Relation of pretreatment sequence diversity in NS5A region of HCV genotype 1 with immune response between pegylated-INF/ribavirin therapy outcomes

Hepatitis C virus (HCV) infection frequently persists despite substantial virus-specific immune responses and the combination of pegylated interferon (INF)-alpha and ribavirin therapy. Major histocompatibility complex class I restricted CD8+ T cells are responsible for the control of viraemia in HCV infection, and several studies suggest protection against viral infection associated with specific HLAs. The reason for low rates of sustained viral response (SVR) in HCV patients remains unknown. Escape mutations in response to cytotoxic T lymphocyte are widely described; however, its influence in the treatment outcome is ill understood. Here, we investigate the differences in CD8 epitopes frequencies from the Los Alamos database between groups of patients that showed distinct response to pegylated alpha-INF with ribavirin therapy and test evidence of natural selection on the virus in those who failed treatment, using five maximum likelihood evolutionary models from PAML package. The group of sustained virological responders showed three epitopes with frequencies higher than Non-responders group, all had statistical support, and we observed evidence of selection pressure in the last group. No escape mutation was observed. Interestingly, the epitope VLSDFKTWL was 100% conserved in SVR group. These results suggest that the response to treatment can be explained by the increase in immune pressure, induced by interferon therapy, and the presence of those epitopes may represent an important factor in determining the outcome of therapy.

Variety matters: adaptive genetic diversity and parasite load in two mouse opossums from the Brazilian Atlantic forest

The adaptive potential of a species to a changing environment and in disease defence is primarily based on genetic variation. Immune genes, such as genes of the major histocompatibility complex (MHC), may thereby be of particular importance. In marsupials, however, there is very little knowledge about natural levels and functional importance of MHC polymorphism, despite their key role in the mammalian evolution. In a previous study, we discovered remarkable differences in the MHC class II diversity between two species of mouse opossums (Gracilinanus microtarsus, Marmosops incanus) from the Brazilian Atlantic forest, which is one of the most endangered hotspots for biodiversity conservation. Since the main forces in generating MHC diversity are assumed to be pathogens, we investigated in this study gastrointestinal parasite burden and functional associations between the individual MHC constitution and parasite load. We tested two contrasting scenarios, which might explain differences in MHC diversity between species. We predicted that a species with low MHC diversity would either be under relaxed selection pressure by low parasite diversity (`Evolutionary equilibrium` scenario), or there was a recent loss in MHC diversity leading to a lack of resistance alleles and increased parasite burden (`Unbalanced situation` scenario). In both species it became apparent that the MHC class II is functionally important in defence against gastrointestinal helminths, which was shown here for the first time in marsupials. On the population level, parasite diversity did not markedly differ between the two host species. However, we did observe considerable differences in the individual parasite load (parasite prevalence and infection intensity): while M. incanus revealed low MHC DAB diversity and high parasite load, G. microtarsus showed a tenfold higher population wide MHC DAB diversity and lower parasite burden. These results support the second scenario of an unbalanced situation.

Molecular characterization of a retrotransposon in the Rhynchosciara americana genome and its association with telomere

Non-LTR retrotransposons, also known as long interspersed nuclear elements (LINEs), are transposable elements that encode a reverse transcriptase and insert into genomic locations via RNA intermediates. The sequence analysis of a cDNA library constructed from mRNA of the salivary glands of R. americana showed the presence of putative class I elements. The cDNA clone with homology to a reverse transcriptase was the starting point for the present study. Genomic phage was isolated and sequenced and the molecular structure of the element was characterized as being a non-LTR retrotransposable element. Southern blot analysis indicated that this transposable element is represented by repeat sequences in the genome of R. americana. Chromosome tips were consistently positive when this element was used as probe in in-situ hybridization. Real-time RT-PCR showed that this retrotransposon is transcribed at different periods of larval development. Most interesting, the silencing of this retrotransposon in R. americana by RNA interference resulted in reduced transcript levels and in accelerated larval development.

