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 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.

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.

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.

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.

Brain lesions in pigs affected with postweaning multisystemic wasting syndrome.

Anti-porcine circovirus type 2 (anti-PCV2) immunostaining was associated with cerebellar lymphohistiocytic vasculitis combined with hemorrhages (50 pigs) or with lymphohistiocytic meningitis (23 pigs) in pigs naturally affected with postweaning multisystemic wasting syndrome (PMWS). The animals originated from 12 farms in Rio Grande do Sul, Brazil. In total, 456 unthrifty 3- to 5-month-old postweaning pigs confirmed as PMWS cases were necropsied. Although most findings mimicked those extensively reported in PMWS-affected pigs, there were distinctive brain lesions that included multiple hemorrhages in the cerebellar leptomeninges associated with lymphohistiocytic vasculitis and fibrinoid degeneration in vessels of the cerebellum and periventricular areas (69 pigs). These vascular lesions were also seen in conjunction with lymphohistiocytic meningitis (38 additional pigs). PCV2 antigen was immunohistochemically demonstrated in the cytoplasm and nuclei from intralesional perivascular macrophages and endothelial-like cells in brain tissues. Together these findings suggest that these lesions were caused by PCV2.

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.

Herd-level risk factors for Neospora caninum seroprevalence in dairy farms in southern Brazil.

A cross-sectional study was used to test the relationship between herd seroprevalence to Neospora caninum and various potential herd-level risk factors in 60 dairy farms located in two distinct regions in southern Brazil. Thirty farms were randomly selected from within each region. A questionnaire was designed to summarize each farm's production system as it might relate to N. caninum transmission. The questionnaire contained 105 closed questions relating to general characteristics of the farms, farm facilities, management, source of food and water, herd health, environment and biosecurity, which included questions relevant to N. caninum transmission, including presence and number of dogs and other animals, purchase of animals and contact with man. Serum samples were collected from 40% of animals in each farm and N. caninum antibodies were detected by immunofluorescent antibody test (IFAT). The association between potential risk factors and the probability of an animal being seropositive was modeled using a generalized estimation equations (GEE) logistic regression model. The model accounted for multilevel correlation of data from multiple animals within herds. The mean (+/-S.D.) number of animals in the 60 herds was 64.5 (+/-45.6), ranging from 20 to 280 females. Blood samples were collected from 1549 animals. The size of the farms varied from 4 to 100 ha (mean 30.1+/-25.9 ha). At least one dog was found in 57 of the 60 dairy farms (95%). The mean number of dogs was 3.1 (+/-1.9), ranging from 0 to 10. All females were raised on pasture. For all cattle sampled, N. caninum seroprevalence was 17.8%. Overall, 93.3% of herds (56/60) had at least one seropositive animal identified. Four variables were significantly associated with N. caninum sero-response in the 57 dairy farms, which were included in the final multivariable model: the number of dogs on the farm, farm area (hectares), feeding pooled sources of colostrum and region. The odds of a cow being seropositive increased 1.13 times for each additional dog present on the farm (P=0.021). Cattle from farms that fed calves colostrum pooled from multiple cows had 1.79 times greater odds for being seropositive for N. caninum (P<0.003). The probability of being seropositive was inverse to the area of the farms, such that cattle had 0.92 times the odds to be seropositive (P=0.014) for each additional 10 ha of farmland. Finally, cattle from farms in region one had 0.71 times the odds to be seropositive than cattle from region two (P=0.035). Results of this study suggest that several risk factors may explain why dairy cattle in Brazil may become exposed to N. caninum. However, further investigation of these factors is necessary because the purpose of this study was to refine and generate hypotheses on N. caninum transmission.

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.

Neurological disorder in dairy cattle associated with consumption of beer residues contaminated with Aspergillus clavatus.

