62 resultados para Resposta fisiológica
Resumo:
The Chromobacterium violaceum is a β-proteobacterium Gram-negative widely found in tropical and subtropical regions, whose genome was sequenced in 2003 showing great metabolic versatility and biotechnological and pharmaceutical potential. Given the large number of ORFs related to iron metabolism described in the genome of C. violaceum, the importance of this metal for various biological processes and due to lack of data about the consequences of excess of iron in free-living organisms, it is important to study the response mechanism of this bacterium in a culture filled with iron. Previous work showed that C. violaceum is resistant to high concentrations of this metal, but has not yet been described the mechanism which is used to this survival. Thus, to elucidate the response of C. violaceum cultured in high concentrations of iron and expecting to obtain candidate genes for use in bioremediation processes, this study used a shotgun proteomics approach and systems biology to assess the response of C. violaceum grown in the presence and absence of 9 mM of iron. The analysis identified 531 proteins, being 71 exclusively expressed by the bacteria grown in the presence of the metal and 100 just in the control condition. The increase in expression of proteins related to the TCA cycle possibly represents a metabolic reprogramming of the bacteria caused by high concentration of iron in the medium. Moreover, we observed an increase in the activity assay of superoxide dismutase and catalase as well as in Total Antioxidant Activity assay, suggesting that the metal is inducing oxidative stress in C. violaceum that increases the levels of violacein and antioxidant enzymes to better adapt to the emerging conditions. Are also part of the adaptive response changes in expression of proteins related to transport, including iron, as well as an increased expression of proteins related to chemotaxis response, which would lead the bacteria to change the direction of its movement away from the metal. Systems Biology results, also suggest a metabolic reprogramming with mechanisms coordinated by bottleneck proteins involved in transcription (GreA), energy metabolism (Rpe and TpiA) and methylation (AhcY)
Resumo:
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.
Resumo:
Despite advances in antibiotic therapy, bacterial meningitis (BM) remains with high mortality and morbidity rates in worldwide. One important mechanism associated to sequels during disease is the intense inflammatory response which promotes an oxidative burst and release of reactive oxygen species, consequently leading to cell death. Activation of DNA repair enzymes during oxidative stress has been demonstrated in several neurological disorders. APE1/Ref-1 is a multifunctional protein involved in DNA repair and plays a redox function on transcription factors such as NFkB and AP-1.The aim of this study was assess the role of APE1/Ref-1 on inflammatory response and the possibility of its modulation to reduce the sequels of the disease. Firstly it was performed an assay to measure cytokine in cerebrospinal fluid of patients with BM due to Streptococcus pneumoniae and Neisseriae meningitides. Further, a cellular model of inflammation was used to observe the effect of the inhibition of the endonuclease and redox activity of APE1/Ref-1 on cytokine levels. Additionally, APE1/Ref-1 expression in cortex and hippocampus of rat with MB after vitamin B6 treatment was evaluated. Altogether, results showed a similar profile of cytokines in the cerebrospinal fluid of patients from both pathogens, although IFNy showed higher expression in patients with BM caused by S. pneumoniae. On the other hand, inhibitors of APE1/Ref-1 reduced cytokine levels, mainly TNF-α. Reduction of oxidative stress markers was also observed after introduction of inhibitors in the LPS-stimulated cell. In the animal model, BM increased the expression of the protein APE1/Ref-1, while vitamin B6 promoted reduction. Thereby, this data rise important factors to be considered in pathogenesis of BM, e.g., IFNy can be used as prognostic factor during corticosteroid therapy, APE1/Ref-1 can be an important target to modulate the level of inflammation and VIII oxidative stress, and vitamin B6 seems modulates several proteins related to cell death. So, this study highlights a new understanding on the role of APE1/Ref-1 on the inflammation and the oxidative stress during inflammation condition
Resumo:
