Amazonian Melastomataceae blueberries: Determination of phenolic content, nutritional composition, and antioxidant and anti-glycation activities.

Berries come from hundreds of different species of plants spread around the world. Blackberries, blueberries and raspberries, for instance, are popular berries that have attracted attention for providing several benefits to human health. Wild berries from the Melastomataceae family are commonly encountered in the Amazon, although these small blue fruits are poorly consumed. Although domesticated fruits give better monetary profits, the consumption of wild fruits is a desirable option to afford income and/or food to communities at the same time as keep the Amazon region preserved. Aiming the divulgation of the nutritional potential of these plants, this paper describes the study of six species of Amazonian blueberries, five of them from the Clidemia genus and one from the Tococa genus, in regard to their nutritional and chemical composition and antioxidant activity (AA). The levels of moisture, ash, protein, lipids, carbohydrates, and the total caloric values obtained for the Amazonian blueberries were comparable to other common edible berries. Although the six species are similar in terms of nutritional composition, their anthocyanin profiles and contents are quite peculiar. Two non-methylated anthocyanins, cyanidin and delphinidin, which bound to a variable number of sugars, characterized the berries of the genera Clidemia and Tococa. Clidemia japurensis, Clidemia hirta and Tococa bullifera were rich in tri-glycosylated anthocyanins, although differences are notable between them. Clidemia pustulata and Clidemia capitellata were characterized by the prevalence of mono-glycosylated anthocyanins, and Clidemia rubra showed a unique profile with mono- and di-glycosylated homologous as the main anthocyanins. In addition to their different chemical profiles, the concentrations of anthocyanins and other phenolic compounds varied a lot among the six species studied. The species C. rubra had the highest total concentration of phenolic acids and flavonoids. Therefore, this study showed that the blueberries analyzed have potential to be better explored, which we suggest doing in a sustainable way, aiming at the preservation of the Amazon's biodiversity.

Recent Advances in Host-Directed Therapies for Tuberculosis and Malaria.

Tuberculosis (TB), caused by the bacterium Mycobacterium tuberculosis, and malaria, caused by parasites from the Plasmodium genus, are two of the major causes of death due to infectious diseases in the world. Both diseases are treatable with drugs that have microbicidal properties against each of the etiologic agents. However, problems related to treatment compliance by patients and emergence of drug resistant microorganisms have been a major problem for combating TB and malaria. This factor is further complicated by the absence of highly effective vaccines that can prevent the infection with either M. tuberculosis or Plasmodium. However, certain host biological processes have been found to play a role in the promotion of infection or in the pathogenesis of each disease. These processes can be targeted by host-directed therapies (HDTs), which can be administered in conjunction with the standard drug treatments for each pathogen, aiming to accelerate their elimination or to minimize detrimental side effects resulting from exacerbated inflammation. In this review we discuss potential new targets for the development of HDTs revealed by recent advances in the knowledge of host-pathogen interaction biology, and present an overview of strategies that have been tested in vivo, either in experimental models or in patients.

Mycobacterium tuberculosis Induces Irg1 in Murine Macrophages by a Pathway Involving Both TLR-2 and STING/IFNAR Signaling and Requiring Bacterial Phagocytosis.

Irg1 is an enzyme that generates itaconate, a metabolite that plays a key role in the regulation of inflammatory responses. Previous studies have implicated Irg1 as an important mediator in preventing excessive inflammation and tissue damage in Mycobacterium tuberculosis (Mtb) infection. Here, we investigated the pattern recognition receptors and signaling pathways by which Mtb triggers Irg1 gene expression by comparing the responses of control and genetically deficient BMDMs. Using this approach, we demonstrated partial roles for TLR-2 (but not TLR-4 or -9), MyD88 and NFκB signaling in Irg1 induction by Mtb bacilli. In addition, drug inhibition studies revealed major requirements for phagocytosis and endosomal acidification in Irg1 expression triggered by Mtb but not LPS or PAM3CSK4. Importantly, the Mtb-induced Irg1 response was highly dependent on the presence of the bacterial ESX-1 secretion system, as well as host STING and Type I IFN receptor (IFNAR) signaling with Type II IFN (IFN-γ) signaling playing only a minimal role. Based on these findings we hypothesize that Mtb induces Irg1 expression in macrophages via the combination of two independent triggers both dependent on bacterial phagocytosis: 1) a major signal stimulated by phagocytized Mtb products released by an ESX-1-dependent mechanism into the cytosol where they activate the STING pathway leading to Type I-IFN production, and 2) a secondary TLR-2, MyD88 and NFκB dependent signal that enhances Irg1 production independently of Type I IFN induction.

Investigando, prevenindo e tratando a Cegueira Botânica em diferentes cenários do Estado do Rio de Janeiro / Investigating, preventing and treating Botanical Blindness in different scenarios in the State of Rio de Janeiro