A neurological syndrome in dairy cattle associated with consumption of moldy beer residues is described. The disease occurred on 1 farm in late June 2001, during winter. Six heifers and 1 cow out of 45 cattle were affected during a 3-week period. The affected animals died spontaneously or were euthanized approximately 2-14 days after the onset of clinical signs. The clinical signs were characterized by flaccid paralysis and gait abnormalities. Clinical signs were more pronounced after exercise and included stiff and unsteady gait, knuckling at the fetlocks of the hind limbs, frequent falling, inability to rise, muscular tremors, especially of the head and the hindquarters, and drooling. Main necropsy findings included degenerative and necrotic changes of the larger medial muscle groups of the hindquarters, i.e., adductor, pectineus, quadriceps femoris, rectus femuris, sartorius, semimembranosus, semitendinosus, and vastus medialis, and of the forequarters, including pectoralis descendens, pectoralis ascendens, and transversus pectoralis. The main histologic findings consisted of degenerative and necrotic neuronal changes (chromatolysis) of varying severity and extent affecting selected nuclei of the brainstem and neurons of the ventral horns of the spinal cord. Similar microscopic lesions were observed in the neurons of the spinal cord of 1 experimental sheep force-fed for 35 days with 1 kg/day of the same batch of foodstuff that was originally fed to the cattle. Coarse white or gray lumps, interpreted as mycelia, were observed in the beer by-product. Aspergillus clavatus was the dominant fungus isolated. Deaths ceased after the consumption of beer residue was discontinued. Recovery from illness was observed in 1 animal. The diagnosis was based on epidemiological data, clinical signs, necropsy findings, histological lesions, dosing trial, and mycology. A similar condition caused by consumption of barley by-products, sprouted wheat, corn sprouts, and beetroot screenings contaminated with A. clavatus has been reported in cattle and sheep worldwide.

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.

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.

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.

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.

Porcine circovirus 2 (PCV2) induces a procoagulant state in naturally infected swine and in cultured endothelial cells.

Porcine circovirus 2 (PCV2) is the primary causative agent of porcine circovirus disease (PCVD). PCVD is an emerging disease that has been reported worldwide, associated with wasting, lymphoid depletion, enteritis, pneumonia, vasculitis, ischemic lesions, and necrotizing dermatitis. Although PCVD causes considerable economic losses, the pathogenesis of PCV2 has not been fully understood. The aim of the present work was to study the participation of hemostatic system and of vascular endothelium in PCV2 infection, as well as their possible role in PCVD pathogenesis. Our results showed that naturally PCV2-infected swine displayed a prothrombotic state in vivo, since a diminished coagulation time (recalcification time, activated partial thromboplastin time and prothrombin time), a higher platelet aggregation ability (despite a diminished platelet blood count), and an increased thrombin plasma activity (associated with a reduced fibrinogen level) were observed. The PCV2-infected animals showed vasculitis and positive staining for PCV2 antigen in capillary vessels. Furthermore, PCV2-infected endothelial cells displayed an activated phenotype, characterized by an increase in cell surface procoagulant activity. Moreover, the PCV2-infected endothelial cells pre-treated with exogenous thrombin displayed an increased viral load. This work reports, for the first time, the role of the hemostatic system and of endothelium in the pathogenesis and infectivity of PCV2. The study reinforces the importance of the phenomena which occur during PCV2 infection, and affords a better knowledge of the mechanisms behind the pathophysiology of PCVD.

Toxicity and genotoxicity evaluation of Passiflora alata Curtis (Passifloraceae).

UNLABELLED: Passiflora alata is an official species of Brazilian Pharmacopoeia and its aerial parts are used as medicinal plant by local population as well as constitutes many phytomedicines commercialized in Brazil as sedative. AIMS OF STUDY: To evaluate the acute and sub-acute toxicity and genotoxicity of an aqueous spray-dried extract (PA) of Passiflora alata (2.6% flavonoids). MATERIALS AND METHODS: The acute and the sub-acute toxicity was evaluated in mice and rats, respectively. Behavioural, biochemical, hematological, histological and urine parameters were considered. Genotoxicity was assessed by using micronucleus test performed in peripheral blood and bone marrow cells and comet assay in peripheral blood leukocytes. RESULTS: Mice deaths were not observed up to 4800 mg/kg, p.o., single dose. Rats treated with aqueous extract at dose of 300 mg/kg, p.o., for 14 days did not present biochemical, hematological or histopathological significant alterations when compared to control group. However, these rats showed signs of irritability and did not show weight gain. In addition, mice acutely treated with extract 150, 300 and 600 mg/kg, p.o., presented DNA damage determined by comet assay in peripheral blood cells 3h after treatment. The effect of lower doses (12.5, 25 and 50mg/kg, p.o.) was evaluated at 3, 6 and 24h after treating. Only PA 50mg/kg (p.o.) induced significant damage at 3 and 6h. The maximum damage induction was observed at 6h. When the animals received PA 12.5, 25 or 50mg/kg/day during 3 days (i.e., 72h treatment) DNA damage (comet and micronucleus tests) increased significantly in a dose-dependent manner. CONCLUSION: In conclusion Passiflora alata presented genotoxic effect and deserves further toxicity evaluation in order to guarantee its safety for human use.

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.

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.