The Immunomodulatory Action of Sialostatin L on Dendritic Cells Reveals Its Potential to Interfere with Autoimmunity

Sialostatin L (SialoL) is a secreted cysteine protease inhibitor identified in the salivary glands of the Lyme disease vector Ixodes scapularis. In this study, we reveal the mechanisms of SialoL immunomodulatory actions on the vertebrate host. LPS-induced maturation of dendritic cells from C57BL/6 mice was significantly reduced in the presence of SialoL. Although OVA degradation was not affected by the presence of SialoL in dendritic cell cultures, cathepsin S activity was partially inhibited, leading to an accumulation of a 10-kDa invariant chain intermediate in these cells. As a consequence, in vitro Ag-specific CD4(+) T cell proliferation was inhibited in a time-dependent manner by SialoL, and further studies engaging cathepsin S(-/-) or cathepsin L(-/-) dendritic cells confirmed that the immunomodulatory actions of SialoL are mediated by inhibition of cathepsin S. Moreover, mice treated with SialoL displayed decreased early T cell expansion and recall response upon antigenic stimulation. Finally, SialoL administration during the immunization phase of experimental autoimmune encephalomyelitis in mice significantly prevented disease symptoms, which was associated with impaired IFN-gamma and IL-17 production and specific T cell proliferation. These results illuminate the dual mechanism by which a human disease vector protein modulates vertebrate host immunity and reveals its potential in prevention of an autoimmune disease. The Journal of Immunology, 2009, 182: 7422-7429.

Alveolar macrophages from susceptible mice are more competent than those of resistant mice to control initial Paracoccidioides brasiliensis infection

Alveolar macrophages ( AM) are the first host cells to interact with Paracoccidioides brasiliensis (Pb), a primary human pathogen that causes severe pulmonary infections in Latin America. To better understand innate immunity in pulmonary paracoccidioidomycosis, we decided to study the fungicidal and secretory abilities of AM from resistant (A/J) and susceptible (B10.A) mice to infection. Untreated, IFN-gamma and IL-12 primed AM from B10. A and A/J mice were challenged with P. brasiliensis yeasts and cocultured for 72 h. B10. A macrophages presented an efficient fungicidal ability, were easily activated by both cytokines, produced high levels of nitric oxide ( NO), IL-12, and MCP-1 associated with low amounts of IL-10 and GM-CSF. In contrast, A/J AM showed impaired cytokine activation and fungal killing, secreted high levels of IL- 10 and GM-CSF but low concentrations of NO, IL- 12, and MCP-1. The fungicidal ability of B10. A but not of A/J macrophages was diminished by aminoguanidine treatment, although only the neutralization of TGF-beta restored the fungicidal activity of A/J cells. This pattern of macrophage activation resulted in high expression of MHC class II antigens by A/J cells, while B10. A macrophages expressed elevated levels of CD40. Unexpectedly, our results demonstrated that susceptibility to a fungal pathogen can be associated with an efficient innate immunity, while a deficient innate response can ultimately favor the development of a resistant pattern to infection. Moreover, our data suggest that different pathogen recognition receptors are used by resistant and susceptible hosts to interact with P. brasiliensis yeasts, resulting in divergent antigen presentation, acquired immunity, and disease outcomes.

Preventive and curative glycoside kaempferol treatments attenuate the TH2-driven allergic airway disease

Asthma is a chronic respiratory disease characterized by airway inflammation and airway hyperresponsiveness (AHR). One strategy to treat allergic diseases is the development of new drugs. Flavonoids are compounds derived from plants and are known to have antiallergic, anti-inflammatory, and antioxidant properties. To investigate whether the flavonoid kaempferol glycoside 3-O-[beta-D-glycopiranosil-(1 -> 6)-alpha-L-ramnopiranosil]-7-O-alpha-L-ramnopiranosil-kaempferol (GRRK) would be capable of modulating allergic airway disease (AAD) either as a preventive (GRRK P) or curative (GRRK C) treatment in an experimental model of asthma. At weekly intervals, BALB/c mice were subcutaneously (sc) sensitized twice with ovalbumin (OVA)/alum and challenged twice with OVA administered intranasally. To evaluate any preventive effects GRRK was administered 1 h (hour) before each OVA-sensitization and challenge, while to analyze the curative effects mice were first sensitized with OVA, followed by GRRK given at day 18 through 21. The onset: of AAD was evaluated 24 h after the last OVA challenge. Both treatments resulted in a dose-dependent reduction in total leukocyte and eosinophil counts in the bronchoalveolar lavage fluid (BAL). GRRK also decreased CD4(+), B220(+), MHC class II and CD40 molecule expressions in BAL cells. Histology and lung mechanic showed that GRRK suppressed mucus production and ameliorated the AHR induced by OVA challenge. Furthermore, GRRK impaired Th2 cytokine production (IL-5 and IL-13) and did not induce a Th1 pattern of inflammation. These findings demonstrate that GRRK treatment before or after established allergic lung disease down-regulates key asthmatic features. Therefore. GRRK has a potential clinical use for the treatment of allergic asthma. (C) 2009 Elsevier B.V. All rights reserved.

