Desenvolvimento e validação de testes de fluxo lateral para a detecção de cultivares geneticamente modificados e de aflatoxinas em produtos agrícolas

The expansion of cultivated areas with genetically modified crops (GM) is a worldwide phenomenon, stimulating regulatory authorities to implement strict procedures to monitor and verify the presence of GM varieties in agricultural crops. With the constant growing of plant cultivating areas all over the world, consumption of aflatoxin-contaminated food also increased. Aflatoxins correspond to a class of highly toxic contaminants found in agricultural products that can have harmful effects on human and animal health. Therefore, the safety and quality evaluation of agricultural products are important issues for consumers. Lateral flow tests (strip tests) is a promising method for the detection both proteins expressed in GM crops and aflatoxins-contaminated food samples. The advantages of this technique include its simplicity, rapidity and cost-effective when compared to the conventional methods. In this study, two novel and sensitive strip tests assay were developed for the identification of: (i) Cry1Ac and Cry8Ka5 proteins expressed in GM cotton crops and; (ii) aflatoxins from agricultural products. The first strip test was developed using a sandwhich format, while the second one was developed using a competitive format. Gold colloidal nanoparticles were used as detector reagent when coated with monoclonal antibodies. An anti-species specific antibody was sprayed at the nitrocellulose membrane to be used as a control line. To validate the first strip test, GM (Bollgard I® e Planta 50- EMBRAPA) and non-GM cotton leaf (Cooker 312) were used. The results showed that the strip containing antibodies for the identification of Cry1Ac and Cry8Ka5 proteins was capable of correctly distinguishing between GM samples (positive result) and non-GM samples (negative result), in a high sensitivity manner. To validate the second strip test, artificially contaminated soybean with Aspergillus flavus (aflatoxin-producing fungus) was employed. Food samples, such as milk and soybean, were also evaluated for the presence of aflatoxins. The strip test was capable to distinguish between samples with and without aflatoxins samples, at a sensitivity concentration of 0,5 μg/Kg. Therefore, these results suggest that the strip tests developed in this study can be a potential tool as a rapid and cost-effective method for detection of insect resistant GM crops expressing Cry1Ac and Cry8Ka5 and aflatoxins from food samples.

Desenvolvimento e validação de testes de fluxo lateral para a detecção de cultivares geneticamente modificados e de aflatoxinas em produtos agrícolas

The expansion of cultivated areas with genetically modified crops (GM) is a worldwide phenomenon, stimulating regulatory authorities to implement strict procedures to monitor and verify the presence of GM varieties in agricultural crops. With the constant growing of plant cultivating areas all over the world, consumption of aflatoxin-contaminated food also increased. Aflatoxins correspond to a class of highly toxic contaminants found in agricultural products that can have harmful effects on human and animal health. Therefore, the safety and quality evaluation of agricultural products are important issues for consumers. Lateral flow tests (strip tests) is a promising method for the detection both proteins expressed in GM crops and aflatoxins-contaminated food samples. The advantages of this technique include its simplicity, rapidity and cost-effective when compared to the conventional methods. In this study, two novel and sensitive strip tests assay were developed for the identification of: (i) Cry1Ac and Cry8Ka5 proteins expressed in GM cotton crops and; (ii) aflatoxins from agricultural products. The first strip test was developed using a sandwhich format, while the second one was developed using a competitive format. Gold colloidal nanoparticles were used as detector reagent when coated with monoclonal antibodies. An anti-species specific antibody was sprayed at the nitrocellulose membrane to be used as a control line. To validate the first strip test, GM (Bollgard I® e Planta 50- EMBRAPA) and non-GM cotton leaf (Cooker 312) were used. The results showed that the strip containing antibodies for the identification of Cry1Ac and Cry8Ka5 proteins was capable of correctly distinguishing between GM samples (positive result) and non-GM samples (negative result), in a high sensitivity manner. To validate the second strip test, artificially contaminated soybean with Aspergillus flavus (aflatoxin-producing fungus) was employed. Food samples, such as milk and soybean, were also evaluated for the presence of aflatoxins. The strip test was capable to distinguish between samples with and without aflatoxins samples, at a sensitivity concentration of 0,5 μg/Kg. Therefore, these results suggest that the strip tests developed in this study can be a potential tool as a rapid and cost-effective method for detection of insect resistant GM crops expressing Cry1Ac and Cry8Ka5 and aflatoxins from food samples.