As plantas são importantes para a biosfera e para os seres humanos de diversas maneiras. Entretanto, passam despercebidas para a maioria das pessoas, o que é conhecido como "Cegueira Botânica" (CB), que ameaça a existência de todos seres vivos. O Ensino de Botânica (EB), que poderia auxiliar a reverter esse quadro, é na atualidade, segundo vários autores, conteudista, descontextualizado, desatualizado e marginal embora faça parte dos currículos do ensino médio, no Brasil. Nesse contexto, foi realizada a presente pesquisa que tem como objetivo geral investigar a existência da CB em diversos cenários do Município do Rio de Janeiro e sua relação com o EB. Como metodologia foi realizada uma pesquisação sobre a CB e EB em vários documentos oficiais, no nível médio em duas escolas, nos livros didáticos e em agumas Unidades de Conservação do Município do Rio de Janeiro. Constituíram o público participante dessa pesquisa, 98 alunos e 24 professores do 3º ano, de 10 turmas, 16 botânicos e 60 escaladores do Estado do Rio de Janeiro. Os instrumentos de coleta de dados foram questionários com perguntas abertas e fechadas, observação direta de aulas práticas de laboratório, de campo e atividades de extensão. Os resultados indicaram que embora o conteúdo e a prática do EB nas turmas de nível médio das escolas analisadas sejam diferentes do ponto de vista pedagógico e sejam de excelência, a CB existe e acomete a maioria dos alunos, dos professores em diferentes graus, apesar da boa relação que a maioria tem com as plantas e com o seu ensino. A maioria dos escaladores notam as plantas, ou seja, não sofrerem de cegueira botânica, mas não sabem reconhecer as espécies. Os resultados permitiram identificar também que a relação da maioria dos botânicos analisados têm com as plantas serve de modelo para a sociedade. Nesse contexto, foi possível concluir que, apesar da existência da CB em diferentes graus nos públicos participantes da pesquisa, eles tem uma boa relação com as plantas e com o EB, mas tanto o ensino como a relação podem melhorar e a CB deve ser tratada e prevenida. Materiais e atividades para o tratamento e prevenção para a CB foram propostos.

Plants are important to the biosphere and to humans in many ways. However, they go unnoticed for most people, what is known as "Plant Blindness" (PB), that threatens the existence of all living beings. The Teaching of Botany (TB), which could help to reverse this situation, is currently, according to several authors, contentist, decontextualized, outdated and marginal although it is part of the high school curricula in Brazil. In this context, this research aimed to investigate the existence of PB in various scenarios of the city of Rio de Janeiro and its relationship with TB. For this, as a methodology, a qualitative research on PB and TB was carried out in several official documents, at the middle level in two schools, in the textbooks and in other Conservation Units of the Municipality of Rio de Janeiro. The participants of this research were 98 students and 24 3rd grade teachers from 10 groups, 16 botanists and 60 climbers from the State of Rio de Janeiro. The data collection instruments were questionnaires with open and closed questions, direct observation of laboratory, field and extension activities. The results indicated that although the content and practice of EB in the middle classes of the schools analyzed are different from the pedagogical point of view and are of excellence, the CB exists and affects the majority of students, teachers in different degrees, despite the good relationship that most have with plants and their teaching. Most climbers notice the plants, that is, they do not suffer from botanical blindness, but they do not know how to recognize the species. The results also made it possible to identify that the relationship most of the analyzed botanists have with plants serves as a model for society. In this context, it was possible to conclude that, despite the existence of CB in different degrees in the public participating in the research, they have a good relationship with plants and with EB, but both teaching and the relationship can improve and CB must be treated and prevented. Materials and activities for the treatment and prevention of BC have been proposed.

The mycma_1113 Gene from Mycobacterium abscessus subsp. massiliense is Related to Siderophore Synthesis.

Iron (Fe) homeostasis control is important for both pathogen and the host. During infection, the host reduces the access of microorganisms to iron, however, studies have shown that virulent pathogens are capable to sequester Fe from host proteins, and establish the infection. M. abscessus subsp. massiliense (Mycma), that is resistant to most drugs used against tuberculosis, was responsible for outbreaks around the world showing increased virulence when compared to other rapidly growing mycobacteria. The goal of this study was to determine whether Mycma produce siderophores and if the mycma_1113 gene expression, a putative homolog of M. tuberculosis mbtB gene located in the mbt gene cluster, is related to the synthesis of these molecules. For that, the effect of different iron concentrations on the growth of Mycma, the expression of mycma_1113 gene, and the production of siderophores was evaluated in vitro and in vivo. It is shown that Mycma produce siderophores under iron deprivation conditions and mycma_1113 gene expression was influenced by iron availability. The mycma_1113 gene expression was also increased after macrophage or in vivo infection indicating that mycobactin synthesis by Mycma could participate in the Fe sequestration from the host during infection. In conclusion, we show that Mycma produces siderophores under iron deprivation conditions and that the mycma_1113 gene is involved in this process, furthermore, this gene expression is induced during infection.

The Activity of a Hexameric M17 Metallo-Aminopeptidase Is Associated With Survival of Mycobacterium tuberculosis.

Mycobacterium tuberculosis is one of the most prevalent human pathogens causing millions of deaths in the last years. Moreover, tuberculosis (TB) treatment has become increasingly challenging owing to the emergence of multidrug resistant M. tuberculosis strains. Thus, there is an immediate need for the development of new anti-TB drugs. Proteases appear to be a promising approach and may lead to shortened and effective treatments for drug-resistant TB. Although the M. tuberculosis genome predicts more than 100 genes encoding proteases, only a few of them have been studied. Aminopeptidases constitute a set of proteases that selectively remove amino acids from the N-terminus of proteins and peptides and may act as virulence factors, essential for survival and maintenance of many microbial pathogens. Here, we characterized a leucine aminopeptidase of M. tuberculosis (MtLAP) as a cytosolic oligomeric metallo-aminopeptidase. Molecular and enzymatic properties lead us to classify MtLAP as a typical member of the peptidase family M17. Furthermore, the aminopeptidase inhibitor bestatin strongly inhibited MtLAP activity, in vitro M. tuberculosis growth and macrophage infection. In murine model of TB, bestatin treatment reduced bacterial burden and lesion in the lungs of infected mice. Thus, our data suggest that MtLAP participates in important metabolic pathways of M. tuberculosis necessary for its survival and virulence and consequently may be a promising target for new anti-TB drugs.