In vitro and in animal models, APE1, OGG1, and PARP-1 have been proposed as being involved with inflammatory response. In this work, we have investigated if the SNPs APE1 Asn148Glu, OGG1 Ser326Cys, and PARP-1 Val762Ala are associated to meningitis and also developed a system to enable the functional analysis of polymorphic proteins. Patients with bacterial meningitis (BM), aseptic meningitis (AM) and controls (non-infected) genotypes were investigated by PIRA-PCR or PCR-RFLP. DNA damages were detected in genomic DNA by Fpg treatment. IgG and IgA were measured from plasma and the cytokines and chemokines were measured from cerebrospinal fluid samples using Bio-Plex assays. The levels of NF-κB and c-Jun were measured in CSF by dot blot assays. A significant (P<0.05) increase in the frequency of APE1 148Glu allele in BM and AM patients was observed. A significant increase in the genotypes Asn/Asn in control group and Asn/Glu in BM group was also found. For the SNP OGG1 Ser326Cys, the genotype Cys/Cys was more frequent (P<0.05) in BM group. The frequency of PARP-1 Val/Val genotype was higher in control group (P<0.05). The occurrence of combined SNPs increased significantly in BM patients, indicating that these SNPs may be associated to the disease. Increasing in sensitive sites to Fpg was observed in carriers of APE1 148Glu allele or OGG1 326Cys allele, suggesting that SNPs affect DNA repair activity. Alterations in IgG production were observed in the presence of SNPs APE1Asn148Glu, OGG1Ser326Cys or PARP-1Val762Ala. Reductions in the levels ofIL-6, IL-1Ra, MCP-1/CCL2and IL-8/CXCL8 were observed in the presence of APE1148Glu allele in BM patients, however no differences were observed in the levels of NF-κB and c-Jun considering genotypes and analyzed groups. Using APE1 as model, a system to enable the analysis of cellular effects and functional characterization of polymorphic proteins was developed using strategies of cloning APE1 cDNA in pIRES2-EGFP vector, cellular transfection of the construction obtained, siRNA for endogenous APE1 and cellular cultures genotyping. In conclusion, we obtained evidences of an effect of SNPs in DNA repair genes on the regulation of immune response. This is a pioneering work in the field that shows association of BER variant enzymes with an infectious disease in human patients, suggesting that the SNPs analyzed may affect immune response and damage by oxidative stress level during brain infection. Considering these data, new approaches of functional characterization must be developed to better analysis and interactions of polymorphic proteins in response to this context
Resumo:
The genital HPV infection is very common between men and women worldwide, affecting particularly young women, constituting a serious public health problem in less developed regions, favored by the poor living conditions of population. The cytology and colposcopy have notorious importance in the diagnosis of precursor lesions of cervical cancer and therefore its prevention. However, even with such diagnostic tools, the number of women who develop cervical cancer is still high. This study aims to assess the prevalence of genital tract infection by HPV in pregnant and nonpregnant women, evaluating the profile of the immune response presented by the women of these two groups in order to establish correlations among profile of immune response, presence of virus and occurrence of lesions of the uterine cervix. We analyzed specimens obtained from the cervix of 221 patients, 91 pregnant and 130 non-pregnant, aged 14-72 years. The women were subjected to colposcopic and cytologic evaluation detect possible changes in the cervix and then samples were collected in order to perform HPV detection by PCR and real-time PCR for detection of mRNA of pro-inflammatory and anti-inflammatory cytokines. In the present study, the overall prevalence of HPV genital infection was 28.1%; of which 31.9% were pregnant patients and 25.4% in non-pregnant women. Young women under 30 years and those with low educational level education showed a higher risk of HPV infection. Colposcopy showed better correlation with detection of HPV DNA by PCR, when compared to cytology. Generally, HPV infected patients, pregnant or not, exhibited reduced mRNA expression of both pro-inflammatory (IFN-γ, TNF-α) and anti-inflammatory (IL -10) cytokines, when compared to patients not infected by HPV. Nonpregnant patients infected presented increase mRNA expression of IL-17 in patients without injury, whereas those with lesion showed higher mRNA expression of TGF-β. Pregnant women without injury infected exhibited increased mRNA expression of TGF-β. There was no difference in HPV prevalence between pregnant and nonpregnant women. There was a reduction of pro-inflammatory cytokines, except IL-17, in all women infected by HPV. Moreover, we observed an increase of TGF-β in HPV-infected women who are pregnant or not. The results suggest that, in women in this study, HPV infection promoted changes in the profile of cytokines necessary for activation of effective immune response, possibly favoring viral persistence
Resumo:
Leishmania chagasi infection presents a wide spectrum of clinical outcomes, ranging from asymptomatic self resolving infection to disease, visceral leishmaniasis (VL). The exact mechanisms that lead the evolution of infection to disease are not understood. It is believed that malnutrition is a risk factor associated with VL development, although there are few human studies in the area. We aimed to assess the nutritional factors associated with the response to L. chagasi infection in Rio Grande do Norte. The study was conducted from December 2006 to January 2008. 