Automatic contour extraction from 2D neuron images

This work describes a novel methodology for automatic contour extraction from 2D images of 3D neurons (e.g. camera lucida images and other types of 2D microscopy). Most contour-based shape analysis methods cannot be used to characterize such cells because of overlaps between neuronal processes. The proposed framework is specifically aimed at the problem of contour following even in presence of multiple overlaps. First, the input image is preprocessed in order to obtain an 8-connected skeleton with one-pixel-wide branches, as well as a set of critical regions (i.e., bifurcations and crossings). Next, for each subtree, the tracking stage iteratively labels all valid pixel of branches, tip to a critical region, where it determines the suitable direction to proceed. Finally, the labeled skeleton segments are followed in order to yield the parametric contour of the neuronal shape under analysis. The reported system was successfully tested with respect to several images and the results from a set of three neuron images are presented here, each pertaining to a different class, i.e. alpha, delta and epsilon ganglion cells, containing a total of 34 crossings. The algorithms successfully got across all these overlaps. The method has also been found to exhibit robustness even for images with close parallel segments. The proposed method is robust and may be implemented in an efficient manner. The introduction of this approach should pave the way for more systematic application of contour-based shape analysis methods in neuronal morphology. (C) 2008 Elsevier B.V. All rights reserved.

Cathepsin S as a biomarker of low-grade inflammation, insulin resistance, and cardiometabolic disease risk

Cathepsin S is a protease important in major histocompatibility complex (MHC) class II antigen presentation and also in degrading the extracellular matrix. Studies, most of them experimental, have shown that cathepsin S is involved in different pathological conditions such as obesity, inflammation, atherosclerosis, diabetes, and cancer.    The overall hypothesis of this report is that high levels of circulating cathepsin S, is a biomarker that reflects pathology induced by inflammation and obesity. The overall aim of this report was to investigate possible associations between circulating cathepsin S, inflammation, glucometabolic disturbance, and its associated diseases in the community. As cathepsin S appears to be a novel risk marker for several pathological conditions, we also wanted to examine the effect of dietary intervention on circulating cathepsin S concentrations.    This thesis is based on data from three community-based cohorts, the Uppsala longitudinal study of adult men (ULSAM), the prospective investigation of the vasculature in Uppsala seniors (PIVUS), and a post-hoc study from the randomized controlled NORDIET trial.    In the first study, we identified a cross-sectional positive association between serum cathepsin S and two markers of cytokine-mediated inflammation, CRP and IL-6. These associations were similar in non-obese individuals. In longitudinal analyses, higher cathepsin S at baseline was associated with higher CRP and IL-6 levels after six years of follow-up. In the second study, we identified a cross-sectional association between increased serum levels of cathepsin S and reduced insulin sensitivity. These associations were similar in non-obese individuals. No significant association was observed between cathepsin S and insulin secretion. In longitudinal analysis, higher cathepsin S levels were associated with an increased risk of developing diabetes during the six-year follow-up. In the third study, we found that higher serum levels of cathepsin S were associated with increased mortality risk. Moreover, in the ULSAM cohort, serum cathepsin S was independently associated with cause-specific mortality from cardiovascular disease and cancer. In the fourth study, we identified that adherence to an ad libitum healthy Nordic diet for 6 weeks slightly decreased the levels of plasma cathepsin S in normal or marginally overweight individuals, relative to the control group. Changes in circulating cathepsin S concentrations were correlated with changes in body weight, LDL-C, and total cholesterol.    Conclusion: This thesis shows that circulating cathepsin S is a biomarker that independently reflects inflammation, insulin resistance, the risk of developing diabetes, and mortality risk. Furthermore, a Nordic diet moderately reduced cathepsin S levels in normal-weight and overweight men and women. This effect may be partially mediated by diet-induced weight loss and possibly by reduced LDL-C concentrations. 