Mycobacterium tuberculosis Prolyl Oligopeptidase Induces In vitro Secretion of Proinflammatory Cytokines by Peritoneal Macrophages.

Tuberculosis (TB) is a disease that leads to death over 1 million people per year worldwide and the biological mediators of this pathology are poorly established, preventing the implementation of effective therapies to improve outcomes in TB. Host-bacterium interaction is a key step to TB establishment and the proteases produced by these microorganisms seem to facilitate bacteria invasion, migration and host immune response evasion. We presented, for the first time, the identification, biochemical characterization, molecular dynamics (MDs) and immunomodulatory properties of a prolyl oligopeptidase (POP) from Mycobacterium tuberculosis (POPMt). POP is a serine protease that hydrolyzes substrates with high specificity for proline residues and has already been characterized as virulence factor in infectious diseases. POPMt reveals catalytic activity upon N-Suc-Gly-Pro-Leu-Gly-Pro-AMC, a recognized POP substrate, with optimal activity at pH 7.5 and 37°C. The enzyme presents KM and Kcat/KM values of 108 µM and 21.838 mM-1 s-1, respectively. MDs showed that POPMt structure is similar to that of others POPs, which consists of a cylindrical architecture divided into an α/ß hydrolase catalytic domain and a ß-propeller domain. Finally, POPMt was capable of triggering in vitro secretion of proinflammatory cytokines by peritoneal macrophages, an event dependent on POPMt intact structure. Our data suggests that POPMt may contribute to an inflammatory response during M. tuberculosis infection.

Heart Rate Variability Analysis in an Experimental Model of Hemorrhagic Shock and Resuscitation in Pigs.

BACKGROUND: The analysis of heart rate variability (HRV) has been shown as a promising non-invasive technique for assessing the cardiac autonomic modulation in trauma. The aim of this study was to evaluate HRV during hemorrhagic shock and fluid resuscitation, comparing to traditional hemodynamic and metabolic parameters. METHODS: Twenty anesthetized and mechanically ventilated pigs were submitted to hemorrhagic shock (60% of estimated blood volume) and evaluated for 60 minutes without fluid replacement. Surviving animals were treated with Ringer solution and evaluated for an additional period of 180 minutes. HRV metrics (time and frequency domain) as well as hemodynamic and metabolic parameters were evaluated in survivors and non-survivors animals. RESULTS: Seven of the 20 animals died during hemorrhage and initial fluid resuscitation. All animals presented an increase in time-domain HRV measures during haemorrhage and fluid resuscitation restored baseline values. Although not significantly, normalized low-frequency and LF/HF ratio decreased during early stages of haemorrhage, recovering baseline values later during hemorrhagic shock, and increased after fluid resuscitation. Non-surviving animals presented significantly lower mean arterial pressure (43±7 vs 57±9 mmHg, P<0.05) and cardiac index (1.7±0.2 vs 2.6±0.5 L/min/m2, P<0.05), and higher levels of plasma lactate (7.2±2.4 vs 3.7±1.4 mmol/L, P<0.05), base excess (-6.8±3.3 vs -2.3±2.8 mmol/L, P<0.05) and potassium (5.3±0.6 vs 4.2±0.3 mmol/L, P<0.05) at 30 minutes after hemorrhagic shock compared with surviving animals. CONCLUSIONS: The HRV increased early during hemorrhage but none of the evaluated HRV metrics was able to discriminate survivors from non-survivors during hemorrhagic shock. Moreover, metabolic and hemodynamic variables were more reliable to reflect hemorrhagic shock severity than HRV metrics.

The endothelin system has a significant role in the pathogenesis and progression of Mycobacterium tuberculosis infection.

Tuberculosis (TB) remains a major global health problem, and although multiple studies have addressed the relationship between Mycobacterium tuberculosis and the host on an immunological level, few studies have addressed the impact of host physiological responses. Proteases produced by bacteria have been associated with important alterations in the host tissues, and a limited number of these enzymes have been characterized in mycobacterial species. M. tuberculosis produces a protease called Zmp1, which appears to be associated with virulence and has a putative action as an endothelin-converting enzyme. Endothelins are a family of vasoactive peptides, of which 3 distinct isoforms exist, and endothelin 1 (ET-1) is the most abundant and the best-characterized isoform. The aim of this work was to characterize the Zmp1 protease and evaluate its role in pathogenicity. Here, we have shown that M. tuberculosis produces and secretes an enzyme with ET-1 cleavage activity. These data demonstrate a possible role of Zmp1 for mycobacterium-host interactions and highlights its potential as a drug target. Moreover, the results suggest that endothelin pathways have a role in the pathogenesis of M. tuberculosis infections, and ETA or ETB receptor signaling can modulate the host response to the infection. We hypothesize that a balance between Zmp1 control of ET-1 levels and ETA/ETB signaling can allow M. tuberculosis adaptation and survival in the lung tissues.