149 children were assessed: 20 active VL cases, 33 children with VL history, 40 DTH+ asymptomatic children and 56 DTH-. Nutritional status was assessed using z scores for Weight/Age, Weight/Height, Height/Age, Body Mass Index (BMI), and mid-upper arm circumference/height (MUAC/height). Vitamin A status was determined by serum retinol concentrations and the modified-relative-dose-esponse test (MRDR). Breastfeeding time and birth weight were also evaluated. VL children presented compromised nutritional status when compared to the other groups using BMI and MUAC/age, with means -1,53 ± 1,10 and -1,48 ± 1,28 z scores, respectively (ANOVA, p < 0,05). VL children also showed lower vitamin A levels: 43% presented serum retinol < 20 µg/dL and 15% MRDR > 0,060. Birth weight was inverserly associated with the risk to belong the VL group (β = -0,00; OR = 0,84; 95% CI 0,73 - 0,99; p = 0,047), whereas more breastfeeding time was directly associated with the risk to belong to the DTH+ group (β = 0,02; OD = 1,16; 95% CI 1,01 - 1,33; p = 0,036). The nutritional variables evaluated were associated with the response to the L. chagasi infection, with malnutrition and compromised vitamin A status as markers of children who present with VL. Higher birth weight was associated with protection to disease, and higher breastfeeding time was associated with increased likelihood of an asymptomatic infection. The results show that modifiable nutritional aspects in the study population are associated with the response to the L. chagasi infection
Resumo:
analisar o efeito do treinamento aeróbio periodizado sobre a aptidão cardiorrespiratória e respostas afetivas em mulheres com síndrome dos ovários policísticos (SOP) e II) investigar se o exercício aeróbio realizado na zona de prazer para essa população atende a recomendação do American College of Sports Medicine (ACSM) no que se refere à intensidade para melhoria da saúde. Metodologia: foram incluídas mulheres na faixa etária entre 18 e 34 anos, com diagnóstico de SOP de acordo com o Consenso de Rotterdam. Para o subestudo I, oito pacientes sedentárias participaram de 16 semanas de treinamento aeróbio com incrementos mensais de intensidade: fase 1 = 60-70% da frequência cardíaca máxima (FCmax); fase 2 = 70-75% da FCmax; fase 3 = 75-80% da FCmax; fase 4 = 80-85% da FCmax. A intervenção foi realizada três vezes por semana, 40 minutos por sessão. Em todas as sessões foram registradas as respostas afetivas (Feeling Scale -5/+5) e a percepção subjetiva do esforço (escala de Borg CR 6-20). Antes e após a intervenção, as voluntárias realizaram teste ergoespirométrico. Para o subestudo II, 11 pacientes realizaram duas sessões de exercício aeróbio na zona de prazer, sendo registrados parâmetros relativos à demanda física através de receptor de GPS (Global Positioning System) de pulso com cardiofrequencímetro acoplado. As pacientes foram instruídas a realizar 40 minutos de exercício guiadas pelas âncoras verbais bom e muito bom (+3 e +5 na Feeling Scale). Resultados: no subestudo I, após 16 semanas de treinamento, houve aumento da aptidão cardiorrespiratória máxima (17,3%) e submáxima (21,5%). As respostas afetivas variaram entre bom (+3,1 ± 0,8) e razoavelmente bom xi (1,0 ± 0,9) e a percepção subjetiva do esforço entre muito leve a leve (10,2 ± 0,7) e um pouco difícil (12,7 ± 0,6) durante a intervenção. No subestudo II, as pacientes exercitaram-se a ~72,5 ± 6% da FC máxima, ~78,5 ± 6% da FC no limiar anaeróbio e passaram > 95% do tempo em intensidade moderada (~82%) e vigorosa (~16%) durante as sessões experimentais. Em média, as voluntárias reportaram as sessões como fácil (percepção subjetiva do esforço da sessão ~2,2 ± 0,7). Conclusões: o programa de treinamento aeróbio periodizado aumentou a aptidão cardiorrespiratória das pacientes analisadas e foi percebido como uma intervenção prazerosa. Adicionalmente, exercício aeróbio realizado de forma prazerosa atende a recomendação do ACSM no que se refere à intensidade para melhoria da saúde
Determinantes envolvidos na resposta imune celular humana à infecção por Leishmania infantum chagasi
Resumo:
Visceral leishmaniasis (VL) is a disease caused by protozoa of the Leishmania donovani complex, whose infection has clinical spectrum ranging from asymptomatic infection to active disease characterized by fever, cachexia, hepatosplenomegaly, and immunosuppression. The healing or protective immunity require an antigen-specific type 1. The Montenegro skin test (DTH) has been interpreted as a marker of protective immunity. However, there is no known correlation between the DTH response to type 1 and DTH and immunity of type 1 are maintained in the long term. Thus, a longitudinal study of 8 years, nested in a cohort family held in Brazil, documented the status of DTH and cytokine production by peripheral blood mononuclear cells in response to antigen-specific stimulation. This study was the interdisciplinary approach of physicians, biochemists, nutritionists, veterinary medicine, biology and statistics. The results show that 46.2% of subjects were analyzed DTH positive at baseline. The prevalence of positive and DTH induration size increased with age (p = 0.0021). 15.7% of individuals positive DTH "retro-converted" the negative and 50.4% (64) of individuals negative DTH became positive. The size of DTH induration was correlated significantly with the antigen-induced production of IFN-γ (r = 0.6186, p = 0.0001). IL-6 was secreted at higher levels in peripheral blood mononuclear cells of individuals who "retro-converted" DTH positive to negative than individuals who remained stable DTH status (p = 0.005). Thus, IFN-γ produced by peripheral blood mononuclear cells, may be a surrogate marker for protective immunity instead of the DTH response. In addition, differences in innate immune response may determine whether individuals maintain or eliminate the infection by Leishmania infantum chagasi in asymptomatic patients