O ambiente socio-econômico como fator preditivo nas chances de classificação no vestibular e no êxito acadêmico na universidade: (um estudo da clientela da Fundação CESGRANRIO, Rio de Janeiro - 1975)

o presente trabalho teve como objetivo estudar a influência que o ambiente sócio-econômico poderia exercer sob re as chances do candidato no vestibular e sobre o êxito acadêmico do universitário. Para tanto re alizaram-se duas pesquisas: uma com os candidatos do vestibular unificado da Fundação CESGRANRIO. do Rio de Ja neira, de 1975: outra, com uma amostra de universitários classificados p~ 10 mesmo vest i bular. O pri meiro estudo abrangeu 75.348 candidatos e o segundo 1.608 universitários, de sete instituições e nove carreiras diferentes. Uti lizou-se como instrumentos de pesquisa: um I questionário de informação sócio-cultural. o total de pontos obtidos pelos candidatos no vestibular e a méd ia das notas obtidas no curso superior p~ lo estudante, nos deis primeiros semestres de 1975. A anãlise dos resultados permitiu concluir o se guinte: - Tiveram maior chance de classificação no vest i bular os candidatos que: cursaram o clássico; f reqüentaram tanto escola / particu lar como oficial; cursaram o período diurno, obtiveram concei to "E~ ce lente" no 2? grau; não trabalhavam; possuíam pai com nível de ins trução superior e renda mensal familiar acima de 8.000 cruzei ros. - Exercem influência na classificação do vestibu lar os seguintes indicadores do amb iente sócio-econôm ico: renda mensal fa miliar, nível de instrução e profissão do pai. ativ idade ocupac ional do / candidato e período da escola de 2? grau. Obtiveram bom rendimento na universidade os estudantes que: freqüentaram o período diurno; obtiveram bom rendimento no 2? grau; não exerciam atividade ocupac ional: possuiam pais com profissão / liberal e renda familiar alta. - São os segu intes os indicadores do ambiente sac io-econôm ico do univers i tário que têm valor preditivo no êxito acadêmico: período da escola de 2? grau, at ividade ocupacional. profissão do pai. ren dimento escolar no 2~ grau e renda mensal fami l iar. Vale assinalar que esses resultados se referem aos candidatos do vestibular unificado da Fundação CESGRANRIO.

Prevalência de oclusopatias e caracteríticas cefalométricas e dentais de escolares cearense da cidade de Fortaleza

The people of Ceará state are descended from miscegenation between the Portuguese colonizers and the native population, resulting in a different facial pattern from other populations. It is important that this pattern be thoroughly understood, along with its minimum and maximum values so that they can be assessed and respected, allowing professionals who deal with the craniofacial complex to work more efficiently and scientifically. Aim: To characterize the morphological pattern of individuals from Ceará state, whose father and grandfather are also native from Ceará, in the 10-12 year age group, not submitted to previous orthodontic treatment, in order to determine: 1) the prevalence of occlusal pattern; 2) the prevalence of dental anomalies (DA) and, 3) the skeletal and dental cephalometric characteristics of individuals that present with normal occlusion and harmonious facial pattern. Methodology: A list of 10-12 year-olds was obtained from 515 schools containing 162,713 students (Education Secretariat of Ceará State), from which 234 individuals were examined (107 boys and 157 girls). The assessment criteria adopted were: 1) Angle s Classification System to determine occlusal pattern. The occlusal characteristics were measured through overbite, overjet, crowding and interincisal diastema. 2) DA are anomalies of number, shape, size, eruption and structure and, 3) in the group that presented with normal occlusion, we used cephalometric analysis measures proposed by Downs, Steiner, Tweed, Holdaway, Jacobson and McNamara. Results: 1) 25.8% of the schoolchildren had normal occlusion, 47.5% class I malocclusion, 22.3% class II malocclusion and 4.2% class III malocclusion. No statistically significant difference was found between the age group studied and sex. Thirty percent of the individuals had normal overbite, while 36.7% and 19.7% had increased and reduced overbite, respectively. Normal overjet was found in 33.7% of the individuals, increased overjet in 50% and reduced in 16.3%. Dental crowding was observed in 62.5% of the individuals and the presence of interincisal diastema in 14.8%. 2) The prevalence of DA was 56.1%, 6.8% in the number, 10.8% in shape, 4.1% in size, 34.5% in eruption, 26.4% in structure and 17.4% had more than one DA. No association was found between DA and sex, but DA was significantly associated to malocclusion (p<0.05); 3) there was no association between sex or facial type between the measures of nasal-labial angle, position and effective maxillary length, effective mandibular length and the sagittal relationship between the molars, overjet and overbite, position of upper incisors, lower incisors and between the incisors themselves. There was a difference between sex, on the VERT index and in lower anterior facial height, upper incisor inclination and line-H, between facial types for the occlusal plane angles, mandibular plane, facial axis, lower incisor inclination, mandibular position, upper incisor position, lower anterior facial height, ANB and line-H. It was concluded that: 1) the most prevalent occlusal type was class I malocclusion, with no distinction for sex or age group, and the assessment of occlusal characteristics showed that excessive overbite and overjet were the most predominant findings, along with a high occurrence of tooth crowding; 2) a high prevalence of DA was found, particularly eruption anomalies, not influenced by sex but significantly associated to malocclusion and 3) individuals from Ceará are predominantly brachyfacial, exhibiting a number of similarities inherent to their facial pattern, such as a convex profile, retracted jaw, reduced lower third and protruded lower incisors. This study was multidisciplinary, involving researchers from the areas of epidemiology, radiology and dentistry, thereby meeting the multidisciplinarity requirements of the Postgraduate Program in Health Sciences