Efeitos do metoprolol em modelo experimental de choque séptico / Effects of metoprolol in experimental model of septic shock

O uso de fármacos cardiovasculares tem aumentado consideravelmente nos últimos anos, incluindo o uso de beta-bloqueadores como o metoprolol. E embora estudos experimentais e clínicos tenham demonstrado benefícios na utilização desta classe de fármacos em pacientes sépticos, o bloqueio de receptores beta-adrenérgicos continua sendo contraditório, especialmente no choque séptico. Vinte fêmeas suínas nas quais o choque séptico foi induzido através da infusão intravenosa de Escherichia coli (6x109 UFC/kg em duas horas), foram randomicamente distribuídas (n=10 por grupo) em um grupo Metoprolol (GM; 214.2 ?g/kg de metoprol infundido em 45 minutos) ou grupo Controle (GC), o qual recebeu um volume correspondente de solução salina. Os parâmetros hemodinâmicos e de oxigenação foram avaliados no momento basal (TB), T30, T60, T120, T240 e T360 minutos. Amostras de sangue foram colhidas ainda em TB, T120 e T360 e armazenadas para posterior mensuração da concentração sérica de citocinas e marcadores cardíacos. Durante este período utilizou-se um protocolo de ressuscitação com Ringer lactato, norepinefrina (NE) e dobutamina para manutenção da pressão arterial média (PAM) > 65 mmHg, da pressão venosa central (PVC) em 8-12 mmHg e da saturação venosa mista de oxigênio (SvO2) > 65%. Os dados paramétricos foram analisados utilizando ANOVA de duas vias para medidas repetidas, seguidas por Tukey quando necessário, enquanto para os dados não paramétricos utilizaram-se os testes de Kruskall-Wallis e Mann-Whitney. A quantidade de fluido, NE e dobutamina foi analisada pelo teste t de Student, e a análise de sobrevivência foi realizada utilizando o Kaplan-Meier log-rank test. Exceto por um valor mais elevado do índice de resistência vascular sistêmica (IRVS) em T240 (p=0,02) nos animais tratados com metoprolol, todos os parâmetros analisados neste estudo apresentaram um comportamento similar em ambos os grupos. Nenhuma diferença foi observada também em relação ao volume total de fluido...

The use of cardiovascular drugs considerably increased in recent years, including the use of ?-blockers, such as metoprolol. And although experimental and clinical studies had demonstrated benefits on the administration of these drugs in septic patients, the ?-blockade is still contradictory, especially in the septic shock. Twenty female pigs in which septic shock was induced through intravenous E. coli infusion (6x109 c.f.u/kg in 2h) were randomly assigned (n=10 per group) to the Metoprolol group (214.2 ?g kg-1 of metoprol infused in 45 minutes) or Control group, which received a correspondent volume of normal saline. Hemodynamic and oxygenation parameters were then evaluated at baseline (TB), T30, T60, T120, T240 and T360 minutes. Blood samples were collected at TB, T120 and T360 for posterior mensuration of serum cytokines and cardiac markers. During this period, a resuscitation protocol with lactated Ringer's solution, norepinephrine and dobutamine was used to maintain the mean arterial pressure (MAP) > 65 mmHg, the central venous pressure (CVP) between 8-12 mmHg and the mixed venous oxygen saturation (SvO2) > 65%. Parametric data were compared using the two-way repeated measures ANOVA, followed by the Tukey test when necessary, while the nonparametric data were analyzed with the Kruskall-Wallis and the Mann-Whitney test. The total volume of fluid and total amount of norepinephrine and dobutamine infused were analyzed by the Student's t-test. Survival analysis was performed using Kaplan-Meier log-rank test. Except for the higher values of systemic vascular resistance index (SVRI) at T240 (p=0.02) in the animals that received metoprolol, all the parameters analyzed in this study showed a similar behavior in both groups. No difference was also observed between groups in relation to the total volume of fluid (p=0.914), the total amount of norepinephrine (p=0.069) and dobutamine (p=0.560) infused, and related to the survival rate. Although...

Respostas cardiorrespiratória e metabólica do propofol nas formulações em emulsão lipídica ou microemulsão em gatas / Cardiorespiratory and metabolic answer with microemulsion and lipid emulsion of propofol in cats

O presente estudo objetiva avaliar os efeitos cardiorrespiratórios e metabólicos do propofol em emulsão lipídica e microemulsão em gatas. Foram utilizadas 12 gatas, hígidas, adultas, alocadas em dois grupos: microemulsão (MICRO, n=6) e emulsão lipídica (EMU, n=6), os quais receberam propofol, na respectiva formulação, em dose suficiente para intubação. Em seguida, foram intubados, fornecendo-se oxigênio 100%, em sistema sem reinalação de gases. Ato contínuo, iniciou-se a infusão de propofol na dose de 0,3mg kg-1 min-1 durante 90 minutos. A dose necessária para indução foi de 9,5±1,3mg kg-1 e 10±1mg kg-1 para MICRO e EMU, respectivamente. Os valores de pressão arterial sistólica (PAS), pressão arterial média (PAM), pressão arterial diastólica (PAD) e pH foram menores, em todos os momentos, no EMU em relação ao MICRO; a f do EMU foi menor de T30 até T75 em relação ao MICRO. A PaCO2 do EMU foi maior de T15 até T90. Os tempos de extubação, decúbito esternal, deambulação e recuperação total foram de 40,6±30,7; 91±37,5; 134,5±54,5 e 169,1±55,4 minutos no MICRO e de 68,8±37,3; 133,3±85,3; 171,3±77,1 e 233,1±60,6 minutos no EMU, respectivamente. Houve aumento da enzima Alanina Aminotransferase de 12 às 72h no EMU e de 48 às 72h no MICRO. O propofol em microemulsão apresentou características clínicas de indução e manutenção, bem como efeitos metabólicos semelhantes à formulação em emulsão lipídica. A formulação em microemulsão proporcionou maior estabilidade cardiovascular e respiratória para indução e infusão contínua em gatas hígidas.