Resumo:
This laboratory study involves the participation of a group with professionals from different areas that had contributed to the construction of a multidisciplinary knowledge, about biological response of titanium surfaces modified through thermochemical treatment by plasma. Thus, the crystalline phase was previously characterized in relation to the topography, roughness, molhability and nitrogen concentration in the samples surface. It s indispensable that materials implanted can influence in a good cellular response as well as promotes a bacteria action. Surfaces modified by plasma were exposed to different cultures such as: cellular (human osteoblastic) and bacteria (Staphylococcus epidermidis ATCC35984 and Pseudomonas aeruginosa ATCC 27853) in order to evaluate the biological response. It was evaluated the adhesion, proliferation, morphology and cellular preference of human ostheoblastic cells (HOST), as well as the formation of a biofilm and bacteria proliferation. It was still analyzed the bacteria selectivity ability in relation to the surfaces. The software Image Pro Plus was used to the counting of cells and bacteria adhered to the surface of disks. The results were submitted to the variance analysis (ANOVA), and then, by the Kruskal-Wallis test, using GraphPad Instat ® software, version 3.5 to Windows. The nitrided samples in spite of show a higher roughness and molhability showed a smaller bacteria growing and higher cellular proliferation, when compared to non treated samples, indicating that the treated material present a high efficiency to biomedical implants
Resumo:
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
Resumo:
The food chain theory predict that presence of omnivory prevent the trophic cascade and could be a strong stabilizing factor over resource and consumer community dynamics, and that the nutrient enrichment destabilize populations dynamics. Most of the freshwater tropical reservoirs are eutrophic, and strategies that seek improve the water quality through the control of phytoplankton biomass and nutrient input, become essential for the improvement and preservation of water quality. The aim of this study was test the zooplanktivory (when larvae) and omnivory (when young and adult) effects of Nile Tilapia over the structure and dynamics of plankton communities, in addition or absence of nutrients enrichment. For this, one field experiment was performed with a factorial design 2x3 resulting in six treatments: control, without fish and nutrient (C); with omnivorous fish (O); with zooplanktivorous fish (Z); without fish and with enrichment of nutrients (NP); with omnivorous fish and nutrients (ONP); and, with zooplanktivorous fish and nutrients (ZNP). The two planktivory types reduced the zooplankton biomass and increased the phytoplankton biomass, but the omnivory of filter-feeding fish attenuated the trophic cascade magnitude. The fertilization by nutrients increases the nutrient concentrations in water and the phytoplankton biomass, but the effect on zooplankton is dependent of the trophic structure. In a general way, the effects of the fish and nutrient addition were addictive, but significant interactions among those factors were observed in the answer of some zooplankton groups. The effects of omnivorous fish over the temporal variability of phytoplankton and zooplankton biomass were very variable, the increase or reduce in variability of the plankton depending of the level of nutrients and of the analyzed variable. With base in this study, we conclude that the planktivory type exercised by the fish and the concentrations of nutrients in the water affects the force of pelagic trophic cascades and probably the success of biomanipulation programs for the handling of water quality in lakes and tropical reservoirs
Resumo:
The complex human behavior related to exercise involves cognitive, physical and emotional processing. The recent theories about exercise s intensity regulation have highlighted the role played by psychophysics aspects in controlling exercise s intensity. In this regard, recent evidences have shown that there is variability in human capacity in perceiving interoceptives clues. Thus, subjects more sensitive show higher physiological arousal to physical and/or emotional stress, and sensations with higher intensity. In fact, studies have evidenced that interoceptive feedback modifies behavior in exercise with free load. However, exercise recommendations are based in a constant load standard. Therefore, we aimed to analyze the influence of interoceptive sensibility on psychophysics responses during dynamic exercise performed with constant load. Twenty-four adult males were allocated into two groups accordingly with their interoceptive sensibility: high sensibility (n=11) and low sensibility (13). They underwent to an incremental test (IT) and then randomly to two sections of moderate and severe exercise intensity for 20 minutes. Heart rate (HR), rating of perceived exertion (RPE), affective feelings (AF), alert state (AS), and percentage of associative thoughts were collect during exercise. A two-way ANOVA with repeated measures was used to assess differences between psychophysics responses. There were differences between group in RPE, AF, and AS in moderate intensity. There was no difference in any measure in severe intensity. We conclude that subjects with high interoceptive sensibility feel dynamic moderate exercise more intense than the subjecs with low interoceptive sensibility