O tratamento ortodôntico em uma perspectiva a longo prazo: avaliação da estabilidade oclusal e do grau de satisfação dos pacientes

This study aimed to evaluate factors associated to orthodontic treatment stability and patient satisfaction in the long-term. A total of 209 patients (88 class I and 121 class II) treated with straight wire fixed appliance were selected at least 5 years post treatment. Six hundred twenty seven dental casts were examined with the PAR Index at pretreatment (T1), end of treatment (T2), and at long-term follow up (T3, mean 8.5 years post treatment). At T3, a Dental Impact on Daily Living questionnaire was used to assess patient satisfaction with the dentition in the long-term. Friedman test and multiple regression analysis were used to evaluate changes among the time points and factors associated with stability and patient satisfaction. Predictive factors used to exam the occlusion were: PAR Index at T1 and T2, age at T1, the amount of time without retainer, length of Hawley retainer wear, length of follow-up, sex, extraction and third molar status. To assess patient satisfaction were considered: changes produced by the orthodontic treatment (PAR T2-T1), post treatment stability (PAR T3), age at the start of treatment (T1), length of treatment (T2-T1), gender, and extraction. Orthodontic treatment produced a significant improvement of 94.2% in the PAR Index (T2-T1), but this change was not associated with the level of satisfaction when the patient was questioned at T3. No significant change was observed between T2 and T3. However, when the sample was divided according to the level of finalization (PAR T2), it was observed that well-finished patients experienced some deterioration (P<.001), whereas the less well-finished ones showed some improvement (P<.05). Even with the deterioration, the well-finished patients still had a better PAR Index at T3 compared to the less well-finished ones (PAR T2- T3). Regression analysis showed that PAR Index at T1 and T2, age at T1, and length of retainer wear had a slight association with occlusal stability (R2 = 0.27). Patient satisfaction was significantly associated only with PAR Index at T3 (r2=0.125, P<.0001). We can conclude that, even thought orthodontic treatment is quite stable, not so well-finished treatments tend to show some improvement and well-finished ones deteriorate some in the long-term. Despite of that, well-finished patients still have better occlusal characteristics. Patient satisfaction is not related to the result of orthodontic treatment; nevertheless, there is a slight association with dentition in the long-term

Avaliação de indicadores de impactos ambientais no perímetro irrigado Cruzeta-Região semi-árida do RN