The aim of the present study was to evaluate the cardiorespiratory and metabolic effects from lipid emulsion and microemulsion of propofol in cats. Twelve healthy adult cats were included, and divided into two groups: microemulsion group (MICRO, n=6) and lipid emulsion (EMU, n=6), where they received propofol in the respective formulation, in a dose sufficient for intubation. The animals were then intubated and provided with 100% oxygen through a non-rebreathing circuit. Immediately after, the infusion of propofol was initiated (0.3mg kg-1 min-1) and maintained for 90 minutes. The dose required for induction was 9.5±1.3mg kg-1 and 10±1mg kg-1 in MICRO and EMU, respectively. The SAP, MAP, DAP and pH values were lower in all moments in EMU when compared to MICRO; the RR in EMU was lower from T30 to T75 in comparison to MICRO. The paCO2 was greater in the EMU from T15 to T90. The times to extubation, sternal recumbency, ambulation and total recovery were 40.6±30.7, 91±37.5, 134.5±54.5 and 169.1±55.4 minutes in MICRO and 68.8±37.3, 133.3±85.3, 171.3±77.1 and 233.1±60.6 minutes in EMU, respectively. There was an increase of the enzyme alanine aminotransferase from 12 to 72 hours in EMU and in MICRO from 48 to 72 hours. The propofol in microemulsion presents clinical characteristics of induction and maintenance, and metabolic effects similar to the formulation in lipid emulsion. The microemulsion formulation provides a better cardiovascular and respiratory stability for induction and continuous infusion in healthy cats.

Evaluation of the clinical and cardiorespiratory effects of propofol microemulsion in dogs / Efeitos clínicos e cardiorespiratórios do propofol em microemulsão em cães

This research aimed to evaluate the clinical and cardiorespiratory effects of a propofol formulation with nanometer droplet diameter in dogs. Six adult healthy female dogs weighing 14.8±1.2kg were used in this study. Each dog received two treatments with a 15-day washout period. A microemulsion (MICRO) or lipid emulsion (EMU) of propofol was administered intravenously (IV) for induction and maintenance of anesthesia. Anesthesia was maintained with a constant rate infusion of propofol (0.4mg kg-1 minute-1). Cardiorespiratory variables were recorded before induction (baseline), immediately after and at 15-minute intervals for 90 minutes after treatment. Arterial blood samples were also taken for blood gas analysis, except at 45 and 75 minutes after induction. The mean arterial pressure decreased significantly during both treatments, while the cardiac index decreased significantly only in MICRO treatment. The time to extubation, sternal recumbency, ambulation and total recovery was similar in both treatments. Opisthotonos was observed in 33% of the animals in each treatment. The propofol microemulsion presented clinical and respiratory parameters similar to those obtained with the lipid emulsion commercially available, but had some significantly different hemodynamic characteristics when used for inducing and maintaining anesthesia. Based only in these results, no advantages are seen in the use of this new microemulsion.

Este estudo tem o objetivo de avaliar os efeitos clínicos e cardiorrespiratórios de uma formulação de propofol formada por nanopartículas, em cães. Para esse propósito, seis cães hígidos, adultos, fêmeas, com peso médio de 14,8±1,2kg foram utilizados. Cada cão recebeu dois tratamentos, sendo que entre estes foi permitido aos animais um período de washout de 15 dias. Os animais receberam propofol em microemulsão (MICRO) ou em emulsão lipídica (EMU) por via intravenosa (IV) para a indução e manutenção da anestesia. A anestesia foi mantida com velocidade de infusão constante (0,4mg kg-1 min-1). As variáveis cardiorrespiratórias foram mensuradas antes da indução (basal), imediatamente após a indução, e então a cada 15 minutos, durante 90 minutos. Amostras de sangue arterial também foram colhidas para análise de gases sanguíneos, exceto aos 45 e 75 minutos após a indução. A pressão arterial média diminuiu significativamente com a utilização de ambos os tratamentos, enquanto o índice cardíaco reduziu significativamente somente com o tratamento MICRO. O tempo necessário para extubação, decúbito esternal, deambulação e recuperação total foi semelhante em ambos os grupos. Opistótono foi observado em 33% dos animais em cada tratamento. A microemulsão de propofol apresentou parâmetros clínicos e respiratórios semelhantes àqueles obtidos com a emulsão lipídica comercialmente disponível, porém mostrou algumas características hemodinâmicas diferentes, quando utilizada para a indução e manutenção da anestesia. Com base somente nesses resultados, não são observadas vantagens na utilização da microemulsão.

Type C botulism in swine fed on restaurant waste / Botulismo tipo C em suínos alimentados com restos de restaurante

Descreve-se a epidemiologia da morte de suínos ocorrida entre 2002 e 2009 pela ingestão da neurotoxina botulínica do tipo C. A neurotoxina presente em resíduos de alimentos provenientes de cozinhas de restaurantes e hotéis, armazenados em tonéis sem abrigo do sol e administrado em cocho coletivo sem tratamento térmico prévio. Animais de diferentes idades morreram com sinais clínicos de botulismo caracterizados por paralisia flácida, emagrecimento, anorexia, fraqueza, incoordenação, dificuldade de locomoção com evolução para decúbito lateral com micção e defecação involuntárias. Nenhuma alteração evidente foi observada na necropsia e no exame histológico. O teste de bioensaio com soroneutralização em camundongos foi realizado com amostras de conteúdo intestinal dos animais afetados e alimentos e revelou a presença de grandes quantidades de toxina botulínica do tipo C.

The paper addresses the epidemiologic data of the death of pigs during the period of 2002 to 2009 following the ingestion of botulinum neurotoxin type C. This neurotoxin was present in food residues originating from restaurant and hotel kitchens, stored in barrels without shelter from the sun and administered in a collective trough without prior thermal treatment. Animals which died at different ages showed clinical signs of botulism characterized by flaccid paralysis, weight loss, anorexia, weakness, lack of coordination, locomotion difficulties with the evolution of lateral recumbency with involuntary urination and defecation. No alterations were observed at postmortem and histological examination. The bioassay with serum neutralization in mice was carried out on samples of intestinal contents from pigs affected and revealed the presence of large quantities of botulinum toxin type C.