Preferência e competição alimentar em um grupo de Sapajus flavius em fragmento de Mata Atlântica em Caaporã – Paraíba – Brasil

Feeding is the primary selective pressure in all forms of animals. Nutritional ecological models predict consequences of preferred and non-preferred food consumption on behavioural, physiological and morphological adaptations. At same time, socioecological models infer socio-organizarion patterns based on feeding competition faced by animals. A list of preferred foods, and inferences regarding the intensity of feeding competition and its behavioural consequences are information of much importance for management of populations in fragments. In this work we observed the feeding behavior and spatial positioning of a group of more than 100 blond capuchin monkeys (Sapajus flavius) that inhabit a fragment of Atlantic forest, surrounded by sugarcane plantation. We compared the consumption of different food items with their monthly availability in the area to define the preferred and fallback food items. We recorded the vocalizations of aggression and the inter-individual distance (area of Minimum Convex Polygon/n individuals) to infer the type of food competition experienced by animals. In the year studied the fruit feeding time correlated with top consumed fruit productivity, indicating preference for fruits. Our data indicate that the species Elaeis sp., Cecropia palmata, Inga spp. and Simarouba amara are the preferred food items in the diet. Available all year round and uniformly distributed, sugarcane was a regular item in the diet and its was characterized as a staple fallback food for this group. Although fruits are preferential food items, direct competition rate did not correlate to fruit productivity in the area, maintaining the high rates throughout the year (2.45 events/ hour). The inter-individual distance index positively correlated with rain fall indicating scramble food competition. The number of neighbours of females carrying infants was smaller when fruit productivity is low, indicating that females carrying infants are suffering increased indirect competition. Our data indicates that blond capuchins in this fragment make use of sugar cane as a staple fallback food, which evidence the importance of sugar cane landscape for the survival of this critically endangered capuchin species in fragmented habitats in Northeast Brazil. A preliminary list of preferred and important foods is offered, and can assist in the choice of trees for reforestation, better fragments to be preserved and areas of release and translocation of animals. We did not observe an increase of contest competition while using preferred foods, but when using staple FBF. This may be due the altered environment, which results in high competition food throughout the year. Both the food preference as the social and behavioral consequences of high food competition experienced by animals in this fragment must be accompanied over the years to ensure the survival of this population.

Preferência e competição alimentar em um grupo de Sapajus flavius em fragmento de Mata Atlântica em Caaporã – Paraíba – Brasil

Feeding is the primary selective pressure in all forms of animals. Nutritional ecological models predict consequences of preferred and non-preferred food consumption on behavioural, physiological and morphological adaptations. At same time, socioecological models infer socio-organizarion patterns based on feeding competition faced by animals. A list of preferred foods, and inferences regarding the intensity of feeding competition and its behavioural consequences are information of much importance for management of populations in fragments. In this work we observed the feeding behavior and spatial positioning of a group of more than 100 blond capuchin monkeys (Sapajus flavius) that inhabit a fragment of Atlantic forest, surrounded by sugarcane plantation. We compared the consumption of different food items with their monthly availability in the area to define the preferred and fallback food items. We recorded the vocalizations of aggression and the inter-individual distance (area of Minimum Convex Polygon/n individuals) to infer the type of food competition experienced by animals. In the year studied the fruit feeding time correlated with top consumed fruit productivity, indicating preference for fruits. Our data indicate that the species Elaeis sp., Cecropia palmata, Inga spp. and Simarouba amara are the preferred food items in the diet. Available all year round and uniformly distributed, sugarcane was a regular item in the diet and its was characterized as a staple fallback food for this group. Although fruits are preferential food items, direct competition rate did not correlate to fruit productivity in the area, maintaining the high rates throughout the year (2.45 events/ hour). The inter-individual distance index positively correlated with rain fall indicating scramble food competition. The number of neighbours of females carrying infants was smaller when fruit productivity is low, indicating that females carrying infants are suffering increased indirect competition. Our data indicates that blond capuchins in this fragment make use of sugar cane as a staple fallback food, which evidence the importance of sugar cane landscape for the survival of this critically endangered capuchin species in fragmented habitats in Northeast Brazil. A preliminary list of preferred and important foods is offered, and can assist in the choice of trees for reforestation, better fragments to be preserved and areas of release and translocation of animals. We did not observe an increase of contest competition while using preferred foods, but when using staple FBF. This may be due the altered environment, which results in high competition food throughout the year. Both the food preference as the social and behavioral consequences of high food competition experienced by animals in this fragment must be accompanied over the years to ensure the survival of this population.