Environmental impacts are defined as the processes of social and ecological changes caused by nuisance ambient. In agriculture are diverse, and the intensive use of land and inappropriate agricultural practices causes negative effect on the environment besides affecting crop productivity and quality of life of man. So this study was objective to analyze parameters indicators of environmental impacts in the Cruzeta the Discrict Irrigation. During the period July 2007 to March 2008 samples were collected in lot 01 and 02 of the irrigation district. The monitoring was conducted in four sampling points, three (3) located in the irrigation channels and 1 (one) located in a cacimbão. Were monitored pH, CE, STD, SS, NO3-, OD, DT, PST, RAS, CT, Ca+2, Mg+2, Na+ and K+ and heavy metals. Analysis was performed, soil fertility, determining the infiltration rate, moisture content of soil and flow measurement. The results showed that the parameters considered most effective in evaluating the indicators of environmental impacts were: Suspended solid, turbidity, dissolved oxygen and Coliform termotelerantes. The concentrations of nitrate were below the limit established by CONAMA (2005). High levels of STD were found in the point P4. The risk of salinity in cacimbão Lot 02, the water was classified as Class II, or medium risk of salinity in the remaining points was classified as Class I. The iron and aluminum were found high concentrations in four sampling points independent of the dry or rainy. On the ground, found high levels of phosphorus in both samples collected in the batch 01, as the lot 02. The pH levels found in samples of soil, the soil can be classified as neutral and moderate alkalinity. The high infiltration rate obtained in the tests performed in lots 01 and 02 indicated the high permeability of soil at these points

Expression of nonclassical molecule human leukocyte antigen-G in oral lesions

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Upregulation of soluble and membrane-bound human leukocyte antigen G expression is primarily observed in the milder histopathological stages of chronic hepatitis C virus infection

Chronic hepatitis C virus (HCV) infection is a worldwide health problem that may evolve to cirrhosis and hepatocellular carcinoma. Incompletely understood immune system mechanisms have been associated with impaired viral clearance. The nonclassical class I human leukocyte antigen G (HLA-G) molecule may downregulate immune system cell functions exhibiting well-recognized tolerogenic properties. HCV genotype was analyzed in chronic HCV-infected patients. Because HLA-G expression may be induced by certain viruses, we evaluated the presence of HLA-G in the liver microenvironment obtained from 89 biopsies of patients harboring chronic HCV infection and stratified according to clinical and histopathological features. Overall, data indicated that HCV genotype 1 was predominant, especially subgenotype 1a, with a prevalence of 87%. HLA-G expression was observed in 45(51%) liver specimens, and it was more frequent in milder stages of chronic hepatitis (67.4%) than in moderate (27.8%; p = 0.009) and severe (36.0%; p = 0.021) stages of the disease. Altogether, these results suggest that the expression of HLA-G in the context of HCV is a complex process modulated by many factors, which may contribute to an immunologic environment favoring viral persistence. However, because the milder forms predominantly expressed HLA-G, a protective role of this molecule may not be excluded. (C) 2012 American Society for Histocompatibility and Immunogenetics. Published by Elsevier B.V. All rights reserved.

Análise comparativa na influência da técnica de moldagem funcional em arcos parcialmente edêntulos sobre os pontos de contato oclusais, a integridade da fibromucosa e a extensão da base da prótese

There is a lack of clinical studies evaluating techniques of functional impression for partially edentulous arches. The aim of this double-blind non-randomized controlled clinical trial was to compare the efficacy of altered cast impression (ACI) and direct functional impression (DFI) techniques. The efficacy was evaluated regarding the number of occlusal units on denture teeth, mucosa integrity at 24-hour follow-up and denture base extension. The sample included 51 patients (female and male) with mean age of 58.96 years treated at Dental Department of UFRN. The patients, exhibiting edentulous maxilla and mandibular Kennedy class I, were divided into two groups (group ACI, n=29; group DFI, n=22). Clinical evaluation was based on the number of occlusal units on natural and/or artificial teeth, mucosa integrity at 24-hour follow-up, and denture base extension. Statistical analysis was conducted using the software SPSS 17.0® (SPSS Inc., Chicago, Illinois). Student T-test was used to reveal association between number of occlusal units and impression technique while chi-square test showed association between mucosa integrity and impression technique. Fischer s exact test was applied for association between denture base extension and impression technique at 95% level of significance. No significant difference was observed between the groups regarding number of occlusal units, mucosa integrity and denture base extension. The altered cast technique did not provide significant improvement in comparison to the direct technique when the number of occlusal units, mucosa integrity and denture base extension