Infections with avian pathogenic and fecal Escherichia coli strains display similar lung histopathology and macrophage apoptosis.

The purpose of this study was to compare histopathological changes in the lungs of chickens infected with avian pathogenic (APEC) and avian fecal (A(fecal)) Escherichia coli strains, and to analyze how the interaction of the bacteria with avian macrophages relates to the outcome of the infection. Chickens were infected intratracheally with three APEC strains, MT78, IMT5155, and UEL17, and one non-pathogenic A(fecal) strain, IMT5104. The pathogenicity of the strains was assessed by isolating bacteria from lungs, kidneys, and spleens at 24 h post-infection (p.i.). Lungs were examined for histopathological changes at 12, 18, and 24 h p.i. Serial lung sections were stained with hematoxylin and eosin (HE), terminal deoxynucleotidyl dUTP nick end labeling (TUNEL) for detection of apoptotic cells, and an anti-O2 antibody for detection of MT78 and IMT5155. UEL17 and IMT5104 did not cause systemic infections and the extents of lung colonization were two orders of magnitude lower than for the septicemic strains MT78 and IMT5155, yet all four strains caused the same extent of inflammation in the lungs. The inflammation was localized; there were some congested areas next to unaffected areas. Only the inflamed regions became labeled with anti-O2 antibody. TUNEL labeling revealed the presence of apoptotic cells at 12 h p.i in the inflamed regions only, and before any necrotic foci could be seen. The TUNEL-positive cells were very likely dying heterophils, as evidenced by the purulent inflammation. Some of the dying cells observed in avian lungs in situ may also be macrophages, since all four avian E. coli induced caspase 3/7 activation in monolayers of HD11 avian macrophages. In summary, both pathogenic and non-pathogenic fecal strains of avian E. coli produce focal infections in the avian lung, and these are accompanied by inflammation and cell death in the infected areas.

Immunohistochemical and ultra-structural detection of Pneumocystis in wild boars (Sus scrofa) co-infected with porcine circovirus type 2 (PCV2) in Southern Brazil.

Pneumocystis spp. are fungi that are able to infect a variety of host species and, occasionally, lead to severe pneumonia. Porcine circovirus type 2 (PCV2) is an important viral pathogen which affects both swine and wild boar herds worldwide. Co-infection between PCV2 and other pathogens has been reported, and the secondary immunodeficiency caused by the virus may predispose to these co-infections. In the present study, postmortem tissue samples obtained from wild boar herds in Southern Brazil were analyzed by histopathology, ultra-structural observation, and immunohistochemistry. Forty-seven out of seventy-eight (60%) wild boars showed clinical signs, gross, and histopathological lesions characteristic of infection by PCV2. Pneumocystis was detected by immunohistochemistry in 39 (50%) lungs and viral antigens of PCV2 were found in 29 (37.2%) samples. Concomitant presence of Pneumocystis and PCV2 were observed in 16 (20.5%) of the wild boars. Cystic and trophic forms of Pneumocystis were similar to previously described ultra-structural observations in other mammals.

Long-term cyclosporine treatment: evaluation of serum biochemical parameters and histopathological alterations in Wistar rats.

The immunosuppressant agent cyclosporine (CsA) is currently used in transplanted patients and in the therapy of autoimmune disorders. CsA treatment has significant acute and chronic side effects on the liver and kidney. However, in the clinical setting, it is difficult to distinguish a direct effect of CsA treatment from other confounding variables, such as allograft rejection and effects due to other drug therapies. In the present study, we assessed for direct associations between CsA immunosuppressive therapy and cytokines levels, kidney and liver functionality, as well as lung histopathological status in rats submitted to chronic CsA treatment without undergoing any transplantation. Male Wistar rats were divided into three groups. The control group received vehicle (corn oil), and treated groups received CsA 5 or 15 mg/kg, by daily gastric gavage during 8 weeks. The results demonstrated that CsA treatment decreases blood levels of interleukins 1α (IL-1α), 1ß (IL-1ß) and interleukin 2 (IL-2), but does not alter interleukin 6 (IL-6) and IFN-γ levels. Serum biochemical markers of renal (creatinine) and hepatic (SGPT and SGOT) injury/dysfunction did not vary with CsA treatment, despite the presence of small histological alterations, suggesting that the function of these metabolic organs were preserved. Pulmonary histopathological lesions were observed in the CsA groups, and they were attributed to the activation of the local immunoresponse mechanisms by the normal microbiota in immunosuppressive CsA cases. These results suggest that the CsA concentrations administered in our experimental protocol were able to induce immunosuppression in rats without causing nephro and hepatotoxicity.

Documentação fotográfica intra-operatória / Intraoperative photographic documentation

O prontuário médico é um dever ético, sendo a documentação fotográfica uma extensão deste, e muitas vezes de grande importância, principalmente na Cirurgia Plástica. A documentação fotográfica pré-operatória e pós-operatória em Cirurgia Plástica é rotineiramente utilizada, enquanto a fotografia intra-operatória ainda não, apesar de facilitar e auxiliaras divulgações científicas e ser uma forte aliada nos registros médicos. O presente artigo apresenta uma forma simples, de baixo custo, capaz de permitir a documentação fotográfica intra-operatória com ótima qualidade, preservando-se o ângulo de visão do próprio cirurgião, com menores riscos de contaminação, demandando menor tempo de interrupção e independente de auxiliares externos. Tal ferramenta consiste na associação de uma capa à prova de água à câmera fotográfica digital.