Praias urbanas e racionalidade ambiental: avaliação da gestão, das intervenções urbanísticas e dos usos na praia de ponta negra

This dissertation addresses issues considered essential for sustainable development of urban waterfronts and beaches. Many of these spaces, even though they are of public authorities, economic market and general population interests - due to its landscape, its importance for recreation purposes and as a basis of " Sun and beach Tourism " (Turismo de Sol e Praia), among other factors - have shown aesthetic, health and cultural degradation, entailing environmental, economic and social losses and conflicts. Based on this perception, the research aims to understand the main reasons for these negative results for beach spaces. To this end, it was chosen the case study of a typical urban beach, Ponta Negra Beach, located in Natal, RN. Ponta Negra is associated with the "postcard" of the city and it has been deserving of municipal urban planning legislation that legally recognizes the importance of its landscape. Also it has received constant investments by the Government through urban projects, arguing to leave the site attractive to its users. Nevertheless, in the last fifteen years, the beach has lived with the expansion of its problems, such as those related to bathing water, to coastal erosion, and to the mangling of its natural surroundings. Social conflicts have also been frequent in this time frame: conflicts between residents of the waterfront and traders who work on the beach, between the traders themselves, between the managers of space and fishermen, between managers and formal and informal traders. Many of these social and environmental conflicts have taken such grand proportions that became legal matters. Assuming that the problems identified are related to the issue of rationality - understood as a system of values, norms and actions that relate means and ends - and upholding the need for focused research on "environmental rationality" to understand and interpret the dynamics of social and environmental problems encountered on site, the research that guides the study relies on the Mexican economist Enrique Leff's theory on "environmental rationality" which, briefly, can be defined as a system of values, norms, actions and means and ends relations based on the principles of environmental management and sustainable development. Among other aspects, rationality encompasses cross-sectional planning of public administration, the participation of society in the management of environmental resources, interdisciplinary reorganization of knowledge, the clash of opposing interests and the conciliation of common goals of different social actors. The study evaluates the relationship between "environmental rationality", as proposed by Enrique Leff, with the management, urban interventions and uses observed in Ponta Negra Beach. For that, some benchmarks were established and considered in the research as related to sustainable development of the "beachy" atmosphere. Analytical instruments chosen were the urban transformations and the environmental and social problems that have been the target of lawsuits. Also part of the study, the problems that were the subject of civil investigations, which are investigation procedures carried out by the Prosecutor's Office.

Praias urbanas e racionalidade ambiental: avaliação da gestão, das intervenções urbanísticas e dos usos na praia de ponta negra

This dissertation addresses issues considered essential for sustainable development of urban waterfronts and beaches. Many of these spaces, even though they are of public authorities, economic market and general population interests - due to its landscape, its importance for recreation purposes and as a basis of " Sun and beach Tourism " (Turismo de Sol e Praia), among other factors - have shown aesthetic, health and cultural degradation, entailing environmental, economic and social losses and conflicts. Based on this perception, the research aims to understand the main reasons for these negative results for beach spaces. To this end, it was chosen the case study of a typical urban beach, Ponta Negra Beach, located in Natal, RN. Ponta Negra is associated with the "postcard" of the city and it has been deserving of municipal urban planning legislation that legally recognizes the importance of its landscape. Also it has received constant investments by the Government through urban projects, arguing to leave the site attractive to its users. Nevertheless, in the last fifteen years, the beach has lived with the expansion of its problems, such as those related to bathing water, to coastal erosion, and to the mangling of its natural surroundings. Social conflicts have also been frequent in this time frame: conflicts between residents of the waterfront and traders who work on the beach, between the traders themselves, between the managers of space and fishermen, between managers and formal and informal traders. Many of these social and environmental conflicts have taken such grand proportions that became legal matters. Assuming that the problems identified are related to the issue of rationality - understood as a system of values, norms and actions that relate means and ends - and upholding the need for focused research on "environmental rationality" to understand and interpret the dynamics of social and environmental problems encountered on site, the research that guides the study relies on the Mexican economist Enrique Leff's theory on "environmental rationality" which, briefly, can be defined as a system of values, norms, actions and means and ends relations based on the principles of environmental management and sustainable development. Among other aspects, rationality encompasses cross-sectional planning of public administration, the participation of society in the management of environmental resources, interdisciplinary reorganization of knowledge, the clash of opposing interests and the conciliation of common goals of different social actors. The study evaluates the relationship between "environmental rationality", as proposed by Enrique Leff, with the management, urban interventions and uses observed in Ponta Negra Beach. For that, some benchmarks were established and considered in the research as related to sustainable development of the "beachy" atmosphere. Analytical instruments chosen were the urban transformations and the environmental and social problems that have been the target of lawsuits. Also part of the study, the problems that were the subject of civil investigations, which are investigation procedures carried out by the Prosecutor's Office.