Resumo:
This work aims to show how the application of frequency selective surfaces (FSS) in planar antenna arrays become an alternative to obtain desired radiation characteristics from changes in radiation parameters of the arrays, such as bandwidth, gain and directivity. In addition to analyzing these parameters is also made a study of the mutual coupling between the elements of the array. To accomplish this study, were designed a microstrip antenna array with two patch elements, fed by a network feed. Another change made in the array was the use of the truncated ground plane, with the objective of increasing the bandwidth and miniaturize the elements of the array. In order to study the behavior of frequency selective surfaces applied in antenna arrays, three different layouts were proposed. The first layout uses the FSS as a superstrate (above the array). The second layout uses the FSS as reflector element (below the array). The third layout is placed between two FSS. Numerical and experimental results for each of the proposed configurations are presented in order to validate the research
Resumo:
Composites based on PEEK + PTFE + CARBON FIBER + Graphite (G_CFRP) has increased application in the top industries, as Aerospace, Aeronautical, Petroleum, Biomedical, Mechanical and Electronics Engineering challenges. A commercially available G_CFRP was warmed up to three different levels of thermal energy to identify the main damage mechanisms and some evidences for their intrinsic transitions. An experimental test rig for systematize a heat flux was developed in this dissertation, based on the Joule Effect. It was built using an isothermal container, an internal heat source and a real-time measurement system for test a sample by time. A standard conical-cylindrical tip was inserted into a soldering iron, commercially available and identified by three different levels of nominal electrical power, 40W (manufacturer A), 40W (manufacturer B), 100W and 150W, selected after screening tests: these power levels for the heat source, after one hour of heating and one hour of cooling in situ, carried out three different zones of degradation in the composite surface. The bench was instrumented with twelve thermocouples, a wattmeter and a video camera. The twelve specimens tested suffered different degradation mechanisms, analyzed by DSC (Differential Scanning Calorimetry) and TG (Thermogravimetry) techniques, Scanning Electron Microscopy (SEM) and Energy-Dispersive X-Rays (EDX) Analysis. Before and after each testing, it was measured the hardness of the sample by HRM (Hardness Rockwell M). Excellent correlations (R2=1) were obtained in the plots of the evaporated area after one hour of heating and one hour of cooling in situ versus (1) the respective power of heat source and (2) the central temperature of the sample. However, as resulting of the differential degradation of G_CFRP and their anisotropy, confirmed by their variable thermal properties, viscoelastic and plastic properties, there were both linear and non-linear behaviour between the temperature field and Rockwell M hardness measured in the radial and circumferential directions of the samples. Some morphological features of the damaged zones are presented and discussed, as, for example, the crazing and skeletonization mechanism of G_CFRP
Resumo:
Ensure the integrity of the pipeline network is an extremely important factor in the oil and gas industry. The engineering of pipelines uses sophisticated robotic inspection tools in-line known as instrumented pigs. Several relevant factors difficult the inspection of pipelines, especially in offshore field which uses pipelines with multi-diameters, radii of curvature accentuated, wall thickness of the pipe above the conventional, multi-phase flow and so on. Within this context, appeared a new instrumented Pig, called Feeler PIG, for detection and sizing of thickness loss in pipelines with internal damage. This tool was developed to overcome several limitations that other conventional instrumented pigs have during the inspection. Several factors influence the measurement errors of the pig affecting the reliability of the results. This work shows different operating conditions and provides a test rig for feeler sensors of an inspection pig under different dynamic loads. The results of measurements of the damage type of shoulder and holes in a cyclic flat surface are evaluated, as well as a mathematical model for the sensor response and their errors from the actual behavior