Identificação de genes em Chromobacterium violaceum relacionados à resposta ao estresse

The sequencing of the genome of Chromobacterium violaceum identified one single circular chromosome of 4.8 Mb, in which approximately 40% of the founded ORFs are classified as hypothetical conserved or hypothetical. Some genic regions of biotechnological and biological interest had been characterized, e. g., environmental detoxification and DNA repair genes, respectively. Given this fact, the aim of this work was to identify genes of C. violaceum related to stress response, as the ones involved with mechanisms of DNA repair and/or genomic integrity maintenance. For this, a genomic library of C. violaceum was built in Escherichia coli strain DH10B (RecA-), in which clones were tested to UVC resistance, resulting in five candidates clones. In the PLH6A clone were identified four ORFs (CV_3721 to 3724). Two ORFs, CV_3722 and CV_3724, were subcloned and a synergic complementation activity was observed. The occurrence of an operon was confirmed using cDNA from C. violaceum in a RT-PCR assay. Further, it was observed the induction of the operon after the treatment with UVC. Thus, this operon was related to the stress response in C. violaceum. The mutagenesis assay with rifampicin after the treatment with UVC light showed high frequency of mutagenicity for the ORF CV_3722 (Pol III δ subunit). In this way, we propose that the C. violaceum δ subunit can act in DH10B in the translesion synthesis using Pol IV in a RecA independent-manner pathway. In growth curve assays other four clones (PLE1G, PLE7B, PLE10B and PLE12H) were able to complement the function at the dose 5 J/m2 and in mutagenicity assays PLE7B, PLE10B and PLE12H showed frequencies of mutation with significant differences upon the control (DH10B), demonstrating that in some way they are involved with the stress response in C. violaceum. These clones appear to be interrelated, probably regulated by a messenger molecule (eg., nucleotide c-di-GMP) and/or global regulatory molecule (eg., σS subunit of RNA polymerase).The results obtained contribute for a better genetic knowledge of this specie and its response mechanisms to environmental stress.

Identificação de genes em Chromobacterium violaceum relacionados à resposta ao estresse

The sequencing of the genome of Chromobacterium violaceum identified one single circular chromosome of 4.8 Mb, in which approximately 40% of the founded ORFs are classified as hypothetical conserved or hypothetical. Some genic regions of biotechnological and biological interest had been characterized, e. g., environmental detoxification and DNA repair genes, respectively. Given this fact, the aim of this work was to identify genes of C. violaceum related to stress response, as the ones involved with mechanisms of DNA repair and/or genomic integrity maintenance. For this, a genomic library of C. violaceum was built in Escherichia coli strain DH10B (RecA-), in which clones were tested to UVC resistance, resulting in five candidates clones. In the PLH6A clone were identified four ORFs (CV_3721 to 3724). Two ORFs, CV_3722 and CV_3724, were subcloned and a synergic complementation activity was observed. The occurrence of an operon was confirmed using cDNA from C. violaceum in a RT-PCR assay. Further, it was observed the induction of the operon after the treatment with UVC. Thus, this operon was related to the stress response in C. violaceum. The mutagenesis assay with rifampicin after the treatment with UVC light showed high frequency of mutagenicity for the ORF CV_3722 (Pol III δ subunit). In this way, we propose that the C. violaceum δ subunit can act in DH10B in the translesion synthesis using Pol IV in a RecA independent-manner pathway. In growth curve assays other four clones (PLE1G, PLE7B, PLE10B and PLE12H) were able to complement the function at the dose 5 J/m2 and in mutagenicity assays PLE7B, PLE10B and PLE12H showed frequencies of mutation with significant differences upon the control (DH10B), demonstrating that in some way they are involved with the stress response in C. violaceum. These clones appear to be interrelated, probably regulated by a messenger molecule (eg., nucleotide c-di-GMP) and/or global regulatory molecule (eg., σS subunit of RNA polymerase).The results obtained contribute for a better genetic knowledge of this specie and its response mechanisms to environmental stress.

Influência da umidade nas propriedades mecânicas dos compósitos poliméricos híbridos (sisal/ vidro)

This study aims to evaluate the mechanical properties of polymer matrix composites reinforced with sisal fabric bidirectional tissue (Agave sisalana,) and E-glass fibers, containing the following configuration: a polymer matrix hybrid composite (Polyester Resin orthophalic) reinforced with three (3) layers of glass fibers and alternating-2 (two) layers of bidirectional sisal fabric, and finally a composite of polymer matrix reinforced with five (5) layers of glass fiber mat-type E. For this purpose as first step, the preparation of by sisal, since they are not on the market. The composites were made by manual lamination (Hand lay-up) and evaluated for tensile properties and three point bending both in the dry, and wet conditions aswele as immersed in oil. Macroscopic and microscopic characteristics of the materialsweve awalysed, after the completion of the mechanical tests. After the studies, it was proven that the sisal fiber decreases the tensile stiffness of the material above 50% for both situations studied the tensile strength of the material decreases by approximately 40% for the cases mentioned, and when compared to the specific strength stiffness values drop to 14.6% and 29.02% respectively for the dry state only. Constants for bending the values were are to approximately 50% to 25% for strength and stiffness of the material for the cases dry, wet and immersed in oil. Under the influence of tension fluids do not interfere in the stiffness of the material for the bending tests, the same does not occur with the resistance, and these values are modified only in the cases stiffness and flexural strength