Efeitos in vivo e in vitro de agonistas parciais do receptor NOP: implicações para o tratamento da ansiedade, depressão e mania

Introduction: This study aimed to investigate the effects of the two peptide NOP partial agonists (UFP-113 and [F/G]N/OFQ(1-13)NH2) and the non peptide NOP partial agonist (AT-090) in the mouse emotional behavior as well as in the intracellular transduction pathways following the receptor binding. Methods: Male Swiss or CD-1 mice were used in this study together with NOP(+/+) and NOP(-/-) mice. The elevated plus maze (EPM) was used to evaluate the effects of compounds on anxiety-like behaviors. Diazepam and the NOP agonists, N/OFQ and Ro 65-6570, were used as positive controls in the EPM. NOP(+/+) and NOP(-/-) mice were used to evaluate the selectivity of those compounds that induced anxiolytic-like behaviors. The forced swim test (FST) was used to evaluate the effects of compounds on depressive-like behaviors. Nortriptyline and the NOP antagonists, UFP-101 and SB-612111, were used as positive controls in the FST. The effects of N/OFQ, UFP-101, SB-612111, UFP-113, [F/G]N/OFQ(1-13)NH2, and AT-090 were assessed in the methylphenidate-induced hyperlocomotion (MIH) test; in this assay valproate was used as positive control. The G protein and β-arrestin 2 transduction pathways of NOP receptor agonists (N/OFQ and Ro 65-6570), antagonist (UFP-101), and partial agonists (UFP-113, [F/G]N/OFQ(1-13)NH2, and AT-090) were also evaluated using an innovative assay that measures a bioluminescence resonance energy transfer process. For this, cell lines permanently co-expressing the NOP receptor coupled to luciferase (energy donor), and green fluorescent protein (energy acceptor) coupled to one of the effector proteins (G protein or β-arrestin 2) were used. Results: Diazepam (1 mg/kg), N/OFQ (1 nmol), Ro 65-6570 (0.1 mg/kg), and AT-090 (0.01 mg/kg) induced anxiolytic-like effect in mice in the EPM. The effects of Ro 65-6570 and AT-090 were selective to NOP receptor. UFP-113 (0.01-1 nmol) and [F/G]N/OFQ(1-13)NH2 (0.1-3 nmol) were inactive in the EPM. In the FST, nortriptyline (30 mg/kg), UFP-101 (10 nmol), SB-612111 (10 mg/kg), UFP-113 (0.01 and 0.1 nmol), and [F/G]N/OFQ(1-13)NH2 (0.3 and 1 nmol) induced antidepressant-like effects, while AT-090 (0.001-0.1 mg/kg) was inactive in this assay. The effects of UFP-113 and [F/G]N/OFQ(1-13)NH2 were selective to NOP receptor. Valproate (400 mg/kg) counteracted methylphenidate (MPH, 10 mg/kg)-induced hyperlocomotion in mice in the open field. N/OFQ (1 nmol), UFP-113 (0.01-0.1 nmol), and [F/G]N/OFQ(1-13)NH2 (1 nmol) were also able to reduce the MPH-induced hyperlocomotion, without changing the locomotor activity per se. The effect of UFP-113 was selective to NOP receptor. The UFP-101 (10 nmol), SB-612111 (10 mg/kg), and AT-090 (0.001-0.03 mg/kg) did not change the hyperlocomotor effect of methylphenidate. In vitro, N/OFQ and Ro 65-6570 behaved as NOP full agonists for G-protein and β-arrestin 2 pathways. AT-090 behaved as NOP receptor partial agonist for both transduction pathways, while UFP-113 and [F/G]N/OFQ(1-13)NH2 behaved as partial agonists and antagonists of NOP receptor for NOP/G protein and NOP/β-arrestin 2, respectively. UFP-101 behaved as NOP receptor antagonist for both transduction pathways. Conclusion: NOP ligands producing same effects on NOP/G protein interaction (partial agonism), but with opposite effects on β-arrestin 2 recruitment (partial agonism vs antagonism), can promote different in vivo effects on anxiety and mood as it was observed in the behavioral tests. This work corroborates the potential of NOP receptor as an innovative pharmacological target for the treatment of emotional disorders.