The medical records are an ethic obligation and the photographic documentation is part of it; many times with huge importance, mainly in Plastic Surgery. The preoperative and postoperative photographic documentation are commonly used in Plastic Surgery, while the intraoperative photograph is not yet, although it helps and facilitates scientific reports and it is a strong ally in medical records. The present article shows a simple way of taking intraoperative photography documentation, with high quality, low cost, preserving the surgeons angle of vision, with low risk of contamination, demanding less time of interruption and independent of external. This tool consists simply the association of a waterproof case with the digital camera.

Efeitos cardiovascular e metabólico da reposição volêmica com solução salina hipertônica 7,5 por cento associada ou não ao hidroxietilamido 130/0,4 em gatas com hipovolemia induzida / Cardiovascular and metabolic effects of volemic expansion with 7.5 percent hypertonic saline alone or in combination with hydroxyethylstarch 130/0.4 in cats with induced hypovolemia

A hipovolemia é caracterizada por uma perda de fluido corpóreo, cursando com inadequado fluxo circulatório e consequentemente lesão tecidual. Neste trabalho, objetivou-se comparar a expansão volêmica resultante da administração de solução salina hipertônica (NaCl 7,5 por cento), isolada ou em associação com hidroxietilamido 130/0,4 (HES 130/0,4), em gatas com hipovolemia induzida, sob anestesia geral inalatória com isofluorano. Foram utilizadas 12 gatas, sem raça definida, adultas, com massa corporal média de 3,07±0,56kg. Os animais foram anestesiados com isofluorano e, após a preparação cirúrgica, foram mantidos em 1CAM sob ventilação controlada. Após a estabilização do plano anestésico, foram avaliados os parâmetros basais. Em ato contínuo, iniciou-se a fase de hipovolemia, por meio da retirada de 30ml kg-1 de sangue da artéria femoral. Após 60 minutos da estabilização do quadro de hipovolemia, foi realizada nova mensuração dos dados, alocando-se os animais aleatoriamente em dois grupos: GSH (grupo solução hipertônica, n=6), que receberam, na fase de expansão volêmica, NaCl 7,5 por cento isolada, na dose de 4ml kg-1, e GSHC (grupo salina hipertônica associado ao coloide, n=6), que receberam NaCl 7,5 por cento, na mesma dose citada, em associação com HES 130/0,4, na dose de 30ml kg-1. Após realização do tratamento, foram avaliados novamente os efeitos cardiovasculares e hemogasométricos por 120 minutos. As pressões arteriais média (PAM), sistólica (PAS) e diastólica (PAD) foram maiores logo após a expansão volêmica (T0) para o GSH. De T45 até T120, as PAM, PAS e PAD foram maiores para o GSHC, em comparação com o GSH. A pressão venosa central foi maior no GSHC até T60. Não foram observadas diferenças entre grupos para frequência cardíaca e respiratória, íons sódio e potássio, déficit de base, bicarbonato, saturação de oxigênio na hemoglobina, glicose, PaCO2, PaO2 e pH. Conclui-se que a administração de NaCl 7,5 por cento isoladamente aumenta rapidamente a PAM, PAS e PAD em gatos com hipovolemia induzida, mantendo esse efeito por apenas 30 minutos, enquanto que a administração de hidroxietilamido 130/0,4 associado à NaCl 7,5 por cento promove reestabelecimento mais tardio (após 30 minutos), porém mais duradouro (até 120 minutos) da PAM, PAS e PAD em gatas com hipovolemia induzida. A administração de HES 130/0,4 associada à NaCl 7,5 por cento promove aumento acentuado da PVC por até 60 minutos após a administração.

Hypovolemia is characterized by body fluid loss leading to an inadequate circulatory flow and consequently tissue injury. The aim of this study was to compare the volume expansion using hypertonic saline solution (NaCl 7.5 percent) alone or in combination with hydroxyethylstarch 130/0.4 (HES 130/0.4) in cats under general anesthesia with isoflurane and exposed to experimental hypovolemia. Twelve adult mixed breed cats, average weight of 3.07±0.56kg were used in this study. The queens were anesthetized using isoflurane and after surgical prepare, anesthesia were maintained with 1 MAC of isoflurane and the animals were mechanically ventilated. After waiting for anesthesia stabilization, baseline cardiovascular and hemogasometric parameters were recorded. Hypovolemia was induced by withdrawing 30mL kg-1 of blood from the femoral artery, after an hour, data were reassessed, and then, the animals were allocated into two groups: HSG (hypertonic saline group, n=6), which received 4mL kg-1 of 7.5 percent NaCl, and HSCG (hypertonic saline, associated to HES 130/0,4 group n=6) which received 4mL kg-1 of 7.5 percent NaCl associated to 30ml kg-1 of HES 130/0.4. After the treatments, cardiovascular and hemogasometric parameters were assessed for 120 minutes. Systolic blood pressure (SAP), diastolic blood pressure (DAP), mean arterial pressure (MAP) were significantly higher after volemic expansion on HSG at T0. From T45 to T120, SAP, DAP and MAP were significantly higher in HSCG when compared to HSG. Central venous pressure was higher in HSCG up to T60. There were no significant differences between groups on heart rate, respiratory rate, Na+, K+, base excess, bicarbonate, hemoglobin saturation, glucose, PaCO2, PaO2 and pH. The administration of 7.5 percent NaCl, promoted a faster increase in MAP, SAP and DAP in cats with induced hypovolemia and these effects were maintained for 30 minutes while the administration of HES 130/0.4 in combination with 7.5 percent NaCl promoted a delayed reestablishment (30 minutes after treatment) of MAP, SAP and DAP in cats with induced hypovolemia, but lasting up to T120. The combination of HES 130/0.4 and NaCl 7.5 percent promoted a significant increase on CVP for up to 60 minutes after the treatment.