Análise de estabilidade de falésias na zona costeira de Baía Formosa - RN

The natural and anthropogenic changes that are occurring at the coastal zone around the world represent the greatest problem to society in this century. This problem becomes more evident due to high density of coastal cities, to growing tourist-estate speculation of those areas and to climate change that tend to trigger and accelerate the erosive processes that operating in the littoral. In this context, it‟s possible perceive ever more a significant increase of problems associated to the coastal erosion and retreat of cliffs in the state of Rio Grande do Norte, where this study area of this search is located. The area is located in the coastal zone of the city of Baía Formosa/RN, in south-eastern coast of Rio Grande do Norte, and has a extension of about 1200 meters along the Porto beach. The main objective of this study is to analyze the stability of these cliffs in this region. Through field investigations, testings and computational analysis using the Finite Element Method and Equilibrium Limit Methods. The area was divided into four sections, and were applied checklists, and also realized characterization tests and direct shear tests with materials obtained along these sections. In this manner, it was found that the segments of cliff in this coastal zone have heights around 4 meters to 14 meters and inclinations of approximately 40° to 90°.However the constituents soils of the cliffs were classified, in general terms in accordance to Unified Soil Classification System (USCS), as clayey sands or silty sands, clays of low plasticity, clayey gravels and poorly graduaded sands. The most variegated and clayey soils belong to base of the cliff. The stability analysis showed that the safety factors ranged in section 01, from 1.92 to 4.93, in the section 02, from 1.00 to 1.43, in the section 03, from 1.36 to 1.75 , and section 04, from 1.00 to 3.64. Thus, the sections 02 and 03 were considered more unstable. However, the section 03 can be considered as the most critical section due to the absence of coastal protection structures and the narrow strip of beach.

Preferência alimentar e identificação das principais fontes de repasto sanguíneo de fêmeas Lutzomyia (Diptera: Psychodidae) em áreas endêmicas para leishmaniose visceral na grande Natal

Leishmaniasis are endemic diseases wild spread in the New and Old World, caused by the flagelated protozoan Leishmania. In the New World, the distribution of different forms of leishmaniasis is mostly in tropical regions. In the State of Rio Grande do Norte, Northeast Brazil, 85% of the captured sand flies fauna is Lutzomyia longipalpis. The distribution of the sand fly vector in the state overlaps with the disease distribution, where the presence of sand flies is associated with presence of animals shelters. The aim of this study was to analyse the blood meal preference of sand flies vector from the genus Lutzomyia spp. in laboratory conditions, to verify the vector life cicle at different temperatures sets and to identify the main blood meal source in endemic areas for visceral leishmaniasis (VL) at peri-urban regions of Natal. Sand flies samples were collected from the municipalities of São Gonçalo do Amarante and Nísia Floresta where female sand flies were grouped for the colony maintenance in the laboratory and for the analysis of the preferred source of sand fly blood meal in natural environment. The prevalence of blood meal preference and oviposition for the females sand flies was 97% for Cavia porcellus with oviposition of 19 eggs/female; 97% for Eqqus caballus with 19 eggs/female; 98% for human blood with 14 eggs/female; 71.3% for Didelphis albiventris with 8.4 eggs/female; 73% for Gallus gallus with 14 eggs/female; 86% for Canis familiaris with 10.3 eggs/female; 81.4% for Galea spixii with 26 eggs/female; 36% for Callithrix jachus with 15 eggs/female; 42.8% for Monodelphis domestica with 0% of oviposition. Female sand flies did not take a blood meal from Felis catus. Sand flies life cycle ranged from 32-40 days, with 21-50 oviposition rates approximately. This study also showed that at 32°C the life cycle had 31 days, at 28° C it had 50 days and at 22°C it increased to 79 days. Adjusting the temperature to 35°C the eggs did not hatch, thus blocking the life cycle. A total of 1540 sand flies were captured, among them, 1.310 were male and 230 were female. Whereas 86% of the sand flies captured were Lu. longipalpis as compared to 10.5% for Lu. evandroi and, 3.2% for L. lenti and 0.3% for Lu whitmani. The ratio between female and male sandfly was approximately 6 males to 1 female. In Nísia Floresta, 50.7% of the collected females took their blood meal from armadillo, 12.8% from human. Among the female sand flies captured in São Gonçalo do Amarante, 80 of them were tested for the Leishmania KDNA infectivity where 5% of them were infected with Leishmania chagasi. Female Lutzomyia spp. showed to have an opportunistic blood meal characteristic. The behavioral parameters seem to have a higher influence in the oviposition when compared to the level of total proteins detected in the host s bloodstream. A higher Lu. longipalpis life cycle viability was observed at 28°C. The increase of temperature dropped the life cycle time, which means that the life cycle is modified by temperature range, source of blood meal and humidity. Lu longipalpis was the most specie found in the inner and peridomiciliar environment. In Nísia Floresta, armadillos were the main source of blood meal for Lutzomyia spp. At São Gonçalo do Amarante, humans were the main source of blood meal due to CDC nets placed inside their houses