Efeitos in vivo e in vitro de agonistas parciais do receptor NOP: implicações para o tratamento da ansiedade, depressão e mania

Introduction: This study aimed to investigate the effects of the two peptide NOP partial agonists (UFP-113 and [F/G]N/OFQ(1-13)NH2) and the non peptide NOP partial agonist (AT-090) in the mouse emotional behavior as well as in the intracellular transduction pathways following the receptor binding. Methods: Male Swiss or CD-1 mice were used in this study together with NOP(+/+) and NOP(-/-) mice. The elevated plus maze (EPM) was used to evaluate the effects of compounds on anxiety-like behaviors. Diazepam and the NOP agonists, N/OFQ and Ro 65-6570, were used as positive controls in the EPM. NOP(+/+) and NOP(-/-) mice were used to evaluate the selectivity of those compounds that induced anxiolytic-like behaviors. The forced swim test (FST) was used to evaluate the effects of compounds on depressive-like behaviors. Nortriptyline and the NOP antagonists, UFP-101 and SB-612111, were used as positive controls in the FST. The effects of N/OFQ, UFP-101, SB-612111, UFP-113, [F/G]N/OFQ(1-13)NH2, and AT-090 were assessed in the methylphenidate-induced hyperlocomotion (MIH) test; in this assay valproate was used as positive control. The G protein and β-arrestin 2 transduction pathways of NOP receptor agonists (N/OFQ and Ro 65-6570), antagonist (UFP-101), and partial agonists (UFP-113, [F/G]N/OFQ(1-13)NH2, and AT-090) were also evaluated using an innovative assay that measures a bioluminescence resonance energy transfer process. For this, cell lines permanently co-expressing the NOP receptor coupled to luciferase (energy donor), and green fluorescent protein (energy acceptor) coupled to one of the effector proteins (G protein or β-arrestin 2) were used. Results: Diazepam (1 mg/kg), N/OFQ (1 nmol), Ro 65-6570 (0.1 mg/kg), and AT-090 (0.01 mg/kg) induced anxiolytic-like effect in mice in the EPM. The effects of Ro 65-6570 and AT-090 were selective to NOP receptor. UFP-113 (0.01-1 nmol) and [F/G]N/OFQ(1-13)NH2 (0.1-3 nmol) were inactive in the EPM. In the FST, nortriptyline (30 mg/kg), UFP-101 (10 nmol), SB-612111 (10 mg/kg), UFP-113 (0.01 and 0.1 nmol), and [F/G]N/OFQ(1-13)NH2 (0.3 and 1 nmol) induced antidepressant-like effects, while AT-090 (0.001-0.1 mg/kg) was inactive in this assay. The effects of UFP-113 and [F/G]N/OFQ(1-13)NH2 were selective to NOP receptor. Valproate (400 mg/kg) counteracted methylphenidate (MPH, 10 mg/kg)-induced hyperlocomotion in mice in the open field. N/OFQ (1 nmol), UFP-113 (0.01-0.1 nmol), and [F/G]N/OFQ(1-13)NH2 (1 nmol) were also able to reduce the MPH-induced hyperlocomotion, without changing the locomotor activity per se. The effect of UFP-113 was selective to NOP receptor. The UFP-101 (10 nmol), SB-612111 (10 mg/kg), and AT-090 (0.001-0.03 mg/kg) did not change the hyperlocomotor effect of methylphenidate. In vitro, N/OFQ and Ro 65-6570 behaved as NOP full agonists for G-protein and β-arrestin 2 pathways. AT-090 behaved as NOP receptor partial agonist for both transduction pathways, while UFP-113 and [F/G]N/OFQ(1-13)NH2 behaved as partial agonists and antagonists of NOP receptor for NOP/G protein and NOP/β-arrestin 2, respectively. UFP-101 behaved as NOP receptor antagonist for both transduction pathways. Conclusion: NOP ligands producing same effects on NOP/G protein interaction (partial agonism), but with opposite effects on β-arrestin 2 recruitment (partial agonism vs antagonism), can promote different in vivo effects on anxiety and mood as it was observed in the behavioral tests. This work corroborates the potential of NOP receptor as an innovative pharmacological target for the treatment of emotional disorders.

Reserva extrativista Acaú-Goiana: tragédia comum ou decisão coletiva?