Chronic treatment with cyclosporine affects systemic purinergic parameters, homocysteine levels and vascular disturbances in rats.

Vascular disease is a major cause of morbidity and mortality among transplanted recipients and cyclosporine (CsA) treatment has been consistently implicated in this event. In this study we assessed total blood homocysteine levels (tHcy), ecto-nucleotidase activities and adenine nucleotide/nucleoside levels searching for parameters related to the mechanisms of vascular damage induced by chronic CsA treatment in non-transplanted rats. Thirty male Wistar rats were divided in three groups: control group treated with corn oil, CsA 5mg/kg and CsA 15 mg/kg, administered by daily gastric gavage during 8 weeks. CsA 15 mg/kg treatment increased blood levels of tHcy. Both CsA treatments (5mg/kg and 15 mg/kg) decreased adenine nucleotides hydrolysis by ecto-nucleotidases in serum, which negatively correlated with tHcy levels (r: -0.74, r: -0.63 and r: -0.63, p<0.004, for ATP, ADP and AMP, respectively). CsA 15mg/kg induced a statistically significant increase in ADP and decrease in adenosine (ADO) plasma levels compared to control group. THcy levels were positively correlated with plasma ADP levels and negatively correlated with ADO levels (r: 0.84, p<0.0001 and r: -0.68, p<0.0001, respectively). Rats under CsA 15 mg/kg treatment presented cell injury and inflammatory responses in the endothelium and intima layer of the aorta artery. In conclusion, blood ecto-nucleotidases activity, tHcy, and ADP and ADO levels may be implicated in vascular injury induced by CsA treatment.

Associação de cetamina S(+) e midazolam pelo método convencional de cálculo e pela extrapolação alométrica em bugios-ruivo (Alouatta guariba clamitans): resposta clínica e cardiorrespiratória / S(+) ketamine and midazolam association by the conventional method of calculation and allometric extrapolation in red howler monkeys (Alouatta guariba clamitans): clinical and cardiopulmonary response

O objetivo deste estudo foi avaliar o protocolo de contenção química com cetamina S(+) e midazolam em bugios-ruivos, comparando o cálculo de doses pelo método convencional e o método de extrapolação alométrica. Foram utilizados 12 macacos bugios (Alouatta guariba clamitans) hígidos, com peso médio de 4,84±0,97kg, de ambos os sexos. Após jejum alimentar de 12 horas e hídrico de seis horas, realizou-se contenção física manual e aferiram-se os seguintes parâmetros: frequência cardíaca (FC), frequência respiratória (f), tempo de preenchimento capilar (TPC), temperatura retal (TR), pressão arterial sistólica não invasiva (PANI) e valores de hemogasometria arterial. Posteriormente, os animais foram alocados em dois grupos: GC (Grupo Convencional, n=06), os quais receberam cetamina S(+) (5mg kg-1) e midazolam (0,5mg kg-1), pela via intramuscular, com doses calculadas pelo método convencional; e GA (Grupo Alometria, n=06), os quais receberam o mesmo protocolo, pela mesma via, utilizando-se as doses calculadas pelo método de extrapolação alométrica. Os parâmetros descritos foram mensurados novamente nos seguintes momentos: M5, M10, M20 e M30 (cinco, 10, 20 e 30 minutos após a administração dos fármacos, respectivamente). Também foram avaliados: qualidade de miorrelaxamento, reflexo podal e caudal, pinçamento interdigital, tempo para indução de decúbito, tempo hábil de sedação, qualidade de sedação, e tempo e qualidade de recuperação. O GA apresentou menor tempo para indução ao decúbito, maior grau e tempo de sedação, bem como redução significativa da FC e PANI de M5 até M30, quando comparado ao GC. Conclui-se que o grupo no qual o cálculo de dose foi realizado por meio da alometria (GA) apresentou melhor grau de relaxamento muscular e sedação, sem produzir depressão cardiorrespiratória significativa.

The aim of this study was to evaluate a protocol of chemical restraint comparing the conventional method of calculation (weight dose) and allometric extrapolation. Twelve healthy red howler monkeys (Alouatta guariba clamitans), average weight 4.84±0.97kg, male and female, were used for this study. After a 12-hour period of food restriction and 6 hours of water restriction, the animals were physically restraint and the following parameters were measured: heart rate (HR), respiratory rate (RR), capillary refill time (CRT), rectal temperature (RT), non invasive systolic arterial pressure (NISAP) and arterial blood gases analysis. The animals were distributed into two groups: CG (Conventional Group, n=6), in which the animals received S(+) ketamine (5mg kg-1) and midazolam (0.5mg kg-1), by intramuscular (IM) injection; and AG (Allometry Group, n=6), in which the animals also received S(+) ketamine and midazolan IM, but the doses were calculated by allometric extrapolation. Parameters were evaluated at the following moments: M5, M10, M20 and M30 (5, 10, 20 and 30 minutes after IM injection, respectively). Muscle relaxation, pedal and caudal reflexes, interdigital pinch, recumbency time, sedation's quality and duration, and recovery time and its quality were also evaluated. The AG had a faster time for recumbency, higher period and quality of sedation, and a significantly reduction on HR and SAP from M5 to M30 when compared to CG. It was concluded that allometric extrapolation presented a better muscle relaxation and sedation without significant cardiorespiratory depression.