foi verificar a prevalência do teste Veneral Disease Research Laboratory (VDRL) reagente em doadores do Hemocentro Regional de Cruz Alta no Rio Grande do Sul, no período de agosto de 2003 a julho de 2009. Foram analisadas 20.780 doações e consideradas, neste estudo, como as que apresentaram sorologia reagente e inconclusiva para o teste VDRL. A prevalência encontrada foi de 0,3por cento para sorologia reagente e 0,13por cento para inconclusiva. Em relação ao gênero dos doadores reagentes, 58,33por cento foram do sexo masculino. A faixa etária com maior reatividade foi de 31 e 40 anos. Conclui-se que a prevalência de 0,3por cento de VDRL reagente encontrada neste estudo é considerada baixa e que vem diminuindo nos últimos anos. Os resultados deste estudo podem ser utilizados para o desenvolvimento de ações públicas para prevenção e controle da prevalência da sífilis na região.