In recent years we have observed, in Brazil, the expansion of democratic spaces, providing the use of participatory management strategies in common-pool areas. However, the existence of a co-management model does not guarantee its practice, neither that it will prevent the depletion of common features characterizing the tragedy proposed by Hardin. In this work we analysed the management of Acaú-Goiana RESEX which is located in the cities of Acaú (PB) and Goiana (PE), eight years after its legal creation. We evaluated how much the users of RESEX possess information (even if non formalized) regarding the common use of area and whether they exhibit collectivist values. We used as an interpretative tool the facilitators principles of common resource management (PGRC) identified by Elinor Ostrom and the questionnaire of human values proposed by Schwartz. The human values questionnaire was applied to 240 individuals, half beneficiary and the other half non-beneficiary, only the beneficiaries were submitted to the questionnaire of PGRC. Interviews were conducted in the period of May to September 2014. We tested three hypotheses: 1) The degree of dependence from RESEX resources will play an important role on indirect knowledge of the PGRC; 2) the inclusion of individuals in a communal area (RESEX) increases collectivists values; 3) higher collectivist value rates increases knowledge of PGRC. To this end, we defined levels of socio-economic dependence of RESEX resources, knowledge of PGRC and individual values. GLM statistic analysis of mean comparison and correlation were employed. Our results showed that the knowledge on six of the seven items analyzed in the basic principles (PGRC) is still low. The extended land area and the high number of users may be exerting a detrimental effect on the development of a co-management. Contrary to expected, it was not the dependency who influenced knowledge on the PGRC, but the time spent in contact with the RESEX. This indicates that direct contact with the environment, not the dependence of it, that raises awareness about PGRC. According to our hypothesis, individuals with collectivist values showed greater knowledge of PGRC and reduced hours of work within the RESEX, indicating a greater tendency of individuals refrain their use of the common resource. Individualistic values correlated with less knowledge of a PGRC. Among the beneficiaries, individuals with higher individualistic values had higher monthly income, while among non-beneficiaries there was no such correlation, demonstrating the economic advantages of individualism (free-riding) in situations of non-private use of resources. Our data emphasize the importance of guiding the main actors in the development of a co-participatory management in the direction of the basic principles, and to develop collectivist values among users of a common good can raise awareness of these principles.

Um dermatam sulfato antitrombótico do camarão Litopenaeus vanammei inibe a inflamação e angiogênese

Inflammation is combined of a vascular and a cellular reaction, resulting in different cells and tissue responses, both the intravascular and extravascular environment. As the inflammatory process occurs, coagulation proteases, in particular thrombin (FIIa), are able to initiate various cellular responses in vascular biology and therefore is often observed activation of other biological systems, leading to complications during an event inflammatory, such as thrombosis and angiogenesis. Thus, antagonists molecules of these events are interesting models for the development of novel anti-inflammatory drugs. Thereby, it is worth stressing the glycosaminoglycans (GAGs), which are able to interact with several proteins involved in important biological processes, including inflammation and coagulation. Therefore, this study aimed to evaluate the anti-inflammatory, antithrombotic and anti-angiogenic potentials, as well anticoagulant of a dermatan sulfate-like GAG (DS) extracted from the Litopenaeus vannamei cephalotorax. The compound was obtained after proteolysis and purification by ion-exchange chromatography. After total digestion by DS-like compounds digesting lyases (chondroitinase ABC), the DS-like nature was revealed, and then called DSL. The shrimp compound showed reduced anticoagulant effect by the aPTT assay, but high anti-IIa activity, directly and through heparin cofactor II. On inflammation, the compound had a significant inhibitory effect with the reduction of proinflammatory cytokines. Potential Inhibitory were reported in the antithrombotic and anti-angiogenic assay, the latter being dose dependent. As for anti-hemostatic activity, the polysaccharides did not induced significant bleeding effect. Thus, the results shown by the shrimp DS-like compound indicate this glycosaminoglycan as a biotechnology target with prospects for the development of new multipotent drugs.

Estratégias tróficas do peixe traíra, Hoplias malabaricus e do peixe cascudo, Hypostomus pusarum do açude Marechal Dutra, Rio Grande do Norte, Brasil

The fish, Hoplias malabaricus (Osteichthyes: Erythrinidae) and armored catfish, Hypostomus pusarum (Loricariidae) are of freshwater origin from the neotropical region, and are considered ecologically and economically of important. This work investigated the trophic strategies, the morphology and histology of the digestive tract of these fish captured from the Marechal Dutra reservoir, Acari, the semiarid region of Rio Grande do Norte, Brazil. A total of 133 individuals of H. malabaricus and 118 specimens of H. pusarum were analyzed. The two study species occupy different levels in the food chain and spatial distribution in the water column of the reservoir. The results of this study are presented in the form of four scientific papers. The first article describes the morphology and histology of the digestive tract and the feeding habits of H. malabaricus (Bloch, 1794). This fish has a short intestine, with an intestinal coefficient of 0.72 ± 0.09. The dietary importance index indicates that H. malabaricus feeds preferentially on animal matter, especially on fish (72.8%) and prawns (27.2%). The histology of its digestive tract confirms the carnivorous feeding habit. The second article discusses about the pioneering work on the feeding strategy and the characterization of anatomy and histology of the digestive tract of H. pusarum. The intestine of this fish is long, with an intestinal coefficient of 10.8±0.7. The dietary importance index indicates that H. pusarum feeds preferentially on organic matter in decomposition (88.7%) and on filamentous microalgae and diatoms (11.3%). The third article compares the morphological aspects of the digestive tract of H. malabaricus and H. pusarum, in relation to their food habits. The arrangement of the digestive organs in both species is directly related to the shape of the peritoneal cavity and the form of the body. The short intestine of H. malabaricus and the long intestine of H. pusarum are associated with their feeding habits. The morphology of the digestive tracts of H. malabaricus and H. pusarum confirm their food habits, carnivorous and detritivorous / herbivorous, respectively. The fourth article discusses the food and reproductive aspects (length and weight, length-weight relationship, type of growth and sex ratio) of H. pusarum. This species has a negatively allometric growth, with the predominance of females in the sampled population. H. malabaricus inhabits the pelagic environment and is a carnivore, while H. pusarum, lives in the benthic environment and is characterized as a detritivore/herbivore. Each species studied shows a very different diet, without trophic competition between them. The morphological and anatomical structures of the digestive tract reflect their feeding strategy.

Infecções por enterobacteriaceae resistente aos carbapenêmicos em hospital de ensino: epidemiologia e caracterização molecular

Introduction: The production of KPC (Klebsiella pneumoniae carbapenemase) has become an important mechanism of carbapenem-resistance among Enterobacteriaceae strains. In Brazil, KPC is already widespread and its incidence has increased significantly, reducing treatment options. The “perfect storm” combination of the absence of new drug developmentand the emergence of multidrug-resistant strains resulted in the need for the use of older drugs, with greater toxicity, such as polymyxins. Aims: To determine the occurrence of carbapenemase-producing strains in carbapenem-resistant Enterobacteriaceae isolated from patients with nosocomial infection/colonization during September/2014 to August/2015, to determine the risk factors associated with 30-day- mortality and the impact of inappropriate therapy. Materials and Methods: We performed a case control study to assess the risk factors (comorbidities, invasive procedures and inappropriate antimicrobial therapy) associated with 30-day-mortality, considering the first episode of infection in 111 patients. The resistance genes blaKPC, blaIMP, blaVIM and blaNDM-1 were detected by polymerase chain reaction technique. Molecular typing of the strains involved in the outbreak was performed by pulsed field gel electrophoresis technique. The polymyxin resistance was confirmed by the microdilution broth method. Results: 188 episodes of carbapenem-resistant Enterobacteriaceae infections/colonizations were detected; of these, 122 strains were recovered from the hospital laboratory. The presence of blaKPC gene were confirmed in the majority (74.59%) of these isolates. It was not found the presence of blaIMP , blaVIM and blaNDM-1 genes. K. pneumoniae was the most frequent microorganism (77,13%), primarily responsible for urinary tract infections (21,38%) and infections from patients of the Intensive Care Unit (ICU) (61,38%). Multivariate statistical analysis showed as predictors independently associated with mortality: dialysis and bloodstream infection. The Kaplan-Meier curve showed a lower probability of survival in the group of patients receiving antibiotic therapy inappropriately. Antimicrobial use in adult ICU varied during the study period, but positive correlation between increased incidence of strains and the consumption was not observed. In May and July 2015, the occurrence rates of carbapenem-resistant Enterobacteriaceae KPC-producing per 1000 patient-days were higher than the control limit established, confirming two outbreaks, the first caused by colistin-susceptible KPC-producing K. pneumoniae isolates, with a polyclonal profile and the second by a dominant clone of colistin-resistant (≥ 32 μg/mL) KPC-producing K. pneumoniae. The cross transmission between patients became clear by the temporal and spatial relationships observed in the second outbreak, since some patients occupied the same bed, showing problems in hand hygiene adherence among healthcare workers and inadequate terminal disinfection of environment. The outbreak was contained when the ICU was closed to new admissions. Conclusions: The study showed an endemicity of K. pneumoniae KPC-producing in adult ICU, progressing to an epidemic monoclonal expansion, resulted by a very high antibiotic consumption of carbapenems and polymyxins and facilitated by failures in control measures the unit.

Análise do perfil imunológico e citohistológico durante a transmissão vertical de Trypanosoma cruzi em diferentes estágios da infecção experimental

Trypanosma cruzi is the causative agent of Chagas disease. This trypanosomiasis has become a global public health problem due to migration of Latin Americans to non-endemic countries. In Latin America with the succesful implementation of control domiciliated vector infestation and blood transfusion, the importance of congenital transmission has recently increased. Considering the tight regulation of immune system during gestation, we aimed to investigate the changes in the immune system caused by T.cruzi infection in the gestation outcome. T cruzi G and Y strain were used to infect female BALB/c mice before or after mating with non-infected male mice. The presence of vaginal plug was used as indicative of mating. Females were euthanized 8 days after confirmation of vaginal plug. We used three female control groups, only infected, only infected and non-infected and non-pregnant females. Two groups were infected before mating and other two were infected 4 days after confirmation of vaginal plug. The uterus and spleen were collected to immunochemistry, qPCR, immunofluorescence and cytokine analysis. Our results showed that despite the MMP’s identification being similarly among groups, T.cruzi higher virulent strain can impaire gestation outcome prior mating; the infection also increased cytokines like IFN-γ, IL-1β and IL-4; and leucocytes in uterine environment was altered, responding locally to systemic changes caused by T.cruzi infection. In conclusion this work suggests that T.cruzi infection can impaire gestation outcome and local response to sistemic infection was able to control the infection allowing pregnancy development in some conditions.

Respostas ecofisiológicas de Cybistax antisyphilitica Mart. (Ipê verde) em função das alterações na intensidade de luz

Light varies widely in both time and space in forest formation of “Bioma Cerrado”. Cybistax antisyphilitica occurs in areas typical of this biome, such as cerrado sensu stricto, “cerradões”, and altered areas. The aim of this study was to understand the morphological and physiological responses of C. antisyphilitica to alterations in light intensity. Juvenile plants (5 month of age) were taken to a fragment of semideciduous forest in Uberlândia-MG, and were divided into three treatments: 50 were maintained under the canopy (UC) 20 were kept in small gap (SG) and 20 were maintained under in full sun (FS). The daily courses of chlorophyll a fluorescence were made at the beginning, middle and end of dry season in 2015. At the end of the experiment measurements of chlorophyll content, gas exchange and growth were made. The plants showed dynamic photoinhibition as exhibited by reductions on Fv/Fm close to midday at the end of the dry season. Regarding the effective quantum yield (ΔF/Fm'), plants under FS showed reduced values that coincided with the higher values of electron transport rates (ETR). Plants under FS showed higher values of net CO2 assimilation rates, stomatal conductance, transpiration rates, water use efficiency and chlorophyll content compared to plants under UC. The stem diameter, dry mass of leaves and stem, total dry mass and relative growth rate were higher in plants under FS than plants under UC. On the other hand, plants under UC showed superior values of height, specific leaf area and leaf area ratio. Our results indicate that C. antisyphilitica has plasticity to survive in the contrasting light environments of the semideciduous forests, but this species was able to growth better under full sun conditions.

Manipulação gênica do protozoário Neospora caninum como ferramenta para o estudo das interações parasito-hospedeiro

Neospora caninum is an obligate intracellular parasite classified in the phylum Apicomplexa, characterized by the presence of the apical complex composed by micronemes proteins, rhoptries and dense granules, used by parasite during the adhesion and invasion process of the host cell. This is the mean event in infection pathogenesis generated by N. caninum and other parasites from the phylum Apicomplexa, promoting influence in the parasite biology and the interface between the parasite and its host. Therefore, molecular tools have been developed in order to identify and characterize these possible virulence factors. Thus, the present study sought to establish a specific system of genetic manipulation of N. caninum, searching for the improvement of the genetics manipulation of this parasite. So, we developed genetically depleted N. caninum to Rop9 rhoptry using the pU6-Universal CRISPR-Cas9 plasmid of T. gondii modified by the insertion of Ku80. The Rop9 depleted parasite showed important during initial phase of invasion and replication of the parasite, however it was not characterized as a potential virulence fator for N. caninum. Furthermore, T. gondii proteins were expressed in N. caninum by the use of specific vectors for this parasite, showing an heterologous system for the study of Toxoplasma proteins, due to the fact that Gra15 or Gra24 of type II T. gondii and Rop16 of type I T. gondii were expressed in N. caninum tachyzoites in a stable way and keept its biological phenotype, as already presented the former parasite, that naturaly expresses these proteins. In addition, it was observed that N. caninum induced an inflammasome activation through NLRP3, ASC and Caspase-1. IL-1R/MyD88 demonstrated an indirect pathway in the control of parasite replication. Furthermore, it was observed that this activation is dependent of the potassium efflux and that different strains of N. caninum keep this activation profile. However, T. gondii strains block this activation, making necessary a prior signal in order to active the inflamosome pathway. Type I T. gondii Rop16 was identified as responsible for blocking this activation, in a dependent way to the STAT3 activation. Therefore, the development of molecular tools and their application in N. caninum may prove to be useful to identify and characterize virulent factors involved in the pathogenesis by these two protozoans.